RU2788092C2 - Molecules of antibodies to pd-1 and their use - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: group of inventions is described, including the use of a molecule of an antibody against PD-1 in the treatment of cancer in a subject (options), a method for the treatment of cancer in a subject (options), the use of a pharmaceutical composition including a molecule of an antibody against PD-1, at a dose from about 300 mg to 400 mg once every three weeks or once every four weeks, for the treatment of cancer. In one of options of implementation, a molecule of an antibody against PD-1 is used in the treatment of cancer at a dose from about 300 mg to 400 mg once every three weeks or once every four weeks.

EFFECT: invention expands the arsenal of methods for the treatment of cancer, using a molecule of an antibody against PD-1.

103 cl, 35 dwg, 13 tbl, 7 ex

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority of U.S. Provisional Application 62/269,044 filed December 17, 2015, U.S. Provisional Application 62/331,371 filed May 3, 2016, U.S. Provisional Application 62/344,784 filed June 2, 2016, Provisional Application U.S. Patent Application 62/347,331 filed June 8, 2016, U.S. Provisional Application 62/359,781 filed July 8, 2016, U.S. Provisional Application 62/381,384 filed August 30, 2016, U.S. Provisional Application 62 /400,787, filed September 28, 2016, U.S. Provisional Application 62/414,128, filed October 28, 2016, and U.S. Provisional Application 62/431,846, filed December 9, 2016. The contents of the above applications are hereby incorporated by reference in their entirety.

SEQUENCE LIST

This application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The specified ASCII copy, created on December 14, 2016, is labeled C2160-7015WO_SL.txt and has a size of 280741 bytes.

BACKGROUND OF THE INVENTION

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, AJ et al. (2007) Adv. Immunol. 90:297-339 ). First, an antigen that has been exposed on the surface of antigen-presenting cells (APCs) is presented to an antigen-specific naive CD4 ⁺ T cell. This presentation signals through the T cell receptor (TCR), which directs the T cell to initiate an immune response specific for the presented antigen. Second, various co-stimulatory and inhibitory signals mediated by interactions between APC and certain T-cell surface molecules cause T-cell activation and proliferation and, ultimately, their inhibition.

The immune system is tightly controlled by a network of costimulatory and coinhibitory ligands and receptors. These molecules provide a second signal for T cell activation and provide a balanced network of positive and negative signals to maximize immune responses against infection and thereby limit autoimmunity (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 binding between APC ligands B7.1 (CD80) and B7.2 (CD86) and CD28 and CTLA-4 receptors on the CD4 ⁺ T lymphocyte (Sharpe, AH et al. (2002) Nature Rev. Immunol. 2: 116-126 Lindley, PS 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, RJ 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. al (1996) Immunity 4:535-543).

Other CD28 receptor ligands 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 ligand 1 (PD-L1; B7-H1), programmed death ligand 2 (PD-L2; B7-DC), B7-H3, B7-H-4 and B7-H6 (Collins, M. et al. (2005) Genome Biol. 6:223.1-223.7).

Programmed death protein 1 (PD-1) is an inhibitory protein that is a member of the large 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. PD-1 presumably exists as a monomer because it does not contain the unpaired cysteine residue found in other members of the CD28 family. PD-1 is expressed on activated B cells, T cells, and monocytes.

The PD-1 gene encodes a 55 kDa type I transmembrane protein (Agata et al. (1996) Int Immunol. 8:765-72). Although PD-1 is structurally similar to CTLA-4, it does not contain the MYPPY motif (SEQ ID NO: 236) which is important for the binding of B7-1 and B7-2. Two PD-1 ligands, PD-L1 (B7-H1) and PD-L2 (B7-DC), have been identified and shown to decrease T cell activation after binding to PD-1 (Freeman et al. (2000 ) J. Exp. Med. 192:1027-34; Carter et al. (2002) Eur. J. Immunol. 32:634-43). PD-L1 and PD-L2 are B7 homologues that bind to PD-1 but do not bind to other members of the CD28 family. PD-L1 is prevalent in various human cancers (Dong et al. (2002) Nat. Med. 8:787-9).

PD-1 is known to be an immunoinhibitory protein that negatively regulates TCR signals (Ishida, Y. et al. (1992) EMBO J. 11:3887-3895; Blank, C. et al. (Epub 2006 Dec. 29) Immunol. Immunother 56(5):739-745). The interaction between PD-1 and PD-L1 may act as an immune checkpoint, which may result, for example, in a decrease in tumor-infiltrating lymphocytes, a decrease in T-cell receptor-mediated proliferation, and/or immune escape of cancer cells (Dong et al. (2003) J Mol Med 81:281-7 Blank et al (2005) Cancer Immunol Immunother 54:307-314 Konishi et al (2004) Clin Cancer Res 10:5094-100 Immune suppression can be reversed by inhibiting the local interaction of PD-1 with PD-L1 or PD-L2; the effect is additive when the interaction of PD-1 with PD-L2 is also blocked (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA 99:12293-7; Brown et al. (2003) J. Immunol 170:1257-66).

Given the importance of immune checkpoint pathways in regulating the immune response, there is a need to develop new combination therapies that modulate the activity of immunoinhibitory proteins such as PD-1, resulting in activation of the immune system. Such agents may be used, for example, in cancer immunotherapy and in the treatment of other conditions such as chronic infection.

SUMMARY OF THE INVENTION

The present application discloses, at least in part, antibody molecules (eg, humanized antibody molecules) that bind to programmed death 1 (PD-1) protein with high affinity and specificity. Also provided are nucleic acid molecules encoding antibody molecules, expression vectors, host cells, and methods for producing antibody molecules. Pharmaceutical compositions and dosage forms including antibody molecules are also provided. Anti-PD-1 antibody molecules disclosed herein may be used (alone or in combination with other agents or treatments) for the treatment, prevention and/or diagnosis of disorders such as cancer disorders (e.g. solid tumors and soft tissue tumors) , as well as infectious diseases (for example, chronic infectious disorders or sepsis). Thus, the present application discloses compositions and methods for detecting PD-1, as well as methods for treating various disorders, including cancer and/or infectious diseases, using anti-PD-1 antibody molecules. In some embodiments, the anti-PD-1 antibody molecule is administered or administered at a constant or fixed dose.

Thus, in one aspect of the invention, there is provided a method of treating (eg, inhibiting, reducing, mitigating, or preventing) a disorder, eg, a hyperproliferative condition or disorder (eg, cancer) in a subject. The method includes administering to a subject an anti-PD-1 antibody molecule, such as an anti-PD-1 antibody molecule described herein, at a dose of about 300 mg to 400 mg once every three weeks or once every four weeks. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every three weeks. In other embodiments, the anti-PD-1 antibody molecule is administered at a dose of approximately 400 mg once every four weeks. In some embodiments, the disorder is cancer, such as the cancer described herein. In some embodiments, the cancer is skin cancer, such as Merkel cell cancer or melanoma. In one embodiment, the cancer is Merkel cell cancer. In other embodiments, the cancer is melanoma. In other embodiments, the cancer is breast cancer, such as triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In other embodiments, the cancer is kidney cancer, eg, renal cell carcinoma (eg, clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC)). In other embodiments, the cancer is thyroid cancer, such as anaplastic thyroid carcinoma (ATC). In other embodiments, the cancer is a neuroendocrine tumor (NET), such as an atypical pulmonary carcinoid tumor or a NET in the pancreas, gastrointestinal (GI) tract, or lung. In some embodiments, the cancer is lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is colon and rectal cancer (CRC) (eg, highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC)). In some embodiments, the cancer is a leukemia (eg, acute myeloid leukemia (AML), eg, relapsed or refractory AML or de novo AML). In some embodiments, the cancer is a myelodysplastic syndrome (MDS) (eg, high risk MDS).

In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks or about 300 mg once every three weeks for the treatment of lung cancer, e.g., non-small cell lung cancer (NSCLC) (e.g., squamous NSCLC or non-squamous NSCLC). In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every three weeks to treat lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks to treat skin cancer, eg, Merkel cell cancer or melanoma. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of approximately 400 mg once every four weeks for the treatment of Merkel cell cancer. In other embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks for the treatment of melanoma. In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks to treat breast cancer, eg, triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of approximately 400 mg once every four weeks for the treatment of thyroid cancer, eg, anaplastic thyroid carcinoma (ATC). In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of approximately 400 mg once every four weeks to treat a neuroendocrine tumor (NET), e.g., atypical pulmonary carcinoid tumor or NET in the pancreas, gastrointestinal (GI) tract, or lung. . In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of approximately 400 mg once every four weeks to treat kidney cancer, e.g., renal cell carcinoma (e.g., clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma ( nccRCC)). In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks for the treatment of ovarian cancer. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks for the treatment of fallopian tube cancer. In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of approximately 400 mg once every four weeks for the treatment of colon and rectal cancer (e.g., highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable cancer). colon and rectum (MSS CRC)). In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of approximately 400 mg once every four weeks for the treatment of leukemia (eg, AML, eg, relapsed or refractory AML, or de novo AML). In some embodiments, an anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks for the treatment of myelodysplastic syndrome (MDS) (eg, high-risk MDS).

In some embodiments, the anti-PD-1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose (e.g., fixed dose) of about 100 mg to 600 mg, such as about 200 mg to 500 mg, such as about 100 mg to 300 mg, about 250 mg to 450 mg, about 300 mg to 400 mg, about 250 mg to 350 mg, about 350 mg to 450 mg, or about 100 mg, about 200 mg, about 300 mg, or approximately 400 mg. The administration schedule (eg, fixed dose schedule) may vary, for example, from once a week to once every 2, 3, 4, 5 or 6 weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg to 400 mg once every three weeks or once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every three weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every three weeks.

In another aspect of the invention, a method is provided for reducing the activity (eg, growth, survival, or viability, or whatever) of a hyperproliferative (eg, cancerous) cell. The method includes contacting a cell with an anti-PD-1 antibody molecule, such as an anti-PD-1 antibody molecule described herein. The method may be administered to a subject, eg, as part of a therapeutic protocol, eg, at a dose of about 300 mg to 400 mg of an anti-PD-1 antibody molecule once every three weeks or once every four weeks. In some embodiments, the dosage is approximately 300 mg of an anti-PD-1 antibody molecule once every three weeks. In other embodiments, the dosage is approximately 400 mg of an anti-PD-1 antibody molecule once every four weeks. The cancer cell can be, for example, a cancer cell described in this application, such as lung cancer (squamous), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous), stomach cancer, thyroid cancer, skin cancer , melanoma, nasopharyngeal cancer (eg, differentiated or undifferentiated metastatic or locally recurrent nasopharyngeal carcinoma), kidney cancer, neuroendocrine tumor (NET), ovarian cancer, fallopian tube cancer, colon and rectal cancer, or breast cancer. In some embodiments, the cancer is skin cancer, such as Merkel cell cancer or melanoma. In one embodiment, the cancer is Merkel cell cancer. In other embodiments, the cancer is melanoma. In other embodiments, the cancer is breast cancer, such as triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In other embodiments, the cancer is kidney cancer, eg, renal cell carcinoma (eg, clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC)). In other embodiments, the cancer is thyroid cancer, such as anaplastic thyroid carcinoma (ATC). In other embodiments, the cancer is a neuroendocrine tumor (NET), such as an atypical pulmonary carcinoid tumor or a NET in the pancreas, gastrointestinal (GI) tract, or lung. In some embodiments, the cancer is lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is colon and rectal cancer (CRC) (eg, highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC)). In some embodiments, the cancer is a leukemia (eg, acute myeloid leukemia (AML), eg, relapsed or refractory AML or de novo AML). In some embodiments, the cancer is a myelodysplastic syndrome (MDS) (eg, high risk MDS).

In another aspect of the invention, a composition (eg, one or more compositions or dosage forms) is provided that comprises an anti-PD-1 antibody molecule (eg, an anti-PD-1 antibody molecule as described herein). The present application also describes dosage forms, eg dosage forms and kits, eg therapeutic kits, which include an anti-PD-1 antibody molecule (eg, an anti-PD-1 antibody molecule as described herein). In some embodiments, the composition or dosage form comprises 300 mg or 400 mg of an anti-PD-1 antibody molecule (eg, an anti-PD-1 antibody molecule as described herein). In some embodiments, the composition or dosage form is administered or used once every three weeks or once every four weeks.

Also disclosed herein are methods and compositions comprising a combination of two, three or more therapeutic agents selected from one, two or all of the following categories (i)-(iii): (i) an agent that increases antigen presentation (e.g., tumor presentation). antigen); (ii) an agent that enhances an effector cell response (eg, activation and/or mobilization of B cells and/or T cells); or (iii) an agent that reduces tumor immunosuppression. In some embodiments, the combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein). In some embodiments, the anti-PD-1 antibody molecule is administered or administered at a constant or fixed dose.

Without being limited by theory, it is believed that therapeutic methods that enhance antitumor immunity work more effectively when the immune response is optimized by targeting multiple components at one or more stages of the immune response, for example, antitumor immune response. For example, methods that increase antigen presentation, such as activation and/or maturation of dendritic cells, are combined with methods that enhance cellular and humoral immune responses (eg, by stimulating, for example, activation, phagocytes and/or tumor-infiltrating lymphocytes (eg, NK cells and T cells)), while blocking tumor immunosuppressive signals (eg, by increasing macrophage polarization, increasing T _reg depletion, and/or decreasing myeloid suppressor cells (MSCs)) may lead to more effective and/or prolonged therapeutic response. reactions. Thus, the present application discloses combination therapies that optimize one, two, or all of: (i) antigen presentation, e.g., increased antigen presentation (e.g., by increasing one or more of dendritic cell activity or maturation, antigen uptake, or antigen processing ); (ii) effector cell response, eg, increased effector cell response (eg, increased activation and/or mobilization of B cells and/or T cells, eg, in a lymph node); or (iii) tumor immunosuppression, eg decreasing tumor immunosuppression (eg increasing T cell infiltration and tumor cell killing). The combinations described herein can provide superior benefits, for example in the treatment of a disorder, such as enhanced antitumor activity, reduced toxicity, and/or reduced side effects, as compared to the use of therapeutic agents alone in combination. For example, one or more therapeutic agents in combination may be administered at a lower dose or for a shorter period of use than would be necessary to achieve the same therapeutic effect compared to monotherapy. Thus, compositions and methods for treating cancer and other immunological disorders using the above combination therapy are disclosed.

Thus, in one aspect of the invention, there is provided a method of treating (eg, inhibiting, reducing, mitigating, or preventing) a disorder, eg, a hyperproliferative condition or disorder (eg, cancer) in a subject. The method includes administering to a subject a combination of two, three, or more therapeutic agents selected from one, two, or all of the following categories (i)-(iii): (i) an agent that increases antigen (eg, tumor antigen) presentation; (ii) an agent that enhances an effector cell response (eg, activation and/or mobilization of B cells and/or T cells); or (iii) an agent that reduces tumor immunosuppression such that a disorder, eg, a hyperproliferative condition, or a disorder (eg, cancer) is treated. In some embodiments, the combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein). The cancer being treated may be, for example, the cancer described herein, such as lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer, skin cancer , melanoma, nasopharyngeal cancer (eg, differentiated or undifferentiated metastatic or locally recurrent nasopharyngeal carcinoma), kidney cancer, neuroendocrine tumor (NET), or breast cancer. In some embodiments, the cancer is skin cancer, such as Merkel cell cancer or melanoma. In one embodiment, the cancer is Merkel cell cancer. In other embodiments, the cancer is melanoma. In other embodiments, the cancer is breast cancer, such as triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In other embodiments, the cancer is kidney cancer, eg, renal cell carcinoma (eg, clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC)). In other embodiments, the cancer is thyroid cancer, such as anaplastic thyroid carcinoma (ATC). In other embodiments, the cancer is a neuroendocrine tumor (NET), such as an atypical pulmonary carcinoid tumor or a NET in the pancreas, gastrointestinal (GI) tract, or lung. In some embodiments, the cancer is lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, the cancer is ovarian cancer. In some embodiments, the implementation of the cancer is cancer of the fallopian tube. In some embodiments, the cancer is colon and rectal cancer (CRC) (eg, highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC)). In some embodiments, the cancer is a leukemia (eg, acute myeloid leukemia (AML), eg, relapsed or refractory AML, or de novo AML). In some embodiments, the cancer is a myelodysplastic syndrome (MDS) (eg, high risk MDS).

In another aspect of the invention, there is provided a method for reducing the activity (eg, growth, survival, or viability, or whatever) of a hyperproliferative (eg, cancerous) cell. The method includes contacting a cell with a combination of two, three, or more therapeutic agents selected from one, two, or all of the following categories (i)-(iii): (i) an agent that increases antigen (eg, tumor antigen) presentation; (ii) an agent that enhances an effector cell response (eg, activation and/or mobilization of B cells and/or T cells); or (iii) an agent that reduces tumor immunosuppression, resulting in a decrease in the activity of a hyperproliferative cell. In some embodiments, the combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein). The method can be applied to a subject, for example, as part of a therapeutic protocol. The cancer cell can be, for example, a cancer cell described in this application, such as lung cancer (squamous), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous), stomach cancer, thyroid cancer, skin cancer , melanoma, nasopharyngeal cancer (eg, differentiated or undifferentiated metastatic or locally recurrent nasopharyngeal carcinoma), kidney cancer, neuroendocrine tumor (NET), ovarian cancer, fallopian tube cancer, colon and rectal cancer, or breast cancer. In some embodiments, the cancer is skin cancer, such as Merkel cell cancer or melanoma. In one embodiment, the cancer is Merkel cell cancer. In other embodiments, the cancer is melanoma. In other embodiments, the cancer is breast cancer, such as triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In other embodiments, the cancer is kidney cancer, eg, renal cell carcinoma (eg, clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC)). In other embodiments, the cancer is thyroid cancer, such as anaplastic thyroid carcinoma (ATC). In other embodiments, the cancer is a neuroendocrine tumor (NET), such as an atypical pulmonary carcinoid tumor or a NET in the pancreas, gastrointestinal (GI) tract, or lung. In some embodiments, the cancer is lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is colon and rectal cancer (CRC) (eg, highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC)). In some embodiments, the cancer is a leukemia (eg, acute myeloid leukemia (AML), eg, relapsed or refractory AML or de novo AML). In some embodiments, the cancer is a myelodysplastic syndrome (MDS) (eg, high risk MDS).

In some embodiments of the methods disclosed herein, the method further comprises determining a subject's level of immunocellular (eg, T-cell) infiltrate (eg, tumor-infiltrating lymphocyte (TIL) level). In one embodiment, the level of immunocellular infiltrate is determined in vivo , for example, by a non-invasive method (for example, by detecting an antibody to a T-cell marker labeled with a detectable label using a suitable imaging method, for example, a positron emission tomography (PET) scan). In other embodiments, the level of immunocellular infiltrate is determined in a sample (eg, tumor biopsy) obtained from a subject (eg, using immunohistochemical methods). In embodiments sensitive to a low or undetectable level of tumor infiltrate in a subject, one or more agents of categories (i) and/or (ii) are administered. In other embodiments sensitive to a detectable level or elevated level of tumor infiltrate in a subject, one or more agents of category (iii) are administered. The detection steps can also be used, for example, to monitor the effectiveness of a therapeutic agent described in this application. For example, the detection step may be used to monitor the efficacy of category (i), (ii) and/or (iii) therapeutics.

In another aspect of the invention, a composition (e.g., one or more compositions or dosage forms) is provided that comprises a combination of two, three, or more therapeutic agents selected from one, two, or all of the following categories (i)-(iii): (i) an agent that increases the presentation of an antigen (eg, tumor antigen); (ii) an agent that enhances an effector cell response (eg, activation and/or mobilization of a B cell and/or T cell); or (iii) an agent that reduces tumor immunosuppression. In some embodiments, the combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein).

In yet another aspect of the invention, a composition (eg, one or more compositions or dosage forms as described herein) is provided for use in the treatment of a disorder, eg, cancer. In embodiments, the composition for use includes a combination of two, three, or more therapeutic agents selected from one, two, or all of the following categories (i)-(iii): (i) an agent that increases antigen (eg, tumor antigen) presentation; (ii) an agent that enhances an effector cell response (eg, activation and/or mobilization of a B cell and/or T cell); or (iii) an agent that reduces tumor immunosuppression. In some embodiments, the combination used comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein). The cancer may be, for example, the cancer described in this application, such as lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer, skin cancer, melanoma , nasopharyngeal cancer (eg, differentiated or undifferentiated metastatic or locally recurrent nasopharyngeal carcinoma), kidney cancer, neuroendocrine tumor (NET), ovarian cancer, fallopian tube cancer, colon and rectal cancer, or breast cancer. In some embodiments, the cancer is skin cancer, such as Merkel cell cancer or melanoma. In one embodiment, the cancer is Merkel cell cancer. In other embodiments, the cancer is melanoma. In other embodiments, the cancer is breast cancer, such as triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In other embodiments, the cancer is kidney cancer, eg, renal cell carcinoma (eg, clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC)). In other embodiments, the cancer is thyroid cancer, such as anaplastic thyroid carcinoma (ATC). In other embodiments, the cancer is a neuroendocrine tumor (NET), such as an atypical pulmonary carcinoid tumor or a NET in the pancreas, gastrointestinal (GI) tract, or lung. In some embodiments, the cancer is lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is colon and rectal cancer (CRC) (eg, highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC)). In some embodiments, the cancer is a leukemia (eg, acute myeloid leukemia (AML), eg, relapsed or refractory AML, or de novo AML). In some embodiments, the cancer is a myelodysplastic syndrome (MDS) (eg, high risk MDS).

Dosage forms, such as dosage forms, and kits, such as therapeutic kits, which include a combination of two, three or more therapeutic agents selected from one, two or all of the following categories (i)-(iii): (i) an agent that increases presentation of an antigen (eg, tumor antigen); (ii) an agent that enhances an effector cell response (eg, activation and/or mobilization of a B cell and/or T cell); or (iii) an agent that reduces tumor immunosuppression, resulting in a decrease in activity in the cell, and (optionally) the instructions for use are also disclosed. In some embodiments, the combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein).

Combinations of therapeutic agents disclosed in this application include two or more therapeutic agents described in this application. Therapeutic agents in combination may be in the same category, e.g., two or more category (i) therapeutic agents, or may include at least one agent from two or more categories (e.g., a category (i) therapeutic agent in combination with therapeutic agent of category (ii)), as described below. Some therapeutic agents may fall into two or more categories from categories (i)-(iii). For example, a therapeutic agent (eg, GITR agonist, IDO antagonist, TGF-b inhibitor, among others) can act as a therapeutic agent in many categories.

Additional features or embodiments of the methods, compositions, dosage forms, and kits described herein include one or more of the following:

Combinations

In some embodiments, the combination includes one, two, three, four or more therapeutic agents that increase the presentation of an antigen (eg, tumor antigen) (referred to herein as an "antigen-presenting combination"). In some embodiments, the antigen-presenting combination comprises one or more of: an agent that increases antigen presentation (eg, a vaccine, eg, a cell- or antigen-based vaccine); an agent that increases the lysis of tumor cells (eg, oncolytic virus); an agent that stimulates (e.g., activates) a phagocyte, e.g., a type I interferon (IFN) activator (e.g., a TLR agonist, a RIG-I-like receptor (RLR) agonist), and/or an agent that activates and/or recruits a dendritic cell or macrophage (eg, macrophage I), for example, bi- or trispecific cell activators.

In some embodiments, the antigen presenting combination comprises one, two, three, four, five or more therapeutic agents selected from: (i) an interferon gene stimulator agonist (STING agonist), (ii) a Toll-like receptor (TLR) agonist (e.g., a TLR-3, -4, -5, -7, -8, or -9 agonist), (iii) a TIM-3 modulator (e.g., an anti-TIM-3 antibody molecule), (iv) a vascular endothelial growth factor receptor (VEGFR) inhibitor ), (v) c-Met inhibitor, (vi) TGFb inhibitor (e.g., anti-TGFb antibodies), (vii) IDO/TDO inhibitor, (viii) A2AR antagonist, (ix) oncolytic virus, (x) vaccines (e.g., scaffold vaccines) or (xi) a bi- or tri-specific cell activator. Any combination of the above means (i)-(xi) can be used in an antigen-presenting combination. In one embodiment, the antigen presenting combination comprises a STING agonist. In another embodiment, the antigen-presenting combination comprises a TLR agonist (eg, a TLR7 agonist). In another embodiment, the antigen presenting combination comprises a STING agonist and a TLR agonist (eg, a TLR7 agonist). In some embodiments, the antigen presenting combination is selected from a STING agonist, a TLR agonist, an A2AR antagonist, or an oncolytic virus, or a combination thereof, and optionally one or more of (iii)-(vii) or (x)-(xi). In some embodiments, the antigen-presenting combination is selected from a STING agonist or a TLR agonist, or a combination thereof, and optionally one or more of (iii)-(xi). In another embodiment, the antigen-presenting combination comprises a STING agonist, a TLR agonist (eg, a TLR7 agonist), and a TIM-3 modulator (eg, a TIM-3 inhibitor). In another embodiment, the antigen presenting combination comprises a STING agonist, a TLR agonist (eg, a TLR7 agonist), and a VEGFR inhibitor. In another embodiment, the antigen presenting combination comprises a STING agonist, a TLR agonist (eg, a TLR7 agonist), and a c-MET inhibitor. In other embodiments, the antigen-presenting combination comprises an oncolytic virus. In other embodiments, the antigen-presenting combination comprises an oncolytic virus and a cytokine, eg, an oncolytic virus expressing one or more of GM-CSF or M-CSF (eg, CSF1 or CSF2). In some embodiments, the antigen-presenting combination comprises a bi- or trispecific cell activator, for example, a bi- or trispecific anti-CD47 and CD19 antibody molecule, with or without an Fc domain. In some embodiments, the antigen-presenting combination comprises a TGFb inhibitor (eg, an anti-TGFb antibody). In other embodiments, the antigen presenting combination comprises an IDO/TDO inhibitor. In other embodiments, the antigen-presenting combination comprises an A2AR antagonist. In other embodiments, the antigen presenting combination comprises a vaccine (eg IL-2 in combination with MUC1 or a dendritic cell based vaccine (eg Provenge®)). In other embodiments, the antigen-presenting combination comprises a vaccine and a TLR agonist (eg, a TLR agonist as described herein). In a certain embodiment, the antigen presenting combination comprises a vaccine and a STING agonist. In a particular embodiment, the antigen-presenting combination comprises a vaccine, a STING agonist, and a TLR agonist.

In some embodiments, the combination includes one, two, three, four, five, or more therapeutic agents that enhance the response of an effector cell (referred to herein as an "effector cell combination"). In some embodiments, the effector cell combination comprises a lymphocyte activator, such as an NK cell activator and/or a T cell activator. In some embodiments, the effector cell combination activates (eg, deinhibits) a tumor infiltrating lymphocyte (TIL), such as an NK cell or a T cell. In some embodiments, the effector cell combination comprises an NK cell modulator selected from a modulator (eg, an antibody molecule) of an NK receptor (eg, a modulator of one or more of NKG2A, KIR3DL, NKp46, MICA, or CEACAM1); an interleukin or interleukin variant (eg, cytokine IL-2, IL-15, IL-21, IL-13R, or IL-12, or a variant or combination thereof); a bi- or trispecific cell activator (eg, NKG2A and CD138 bispecific antibody molecules or CD3 and TCR bispecific antibody molecules); NK cell therapy; or a vaccine that includes NK cells and an antigen/immune stimulant. In some embodiments, the effector cell combination comprises an immunomodulator (eg, one or more of a costimulatory molecule activator or an immune checkpoint molecule inhibitor, as described herein). In some embodiments, the effector cell combination comprises a T cell modulator selected from a checkpoint inhibitor inhibitor (e.g., an inhibitor of one or more of: PD-1, PD-L1, TIM-3, LAG-3, VISTA, DKG-α, B7-H3, B7-H-4, TIGIT, CTLA-4, BTLA, CD160, TIM1, IDO, LAIR1, IL-12, or combinations thereof, such as a PD-1 and TIM-3 inhibitor, or a PD-1 inhibitor and LAG-3). In one embodiment, the checkpoint inhibitor inhibitor is an antibody molecule (eg, a mono- or bispecific antibody or fragment thereof, as described herein). For example, a checkpoint inhibitor is an anti-PD-1, PD-L1, TIM-3, LAG-3, VISTA, B7-H-4, CTLA-4, or TIGIT antibody molecule, or any combination thereof (e.g., a combination as described in this application). In some embodiments, the effector cell combination comprises a T cell modulator selected from an agonist or an activator of a costimulatory molecule. In one embodiment, the costimulatory molecule agonist is selected from an agonist (e.g., agonist antibody or antigen-binding fragment thereof, or soluble fusion construct) GITR, OX40, ICOS, SLAM (e.g., SLAMF7), HVEM, LIGHT, CD2, CD27, CD28, CD, ICAM -1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137), CD30, CD40, BAFFR, CD7, NKG2C, NKp80, CD160, B7-H3 or CD83 ligand. In other embodiments, the effector cell combination comprises a bispecific T cell activator (eg, a bispecific antibody molecule that binds to CD3 and a tumor antigen (eg, EGFR, PSCA, PSMA, EpCAM, HER2, among others).

In some embodiments, the effector cell combination comprises one, two, three, four, five, or more therapeutic agents selected from: (i) a GITR modulator (e.g., a GITR agonist), (ii) a PD-1 inhibitor (e.g., an anti- PD-1 as described herein), (iii) PD-L1 inhibitor, (iv) IAP (apoptosis inhibitory protein), (v) EGFR (epidermal growth factor receptor) inhibitor, (vi) rapamycin target inhibitor ( mTOR), (vii) IL-15 or a variant thereof, (viii) a CTLA-4 inhibitor, (ix) a bispecific T cell activator (e.g., a bispecific antibody molecule that binds to CD3 and a tumor antigen (e.g., EGFR, PSCA, PSMA, EpCAM, HER2, among others), (x) a CD40 agonist (e.g., anti-CD40 antibody molecules), (xi) an OX40 agonist (e.g., anti-OX40 antibody molecules), or (xii) a CD27 agonist (e.g., anti-CD27 antibody molecules ) Any combination of the above agents can be used in an effector cell combination. In one embodiment, the effector cell combination comprises a GITR agonist. In another embodiment, the effector cell combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein). In another embodiment, the effector cell combination comprises a PD-L1 inhibitor. In other embodiments, the effector cell combination comprises a GITR agonist and a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein). In other embodiments, the effector cell combination comprises a GITR agonist and a PD-L1 inhibitor. In other embodiments, the effector cell combination comprises a GITR agonist, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein) and a PD-L1 inhibitor. In other embodiments, the effector cell combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein) and a PD-L1 inhibitor. In one embodiment, the effector cell combination comprises a GITR agonist and an IAP inhibitor. In another embodiment, the effector cell combination comprises a GITR agonist and an EGFR inhibitor inhibitor. In yet another embodiment, the effector cell combination comprises a GITR agonist and an mTOR inhibitor inhibitor. In one embodiment, the effector cell combination comprises IL-15 or a variant thereof. In one embodiment, the effector cell combination comprises a CTLA-4 inhibitor. In one embodiment, the effector cell combination comprises a bispecific T cell activator (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 embodiment, the effector combination the cell comprises a CD40 agonist (e.g., an anti-CD40 antibody molecule) In one embodiment, the effector cell combination comprises an OX40 agonist (e.g., an anti-OX40 antibody molecule) In one embodiment, the effector cell combination comprises a CD27 agonist (e.g., an anti-CD27 antibody molecule) .

In some embodiments, the combination includes one, two, three, four, five, or more therapeutic agents that reduce tumor immunosuppression (referred to herein as "anti-tumor immunosuppression combination"). In some embodiments, the combination modulates the activity or level of one or more of T _reg , macrophage 2, or MSCs. In some embodiments, the combination increases one or more of M2 polarization, T _reg depletion, or T cell recruitment. In some embodiments, the antitumor immunosuppression combination comprises one, two, three, four, five, or more therapeutic agents selected from: (i) an immunomodulator (e.g., one or more of: immune checkpoint molecules (e.g. one or more of PD-1, PD-L1, LAG-3, TIM-3 or CTLA-4) as described herein), (ii) a CSF-1/1R inhibitor (e.g. , a macrophage colony stimulating factor (M-CSF) inhibitor), (iii) an IL-17 inhibitor, (iv) an IL-1β inhibitor, (v) a CXCR2 inhibitor, (vi) a phosphoinositide 3-kinase inhibitor (PI3K, e.g. PI3Kγ or PI3Kδ), (vii) BAFF-R inhibitor, (viii) MALT-1/BTK inhibitor, (ix) JAK inhibitor, (x) CRTH2 inhibitor, (xi) VEGFR inhibitor, (xiii) IL-15 or variant thereof, ( xiv) a CTLA-4 inhibitor, (xv) an IDO/TDO inhibitor, (xvi) an A2AR antagonist, (xvii) a TGFb inhibitor, or (xviii) a PFKFB3 inhibitor. In some embodiments, the immunomodulator is an inhibitor of an immune checkpoint molecule (e.g., an inhibitor of PD-1, PD-L1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, -3 and/or -5) or CTLA- 4 or any combination of them). Any combination of the above agents can be used in combination tumor immunosuppression. In one embodiment, the anti-tumor immunosuppression combination comprises one, two, three, four, five or more therapeutic agents selected from a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule as described herein), a PD-L1 inhibitor , LAG-3 inhibitor, TIM-3 modulator (eg, TIM-3 inhibitor), GITR agonist, CSF-1/1R inhibitor (eg, M-CSF inhibitor), IL-17 inhibitor, IL-1β inhibitor, or CXCR2 inhibitor. In one embodiment, the anti-tumor immunosuppression combination comprises one, two, or all of a CSF-1/1R inhibitor (eg, an M-CSF inhibitor), an IL-17 inhibitor, an IL-1β inhibitor. In one embodiment, the anti-tumor immunosuppression combination comprises an IL-17 inhibitor, a CXCR2 inhibitor, a CRTH2 inhibitor, an A2AR antagonist, or a PFKFB3 inhibitor, or a combination thereof.

In some embodiments, the combination includes one or more antigen-presenting combination therapeutic agents. In other embodiments, the combination comprises one or more effector cell combination therapeutic agents. In other embodiments, the combination comprises one or more combination therapeutic agents for tumor immunosuppression. In other embodiments, the combination comprises one or more antigen-presenting combination therapeutics and one or more effector cell combination therapeutics. In other embodiments, the combination comprises one or more antigen-presenting combination therapeutics and one or more tumor immunosuppression combination therapeutics. In other embodiments, the combination comprises one or more antigen presenting combination therapeutics, one or more effector cell combination therapeutics, and one or more tumor immunosuppression combination therapeutics. In other embodiments, the combination comprises one or more antigen presenting combination therapeutics, one or more effector cell combination therapeutics, and one or more tumor immunosuppression combination therapeutics.

In some embodiments, the combination includes:

(i) one or more antigen-presenting combination therapeutic agents selected from one, two or all of a STING agonist, a TLR agonist (eg, a TLR7 agonist), or a TIM-3 modulator (eg, a TIM-3 inhibitor);

(ii) one or more effector cell combination therapeutics selected from one, two, or all of a GITR modulator (eg, a GITR agonist), a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein), or a PD-L1 inhibitor;

(iii) one or more antitumor immunosuppression combination therapeutic agents 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-1β inhibitor:

(iv) a combination of (i) and (ii);

(v) a combination of (i) and (iii);

(vi) a combination of (ii) and (iii); or

(vii) a combination of (i), (ii) and (iii).

The combination can be used to treat cancer as described herein, such as lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer, skin cancer, melanoma (eg, advanced melanoma), nasopharyngeal cancer, kidney cancer, neuroendocrine tumor (NET), ovarian cancer, fallopian tube cancer, colon and rectal cancer, or breast cancer. In some embodiments, the cancer is skin cancer, such as Merkel cell cancer or melanoma. In one embodiment, the cancer is Merkel cell cancer. In other embodiments, the cancer is melanoma. In other embodiments, the cancer is breast cancer, such as triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In other embodiments, the cancer is kidney cancer, eg, renal cell carcinoma (eg, clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC)). In other embodiments, the cancer is thyroid cancer, such as anaplastic thyroid carcinoma (ATC). In other embodiments, the cancer is a neuroendocrine tumor (NET), such as an atypical pulmonary carcinoid tumor or a NET in the pancreas, gastrointestinal (GI) tract, or lung. In some embodiments, the cancer is lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is colon and rectal cancer (CRC) (eg, highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC)). In some embodiments, the cancer is a leukemia (eg, acute myeloid leukemia (AML), eg, relapsed or refractory AML or de novo AML). In some embodiments, the cancer is a myelodysplastic syndrome (MDS) (eg, high risk MDS).

In other embodiments, the combination comprises a therapeutic agent from an antigen-presenting combination (e.g., one or more of a STING agonist, a TLR agonist, a vaccine, or an oncolytic virus) in combination with a therapeutic agent from an effector cell combination and/or an anti-tumor immunosuppression combination (e.g., an inhibitor of an anti-tumor immunosuppression checkpoint, e.g., a PD-1, PD-L1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, -3 and/or -5) inhibitor, or CTLA-4 inhibitor, or any combination thereof. one or more of a STING agonist, a TLR agonist, a vaccine, or an oncolytic virus is used in combination with an anti-PD-1 antibody molecule as described herein In one embodiment, the STING agonist and/or vaccine is used in combination with an anti-PD antibody molecule -1 as described herein In one embodiment, an oncolytic virus is used in combination with an anti-PD-1 antibody molecule as described herein. turnout. The combination can be used to treat cancer as described herein, such as lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer, skin cancer, melanoma (eg, advanced melanoma), nasopharyngeal cancer, kidney cancer, neuroendocrine tumor (NET), ovarian cancer, fallopian tube cancer, colon and rectal cancer, or breast cancer. In some embodiments, the cancer is skin cancer, such as Merkel cell cancer or melanoma. In one embodiment, the cancer is Merkel cell cancer. In other embodiments, the cancer is melanoma. In other embodiments, the cancer is breast cancer, such as triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In other embodiments, the cancer is kidney cancer, eg, renal cell carcinoma (eg, clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC)). In other embodiments, the cancer is thyroid cancer, such as anaplastic thyroid carcinoma (ATC). In other embodiments, the cancer is a neuroendocrine tumor (NET), such as an atypical pulmonary carcinoid tumor or a NET in the pancreas, gastrointestinal (GI) tract, or lung. In some embodiments, the cancer is lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is fallopian tube cancer. In some embodiments, the cancer is colon and rectal cancer (CRC) (eg, highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC)). In some embodiments, the cancer is a leukemia (eg, acute myeloid leukemia (AML), eg, relapsed or refractory AML or de novo AML). In some embodiments, the cancer is a myelodysplastic syndrome (MDS) (eg, high risk MDS).

In some embodiments, the combination includes a combination of therapeutic agents as set forth in the section entitled "Examples of Antigen Presenting Combinations, Effector Cell Combinations, and Anti-Tumor Immunosuppression Combinations" provided in the Detailed Description.

In some embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and an IAP inhibitor, (S)-N-(S)-1-cyclohexyl -2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Patent 8,552,003. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Pat. No. 8,552,003 is administered at a dose of 200 mg to 400 mg (e.g. 300 mg), e.g. once a week, for example, orally. In some embodiments, the combination is used to treat the cancer described herein, e.g., colon and rectal cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC)) or breast cancer (e.g., triple negative breast cancer (e.g., TNRMZH)).

In other embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT Publication WO 2014/085318. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 2 mg to 8 mg (e.g., a 5 mg dose), e.g., once a week eg orally. In some embodiments, the combination is used to treat the cancer described herein, e.g., colon and rectal cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC)) or breast cancer (e.g., triple negative breast cancer (e.g., TNRMZH)).

In other embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a DAC inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT Publication WO 2014/072493. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, a DAC inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO 2014/072493, is administered at a dose of 5 mg to 15 mg (e.g., 10 mg), e.g., three times a week ( for example, according to the scheme of administration week after week), for example, orally. In some embodiments, the combination is used to treat the cancer described herein, e.g., colon and rectal cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC)) or breast cancer (e.g., triple negative breast cancer (e.g., TNRMZh)).

In other embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and an IL-1β inhibitor, canakinumab, or a compound disclosed in PCT publication WO 2002 /16436. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, the IL-1β inhibitor, canakinumab, or a compound disclosed in PCT publication WO 2002/16436, is administered at a dose of 50 mg to 150 mg (e.g., 100 mg dose), e.g., once every eight weeks, e.g. , subcutaneously. In some embodiments, the combination is used to treat the cancer described herein, e.g., colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS CRC)), lung cancer (e.g., non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple-negative breast cancer (eg, TNBC)).

In other embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and an IL-17 inhibitor, CJM112, or a compound disclosed in PCT publication WO 2014 /122613. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, an IL-17 inhibitor, CJM112, or a compound disclosed in PCT publication WO 2014/122613, is administered at a dose of 10 mg to 50 mg (e.g., 25 mg dose), e.g., once every four weeks, e.g. , intravenously. In some embodiments, the combination is used to treat the cancer described herein, for example, colon and rectal cancer (eg, MSS CRC), lung cancer (eg, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple negative breast cancer (e.g. TNBC)).

In other embodiments, the combination includes an immune checkpoint molecule inhibitor, such as a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein) and a MEK inhibitor or trametinib. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, the MEK inhibitor or trametinib is administered at a dose of 0.2 mg to 1 mg (eg, at a dose of 0.5 mg), eg, once a day, eg, orally. In some embodiments, the combination is used to treat the cancer described herein, e.g., colon and rectal cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC)) or breast cancer (e.g., triple negative breast cancer (e.g., TNRMZh)).

In some embodiments, the combination includes an immune checkpoint molecule inhibitor, such as a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein), a BRAF inhibitor or dabrafenib, and a MEK inhibitor or trametinib. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 8 weeks, for example, intravenously. In some embodiments, the BRAF inhibitor or dabrafenib is administered at a dose of 50 mg to 250 mg (eg, 150 mg) twice daily, eg, orally. In some embodiments, the MEK inhibitor or trametinib is administered at a dose of 1 mg to 3 mg (eg, at a dose of 2 mg), eg, once a day, eg, orally. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered, e.g., at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, following administration of a BRAF inhibitor or dabrafenib, a MEK inhibitor or trametinib, or both, for eg 2-8 weeks, eg 4 weeks. In some embodiments, the combination is used to treat the cancer described herein, eg, skin cancer (eg, melanoma, eg, unresectable or metastatic melanoma). In some embodiments, the combination is used to treat cancer that has a BRAF mutation, such as a BRAF V600 mutation. In other embodiments, the combination is used to treat cancer in a subject having an elevated serum lactate dehydrogenase (LDH) level relative to a reference LDH level. In some embodiments, the combination is used to treat a subject who has unresectable or metastatic melanoma having a BRAF mutation (eg, a BRAF V600 mutation) and has an elevated serum LDH level compared to a reference serum LDH level.

In other embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and an EGFR inhibitor, (R,E)-N-(7-chloro- 1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT Publications WO 2013/184757. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, is administered at a dose of 10 mg to 50 mg (for example, at a dose of 25 mg), for example, once every day (for example, on day 1 to day 10 of the first course of administration), for example, orally. In some embodiments, the combination is used to treat the cancer described herein, e.g., colon and rectal cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC)) or breast cancer (e.g., triple negative breast cancer (e.g., TNRMZH)).

In other embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor). ), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT Publication WO 2005/073224. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 50 mg to 1000 mg (e.g., 100 mg, 150 mg, 300 mg, 600 mg , 900 mg), eg, daily, according to a 7-day schedule every 7 days, or once a week, eg, orally. In some embodiments, the combination is used to treat the cancer described herein, e.g., brain cancer (e.g., glioblastoma multiforme (GBM)), breast cancer (e.g., triple-negative breast cancer (e.g., TNBC)) or pancreatic cancer. glands.

In other embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase inhibitor. 3 (PFKFB3), PFK-158, or a compound disclosed in PCT Publication WO 2013/148228. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiments, the combination is used to treat the cancer described herein, for example, ovarian cancer, fallopian tube cancer, or colorectal cancer (CRC).

In one embodiment, the combination comprises an inhibitor of an immune checkpoint molecule, eg, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein) and a chemotherapeutic agent, eg, paclitaxel (eg, paclitaxel-albumin). In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., a 400 mg dose), e.g., once every 4 weeks, for example, intravenously. In some embodiment, a chemotherapeutic agent, for example, paclitaxel (for example, paclitaxel-albumin), is administered at a dose of 50 mg/m ² to 200 mg/m ² (for example, at a dose of 100 mg/m ² ), for example, weekly on the Day 1, Day 8 and Day 15 every four weeks (qw3/4), for example, intravenously. In some embodiments, the combination is used to treat the cancer described herein, eg, breast cancer (eg, HER2-negative breast cancer).

In other embodiments, the combination comprises an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 100 mg to 600 mg (e.g., a 100 mg or 300 mg dose), e.g., once every 3 weeks, for example, intravenously. In some embodiments, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of 0.1 mg/kg to 15 mg/kg, such as 0.1 mg/kg to 6 mg/kg, or 0.3 mg/kg to 3 mg/kg (for example, at a dose of 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 6 mg/kg kg, 10 mg/kg or 15 mg/kg), for example once every three weeks, for example intravenously. In some embodiments, the combination is used to treat the cancer described herein, e.g., pancreatic cancer, colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS-CRC)), lung cancer (e.g., non-small cell cancer lung), breast cancer (eg, triple negative breast cancer), liver cancer (eg, hepatocellular carcinoma), prostate cancer, or kidney cancer (eg, clear cell renal cell carcinoma).

In other embodiments, the combination comprises an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a PD-L1 inhibitor or an anti-PD-L1 antibody molecule disclosed in the Application Publication. for US Patent 2016/0108123. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 100 mg to 600 mg (e.g., 100 mg or 300 mg), e.g., once every 3 weeks, for example, intravenously. In some embodiments, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US Patent Application Publication 2016/0108123 is administered at a dose of 10 mg to 2000 mg (e.g., 20 mg to 1600 mg or 80 mg to 1200 mg (eg, at a dose of 20 mg, 80 mg, 240 mg, 800 mg, or 1200 mg), eg, once every three weeks or once every six weeks, eg, intravenously In some embodiments, the combination is used to treat cancer described herein, e.g., a solid tumor, e.g., lung cancer (e.g., non-small cell lung cancer), breast cancer (e.g., triple negative breast cancer), uterine cancer (e.g., endometrial cancer), or thyroid cancer (e.g., , anaplastic thyroid carcinoma).

In other embodiments, the combination includes an immune checkpoint molecule inhibitor, eg, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein) and a chemotherapeutic agent (eg, a hypomethylating agent, eg, decitabine). In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., 300 mg to 500 mg, e.g., 400 mg), for example, once every 4 weeks, for example, intravenously. In some embodiments, the chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered at a dose of 5 mg/m ² to 50 mg/m ² (e.g., 10 mg/m ² to 30 mg/m ² , e.g., dose of 20 mg/m ² ), for example, daily, for example, for 1, 2, 3, 4, 5, 6, 7, 8 or more days, for example, intravenously. In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered on day 8 of a 28 day course and a chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered on days 1, 2, 3, 4 and 5 of the 28-day course. In some embodiments, the combination is used to treat the cancer described herein, e.g., hemoblastosis, e.g., leukemia (e.g., acute myeloid leukemia (AML), e.g., relapsed or refractory AML or de novo AML), or myelodysplastic syndrome (MDS) (eg high-risk MDS).

In other embodiments, the combination comprises an inhibitor of an immune checkpoint molecule, e.g., a TIM-3 inhibitor or an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274, and a chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) . In some embodiments, an inhibitor of an immune checkpoint molecule, such as a TIM-3 inhibitor or an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274, is administered at a dose of 50 mg to 500 mg (e.g., 100 mg up to 400 mg, for example, at a dose of 240 mg), for example, once every 2 weeks, for example, intravenously. In some embodiments, the chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered at a dose of 5 mg/m ² to 50 mg/m ² (e.g., 10 mg/m ² to 30 mg/m ² , e.g., dose of 20 mg/m ² ), for example, daily, for example, for 1, 2, 3, 4, 5, 6, 7, 8 or more days, for example, intravenously. In some embodiments, an inhibitor of an immune checkpoint molecule, such as a TIM-3 inhibitor or an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274, is administered on days 8 and 22 of a 28 day course and a chemotherapeutic agent (e.g. , a hypomethylating agent such as decitabine) is administered on days 1, 2, 3, 4 and 5 of a 28-day course. In some embodiments, the combination is used to treat the cancer described herein, e.g., hemoblastosis, e.g., leukemia (e.g., acute myeloid leukemia (AML), e.g., relapsed or refractory AML or de novo AML) or myelodysplastic syndrome (MDS) ( e.g. high-risk MDS).

In other embodiments, the combination comprises a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein), a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274 ) and a chemotherapeutic agent (eg, a hypomethylating agent, eg, decitabine). In some embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 600 mg (e.g., 300 mg to 500 mg, e.g., 400 mg), e.g., once at 4 weeks, for example, intravenously. In some embodiments, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) is administered at a dose of 20 mg to 400 mg (e.g., 40 mg to 200 mg or 50 mg to 100 mg, eg at a dose of 80 mg), eg once every 2 weeks, eg intravenously. In some embodiments, the chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered at a dose of 5 mg/m ² to 50 mg/m ² (e.g., 10 mg/m ² to 30 mg/m ² , e.g., dose of 20 mg/m ² ), for example, daily, for example, for 1, 2, 3, 4, 5, 6, 7, 8 or more days, for example, intravenously. In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered on day 8 of a 28-day course, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in U.S. Patent Application Publication 2015/ 0218274) is administered on days 8 and 22 of the 28 day course and the chemotherapeutic agent (eg decitabine) is administered on days 1, 2, 3, 4 and 5 of the 28 day course. In some embodiments, the combination is used to treat the cancer described herein, e.g., hemoblastosis, e.g., leukemia (e.g., acute myeloid leukemia (AML), e.g., relapsed or refractory AML or de novo AML), or myelodysplastic syndrome (MDS) (eg high-risk MDS).

The combinations disclosed in this application may be administered together in one composition or administered separately in two or more different compositions, for example, compositions or dosage forms, as described in this application. The administration of the therapeutic agents may be in any order. The first agent and additional agents (eg, second, third agents) may be administered by the same route of administration or by different routes of administration. For example, the first therapeutic agent may be administered simultaneously, before or after the additional agent. In some embodiments, the first agent is administered topically, e.g., a therapeutic agent of any of categories (i)-(iii) may be associated with a tumor-targeting agent, e.g., a tumor-targeting antibody (e.g., to form an antibody-drug conjugate). agent), or any other carrier (eg, a dosage form, such as a targeted dosage form) such that the administration of the first agent is localized to the desired area, for example, a tumor site (eg, a region rich in dendritic cells). In one embodiment, the therapeutic agent is an antigen (eg, a vaccine, eg, an in situ cancer vaccine) that interacts in a targeted manner with the tumor environment, resulting in dendritic cell activation. The therapeutic agent can also be administered topically, eg, administered to the site of the tumor (eg, intratumoral or peritumoral administration). Localized delivery or administration of a therapeutic agent may reduce one or more side effects or toxicity otherwise associated with systemic administration of a therapeutic agent. In one embodiment, a therapeutic agent (eg, STING or TLR) can be conjugated to a tumor-binding antibody (eg, an antibody that binds to HER2) that delivers the therapeutic agent to a HER-2 expressing cell.

When administered in combination, the first agent, an additional agent (e.g., a second or third agent), or all of the agents may be administered in an amount or dose that is higher, lower, or the same as the amount or dose of each agent applied individually, for example, as monotherapy. In some embodiments, the administered amount or dose of the first agent, additional agent (e.g., second or third agent), or all agents is lower (e.g., at least 20%, at least 30%, at least 40%, or less than 50%) than the amount or dose of each agent used individually, for example, as monotherapy. In other embodiments, the amount or dose of the first agent, additional agent (e.g., second or third agent), or all of the agents that results in the desired effect (e.g., cancer treatment) is lower (e.g., at least 20%, according to at least 30%, at least 40% or at least 50% lower).

In some embodiments, the combinations may be in the form of an antibody molecule, e.g., a bi- or trispecific molecule, against one or more therapeutic agents selected from an antigen-presenting combination, an effector cell combination, or an anti-tumor immunosuppression combination, or any combination thereof. For example, a bispecific molecule against two or more checkpoint inhibitors (eg, an anti-PD-1 and anti-LAG-3 antibody molecule). In other embodiments, the combinations may be in the form of an antibody molecule, e.g., a bi- or tri-specific molecule, against one or more therapeutic agents selected from two or all of an antigen-presenting combination, an effector cell combination, and/or a tumor immunosuppression combination. In one embodiment, the antibody molecule is a full-length antibody or fragment thereof (eg, Fab, F(ab') ₂ , Fv, or single chain Fv fragment (scFv)). In other embodiments, the antibody molecule has a heavy chain constant region (Fc) selected from, for example, IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE heavy chain constant regions; in particular selected from, for example, IgG1, IgG2, IgG3 and IgG4 heavy chain constant regions, more specifically IgG1 or IgG4 heavy chain constant regions (eg human IgG1 or IgG4). In one embodiment, the heavy chain constant region is human IgG1 or human IgG4. In one embodiment, the constant region is altered, e.g., mutated, to change properties of the antibody molecule (e.g., to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, number of cysteine residues, effector function, or complement function). In some embodiments, the antibody molecule is in the form of a bispecific or multispecific antibody molecule, such as a bispecific, trispecific antibody molecule, as described herein.

Some examples of therapeutic agents and their combinations are presented in this application below. A more detailed description of therapeutic agents used in combinations is provided in the Detailed Description.

Immunomodulators

In some embodiments, the immunomodulator used in the combinations disclosed herein (eg, in combination with a therapeutic agent selected from an antigen-presenting combination) is an inhibitor of an immune checkpoint molecule. In one embodiment, the immunomodulator is an inhibitor of PD-1, PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, CEACAM (e.g. CEACAM-1, -3 and/or -5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGF-beta. In one embodiment, the immune checkpoint molecule inhibitor inhibits PD-1, PD-L1, LAG-3, TIM-3, CEACAM (e.g., CEACAM-1, -3 and/or -5), CTLA-4, or any combination thereof. .

Inhibition of an inhibitory molecule can be performed at the DNA, RNA or protein level. In embodiments, an inhibitory nucleic acid (eg, dsRNA, siRNA, or shRNA) can be used to inhibit the expression of an inhibitory molecule. In other embodiments, the inhibitor of the inhibitory signal is a polypeptide, such as a soluble ligand (eg, PD-1-Ig or CTLA-4 Ig) or an antibody or antigen-binding fragment thereof, that binds to an inhibitory molecule; for example, an antibody or fragment thereof (also referred to in this application as an "antibody molecule") that binds to PD-1, PD-L1, PD-L2, CEACAM (for example, CEACAM-1, -3 and/or -5) , CTLA-4, TIM-3, LAG-3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGF-beta, or a combination thereof.

In some embodiments, the antibody molecule is in the form of a bispecific or multispecific antibody molecule. In one embodiment, the bispecific antibody molecule has a first binding specificity for PD-1 or PD-L1 and a second binding specificity, e.g., a second binding specificity for TIM-3, CEACAM (e.g., CEACAM-1, -3 and/or -5 ), LAG-3 or PD-L2. In one embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and TIM-3. In another embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and LAG-3. In another embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and CEACAM (eg CEACAM-1, -3 and/or -5). In another embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and CEACAM-1. In another embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and CEACAM-3. In yet another embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1 and CEACAM-5. In another embodiment, the bispecific antibody molecule binds to PD-1 or PD-L1. In yet another embodiment, the bispecific antibody molecule binds to PD-1 and PD-L2. In another embodiment, the bispecific antibody molecule binds to TIM-3 and LAG-3. In another embodiment, the bispecific antibody molecule binds to CEACAM (eg, CEACAM-1, -3 and/or -5) and LAG-3. In another embodiment, the bispecific antibody molecule binds to CEACAM (eg, CEACAM-1, -3 and/or -5) and TIM-3. Any combination of the above molecules can be produced in a multispecific antibody molecule, e.g. a trispecific antibody, which includes a first binding specificity for PD-1 or PD-1 and a second and third binding specificity for two or more of: TIM-3, CEACAM (eg CEACAM-1, -3 and/or -5), LAG-3 or PD-L2.

In some embodiments, the implementation of the immunomodulator is an inhibitor of PD-1, such as human PD-1 (eg, an antibody molecule, as described in this application). In another embodiment, the immunomodulator is an inhibitor of PD-L1, eg human PD-L1. In one embodiment, the PD-1 or PD-L1 inhibitor is an anti-PD-1 or PD-L1 antibody molecule. A PD-1 or PD-L1 inhibitor may be administered alone or in combination with other immunomodulators, e.g. in combination with a LAG-3, TIM-3, CEACAM (e.g. CEACAM-1, -3 and/or -5) or CTLA inhibitor -four. In an exemplary embodiment, a PD-1 or PD-L1 inhibitor, eg an anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a LAG-3 inhibitor, eg an anti-LAG-3 antibody molecule. In another embodiment, a PD-1 or PD-L1 inhibitor, eg, an anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a TIM-3 inhibitor, eg, an anti-TIM-3 antibody molecule. In another embodiment, a PD-1 or PD-L1 inhibitor, e.g., an anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a CEACAM inhibitor (e.g., CEACAM-1, -3 and/or -5 inhibitors), e.g. , an anti-CEACAM antibody molecule. In another embodiment, a PD-1 or PD-L1 inhibitor, eg, an anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a CEACAM-1 inhibitor, eg, an anti-CEACAM-1 antibody molecule. In another embodiment, a PD-1 or PD-L1 inhibitor, eg, an anti-PD-1 or PD-L1 antibody molecule, is administered in combination with a CEACAM-5 inhibitor, eg, an anti-CEACAM-5 antibody molecule. In other embodiments, a PD-1 or PD-L1 inhibitor, e.g., an anti-PD-1 antibody molecule, is administered in combination with a LAG-3 inhibitor, e.g., an anti-LAG-3 antibody molecule, and a TIM-3 inhibitor, e.g., an anti-LAG-3 antibody molecule. against TIM-3. Other combinations of immunomodulators with a PD-1 inhibitor (eg, one or more of PD-L2, CTLA-4, TIM-3, LAG-3, CEACAM (eg, CEACAM-1, -3 and/or -5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGF-beta) are also included within the scope of the present invention. Any of the antibody molecules known in the art or disclosed in this application can be used in the above combinations of inhibitors of the checkpoint molecule.

In other embodiments, the immunomodulator is an inhibitor of CEACAM (eg CEACAM-1, -3 and/or -5), eg human CEACAM (eg CEACAM-1, -3 and/or -5). In one embodiment, the immunomodulator is an inhibitor of CEACAM-1, eg human CEACAM-1. In another embodiment, the immunomodulator is an inhibitor of CEACAM-3, eg human CEACAM-3. In another embodiment, the immunomodulator is an inhibitor of CEACAM-5, eg human CEACAM-5. In one embodiment, the CEACAM inhibitor (eg CEACAM-1, -3 and/or -5) is an anti-CEACAM antibody molecule (eg CEACAM-1, -3 and/or -5). A CEACAM inhibitor (eg, CEACAM-1, -3, and/or -5) may be administered alone or in combination with other immunomodulators, eg, in combination with a LAG-3, TIM-3, PD-1, PD-L1, or CTLA inhibitor -four.

In other embodiments, the immunomodulator is an inhibitor of LAG-3, such as human LAG-3. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibody molecule. The LAG-3 inhibitor can be administered alone or in combination with other immunomodulators, for example in combination with a CEACAM inhibitor (eg CEACAM-1, -3 and/or -5), TIM-3, PD-1, PD-L1 or CTLA -four.

In other embodiments, the immunomodulator is an inhibitor of TIM-3, such as human TIM-3. In one embodiment, the TIM-3 inhibitor is an anti-TIM-3 antibody molecule. The TIM-3 inhibitor can be administered alone or in combination with other immunomodulators, for example in combination with a CEACAM inhibitor (eg CEACAM-1, -3 and/or -5), LAG-3, PD-1, PD-L1 or CTLA -four.

In some embodiments, the immunomodulator used in the combinations disclosed herein (eg, in combination with a therapeutic agent selected from an antigen-presenting combination) is an activator or agonist of a costimulatory molecule. In one embodiment, the costimulatory molecule agonist is selected from an agonist (e.g., agonist antibody or antigen-binding fragment or soluble fusion construct thereof) OX40, CD2, CD27, CD28, CD, 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.

In other embodiments, the immunomodulator is a GITR agonist. In one embodiment, the GITR agonist is an anti-GITR antibody molecule. The GITR agonist may be administered alone or in combination with other immunomodulators, e.g., in combination with a PD-1 inhibitor, PD-L1, CTLA-4, CEACAM (e.g. CEACAM-1, -3 and/or -5), TIM-3 or LAG-3. In some embodiments, an anti-GITR antibody molecule is a bispecific antibody that binds to GITR and PD-1, PD-L1, CTLA-4, CEACAM (e.g., CEACAM-1, -3 and/or -5), TIM-3, or LAG-3. In one embodiment, an anti-GITR antibody molecule is administered in combination with an anti-PD-1 antibody molecule (eg, an anti-PD-1 molecule as described herein). The anti-GITR antibody molecule and the anti-PD-1 antibody molecule can be presented as a single antibody composition or as a bispecific antibody molecule. In other embodiments, a GITR agonist may be administered in combination with another costimulatory molecule, e.g., an agonist for OX40, CD2, CD27, CD28, CD, 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.

In other embodiments, the immunomodulator is an activator of a costimulatory molecule (eg, an OX40 agonist). In one embodiment, the OX40 agonist is an anti-OX40 antibody molecule. The OX40 agonist may be administered alone or in combination with other immunomodulators, e.g., in combination with a PD-1 inhibitor, PD-L1, CTLA-4, CEACAM (e.g. CEACAM-1, -3 and/or -5), TIM-3 or LAG-3. In some embodiments, the OX40 antibody molecule is a bispecific antibody that binds to GITR and PD-1, PD-L1, CTLA-4, CEACAM (e.g., CEACAM-1, -3 and/or -5), TIM-3, or LAG-3. In one embodiment, an OX40 antibody molecule is administered in combination with an anti-PD-1 antibody molecule (eg, an anti-PD-1 molecule as described herein). The OX40 antibody molecule and the anti-PD-1 antibody molecule can be presented as a single antibody composition or as a bispecific antibody molecule. In other embodiments, an OX40 agonist may be administered in combination with another co-stimulatory molecule, e.g., a GITR agonist, CD2, CD27, CD28, CD, 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.

It should be noted that only exemplary combinations of checkpoint inhibitors or co-stimulatory molecule agonists are provided herein. Additional combinations of such agents are included within the scope of the present invention.

Molecules of antibodies to PD-1

In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody molecule as described in U.S. Patent Application Publication 2015/0210769 (USSN 14/604,415) entitled Antibody Molecules to PD-1 and Uses Thereof"), incorporated in its entirety by reference. In one embodiment, an anti-PD-1 antibody molecule comprises at least one antigen-binding region, e.g., a variable region or an appropriate antigen-binding fragment, from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 -hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, three, or four variable regions from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049- hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one or two heavy chain variable regions from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one or two light chain variable regions from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, the anti-PD-1 antibody molecule comprises an IgG4 heavy chain constant region, eg, human IgG4. In one embodiment, human IgG4 includes a substitution at position 228 according to EU numbering (eg, substitution Ser for Pro). In another embodiment, the anti-PD-1 antibody molecule comprises an IgG1 heavy chain constant region, eg, human IgG1. In one embodiment, the human IgG1 includes a substitution at position 297 according to EU numbering (eg, substitution of Asn for Ala). In one embodiment, human IgG1 comprises a substitution at position 265 according to the EU numbering, a substitution at position 329 according to the EU numbering, or both of such substitutions (for example, a substitution of Asp to Ala at position 265 and a substitution of Pro to Ala at position 329). In one embodiment, human IgG1 comprises a substitution at position 234 according to EU numbering, a substitution at position 235 according to EU numbering, or both (eg, a Leu to Ala substitution at position 234 and a Leu to Ala substitution at position 235). In one embodiment, the heavy chain constant region comprises the amino acid sequence shown in Table 3, or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98 %, 99% or higher) to her.

In yet another embodiment, the anti-PD-1 antibody molecule comprises a light chain kappa constant region, eg, a human light chain kappa constant region. In one embodiment, the light chain constant region comprises the amino acid sequence shown in Table 3, or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98 %, 99% or higher) to her.

In another embodiment, the anti-PD-1 antibody molecule comprises an IgG4 heavy chain constant region, such as human IgG4, and a light chain kappa constant region, such as a human light chain kappa constant region, such as a heavy and light chain constant region, comprising the amino acid sequence shown in Table 3, or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to it. In one embodiment, human IgG4 includes a substitution at position 228 according to EU numbering (eg, substitution Ser for Pro). In yet another embodiment, an anti-PD-1 antibody molecule comprises an IgG1 heavy chain constant region, e.g., human IgG1, and a light chain kappa constant region, e.g., a human light chain kappa constant region, e.g., a heavy and light chain constant region, comprising an amino acid the sequence shown in Table 3, or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to it. In one embodiment, the human IgG1 includes a substitution at position 297 according to EU numbering (eg, substitution of Asn for Ala). In one embodiment, human IgG1 comprises a substitution at position 265 according to EU numbering, a substitution at position 329 according to EU numbering, or both (eg, Asp to Ala at position 265 and Pro to Ala at position 329). In one embodiment, human IgG1 comprises a substitution at position 234 according to the EU numbering, a substitution at position 235 according to the EU numbering, or both of such substitutions (eg, a change from Leu to Ala at position 234 and a change from Leu to Ala at position 235).

In another embodiment, an anti-PD-1 antibody molecule comprises a heavy chain variable domain and constant region, a light chain variable domain and constant region, or both, comprising the amino acid sequence BAP049-Clone-A, BAP049-Clone-B, BAP049- Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences. The PD-1 antibody molecule optionally includes a leader sequence from the heavy chain, light chain, or both, as shown in Table 4; or a sequence substantially identical to them.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three complementarity determining regions (CDRs) from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049- hum01 BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two or three CDR regions (or all CDR regions simultaneously) from a heavy chain variable region comprising the amino acid sequence shown in Table 1 or encoded by a nucleotide sequence, shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid the sequence shown in Table 1 or the encoded nucleotide sequence shown in Table 1.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 -hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions (or all CDR regions simultaneously) from a light chain variable region comprising the amino acid sequence shown in Table 1 or encoded by a nucleotide sequence, shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid the sequence shown in Table 1 or encoded by the nucleotide sequence shown in Table 1. In some embodiments, the anti-PD-1 antibody molecule comprises a substitution in the light chain CDR region, e.g., one or more substitutions in CDR1, CDR2, and/or CDR3 of the light chain. chains. In one embodiment, the anti-PD-1 antibody molecule comprises a substitution in the light chain CDR3 at position 102 of the light chain variable region, e.g., a change from a cysteine residue to a tyrosine residue or a cysteine residue to a serine residue, at position 102 of the light chain variable region according to Table 1 ( for example, SEQ ID NO: 16 or 24 for murine or chimeric, unmodified; or any of SEQ ID NO: 34, 42, 46, 54, 58, 62, 66, 70, 74, or 78 for a modified sequence).

In another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, three, four, five, or six CDR regions (or all CDR regions simultaneously) from a heavy and light chain variable region comprising the amino acid sequence shown in Table 1 or encoded by the nucleotide sequence shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, for example, amino acid substitutions or deletions compared to the amino acid sequence shown in Table 1 or the encoded nucleotide sequence shown in Table 1.

In one embodiment, an anti-PD-1 antibody molecule includes all six CDR regions from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049- hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or those described in Table 1 or encoded by the nucleotide sequence in Table 1, or closely related CDR regions, e.g., CDR regions that are identical or that have at least one amino acid change but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, such as conservative substitutions). In one embodiment, an anti-PD-1 antibody molecule may include any CDR region described herein. In some embodiments, the anti-PD-1 antibody molecule comprises a substitution in the light chain CDR region, for example, one or more substitutions in the light chain CDR1, CDR2, and/or CDR3. In one embodiment, the anti-PD-1 antibody molecule comprises a substitution in the light chain CDR3 at position 102 of the light chain variable region, e.g., a change from a cysteine residue to a tyrosine residue or a cysteine residue to a serine residue, at position 102 of the light chain variable region according to Table 1 ( for example, SEQ ID NO: 16 or 24 for murine or chimeric, unmodified; or any of SEQ ID NO: 34, 42, 46, 54, 58, 62, 66, 70, 74, or 78 for a modified sequence).

In another embodiment, an anti-PD-1 antibody molecule includes at least one, two, or three CDR regions according to Kabat et al. (e.g., at least one, two, or three CDR regions as defined by Kabat as shown in Table 1) from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049- hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which has at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, or three CDR regions according to Kabat et al., shown in Table 1.

In another embodiment, an anti-PD-1 antibody molecule includes at least one, two, or three CDR regions according to Kabat et al. (e.g., at least one, two, or three CDR regions as defined by Kabat as shown in Table 1) from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049- hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which has at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, or three CDR regions according to Kabat et al., shown in Table 1.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, three, four, five, or six CDR regions according to Kabat et al. (e.g., at least one, two, three, four, five, or six CDR regions as defined by Kabat as shown in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10, BAP049-hum11, BAP049-hum12, BAP049-hum13, BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which has at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, three, four, five, or six CDR regions according to Kabat et al. shown in Table 1.

In another embodiment, the anti-PD-1 antibody molecule includes all six CDR regions according to Kabat et al. (e.g., all six CDR regions as defined by Kabat as shown in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049- hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which has at least one amino acid change but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, eg, conservative substitutions) compared to all six CDR regions according to Kabat et al. shown in Table 1. In one embodiment, an anti-PD-1 antibody molecule may include any CDR region described herein.

In another embodiment, the anti-PD-1 antibody molecule comprises at least one, two, or three Chothia hypervariable loops (e.g., at least one, two, or three Chothia hypervariable loops as shown in Table 1) from the heavy chain variable region. an antibody described herein, for example an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09 , BAP049-hum10, BAP049-hum11, BAP049-hum12, BAP049-hum13, BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone -D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or at least amino acids from those hypervariable loops that contact PD-1; or which has at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, or three hypervariable loops according to Chothia et al. shown in Table 1.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three Chothia hypervariable loops (e.g., at least one, two, or three Chothia hypervariable loops as shown in Table 1) of the antibody light chain variable region. described in this application, for example, an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B BAP049-Clone-C BAP049-Clone- D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or at least amino acids from those hypervariable loops that contact PD-1; or which has at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, or three hypervariable loops according to Chothia et al. shown in Table 1.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, three, four, five, or six hypervariable loops (e.g., at least one, two, three, four, five, or six hypervariable loops as defined by Chothia , as shown in Table 1) from the heavy and light chain variable regions of an antibody described herein, for example, an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049 -hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A -Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or at least amino acids from those hypervariable loops that contact PD-1; or which has at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, three, four, five, or six hypervariable loops according to Chothia et al. shown in Table 1.

In one embodiment, an anti-PD-1 antibody molecule includes all six hypervariable loops (e.g., all six hypervariable loops as defined by Chothia as shown in Table 1) of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01 BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 -hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E, or closely related hypervariable loops, e.g. hypervariable loops that are identical or that have at least one amino acid change but no more than two, three or four changes (eg substitutions, deletions or insertions, eg conservative substitutions); or which have at least one amino acid change but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, eg, conservative substitutions) compared to all six hypervariable loops according to Chothia et al. shown in Table 1 In one embodiment, an anti-PD-1 antibody molecule may include any of the hypervariable loops described herein.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three hypervariable loops that have the same canonical structures as the corresponding hypervariable loop of an antibody described herein, e.g., an antibody selected from any of BAP049 -hum01 BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 , BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E, for example, the same canonical structures as at least loop 1 and/or loop 2 of the heavy and/or light chain variable domains of an antibody described herein. See, for example, Chothia et al., (1992) J. Mol. Biol. 227:799-817; Tomlinson et al., (1992) J. Mol. Biol. 227:776-798 on descriptions of the canonical structures of hypervariable loops. These structures can be determined by looking at the tables described in the indicated sources.

In some embodiments, an anti-PD-1 antibody molecule comprises a combination of CDR regions or hypervariable loops as defined according to Kabat et al. and Chothia et al.

In one embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions or hypervariable loops from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049 -hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 , BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E, as defined by Kabat and Chothia (e.g., at least one, two or three CDR regions or hypervariable loops as defined by Kabat and Chothia, as shown in Table 1); or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which has at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, or three CDR regions or hypervariable loops according to Kabat, and /or Chothia shown in Table 1.

For example, an anti-PD-1 antibody molecule may include a VH CDR1 according to Kabat et al. or VH hypervariable loop 1 according to Chothia et al., or combinations thereof, for example as shown in Table 1. In one embodiment, combinations of Kabat CDR region and Chothia CDR1 VH comprises the amino acid sequence GYTFTTYWMH (SEQ ID NO: 224) or essentially an amino acid sequence identical to it (eg, having at least one amino acid change, but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, eg, conservative substitutions)). The anti-PD-1 antibody molecule may further include, for example, VH CDR regions 2-3 according to Kabat et al. and VL CDR regions 1-3 according to Kabat et al., for example, as shown in Table 1. Thus, in some embodiments, framework regions are defined based on a combination of CDR regions defined according to Kabat et al. and hypervariable loops, defined according to Chothia et al. For example, an anti-PD-1 antibody molecule may include VH FR1 defined based on VH hypervariable loop 1 according to Chothia et al. and VH FR2 defined based on VH CDR regions 1-2 according to Kabat et al., for example, as shown in Table 1. An anti-PD-1 antibody molecule may further comprise, for example, VH FRs 3-4 defined based on VH CDRs 2-3 according to Kabat et al., and VL FRs 1-4 defined based on VL CDRs 1-3 according to Kabat et al.

An anti-PD-1 antibody molecule may contain any combination of CDR regions or hypervariable loops as defined by Kabat and Chothia. In one embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E, as defined by Kabat and Chothia (e.g., at least one, two or three CDR regions as defined by Kabat and Chothia, as shown in Table 1).

In one embodiment, an anti-PD-1 antibody molecule comprises:

(a) a heavy chain variable region (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 light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33;

(b) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 1; the amino acid sequence of VHCDR2 SEQ ID NO: 2; and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32;

(c) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224, the amino acid sequence of VHCDR2 SEQ ID NO: 5, and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33; or

(d) VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32.

In the combinations herein, in another embodiment, the anti-PD-1 antibody molecule comprises: (i) a heavy chain variable region (VH) comprising the VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 or SEQ ID NO: 5; and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and (ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10 or SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 11 or SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: : 32 or SEQ ID NO: 33.

In an embodiment, e.g., an embodiment including a variable region, a CDR region (e.g., a Chothia CDR or a Kabat CDR), or another sequence as defined herein, e.g., in Table 1, the antibody molecule is a monospecific antibody molecule. , a bispecific antibody molecule, or is an antibody molecule that includes an antigen-binding fragment of an antibody, such as a semi-antibody or an antigen-binding fragment of a semi-antibody. In some embodiments, the antibody molecule is a bispecific antibody molecule having a first binding specificity for PD-1 and a second binding specificity for TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3 and/or CEACAM- 5), PD-L1 or PD-L2. In one embodiment, the bispecific antibody molecule binds to PD-1 and TIM-3. In another embodiment, the bispecific antibody molecule binds to PD-1 and LAG-3. In another embodiment, the bispecific antibody molecule binds to PD-1 and CEACAM (eg, CEACAM-1, CEACAM-3 and/or CEACAM-5). In another embodiment, the bispecific antibody molecule binds to PD-1 and CEACAM-1. In yet another embodiment, the bispecific antibody molecule binds to PD-1 and CEACAM-5. In another embodiment, the bispecific antibody molecule binds to PD-1 and PD-L1. In yet another embodiment, the bispecific antibody molecule binds to PD-1 and PD-L2. Any combination of the above molecules can be produced in a multispecific antibody molecule, e.g., a trispecific antibody, which includes a first binding specificity for PD-1 and a second and third binding specificity for one or more of TIM-3, LAG-3, CEACAM (e.g. , CEACAM-1, CEACAM-3 or CEACAM-5), PD-L1 or PD-L2.

In other embodiments, an anti-PD-1 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, or CEACAM-5), PD- L1 or PD-L2. In one embodiment, the bispecific antibody molecule used in combination binds to CEACAM (eg, CEACAM-1, CEACAM-3 and/or CEACAM-5) and LAG-3. In another embodiment, the bispecific antibody molecule used in combination binds to CEACAM (eg, 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.

Combination Therapy Applications

The combinations disclosed herein may result in one or more of the following: increased antigen presentation, increased effector cell function (e.g., one or more of T cell proliferation, IFN-γ secretion, or cytolytic function), inhibition of regulatory T- cells, affecting the activity of multiple cell types such as regulatory T cell, effector T cells, and NK cells), an increase in tumor-infiltrating lymphocytes, an increase in T-cell receptor-mediated proliferation, and a decrease in immune escape of cancer cells. In one embodiment, the use of a PD-1 inhibitor in combinations inhibits, reduces, or neutralizes one or more PD-1 activities, resulting in blockade or a decrease in the immune checkpoint. Thus, such combinations can be used to treat or prevent disorders that require enhancement of the immune response in the subject.

Thus, in another aspect, a method for correcting an immune response in a subject is provided. The method includes administering to a subject a combination disclosed herein (e.g., a combination comprising a therapeutically effective amount of an anti-PD-1 antibody molecule), alone or in combination with one or more agents or procedures, resulting in modulation of the subject's immune response. In one embodiment, the antibody molecule enhances, stimulates or enhances the immune response in the subject. The subject may be a mammal, eg a primate, preferably a higher primate, eg a human (eg, a patient having or at risk of developing a disorder described herein). In one embodiment, the subject is in need of an enhanced immune response. In one embodiment, the subject has or is at risk of developing a disorder described in this application, such as cancer or an infectious disease, as described in this application. In some embodiments, the implementation of the subject has or is at risk of weakening the immune system. For example, the subject is or has received chemotherapy and/or radiation therapy. Alternatively, or in combination, the subject is or is at risk of being immunocompromised as a result of infection.

In one aspect, a method of treating (eg, one or more of reducing, inhibiting, or delaying progression) a cancer or tumor in a subject is provided. The method includes administering to the subject a combination disclosed herein (eg, a combination comprising a therapeutically effective amount of an anti-PD-1 antibody molecule).

In some embodiments, the cancer being treated with the combination includes, but is not limited to, solid tumor, hemoblastosis (eg, leukemia, lymphoma, myeloma, eg, multiple myeloma), and metastatic disease. In one embodiment, the cancer is a solid tumor. Examples of solid tumors include malignant tumors, such as sarcomas and carcinomas, such as adenocarcinomas of various organ systems, such as adenocarcinomas affecting the lung, breast, ovary, lymphoid tissue, gastrointestinal tract (eg, colon), anal canal, genitals and genitourinary tract (eg, kidney, urothelium, bladder cells, prostate), pharynx, CNS (eg, brain, nerve or glial cells), head and neck, skin (eg, melanoma), and pancreas, and adenocarcinomas which include cancers such as forms of colon cancer, rectal cancer, kidney cancer (eg, renal cell carcinoma (clear cell or non-clear cell renal cell carcinoma), liver cancer, lung cancer (eg, non-small cell lung cancer (squamous or non-squamous non-small cell lung cancer), small bowel cancer, and esophageal cancer Can be early, intermediate, late stage or metastatic cancer.

In one embodiment, the cancer is selected from lung cancer (e.g., non-small cell lung cancer (NSCLC) (e.g., NSCLC with squamous and/or non-squamous histology or adenocarcinoma NSCLC)), skin cancer (e.g., Merkel cell cancer, or melanoma (e.g., advanced melanoma)), kidney cancer (eg, kidney cancer (eg, renal cell carcinoma)), liver cancer, myeloma (eg, multiple myeloma), prostate cancer, breast cancer (eg, breast cancer that does not express one , two or no estrogen receptor, progesterone receptor, or Her2/neu, e.g., triple-negative breast cancer), colon and rectal cancer, pancreatic cancer, head and neck cancer (e.g., head and neck squamous cell carcinoma (HNSCC), rectal cancer, gastric and esophageal cancer, thyroid cancer (eg, anaplastic thyroid carcinoma), cervical cancer, neuroendocrine tumor (NET) (eg, atypical lung carcinoid tumor), lymphoproliferative disease (eg, post-transplant lymphoproliferative disease), or hematological malignancy, T-cell lymphoma, B-cell lymphoma, non-Hodgkin's lymphoma, or leukemia (eg, myeloid leukemia or lymphoid leukemia).

In another embodiment, the cancer is selected from carcinoma (eg, advanced or metastatic carcinoma), melanoma, or lung carcinoma, eg, non-small cell lung carcinoma.

In one embodiment, the cancer is lung cancer, such as non-small cell lung cancer or small cell lung cancer. In some embodiments, the non-small cell lung cancer is stage I (e.g., stages Ia or Ib), stage II (e.g., stages IIa or IIb), stage III (e.g., stages IIIa or IIIb) or stage IV non-small cell lung cancer.

In one embodiment, the cancer is melanoma, such as widespread melanoma. In one embodiment, the cancer is an advanced or inoperable melanoma that has not responded to other therapies. In other embodiments, the cancer is a BRAF-mutated melanoma (eg, a BRAF V600 mutation). In other embodiments, the combination disclosed herein (e.g., a combination comprising an anti-PD-1 antibody molecule) is administered following treatment with an anti-CTLA-4 antibody (e.g., ipilimumab), with or without a BRAF inhibitor (e.g., vemurafenib or dabrafenib) .

In another embodiment, the cancer is a hepatocarcinoma, eg, advanced hepatocarcinoma, with or without a viral infection, eg, chronic viral hepatitis.

In another embodiment, the cancer is prostate cancer, eg advanced prostate cancer.

In yet another embodiment, the cancer is a myeloma, such as multiple myeloma.

In another embodiment, the cancer is kidney cancer, eg, renal cell carcinoma (RCC) (eg, metastatic RCC, non-clear cell renal cell carcinoma (nccRCC), or clear cell renal cell carcinoma (CCRCC)).

In one embodiment, the cancer microenvironment has an elevated level of PD-L1 expression. Alternatively, or in combination, the cancer microenvironment may have increased expression of IFNγ and/or CD8.

In some embodiments, the subject has, or has been found to have, a tumor that has one or more of a high level or expression of PD-L1 and/or is a tumor infiltrating lymphocyte (TIL)+ (eg, has an increased number of TIL lymphocytes). In some embodiments, the subject has, or has been found to have, a tumor that has a high level or expression of PD-L1 and that is TIL+. In some embodiments, the methods described herein further comprise identifying a subject based on the presence of a tumor that has one or more of a high level or expression of PD-L1 and/or is TIL+. In some embodiments, the methods described herein further comprise identifying a subject based on the presence of a tumor that has a high level or expression of PD-L1 and is TIL+. In some embodiments, tumors that are TIL+ are CD8 and IFNγ positive. In some embodiments, the subject has, or has been found to have, a high percentage of cells that are positive for one, two, or more of PD-L1, CD8, and/or IFNγ. In some embodiments, the subject has, or has been found to have, a high percentage of cells that are positive for all of PD-L1, CD8, and IFNγ.

In some embodiments, the methods described herein further comprise identifying a subject based on the presence of a high percentage of cells that are positive for one, two, or more of PD-L1, CD8, and/or IFNγ. In some embodiments, the methods described herein further include identifying a subject based on the presence of a high percentage of cells that are positive for all of PD-L1, CD8, and IFNγ. In some embodiments, the subject has, or has been found to have, one, two or more of PD-L1, CD8, and/or IFNγ, and one or more of a lung cancer, eg, squamous cell lung cancer or lung adenocarcinoma (eg, NSCLC); head and neck cancer; squamous cell carcinoma of the cervix; stomach cancer; esophageal cancer; thyroid cancer (eg, anaplastic thyroid carcinoma); skin cancer (eg, Merkel cell cancer or melanoma), breast cancer (eg, TNBC), and/or nasopharyngeal cancer (NPC). In some embodiments, the methods described herein further describe identifying a subject based on the presence of one, two, or more of PD-L1, CD8, and/or IFNγ, and one or more of a lung cancer, e.g., squamous cell lung cancer or lung adenocarcinoma. (eg, NSCLC); head and neck cancer; squamous cell carcinoma of the cervix; stomach cancer; thyroid cancer (eg, anaplastic thyroid carcinoma); skin cancer (eg, Merkel cell cancer or melanoma), neuroendocrine tumor, breast cancer (eg, TNBC), and/or nasopharyngeal cancer.

The methods and compositions disclosed in this application can be used to treat metastatic lesions associated with the above forms of cancer.

In another aspect of the invention, a method of treating an infectious disease in a subject is provided, comprising administering to the subject a combination as described herein, for example, a combination comprising a therapeutically effective amount of an anti-PD-1 antibody molecule described herein. In one embodiment, the infectious disease is selected from hepatitis (eg hepatitis C) or sepsis.

In addition, the invention provides a method for enhancing an immune response to an antigen in a subject, comprising administering to the subject: (i) the antigen; and (ii) a combination as described herein, for example, a combination comprising a therapeutically effective amount of an anti-PD-1 antibody molecule described herein, resulting in an enhanced immune response to the antigen in the subject. The antigen may be, for example, a tumor antigen, a viral antigen, a bacterial antigen, or an antigen from a pathogen.

Combinations as described herein may be administered to a subject systemically (eg, orally, parenterally, subcutaneously, intravenously, rectally, intramuscularly, intraperitoneally, intranasally, transdermally, or by inhalation, or by intracavitary insertion), topically, or by application to mucous membranes, such like the nose, throat and bronchioles.

Doses and schedules for the use of therapeutic agents disclosed in this application can be determined by a specialist. In some embodiments, the anti-PD-1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose of about 1-30 mg/kg, e.g., about 5-25 mg/kg, about 10-20 mg/kg, about 1 -5 mg/kg or approximately 3 mg/kg. The schedule of application may vary, for example, from once a week to once every 2, 3 or 4 weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 10-20 mg/kg biweekly.

In some embodiments, the anti-PD-1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose (e.g., fixed dose) of about 100 mg to 600 mg, such as about 200 mg to 500 mg, such as about 250 mg to 450 mg, about 300 mg to 400 mg, about 250 mg to 350 mg, about 350 mg to 450 mg, or about 100 mg, about 200 mg, about 300 mg, or about 400 mg. The schedule of administration (eg, a fixed dose regimen) may vary, for example, from once a week to once every 2, 3, 4, 5 or 6 weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg to 400 mg once every three weeks or once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every three weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every three weeks.

In one embodiment, the anti-PD-1 antibody molecule is administered alone or in combination (eg, in combination with an anti-LAG-3 antibody molecule), at a dose of less than or about 5 mg/kg; less than or about 4 mg/kg; less than or about 3 mg/kg; less than or about 2 mg/kg; less than or approximately 1 mg/kg every other week. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of 1-5 mg/kg biweekly; 1–4 mg/kg biweekly, 1–3 mg/kg biweekly, or 1–2 mg/kg biweekly. In one embodiment, the anti-LAG-3 antibody molecule is administered alone or in combination (eg, in combination with an anti-PD-1 antibody molecule) at a dose of 1-5 mg/kg biweekly; 1–4 mg/kg biweekly, 1–3 mg/kg biweekly, or 1–2 mg/kg biweekly.

The antibody molecules described herein are preferred for use in the methods described herein, although other anti-PD-1 antibodies may be used instead or in combination with an anti-PD-1 antibody molecule of the invention.

Additional Combination Therapies

The methods and combinations described in this application may include or be used in combination with other agents or therapeutic methods. In one embodiment, the methods described herein comprise administering to a subject a combination comprising an anti-PD-1 antibody molecule as described herein, in combination with an agent or therapeutic procedure or method, in an amount effective to treat or prevent the disorder. The anti-PD-1 antibody molecule and the agent or therapeutic procedure or method may be administered simultaneously or sequentially, in any order. Any combination and sequence of anti-PD-1 antibody molecules and other therapeutic agents, procedures, or methods may be used (eg, as described herein). The antibody molecule and/or other therapeutic agents, procedures or methods may be used during periods of active disorder or during periods of remission or less active disease. The antibody molecule can be used before another treatment, concurrently with treatment, after treatment, or during remission of a disorder.

In some embodiments, the methods and compositions described herein are used in combination with one or more other antibody molecules, chemotherapy, other anticancer therapy (e.g., targeted anticancer therapies, gene therapy, viral therapy, RNA therapy, bone marrow transplantation, nanotherapy, or oncolytic agents), cytotoxic agents, immunotherapy (eg, cytokine or cell-based immunotherapy), surgery (eg, lumpectomy or mastectomy), or radiotherapy procedures, or a combination of any of the foregoing. Complementary therapy may be in the form of adjuvant or neoadjuvant therapy. In some embodiments, the complementary therapy is an enzyme inhibitor (eg, a small molecule enzyme inhibitor) or a metastatic inhibitor. Examples of cytotoxic agents that can be used in combination include antimicrotubulin agents, topoisomerase inhibitors, antimetabolites, mitosis inhibitors, alkylating agents, anthracyclines, vinca alkaloids, intercalating agents, agents capable of blocking the signal transduction pathway, agents that induce apoptosis, proteasome inhibitors, and ionizing radiation (for example, local or total exposure (for example, exposure to gamma radiation)). In other embodiments, the complementary therapy is surgery or radiation therapy, or a combination thereof. In other embodiments, the adjunctive therapy is one or more of the PI3K/AKT/mTOR pathway, an HSP90 inhibitor, or a tubulin inhibitor.

Alternatively, or in combination with the above combinations, the methods and compositions described herein may include, used in combination with one or more of: an immunomodulator (eg, an activator of a costimulatory molecule or an inhibitor of an inhibitory molecule, eg, an immune checkpoint molecule); vaccines, for example a therapeutic cancer vaccine; or other forms of cellular immunotherapy.

Exemplary non-limiting combinations and uses of the combinations disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, include the following.

In some embodiments, a combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with a co-stimulatory molecule or inhibitory molecule modulator, such as a co-inhibitory ligand or receptor.

In one embodiment, the combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with a modulator, eg an agonist, of a co-stimulatory molecule. In one embodiment, the costimulatory molecule agonist is selected from an agonist (e.g., agonist antibody or antigen-binding fragment thereof, or soluble fusion construct) OX40, CD2, CD27, CD, 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.

In one embodiment, the combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with an inhibitor of an inhibitory molecule (or immune checkpoint molecule) selected from PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, CEACAM (eg CEACAM-1, CEACAM-3 and/or CEACAM-5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGF-beta. In one embodiment, the inhibitor is a soluble ligand (eg, CTLA-4-Ig) or an antibody or antibody fragment that binds to PD-L1, PD-L2, or CTLA-4. For example, an anti-PD-1 antibody molecule can be administered in combination with an anti-CTLA-4 antibody, such as ipilimumab, for example, for the treatment of cancer (for example, a cancer selected from: melanoma, for example, metastatic melanoma; lung cancer, for example, non-small cell cancer lung; or prostate cancer). In one embodiment, an anti-PD-1 antibody molecule is administered following treatment with an anti-CTLA-4 antibody (eg, ipilimumab), with or without a BRAF inhibitor (eg, vemurafenib or dabrafenib).

In another embodiment, the combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with an anti-LAG-3 antibody or an antigen-binding fragment thereof.

In another embodiment, the combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with an anti-TIM-3 antibody or an antigen-binding fragment thereof.

In other embodiments, the combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with an anti-LAG-3 antibody and an anti-TIM-3 antibody (or antigen-binding fragments thereof).

In another embodiment, a combination disclosed herein, e.g., a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with a CEACAM inhibitor (e.g., a CEACAM-1 and/or CEACAM-5 inhibitor), e.g., an antibody molecule against CEACAM. In another embodiment, an anti-PD-1 antibody molecule is administered in combination with a CEACAM-1 inhibitor, eg, an anti-CEACAM-1 antibody molecule. In another embodiment, an anti-PD-1 antibody molecule is administered in combination with a CEACAM-5 inhibitor, eg, an anti-CEACAM-5 antibody molecule.

The combination of antibodies listed in this application can be administered separately, for example, as separate antibodies or antigen-binding fragments, or associated, for example, as a bispecific or trispecific antibody molecule. In one embodiment, a bispecific antibody is administered that comprises an anti-PD-1 antibody molecule and an anti-TIM-3, anti-CEACAM (e.g. anti-CEACAM-1, CEACAM-3 and/or anti-CEACAM-5) or anti-LAG-3 antibody, or an antigen-binding fragment thereof. In some embodiments, the combination of antibodies listed in this application is used to treat cancer, for example, cancer described in this application (eg, solid tumor or hemoblastosis).

In other embodiments, the implementation of the combination disclosed in this application, for example, a combination comprising an antibody molecule against PD-1, includes or is used in combination with a cytokine. The cytokine can be administered as a molecule fused to an anti-PD-1 antibody molecule or as separate compositions. In one embodiment, the anti-PD-1 antibody is administered in combination with one, two, three or more cytokines, eg, as a fusion molecule or as separate compositions. In one embodiment, the cytokine is an interleukin (IL) selected from one, two, three or more of IL-1, IL-2, IL-12, IL-15, or IL-21. In one embodiment, the bispecific antibody molecule has a first binding specificity for a first target (e.g., PD-1), a second binding specificity for a second target (e.g., LAG-3 or TIM-3), and optionally linked to an interleukin domain (e.g., , IL-12), for example, full-length IL-12 or part of it. In some embodiments, the combination of an anti-PD-1 antibody molecule and a cytokine described herein is used to treat a cancer, eg, a cancer described herein (eg, a solid tumor).

In some embodiments, a combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with an antibody specific for HLA C, for example, an antibody specific for killer cell immunoglobulin-like receptors (also specified in present application as "Anti-KIR antibody"). In some embodiments, the combination of an anti-PD-1 antibody molecule and an anti-KIR antibody is used to treat cancer, eg, the cancer described herein (eg, a solid tumor, eg, an advanced solid tumor).

In one embodiment, the combination disclosed in this application, for example, a combination comprising an anti-PD-1 antibody molecule, includes or is used in combination with cellular immunotherapy (for example, Provenge® (for example, Sipuleucel-T)) and optionally in combination with cyclophosphamide. In some embodiments, a combination of an anti-PD-1 antibody molecule, Provenge®, and/or cyclophosphamide is used to treat cancer, eg, the cancer described herein (eg, prostate cancer, eg, advanced prostate cancer).

In another embodiment, a combination disclosed herein, e.g., a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with a vaccine, e.g., a cancer vaccine (e.g., dendritic cell-based renal cell carcinoma (DC-RCC) )). In one embodiment, the vaccine is a peptide-based, DNA-based, RNA-based, or antigen-based vaccine, or a combination thereof. In embodiments, the vaccine comprises one or more peptides, nucleic acids (eg, DNA or RNA), antigens, or a combination thereof. In some embodiments, the combination of an anti-PD-1 antibody molecule and a DC-RCC vaccine is used to treat cancer, e.g., the cancer described herein (e.g., kidney cancer, e.g., metastatic renal cell carcinoma (RCC) or clear cell renal cell carcinoma). carcinomas (CCRCC)).

In another embodiment, the combination disclosed in this application, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with an adjuvant.

In yet another embodiment, the combination disclosed in this application, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with chemotherapy and/or immunotherapy. For example, an anti-PD-1 antibody molecule can be used to treat myeloma, alone or in combination with one or more of: chemotherapy or other anticancer agents (e.g., thalidomide analogs, e.g., lenalidomide), anti-TIM-3 antibodies, tumor antigen-determined dendritic cells, tumor cells fused (eg, by electrofusion) with dendritic cells, or vaccinated with an idiotype immunoglobulin produced by malignant plasma cells. In one embodiment, an anti-PD-1 antibody molecule is used in combination with an anti-TIM-3 antibody for the treatment of myeloma, eg, multiple myeloma.

In one embodiment, a combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with chemotherapy for the treatment of lung cancer, eg, non-small cell lung cancer. In one embodiment, the anti-PD-1 antibody molecule is used with standard lung cancer, eg, NSCLC, chemotherapy, eg, platinum derivative dual therapy for the treatment of lung cancer. In other embodiments, the anti-PD-1 antibody molecule is used in combination with an indolamine-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), indoxymod (1-methyl-D-tryptophan), α-cyclohexyl-5H-imidazo[5,1-a]isoindole-5-ethanol (also known as NLG919), etc.) in a subject with advanced or metastatic cancer (eg, a patient with metastatic and recurrent NSCLC).

In other embodiments, a combination disclosed herein, e.g., a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with one or more of the following: an immune strategy (e.g., interleukin-2 or interferon-α), a targeting agent actions (eg, an inhibitor of VEGF, such as a monoclonal antibody to VEGF); a VEGF tyrosine kinase inhibitor such as sunitinib, sorafenib, axitinib and pazopanib; RNAi inhibitor; or an inhibitor of the VEGF signaling mediator, eg, a mammalian target of rapamycin (mTOR), eg, everolimus and temsirolimus. Any of these combinations may be used to treat kidney cancer, e.g., renal cell carcinoma (RCC) (e.g., clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC), or metastatic RCC, or liver cancer (e.g. , hepatocellular carcinoma).

In some embodiments, a combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with a MEK inhibitor (eg, a MEK inhibitor as described herein). In some embodiments, the combination of an anti-PD-1 antibody and a MEK inhibitor is used to treat cancer (eg, the cancer described herein). In some embodiments, the cancer being treated with the combination is selected from melanoma, colon and rectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, hemoblastosis, or renal cell carcinoma. In some embodiments, the cancer comprises a BRAF mutation (eg, a BRAF V600E mutation), wild-type BRAF, wild-type KRAS, or an activating KRAS mutation. Cancer can be early, intermediate, or advanced.

In another embodiment, the combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used in combination with one, two, or all of oxaliplatin, leucovorin, or 5-FU (eg, FOLFOX combination therapy). Alternatively, or in combination, the combination further comprises a VEGF inhibitor (eg, a VEGF inhibitor as disclosed herein). In some embodiments, a combination of an anti-PD-1 antibody, FOLFOX combination therapy, and a VEGF inhibitor is used to treat cancer (eg, the cancer described herein). In some embodiments, the cancer being treated with the combination is selected from melanoma, colon and rectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, hemoblastosis, or renal cell carcinoma. Cancer can be early, intermediate, or advanced.

In other embodiments, the combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, comprises or is used with a tyrosine kinase inhibitor (eg, axitinib) for the treatment of renal cell carcinoma and other solid tumors.

In other embodiments, a combination disclosed herein, e.g., a combination comprising an anti-PD-1 antibody molecule, comprises or is administered with an agent that targets the 4-1BB receptor (e.g., an antibody that stimulates 4-1BB signaling (CD -137), such as PF-2566). In one embodiment, an anti-PD-1 antibody molecule is administered in combination with a tyrosine kinase inhibitor (eg, axitinib) and an agent targeting the 4-1BB receptor.

An anti-PD-1 antibody molecule may be associated with a substance, such as a cytotoxic agent or molecule (eg, a therapeutic drug; a radiation-emitting compound; molecules of plant, fungal, or bacterial origin; or a biological protein (eg, a protein toxin), or a particle (eg, a recombinant viral particle, eg, through a viral envelope protein.) For example, an antibody may be coupled to a radioactive isotope such as an α-, β- or γ-emitter or a β- and γ-emitter.

Any combination and sequence of anti-PD-1 antibody molecules and other therapeutic agents, procedures, or techniques may be used (eg, as described herein). The antibody molecule and/or other therapeutic agents, procedures or techniques may be used during periods of active disorder or during periods of remission or less active disease. The antibody molecule can be used before another treatment, concurrently with treatment, after treatment, or during remission of a disorder.

Additional Combination Therapies

In some embodiments, any of the combinations disclosed herein, alternatively or in combination, further includes one or more of the agents described in Table 7.

In some embodiments, the additional therapeutic agent is selected from one or more of the following: 1) a protein kinase C (PKC) inhibitor; 2) inhibitor of heat shock protein 90 (HSP90); 3) an inhibitor of phosphoinositide 3-kinase (PI3K) and/or a target of rapamycin (mTOR); 4) a cytochrome P450 inhibitor (eg, a CYP17 inhibitor or a 17-alpha-hydroxylase/C17-20 lyase inhibitor); 5) a substance that forms complex compounds with iron; 6) an aromatase inhibitor; 7) a p53 inhibitor, eg an inhibitor of the p53/Mdm2 interaction; 8) apoptosis inducer; 9) an angiogenesis inhibitor; 10) aldosterone synthase inhibitor; 11) receptor smoothened inhibitor (SMO); 12) prolactin receptor (PRLR) inhibitor; 13) Wnt signal inhibitor; 14) CDK4/6 inhibitor; 15) 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 (eg FLK2/STK1) or PKC; 18) an inhibitor of one or more of VEGFR-2 (eg, FLK-1/KDR), PDGFR-beta, c-KIT, or Raf kinase C; 19) an inhibitor of somatostatin agonist and/or growth hormone release; 20) an anaplastic lymphoma kinase (ALK) inhibitor; 21) inhibitor of insulin-like growth factor receptor 1 (IGF-1R); 22) inhibitor of P-glycoprotein 1; 23) vascular endothelial growth factor receptor (VEGFR) inhibitor; 24) BCR-ABL kinase inhibitor; 25) FGFR inhibitor; 26) CYP11B2 inhibitor; 27) an HDM2 inhibitor, eg an HDM2-p53 interaction inhibitor; 28) tyrosine kinase inhibitor; 29) c-MET inhibitor; 30) JAK inhibitor; 31) DAC inhibitor; 32) 11β-hydroxylase inhibitor; 33) IAP inhibitor; 34) PIM kinase inhibitor; 35) porcupine inhibitor; 36) a BRAF inhibitor, eg BRAF V600E or wild-type BRAF; 37) HER3 inhibitor; 38) MEK inhibitor; or 39) a lipid kinase inhibitor, for example, as described herein and in Table 7.

In one embodiment, the additional therapeutic agent is selected from one or more of the following: Compound A8, Compound A17, Compound A23, Compound A24, Compound A27, Compound A29, Compound A33, and Compound A13.

In other embodiments, the additional therapeutic agent is selected from one or more of the following: Compound A5, Compound A8, Compound A17, Compound A23, Compound A24, Compound A29, and Compound A40.

In other embodiments, the additional therapeutic agent is selected from one or more of the following: Compound A9, Compound A16, Compound A17, Compound A21, Compound A22, Compound A25, Compound A28, Compound A48, and Compound 49.

In one embodiment, the cancer is selected from lung cancer (e.g., non-small cell lung cancer (NSCLC) (e.g., NSCLC with squamous and/or non-squamous histology or adenocarcinoma NSCLC), or disclosed in the publication listed in Table 7.

Biomarkers

In another aspect, the present application provides a method for evaluating or monitoring the effectiveness of a therapy (eg, combination therapy) described herein in a subject (eg, a subject having cancer, eg, the cancer described in this application). The method includes registering a therapy effectiveness value, where said value indicates the therapy effectiveness.

In embodiments, the treatment efficacy value includes a score of one, two, three, four, five, six, seven, eight, nine, or more (e.g., all) of the following:

(i) a tumor infiltrating lymphocyte (TIL) phenotype parameter;

(ii) a myeloid cell population parameter;

(iii) a surface marker expression parameter;

(iv) an immunological response biomarker parameter;

(v) a systemic cytokine modulation parameter;

(vi) free circulating DNA (scDNA) parameter;

(vii) systemic immunomodulation parameter;

(viii) microbiome parameter;

(ix) an activation marker parameter in a circulating immune cell; or

(x) circulating cytokine parameter.

In some embodiments, the TIL phenotype parameter includes the level or activity of one, two, three, four, or more (e.g., all) of hematoxylin and eosin (HE) staining per TIL, CD8, FOXP3, CD4, or CD3 count, in a subject, e.g., in a subject sample (eg, a tumor sample).

In some embodiments, the myeloid cell population parameter includes the level or activity of one or both of CD68 or CD163 in a subject, eg, in a sample of the subject (eg, a tumor sample).

In some embodiments, a surface marker expression parameter includes the level or activity of one, two, or more (e.g., all) of PD-L1, LAG-3, or TIM-3 in a subject, e.g., in a sample of the subject (e.g., a tumor sample).

In some embodiments, the immunological response biomarker parameter includes the level or sequence of one or more nucleic acid-based markers in a subject, eg, in a sample of the subject (eg, a tumor sample).

In some embodiments, the systemic cytokine modulation parameter includes the level or activity of one, two, three, four, five, six, seven, eight, or more (e.g., all) of IL-18, IFN-γ, ITAC (CXCL11), IL- 6, IL-10, IL-4, IL-17, IL-15, or TGF-beta in a subject, eg, in a sample of the subject (eg, a blood sample, eg, a blood plasma sample).

In some embodiments, the fsDNA parameter includes the sequence or level of one or more circulating tumor DNA (scDNA) molecules in a subject, eg, in a sample of the subject (eg, a blood sample, eg, a blood plasma sample).

In some embodiments, the systemic immunomodulation parameter includes a phenotypic characteristic of an activated immune cell, e.g., a CD3 expressing cell, a CD8 expressing cell, or both, in a subject, e.g., in a subject sample (e.g., a blood sample, e.g., a PBMC sample).

In some embodiments, the microbiome parameter includes the sequence or expression level of one or more genes in the microbiome of a subject, eg, in a sample of the subject (eg, a stool sample).

In some embodiments, an activation marker parameter in a circulating immune cell includes the level or activity of one, two, three, four, five, or more (e.g., all) of circulating CD8+, HLA-DR+Ki67+, T cells, IFN-γ, IL -18 or CXCL11 (IFN-γ induced CCK) expressing cells in a sample (eg a blood sample, eg a blood plasma sample).

In some embodiments, the circulating cytokine parameter includes the level or activity of IL-6 in the subject, eg, in a sample of the subject (eg, a blood sample, eg, a blood plasma sample).

In some embodiments of any of the methods disclosed in this application, therapy includes a combination of:

(a) an inhibitor of an immune checkpoint molecule, such as a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule); and

(b) one or more (for example, two, three, four or more) of the following:

(i) IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2 -oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Pat. No. 8,552,003;

(ii) an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318;

(iii) a DAC inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO 2014/072493;

(iv) an IL-1β inhibitor, canakinumab, or a compound disclosed in PCT publication WO 2002/16436;

(v) an IL-17 inhibitor, CJM112, or a compound disclosed in PCT publication WO 2014/122613;

(vi) a MEK inhibitor or trametinib; or

(vii) EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazole -2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT Publication WO 2013/184757.

In some embodiments of any of the methods disclosed in this application, a quantitative indicator of one or more of (i)-(x) is obtained from a sample obtained from the subject. In some embodiments, the sample is selected from a tumor sample, a blood sample (eg, a plasma sample or a PBMC sample), or a stool sample.

In some embodiments of any of the methods disclosed herein, the subject is evaluated before, during, or after receiving therapy, such as combination therapy.

In some embodiments of any of the methods disclosed herein, the quantitative index of one or more of (i)-(x) evaluates the profile for one or more of gene expression, flow cytometry or protein expression.

In some embodiments of any of the methods disclosed herein, the presence of an increased level or activity of one, two, three, four, five, or more (e.g., all) of circulating CD8+, HLA-DR+Ki67+, T cells, IFN- γ, IL-18 or CXCL11 (IFN-γ induced CCK) expressing cells and/or the presence of a reduced level or activity of IL-6 in a subject or sample is a positive predictive indicator of the effectiveness of therapy.

Alternatively, or in combination with the value sensitive methods disclosed herein, one, two, three, four or more (e.g., all) of the following are performed:

(i) prescribing a therapy (eg, combination therapy) to the subject;

(ii) administering a variable dose of therapy (eg combination therapy);

(iii) changing the regimen or time period of therapy (eg combination therapy);

(iv) administering to the subject an additional agent (eg, a therapeutic agent described herein) in combination with a therapy (eg, combination therapy); or

(v) the appointment of an alternative therapy to the subject (for example, the therapy described in this application).

Additional Embodiments

In additional embodiments, a method of treating cancer is provided, comprising: detecting in a subject or sample (e.g., a subject sample including cancer cells and optionally immunocytes such as TIL) the presence of one, two, or all of PD-L1, CD8, or IFN -γ, yielding a value for one, two, or all of PD-L1, CD8, and IFN-γ. The method may further include comparing the PD-L1, CD8, and/or IFN-γ values with a reference value, eg, a control value. If the PD-L1, CD8 and/or IFN-γ value is greater than a reference value, such as a control value, the subject is administered a therapeutically effective amount of the combination described herein (for example, a combination that includes an anti-PD-1 antibody described herein). ), optionally in combination with one or more other agents, resulting in cancer treatment. The cancer may be, for example, the cancer described in this application, such as lung cancer (squamous cell), lung cancer (adenocarcinoma), head and neck cancer, cervical cancer (squamous cell), gastric cancer, thyroid cancer (e.g. anaplastic carcinoma thyroid), skin cancer (eg, Merkel cell carcinoma or melanoma), nasopharyngeal cancer, neuroendocrine tumor (eg, atypical pulmonary carcinoid tumour), or breast cancer, eg, TN breast cancer, eg, IM-TN breast cancer . In some embodiments, the cancer is ER+ breast cancer or pancreatic cancer.

Also provided is a method for treating cancer, comprising: examining a subject or sample (e.g., a sample of a subject including cancer cells) for the presence of PD-L1, determining a PD-L1 value, comparing the PD-L1 value with a control value, and if the PD-L1 value is L1 exceeds the control value, administration to the subject of a therapeutically effective amount of the combination described in this application (for example, a combination that includes an antibody against PD-1 described in this application), optionally in combination with one or more other agents, resulting in treatment cancer. The cancer may, for example, be a cancer described herein, such as non-small cell (NSCLC) adenocarcinoma (ACA), NSCLC squamous cell carcinoma (SCC), or hepatocellular carcinoma (HCC).

In another aspect of the invention, diagnostic or therapeutic kits are provided that include the antibody molecules described herein and instructions for use.

All publications, patent applications, patents and other references cited in this application are incorporated by reference in their entirety.

Other features, objects and advantages of the invention will become apparent from the description and drawings, as well as from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows the amino acid sequences of the light and heavy chain variable regions of the mouse anti-PD-1 mAb, BAP049. Upper and lower sequences are from two independent analyses. The sequences of the light and heavy chain CDR regions based on Kabat numbering are underlined. The sequences of the light and heavy chain CDR regions based on Chothia numbering are in bold italics. The unpaired Cys residue at position 102 of the light chain sequence is boxed. The sequences are disclosed in SEQ ID NOs: 8, 228, 16 and 229, respectively, in order of appearance.

Figure 2A shows the amino acid sequences of the light and heavy chain variable regions of the mouse (Mu) mAb against PD-1, BAP049, aligned with the germline sequences. The top and bottom sequences are the germline (GL) and BAP049 (Mu mAb) sequences, respectively. The sequences of the light and heavy chain CDR regions based on Kabat numbering are underlined. The sequences of the light and heavy chain CDR regions based on Chothia numbering are in bold italics. "-" means identical amino acid residue. The sequences are disclosed in SEQ ID NOs: 230, 8, 231 and 16, respectively, in order of appearance.

Figure 2B shows the mouse κ J2 gene sequence and corresponding mutation in the mouse anti-PD-1 mAb, BAP049. "-" means identical nucleotide residue. The sequences are disclosed in SEQ ID NOs: 233, 232, 234 and 235, respectively, in order of appearance.

Figures 3A-3B show competitive binding between a fluorescently labeled mouse anti-PD-1 mAb, BAP049 (Mu mAb), and three chimeric variants of BAP049 (Chi mAb). The experiment was carried out twice and the results are shown in Figures 3A and 3B, respectively. The three BAP049 chimeric antibodies (Chi mAb (Cys), Chi mAb (Tyr), and Chi mAb (Ser)) have a Cys, Tyr, and Ser residue at position 102 of the light chain variable region, respectively. Chi mAb (Cys), Chi mAb (Tyr), and Chi mAb (Ser) are also known as BAP049-chi, BAP049-chi-Y, and BAP049-chi-S, respectively.

Figure 4 is a bar graph showing the results of a FACS binding assay for sixteen humanized clones of BAP049 (BAP049-hum01 - BAP049-hum16). Antibody concentrations are 200, 100, 50, 25, and 12.5 ng/mL from left column to right column for each mAb tested.

Figure 5 shows the structural analysis of humanized clones of BAP049 (a, b, c, d and e denote different types of framework sequences). The mAb concentrations in the samples are also shown.

Figures 6A-6B show the binding affinity and specificity of humanized BAP049 mAbs measured in a competitive binding assay using a constant concentration of Alexa 488-labeled mouse BAP049 mAb, serial dilutions of test antibodies, and 300.19 PD-1 expressing cells. The experiment was carried out twice and the results are shown in Figures 6A and 6B, respectively.

Figure 7 shows the ranking of humanized clones of BAP049 according to FACS, competitive binding analysis and structural analysis. The mAb concentrations in the samples are also shown.

Figures 8A-8B show blocking of ligand binding to PD-1 in the presence of selected clones of humanized BAP049. Blocking the binding of PD-L1-Ig and PD-L2-Ig to PD-1 is shown in Figure 8A. Blocking the binding of PD-L2-Ig to PD-1 is shown in Figure 8B. BAP049-hum01, BAP049-hum05, BAP049-hum08, BAP049-hum09, BAP049-hum10 and BAP049-hum11 were evaluated. Also included in the assays are the mouse mAb BAP049 and the chimeric mAb containing Tyr at position 102 of the light chain variable region.

Figures 9A-9B show the sequence alignment of the heavy chain variable domain for sixteen clones of humanized BAP049 and chimeric BAP049 (BAP049-chi). Figure 9A shows all sequences (SEQ ID NOs: 22, 38, 38, 38, 38, 38, 38, 38, 38, 38, 50, 50, 50, 50, 82, 82 and 86, respectively, in order of appearance ). Figure 9B shows only those amino acid sequences that differ from the mouse sequence (SEQ ID NOs: 22, 38, 38, 38, 38, 38, 38, 38, 38, 38, 50, 50, 50, 50, 82, 82 and 86, respectively, in order of appearance).

Figures 10A-10B show the sequence alignment of the light chain variable domain for sixteen clones of humanized BAP049 and chimeric BAP049 (BAP049-chi). Figure 10A shows all sequences (SEQ ID NOs: 24, 66, 66, 66, 66, 70, 70, 70, 58, 62, 78, 74, 46, 46, 42, 54 and 54, respectively, in order of appearance ). Figure 10B shows only those amino acid sequences that differ from the mouse sequence (SEQ ID NOs: 24, 66, 66, 66, 66, 70, 70, 70, 58, 62, 78, 74, 46, 46, 42, 54 and 54, respectively, in order of appearance).

Figure 11 is a schematic diagram describing antigen processing and presentation, effector cell responses, and immunosuppression pathways targeted by the combination therapy disclosed in this application.

Figure 12 shows the calculated concentrations of C _trough (C _min ) for patients of different weights receiving the same dose of an exemplary anti-PD-1 antibody molecule. Comparison of the calculated average stationary residual concentration of C _trough after administration of a constant/fixed dose depending on body weight in two schemes for the administration of an anti-PD-1 antibody molecule is shown.

Figure 13 shows observed and predicted based on the model (in the aggregate or in individual patients) concentrations of Cmin.

Figures 14A-14B show the accumulation of data, temporal dynamics and intrasubject variability of the model used to study the pharmacokinetics. The shaded areas correspond to the 90% predicted interval; solid lines correspond to the median of the prediction at each point; black dots correspond to the observed pharmacokinetic data.

Figure 15 shows the percentage of mice that survived after implantation of MC38 cells and administration of an anti-PD-1 antibody, Compound A21, or a combination of an anti-PD-1 antibody and Compound A21.

Figure 16A shows the body weight of ten animals in the group treated with vehicle, anti-mouse PD-1 antibody (four doses of 10 mg/kg, iv, once a week), panobinostat (Compound A19, five doses of 10 mg/kg, times in two days) or a combination of panobinostat and antibodies against mouse PD-1.

Figure 16B shows individual tumor responses to treatment with vehicle control, anti-mouse PD-1 antibody, panobinostat (Compound A19), or a combination of panobinostat and anti-mouse PD-1 antibody.

Figure 17 shows the percentage of surviving A20 lymphoma allograft-bearing mice following administration of Compound A40, ibrutinib, anti-PD-L1 antibody, combination of Compound A40 and anti-PD-L1 antibody, or combination of ibrutinib and anti-PD-L1 antibody (anti-PD-L1 antibody designated like aPD-L1).

Figure 18 shows mean tumor volume in A20 lymphoma allograft-bearing mice following administration of Compound A40, ibrutinib, anti-PD-L1 antibody, combination of Compound A40 and anti-PD-L1 antibody, or combination of ibrutinib and anti-PD-L1 antibody (anti-PD-L1 antibody). L1 is designated as aPD-L1).

Figure 19 shows a phase I/II design of first-time human use of an exemplary anti-PD-1 antibody molecule in patients with advanced solid tumors. *At least 21 patients were required to determine the MTD. Tumor scores were performed at screening, then every 2 courses (1 course=28 days) ±1 week from Course 3 Day 1 to Course 11 Day 1, and every 3 courses thereafter until disease progression beyond irRC or patient exclusion. irRC, immune-dependent response criteria in the treatment of solid tumors; MTD, maximum tolerated dose; NSCLC, non-small cell lung cancer; RP2D, recommended dose in phase II; TNBC, triple negative breast cancer.

Figure 20 shows the percentage change in target lesions over time for each of the radiographically assessed patients treated with an anti-PD-1 antibody molecule in Phase I/II studies. Patients were treated according to the following regimens: 1 mg/kg every other week, 3 mg/kg every other week, 10 mg/kg every other week, 3 mg/kg every four weeks, or 5 mg/kg every four weeks. weeks (exemplary lines in the figure are indicated by application patterns).

Figures 21A-21C show tumor scores and immunohistochemical detection of CD8+ T lymphocytes in a patient with a metastatic atypical lung carcinoid tumor with a clinical response to an anti-PD-1 antibody molecule in Phase I/II studies.

Figure 21A shows CT scan images showing patient response. The left panel shows liver metastasis before antibody treatment. The middle panel shows pseudoprogression in the liver (accompanied by a significant decrease in lesions in the lung, not shown) at the first re-staging. The right panel shows the response in all lesions at the second re-staging.

Figure 21B shows the reduction in metastatic atypical lung carcinoid tumor burden (% change from baseline) and individual foci (lesion size (nm)) in a patient.

Figure 21C shows immunohistochemical staining images showing high levels of CD8+ T lymphocytes in a tumor sample obtained from a patient during Course 2, Day 1. BL, baseline; C2D1, Course 2, Day 1.

Figure 22A shows the percentage of surviving mice after implantation of MC38 cells and administration of anti-PD-1 antibody, Compound A15 (once a week or daily) or a combination of anti-PD-1 antibody and Compound A15 (once a week or daily).

Figure 22B shows mean tumor volume in mice after implantation of MC38 cells and administration of anti-PD-1 antibody, Compound A15 (daily) or a combination of anti-PD-1 antibody and Compound A15 (daily).

Figure 22C shows mean tumor volume in mice after implantation of MC38 cells and administration of anti-PD-1 antibody, Compound A15 (once per week) or combination of anti-PD-1 antibody and Compound A15 (once per week).

Figure 23 shows the improvement over time to end point observed in animals treated with an anti-PD-L1 antibody molecule in combination with an anti-PD-1 antibody.

BRIEF DESCRIPTION OF THE TABLES

Table 1 is a table of amino acid and nucleotide sequences of mouse, chimeric and humanized anti-PD-1 antibody molecules. Antibody molecules include mouse mAb BAP049, chimeric mAbs BAP049-chi and BAP049-chi-Y, and humanized mAbs BAP049-hum01 to BAP049-hum16 and BAP049-Clone-A to BAP049-Clone-E. This table shows the amino acid and nucleotide sequences of the heavy and light chain CDR regions, 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.

Table 2 shows the amino acid and nucleotide sequences of the heavy and light chain framework regions of the humanized mAbs BAP049-hum01 to BAP049-hum16 and BAP049-Clone-A to BAP049-Clone-E.

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

Table 4 shows the amino acid sequences of the heavy and light chain leader sequences of the humanized BAP049-Clone-A to BAP049-Clone-E mAbs.

Table 5 shows exemplary PK parameters based on constant dose schedules.

Table 6 provides an exemplary list of therapeutic agents from Antigen Presenting Combinations (Category A), Effector Cell Combinations (Category B), and Anti-Tumor Immunosuppression Combinations (Category C).

Table 7 is a table of selected therapeutic agents that can be administered in combination with anti-PD-1 antibody molecules and other immunomodulators (e.g., one or more of the following: a costimulatory molecule activator and/or an immune checkpoint molecule inhibitor) described herein. Table 7 presents the following, from left to right: Compound designation of the second therapeutic agent, Structure of the compound, and Patent publication(s) in which the Compound is disclosed.

Table 8 is a table of exemplary biomarkers and a set of samples, for example, to evaluate the effectiveness of the treatments (eg, combination therapies) described in this application.

Table 9 - table of goals and expected indicators in the I/II phase of research.

Table 10 is a table of demographic data and characteristics of patients in phase I/II studies.

Table 11 shows the distribution of patients in Phase I/II studies.

Table 12 shows adverse events regardless of relationship to study drug in Phase I/II studies (any gradient observed in ≥20% of patients is a safety sample).

Table 13 shows the best overall response (based on the investigator's assessment of disease status using the RECIST v1.1 test).

DETAILED DESCRIPTION

Disclosed herein are, at least in part, antibody molecules (eg, humanized antibody molecules) that bind to programmed death 1 (PD-1) protein with high affinity and specificity. Also provided are nucleic acid molecules encoding antibody molecules, expression vectors, host cells, and methods for producing antibody molecules. Pharmaceutical compositions and dosage forms comprising antibody molecules are also provided. The anti-PD-1 antibody molecules disclosed herein may be used (alone or in combination with other agents or therapies) for the treatment, prevention and/or diagnosis of disorders such as cancers (e.g. solid tumors and soft tissue tumors) , as well as infectious diseases (for example, chronic infectious diseases or sepsis). Thus, the present application discloses compositions and methods for detecting PD-1, as well as methods for treating various disorders, including cancer and/or infectious diseases, using anti-PD-1 antibody molecules. In some embodiments, the anti-PD-1 antibody molecule is administered or administered at a constant or fixed dose.

Also disclosed herein are methods and compositions comprising a combination of two, three, or more therapeutic agents selected from one, two, or all of the following categories (i)-(iii): (i) an agent that increases antigen presentation (e.g., tumor presentation). antigen) (eg, by increasing one or more of dendritic cell activity or maturation, antigen uptake, or antigen processing); (ii) an agent that enhances an effector cell response (eg, an immune effector cell response, eg, activation and/or mobilization of B cells and/or T cells, eg, in a lymph node); or (iii) an agent that reduces tumor immunosuppression (eg, by increasing T cell infiltration and killing tumor cells). In some embodiments, the combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein). Without being limited by theory, it is believed that therapeutic methods that enhance antitumor immunity are more effective when the immune response is optimized through different targets at different stages of the immune response. Each of these steps is shown in schematic form in Figure 21. For example, methods that result in dendritic cell activation, combined with methods that enhance cellular and humoral immunity, may result in a more effective and/or prolonged therapeutic response.

Additional terms are defined below and throughout the text of the application.

When used in this application, the singular form (the articles "a" and "an" in the original text of the description) refer to one or more than one (eg, at least one) grammatical object of the article.

The term "or" is used in this application to mean the term "and/or" and is used interchangeably with it, unless the context clearly indicates otherwise.

"Approximately" and "about" usually mean the acceptable degree of error for the quantity being measured, taking into account the nature or accuracy of the measurements. The approximate degree of error is within 20 percent (%), typically within 10%, and more typically within 5% of a given value or range of values.

By "combination" or "in combination with" it is not meant that the therapy or therapeutic agents must be administered at the same time and/or included for delivery in the same dosage form, although such delivery methods are included within the scope of the present disclosure. Therapeutic agents in combination can be used simultaneously, before or after one or more other adjunctive therapies or therapeutic agents. Therapeutic agents or therapeutic protocol may be administered in any order. Typically, each agent is administered at a dose and/or schedule determined for that agent. It should also be understood that the additional therapeutic agent used in this combination may be administered together in the same composition or administered separately in different compositions. In general, it is expected that additional therapeutic agents used in combination will be used at levels that do not exceed the levels at which they are used separately. In some embodiments, the levels used in combination will be lower than those used alone.

In embodiments, the additional therapeutic agent is administered at a therapeutic dose or lower than the therapeutic dose. In some embodiments, the concentration of the second therapeutic agent that is required for inhibition, such as growth inhibition, is lower when the second therapeutic agent is used in combination with the first therapeutic agent, such as an anti-PD-1 antibody molecule, than when the second therapeutic agent is administered separately. In some embodiments, the concentration of the first therapeutic agent required for inhibition, such as growth inhibition, is lower when the first therapeutic agent is administered in combination with the second therapeutic agent than when the first therapeutic agent is administered alone. In some embodiments, in combination therapy, the concentration of the second therapeutic agent that is required for inhibition, such as growth inhibition, is lower than the therapeutic dose of the second therapeutic agent in monotherapy, for example, lower by 10-20%, 20-30%, 30- 40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90%. In some embodiments, in combination therapy, the concentration of the first therapeutic agent that is required for inhibition, such as growth inhibition, is lower than the therapeutic dose of the first therapeutic agent in monotherapy, for example, lower by 10-20%, 20-30%, 30- 40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90%.

The term "inhibition", "inhibitor", or "antagonist" includes a reduction in some parameter, such as activity, of a given molecule, such as an immune checkpoint inhibitor. For example, the term includes inhibition of an activity, such as PD-1 or PD-L1 activity, by at least 5%, 10%, 20%, 30%, 40%, or more. Thus inhibition should not be 100%.

The term "activation", "activator", or "agonist" includes an increase in some parameter, such as activity, of a given molecule, such as a costimulatory molecule. For example, this term includes an increase in activity, such as costimulatory activity, at least 5%, 10%, 25%, 50%, 75% or more.

The term "anti-cancer effect" refers to a biological effect that can manifest itself in a variety of ways, including, but not limited to, e.g., reduction in tumor volume, reduction in the number of cancer cells, reduction in the number of metastases, increase in lifespan, reduction in cancer cell proliferation, reduction in cancer survival. cells or a reduction in the severity of various physiological symptoms associated with cancer. "Anti-cancer activity" can also be manifested in the ability of peptides, polynucleotides, cells and antibodies to prevent the primary occurrence of cancer.

The term "anti-tumor effect" refers to a biological effect that can be manifested in a variety of ways, including, but not limited to, for example, reduction in tumor volume, reduction in the number of tumor cells, reduction in tumor cell proliferation, or reduction in tumor cell survival.

The term "cancer" refers to a disease characterized by the rapid and uncontrolled growth of aberrant cells. Cancer cells can spread locally or, through the bloodstream and lymphatic system, to other parts of the body. Examples of various forms of cancer are described herein and include, but are not limited to, breast cancer, prostate cancer, ovarian cancer, cervical cancer, skin cancer, pancreatic cancer, colon and rectal cancer, kidney cancer, liver cancer, cancer brain, lymphoma, leukemia, lung cancer, etc. The terms "tumor" and "cancer" are used interchangeably in this application, for example, both terms cover solid and hemoblastic, for example, diffuse or circulating tumors. When used in this application, the term "cancer" or "tumor" includes precancerous tumors, as well as cancer and malignant tumors.

The term "antigen presenting cell" or "APC" refers to a cell of the immune system, such as a helper cell (e.g., B cell, dendritic cell, etc.) that presents a foreign antigen bound to major histocompatibility complex (MHC) molecules, on its surface. T cells can recognize such complexes using their T cell receptors (TCRs). APCs process antigens and present them to T cells.

The term "costimulatory molecule" refers to a cognate binding partner on a T cell that specifically binds to a costimulatory ligand, thereby mediating a costimulatory response of the T cell, such as, but not limited to, proliferation. Costimulatory molecules are cell surface molecules other than antigen receptors or their ligands, which are required for an effective immune response. Costimulatory molecules include, but are not limited to, MHC class I molecule, TNF receptor proteins, immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocyte-activating molecules (SLAM proteins), activating NK cell receptors, BTLA, Toll Ligand receptor, OX40 , CD2, CD7, CD27, CD28, CD30, CD40, CD, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CD, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8 alpha, CD8 beta, IL2R beta, IL2R gamma, IL7R alpha, 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), LTB R, LAT, GADS, SLP-76, PAG/Cbp, CD19a, and a ligand that specifically binds to CD83.

"Immune effector cell" or "effector cell", as the term is used in this application, refers to a cell that is involved in an immune response, for example, in the stimulation of 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 cells (NKT), mast cells, and myeloid phagocytes. origin.

An "immune effector" or "effector" "function" or "response", as the term is used in this application, refers to a function or response, for example, of an immune effector cell that enhances or stimulates an immune attack on a target cell. For example, an immune effector function or response refers to a property of a T or NK cell that causes the killing or inhibition of growth or proliferation of a target cell. In the case of a T cell, primary stimulation and costimulation are examples of an immune effector function or response.

The term "effector function" refers to a specialized function of a cell. The effector function of the T cell, for example, may be cytolytic activity or helper activity involving the secretion of cytokines.

As used herein, the terms "treat" and "treatment" refer to the reduction or amelioration of the progression, severity and/or duration of a disorder, e.g., a proliferative disorder, or to the improvement of one or more symptoms (preferably one or more overt symptoms) of a disorder, caused by the use of one or more therapies. In some embodiments, the terms "treat" and "treatment" refer to attenuation of at least one measurable physical parameter of a proliferative disorder, such as tumor growth, that is not necessarily apparent to the patient. In other embodiments, the terms "treat" and "treatment" refer to inhibiting the progression of a proliferative disorder, either physical, such as by stabilizing an overt symptom, or physiological, such as by stabilizing a physical parameter, or both. In other embodiments, the terms "treat" and "treatment" refer to reducing or stabilizing the size of a tumor or the number of cancer cells.

Compositions and methods of the present invention encompass polypeptides and nucleic acids having certain sequences, or sequences substantially identical or similar to them, for example, sequences at least 85%, 90%, 95% or more identical to a certain sequence. In the case of an amino acid sequence, the term "substantially identical" is used herein to refer to the first amino acid that contains a sufficient or minimum number of amino acid residues that are: i) identical or ii) are conservative substitutions of aligned amino acid residues in the second amino acid sequence, resulting in the first and second amino acid sequences may share a common structural domain and/or a common functional activity. For example, amino acid sequences that contain a common structural domain having at least about 85%, 90%. 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with a reference sequence, for example, the sequence presented in this application.

In the case of a nucleotide sequence, 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 aligned nucleotides in a second nucleic acid sequence such that the first and second nucleotide sequences encode a polypeptide, having a common functional activity, or encode a common structural domain of the polypeptide or a common functional activity of the polypeptide. For example, nucleotide sequences that have at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with a reference sequence, such as the sequence provided in this application.

The term "functional variant" refers to polypeptides that have an amino acid sequence substantially identical to the natural sequence or encoded by a substantially identical nucleotide sequence, and may have one or more activities of the natural sequence.

Calculations of homology or sequence identity (the terms are used interchangeably in this application) are performed as follows.

To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison (e.g., gaps can be introduced in one or both of the first and second amino acid sequences or nucleic acid sequences for optimal alignment, while non-homologous sequences can not be 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% and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. The amino acid residues or nucleotides at the corresponding amino acid positions or nucleotide positions are then 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, then the molecules at that position are identical (as used herein, “identity” of an amino acid or nucleic acid is equivalent to “homology” of an amino acid or nucleic acid). acids).

The percent identity between two sequences depends on the number of identical positions in those sequences, taking into account the number of gaps and the length of each gap that must be entered to optimally align the two sequences.

Sequence comparison and determination of percent identity between two sequences can be performed using a mathematical algorithm. In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch (1970) J. Mol. Biol. 48:444-453 algorithm, which is part of the GAP program in the GCG software package (available at www.gcg.com), when using a Blossum 62 Matrix or a PAM250 Matrix, and a Gap Penalty of 16, 14, 12, 10, 8, 6, or 4, and a Gap Continue Penalty of 1, 2, 3, 4, 5 or 6. In another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at www.gcg.com), using the NWSgapdna.CMP matrix, a gap penalty of 40, 50, 60 , 70, or 80, and a gap continuation penalty of 1, 2, 3, 4, 5, or 6. An especially preferred set of parameters (and the set to be used unless otherwise noted) is the Blossum 62 substitution matrix with a penalty for the introduction of a gap 12, a penalty for extending a gap 4 and a penalty for a gap with a frame shift 5.

The percentage identity between two amino acid or nucleotide sequences can be determined using the algorithm of Y. Meyers and W. Miller (E. Meyers and W. Miller (1989) CABIOS, 4: 1117), which is included in the ALIGN program (version 2.0), with using a table of weights for substitutions of residues RAM120, gap extension penalty 12 and gap insertion penalty 4.

The nucleic acid and protein sequences described herein can be used as a "query sequence" to perform searches in public databases, for example, to identify other family members or related sequences. Such searches can be performed using the NBLAST program and XBLAST (version 2.0) according to Altschul et al. (1990), J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed using the NBLAST program, score=100, wordlength=12, to obtain nucleotide sequences homologous to nucleic acid molecules (SEQ ID NO: 1) according to the invention. A BLAST search for proteins can be performed using the XBLAST program, score=50, wordlength=3, to obtain amino acid sequences homologous to the protein molecules of the invention. To obtain gapped alignments for comparison purposes, the Gapped BLAST program described in Altschul et al., (1997) Nucleic Acids Res. 25:3389-3402. When using the BLAST and Gapped BLAST programs, you can use the default settings of the respective programs (for example, XBLAST and NBLAST). See www.ncbi.nlm.nih.gov.

As used herein, the term "hybridizes under low stringency, medium stringency, high stringency, or very high stringency conditions" describes hybridization and washing conditions. Guidelines for conducting 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. This handbook describes aqueous and non-aqueous methods, and any of them can be used. The specific hybridization conditions referred to herein are as follows: 1) low stringency hybridization conditions in 6× sodium chloride/sodium citrate (SSC) at approximately 45° C., followed by two subsequent washes in 0.2× SSC, 0.1% SDS at least 50°C (in low stringency conditions, the temperature during washings can be increased up to 55°C); 2) medium stringency hybridization conditions in 6x SSC at approximately 45°C, followed by one or more washes in 0.2x SSC, 0.1% SDS at 60°C; 3) high stringency hybridization conditions in 6x SSC at approximately 45°C, followed by one or more washes in 0.2x SSC, 0.1% SDS at 65°C; and preferably 4) very stringent hybridization conditions: 0.5M sodium phosphate, 7% SDS at 65°C, followed by one or more washes in 0.2x SSC, 1% SDS at 65°C. The very stringent conditions (4) are the preferred conditions and the conditions to be used unless otherwise noted.

It should be understood that the molecules of the present invention may have additional conservative or non-essential amino acid substitutions that do not significantly affect their function.

The term "amino acid" is intended to encompass all molecules, both natural and synthetic, that simultaneously contain an amino functional group and an acid functional group, and may be incorporated into a polymer of naturally occurring amino acids. Examples of amino acids include natural amino acids, their analogues, derivatives and related compounds; amino acid analogs having modified side chains; and all their stereoisomers of any of the previous. When used in the invention, the term "amino acid" includes D - and L - optical isomers and peptidomimetics.

A "conservative amino acid substitution" is one 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 have been defined in the prior art. Such families include amino acids with basic side chains (eg, lysine, arginine, histidine), acidic side chains (eg, aspartic acid, glutamic acid), uncharged polar side chains (eg, glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (eg, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (eg, threonine, valine, isoleucine), and aromatic side chains (eg, tyrosine, phenylalanine , tryptophan, histidine).

The terms "polypeptide", "peptide" and "protein" (in the case of a single chain) are used interchangeably in this application to refer to amino acid polymers of any length. Such a polymer may be linear or branched, it may include modified amino acids, and it may be interrupted by non-amino acids. The terms also cover an amino acid polymer that has been modified; for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation such as conjugation with a labeling moiety. The polypeptide may be isolated from natural sources, may be obtained by recombinant methods from a eukaryotic or prokaryotic host, or may be the product of synthetic procedures.

The terms "nucleic acid", "nucleic acid sequence", "nucleotide sequence" or "polynucleotide sequence" and "polynucleotide" are used interchangeably. These terms refer to the polymeric form of nucleotides of any length, deoxyribonucleotides or ribonucleotides, or analogs thereof. The polynucleotide may be single stranded or double stranded, and if it is single stranded, it may be a coding strand or a non-coding (antisense) strand. The polynucleotide may contain modified nucleotides such as methylated nucleotides and nucleotide analogs. The sequence of nucleotides may be interrupted by non-nucleotide components. The polynucleotide may be further modified after polymerization, for example by conjugation with a labeling moiety. The nucleic acid may be a recombinant polynucleotide or a polynucleotide of genomic, cDNA, semi-synthetic or synthetic origin, which is either not found in nature or linked to another polynucleotide in a non-natural configuration.

The term "isolated" as used herein refers to material that has been removed from its original or native environment (eg, from the natural environment, if such material is natural). For example, a natural polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide separated by human intervention from some or all of the accompanying materials in the natural system is isolated. Such polynucleotides may be part of a vector, and/or such polynucleotides or polypeptides may be part of a composition and yet be isolated in such a vector or composition because they will not be part of the environment in which they occur naturally.

Various aspects of the present invention are described in more detail below. Additional definitions are given in the description text.

Exemplary Combinations of Antigen-Presenting Combinations, Effector Cell Combinations, and Anti-Tumor Immunosuppression Combinations

This application provides exemplary combinations of therapeutic agents from two or more of the antigen-presenting category (A), the effector cell category (B), and the anti-tumor immunosuppression category (C).

Table 6: List of therapeutic agents in categories (A)-(C)

A= Antigen presentation B= Effector cell C= Against tumor immunosuppression 1 STING agonist GITR agonist PD-1 inhibitor 2 TLR agonist PD-1 inhibitor PD-L1 inhibitor 3 TIM-3 modulator PD-L1 inhibitor LAG-3 inhibitor four VEGFR inhibitor IAP inhibitor TIM-3 inhibitor five c-MET inhibitor EGFR inhibitor GITR inhibitor 6 TGFb inhibitor mTOR inhibitor CSF-1/1R inhibitor 7 IDO/TDO inhibitor IL-15 agonist IL-17 inhibitor 8 A2AR antagonist CTLA-4 inhibitor IL-1β inhibitor nine Oncolytic viruses Bispecific T cell activators CXCR2 inhibitor 10 Scaffold vaccines CD40 agonist PI3K inhibitor-γ, -δ eleven Bispecific T cell activators OX40 agonist BAFF-R inhibitor 12 CD27 agonist MALT-1/BTK inhibitor 13 JAK inhibitor fourteen CRTH2 inhibitor 15 VEGFR inhibitor sixteen IL-15 agonist 17 TGFb inhibitor antibody eighteen IDO/TDO inhibitor nineteen A2AR antagonist 20 CTLA-4 inhibitor 21 PFKFB3 inhibitor

In some embodiments, combinations of the present invention include one or more of the following:

A1B1, A1B2, A1B3, A1B4, A1B5, A1B6, A1B7, A1B8, A1B9, A1B10, A1B11, A1B12, A2B1, A2B2, A2B3, A2B4, A2B5, A2B6, A2B7, A2B8, A2B9, A2B10, A2B11, A3B1, A3B2, A3B3, A3B4, A3B5, A3B6, A3B7, A3B8, A3B9, A3B10, A3B11, A3B12, A4B1, A4B2, A4B3, A4B4, A4B5, A4B6, A4B7, A4B8, A4B9, A4B10, A4B11, A4B12, A5B2,1, A5B3, A5B4, A5B5, A5B6, A5B7, A5B8, A5B9, A5B10, A5B11, A5B12, A6B1, A6B2, A6B3, A6B4, A6B5, A6B6, A6B7, A6B8, A6B9, A6B10, A6B11, A6B12, A7B1, A7B3, A7B4, A7B5, A7B6, A7B7, A7B8, A7B9, A7B10, A7B11, A7B12, A8B1, A8B2, A8B3, A8B4, A8B5, A8B6, A8B7, A8B8, A8B9, A8B10, A8B11, A8B12, A9B1, A9B2, A9B4, A9B4, A9B5 A9B6 A9B7 A9B8 A9B9 A9B10 A9B11 A9B12 A10B1 A10B2 A10B3 A10B4 A10B5 A10B6 A10B7 A10B8 A10B9 A10B10 A10B11 A10B12 A11B3 A11B6, A11B7, A11B8, A11B9, A11B10, A11B11, A11B12, A1C1, A1C2, A1C3, A1C4, A1C5, A1C6, A1C7, A1C8, A1C9, A1C10, A1C11, A1C12, A1C13, A1C14, A1C16, A1C15, A1C19, A1C20, A1C21, A2C1, A2C2, A 2C3 A2C4 A2C5 A2C6 A2C7 A2C8 A2C9 A2C10 A2C11 A2C12 A2C13 A2C14 A2C15 A2C16 A2C17 A2C18 A2C19 A2C20 A2C21 A3C1 A3C2 A3C3 A3C4 A3C7 A3C8 A3C9 A3C10 A3C11 A3C12 A3C13 A3C14 A3C15 A3C16 A3C17 A3C18 A3C19 A3C20 A3C21 A4C1 A4C2 A4C3 A4C4 A4C5 A4C6 A4C0 A4C8 A4C11, A4C12, A4C13, A4C14, A4C15, A4C16, A4C17, A4C18, A4C19, A4C20, A4C21, A5C1, A5C2, A5C3, A5C4, A5C5, A5C6, A5C7, A5C8, A5C9, A5C10, A5C13, A5C12, A5C1 A5C15, A5C16, A5C17, A5C18, A5C19, A5C20, A5C21, A6C1, A6C2, A6C3, A6C4, A6C5, A6C6, A6C7, A6C8, A6C9, A6C10, A6C11, A6C12, A6C13, A6C14, A6C16, A6C18 A6C19, A6C20, A6C21, A7C1, A7C2, A7C3, A7C4, A7C5, A7C6, A7C7, A7C8, A7C9, A7C10, A7C11, A7C12, A7C13, A7C14, A7C15, A7C16, A7C17, A7C18, A7C19, A7C21 A8C2, A8C3, A8C4, A8C5, A8C6, A8C7, A8C8, A8C9, A8C10, A8C11, A8C12, A8C13, A8C14, A8C15, A8C16, A8C17, A8C18, A8C19, A8C20, A8C21, A9C1, A9C2, A9C3 A9C6, A9C7, A9C8, A9C9, A9C10, A9C11, A9C12, A9C13, A9C14, A9C15, A9C16, A9C17, A9C18, A9C19, A9C20, A9C21, A10C1, A10C2, A10C3, A10C4, A10C5, A10C6, A10C7, A10C8, A10C9, A10C10, A10C11, A10C12, A10C13, A10C14, A10C15, A10C16, A10C17, A10C18, A10C19, A10C20, A10C21, A11C1, A11C2, A11C3, A11C4, A11C5, A11C6, A11C7, A11C8, A11C9, A11C10, A11C11, A11C12, A11C13, A11C14, A11C15, A11C16, A11C17, A11C18, A11C19, A11C20, A11C21, B1C1, B1C2, B1C3, B1C4, B1C5, B1C6, B1C7, B1C8, B1C9, B1C10, B1C11, B1C12, B1C13, B1C14, B1C15, B1C16, B1C18 B1C21, B2C1, B2C2, B2C3, B2C4, B2C5, B2C6, B2C7, B2C8, B2C9, B2C10, B2C11, B2C12, B2C13, B2C14, B2C15, B2C16, B2C17, B2C18, B2C19, B2C20, B2C21, B2C B3C4 B3C5 B3C6 B3C7 B3C8 B3C9 B3C10 B3C11 B3C12 B3C13 B3C14 B3C15 B3C16 B3C17 B3C18 B3C19 B3C20 B3C21 B4C1 B4C2 B4C3 B4C4 B4C5 B4C8, B4C9, B4C10, B4C11, B4C12, B4C13, B4C14, B4C15, B4C16, B4C17, B4C18, B4C19, B4C20, B4C21, B5C1, B5C2, B5C3, B5C4, B5C5, B5C6, B5C 7, B5C8, B5C9, B5C10, B5C11, B5C12, B5C13, B5C14, B5C15, B5C16, B5C17, B5C18, B5C19, B5C20, B5C21, B6C1, B6C2, B6C3, B6C4, B6C5, B6C6, B6C7, B6, B6C8 B6C11, B6C12, B6C13, B6C14, B6C15, B6C16, B6C17, B6C18, B6C19, B6C20, B6C21, B7C1, B7C2, B7C3, B7C4, B7C5, B7C6, B7C7, B7C8, B7C9, B7C10, B7C13, B7C12 B7C15 B7C16 B7C17 B7C18 B7C19 B7C20 B7C21 B8C1 B8C2 B8C3 B8C4 B8C5 B8C6 B8C7 B8C8 B8C9 B8C10 B8C11 B8C12 B8C13 B8C14 B8C16 B8C15 B8C19, B8C20, B8C21, B9C1, B9C2, B9C3, B9C4, B9C5, B9C6, B9C7, B9C8, B9C9, B9C10, B9C11, B9C12, B9C13, B9C14, B9C15, B9C16, B9C17, B9C18, B9C10, B9C21 B10C2, B10C3, B10C4, B10C5, B10C6, B10C7, B10C8, B10C9, B10C10, B10C11, B10C12, B10C13, B10C14, B10C15, B10C16, B10C17, B10C18, B10C19, B10C20, B10C21, B11C1, B11C2, B11C3, B11C4, B11C5, B11C6 B11C7 B11C8 B11C9 B11C10 B11C11 B11C12 B11C13 B11C14 B11C15 B11C16 B11C17 B11C18 B11C19 B11C20 B11C21 B12C1 B12C2 B12C3 B1 C5, B12C6, B12C7, B12C8, B12C9, B12C10, B12C11, B12C12, B12C13, B12C14, B12C15, B12C16, B12C17, B12C18, B12C19, B12C20, B12C21, A1B1C1, A1B1C2, A1B1C3, A1B1C4, A1B1C5, A1B1C6, A1B1C7, A1B1C8, A1B1C9, A1B1C10, A1B1C11, A1B1C12, A1B1C13, A1B1C14, A1B1C15, A1B1C16, A1B1C17, A1B1C18, A1B1C19, A1B1C20, A1B1C21, A1B2C1, A1B2C2, A1B2C3, A1B2C4, A1B2C5, A1B2C6, A1B2C7, A1B2C8, A1B2C9, A1B2C10, A1B2C11, A1B2C12, A1B2C13, A1B2C14, A1B2C15, A1B2C16, A1B2C17, A1B2C18, A1B2C19, A1B2C20, A1B2C21, A1B3C1, A1B3C2, A1B3C3, A1B3C4, A1B3C5, A1B3C6, A1B3C7, A1B3C8, A1B3C9, A1B3C10, A1B3C11, A1B3C12, A1B3C13, A1B3C14, A1B3C15, A1B3C16, A1B3C17, A1B3C18, A1B3C19, A1B3C20, A1B3C21, A1B4C1, A1B4C2, A1B4C3, A1B4C4, A1B4C5, A1B4C6, A1B4C7, A1B4C8, A1B4C9, A1B4C10, A1B4C11, A1B4C12, A1B4C13, A1B4C14, A1B4C15, A1B4C16, A1B4C17, A1B4C18, A1B4C19, A1B4C20, A1B4C21, A1B5C1, A1B5C2, A1B5C3, A1B5C4, A1B5C5, A1B5C6, A1B5C7, A1B5C8, A1B5C9, A1B5C10, A1B5C11, A1B5C12, A1B5C13, A1B5C14, A1B5C15, A1B5B16 A1B5C19, A1B5C20, A1B5C21, A1B6C1, A1B6C2, A1B6C3, A1B6C4, A1B6C5, A1B6C6, A1B6C7, A1B6C8, A1B6C9, A1B6C10, A1B6C11, A1B6C12, A1B6C13, A1B6C14, A1B6C15, A1B6C16, A1B6C17, A1B6C18, A1B6C19, A1B6C20, A1B6C21, A1B7C1, A1B7C2, A1B7C3, A1B7C4, A1B7C5, A1B7C6, A1B7C7, A1B7C8, A1B7C9, A1B7C10, A1B7C11, A1B7C12, A1B7C13, A1B7C14, A1B7C15, A1B7C16, A1B7C17, A1B7C18, A1B7C19, A1B7C20, A1B7C21, A1B8C1, A1B8C2, A1B8C3, A1B8C4, A1B8C5, A1B8C6, A1B8C7, A1B8C8, A1B8C9, A1B8C10, A1B8C11, A1B8C12, A1B8C13, A1B8C14, A1B8C15, A1B8C16, A1B8C17, A1B8C18, A1B8C19, A1B8C20, A1B8C21, A1B9C1, A1B9C2, A1B9C3, A1B9C4, A1B9C5, A1B9C6, A1B9C7, A1B9C8, A1B9C9, A1B9C10, A1B9C11, A1B9C12, A1B9C13, A1B9C14, A1B9C15, A1B9C16, A1B9C17, A1B9C18, A1B9C19, A1B9C20, A1B9C21, A1B10C1, A1B10C2, A1B10C3, A1B10C4, A1B10C5, A1B10C6, A1B10C7, A1B10C8, A1B10C9, A1B10C10, A1B10C11, A1B10C12, A1B10C13, A1B10C14, A1B10C15, A1B10C16, A1B10C17, A1B10C18, A1B10C19, A1B10C20, A1B10C21, A1B11C1, A1B11C2, A1B11C3, A1B11C4, A1B11C5, A1B11C 6, A1B11C7, A1B11C8, A1B11C9, A1B11C10, A1B11C11, A1B11C12, A1B11C13, A1B11C14, A1B11C15, A1B11C16, A1B11C17, A1B11C18, A1B11C19, A1B11C20, A1B11C21, A1B12C1, A1B12C2, A1B12C3, A1B12C4, A1B12C5, A1B12C6, A1B12C7, A1B12C8, A1B12C9, A1B12C10, A1B12C11, A1B12C12, A1B12C13, A1B12C14, A1B12C15, A1B12C16, A1B12C17, A1B12C18, A1B12C19, A1B12C20, A1B12C21, A2B1C1, A2B1C2, A2B1C3, A2B1C4, A2B1C5, A2B1C6, A2B1C7, A2B1C8, A2B1C9, A2B1C10, A2B1C11, A2B1C12, A2B1C13, A2B1C14, A2B1C15, A2B1C16, A2B1C17, A2B1C18, A2B1C19, A2B1C20, A2B1C21, A2B2C1, A2B2C2, A2B2C3, A2B2C4, A2B2C5, A2B2C6, A2B2C7, A2B2C8, A2B2C9, A2B2C10, A2B2C11, A2B2C12, A2B2C13, A2B2C14, A2B2C15, A2B2C16, A2B2C17, A2B2C18, A2B2C19, A2B2C20, A2B2C21, A2B3C1, A2B3C2, A2B3C3, A2B3C4, A2B3C5, A2B3C6, A2B3C7, A2B3C8, A2B3C9, A2B3C10, A2B3C11, A2B3C12, A2B3C13, A2B3C14, A2B3C15, A2B3C16, A2B3C17, A2B3C18, A2B3C19, A2B3C20, A2B3C21, A2B4C1, A2B4C2, A2B4C3, A2B4C4, A2B4C5, A2B4C6, A2B4C7, A2B4C8, A2B4C9, A2B4C10, A2B4C11, A2B4C12, A2B4C13, A 2B4C14, A2B4C15, A2B4C16, A2B4C17, A2B4C18, A2B4C19, A2B4C20, A2B4C21, A2B5C1, A2B5C2, A2B5C3, A2B5C4, A2B5C5, A2B5C6, A2B5C7, A2B5C8, A2B5C9, A2B5C10, A2B5C11, A2B5C12, A2B5C13, A2B5C14, A2B5C15, A2B5C16, A2B5C17, A2B5C18, A2B5C19, A2B5C20, A2B5C21, A2B6C1, A2B6C2, A2B6C3, A2B6C4, A2B6C5, A2B6C6, A2B6C7, A2B6C8, A2B6C9, A2B6C10, A2B6C11, A2B6C12, A2B6C13, A2B6C14, A2B6C15, A2B6C16, A2B6C17, A2B6C18, A2B6C19, A2B6C20, A2B6C21, A2B7C1, A2B7C2, A2B7C3, A2B7C4, A2B7C5, A2B7C6, A2B7C7, A2B7C8, A2B7C9, A2B7C10, A2B7C11, A2B7C12, A2B7C13, A2B7C14, A2B7C15, A2B7C16, A2B7C17, A2B7C18, A2B7C19, A2B7C20, A2B7C21, A2B8C1, A2B8C2, A2B8C3, A2B8C4, A2B8C5, A2B8C6, A2B8C7, A2B8C8, A2B8C9, A2B8C10, A2B8C11, A2B8C12, A2B8C13, A2B8C14, A2B8C15, A2B8C16, A2B8C17, A2B8C18, A2B8C19, A2B8C20, A2B8C21, A2B9C1, A2B9C2, A2B9C3, A2B9C4, A2B9C5, A2B9C6, A2B9C7, A2B9C8, A2B9C9, A2B9C10, A2B9C11, A2B9C12, A2B9C13, A2B9C14, A2B9C15, A2B9C16, A2B9C17, A2B9C18, A2B9C19, A2B9C20, A2B9C21, A2B10C1, A2B10C2, A2B10C3 B10C4, A2B10C5, A2B10C6, A2B10C7, A2B10C8, A2B10C9, A2B10C10, A2B10C11, A2B10C12, A2B10C13, A2B10C14, A2B10C15, A2B10C16, A2B10C17, A2B10C18, A2B10C19, A2B10C20, A2B10C21, A2B11C1, A2B11C2, A2B11C3, A2B11C4, A2B11C5, A2B11C6, A2B11C7, A2B11C8, A2B11C9, A2B11C10, A2B11C11, A2B11C12, A2B11C13, A2B11C14, A2B11C15, A2B11C16, A2B11C17, A2B11C18, A2B11C19, A2B11C20, A2B11C21, A2B12C1, A2B12C2, A2B12C3, A2B12C4, A2B12C5, A2B12C6, A2B12C7, A2B12C8, A2B12C9, A2B12C10, A2B12C11, A2B12C12, A2B12C13, A2B12C14, A2B12C15, A2B12C16, A2B12C17, A2B12C18, A2B12C19, A2B12C20, A2B12C21, A3B1C1, A3B1C2, A3B1C3, A3B1C4, A3B1C5, A3B1C6, A3B1C7, A3B1C8, A3B1C9, A3B1C10, A3B1C11, A3B1C12, A3B1C13, A3B1C14, A3B1C15, A3B1C16, A3B1C17, A3B1C18, A3B1C19, A3B1C20, A3B1C21, A3B2C1, A3B2C2, A3B2C3, A3B2C4, A3B2C5, A3B2C6, A3B2C7, A3B2C8, A3B2C9, A3B2C10, A3B2C11, A3B2C12, A3B2C13, A3B2C14, A3B2C15, A3B2C16, A3B2C17, A3B2C18, A3B2C19, A3B2C20, A3B2C21, A3B3C1, A3B3C2, A3B3C3, A3B3C4, A3B3C5, A3B3C6, A3B3C7, A3B3C8, A3 B3C9, A3B3C10, A3B3C11, A3B3C12, A3B3C13, A3B3C14, A3B3C15, A3B3C16, A3B3C17, A3B3C18, A3B3C19, A3B3C20, A3B3C21, A3B4C1, A3B4C2, A3B4C3, A3B4C4, A3B4C5, A3B4C6, A3B4C7, A3B4C8, A3B4C9, A3B4C10, A3B4C11, A3B4C12, A3B4C13, A3B4C14, A3B4C15, A3B4C16, A3B4C17, A3B4C18, A3B4C19, A3B4C20, A3B4C21, A3B5C1, A3B5C2, A3B5C3, A3B5C4, A3B5C5, A3B5C6, A3B5C7, A3B5C8, A3B5C9, A3B5C10, A3B5C11, A3B5C12, A3B5C13, A3B5C14, A3B5C15, A3B5C16, A3B5C17, A3B5C18, A3B5C19, A3B5C20, A3B5C21, A3B6C1, A3B6C2, A3B6C3, A3B6C4, A3B6C5, A3B6C6, A3B6C7, A3B6C8, A3B6C9, A3B6C10, A3B6C11, A3B6C12, A3B6C13, A3B6C14, A3B6C15, A3B6C16, A3B6C17, A3B6C18, A3B6C19, A3B6C20, A3B6C21, A3B7C1, A3B7C2, A3B7C3, A3B7C4, A3B7C5, A3B7C6, A3B7C7, A3B7C8, A3B7C9, A3B7C10, A3B7C11, A3B7C12, A3B7C13, A3B7C14, A3B7C15, A3B7C16, A3B7C17, A3B7C18, A3B7C19, A3B7C20, A3B7C21, A3B8C1, A3B8C2, A3B8C3, A3B8C4, A3B8C5, A3B8C6, A3B8C7, A3B8C8, A3B8C9, A3B8C10, A3B8C11, A3B8C12, A3B8C13, A3B8C14, A3B8C15, A3B8C16, A3B8C17, A3B8C18, A3B8C19, A3B8 C20, A3B8C21, A3B9C1, A3B9C2, A3B9C3, A3B9C4, A3B9C5, A3B9C6, A3B9C7, A3B9C8, A3B9C9, A3B9C10, A3B9C11, A3B9C12, A3B9C13, A3B9C14, A3B9C15, A3B9C16, A3B9C17, A3B9C18, A3B9C19, A3B9C20, A3B9C21, A3B10C1, A3B10C2, A3B10C3, A3B10C4, A3B10C5, A3B10C6, A3B10C7, A3B10C8, A3B10C9, A3B10C10, A3B10C11, A3B10C12, A3B10C13, A3B10C14, A3B10C15, A3B10C16, A3B10C17, A3B10C18, A3B10C19, A3B10C20, A3B10C21, A3B11C1, A3B11C2, A3B11C3, A3B11C4, A3B11C5, A3B11C6, A3B11C7, A3B11C8, A3B11C9, A3B11C10, A3B11C11, A3B11C12, A3B11C13, A3B11C14, A3B11C15, A3B11C16, A3B11C17, A3B11C18, A3B11C19, A3B11C20, A3B11C21, A3B12C1, A3B12C2, A3B12C3, A3B12C4, A3B12C5, A3B12C6, A3B12C7, A3B12C8, A3B12C9, A3B12C10, A3B12C11, A3B12C12, A3B12C13, A3B12C14, A3B12C15, A3B12C16, A3B12C17, A3B12C18, A3B12C19, A3B12C20, A3B12C21, A4B1C1, A4B1C2, A4B1C3, A4B1C4, A4B1C5, A4B1C6, A4B1C7, A4B1C8, A4B1C9, A4B1C10, A4B1C11, A4B1C12, A4B1C13, A4B1C14, A4B1C15, A4B1C16, A4B1C17, A4B1C18, A4B1C19, A4B1C20, A4B1C21, A4B2C1, A4B2C2, A4B2C3, A4B2C4, A4B2C5, A4B2C6, A4B2C7, A4B2C8, A4B2C9, A4B2C10, A4B2C11, A4B2C12, A4B2C13, A4B2C14, A4B2C15, A4B2C16, A4B2C17, A4B2C18, A4B2C19, A4B2C20, A4B2C21, A4B3C1, A4B3C2, A4B3C3, A4B3C4, A4B3C5, A4B3C6, A4B3C7, A4B3C8, A4B3C9, A4B3C10, A4B3C11, A4B3C12, A4B3C13, A4B3C14, A4B3C15, A4B3C16, A4B3C17, A4B3C18, A4B3C19, A4B3C20, A4B3C21, A4B4C1, A4B4C2, A4B4C3, A4B4C4, A4B4C5, A4B4C6, A4B4C7, A4B4C8, A4B4C9, A4B4C10, A4B4C11, A4B4C12, A4B4C13, A4B4C14, A4B4C15, A4B4C16, A4B4C17, A4B4C18, A4B4C19, A4B4C20, A4B4C21, A4B5C1, A4B5C2, A4B5C3, A4B5C4, A4B5C5, A4B5C6, A4B5C7, A4B5C8, A4B5C9, A4B5C10, A4B5C11, A4B5C12, A4B5C13, A4B5C14, A4B5C15, A4B5C16, A4B5C17, A4B5C18, A4B5C19, A4B5C20, A4B5C21, A4B6C1, A4B6C2, A4B6C3, A4B6C4, A4B6C5, A4B6C6, A4B6C7, A4B6C8, A4B6C9, A4B6C10, A4B6C11, A4B6C12, A4B6C13, A4B6C14, A4B6C15, A4B6C16, A4B6C17, A4B6C18, A4B6C19, A4B6C20, A4B6C21, A4B7C1, A4B7C2, A4B7C3, A4B7C4, A4B7C5, A4B7C6, A4B7C7, A4B7C8, A4B7C9, A4B7C10, A4B7C11, A4B7C12, A4B7C13, A4B7C14, A4B7C1 5, A4B7C16, A4B7C17, A4B7C18, A4B7C19, A4B7C20, A4B7C21, A4B8C1, A4B8C2, A4B8C3, A4B8C4, A4B8C5, A4B8C6, A4B8C7, A4B8C8, A4B8C9, A4B8C10, A4B8C11, A4B8C12, A4B8C13, A4B8C14, A4B8C15, A4B8C16, A4B8C17, A4B8C18, A4B8C19, A4B8C20, A4B8C21, A4B9C1, A4B9C2, A4B9C3, A4B9C4, A4B9C5, A4B9C6, A4B9C7, A4B9C8, A4B9C9, A4B9C10, A4B9C11, A4B9C12, A4B9C13, A4B9C14, A4B9C15, A4B9C16, A4B9C17, A4B9C18, A4B9C19, A4B9C20, A4B9C21, A4B10C1, A4B10C2, A4B10C3, A4B10C4, A4B10C5, A4B10C6, A4B10C7, A4B10C8, A4B10C9, A4B10C10, A4B10C11, A4B10C12, A4B10C13, A4B10C14, A4B10C15, A4B10C16, A4B10C17, A4B10C18, A4B10C19, A4B10C20, A4B10C21, A4B11C1, A4B11C2, A4B11C3, A4B11C4, A4B11C5, A4B11C6, A4B11C7, A4B11C8, A4B11C9, A4B11C10, A4B11C11, A4B11C12, A4B11C13, A4B11C14, A4B11C15, A4B11C16, A4B11C17, A4B11C18, A4B11C19, A4B11C20, A4B11C21, A4B12C1, A4B12C2, A4B12C3, A4B12C4, A4B12C5, A4B12C6, A4B12C7, A4B12C8, A4B12C9, A4B12C10, A4B12C11, A4B12C12, A4B12C13, A4B12C14, A4B12C15, A4B12C16, A4B12C17, A4B12C18, A4B12C19, A4B12C20, A4B12C21, A5B1C1, A5B1C2, A5B1C3, A5B1C4, A5B1C5, A5B1C6, A5B1C7, A5B1C8, A5B1C9, A5B1C10, A5B1C11, A5B1C12, A5B1C13, A5B1C14, A5B1C15, A5B1C16, A5B1C17, A5B1C18, A5B1C19, A5B1C20, A5B1C21, A5B2C1, A5B2C2, A5B2C3, A5B2C4, A5B2C5, A5B2C6, A5B2C7, A5B2C8, A5B2C9, A5B2C10, A5B2C11, A5B2C12, A5B2C13, A5B2C14, A5B2C15, A5B2C16, A5B2C17, A5B2C18, A5B2C19, A5B2C20, A5B2C21, A5B3C1, A5B3C2, A5B3C3, A5B3C4, A5B3C5, A5B3C6, A5B3C7, A5B3C8, A5B3C9, A5B3C10, A5B3C11, A5B3C12, A5B3C13, A5B3C14, A5B3C15, A5B3C16, A5B3C17, A5B3C18, A5B3C19, A5B3C20, A5B3C21, A5B4C1, A5B4C2, A5B4C3, A5B4C4, A5B4C5, A5B4C6, A5B4C7, A5B4C8, A5B4C9, A5B4C10, A5B4C11, A5B4C12, A5B4C13, A5B4C14, A5B4C15, A5B4C16, A5B4C17, A5B4C18, A5B4C19, A5B4C20, A5B4C21, A5B5C1, A5B5C2, A5B5C3, A5B5C4, A5B5C5, A5B5C6, A5B5C7, A5B5C8, A5B5C9, A5B5C10, A5B5C11, A5B5C12, A5B5C13, A5B5C14, A5B5C15, A5B5C16, A5B5C17, A5B5C18, A5B5C19, A5B5C20, A5B5C21, A5B6C1, A5B6C2, A5B6C3, A5B6C4, A5B6C5, A5B6C6, A5B6C7, A5B6C8, A5B6C9, A5B6C10, A5B6C11, A5B6C12, A5B6C13, A5B6C14, A5B6C15, A5B6C16, A5B6C17, A5B6C18, A5B6C19, A5B6C20, A5B6C21, A5B7C1, A5B7C2, A5B7C3, A5B7C4, A5B7C5, A5B7C6, A5B7C7, A5B7C8, A5B7C9, A5B7C10, A5B7C11, A5B7C12, A5B7C13, A5B7C14, A5B7C15, A5B7C16, A5B7C17, A5B7C18, A5B7C19, A5B7C20, A5B7C21, A5B8C1, A5B8C2, A5B8C3, A5B8C4, A5B8C5, A5B8C6, A5B8C7, A5B8C8, A5B8C9, A5B8C10, A5B8C11, A5B8C12, A5B8C13, A5B8C14, A5B8C15, A5B8C16, A5B8C17, A5B8C18, A5B8C19, A5B8C20, A5B8C21, A5B9C1, A5B9C2, A5B9C3, A5B9C4, A5B9C5, A5B9C6, A5B9C7, A5B9C8, A5B9C9, A5B9C10, A5B9C11, A5B9C12, A5B9C13, A5B9C14, A5B9C15, A5B9C16, A5B9C17, A5B9C18, A5B9C19, A5B9C20, A5B9C21, A5B10C1, A5B10C2, A5B10C3, A5B10C4, A5B10C5, A5B10C6, A5B10C7, A5B10C8, A5B10C9, A5B10C10, A5B10C11, A5B10C12, A5B10C13, A5B10C14, A5B10C15, A5B10C16, A5B10C17, A5B10C18, A5B10C19, A5B10C20, A5B10C21, A5B11C1, A5B11C2, A5B11C3, A5B11C4, A5B11C5, A5B11C6, A5B11C7, A5B11C8, A5B11C9, A5B11C10, A5B11C11, A5B11C12, A5B11C13, A5B11C14, A5B11C15, A5B11C16, A5B11C17 , A5B11C18, A5B11C19, A5B11C20, A5B11C21, A5B12C1, A5B12C2, A5B12C3, A5B12C4, A5B12C5, A5B12C6, A5B12C7, A5B12C8, A5B12C9, A5B12C10, A5B12C11, A5B12C12, A5B12C13, A5B12C14, A5B12C15, A5B12C16, A5B12C17, A5B12C18, A5B12C19, A5B12C20, A5B12C21 , A6B1C1, A6B1C2, A6B1C3, A6B1C4, A6B1C5, A6B1C6, A6B1C7, A6B1C8, A6B1C9, A6B1C10, A6B1C11, A6B1C12, A6B1C13, A6B1C14, A6B1C15, A6B1C16, A6B1C17, A6B1C18, A6B1C19, A6B1C20, A6B1C21, A6B2C1, A6B2C2, A6B2C3, A6B2C4 , A6B2C5, A6B2C6, A6B2C7, A6B2C8, A6B2C9, A6B2C10, A6B2C11, A6B2C12, A6B2C13, A6B2C14, A6B2C15, A6B2C16, A6B2C17, A6B2C18, A6B2C19, A6B2C20, A6B2C21, A6B3C1, A6B3C2, A6B3C3, A6B3C4, A6B3C5, A6B3C6, A6B3C7, A6B3C8 , A6B3C9, A6B3C10, A6B3C11, A6B3C12, A6B3C13, A6B3C14, A6B3C15, A6B3C16, A6B3C17, A6B3C18, A6B3C19, A6B3C20, A6B3C21, A6B4C1, A6B4C2, A6B4C3, A6B4C4, A6B4C5, A6B4C6, A6B4C7, A6B4C8, A6B4C9, A6B4C10, A6B4C11, A6B4C12 , A6B4C13, A6B4C14, A6B4C15, A6B4C16, A6B4C17, A6B4C18, A6B4C19, A6B4C20, A6B4C21, A6B5C1, A6B5C2, A6B5C3, A6B5C4, A6B5C 5, A6B5C6, A6B5C7, A6B5C8, A6B5C9, A6B5C10, A6B5C11, A6B5C12, A6B5C13, A6B5C14, A6B5C15, A6B5C16, A6B5C17, A6B5C18, A6B5C19, A6B5C20, A6B5C21, A6B6C1, A6B6C2, A6B6C3, A6B6C4, A6B6C5, A6B6C6, A6B6C7, A6B6C8, A6B6C9, A6B6C10, A6B6C11, A6B6C12, A6B6C13, A6B6C14, A6B6C15, A6B6C16, A6B6C17, A6B6C18, A6B6C19, A6B6C20, A6B6C21, A6B7C1, A6B7C2, A6B7C3, A6B7C4, A6B7C5, A6B7C6, A6B7C7, A6B7C8, A6B7C9, A6B7C10, A6B7C11, A6B7C12, A6B7C13, A6B7C14, A6B7C15, A6B7C16, A6B7C17, A6B7C18, A6B7C19, A6B7C20, A6B7C21, A6B8C1, A6B8C2, A6B8C3, A6B8C4, A6B8C5, A6B8C6, A6B8C7, A6B8C8, A6B8C9, A6B8C10, A6B8C11, A6B8C12, A6B8C13, A6B8C14, A6B8C15, A6B8C16, A6B8C17, A6B8C18, A6B8C19, A6B8C20, A6B8C21, A6B9C1, A6B9C2, A6B9C3, A6B9C4, A6B9C5, A6B9C6, A6B9C7, A6B9C8, A6B9C9, A6B9C10, A6B9C11, A6B9C12, A6B9C13, A6B9C14, A6B9C15, A6B9C16, A6B9C17, A6B9C18, A6B9C19, A6B9C20, A6B9C21, A6B10C1, A6B10C2, A6B10C3, A6B10C4, A6B10C5, A6B10C6, A6B10C7, A6B10C8, A6B10C9, A6B10C10, A6B10C11, A6B10C12, A6B10C13, A6B10C14, A6B10C14 15, A6B10C16, A6B10C17, A6B10C18, A6B10C19, A6B10C20, A6B10C21, A6B11C1, A6B11C2, A6B11C3, A6B11C4, A6B11C5, A6B11C6, A6B11C7, A6B11C8, A6B11C9, A6B11C10, A6B11C11, A6B11C12, A6B11C13, A6B11C14, A6B11C15, A6B11C16, A6B11C17, A6B11C18, A6B11C19, A6B11C20, A6B11C21, A6B12C1, A6B12C2, A6B12C3, A6B12C4, A6B12C5, A6B12C6, A6B12C7, A6B12C8, A6B12C9, A6B12C10, A6B12C11, A6B12C12, A6B12C13, A6B12C14, A6B12C15, A6B12C16, A6B12C17, A6B12C18, A6B12C19, A6B12C20, A6B12C21, A7B1C1, A7B1C2, A7B1C3, A7B1C4, A7B1C5, A7B1C6, A7B1C7, A7B1C8, A7B1C9, A7B1C10, A7B1C11, A7B1C12, A7B1C13, A7B1C14, A7B1C15, A7B1C16, A7B1C17, A7B1C18, A7B1C19, A7B1C20, A7B1C21, A7B2C1, A7B2C2, A7B2C3, A7B2C4, A7B2C5, A7B2C6, A7B2C7, A7B2C8, A7B2C9, A7B2C10, A7B2C11, A7B2C12, A7B2C13, A7B2C14, A7B2C15, A7B2C16, A7B2C17, A7B2C18, A7B2C19, A7B2C20, A7B2C21, A7B3C1, A7B3C2, A7B3C3, A7B3C4, A7B3C5, A7B3C6, A7B3C7, A7B3C8, A7B3C9, A7B3C10, A7B3C11, A7B3C12, A7B3C13, A7B3C14, A7B3C15, A7B3C16, A7B3C17, A7B3C18, A7B3C19, A7B3C20, A7B 3C21, A7B4C1, A7B4C2, A7B4C3, A7B4C4, A7B4C5, A7B4C6, A7B4C7, A7B4C8, A7B4C9, A7B4C10, A7B4C11, A7B4C12, A7B4C13, A7B4C14, A7B4C15, A7B4C16, A7B4C17, A7B4C18, A7B4C19, A7B4C20, A7B4C21, A7B5C1, A7B5C2, A7B5C3, A7B5C4, A7B5C5, A7B5C6, A7B5C7, A7B5C8, A7B5C9, A7B5C10, A7B5C11, A7B5C12, A7B5C13, A7B5C14, A7B5C15, A7B5C16, A7B5C17, A7B5C18, A7B5C19, A7B5C20, A7B5C21, A7B6C1, A7B6C2, A7B6C3, A7B6C4, A7B6C5, A7B6C6, A7B6C7, A7B6C8, A7B6C9, A7B6C10, A7B6C11, A7B6C12, A7B6C13, A7B6C14, A7B6C15, A7B6C16, A7B6C17, A7B6C18, A7B6C19, A7B6C20, A7B6C21, A7B7C1, A7B7C2, A7B7C3, A7B7C4, A7B7C5, A7B7C6, A7B7C7, A7B7C8, A7B7C9, A7B7C10, A7B7C11, A7B7C12, A7B7C13, A7B7C14, A7B7C15, A7B7C16, A7B7C17, A7B7C18, A7B7C19, A7B7C20, A7B7C21, A7B8C1, A7B8C2, A7B8C3, A7B8C4, A7B8C5, A7B8C6, A7B8C7, A7B8C8, A7B8C9, A7B8C10, A7B8C11, A7B8C12, A7B8C13, A7B8C14, A7B8C15, A7B8C16, A7B8C17, A7B8C18, A7B8C19, A7B8C20, A7B8C21, A7B9C1, A7B9C2, A7B9C3, A7B9C4, A7B9C5, A7B9C6, A7B9C7, A7B9C8, A7B9C9, A7B9C10, A7B9C11 C12, A7B9C13, A7B9C14, A7B9C15, A7B9C16, A7B9C17, A7B9C18, A7B9C19, A7B9C20, A7B9C21, A7B10C1, A7B10C2, A7B10C3, A7B10C4, A7B10C5, A7B10C6, A7B10C7, A7B10C8, A7B10C9, A7B10C10, A7B10C11, A7B10C12, A7B10C13, A7B10C14, A7B10C15, A7B10C16, A7B10C17, A7B10C18, A7B10C19, A7B10C20, A7B10C21, A7B11C1, A7B11C2, A7B11C3, A7B11C4, A7B11C5, A7B11C6, A7B11C7, A7B11C8, A7B11C9, A7B11C10, A7B11C11, A7B11C12, A7B11C13, A7B11C14, A7B11C15, A7B11C16, A7B11C17, A7B11C18, A7B11C19, A7B11C20, A7B11C21, A7B12C1, A7B12C2, A7B12C3, A7B12C4, A7B12C5, A7B12C6, A7B12C7, A7B12C8, A7B12C9, A7B12C10, A7B12C11, A7B12C12, A7B12C13, A7B12C14, A7B12C15, A7B12C16, A7B12C17, A7B12C18, A7B12C19, A7B12C20, A7B12C21, A8B1C1, A8B1C2, A8B1C3, A8B1C4, A8B1C5, A8B1C6, A8B1C7, A8B1C8, A8B1C9, A8B1C10, A8B1C11, A8B1C12, A8B1C13, A8B1C14, A8B1C15, A8B1C16, A8B1C17, A8B1C18, A8B1C19, A8B1C20, A8B1C21, A8B2C1, A8B2C2, A8B2C3, A8B2C4, A8B2C5, A8B2C6, A8B2C7, A8B2C8, A8B2C9, A8B2C10, A8B2C11, A8B2C12, A8B2C13, A8B2C14, A8B2C15, A8B2C 16, A8B2C17, A8B2C18, A8B2C19, A8B2C20, A8B2C21, A8B3C1, A8B3C2, A8B3C3, A8B3C4, A8B3C5, A8B3C6, A8B3C7, A8B3C8, A8B3C9, A8B3C10, A8B3C11, A8B3C12, A8B3C13, A8B3C14, A8B3C15, A8B3C16, A8B3C17, A8B3C18, A8B3C19, A8B3C20, A8B3C21, A8B4C1, A8B4C2, A8B4C3, A8B4C4, A8B4C5, A8B4C6, A8B4C7, A8B4C8, A8B4C9, A8B4C10, A8B4C11, A8B4C12, A8B4C13, A8B4C14, A8B4C15, A8B4C16, A8B4C17, A8B4C18, A8B4C19, A8B4C20, A8B4C21, A8B5C1, A8B5C2, A8B5C3, A8B5C4, A8B5C5, A8B5C6, A8B5C7, A8B5C8, A8B5C9, A8B5C10, A8B5C11, A8B5C12, A8B5C13, A8B5C14, A8B5C15, A8B5C16, A8B5C17, A8B5C18, A8B5C19, A8B5C20, A8B5C21, A8B6C1, A8B6C2, A8B6C3, A8B6C4, A8B6C5, A8B6C6, A8B6C7, A8B6C8, A8B6C9, A8B6C10, A8B6C11, A8B6C12, A8B6C13, A8B6C14, A8B6C15, A8B6C16, A8B6C17, A8B6C18, A8B6C19, A8B6C20, A8B6C21, A8B7C1, A8B7C2, A8B7C3, A8B7C4, A8B7C5, A8B7C6, A8B7C7, A8B7C8, A8B7C9, A8B7C10, A8B7C11 A8B7C12 A8B7C13 A8B7C14 A8B7C15 A8B7C16 A8B7C17 A8B7C18 A8B7C19 A8B7C20 A8B7C21 A8B8C1 A8B8C2 A8B8C3 A8B8C4 A8B8C5 A8B 8C7, A8B8C8, A8B8C9, A8B8C10, A8B8C11, A8B8C12, A8B8C13, A8B8C14, A8B8C15, A8B8C16, A8B8C17, A8B8C18, A8B8C19, A8B8C20, A8B8C21, A8B9C1, A8B9C2, A8B9C3, A8B9C4, A8B9C5, A8B9C6, A8B9C7, A8B9C8, A8B9C9, A8B9C10, A8B9C11, A8B9C12, A8B9C13, A8B9C14, A8B9C15, A8B9C16, A8B9C17, A8B9C18, A8B9C19, A8B9C20, A8B9C21, A8B10C1, A8B10C2, A8B10C3, A8B10C4, A8B10C5, A8B10C6, A8B10C7, A8B10C8, A8B10C9, A8B10C10, A8B10C11, A8B10C12, A8B10C13, A8B10C14, A8B10C15, A8B10C16, A8B10C17, A8B10C18, A8B10C19, A8B10C20, A8B10C21, A8B11C1, A8B11C2, A8B11C3, A8B11C4, A8B11C5, A8B11C6, A8B11C7, A8B11C8, A8B11C9, A8B11C10, A8B11C11, A8B11C12, A8B11C13, A8B11C14, A8B11C15, A8B11C16, A8B11C17, A8B11C18, A8B11C19, A8B11C20, A8B11C21, A8B12C1, A8B12C2, A8B12C3, A8B12C4, A8B12C5, A8B12C6, A8B12C7, A8B12C8, A8B12C9, A8B12C10, A8B12C11, A8B12C12, A8B12C13, A8B12C14, A8B12C15, A8B12C16, A8B12C17, A8B12C18, A8B12C19, A8B12C20, A8B12C21, A9B1C1, A9B1C2, A9B1C3, A9B1C4, A9B1C5, A9B1C6, A9B1C7, A9B1C8, A9B1C9, A9B1C10, A9B1C11 , A9B1C12, A9B1C13, A9B1C14, A9B1C15, A9B1C16, A9B1C17, A9B1C18, A9B1C19, A9B1C20, A9B1C21, A9B2C1, A9B2C2, A9B2C3, A9B2C4, A9B2C5, A9B2C6, A9B2C7, A9B2C8, A9B2C9, A9B2C10, A9B2C11, A9B2C12, A9B2C13, A9B2C14, A9B2C15 , A9B2C16, A9B2C17, A9B2C18, A9B2C19, A9B2C20, A9B2C21, A9B3C1, A9B3C2, A9B3C3, A9B3C4, A9B3C5, A9B3C6, A9B3C7, A9B3C8, A9B3C9, A9B3C10, A9B3C11, A9B3C12, A9B3C13, A9B3C14, A9B3C15, A9B3C16, A9B3C17, A9B3C18, A9B3C19 , A9B3C20, A9B3C21, A9B4C1, A9B4C2, A9B4C3, A9B4C4, A9B4C5, A9B4C6, A9B4C7, A9B4C8, A9B4C9, A9B4C10, A9B4C11, A9B4C12, A9B4C13, A9B4C14, A9B4C15, A9B4C16, A9B4C17, A9B4C18, A9B4C19, A9B4C20, A9B4C21, A9B5C1, A9B5C2 , A9B5C3, A9B5C4, A9B5C5, A9B5C6, A9B5C7, A9B5C8, A9B5C9, A9B5C10, A9B5C11, A9B5C12, A9B5C13, A9B5C14, A9B5C15, A9B5C16, A9B5C17, A9B5C18, A9B5C19, A9B5C20, A9B5C21, A9B6C1, A9B6C2, A9B6C3, A9B6C4, A9B6C5, A9B6C6 A9B6C15, A9B6C16, A9B6C17, A9B6C18, A9B6C19, A9B6C20, A9B6C21, A9B7C1, A9B7C2, A9B7C3, A9B7C4, A9B7C5, A9B7C6, A9B7C7, A9B7C8, A9B7C9, A9B7C10, A9B7C11, A9B7C12, A9B7C13, A9B7C14, A9B7C15, A9B7C16, A9B7C17, A9B7C18, A9B7C19, A9B7C20, A9B7C21, A9B8C1, A9B8C2, A9B8C3, A9B8C4, A9B8C5, A9B8C6, A9B8C7, A9B8C8, A9B8C9, A9B8C10, A9B8C11, A9B8C12, A9B8C13, A9B8C14, A9B8C15, A9B8C16, A9B8C17, A9B8C18, A9B8C19, A9B8C20, A9B8C21, A9B9C1, A9B9C2, A9B9C3, A9B9C4, A9B9C5, A9B9C6, A9B9C7, A9B9C8, A9B9C9, A9B9C10, A9B9C11, A9B9C12, A9B9C13, A9B9C14, A9B9C15, A9B9C16, A9B9C17, A9B9C18, A9B9C19, A9B9C20, A9B9C21, A9B10C1, A9B10C2, A9B10C3, A9B10C4, A9B10C5, A9B10C6, A9B10C7, A9B10C8, A9B10C9, A9B10C10, A9B10C11, A9B10C12, A9B10C13, A9B10C14, A9B10C15, A9B10C16, A9B10C17, A9B10C18, A9B10C19, A9B10C20, A9B10C21, A9B11C1, A9B11C2, A9B11C3, A9B11C4, A9B11C5, A9B11C6, A9B11C7, A9B11C8, A9B11C9, A9B11C10, A9B11C11, A9B11C12, A9B11C13, A9B11C14, A9B11C15, A9B11C16, A9B11C17, A9B11C18, A9B11C19, A9B11C20, A9B11C21, A9B12C1, A9B12C2, A9B12C3, A9B12C4, A9B12C5, A9B12C6, A9B12C7, A9B 12C8, A9B12C9, A9B12C10, A9B12C11, A9B12C12, A9B12C13, A9B12C14, A9B12C15, A9B12C16, A9B12C17, A9B12C18, A9B12C19, A9B12C20, A9B12C21, A10B1C1, A10B1C2, A10B1C3, A10B1C4, A10B1C5, A10B1C6, A10B1C7, A10B1C8, A10B1C9, A10B1C10, A10B1C11, A10B1C12, A10B1C13, A10B1C14, A10B1C15, A10B1C16, A10B1C17, A10B1C18, A10B1C19, A10B1C20, A10B1C21, A10B2C1, A10B2C2, A10B2C3, A10B2C4, A10B2C5, A10B2C6, A10B2C7, A10B2C8, A10B2C9, A10B2C10, A10B2C11, A10B2C12, A10B2C13, A10B2C14, A10B2C15, A10B2C16, A10B2C17, A10B2C18, A10B2C19, A10B2C20, A10B2C21, A10B3C1, A10B3C2, A10B3C3, A10B3C4, A10B3C5, A10B3C6, A10B3C7, A10B3C8, A10B3C9, A10B3C10, A10B3C11, A10B3C12, A10B3C13, A10B3C14, A10B3C15, A10B3C16, A10B3C17, A10B3C18, A10B3C19, A10B3C20, A10B3C21, A10B4C1, A10B4C2, A10B4C3, A10B4C4, A10B4C5, A10B4C6, A10B4C7, A10B4C8, A10B4C9, A10B4C10, A10B4C11, A10B4C12, A10B4C13, A10B4C14, A10B4C15, A10B4C16, A10B4C17, A10B4C18, A10B4C19, A10B4C20, A10B4C21, A10B5C1, A10B5C2, A10B5C3, A10B5C4, A10B5C5, A10B5C6, A10B5C7 , A10B5C8, A10B5C9, A10B5C10, A10B5C11, A10B5C12, A10B5C13, A10B5C14, A10B5C15, A10B5C16, A10B5C17, A10B5C18, A10B5C19, A10B5C20, A10B5C21, A10B6C1, A10B6C2, A10B6C3, A10B6C4, A10B6C5, A10B6C6, A10B6C7, A10B6C8, A10B6C9, A10B6C10, A10B6C11 , A10B6C12, A10B6C13, A10B6C14, A10B6C15, A10B6C16, A10B6C17, A10B6C18, A10B6C19, A10B6C20, A10B6C21, A10B7C1, A10B7C2, A10B7C3, A10B7C4, A10B7C5, A10B7C6, A10B7C7, A10B7C8, A10B7C9, A10B7C10, A10B7C11, A10B7C12, A10B7C13, A10B7C14, A10B7C15 , A10B7C16, A10B7C17, A10B7C18, A10B7C19, A10B7C20, A10B7C21, A10B8C1, A10B8C2, A10B8C3, A10B8C4, A10B8C5, A10B8C6, A10B8C7, A10B8C8, A10B8C9, A10B8C10, A10B8C11, A10B8C12, A10B8C13, A10B8C14, A10B8C15, A10B8C16, A10B8C17, A10B8C18, A10B8C19 , A10B8C20, A10B8C21, A10B9C1, A10B9C2, A10B9C3, A10B9C4, A10B9C5, A10B9C6, A10B9C7, A10B9C8, A10B9C9, A10B9C10, A10B9C11, A10B9C12, A10B9C13, A10B9C14, A10B9C15, A10B9C16, A10B9C17, A10B9C18, A10B9C19, A10B9C20, A10B9C21, A10B10C1, A10B10C2 , A10B10C3, A10B10C4, A10B10C5, A10B10 C6, A10B10C7, A10B10C8, A10B10C9, A10B10C10, A10B10C11, A10B10C12, A10B10C13, A10B10C14, A10B10C15, A10B10C16, A10B10C17, A10B10C18, A10B10C19, A10B10C20, A10B10C21, A10B11C1, A10B11C2, A10B11C3, A10B11C4, A10B11C5, A10B11C6, A10B11C7, A10B11C8, A10B11C9, A10B11C10, A10B11C11, A10B11C12, A10B11C13, A10B11C14, A10B11C15, A10B11C16, A10B11C17, A10B11C18, A10B11C19, A10B11C20, A10B11C21, A10B12C1, A10B12C2, A10B12C3, A10B12C4, A10B12C5, A10B12C6, A10B12C7, A10B12C8, A10B12C9, A10B12C10, A10B12C11, A10B12C12, A10B12C13, A10B12C14, A10B12C15, A10B12C16, A10B12C17, A10B12C18, A10B12C19, A10B12C20, A10B12C21, A11B1C1, A11B1C2, A11B1C3, A11B1C4, A11B1C5, A11B1C6, A11B1C7, A11B1C8, A11B1C9, A11B1C10, A11B1C11, A11B1C12, A11B1C13, A11B1C14, A11B1C15, A11B1C16, A11B1C17, A11B1C18, A11B1C19, A11B1C20, A11B1C21, A11B2C1, A11B2C2, A11B2C3, A11B2C4, A11B2C5, A11B2C6, A11B2C7, A11B2C8, A11B2C9, A11B2C10, A11B2C11, A11B2C12, A11B2C13, A11B2C14, A11B2C15, A11B2C16, A11B2C17, A11B2C18, A11B2C19, A11B2C20 , A11B2C21, A11B3C1, A11B3C2, A11B3C3, A11B3C4, A11B3C5, A11B3C6, A11B3C7, A11B3C8, A11B3C9, A11B3C10, A11B3C11, A11B3C12, A11B3C13, A11B3C14, A11B3C15, A11B3C16, A11B3C17, A11B3C18, A11B3C19, A11B3C20, A11B3C21, A11B4C1, A11B4C2, A11B4C3 , A11B4C4, A11B4C5, A11B4C6, A11B4C7, A11B4C8, A11B4C9, A11B4C10, A11B4C11, A11B4C12, A11B4C13, A11B4C14, A11B4C15, A11B4C16, A11B4C17, A11B4C18, A11B4C19, A11B4C20, A11B4C21, A11B5C1, A11B5C2, A11B5C3, A11B5C4, A11B5C5, A11B5C6, A11B5C7 , A11B5C8, A11B5C9, A11B5C10, A11B5C11, A11B5C12, A11B5C13, A11B5C14, A11B5C15, A11B5C16, A11B5C17, A11B5C18, A11B5C19, A11B5C20, A11B5C21, A11B6C1, A11B6C2, A11B6C3, A11B6C4, A11B6C5, A11B6C6, A11B6C7, A11B6C8, A11B6C9, A11B6C10, A11B6C11 , A11B6C12, A11B6C13, A11B6C14, A11B6C15, A11B6C16, A11B6C17, A11B6C18, A11B6C19, A11B6C20, A11B6C21, A11B7C1, A11B7C2, A11B7C3, A11B7C4, A11B7C5, A11B7C6, A11B7C7, A11B7C8, A11B7C9, A11B7C10, A11B7C11, A11B7C12, A11B7C13, A11B7C14, A11B7C15 , A11B7C16, A11B7C17, A11B7C18, A11B7C19, A11 B7C20, A11B7C21, A11B8C1, A11B8C2, A11B8C3, A11B8C4, A11B8C5, A11B8C6, A11B8C7, A11B8C8, A11B8C9, A11B8C10, A11B8C11, A11B8C12, A11B8C13, A11B8C14, A11B8C15, A11B8C16, A11B8C17, A11B8C18, A11B8C19, A11B8C20, A11B8C21, A11B9C1, A11B9C2, A11B9C3, A11B9C4, A11B9C5, A11B9C6, A11B9C7, A11B9C8, A11B9C9, A11B9C10, A11B9C11, A11B9C12, A11B9C13, A11B9C14, A11B9C15, A11B9C16, A11B9C17, A11B9C18, A11B9C19, A11B9C20, A11B9C21, A11B10C1, A11B10C2, A11B10C3, A11B10C4, A11B10C5, A11B10C6, A11B10C7, A11B10C8, A11B10C9, A11B10C10, A11B10C11, A11B10C12, A11B10C13, A11B10C14, A11B10C15, A11B10C16, A11B10C17, A11B10C18, A11B10C19, A11B10C20, A11B10C21, A11B11C1, A11B11C2, A11B11C3, A11B11C4, A11B11C5, A11B11C6, A11B11C7, A11B11C8, A11B11C9, A11B11C10, A11B11C11, A11B11C12, A11B11C13, A11B11C14, A11B11C15, A11B11C16, A11B11C17, A11B11C18, A11B11C19, A11B11C20, A11B11C21, A11B12C1, A11B12C2, A11B12C3, A11B12C4, A11B12C5, A11B12C6, A11B12C7, A11B12C8, A11B12C9, A11B12C10, A11B12C11, A11B12C12, A11B12C13, A11 B12C14, A11B12C15, A11B12C16, A11B12C17, A11B12C18, A11B12C19, A11B12C20 or A11B12C21.

Antibody molecules

In one embodiment, the antibody molecule binds to PD-1 of a mammal, such as a human. For example, an antibody molecule specifically binds to an epitope, eg, a linear or conformational epitope (eg, an epitope described herein) on PD-1.

As used herein, the term "antibody molecule" refers to a protein, eg, an immunoglobulin chain or fragment thereof, containing at least one immunoglobulin variable domain sequence. The term "antibody molecule" includes, for example, a monoclonal antibody (including a full length antibody that has an immunoglobulin Fc region). In one embodiment, the antibody molecule comprises a full length antibody or a full length immunoglobulin chain. In one embodiment, the antibody molecule comprises an antigen-binding or functional fragment of a full length antibody or a full length immunoglobulin chain. In one embodiment, the antibody molecule is a multispecific antibody molecule, for example, it contains a plurality of immunoglobulin variable domain sequences, where the first immunoglobulin variable domain sequence of such a plurality has a binding specificity for a first epitope, and the second immunoglobulin variable domain sequence of such a plurality has a binding specificity for the second epitope. In one embodiment, the multispecific antibody molecule is a bispecific antibody molecule. A bispecific antibody has specificity for no more than two antigens. A bispecific antibody molecule is characterized by a first immunoglobulin variable domain sequence that has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope.

In an embodiment, the antibody molecule is a monospecific antibody molecule and binds a single epitope. For example, a monospecific antibody molecule having multiple immunoglobulin variable domain sequences, each of which binds the same epitope.

In an embodiment, the antibody molecule is a multispecific antibody molecule, e.g., it comprises 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 an embodiment, the first and second epitopes are on the same antigen, eg, the same protein (or subunit of a multimeric protein). In an embodiment, the first and second epitopes overlap. In an embodiment, the first and second epitopes do not overlap. In an embodiment, the first and second epitopes are on different antigens, eg different proteins (or different subunits of a multimeric protein). In an embodiment, the multispecific antibody molecule comprises a third, fourth, or fifth immunoglobulin variable domain. In an embodiment, the multispecific antibody molecule is a bispecific antibody molecule, a trispecific antibody molecule, or a tetraspecific antibody molecule.

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

In one embodiment, the antibody molecule includes a diabody and a single chain molecule, as well as an antigen-binding antibody fragment (eg, Fab, F(ab') ₂ and Fv). For example, an antibody molecule may include a heavy (H) chain variable domain sequence (abbreviated herein as VH) and a light (L) chain variable domain sequence (abbreviated as VL in the invention). In one embodiment, the antibody molecule comprises or consists of a heavy chain and a light chain (referred to herein as a semi-antibody). In another example, an antibody molecule comprises two heavy (H) chain variable domain sequences and two light (L) chain variable domain sequences, thus forming two antigen-binding sites such as Fab, Fab', F(ab') ₂ , Fc, Fd , Fd', Fv, single chain antibodies (e.g. ScFv), single variable domain antibodies, diabodies (Dab) (bivalent and bispecific) and chimeric (e.g. humanized) antibodies, which can be obtained by modifying whole antibodies, or antibodies, synthesized de novo using recombinant DNA technology. Such functional antibody fragments retain the ability to selectively bind to their respective antigen or receptor. Antibodies and antibody fragments can be from any class of antibodies, including, but not limited to, IgG, IgA, IgM, IgD, and IgE, and any subclass of antibodies (eg, IgG1, IgG2, IgG3, and IgG4). The preparation of antibody molecules can be monoclonal or polyclonal. The antibody molecule can also be a human, humanized, CDR-grafted, or in vitro -produced antibody. The antibody may have a heavy chain constant region selected from, for example, IgG1, IgG2, IgG3, or IgG4. The antibody may also have a light chain selected from, for example, the kappa or lambda chain. The term "immunoglobulin" (Ig) in the present description is used interchangeably with the term "antibody".

Examples of antigen-binding fragments of an antibody molecule include: (i) Fab fragment, a monovalent fragment consisting of VL, VH, CL and CH1 domains; (ii) F(ab') ₂ fragment, bivalent fragment containing two Fab-fragment connected by a disulfide bridge in the hinge region; (iii) an Fd fragment consisting of VH and CH1 domains; (iv) an Fv fragment consisting of the VL and VH domains of one antibody arm; (v) a diabody fragment (dAb) that consists of a VH domain; (vi) a camelid variable domain or a camelized variable domain; (vii) single chain Fv (ScFv), see, for example, Bird et al. (1988) Science 242:423-426; and Huston et al. (1988), Proc. Natl. Acad. sci. USA 85:58795883); (viii) single domain antibody. These antibody fragments are obtained using standard techniques known to those skilled in the art, and these fragments are screened for suitability in the same manner as intact antibodies.

The term "antibody" includes intact molecules as well as their functional fragments. Antibody constant regions can be altered, such as mutated, to alter the properties of the antibody (eg, to increase or decrease one or more of the following: Fc receptor binding, antibody glycosylation, cysteine residue number, effector cell function, or complement function).

The antibody molecules can also be single domain antibodies. Single domain antibodies may include antibodies whose hypervariable regions are part of a single domain polypeptide. Examples include, but are not limited to, heavy chain antibodies, naturally occurring antibodies not originally light chains, single domain antibodies derived from conventional 4 chain antibodies, engineered antibodies, and single domain scaffolds other than scaffolds derived from antibodies. The single domain antibodies can be any antibodies known in the art or any single domain antibodies that may be made in the future. Single domain antibodies can be derived from any species, including but not limited to mice, humans, camels, llamas, fish, sharks, goats, rabbits, and bulls. In accordance with another aspect of the present invention, the single domain antibody is a naturally occurring single domain antibody, known as a heavy chain antibody having no light chains. Such single domain antibodies are disclosed in, for example, WO 9404678. For clarity, such a variable domain derived from a heavy chain antibody that does not naturally have a light chain is referred to herein as a VHH or nanobody so that it can be distinguished from a conventional VH. four chain immunoglobulins. Such a VHH molecule can be obtained from antibodies induced in camelid species ( Camelidae ), such as camels, llamas, dromedaries, alpacas and guanacos. Species other than camelids can produce heavy chain antibodies that do not originally have a light chain; such VHHs are included within the scope of the present invention.

The VH and VL regions are subdivided into regions of hypervariability called "complementarity determining regions" (CDRs) interspersed with regions that are more conserved and called "framework regions" (FRs or FWs).

The extent of the framework and CDRs has been accurately determined using 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 definition used in the Oxford Molecular AbM program for modeling antibodies. See generally, for example, Protein Sequence and Structure Analysis of Antibody Variable Domains, in: Antibody Engineering Lab Manual (Ed.: Duebel, S. and Kontermann, R., Springer-Verlag, Heidelberg).

The terms "complementarity determining region" and "CDR" as used herein refer to amino acid sequences within the variable regions of an antibody that confer antigen specificity and binding affinity. Typically, three CDR regions are present in each heavy chain variable region (HCDR1, HCDR2, HCDR3), and three CDR regions are present in each light chain variable region (LCDR1, LCDR2, LCDR3).

The exact boundaries of the amino acid sequence of each CDR region can be determined using any of a number of well-known schemes, including those described in Kabat et al. (1991), "Sequences of Proteins of Immunological Interest", 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat's numbering scheme"), Al-Lazikani et al., (1997) JMB 273,927-948 ("Chothia's numbering scheme"). As used herein, CDR regions defined according to the Chothia numbering system are also sometimes referred to as "hypervariable loops".

For example, according to Kabat, the CDR amino acid residues in the heavy chain variable region (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable region (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2) and 89-97 (LCDR3). According to Chothia, the CDR amino acid residues in VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3); and amino acid residues in VL are numbered 26-32 (LCDR1), 50-52 (LCDR2) and 91-96 (LCDR3). When combining the CDR definitions of Kabat and Chothia, the CDR regions consist of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2) and 95-102 (HCDR3) in human VH and amino acid residues 24-34 (LCDR1), 50- 56 (LCDR2) and 89-97 (LCDR3) in human VL.

Generally, unless specifically indicated, anti-PD-1 antibody molecules may include any combination of one or more CDRs according to Kabat and/or hypervariable loops according to Chothia, such as those described in Table 1. In one embodiment, the following definitions for antibody molecules are used. against PD-1 described in Table 1: HCDR1 according to the combined definitions of Kabat and Chothia CDRs, and HCCDR 2-3 and LCCDR 1-3 regions according to Kabat's definition of CDRs. By all definitions, each of VH and VL typically includes three CDR regions and four FR regions, N-terminally to C-terminally in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

As used herein, "an immunoglobulin variable domain sequence" refers to an amino acid sequence that can form an immunoglobulin variable domain structure. For example, the sequence may include all or part of the amino acid sequence of a naturally occurring variable domain. For example, the sequence may or may not include one, two or more N- or C-terminal amino acids, or may include other changes that are compatible with the formation of the protein structure.

The term "antigen binding site" refers to the portion of an antibody molecule containing determinants that form a contact surface that binds to a PD-1 polypeptide or epitope thereof. With respect to proteins (or protein mimetics), the antigen binding site typically includes one or more loops (at least four amino acids or amino acid mimetics) forming a contact surface that binds to the PD-1 polypeptide. Typically, the antigen-binding region of an antibody molecule includes at least one or two CDR regions and/or hypervariable loops, or more typically at least three, four, five, or six CDR regions and/or hypervariable loops.

The terms "compete" or "cross-compete" are used interchangeably herein to refer to the ability of an antibody molecule to block the binding of an anti-PD-1 antibody molecule, e.g., an anti-PD-1 antibody molecule of the invention, to a target, e.g., human PD-1. Binding blocking can be direct or indirect (eg, through allosteric modulation of the antibody or target molecule). The extent to which an antibody molecule can block the binding of another antibody molecule to a target, and therefore can be said to compete, can be determined using competitive binding assays, such as FACS assays, ELISA or BIACORE assays. In some embodiments, the implementation of the analysis of competitive binding is a quantitative competitive analysis. In some embodiments, the first anti-PD-1 antibody molecule is said to compete for target binding with the second anti-PD-1 antibody molecule if the binding of the first anti-PD-1 antibody molecule to the target is reduced by 10% or more, for example, by 20% or more, 30% or more 40% or more 50% or more 55% or more 60% or more 65% or more 70% or more 75% or more 80% % or more, 85% or more, 90% or more, 95% or more, 98% or more, 99% or more in a competitive binding assay (eg, in the competitive assay described herein).

The terms "monoclonal antibody" or "monoclonal antibody composition" as used herein refer to a preparation of antibody molecules of a single molecular composition. A monoclonal antibody composition exhibits a single binding specificity and affinity for a particular epitope. Monoclonal antibodies can be produced using hybridoma technology or methods that do not use hybridoma technology (eg, using recombinant methods).

An "effectively human" protein is a protein that does not elicit a neutralizing antibody response, such as a human anti-mouse antibody (HAMA) response. In some cases, HAMA can present a problem, for example, when re-administering an antibody molecule, for example, in the treatment of a chronic or relapsing disease. The HAMA response can render repeated administration of antibodies potentially ineffective due to increased serum antibody clearance (see, for example, Saleh et al., Cancer Immunol. Immunother., 32:180-190 (1990)), as well as due to potential allergic reactions (see ., for example, LoBuglio et al., Hybridoma, 5:5117-5123 (1986)).

The antibody molecule may be a polyclonal or monoclonal antibody. In other embodiments, the implementation of the antibody can be obtained in a recombinant way, for example, obtained using phage display or combinatorial methods.

Phage display methods and combinatorial methods for producing antibodies are known in the art (and are described, for example, in US Pat. (Dower et al.); WO 92/20791 publications (Winter et al.); WO 92/15679 publications (Markland et al.); WO 93/01288 publications (Breitling et al.); WO 92/01047 (McCafferty et al.), WO 92/09690 (Garrard et al.), WO 90/02809 (Ladner et al.), Fuchs et al (1991) Bio/ Technology 9:1370-1372 Hay et al (1992) Hum Antibody Hybridomas 3:81-85 Huse et al (1989) Science 246:1275-1281 Griffths 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 9:1 373-1377; Hoogenboom et al. (1991) Nuc Acid Res 19:4133-4137; and Barbas et al. (1991) PNAS 88:7978-7982, the contents of all of which are incorporated herein by reference).

In one embodiment, the antibody is a fully human antibody (e.g., an antibody produced in mice that have been genetically engineered to produce the antibody from a human immunoglobulin sequence) or a non-human antibody, e.g., a rodent (mouse or rat), goat, primate (e.g. , monkeys), camels. Preferably the non-human antibody is a rodent antibody (mouse or rat antibody). Methods for producing rodent antibodies are known in the art.

Human monoclonal antibodies can be generated using transgenic mice carrying human immunoglobulin genes rather than the mouse system. Splenocytes from such transgenic mice, immunized with an antigen of interest, are used to generate hybridomas that secrete human mAbs with specific affinity for epitopes from human protein (see, for example, International Application Publication WO 91/00906 (Wood et al.), PCT Publication WO 91/10741 (Kucherlapati et al.), International Application Publication WO 92/03918 (Lonberg et al.), International Application Publication 92/03917 (Kay et al.), 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).

The antibody may be an antibody in which the variable region or portion thereof, such as CDR regions, is not created in a human body, such as a rat or mouse. Chimeric, CDR-grafted and humanized antibodies are included within the scope of the present invention. Included within the scope of the present invention are antibodies produced in a non-human body, such as a rat or mouse, and then modified, for example, in a variable framework region or a constant region, to reduce antigenicity in a human body.

Chimeric antibodies can be produced using recombinant DNA technologies known in the art (see International Application Publication PCT/US86/02269 (Robinson et al.); European Patent Application 184,187 (Akira, et al.); European Patent Application 171,496 (Taniguchi, M.); EP 173,494 (Morrison et al.); WO 86/01533 (Neuberger et al.); US 4,816,567 (Cabilly et al.); EP 125,023 (Cabilly et al.); 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).

In a humanized or CDR-grafted antibody, at least one or two, but usually all three, recipient CDR regions (Ig heavy and/or light chains) will be replaced by donor CDR regions. The antibody may be replaced with at least a portion of the non-human CDR region, or only some of the CDR regions may be replaced with non-human CDRs. It is only required to replace some of the CDR regions required for binding of the humanized antibody to PD-1. Preferably, the donor is a rodent antibody, such as a rat or mouse antibody, and the recipient is a human scaffold or a human consensus scaffold. Typically, an immunoglobulin providing CDR regions is referred to as a "donor" and an immunoglobulin providing a scaffold is referred to as an "acceptor". In one embodiment, the donor immunoglobulin is non-human (eg, is a rodent immunoglobulin). The acceptor framework is a natural (eg, human) framework or a consensus framework, or has a sequence that is about 85% or more identical, preferably 90%, 95%, 99% or more identical to them.

As used herein, the term "consensus sequence" refers to a sequence constructed from the most frequently occurring amino acids (or nucleotides) in a family of related sequences (see, for example, Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987). B 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 that family.If two amino acids occur with equal frequency, each can be included in the consensus sequence.The term "consensus framework" refers to the framework region in the consensus sequence of an immunoglobulin .

The antibody can be humanized by methods known in the art (see, for example, Morrison, S. L., 1985, Science 229:1202-1207, Oi et al., 1986, BioTechniques 4:214, and US Patents US 5,585,089, US 5,693,761 and US 5,693,762 (Queen et al.), with the contents of all of these publications incorporated into this application by reference).

Humanized or CDR-grafted antibodies can be obtained by CDR grafting or replacement of CDR regions, where one, two or all of the CDR regions of the immunoglobulin chain can be replaced. See, for example, US 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 US 5,225,539 and the contents of all of these publications are incorporated herein by reference. Winter describes a CDR grafting method that can be used to produce humanized antibodies according to the present invention (UK patent application GB 2188638A, filed March 26, 1987, Winter US 5225539), the contents of which are incorporated herein by reference.

Also included within the scope of the present invention are humanized antibodies in which certain amino acids have been substituted, removed, or added. The criteria for selecting amino acids from a donor are described in US 5,585,089, for example, in sections 12-16 of US 5,585,089, for example, in sections 12-16 of US 5,585,089, the contents of which are incorporated herein by reference. Other methods for humanizing antibodies are described in Padlan et al., EP 519596 A1, published December 23, 1992.

The antibody molecule may be a single chain antibody. The single chain antibody (ScFv) may be engineered (see, for example, Colcher, D. et al. (1999) All. N. Y. Acad. Sci. 880:263-80; and Reiter, Y. (1996) Clin. Cancer Res. 2:245-52). A single chain antibody can be dimerized or multimerized to produce polyvalent antibodies having specificities for different epitopes of the same target protein.

In other embodiments, the antibody molecule has a heavy chain constant region that is selected from, for example, IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE heavy chain constant regions; specifically selected from, for example, the (eg human) IgG1, IgG2, IgG3 and IgG4 heavy chain constant regions. In another embodiment, the antibody molecule has a light chain constant region that is selected, for example, from (eg, human) kappa or lambda light chain constant regions. The constant region can be altered, such as mutated, to modify the properties of the antibody (eg, to increase or decrease one or more of the following properties: Fc receptor binding, antibody glycosylation, number of cysteine residues, effector cell function, and/or complement function ). In one embodiment, the antibody has an effector function and can bind complement. In other embodiments, the antibody does not recruit effector cells or does not bind complement. In another embodiment, the antibody has reduced or no ability to bind to the Fc receptor. For example, it refers to an isotype or subtype, fragment or other mutant that is unable to bind to the Fc receptor, eg, its Fc receptor binding region has been mutated or deleted.

Methods for changing the constant region of an antibody are known in the art. Antibodies with altered function, for example with altered affinity for an effector ligand, such as an FcR on a cell, or the C1 component of complement, can be prepared by replacing at least one amino acid residue in the constant portion of an antibody with another residue (see, for example, EP 388151 A1, US Pat. No. 5,624,821 and US Pat. No. 5,648,260, the contents of which are incorporated herein by reference). A similar type of alteration can be described, the use of which in murine immunoglobulin or immunoglobulin of other species can lead to the reduction or elimination of these functions.

An antibody molecule may be derivatized or attached to another functional molecule (eg, another peptide or protein). As used herein, the term "derivatized" antibody molecule refers to an antibody molecule that has been modified. Derivatization methods include, but are not limited to, attachment of a fluorescent molecule, radionuclide, toxin, enzyme, or affinity ligand such as biotin. Thus, the antibody molecules of the invention are intended to include derivatized and otherwise modified forms of the antibodies described herein, including immunoadhesion molecules. For example, an antibody molecule may be operably linked (by chemical linkage, genetic fusion, non-covalent association, or other means) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or diabody), a detectable, a cytotoxic agent, a pharmaceutical an agent and/or a protein or peptide that can mediate the association of an antibody or antibody portion with another molecule (eg, a streptavidin core region or a polyhistidine tag).

One type of derivatized antibody molecule is obtained by cross-linking two or more antibodies (of the same type or different types, for example, to obtain bispecific antibodies). Suitable crosslinkers include heterobifunctional compounds that have two different reactive groups separated by a suitable spacer (eg m-maleimidobenzoyl-N-hydroxysuccinimide ester) or homobifunctional compounds (eg disuccinimidyl suberate). Such linkers can be obtained from Pierce Chemical Company, Rockford, Ill.

Suitable detectable substances with which to derivatize (or label) an antibody molecule of the invention include fluorescent compounds, various enzymes, prosthetic groups, luminescent materials, bioluminescent materials, fluorescent metal atoms such as europium (Eu) and other lanthanides, and radioactive substances (described below). Examples of fluorescent detectable substances include fluorescein, fluorescein isothiocyanate, rhodamine, 5-dimethylamine-1-chloride-naphthalenesulfonyl chloride, phycoerythrin, and the like. The antibody can also be derivatized with detectable enzymes, such as alkaline phosphatase, horseradish peroxidase (β-galactosidase, acetylcholinesterase, glucose oxidase, etc. When an antibody is derivatized with a detectable enzyme, it is detected by adding additional reagents that are used by the enzyme to obtain a detectable reaction product. For example, if the detectable substance is horseradish peroxidase, the addition of hydrogen peroxide and diaminobenzidine results in a colored reaction product that can be detected.An antibody molecule can also be derivatized with a prosthetic group (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 fi coerythrin; an example of a luminescent material includes luminol; and examples of bioluminescent materials include luciferase, luciferin, and aequorin.

A labeled antibody molecule can be used, for example, for diagnostic and/or experimental purposes in a number of ways, including: (i) isolation of a given antigen using standard techniques such as affinity chromatography or immunoprecipitation; (ii) detecting a given antigen (eg, in a cell lysate or cell supernatant) in order to assess the relative content and expression profile of the protein; (iii) monitoring tissue protein levels as part of a clinical analysis technique, for example, to determine the effectiveness of a given treatment regimen.

Antibody molecules can be conjugated to another molecule, typically a label or therapeutic (eg, cytotoxic or cytostatic) agent or particle. Radioactive isotopes can be used in diagnostic or therapeutic applications.

The invention provides radiolabeled antibody molecules and methods for labeling them. In one embodiment, a method for labeling an antibody molecule is disclosed. The method includes contacting an antibody molecule with a complexing agent, resulting in a conjugated antibody.

As discussed above, an antibody molecule may be conjugated to a therapeutic agent. Therapeutically active radioisotopes have already been indicated. Examples of other therapeutic agents include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracindione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine , tetracaine, lidocaine, propranolol, puromycin, maytansinoids, for example, maytansinol (see US patent 5,208,020), CC-1065 (see US patents 5,475,092, 5,585,499, 5,846,545) and their analogues or homologues. Therapeutic agents include, but are not limited to, antimetabolites (eg, methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil, dacarbazine), alkylating agents (eg, mechlorethamine, thiotepa, chlorambucil, CC-1065, melphalan, carmustine ( BSNU) and lomustine (CCNU), cyclophosphamide, busulfan, dibromomannite, streptozotocin, mitomycin C, and cis-dichlorodiaminplatinum(II) (DDP) cisplatin), anthracyclines (eg, daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (eg, dactinomycin ( formerly actinomycin), bleomycin, mithramycin and anthramycin (AMC)), and antimitotic agents (eg vincristine, vinblastine, taxol and maytansinoids).

In one aspect, the present invention relates to a method for obtaining a target-binding molecule that specifically binds to the target disclosed in this application, for example, the PD-1 receptor. For example, the target-binding molecule is an antibody molecule. The method includes: obtaining a target protein that includes at least a portion of a non-human protein that is homologous (at least 70% identical, 75, 80, 85, 87, 90, 92, 94, 95, 96, 97, 98%) of the corresponding portion of the human target protein, but differs in at least one amino acid (eg, at least one, two, three, four, five, six, seven, eight, or nine amino acids); obtaining an antibody molecule that specifically binds to the antigen; and evaluating the effectiveness of the binding agent in modulating the activity of the target protein. The method may further include administering a binding agent (eg, an antibody molecule) or a derivative thereof (eg, a humanized antibody molecule) to a human.

Molecules of multispecific antibodies

In some embodiments, the antibody molecule is a multispecific (eg, bispecific or trispecific) antibody molecule. Techniques for producing bispecific or heterodimeric antibody molecules are known in the art and include, but are not limited to, for example, the "trough-to-trough" method described, for example, in US5731168; Fc pairing using electrostatic guiding as described, for example, in WO 09/089004, WO 06/106905 and WO 2010/129304; heterodimer formation by SEED (Strand Exchange Engineered Domains - domains constructed by chain exchange), as described, for example, in WO 07/110205; Fab arm replacement as described, for example, in WO 08/119353, WO 2011/131746 and WO 2013/060867; a dual antibody conjugate, for example, cross-linking an antibody to form a bispecific structure using a heterobifunctional reagent having a reactive amino group and a reactive sulfhydryl group, as described, for example, in US4433059; bispecific antibody determinants created by recombination of semi-antibodies (pairs of heavy-light chains or Fab fragments) from different antibodies through a cycle of reduction and oxidation of disulfide bonds between two heavy chains, as described, for example, in US4444878; trifunctional antibodies, eg three Fab' fragments cross-linked via reactive sulfhydryl groups, as described, eg, in US5273743; biosynthetic binding proteins, for example, a pair of scFv fragments linked via C-terminal tails, preferably by chemical crosslinking of a disulfide or a reactive amino group, as described, for example, in US5534254; bifunctional antibodies, eg, Fab fragments with different binding specificities, dimerized by leucine zippers (eg, c-fos and c-jun) that replace the constant domain, as described, eg, in US5582996; bispecific and oligospecific mono- and oligovalent receptors, for example, the VH-CH1 regions of two antibodies (two Fab fragments) connected via a polypeptide spacer between the CH1 region of one antibody and the VH region of the other antibody, usually with associated light chains, as described, for example, in US5591828; bispecific DNA-antibody conjugates, eg cross-linking of antibodies or Fab fragments via a double-stranded DNA fragment, as described, for example, in US5635602; bispecific fusion proteins, eg, an expression construct containing two scFvs with a hydrophilic helical peptide linker between them and a full-length constant region, as described, eg, in US5637481; multivalent and multispecific binding proteins, e.g., a dimer of polypeptides having a first domain with an Ig heavy chain variable region binding region and a second domain with an Ig light chain variable region binding region, commonly referred to as diabodies (higher order structures forming bispecific , trispecific or tetraspecific molecules, as described, for example, in US5837242); minibody constructs with linked VL and VH chains, further connected by peptide spacers to the antibody hinge region and CH3 region, which can dimerize to form bispecific/multivalent molecules, as described, for example, in US5837821; VH and VL domains linked via a short peptide linker (eg, 5 or 10 amino acids) or no linker at all, in any orientation, which can form dimers to form bispecific diabodies; trimers and tetramers, as described, for example, in US5844094; a chain of VH domains (or VL domains, family members) connected by peptide bonds with crosslinkable groups at the C-terminus, additionally linked to VL domains, forming a series of FV fragments (or scFv fragments), as described, for example, in US5864019; and single chain binding polypeptides with a VH domain and a VL domain linked via a peptide linker, which are combined into polyvalent structures by non-covalent or chemical crosslinking to form, for example, homobivalent, heterobivalent, trivalent and tetravalent structures using the ScFv format or the diabody format as described, for example, in US5869620. Дополнительные примеры мультиспецифичных и биспецифичных молекул и способы их получения можно найти, например, в US5910573, US5932448, US5959083, US5989830, US6005079, US6239259, US6294353, US6333396, US6476198, US6511663, US6670453, US6743896, US6809185, US6833441, US7129330, US7183076, US7521056, US7527787, US7534866, US7612181, US2002/004587A1, US2002/076406A1, US2002/103345A1, US2003/207346A1, US2003/211078A1, US2004/219643A1, US2004/220388A1, US2004/242847A1, US2005/003403A1, US2005/004352A1, US2005/069552A1, US2005/079170A1, US2005/100543A1, US2005/136049A1, US2005/136051A1, US2005/163782A1, US2005/266425A1, US2006/083747A1, US2006/120960A1, US2006/204493A1, US2006/263367A1, US2007/004909A1, US2007/087381A1, US2007/ 128150A1, US2007/141049A1, US2007/154901A1, US2007/274985A1, US2008/050370A1, US2008/069820A1, US2008/152645A1, US2008/171855A1, US2008/241884A1, US2008/254512A1, US2008/260738A1, US2009/130106A1, US2009/148905A1, US2009/155275A1, US2009/162359A1, US2009/162360A1, US20 09/175851A1, US2009/175867A1, US2009/232811A1, US2009/234105A1, US2009/263392A1, US2009/274649A1, EP346087A2, WO00/06605A2, WO02/072635A2, WO04/081051A1, WO06/020258A2, WO2007/044887A2, WO2007/095338A2, WO2007/137760A2, WO2008/119353A1, WO2009/021754A2, WO2009/068630A1, WO91/03493A1, Wo93/23537A1, WO94/12625A2, WO95/0917A1, The content of the above applications is fully incorporated into the present description by reference.

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

"Fusion protein" and "fusion polypeptide" refer to a polypeptide having at least two covalently linked parts, where each of the parts is a polypeptide with different properties. The property may be a biological property, such as in vitro or in vivo activity. A property can also be a simple chemical or physical property, such as binding to a target molecule, catalyzing a reaction, etc. These two parts can be connected directly by a single peptide bond or through a peptide linker, but still be in reading frame with each other.

The present invention provides an isolated nucleic acid molecule encoding the above antibody molecule, vectors and their host cells. A nucleic acid molecule includes, without limitation, RNA, genomic DNA, and cDNA.

Examples of agents used in combinations

This application describes methods and compositions that include a combination of one or more of the following: (i) an agent that increases the presentation of an antigen (eg, tumor antigen); (ii) an agent that enhances an effector cell response (eg, activation and/or mobilization of B cells and/or T cells); or (iii) an agent that reduces tumor immunosuppression resulting in treatment of a disorder, eg, a hyperproliferative condition or disorder (eg, cancer). In some embodiments, the combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule as described herein). Examples of agents that can be used in these combinations are provided in this application.

Examples of STING agonists

In an embodiment, the combination includes a STING agonist. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., breast cancer, squamous cell carcinoma, melanoma, ovarian cancer, fallopian tube carcinoma, peritoneal carcinoma, soft tissue sarcoma, cancer esophagus, head and neck cancer, endometrial cancer, cervical cancer, or basal cell carcinoma), such as hemoblastosis (eg, leukemia (eg, chronic lymphocytic leukemia (CLL)) or lymphoma (eg, marginal zone B-cell lymphoma , small cell lymphocytic lymphoma, follicular lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma)). In some embodiments, the cancer is selected from head and neck cancer (e.g., head and neck squamous cell carcinoma (HNSCC), skin cancer (e.g., melanoma), lung cancer (e.g., non-small cell lung cancer (NSCLC)) or breast cancer (e.g., triple negative breast cancer (TNBC)).

In some embodiments, the STING agonist is a cyclic dinucleotide, eg, a cyclic dinucleotide, comprising purine or pyrimidine nucleotide bases (eg, adenosine, guanine, uracil, thymine, or cytosine nucleotide bases). In some embodiments, the cyclic dinucleotide nucleotide bases comprise the same nucleotide base or different nucleotide bases.

In some embodiments, the STING agonist comprises the nucleotide base adenosine or guanosine. In some embodiments, the STING agonist comprises one adenosine nucleotide base and one guanosine nucleotide base. In some embodiments, the STING agonist comprises two adenosine nucleotide bases or two guanosine nucleotide bases.

In some embodiments, the implementation of the STING agonist includes a modified cyclic dinucleotide, for example, including a modified nucleotide base, a modified ribose, or a modified phosphate bond. In some embodiments, the modified cyclic dinucleotide includes a modified phosphate bond, such as a thiophosphate.

In some embodiments, the STING agonist comprises a cyclic dinucleotide (eg, a modified cyclic dinucleotide) with 2', 5' or 3', 5' phosphate bonds. In some embodiments, the STING agonist comprises a cyclic dinucleotide (eg, a modified cyclic dinucleotide) with Rp or Sp stereochemistry around phosphate bonds.

In some embodiments, the STING agonist is Rp,Rp-dithio-2',3'-c-di-AMP (e.g., Rp,Rp-dithio-c-[A(2',5')pA(3',5 ')p]) or its cyclic dinucleotide analogue. In some embodiments, the implementation of the STING agonist is a compound presented in patent publication US2015/0056224 (for example, the compound in Figure 2c, for example, compound 21 or compound 22). In some embodiments, the STING agonist is c-[G(2',5')pG(3',5')p], its O-substituted dithioribose derivative or compound shown in FIG. 4 PCT publications WO 2014/189805 and WO 2014/189806. In some embodiments, the STING agonist is c-[(2',5')pA(3',5')p] or its O-substituted dithioribose derivative, or the compound shown in FIG. 5 PCT publications WO 2014/189805 and WO 2014/189806. In some embodiments, the STING agonist is c-[G(2',5')pA(3',5')p] or a dithioribose O-substituted derivative thereof, or the compound shown in FIG. 5 PCT publications WO 2014/189805 and WO 2014/189806. In some embodiments, the STING agonist is 2'-O-propargyl-cyclic-[A(2',5')pA(3',5')p] (2'-O-propargyl-ML-CDA) or a compound shown in FIG. 7 of PCT publication WO 2014/189806.

Other examples of STING agonists are disclosed, for example, in PCT publications WO 2014/189805 and WO 2014/189806, and U.S. 2015/0056225.

Examples of TLR agonists

In an embodiment, the combination described herein comprises a Toll-like receptor (TLR) agonist. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., breast cancer, squamous cell carcinoma, melanoma, ovarian cancer, fallopian tube carcinoma, peritoneal carcinoma, soft tissue sarcoma, cancer esophagus, head and neck cancer, endometrial cancer, cervical cancer, or basal cell carcinoma), such as hemoblastosis (eg, leukemia (eg, chronic lymphocytic leukemia (CLL)) or lymphoma (eg, marginal zone B-cell lymphoma , small cell lymphocytic lymphoma, follicular lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma)).

TLRs are a family of pattern recognition receptors that were originally identified as sensors in the innate immune system that recognize microbial pathogens. In humans, TLR receptors include TLR-1, TLR-2, TLR-3, TLR-4, TLR-5, TLR-6, TLR-7, TLR-8, TLR-9 and TLR-10. TLR-1, -2, -4, -5 and -6 are expressed on the cell surface, while TLR-3, -7/8 and -9 are expressed in the ER compartment. Subpopulations of human dendritic cells can be identified by specific TLR expression patterns. The myeloid or "normal" subpopulation of human dendritic cells expresses TLR receptors 1-8, while the plasmacytoid subset of human dendritic cells expresses only TLR-7 and TLR-9. Binding of the ligand to TLR receptors activates a cascade of intracellular signaling pathways that induce the production of factors involved in inflammation and immunity. Stimulation of the myeloid subpopulation and the plasmacytoid subpopulation of human dendritic cells results in the priming of antigen-specific CD4+ and CD8+ T cells and activation of NK cells and T cells, respectively.

In some embodiments, the TLR agonist is selected from one or more of a TLR-1 agonist, a TLR-2 agonist, a TLR-3 agonist, a TLR-4 agonist, a TLR-5 agonist, a TLR-6 agonist, a TLR-7 agonist, a TLR- 8, a TLR-9 agonist, a TLR-10 agonist, a TLR-1/2 agonist, a TLR-2/6 agonist, or a TLR-7/8 agonist. In one embodiment, the TLR agonist is a TLR7 agonist.

In some embodiments, the TLR agonist is imiquimod or 3-(2-methylpropyl)-3,5,8-triazatricyclo[7.4.0.0 ^2,6 ]trideca-1(9),2(6),4,7,10, 12-hexaene-7-amine. Imiquimod or 3-(2-methylpropyl)-3,5,8-triazatricyclo[7.4.0.0 ^2,6 ]trideca-1(9),2(6),4,7,10,12-hexaen-7-amine can bind to and activate TLR-7 and/or TLR-8.

In some embodiments, the TLR agonist is 852A. 852A is disclosed, for example, in Inglefield et al. J Interferon Cytokine Res. 2008; 28(4):253-63. 852A can bind to and activate TLR-7 and/or TLR-8.

In some embodiments, the TLR agonist is Bacillus Calmette-Guérin (BCG). BCG can bind to and activate TLR-9.

In some embodiments, the TLR agonist is EMD 120108. EMD 120108 is a synthetic oligonucleotide containing a phosphorothioate oligodeoxynucleotide. EMD 1201081 can bind to and activate TLR-9, for example, in monocytes/macrophages, plasmacytoid dendritic cells (DCs), and B cells, initiating immune signaling pathways, activating B cells, and inducing cytokine production by T helper cells.

In some embodiments, the TLR agonist is IMO-2055. IMO-2055 is a synthetic oligonucleotide containing unmethylated CpG dinucleotides. By mimicking unmethylated CpG sequences in bacterial DNA, IMO-2055 can bind to and activate TLR-9, for example, in monocytes/macrophages, plasmacytoid dendritic cells (DCs), and B cells, initiating immune signaling pathways and activating B cells and DCs, and inducing the production of cytokines by T-helper cells.

Other examples of TLR agonists that can be used in combination include, for example, TLR-1/2 agonists (eg, Pam3Cys), TLR-2 agonists (eg, CFA, MALP2, Pam2Cys, FSL-1, or Hib-OMPC), agonists TLR-3 (e.g., polyriboinosinic:polyribocytidylic acid (poly-I:C), polyadenosine-polyuridylic acid (poly-AU), polyinosine-polycytidylic acid stabilized with poly-L-lysine and carboxymethylcellulose (Hiltonol®)), TLR- 4 (eg, monophosphoryl lipid (MPL), LPS, sialyl-Tn (STn)), TLR-5 agonists (eg, bacterial flagellin), TLR-7 agonists (eg, imiquimod), TLR-7/8 agonists (eg, resiquimod or loxoribin) and TLR-9 agonists (eg unmethylated CpG dinucleotide (CpG-ODN)).

In another embodiment, a TLR agonist is used in combination with a GITR agonist, for example as described in WO2004060319 and International Application Publication WO2014012479.

Examples of VEGFR inhibitors

In one embodiment, the combination described herein comprises a vascular endothelial growth factor (VEGF) receptor inhibitor (eg, an inhibitor of one or more VEGFRs (eg, VEGFR-1, VEGFR-2, VEGFR-3) or VEGF). In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., melanoma, breast cancer, colon cancer, esophageal cancer, gastrointestinal stromal tumor (GIST), cancer kidney (eg, renal cell carcinoma), liver cancer, non-small cell lung cancer (NSCLC), ovarian cancer, pancreatic cancer, prostate cancer, or gastric cancer), such as hemoblastosis (eg, lymphomas).

In some embodiments, the VEGFR inhibitor is vatalanib succinate (Compound A47) or a compound disclosed in EP 296122.

In some embodiment, the VEGFR inhibitor is an inhibitor of one or more of VEGFR-2, PDGFR beta, KIT, or Raf kinase C, 1-methyl-5-((2-(5-(trifluoromethyl)-1H-imidazol-2-yl )pyridin-4-yl)oxy)-N-(4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazol-2-amine (Compound A37) or the compound disclosed in PCT Publication WO 2007/030377.

Other exemplary VEGFR pathway inhibitors that may be used in the combinations disclosed herein include, for example, bevacizumab (Avastin®), axitinib (Inlyta®); brivanib alaninate (BMS-582664, (S)-((R)-1-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[2,1-f][ 1,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate); sorafenib (Nexavar®); pazopanib (Votrient®); sunitinib malate (Sutent®); cediranib (AZD2171, CAS 288383-20-1); vargatef (BIBF1120, CAS 928326-83-4); foretinib (GSK1363089); telatinib (BAY57-9352, CAS 332012-40-5); apatinib (YN968D1, CAS 811803-05-1); imatinib (Glivec®); ponatinib (AP24534, CAS 943319-70-8); tivozanib (AV951, CAS 475108-18-0); regorafenib (BAY73-4506, CAS 755037-03-7); vatalanib dihydrochloride (PTK787, CAS 212141-51-0); brivanib (BMS-540215, CAS 649735-46-6); vandetanib (Caprelsa® or AZD6474); motesaniba diphosphate (AMG706, CAS 857876-30-3, N-(2,3-dihydro-3,3-dimethyl-1H-indol-6-yl)-2-[(4-pyridinylmethyl)amino]-3-pyridinecarboxamide described in PCT publication WO 02/066470); dovitinib lactyl lactic acid (TKI258, CAS 852433-84-2); linfanib (ABT869, CAS 796967-16-3); cabozantinib (XL184, CAS 849217-68-1); lestaurtinib (CAS 111358-88-4); N-[5-[[[5-(1,1-dimethylethyl)-2-oxazolyl]methyl]thio]-2-thiazolyl]-4-piperidinecarboxamide (BMS38703, CAS 345627-80-7); (3R,4R)-4-amino-1-((4-(3-methoxyphenyl)amino)pyrrolo[2,1-f][1,2,4]triazin-5-yl)methyl)piperidin-3- ol (BMS690514); N-(3,4-dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aα,5β,6aα)-octahydro-2-methylcyclopenta[c]pyrrol-5-yl]methoxy]-4 -quinazolinamine (XL647, CAS 781613-23-8); 4-methyl-3-[[1-methyl-6-(3-pyridinyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl]amino]-N-[3-(trifluoromethyl)phenyl]- benzamide (BHG712, CAS 940310-85-0); aflibercept (Eylea®); and endostatin (ENDOSTAR®).

Examples of anti-VEGF antibodies that can be used in the combinations disclosed herein include, for example, a monoclonal antibody that binds to the same epitope as the anti-VEGF monoclonal antibody A4.6.1 produced by ATCC hybridoma HB 10709; recombinant humanized monoclonal antibody against VEGF, created according to Presta et al. (1997) Cancer Res. 57:4593-4599. In one embodiment, the anti-VEGF antibody is bevacizumab (BV), also known as rhuMAb VEGF or Avastin®. It includes mutant human IgG1 framework regions and antigen-binding complementarity-determining regions from the mouse anti-hVEGF monoclonal antibody A.4.6.1, which blocks the binding of human VEGF to its receptors. Bevacizumab and other humanized anti-VEGF antibodies are also described in US Pat. No. 6,884,879, published Feb. 26, 2005. Additional antibodies include G6 or B20 series antibodies (e.g. G6-31, B20-4.1) as described in PCT Publication WO2005/012359, PCT Publication WO2005/044853, the contents of these patent applications are expressly incorporated herein by reference. For additional antibodies, see US Patents 7,060,269, 6,582,959, 6,703,020, 6,054,297, WO98/45332, WO 96/30046, WO94/10202, EP 0666868B1, US Pat. 2003/0190317, 2003/0203409 and 2005/0112126; and also Popkov et al, Journal of Immunological Methods 288: 149-164 (2004). Other antibodies include antibodies that bind to a functional epitope on human VEGF comprising residues F17, M18, D19, Y21, Y25, Q89, 191, K101, E103 and C104, or alternatively including residues F17, Y21, Q22, Y25, D63, 183 and Q89.

Examples of c-MET inhibitors

In one embodiment, the combination described in this application includes a c-MET inhibitor. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., non-small cell lung cancer, pancreatic cancer, liver cancer, thyroid cancer (e.g., anaplastic thyroid carcinoma), tumor brain (e.g., glioblastoma), kidney cancer (e.g., renal cell carcinoma), or head and neck cancer (e.g., head and neck squamous cell carcinoma.) In some embodiments, the cancer is liver cancer, e.g., hepatocellular carcinoma (HCC) ( e.g. c-MET-expressing HCC).

In some embodiments, the c-MET inhibitor is Compound A17 or the compound described in US Pat. Nos. 7,767,675 and 8,420,645). c-MET, a receptor tyrosine kinase overexpressed or mutated in many types of tumor cells, plays an important role in proliferation, survival, invasion, tumor cell metastasis, and tumor angiogenesis. Inhibition of c-MET can cause apoptosis of tumor cells overexpressing c-MET protein or expressing constitutively activated c-MET protein.

In some embodiments, the c-MET inhibitor is JNJ-38877605. JNJ-38877605 is an orally bioavailable small molecule inhibitor of c-Met. JNJ-38877605 binds selectively to c-MET causing inhibition of c-MET phosphorylation and interruption of c-Met signaling pathways.

In some embodiments, the c-MET inhibitor is AMG 208. AMG 208 is a selective small molecule c-MET inhibitor. AMG 208 inhibits ligand-dependent and ligand-independent activation of c-MET by inhibiting its tyrosine kinase activity, which may cause cell growth inhibition in tumors that overexpress c-Met.

In some embodiments, the c-MET inhibitor is AMG 337. AMG 337 is an orally bioavailable c-Met inhibitor. AMG 337 selectively binds to c-MET causing interruption of c-MET signaling pathways.

In some embodiments, the c-MET inhibitor is LY2801653. LY2801653 is an orally bioavailable small molecule c-Met inhibitor. LY2801653 binds selectively to c-MET causing inhibition of c-MET phosphorylation and interruption of c-Met signaling pathways.

In some embodiments, the c-Met inhibitor is MSC2156119J. MSC2156119J is an orally bioavailable c-Met inhibitor. MSC2156119J binds selectively to c-MET, resulting in inhibition of c-MET phosphorylation and interruption of c-Met-mediated signaling pathways.

In some embodiments, the c-MET inhibitor is capmatinib. Capmatinib is also known as INCB028060. Capmatinib is an orally bioavailable c-MET inhibitor. Capmatinib binds selectively to c-Met, causing inhibition of c-Met phosphorylation and interruption of c-Met signaling pathways.

In some embodiments, the c-MET inhibitor is crizotinib. Crizotinib is also known as PF-02341066. Crizotinib is an orally bioavailable aminopyridine-based inhibitor of anaplastic lymphoma receptor tyrosine kinase (ALK) and c-Met/hepatocyte growth factor receptor (HGFR) kinase. Crizotinib ATP-competitively binds to and inhibits ALK kinase and ALK fusion proteins. In addition, crizotinib inhibits c-Met kinase and interrupts the c-Met signaling pathway. In general, this agent inhibits the growth of tumor cells.

In some embodiments, the c-MET inhibitor is golvatinib. Golvatinib is an orally bioavailable dual inhibitor of c-MET kinase and VEGFR-2 with potential antitumor activity. Golvatinib binds to and inhibits c-MET and VEGFR-2 activity, which may inhibit tumor cell growth and survival of tumor cells that overexpress such receptor tyrosine kinases.

In some embodiments, the c-MET inhibitor is tivantinib. Tivantinib is also known as ARQ 197. Tivantinib is an orally bioavailable small molecule c-MET inhibitor. Tivantinib binds to the c-MET protein and blocks the c-Met signaling pathway, which can induce apoptosis in tumor cells overexpressing the c-MET protein or expressing the constitutively activated c-Met protein.

Examples of TGFb inhibitors

In one embodiment, the combination described herein comprises a transforming growth factor beta (TGF-β) inhibitor. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., brain cancer (e.g., glioma), melanoma, kidney cancer (e.g., renal cell carcinoma), pleural malignant mesothelioma (eg, recurrent pleural malignant mesothelioma) or breast cancer (eg, metastatic breast cancer)). In some embodiments, the cancer is selected from colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS CRC), liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., non-small cell lung cancer (NSCLC)), cancer breast cancer (eg, triple negative breast cancer (TNBC)), TGFb-expressing cancer, pancreatic cancer, prostate cancer, or kidney cancer (eg, renal cell carcinoma).

TGF-β belongs to a large family of structurally similar cytokines, including, for example, bone morphogenetic proteins (BMPs), growth factors and differentiation inducers, activins and inhibins. In some embodiments, the TGF-β inhibitors described herein can bind and/or inhibit one or more TGF-β isoforms (eg, one, two, or all of TGF-β1, TGF-β2, or TGF-β3).

Under normal conditions, TGF-β maintains homeostasis and limits the growth of epithelial, endothelial, neuronal, and hematopoietic cell lines, for example, by inducing antiproliferative and apoptotic responses. Canonical and non-canonical signaling pathways are involved in cellular responses to TGF-β. Activation of the canonical TGF-β/Smad pathway may mediate the antiproliferative effects of TGF-β. The non-canonical TGF-β pathway can activate additional intracellular pathways, e.g., mitogen-activated protein kinases (MAPKs), phosphatidylinositol-3-kinases/protein kinases B, Rho-like GTPases (Tian et al. Cell Signal. 2011; 23(6) :951-62, Blobe et al N Engl J Med 2000;342(18):1350-8), thus modulating epithelial-mesenchymal transition (EMT) and/or cell motility.

Alterations in the TGF-β signaling pathway are associated with human diseases such as cancer, cardiovascular disease, fibrosis, reproductive disorders, and wound healing. Without being limited by theory, it is believed that in some embodiments, the role of TGF-β in cancer depends on some indicators in the disease (eg, tumor stage and genetic change) and/or cellular context. For example, in advanced cancers, TGF-β can modulate the tumor process, for example, by accelerating tumor growth (eg, by inducing EMT), blocking anti-tumor immune responses, increasing tumor-associated fibrosis, or enhancing angiogenesis (Wakefield and Hill Nat Rev Cancer. 2013 ; 13(5):328-41). In some embodiments, a combination comprising a TGF-β inhibitor described herein is used to treat advanced cancer, metastatic cancer, or advanced cancer.

Preclinical results indicate that TGF-β plays an important role in immune regulation (Wojtowicz-Praga Invest New Drugs. 2003; 21(1):21-32; Yang et al. Trends Immunol. 2010; 31(6):220-7 ). TGF-β can lower the host's immune response through several mechanisms, such as rebalancing T helpers to the Th2 immune phenotype; inhibition of the antitumor response of Th1 type and M1-type macrophages; suppression of the functions of cytotoxic CD8+ T-lymphocytes (CTL), NK-lymphocytes and dendritic cells, formation of CD4+CD25+ T-regulatory cells; or activation of M2-type macrophages with pro-tumor activity mediated by the secretion of immunosuppressive cytokines (eg, IL10 or VEGF), pro-inflammatory cytokines (eg, IL6, TNFα, or IL1), and generation of reactive oxygen species (ROS) with genotoxic activity (Yang et al. Trends Immunol 2010; 31(6):220-7; Truty and Urrutia Pancreatology 2007; 7(5-6):423-35; Achyut et al Gastroenterology 2011; 141(4):1167-78).

In some embodiments, the TGF-β inhibitor is fresolimumab (CAS Number: 948564-73-6). Frezolimumab is also known as GC1008. Frezolimumab is a human monoclonal antibody that binds to and inhibits TGF-beta 1, 2, and 3 isoforms.

Тяжелая цепь фрезолимумаба имеет следующую аминокислотную последовательность: QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQGLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRSEDTAVYYCASTLGLVLDAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 238). Fresolimumab light chain has the following amino acid sequence:

ETVLTQSPGTLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQAPRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYADSPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKHKSSVYACENQ3.

Frezolimumab is disclosed, for example, in WO 2006/086469, US 8,383,780 and US 8,591,901.

In some embodiments, the TGF-β inhibitor is XOMA 089. XOMA 089 is also known as XPA.42.089. XOMA 089 is a fully human monoclonal antibody that specifically binds and neutralizes TGF-beta 1 and 2 ligands.

The XOMA 089 heavy chain variable region has the following amino acid sequence:

QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGLWEVRALPSVYWGQGTLVTVSS (SEQ ID NO: 240) (disclosed as SEQ ID NO: 6 in WO 2012/167143). The XOMA 089 light chain variable region has the following amino acid sequence:

SYELTQPPSVSVAPGQTARITCGANDIGSKSVHWYQQKAGQAPVLVVSEDIIRPSGIPERISGSNSGNTATLTISRVEAGDEADYYCQVWDRDSDQYVFGTGTKVTVLG (SEQ ID NO: 241) (disclosed as SEQ ID NO: 8 in WO 2012/167143).

XOMA 089 binds with high affinity to isoforms of human TGF-β. Typically, XOMA 089 binds with high affinity to TGF-β1 and TGF-β2, and to a lesser extent TGF-β3. In BiaCore assays, the K _D of XOMA 089 on human TGF-β is 14.6 pM for TGF-β1, 67.3 pM for TGF-β2 and 948 pM for TGF-β3. Given the high binding affinity for all three TGF-β isoforms, in some embodiments, XOMA 089 is expected to bind to TGF-β1, 2 and 3 at the dose of XOMA 089 described herein. XOMA 089 cross-reacts with rodent and cynomolgus macaque TGF-β and exhibits in vitro and in vivo functional activity, making rodents and cynomolgus monkeys suitable species for toxicological studies.

In some embodiments, the combination includes a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein) and a TGF-β inhibitor (eg, a TGF-β inhibitor described herein).

Without being limited by theory, it is believed that in some embodiments, resistance to PD-1 immunotherapy is associated with the presence of a transcriptional signature that includes, for example, genes associated with TGF-β signaling and TGF-β-dependent processes, such as wound healing or angiogenesis. (Hugo et al. Cell. 2016; 165(1):35-44). In some embodiments, blockade of TGF-β expands the therapeutic window of therapy that inhibits the PD-1/PD-L1 axis. TGF-β inhibitors may interfere with the clinical efficacy of PD-1 immunotherapy, for example, by modulating the tumor microenvironment, such as vasculogenesis, fibrosis, or factors that influence effector T cell recruitment (Yang et al. Trends Immunol. 2010; 31( 6):220-7; Wakefield and Hill Nat Rev Cancer. 2013; 13(5):328-41; Truty and Urrutia Pancreatology. 2007; 7(5-6):423-35).

Without being limited by theory, it is also believed that in some embodiments, a number of elements of the antitumor immunity cycle express both PD-1 and TGF-β receptors, with PD-1 and TGF-β receptors likely to enhance poorly presented cellular signals. For example, in autochthonous prostate cancer models in mice, the use of a dominant negative form of TGFBRII or blocking TGF-β production in T cells delays tumor growth (Donkor et al. Immunity. 2011; 35(1):123-34; Diener et al Lab Invest 2009;89(2):142-51). Studies of transgenic mouse prostate adenocarcinoma (TRAMP) in mice have shown that blocking TGF-β signaling in adoptively transferred T cells increases their persistence and antitumor activity (Chou et al. J Immunol. 2012; 189(8):3936-46) . The antitumor activity of the transferred T cells may gradually decrease, in part due to upregulation of PD-1 in tumor-infiltrating lymphocytes, confirming the efficacy of the combination of PD-1 and TGF-β inhibition as described herein. The use of neutralizing antibodies against PD-1 or against TGF-β can also affect Treg cells, given their high level of PD-1 expression and their sensitivity to TGF-β stimulation (Riella et al. Am J Transplant. 2012; 12 ( 10):2575-87), which confirms the effectiveness of the combination of PD-1 and TGF-β inhibition for cancer treatment, for example, by increasing the modulation of Treg cell differentiation and function.

Without being limited by theory, it is believed that cancerous tumors may use TGF-β to evade immune surveillance, which promotes tumor growth and metastatic progression. For example, in some advanced cancers, high levels of TGF-β are associated with tumor aggressiveness and poor prognosis, and the TGF-β pathway may contribute to one or more of cancer cell motility, invasion, EMT, or stem cell phenotype. Immune regulation mediated by cancer cells and leukocyte populations (eg, through multiple cell-expressed or secreted molecules, eg, IL-10 or TGF-β) may limit response to checkpoint inhibitors as monotherapy in some patients. In some embodiments, combined inhibition of TGF-β with a checkpoint inhibitor (e.g., a PD-1 inhibitor described herein) is used to treat cancer that does not respond or responds poorly to checkpoint inhibitor monotherapy (e.g., an anti-PD-1 antibody). ), for example, pancreatic cancer or colon and rectal cancer (eg, microsatellite stable colon and rectal cancer (MSS-CRC)). In other embodiments, combined inhibition of TGF-β with a checkpoint inhibitor (e.g., a PD-1 inhibitor described herein) is used to treat cancer that exhibits high levels of effector T cell infiltration, e.g., lung cancer (e.g., non-small cell cancer). lung), breast cancer (eg, triple negative breast cancer), liver cancer (eg, hepatocellular carcinoma), prostate cancer, or kidney cancer (eg, clear cell renal cell carcinoma). In some embodiments, the combination of a TGF-β inhibitor and a PD-1 inhibitor results in a synergistic effect.

In one embodiment, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of 0.1 mg/kg to 20 mg/kg, such as 0.1 mg/kg to 15 mg/kg, 0.1 mg/kg to 12 mg/kg, 0.3 mg/kg to 6 mg/kg, between 1 mg/kg and 3 mg/kg, 0.1 mg/kg to 1 mg/kg, 0.1 mg/kg to 0.5 mg/kg, 0.1 mg/kg to 0.3 mg/kg, 0.3 mg/kg to 3 mg/kg, from 0 .3 mg/kg to 1 mg/kg, 3 mg/kg to 6 mg/kg, or 6 mg/kg to 12 mg/kg, for example, at a dose of approximately 0.1 mg/kg, 0.3 mg/kg kg, 0.5 mg/kg, 1 mg/kg, 3 mg/kg, 6 mg/kg, 12 mg/kg or 15 mg/kg, e.g. once a week, once every two weeks, once a three weeks, once every four weeks, or once every six weeks.

In one embodiment, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of 0.1 mg/kg to 15 mg/kg (e.g., 0.3 mg/kg to 12 mg/kg or 1 mg/kg to 6 mg, e.g. approximately 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 6 mg/kg, 12 mg/kg or 15 mg/kg), for example, once every three weeks. For example, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143 may be administered at a dose of 0.1 mg/kg to 1 mg/kg (e.g., 0.1 mg/kg to 1 mg/kg, eg 0.3 mg/kg), eg once every three weeks. In one embodiment, the TGF-β inhibitor XOMA 089 or the compound disclosed in PCT publication WO 2012/167143 is administered intravenously.

In one embodiment, the combination comprises a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, and a PD-1 inhibitor (eg, an anti-PD-1 antibody described herein).

In one embodiment, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of 0.1 mg/kg to 15 mg/kg (e.g., 0.3 mg/kg to 12 mg/kg or 1 mg/kg to 6 mg, e.g. approximately 0.1 mg/kg, 0.3 mg/kg, 1 mg/kg, 3 mg/kg, 6 mg/kg, 12 mg/kg or 15 mg/kg), e.g. once every three weeks, e.g. intravenously, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 50 mg to 500 mg (e.g., 100 mg to 400 mg, eg at a dose of approximately 100 mg, 200 mg, 300 mg or 400 mg), eg once every 3 weeks or once every 4 weeks, eg by intravenous infusion. In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 100 mg to 300 mg (e.g., a dose of about 100 mg, 200 mg, or 300 mg), e.g., once every 3 weeks eg by intravenous infusion.

In some embodiments, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of about 0.1 mg/kg or 0.3 mg/kg, for example, once every 3 weeks, for example, by intravenous infusion, and the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered at a dose of about 100 mg, for example, once every 3 weeks, for example, by intravenous infusion. In some embodiments, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of about 0.3 mg/kg, such as once every 3 weeks, such as by intravenous infusion, and the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered at a dose of about 100 mg or 300 mg, eg, once every 3 weeks, eg, by intravenous infusion. In some embodiments, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT Publication WO 2012/167143, is administered at a dose of about 1 mg/kg, 3 mg/kg, 6 mg/kg, 12 mg/kg, or 15 mg /kg, for example, once every 3 weeks, for example, by intravenous infusion, and the PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of approximately 300 mg, for example, once every 3 weeks, for example, by intravenous infusion.

In some embodiments, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose between 0.1 mg and 0.2 mg (e.g., about 0.1 mg/kg), e.g. , once every three weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 50 mg to 200 mg (for example, about 100 mg), for example, once every three weeks, for example, by intravenous infusion.

In some embodiments, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose between 0.2 mg and 0.5 mg (e.g., about 0.3 mg/kg), e.g. , once every three weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 50 mg to 200 mg (for example, about 100 mg), for example, once every three weeks, for example, by intravenous infusion.

In some embodiments, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose between 0.2 mg and 0.5 mg (e.g., about 0.3 mg/kg), e.g. , once every three weeks, for example by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 400 mg (for example, about 300 mg), for example, once every three weeks, for example, by intravenous infusion.

In some embodiments, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of 0.5 mg to 2 mg (e.g., about 1 mg/kg), e.g., once every three weeks, e.g., by intravenous infusion, and the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose between 200 mg and 400 mg (e.g., about 300 mg), e.g., once every three weeks, e.g. by intravenous infusion.

In some embodiments, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of 2 mg to 5 mg (e.g., about 3 mg/kg), e.g., once every three weeks , for example, by intravenous infusion, and the PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of between 200 mg and 400 mg (for example, approximately 300 mg), for example, once every three weeks, for example, by intravenous infusion.

In some embodiments, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose between 5 mg and 10 mg (e.g., about 6 mg/kg), e.g., once every three weeks , for example, by intravenous infusion, and the PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of between 200 mg and 400 mg (for example, about 300 mg), for example, once every three weeks, for example, by intravenous infusion.

In some embodiments, the TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of 10 mg to 15 mg (e.g., about 12 mg/kg), e.g., once every three weeks , for example, by intravenous infusion, and the PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of between 200 mg and 400 mg (for example, about 300 mg), for example, once every three weeks, for example, by intravenous infusion.

In some embodiments, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered at a dose of 10 mg to 20 mg (e.g., about 15 mg/kg), e.g., once every three weeks , for example, by intravenous infusion, and the PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of between 200 mg and 400 mg (for example, about 300 mg), for example, once every three weeks, for example, by intravenous infusion.

In some embodiments, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered prior to administration of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule). In other embodiments, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is administered after administration of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule). In some embodiments, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) are administered separately, at least 30 minutes apart. for example, at least 1, 1.5 or 2 hours) between two said administrations.

In one embodiment, a TGF-β inhibitor, XOMA 089, or a compound disclosed in PCT publication WO 2012/167143, is used in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat pancreatic cancer, colon cancer and rectum (eg, microsatellite stable colon and rectal cancer (MSS-CRC)), lung cancer (eg, non-small cell lung cancer), breast cancer (eg, triple-negative breast cancer), liver cancer (eg, hepatocellular carcinoma ), prostate cancer, or kidney cancer (eg, clear cell renal cell carcinoma).

Examples of IDO/TDO Inhibitors

In one embodiment, the combination described in this application includes an inhibitor of indolamine-2,3-dioxygenase (IDO) and/or tryptophan-2,3-dioxygenase (TDO). In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., melanoma, non-small cell lung cancer, colon cancer, head and neck squamous cell carcinoma, ovarian cancer, peritoneal cancer, fallopian cancer tubes, breast cancer (for example, metastatic or HER2-negative breast cancer)), for example, hemoblastosis (for example, lymphomas, for example, non-Hodgkin's lymphoma or Hodgkin's lymphoma (for example, diffuse large B-cell lymphoma (DLBCL))).

In some embodiments, the IDO/TDO inhibitor is selected from (4E)-4-[(3-chloro-4-fluoroanilino)nitrosomethylidene]-1,2,5-oxadiazol-3-amine (also known as INCB24360), indoxymod ( 1-methyl-D-tryptophan) or α-cyclohexyl-5H-imidazo[5,1-a]isoindole-5-ethanol (also known as NLG919).

In some embodiments, the IDO/TDO inhibitor is epacadostat (CAS Number: 1204669-58-8). Epacadostat is also known as INCB24360 or INCB024360 (Incyte). Epacadostat is a potent and selective inhibitor of indolamine 2,3-dioxygenase (IDO1) with an IC50 of 10 nM, highly selective for other related enzymes such as IDO2 or tryptophan 2,3-dioxygenase (TDO).

In some embodiments, the IDO/TDO inhibitor is indoxymod (New Link Genetics). Indoxymod, the D-isomer of 1-methyl-tryptophan, is an orally administered small molecule inhibitor of the indolamine 2,3-dioxygenase (IDO) pathway that blocks the mechanisms by which tumors evade immune-mediated destruction.

In some embodiments, the IDO/TDO inhibitor is NLG919 (New Link Genetics). NLG919 is a potent inhibitor of the IDO (indolamine-(2,3)-dioxygenase) pathway with a Ki/EC50 of 7 nM/75 nM in cell-free assays.

In some embodiments, the IDO/TDO inhibitor is F001287 (Flexus/BMS). F001287 is a small molecule inhibitor of indolamine-2,3-dioxygenase 1 (IDO1).

Examples of A2AR Antagonists

In one embodiment, the combination described herein comprises an adenosine A2a receptor (A2aR) antagonist (eg, an A2aR pathway inhibitor, eg, an adenosine inhibitor, eg, an A2aR or CD-73 inhibitor). In some embodiments, the combination is used to treat cancer, such as the cancer described herein. In some embodiments, the cancer is lung cancer, such as non-small cell lung cancer (NSCLC).

In some embodiments, the A2aR antagonist is istradiphyllin (CAS Number: 155270-99-8). Istradephyllin is also known as KW-6002 or 8-[(E)-2-(3,4-dimethoxyphenyl)vinyl]-1,3-diethyl-7-methyl-3,7-dihydro-1H-purine-2,6 -dion. Istradephyllin is disclosed, for example, in LeWitt et al. (2008) Annals of Neurology 63(3): 295-302.

In some embodiments, the A2aR antagonist is tosadenant (Biotie). Tosadenant is also known as SYN115 or 4-hydroxy-N-(4-methoxy-7-morpholin-4-yl-1,3-benzothiazol-2-yl)-4-methylpiperidine-1-carboxamide. Tosadenant blocks the action of endogenous adenosine on the A2a receptor, which leads to the potentiation of the action of dopamine on the D2 receptor and the inhibition of the action of glutamate on the mGluR5 receptor. In some embodiments, the A2aR antagonist is a preladenant (CAS Number: 377727-87-2). The preladenant is also known as SCH 420814 or 2-(2-furanyl)-7-[2-[4-[4-(2-methoxyethoxy)phenyl]-1-piperazinyl]ethyl]7H-pyrazolo[4,3-e ][1,2,4]triazolo[1,5-c]pyrimidine-5-amine. Preladenant was developed as a drug that acted as a potent and selective adenosine A2A receptor antagonist.

In some embodiments, the A2aR antagonist is vipadenan. Vipadenan is also known as BIIB014, V2006 or 3-[(4-amino-3-methylphenyl)methyl]-7-(furan-2-yl)triazolo[4,5-d]pyrimidin-5-amine. In some embodiments, the A2aR antagonist is PBF-509 (Palobiofarma). In some embodiments, an A2aR antagonist, such as PBF-509, is administered at a daily dose of about 80 mg, 160 mg, or 240 mg.

Other examples of A2aR antagonists include, for example, ATL-444, MSX-3, SCH-58261, SCH-412,348, SCH-442,416, VER-6623, VER-6947, VER-7835, CGS-15943 or ZM-241,385.

In some embodiments, the A2aR antagonist is an A2aR pathway antagonist (eg, a CD-73 inhibitor, eg, an anti-CD73 antibody), MEDI9447. MEDI9447 is a monoclonal antibody specific for CD73. Targeting CD73 to extracellular adenosine production may reduce the immunosuppressive effect of adenosine. MEDI9447 has been reported to have a number of activities, for example inhibiting CD73 ectonucleotidase activity, abolishing AMP-mediated lymphocyte suppression and inhibiting syngeneic tumor growth. MEDI9447 can induce changes in myeloid and lymphoid infiltrating leukocyte populations in the tumor microenvironment. Such changes include, for example, an increase in the number of CD8 effector cells and activated macrophages, as well as a decrease in the proportions of myeloid suppressor cells (MSCs) and regulatory T lymphocytes.

Examples of oncolytic viruses

In some embodiments, the implementation of the combination, as described in this application, includes an oncolytic virus. In embodiments, oncolytic viruses are capable of selectively replicating and inducing apoptosis or slowing the growth of a cancer cell. In some cases, oncolytic viruses have no or minimal effect on non-cancerous cells. In some embodiments, the combination is used to treat cancer, such as the cancer described herein. In some embodiments, the cancer is a brain cancer, such as glioblastoma (GBM). An oncolytic virus includes, but is not limited to, oncolytic adenovirus, oncolytic herpes simplex viruses, oncolytic retrovirus, oncolytic parvovirus, oncolytic vaccinia virus, oncolytic Sindbis virus, oncolytic influenza virus, or oncolytic RNA virus (e.g., oncolytic reovirus, oncolytic Newcastle disease virus ( NDV), measles oncolytic virus, or oncolytic vesicular stomatitis virus (VSV)).

In some embodiments, the oncolytic virus is a recombinant herpes virus, such as herpes simplex virus (HSV), that includes a gene encoding GM-CSF. Without being limited by theory, it is believed that in some embodiments, the implementation of the GM-CSF gene can enhance the antitumor immune response, for example, by recruiting and stimulating dendritic cells in the tumor. Other modifications may be made to the virus, for example, to attenuate the virus and/or increase selectivity for cancer cells. In some embodiments, the recombinant herpes virus lacks a functional ICP34.5 gene, a functional ICP47 gene, or both. In some embodiments, the implementation of the deletion of the ICP34.5 gene prevents HSV infection of non-tumor cells or allows tumor-selective replication. In some embodiments, the implementation of the deletion of the ICP47 gene provides or increases the presentation of the antigen. Deletion of ICP47 can cause upregulation of the HSV US11 gene and allows US11 to be expressed as both an immediate early and a non late gene. This may further increase the extent of viral replication and oncolysis of tumor cells. In some embodiments, the implementation of HSV-1 strain JS1 is used, for example, to improve the ability to kill tumor cells compared to other strains of HSV-1. In some embodiments, the recombinant herpes virus is talimogen lagerparepvec (T-VEC). T-VEC is based on herpes simplex virus type 1 (HSV-1) modified to include a gene that codes for human GM-CSF. The talimogen lagerparepvec is described, for example, in WO 2014/036412. In some embodiments, the combination is used to treat melanoma, head and neck cancer, pancreatic cancer, breast cancer, colon and rectal cancer, or kidney cancer (eg, renal cell carcinoma). In some embodiments, the oncolytic virus is a recombinant herpes virus, such as neuroattenuated, replication limited herpes simplex virus type 1 (HSV-1). In some embodiments, the recombinant herpes virus includes a deletion of the ICP34.5 gene. In some embodiments, the implementation of the recombinant herpes virus is not capable of replication in non-dividing cells. In some embodiments, the recombinant herpes virus is constructed from wild-type strain 17. In some embodiments, the recombinant herpes virus is HSV1716 (SEPREHVIR®). Without being limited by theory, it is believed that in some embodiments, upon intratumoral injection, oncolytic HSV1716 replicates and causes lysis of dividing cells, such as tumor cells. HSV1716 is described, for example, in international application publication WO 2003/068809. In some embodiments, the combination is used to treat glioma (e.g., low-grade glioma), head and neck cancer (e.g., head and neck squamous cell carcinoma), melanoma, liver cancer (e.g., hepatocellular carcinoma), pleural mesothelioma, or non-CNS pediatric cancer.

In some embodiments, the oncolytic virus is vaccinia virus. In some embodiments, the vaccinia virus includes a gene that activates the production of cytotoxic T cells (e.g., the TIR-domain-containing, adapter-inducing interferon-β (TRIF) gene), a gene that reduces immune blockade (e.g., the hydroxyprostaglandin- dehydrogenase 15-(NAD) (HPGD)) or both of these genes. In some embodiments, the surface of the vaccinia virus is deglycosylated. In some embodiments, the vaccinia virus has one or more (eg, two, three, or all) properties selected from cancer lysis, immune adaptation, T cell stimulation, or removal of immune inhibition. In some embodiments, the vaccinia virus is constructed from a Western Reserve strain. In some embodiments, the oncolytic virus is WO-12 (Western Oncolytics Ltd., USA). In some embodiments, the combination is used to treat a solid tumor.

In some embodiments, the oncolytic virus is a recombinant polio virus, e.g., a live attenuated, non-pathogenic oncolytic virus, in which the internal ribosome entry site (IRES) is replaced with a heterologous IRES, e.g., to reduce or eliminate neurovirulence. In some embodiments, the heterologous IRES is an IRES from human rhinovirus type 2 (HRV2). In some embodiments, the recombinant polio virus is the oncolytic polio virus PVS-RIPO (also known as PVSRIPO). PVS-RIPO is a genetically modified non-pathogenic variant of the oral poliovirus Sabin type 1 in which the internal ribosome entry site (IRES) is replaced by the IRES from human rhinovirus type 2 (HRV2). Without being limited by theory, it is believed that in some embodiments, PVS-RIPO proliferates selectively in susceptible, non-neuronal cells (eg, glioblastoma multiforme (GBM) cells) due to the presence of heterologous HRV2 in this recombinant virus. Upon intratumoral administration of PVS-RIPO, poliovirus can be selectively taken up by tumor cells and replicated in them, expressing CD155 (the poliovirus receptor, PVR or NECL5), which ultimately leads to tumor cell lysis. CD155, an oncofetal cell adhesion molecule and tumor antigen, is ectopically expressed in some cancers such as GMB and plays a role in tumor cell migration, invasion, and metastasis. PVS-RIPO is described, for example, in international application publication WO 2014/081937. In some embodiments, the combination is used to treat a solid tumor, such as a CD155-expressing solid tumor. In other embodiments, the combination is used to treat brain cancer, such as glioblastoma.

In some embodiments, the oncolytic virus is a virus, such as the recombinant oncolytic virus described in US2010/0178684 A1, which is hereby incorporated by reference in its entirety. In some embodiments, the recombinant oncolytic virus comprises, or includes, a nucleic acid sequence (e.g., a heterologous nucleic acid sequence) encoding an inhibitor of an immune or inflammatory response, e.g., as described in US2010/0178684 A1, incorporated herein by reference in its entirety. In embodiments, the recombinant oncolytic virus, e.g., oncolytic NDV, comprises, or includes, a nucleic acid sequence encoding a pro-apoptotic protein (e.g., apoptin), a cytokine (e.g., GM-CSF, CSF, interferon, interleukin-2 (IL-2) , tumor necrosis factor alpha), an immunoglobulin (eg, an antibody against ED-B fibronectin), a tumor-associated antigen, a bispecific adapter protein (eg, a bispecific antibody or antibody fragment directed against the NDV HN protein and a T-cell costimulatory receptor such as CD3 or CD28; or a fusion protein of human IL-2 and a single chain antibody directed against NDV HN protein). See, for example, Zamarin et al. Future Microbiol. 7.3(2012):347-67, incorporated herein by reference in its entirety. In some embodiments, the oncolytic virus is a chimeric oncolytic NDV described in US 8591881 B2, US 2012/0122185 A1 or US 2014/0271677 A1, which are hereby incorporated by reference in their entirety.

In some embodiments, the oncolytic virus includes a conditionally replicating adenovirus (CRAd) that is designed to replicate exclusively in cancer cells. See, for example, Alemany et al. nature biotechnol. 18(2000):723-27. In some embodiments, the oncolytic adenovirus includes the adenovirus described in Table 1 on page 725 of Alemany et al., incorporated herein by reference in its entirety.

Examples of oncolytic viruses include, but are not limited to, the following:

Group B oncolytic adenovirus (ColoAd1) (PsiOxus Therapeutics Ltd.) (see, for example, clinical trial NCT02053220);

ONCOS-102 (formerly called CGTG-102), which is an adenovirus comprising granulocyte-macrophage colony-stimulating factor (GM-CSF) (Oncos Therapeutics) (see, for example, clinical trial NCT01598129);

VCN-01, which is a genetically modified human oncolytic adenovirus encoding human PH20 hyaluronidase (VCN Biosciences, S.L.) (see, for example, clinical trials NCT02045602 and NCT02045589);

The conditionally replicative adenovirus ICOVIR-5, which is a virus derived from wild-type human adenovirus serotype 5 (Had5) that has been modified to replicate selectively in cancer cells with deregulated retinoblastoma/E2F pathway (Institut Catala d'Oncologia) (see e.g. , clinical trial NCT01864759);

Celyvir, which includes bone marrow autologous mesenchymal stem cells (MSC) infected with the oncolytic adenovirus ICOVIR5 (Hospital Infantil Universitario Nino Jesus, Madrid, Spain/Ramon Alemany) (see, for example, clinical trial NCT01844661);

CG0070, which is a conditionally replicative oncolytic adenovirus serotype 5 (Ad5) in which the human E2F-1 promoter regulates the expression of essential E1a viral genes, thus limiting viral replication and cytotoxicity to Rb pathway-defective tumor cells (Cold Genesys, Inc.) (see, for example, clinical trial NCT02143804); or

DNX-2401 (formerly Delta-24-RGD), which is an adenovirus engineered to replicate selectively in cells deficient in the retinoblastoma (Rb) pathway and more efficiently infect cells that express certain RGD-binding integrins (Clinica Universidad de Navarra, Universidad de Navarra/DNAtrix, Inc.) (see, for example, clinical trial NCT01956734).

In some embodiments, the implementation of the oncolytic virus described in this application is administered by injection, for example, subcutaneous, intra-arterial, intravenous, intramuscular, intrathecal or intraperitoneal injection. In embodiments, the oncolytic virus described herein is administered intratumorally, transdermally, transmucosally, orally, intranasally, or by pulmonary administration.

Examples of vaccines, e.g. scaffold vaccines

In one embodiment, the combination described in this application includes a vaccine, for example, a scaffold vaccine. In some embodiments, the combination is used to treat cancer, such as the cancer described herein.

Cancer vaccines are disclosed, for example, in PCT publications WO 2007/070660 and WO 2012/167230, EP 1960009 B1, US Pat. Components that can be used in cancer vaccines (eg, implantable scaffold materials) are disclosed, for example, in PCT publications WO 2009/102465 and WO 2013/106852. Methods that can be used to administer cancer vaccines are disclosed, for example, in PCT publications WO 2013/158673, WO 2012/048165 and WO 2012/149358.

In some embodiments, the cancer vaccine comprises a macroporous scaffold comprising: (i) a cell or cell recruiting composition, and (ii) an activating signal capable of inducing or stimulating cell migration, and (iii) a bioactive composition applied to or included on/in scaffold that causes modification of cells recruited to the scaffold. Migration of modified cells can be provided by open, interconnected, macropores and an activating signal.

In some embodiments, a cancer vaccine elicits an endogenous immune response against a cancer target upon administration of porous scaffolds carrying a recruiting composition and a target antigen composition, where the endogenous antigen-presenting cell is recruited into the scaffold followed by contact with the antigen, and where said cell remains until the activating signal does not cause the cell to enter the lymph node tissue outside the scaffold, thus stimulating the endogenous immune response against the specified cancer target.

In some embodiments, a cancer vaccine is used to remove a target cell from a mammal using a scaffold composition.

In some embodiments, an in situ cancer vaccine is obtained by recruiting cancer cells to implanted scaffolds and destroying the cells using a cytotoxic agent.

In some embodiments, a cytosine-guanosine oligonucleotide (CpG-ODN) is used as a scaffold component that can effectively reprogram and activate dendritic cells recruited to the scaffold and elicit an effective antitumor response.

In some embodiments, polyinosine-polycytidylic acid (poly-I:C) and/or CpG ODNs are used to act synergistically on tumor inhibition.

In some embodiments, porous rods comprising an immune cell recruiting compound (e.g., GM-CSF) and an immune cell activating compound (e.g., CpG ODN), and optionally including an antigen such as a tumor lysate, are used, for example, to induce an immune response against a vaccine antigen. In some embodiments, pores that promote cell recruitment or exit are formed in situ in the hydrogels following injection of the hydrogel. In some embodiments, an injection porous shape memory hydrogel polymer is used for administration.

In other embodiments, the combinations disclosed herein include a cancer or tumor vaccine. Non-limiting examples of tumor vaccines that may be used include melanoma antigen peptides such as gp100 peptides, MAGE, Trp-2, MART1 and/or tyrosinase antigens, tumor cells transfected to express the cytokine GM-CSF, DNA based vaccines, vaccines based on RNA and vaccine based on viral transduction. A cancer vaccine can be preventive or therapeutic.

Many experimental tumor vaccination strategies have been developed (see Rosenberg, S., 2000, Development of Cancer Vaccines, ASCO Educational Book Spring: 60-62; Logothetis, C., 2000, ASCO Educational Book Spring: 300-302; Khayat, D. 2000, ASCO Educational Book Spring: 414-428 Foon, K. 2000, ASCO Educational Book Spring: 730-738, also see Restifo, N. and Sznol, M., Cancer Vaccines, ch.61, p. 3023-3043, in DeVita, V. et al (eds.), 1997, Cancer: Principles and Practice of Oncology Fifth Edition). According to one of these strategies, the vaccine is obtained using autologous or allogeneic tumor cells. These cell vaccines have been shown to be most effective in transducing tumor cells expressing GM-CSF. GM-CSF has been shown to be a potent antigen presentation activator for tumor vaccination (Dranoff et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90: 3539-43).

Combinations disclosed in this application, for example, blockade of PD-1, can be used in combination with a set of recombinant proteins and/or peptides expressed in a tumor in order to obtain an immune response against these proteins. Such proteins are usually considered by the immune system as its antigens, and therefore it is tolerant towards them. The tumor antigen may also include the protein telomerase, which is required for the synthesis of chromosome telomeres and which is expressed in more than 85% of human cancers and only in a limited number of somatic tissues (Kim, N et al. (1994) Science 266: 2011-2013). (These somatic tissues can be protected from immune attack in a variety of ways.) A tumor antigen can also be "neoantigens" expressed in cancer cells as a result of somatic mutations that alter the sequence of a protein or result in fusion proteins from two unrelated sequences (e.g., bcr-abl on the Philadelphia chromosome), or an idiotype from B-cell tumors .

Other tumor vaccines may include proteins from viruses associated with various forms of human cancer, such as human papillomaviruses (HPV), hepatitis viruses (HBV and HCV), Kaposi's sarcoma herpesvirus (KHSV), and Epstein-Barr virus (EBV). Another form of tumor-specific antigen that can be used in combination with PD-1 blockade is purified heat shock proteins (HSPs) isolated directly from tumor tissue. Such heat shock proteins contain protein fragments from tumor cells, and HSPs are highly efficient in delivering antigen presenting cells to induce antitumor immunity (Suot, R & Srivastava, P (1995) Science 269:1585-1588; Tamura, Y. et al. (1997) Science 278:117-120).

Dendritic cells (DCs) are potent antigen-presenting cells that can be used to prime antigen-specific responses. Dendritic cells can be prepared ex vivo and loaded with various protein and peptide antigens as well as tumor cell extracts (Nestle, F. et al. (1998) Nature Medicine 4:328-332). DCs can also be genetically transduced to express, among other things, such tumor antigens. DCs have also been fused directly to tumor cells for immunization purposes (Kugler, A. et al. (2000) Nature Medicine 6:332-336). As a vaccination method, DC immunization may be effective in combination with another agent, such as PD-1 blockade, to activate more potent antitumor responses.

Examples of Bispecific T Cell Activators

In one embodiment, the combination described in this application includes a bispecific T cell activator. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., gastrointestinal cancer, melanoma, or lung cancer) or hemoblastosis (e.g., lymphoma (e.g., non-Hodgkin's lymphoma) or leukemia (for example, acute lymphoblastic leukemia)).

Bispecific T cell activators (BiTE®) are a class of artificial bispecific monoclonal antibodies that can direct the host's immune system, eg, T cell cytotoxic activity, against cancer cells. Bispecific T cell activators can form a bond between T cells and tumor cells that causes T cells to exhibit cytotoxic activity against tumor cells by producing proteins such as perforin and granzymes, regardless of the presence of MHC I molecules or costimulatory molecules. Such proteins penetrate into tumor cells and initiate cell apoptosis. This activity mimics the physiological processes seen during T-cell attacks on tumor cells.

In some embodiments, the bispecific T cell activator is a fusion protein comprising two single chain variable fragments (scFv) of different antibodies. In some embodiments, one of the scFv fragments binds to T cells, eg via the CD3 receptor, and the other binds to the tumor cell, eg via a tumor-specific molecule.

In some embodiments, the bispecific T cell activator is an NKG2A and CD138 bispecific antibody molecule or a CD3 and TCR bispecific antibody molecule. In some embodiments, the bispecific T cell activator is a bispecific antibody molecule that binds to CD3 and a tumor antigen (eg, EGFR, PSCA, PSMA, EpCAM, HER2, among others).

In some embodiments, the bispecific T cell activator is blinatumomab (CAS Number: 853426-35-4). Blinatumomab is also known as MT103. Blinatumomab specifically targets the CD3 site for T cells and the CD19 site for B cells.

In some embodiments, the bispecific T cell activator is MT110. MT110 is a single chain antibody that specifically interacts with EpCAM and CD3. MT110 is disclosed, for example, in Amann et al. J Immunother. 2009;32(5):452-64.

In some embodiments, the bispecific T cell activator targets melanoma-associated chondroitin sulfate proteoglycan (MCSP). In some embodiments, the bispecific T cell activator targets CD33. In some embodiments, the bispecific T cell activator comprises trastuzumab (targeting HER2/neu), cetuximab, or panitumumab (both targeting the EGF receptor), a functional fragment thereof. In some embodiments, the bispecific T cell activator targets CD66e and EphA2.

An example of a GITR agonist

In one embodiment, the combination described in this application includes a GITR agonist. In some embodiments, the combination is used to treat cancer, such as the cancer described herein, such as a solid tumor or hemoblastosis. In some embodiments, the cancer is selected from forkhead box P3 (FoxP3)-expressing cancer, head and neck cancer (eg, head and neck squamous cell carcinoma (HNSCC)), or lung cancer (eg, non-small cell lung cancer (NSCLC)).

In some embodiments, the GITR agonist is an anti-GITR antibody molecule. Examples of an anti-GITR antibody molecule are described, for example, in WO 2016/057846 (eg Mab7). In some embodiments, the anti-GITR antibody molecule comprises the HCDR1 GFSLSSY (SEQ ID NO: 301) sequence (disclosed as SEQ ID NO: 84 in WO 2016/057846), the HCDR2 WGGGG sequence (SEQ ID NO: 302) (disclosed as SEQ ID NO: 302) : 80 in WO 2016/057846), HCDR3 sequence HAYGHDGGFAMDY (SEQ ID NO: 303) (disclosed as SEQ ID NO: 29 in WO 2016/057846), LCDR1 SESVSSN (SEQ ID NO: 304) (disclosed as SEQ ID NO: 85 in WO 2016/057846), LCDR2 GAS (SEQ ID NO: 305) (disclosed as SEQ ID NO: 82 in WO 2016/057846) and LCDR3 SYSYPF (SEQ ID NO: 306) (disclosed as SEQ ID NO: 83 in WO 2016/057846), all according to Chothia's definition of CDR regions. In other embodiments, the anti-GITR antibody molecule comprises the HCDR1 SYGVD sequence (SEQ ID NO: 307) (disclosed as SEQ ID NO: 22 in WO 2016/057846), the HCDR2 VIWGGGGTYYASSLMG sequence (SEQ ID NO: 308) (disclosed as SEQ ID NO: 308) : 25 in WO 2016/057846), HCDR3 sequence HAYGHDGGFAMDY (SEQ ID NO: 303) (disclosed as SEQ ID NO: 29 in WO 2016/057846), LCDR1 RASESVSSNVA (SEQ ID NO: 309) (disclosed as SEQ ID NO: 30 in WO 2016/057846), LCDR2 GASNRAT (SEQ ID NO: 310) (disclosed as SEQ ID NO: 33 in WO 2016/057846) and LCDR3 GQSYSYPFT (SEQ ID NO: 311) (disclosed as SEQ ID NO: 34 in WO 2016/057846), all according to the definition of Kabat's CDR regions.

In some embodiments, the anti-GITR antibody molecule comprises the heavy chain variable region (VH) amino acid sequence EVQLVESGGGLVQSGGSLRLSCAASGFSLSSYGVDWVRQAPGKGLEWVGVIWGGGGTYYASSLMGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHAYGHDGGFAMDYWGQGTLVTVSS (SEQ ID NO: 312) (disclosed as SEQ ID NO: 312) (disclosed as SEQ ID NO: 312) . In some embodiments, the anti-GITR antibody molecule comprises the light chain variable region (VL) amino acid sequence EIVMTQSPATLSVSPGERATLSCRASESVSSNVAWYQQRPGQAPRLLIYGASNRATGIPARFSGSGSGTDFTLTISRLEPEDFAVYYCGQSYSYPFTFGQGTKLEIK (SEQ ID NO: 313) (disclosed as SEQ ID NO: 7 in WO 2016/057844).

В некоторых вариантах осуществления молекула антитела против GITR включает аминокислотную последовательность тяжелой цепи (HC): EVQLVESGGGLVQSGGSLRLSCAASGFSLSSYGVDWVRQAPGKGLEWVGVIWGGGGTYYASSLMGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHAYGHDGGFAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 314) (раскрытую как SEQ ID NO: 100 в WO 2016/057846). В некоторых вариантах осуществления молекула антитела против GITR включает аминокислотную последовательность легкой цепи (LC) EIVMTQSPATLSVSPGERATLSCRASESVSSNVAWYQQRPGQAPRLLIYGASNRATGIPARFSGSGSGTDFTLTISRLEPEDFAVYYCGQSYSYPFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 315) (раскрытую как SEQ ID NO: 66 в WO 2016/057846).

Examples of GITR agonists include, for example, fusion proteins of GITR and anti-GITR antibodies (e.g. bivalent anti-GITR antibodies), such as the GITR fusion protein described in US Patent: 6,111,090, EP: 0920505B1, US Patent: 8,586,023, PCT Publications: WO 2010/003118 and 2011/090754, or an anti-GITR antibody as described, for example, in US Patent: 7,025,962, EP: 1947183B1, US Patent: 7,812,135, US Patent: 8,388,967, US Patent: 8,591,886, EP Publication: 3EP 19.6 PCT: WO 2011/028683, US Pat. PCT Publications: WO 2007/133822, PCT Publications: WO2005/055808, PCT Publications: WO 99/40196, PCT Publications: WO 2001/03720, PCT Publications: WO99/20758, US Patent: 6,689,607, PCT Publications: WO2006/083289, PCT publications: WO 2005/115451, patent US: 7,618,632, PCT Publications: WO 2011/051726, International Application Publications: WO2004/060319 and International Application Publications: WO2014/012479.

In one embodiment, a GITR agonist is used in combination with a PD-1 inhibitor, for example as described in WO2015/026684.

In another embodiment, a GITR agonist is used in combination with a TLR agonist, for example as described in WO2004/060319 and International Application Publication: WO2014/012479.

Examples of PD-1 inhibitors

PD-1 is a member of the CD28/CTLA-4 family, expressed, for example, on activated CD4+ and CD8+ T cells, T _reg cells and B cells. It negatively regulates signaling and effector T cell function. PD-1 is induced on tumor-infiltrating T cells and can lead to functional debilitation or dysfunction (Keir et al. (2008) Annu. Rev. Immunol. 26:677-704; Pardoll et al. (2012) Nat Rev Cancer 12( 4):252-64). PD-1 delivers a co-inhibitory signal when bound to either of its two ligands, programmed death protein ligand 1 (PD-L1) or programmed death protein ligand 2 (PD-L2). PD-L1 is expressed on a variety of cell types, including T cells, natural killer (NK) cells, macrophages, dendritic cells (DC), B cells, epithelial cells, vascular endothelial cells, and many types of tumors. High expression of PD-L1 in mouse and human tumors is associated with poor clinical prognosis in various forms of cancer ((Keir et al. (2008) Annu. Rev. Immunol. 26:677704; Pardoll et al. (2012) Nat. Rev. Cancer 12(4):252-64) PD-L2 is expressed on dendritic cells, macrophages and some tumors.

For antitumor immunotherapy, a preliminary preclinical and clinical evaluation of blockade of the PD-1 pathway was performed. Blockade of the PD-1 pathway has been shown in both preclinical and clinical studies to restore effector T cell activity and elicit an effective antitumor response. For example, blockade of the PD-1 pathway can restore depleted/impaired effector T cell function (eg, proliferation, IFN-γ secretion, or cytolytic function) and inhibit T _reg cell functions (Keir et al. (2008) Annu. Rev. Immunol 26:677-704; Pardoll et al. (2012) Nat Rev. Cancer 12(4):252-64). Blockade of the PD-1 pathway can be performed by an antibody, an antigen-binding fragment thereof, an immunoadhesin, a fusion protein, or an oligopeptide of PD-1, PD-L1 and/or PD-L2.

As used herein, the term "programmed death protein 1" or "PD-1" includes isoforms of mammalian, e.g. 1. The amino acid sequence of PD-1, eg human PD-1, is known in the art, eg, Shinohara T et al. (1994) Genomics 23(3):704-6; Finger LR, et al. Gene (1997) 197(1-2):177-87.

The anti-PD-1 antibody molecules described herein may be used alone or in combination with one or more additional agents described herein in accordance with the method described herein. In some embodiments, the combinations described herein include a PD-1 inhibitor, for example, an anti-PD-1 antibody molecule (eg, a humanized antibody molecule) as described herein.

In some embodiments, an anti-PD-1 antibody molecule (e.g., an isolated or recombinant antibody molecule) has one or more of the following properties:

(i) binds to PD-1, eg, human PD-1, with high affinity, eg, an affinity constant of at least about 10 ⁷ M ^-1 , typically about 10 ⁸ M ^-1 , and more typically about 10 ⁹ M ^-1 to 10 ¹⁰ M ^-1 or higher;

(ii) does not substantially bind to CD28, CTLA-4, ICOS, or BTLA;

(iii) inhibits or reduces the binding of PD-1 to a PD-1 ligand, eg, PD-L1 or PD-L2, or both;

(iv) binds specifically to an epitope on PD-1E, eg the same or similar epitope as the epitope recognized by the mouse BAP049 monoclonal antibody or the chimeric BAP049 antibody, eg BAP049-chi or BAP049-chi-Y;

(v) exhibits the same or similar binding affinity or specificity, or both, as any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B BAP049 -Clone-C, BAP049-Clone-D or BAP049-Clone-E;

(vi) exhibits the same or similar binding affinity or specificity, or both, as the antibody molecule (eg, heavy chain variable region and light chain variable region) described in Table 1;

(vii) exhibits the same or similar binding affinity or specificity, or both, as an antibody molecule (eg, heavy chain variable region and light chain variable region) having the amino acid sequence shown in Table 1;

(viii) exhibits the same or similar binding affinity or specificity, or both, as an antibody molecule (eg, heavy chain variable region and light chain variable region) encoded by the nucleotide sequence shown in Table 1;

(ix) inhibits, for example competitively inhibits, the binding of the second antibody molecule to PD-1, where the second antibody molecule is an antibody molecule described in this application, for example, an antibody molecule selected from, for example, any of BAP049-hum01, BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 -hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E;

(x) binds the same or overlapping epitope to a second anti-PD-1 antibody molecule, where the second antibody molecule is an antibody molecule described herein, e.g., an antibody molecule selected from, e.g., any of BAP049-hum01, BAP049- hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E;

(xi) competes for binding, and/or binds the same epitope, to a second anti-PD-1 antibody molecule, wherein the second antibody molecule is an antibody molecule described herein, e.g., an antibody molecule selected from, e.g., any of BAP049 -hum01 BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 , BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E;

(xii) has one or more of the biological properties of an antibody molecule described in this application, for example, an antibody molecule selected from, for example, any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049 -hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A -Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E;

(xiii) has one or more pharmacokinetic properties of an antibody molecule described in this application, for example, an antibody molecule selected from, for example, any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049 -hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A -Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E;

(xiv) inhibits one or more activities of PD-1, for example, results in one or more of: an increase in the number of tumor-infiltrating lymphocytes, an increase in T-cell receptor-mediated proliferation, or a decrease in immune escape of cancer cells;

(xv) binds human PD-1 and is cross-reactive with cynomolgus monkey PD-1;

(xvi) binds to one or more residues in the C strand, CC' loop, C' strand, or FG loop of PD-1, or a combination of two, three, or all of the C strand, CC' loop, C' strand, or FG loop of PD-1 , for example, in the analysis of binding using ELISA or BiaCore; or

(xvii) has a VL region that is more conducive to PD-1 binding than a VH region.

In some embodiments, the antibody molecule binds to PD-1 with high affinity, e.g., a K _D that is about the same or at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% higher or lower than the K _D of a mouse or chimeric anti-PD-1 antibody molecule, for example, a mouse or chimeric anti-PD-1 antibody molecule described herein. In some embodiments, the K _D of a mouse or chimeric anti-PD-1 antibody molecule is less than about 0.4, 0.3, 0.2, 0.1, or 0.05 nM, for example, as measured by the BiaCore method. In some embodiments, the K _D of a mouse or chimeric anti-PD-1 antibody molecule is less than about 0.2 nM, such as about 0.135 nM. In other embodiments, the K _D of a mouse or chimeric anti-PD-1 antibody molecule is less than about 10, 5, 3, 2, or 1 nM, such as when measuring binding on cells expressing PD-1 (eg, 300.19 cells). In some embodiments, the K _D of a mouse or chimeric anti-PD-1 antibody molecule is less than about 5 nM, such as about 4.60 nM (or about 0.69 μg/mL).

In some embodiments, an anti-PD-1 antibody molecule binds to PD-1 with a K _off less than 1×10 ^-4 , 5×10 ^-5 , or 1×10 ^-5 s ^-1 , e.g., approximately 1.65×10 ^-5 with ^-1 . In some embodiments, an anti-PD-1 antibody molecule binds to PD-1 with a K _on greater than 1x10 ⁴ , 5x10 ⁴ , 1x10 ⁵ , or 5x10 ⁵ M ^-1 s ^-1 , e.g., about 1, 23×10 ⁵ M ^-1 s ^-1 .

In some embodiments, the level of expression of the antibody molecule is higher, such as at least about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times higher than the level of expression of the mouse or chimeric antibody molecule. , for example, a murine or chimeric anti-PD-1 antibody molecule described herein. In some embodiments, the antibody molecule is expressed in CHO cells.

In some embodiments, an anti-PD-1 antibody molecule reduces one or more PD-1-associated activities with an IC ₅₀ (concentration at 50% inhibition) that is about the same or lower, e.g., at least about 10%, 20 %, 30%, 40%, 50%, 60%, 70%, 80%, or 90% lower than the IC _{50 of} a mouse or chimeric anti-PD-1 antibody molecule, such as the mouse or chimeric anti-PD-1 antibody molecule described in this application. In some embodiments, the IC50 of a mouse or chimeric anti-PD-1 _antibody molecule is less than about 6, 5, 4, 3, 2, or 1 nM, e.g., when measuring binding on cells expressing PD-1 (e.g., 300.19 cells) . In some embodiments, the IC _{50 of} a mouse or chimeric anti-PD-1 antibody molecule is less than about 4 nM, eg, about 3.40 nM (or about 0.51 μg/mL). In some embodiments, the reduced PD-1-associated activity is the binding of PD-L1 and/or PD-L2 to PD-1. In some embodiments, the anti-PD-1 antibody molecule binds to peripheral blood mononuclear cells (PBMCs) activated with staphylococcal enterotoxin B (SEB). In other embodiments, the anti-PD-1 antibody molecule upregulates IL-2 expression in SEB-activated whole blood. For example, an anti-PD-1 antibody increases IL-2 expression by at least about 2, 3, 4, or 5-fold over IL-2 expression when an isotype control (eg, IgG4) is used.

In some embodiments, the anti-PD-1 antibody molecule has increased stability, e.g., is at least about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times more stable in vivo or in vitro than a murine or chimeric anti-PD-1 antibody molecule, for example, a murine or chimeric anti-PD-1 antibody molecule described herein.

In one embodiment, the anti-PD-1 antibody molecule is a humanized antibody molecule and has a 300-700, 400-650, 450-600 T cell epitope analysis based risk score or risk score as described herein.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one antigen-binding region, e.g., a variable region or an antigen-binding fragment thereof, from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 -hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, three, or four variable regions from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049- hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one or two heavy chain variable regions from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one or two light chain variable regions from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, the anti-PD-1 antibody molecule comprises an IgG4 heavy chain constant region, eg, human IgG4. In one embodiment, human IgG4 includes a substitution at position 228 according to EU numbering (eg, substitution Ser for Pro). In another embodiment, the anti-PD-1 antibody molecule comprises an IgG1 heavy chain constant region, eg, human IgG1. In one embodiment, the human IgG1 includes a substitution at position 297 according to EU numbering (eg, substitution of Asn for Ala). In one embodiment, human IgG1 comprises a substitution at position 265 according to EU numbering, a substitution at position 329 according to EU numbering, or both (eg, Asp to Ala at position 265 and Pro to Ala at position 329). In one embodiment, human IgG1 comprises a substitution at position 234 according to the EU numbering, a substitution at position 235 according to the EU numbering, or both of such substitutions (eg, a change from Leu to Ala at position 234 and a change from Leu to Ala at position 235). In one embodiment, the heavy chain constant region comprises the amino acid sequence shown in Table 3, or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98 %, 99% or higher) to her.

In yet another embodiment, the anti-PD-1 antibody molecule comprises a light chain kappa constant region, eg, a human light chain kappa constant region. In one embodiment, the light chain constant region comprises the amino acid sequence shown in Table 3, or a sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98 %, 99% or higher) to her.

In another embodiment, the anti-PD-1 antibody molecule comprises an IgG4 heavy chain constant region, such as human IgG4, and a light chain kappa constant region, such as a human light chain kappa constant region, such as a heavy and light chain constant region, comprising the amino acid sequence shown in Table 3, or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to it. In one embodiment, human IgG4 includes a substitution at position 228 according to EU numbering (eg, substitution Ser for Pro). In yet another embodiment, an anti-PD-1 antibody molecule comprises an IgG1 heavy chain constant region, e.g., human IgG1, and a light chain kappa constant region, e.g., a human light chain kappa constant region, e.g., a heavy and light chain constant region, comprising an amino acid the sequence shown in Table 3, or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to it. In one embodiment, the human IgG1 includes a substitution at position 297 according to EU numbering (eg, substitution of Asn for Ala). In one embodiment, human IgG1 comprises a substitution at position 265 according to EU numbering, a substitution at position 329 according to EU numbering, or both (eg, Asp to Ala at position 265 and Pro to Ala at position 329). In one embodiment, human IgG1 comprises a substitution at position 234 according to the EU numbering, a substitution at position 235 according to the EU numbering, or both of such substitutions (eg, a change from Leu to Ala at position 234 and a change from Leu to Ala at position 235).

In another embodiment, an anti-PD-1 antibody molecule comprises a heavy chain variable domain and constant region, a light chain variable domain and constant region, or both, comprising the amino acid sequence BAP049-Clone-A, BAP049-Clone-B, BAP049- Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences. The PD-1 antibody molecule optionally includes a leader sequence from the heavy chain, light chain, or both, as shown in Table 4; or a sequence substantially identical to them.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three complementarity determining regions (CDRs) from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049- hum01 BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two or three CDR regions (or all CDR regions simultaneously) from a heavy chain variable region comprising the amino acid sequence shown in Table 1 or encoded by a nucleotide sequence, shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid the sequence shown in Table 1 or the encoded nucleotide sequence shown in Table 1.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 -hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions (or all CDR regions simultaneously) from a light chain variable region comprising the amino acid sequence shown in Table 1 or encoded by a nucleotide sequence, shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or encoded by the nucleotide sequence shown in Table 1. In some embodiments, the anti-PD-1 antibody molecule comprises a substitution in the light chain CDR region, e.g., one or more substitutions in CDR1, CDR2, and/or CDR3 of the light chain. chains. In one embodiment, the anti-PD-1 antibody molecule comprises a substitution in the light chain CDR3 at position 102 of the light chain variable region, e.g., a change from a cysteine residue to a tyrosine residue or a cysteine residue to a serine residue, at position 102 of the light chain variable region according to Table 1 ( for example, SEQ ID NO: 16 or 24 for murine or chimeric, unmodified; or any of SEQ ID NO: 34, 42, 46, 54, 58, 62, 66, 70, 74, or 78 for a modified sequence).

In another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, three, four, five, or six CDR regions (or all CDR regions simultaneously) from a heavy and light chain variable region comprising the amino acid sequence shown in Table 1 or encoded by the nucleotide sequence shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, for example, amino acid substitutions or deletions compared to the amino acid sequence shown in Table 1 or the encoded nucleotide sequence shown in Table 1.

In one embodiment, an anti-PD-1 antibody molecule includes all six CDR regions from an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049- hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or those described in Table 1 or encoded by the nucleotide sequence in Table 1 or closely related CDR regions, e.g., CDR regions that are identical or that have at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions , deletions or insertions, such as conservative substitutions). In one embodiment, an anti-PD-1 antibody molecule may include any CDR region described herein. In some embodiments, the anti-PD-1 antibody molecule comprises a substitution in the light chain CDR region, for example, one or more substitutions in the light chain CDR1, CDR2, and/or CDR3. In one embodiment, the anti-PD-1 antibody molecule comprises a substitution in the light chain CDR3 at position 102 of the light chain variable region, e.g., a change from a cysteine residue to a tyrosine residue or a cysteine residue to a serine residue, at position 102 of the light chain variable region according to Table 1 ( for example, SEQ ID NO: 16 or 24 for murine or chimeric, unmodified; or any of SEQ ID NO: 34, 42, 46, 54, 58, 62, 66, 70, 74, or 78 for a modified sequence).

In another embodiment, an anti-PD-1 antibody molecule includes at least one, two, or three CDR regions according to Kabat et al. (e.g., at least one, two, or three CDR regions as defined by Kabat as shown in Table 1) from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049- hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which have at least one amino acid change but no more than two, three or four changes (e.g. substitutions, deletions or insertions, e.g. conservative substitutions) compared to one, two or three CDR regions according to Kabat et al., shown in Table 1.

In another embodiment, an anti-PD-1 antibody molecule includes at least one, two, or three CDR regions according to Kabat et al. (e.g., at least one, two, or three CDR regions as defined by Kabat as shown in Table 1) from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049- hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which have at least one amino acid change but no more than two, three or four changes (e.g. substitutions, deletions or insertions, e.g. conservative substitutions) compared to one, two or three CDR regions according to Kabat et al., shown in Table 1.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, three, four, five, or six CDR regions according to Kabat et al. (e.g., at least one, two, three, four, five, or six CDR regions as defined by Kabat as shown in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10, BAP049-hum11, BAP049-hum12, BAP049-hum13, BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which have at least one amino acid change but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, three, four, five, or six CDR regions according to Kabat et al. shown in Table 1.

In another embodiment, the anti-PD-1 antibody molecule includes all six CDR regions according to Kabat et al. (e.g., all six CDR regions as defined by Kabat as shown in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049- hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which have at least one amino acid change, but no more than two, three, or four changes (e.g., e.g., substitutions, deletions, or insertions, e.g., conservative substitutions), compared to all six CDR regions according to Kabat et al., shown in Table 1. In one embodiment, an anti-PD-1 antibody molecule may include any of the CDR regions described herein.

In another embodiment, the anti-PD-1 antibody molecule comprises at least one, two, or three Chothia hypervariable loops (e.g., at least one, two, or three Chothia hypervariable loops as shown in Table 1) from the heavy chain variable region. an antibody described herein, e.g. an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09 , BAP049-hum10, BAP049-hum11, BAP049-hum12, BAP049-hum13, BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone -D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or at least amino acids from such hypervariable loops that contact PD-1; or which have at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, or three hypervariable loops according to Chothia et al. shown in Table 1.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three Chothia hypervariable loops (e.g., at least one, two, or three Chothia hypervariable loops as shown in Table 1) of the antibody light chain variable region. described in this application, for example, an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B BAP049-Clone-C BAP049-Clone- D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or at least amino acids from such hypervariable loops that contact PD-1; or which have at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, or three hypervariable loops according to Chothia et al. shown in Table 1.

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, three, four, five, or six hypervariable loops (e.g., at least one, two, three, four, five, or six hypervariable loops as defined by Chothia , as shown in Table 1) from the heavy and light chain variable regions of an antibody described herein, for example, an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049 -hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A -Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or at least amino acids from such hypervariable loops that contact PD-1; or which have at least one amino acid change but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, three, four, five, or six hypervariable loops according to Chothia et al. shown in Table 1.

In one embodiment, an anti-PD-1 antibody molecule includes all six hypervariable loops (e.g., all six hypervariable loops as defined by Chothia as shown in Table 1) of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01 BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 -hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E, or closely related hypervariable loops, e.g. hypervariable loops that are identical or that have at least one amino acid change, but no more than two, three or four changes (eg substitutions, deletions or insertions, eg conservative substitutions); or which have at least one amino acid change but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, eg, conservative substitutions) compared to all six hypervariable loops according to Chothia et al. shown in Table 1 In one embodiment, an anti-PD-1 antibody molecule may include any of the hypervariable loops described herein.

In another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three hypervariable loops that have the same canonical structures as the corresponding hypervariable loop of an antibody described herein, e.g., an antibody selected from any of BAP049 -hum01 BAP049-hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 , BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E, for example, the same canonical structures as at least loop 1 and/or loop 2 of the heavy and/or light chain variable domains of an antibody described herein. See, for example, Chothia et al., (1992) J. Mol. Biol. 227:799-817; Tomlinson et al., (1992) J. Mol. Biol. 227:776-798 on descriptions of the canonical structures of hypervariable loops. These structures can be determined by looking at the tables described in the indicated sources.

In some embodiments, an anti-PD-1 antibody molecule comprises a combination of CDR regions or hypervariable loops as defined according to Kabat et al. and Chothia et al.

In one embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions or hypervariable loops from the heavy chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049 -hum02 BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 , BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E, as defined by Kabat and Chothia (e.g., at least one, two or three CDR regions or hypervariable loops as defined by Kabat and Chothia, as shown in Table 1); or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences; or which have at least one amino acid change, but no more than two, three, or four changes (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) compared to one, two, or three CDR regions or hypervariable loops according to Kabat, and /or Chothia shown in Table 1.

For example, an anti-PD-1 antibody molecule may include a VH CDR1 according to Kabat et al. or VH hypervariable loop 1 according to Chothia et al., or combinations thereof, for example as shown in Table 1. In one embodiment, the combination of CDR regions according to Kabat and Chothia CDR1 VH comprises the amino acid sequence GYTFTTYWMH (SEQ ID NO: 224) or the amino acid a sequence substantially identical to it (eg, having at least one amino acid change, but no more than two, three, or four changes (eg, substitutions, deletions, or insertions, eg, conservative substitutions)). The anti-PD-1 antibody molecule may further include, for example, VH CDR regions 2-3 according to Kabat et al. and VL CDR regions 1-3 according to Kabat et al., for example, as shown in Table 1. Thus, in some embodiments, framework regions are defined based on a combination of CDR regions defined according to Kabat et al. and hypervariable loops, defined according to Chothia et al. For example, an anti-PD-1 antibody molecule may include VH FR1 as determined from VH hypervariable loop 1 according to Chothia et al. and VH FR2 defined based on VH CDR regions 1-2 according to Kabat et al., for example, as shown in Table 1. An anti-PD-1 antibody molecule may further include, for example, VH FR regions 3-4, defined on based on VH CDR regions 2-3 according to Kabat et al. and VL 1-4 FR regions defined from VL 1-3 CDRs according to Kabat et al.

An anti-PD-1 antibody molecule may contain any combination of CDR regions or hypervariable loops as defined by Kabat and Chothia. In one embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions from the light chain variable region of an antibody described herein, e.g., an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03 BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E, as defined by Kabat and Chothia (e.g., at least one, two or three CDR regions as defined by Kabat and Chothia, as shown in Table 1).

In an embodiment, e.g., an embodiment including a variable region, a CDR (e.g., a CDR according to Chothia or a CDR according to Kabat), or another sequence as defined herein, e.g., in Table 1, the antibody molecule is a monospecific antibody molecule, a bispecific antibody molecule , or is an antibody molecule that includes an antigen-binding fragment of an antibody, for example, a semi-antibody or an antigen-binding fragment of a semi-antibody. In some embodiments, the antibody molecule is a bispecific antibody molecule having a first binding specificity for PD-1 and a second binding specificity for TIM-3, LAG-3, CEACAM (e.g., CEACAM-1 and/or CEACAM-5), PD -L1 or PD-L2.

In one embodiment, an anti-PD-1 antibody molecule comprises:

(a) a heavy chain variable region (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 light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33;

(b) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 1; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32;

(c) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224, the amino acid sequence of VHCDR2 SEQ ID NO: 5, and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33; or

(d) VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32.

In one embodiment, the PD-1 antibody molecule 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 amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33.

In one embodiment, the PD-1 antibody molecule comprises a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 1; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32.

In one embodiment, the PD-1 antibody molecule comprises a VH comprising the VHCDR1 amino acid sequence of SEQ ID NO: 224, 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 amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33.

In one embodiment, the PD-1 antibody molecule comprises a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32.

In one embodiment, the antibody molecule is a humanized antibody molecule. In another embodiment, the antibody molecule is a monospecific antibody molecule. In yet another embodiment, the antibody molecule is a bispecific antibody molecule.

In one embodiment, an anti-PD-1 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising the VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and

(ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32.

In another embodiment, the anti-PD-1 antibody molecule comprises:

(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: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 5 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and

(ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33.

In one embodiment, the anti-PD-1 antibody molecule comprises the amino acid sequence of VHCDR1 of SEQ ID NO:1. In another embodiment, the anti-PD-1 antibody molecule comprises the amino acid sequence of VHCDR1 of SEQ ID NO:4. In yet another embodiment, the anti-PD-1 antibody molecule comprises the amino acid sequence of VHCDR1 SEQ ID NO: 224.

In one embodiment, the light or heavy chain variable region framework (e.g., the region comprising at least FR1, FR2, FR3, and optionally FR4) of an anti-PD-1 antibody molecule may be selected from: (a) a framework a light or heavy chain variable region region comprising at least 80%, 85%, 87% 90%, 92%, 93%, 95%, 97%, 98%, or preferably 100% of the amino acid residues from the human light variable region framework or a heavy chain, for example, a light or heavy chain variable region framework region residue from a mature human antibody, human germline sequence, or human consensus sequence; (b) a light or heavy chain variable region framework comprising 20% to 80%, 40% to 60%, 60% to 90%, or 70% to 95% of amino acid residues from a human light or heavy chain variable region framework a heavy chain, for example, a light or heavy chain variable region framework region from a mature human antibody, human germline sequence, or human consensus sequence; (c) a non-human framework (eg, a rodent framework); or (d) a non-human framework that has been altered, eg, to remove antigenic or cytotoxic determinants, eg, deimmunized or partially humanized. In one embodiment, a light or heavy chain variable region framework (specifically, FR1, FR2, and/or FR3) comprises a light or heavy chain variable region framework sequence that is at least 70, 75, 80, 85, 87 88, 90, 92, 94, 95, 96, 97, 98, 99% identical to the framework regions of the VL or VH segment of the human germline gene.

In some embodiments, an anti-PD-1 antibody molecule comprises a heavy chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more changes, such as amino acid substitutions or deletions, from the amino acid sequence of BAP049-chi-HC, for example, the amino acid sequence of the FR region in the entire variable region, for example, shown in FIG. 9A-9B, or SEQ ID NOs: 18, 20, 22, or 30. In one embodiment, the anti-PD-1 antibody molecule comprises a heavy chain variable domain having one or more of the following: E at position 1, V at position 5, A at position 9, V in position 11, K in position 12, K in position 13, E in position 16, L in position 18, R in position 19, I or V in position 20, G in position 24, I in position 37, A or S at position 40, T at position 41, S at position 42, R at position 43, M or L at position 48, V or F at position 68, T at position 69, I at position 70, S at position 71 , A or R at position 72, K or N at position 74, T or K at position 76, S or N at position 77, L at position 79, L at position 81, E or Q at position 82, M at position 83 , S or N at position 84, R at position 87, A at position 88 or T at position 91 of the BAP049-chi-HC amino acid sequence, eg, the FR amino acid sequence in the entire variable region, eg, shown in FIG. 9A-9B or SEQ ID NO: 18, 20, 22 or 30.

Alternatively, or in combination with the BAP049-chi-HC heavy chain substitutions described herein, an anti-PD-1 antibody molecule comprises a light chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more amino acid changes, such as amino acid substitutions or deletions, from the BAP049-chi-LC amino acid sequence, such as the amino acid sequence shown in FIG. 10A-10B or SEQ ID NO: 24 or 26. In one embodiment, the anti-PD-1 antibody molecule comprises a heavy chain variable domain having one or more of the following: E at position 1, V at position 2, Q at position 3, L in position 4, T in position 7, D or L or in position 9, F or T in position 10, Q in position 11, S or P in position 12, L or in position 13, S in position 14, P or L or V at position 15, K at position 16, Q or D at position 17, R at position 18, at position 19, S at position 20, I or L at position 21, T at position 22, L at position 43, K at position 48, A or S at position 49, R or Q at position 51, Y at position 55, I at position 64, S or P at position 66, S at position 69, Y at position 73, G at position 74, E at position 76, F at position 79, N at position 82, N at position 83, L or I at position 84, E at position 85, S or P at position 86, D at position 87, A or F or I at position 89, T or Y in position 91, F in position 93 or Y in position 1 02 amino acid sequence of BAP049-chi-LC, such as the amino acid sequence shown in FIG. 10A-10B or SEQ ID NO: 24 or 26.

In other embodiments, the anti-PD-1 antibody molecule comprises one, two, three, or four heavy chain framework regions (e.g., the VHFW amino acid sequence shown in Table 2 or encoded by the nucleotide sequence shown in Table 2) or a sequence substantially identical to them. .

In other embodiments, the anti-PD-1 antibody molecule comprises one, two, three, or four light chain framework regions (e.g., the VLFW amino acid sequence shown in Table 2 or encoded by the nucleotide sequence shown in Table 2) or a sequence substantially identical to them. .

In other embodiments, the anti-PD-1 antibody molecule comprises one, two, three, or four heavy chain framework regions (e.g., the VHFW amino acid sequence shown in Table 2 or encoded by the nucleotide sequence shown in Table 2) or a sequence substantially identical to them. ; and one, two, three, or four light chain framework regions (eg, the VLFW amino acid sequence shown in Table 2 or encoded by the nucleotide sequence shown in Table 2) or a sequence substantially identical to them.

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework region 1 (VHFW1) of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049- hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B BAP049-Clone-C BAP049- Clone-D or BAP049-Clone-E (e.g. SEQ ID NO: 147). In some embodiments, the antibody molecule comprises heavy chain framework 1 (VHFW1) from BAP049-hum14 or BAP049-hum15 (eg, SEQ ID NO: 151).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework region 2 (VHFW2) of BAP049-hum01, BAP049-hum02, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum09, BAP049-hum11, BAP049- hum12, BAP049-hum13, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C or BAP049-Clone-E (e.g. SEQ ID NO: 153). In some embodiments, the antibody molecule comprises heavy chain framework 2 (VHFW2) from BAP049-hum03, BAP049-hum04, BAP049-hum08, BAP049-hum10, BAP049-hum14, BAP049-hum15, or BAP049-Clone-D (e.g., SEQ ID NO: 157). In some embodiments, the antibody molecule comprises heavy chain framework 2 (VHFW2) from BAP049-hum16 (eg, SEQ ID NO: 160).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework region 3 (VHFW3) of BAP049-hum01, BAP049-hum02, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum09, BAP049-hum11, BAP049- hum12, BAP049-hum13, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C or BAP049-Clone-E (e.g. SEQ ID NO: 162). In some embodiments, the antibody molecule comprises heavy chain framework 3 (VHFW3) of BAP049-hum03, BAP049-hum04, BAP049-hum08, BAP049-hum10, BAP049-hum14, BAP049-hum15, BAP049-hum16, or BAP049-Clone-D ( for example, SEQ ID NO: 166).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework 4 (VHFW4) from BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049- hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B BAP049-Clone C, BAP049-Clone-D or BAP049-Clone-E (e.g. SEQ ID NO: 169).

In some embodiments, the anti-PD-1 antibody molecule comprises light chain framework 1 (VLFW1) from BAP049-hum08, BAP049-hum09, BAP049-hum15, BAP049-hum16, or BAP049-Clone-C (e.g., SEQ ID NO: 174) . In some embodiments, the antibody molecule comprises light chain framework 1 (VLFW1) of BAP049-hum01, BAP049-hum04, BAP049-hum05, BAP049-hum07, BAP049-hum10, BAP049-hum11, BAP049-hum14, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-D or BAP049-Clone-E (e.g. SEQ ID NO: 177). In some embodiments, the antibody molecule comprises light chain framework 1 (VLFW1) from BAP049-hum06 (eg, SEQ ID NO: 181). In some embodiments, the antibody molecule comprises light chain framework 1 (VLFW1) from BAP049-hum13 (eg, SEQ ID NO: 183). In some embodiments, the antibody molecule comprises light chain framework 1 (VLFW1) from BAP049-hum02, BAP049-hum03, or BAP049-hum12 (eg, SEQ ID NO: 185).

In some embodiments, the anti-PD-1 antibody molecule comprises light chain framework 2 (VLFW2) of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum06, BAP049-hum08, BAP049-hum09, BAP049-hum10, BAP049- hum11, BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-D, or BAP049-Clone-E (for example, SEQ ID NO: 187). In some embodiments, the antibody molecule comprises light chain framework 2 (VLFW2) from BAP049-hum04, BAP049-hum05, BAP049-hum07, BAP049-hum13, or BAP049-Clone-C (eg, SEQ ID NO: 191). In some embodiments, the antibody molecule comprises light chain framework 2 (VLFW2) from BAP049-hum12 (eg, SEQ ID NO: 194).

In some embodiments, the anti-PD-1 antibody molecule comprises light chain framework 3 (VLFW3) of BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10, BAP049-hum11, BAP049-hum12, BAP049- hum13, BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E (e.g. SEQ ID NO: 196). In some embodiments, the antibody molecule comprises light chain framework 3 (VLFW3) from BAP049-hum02 or BAP049-hum03 (eg, SEQ ID NO: 200). In some embodiments, the antibody molecule comprises light chain framework 3 (VLFW3) from BAP049-hum01 or BAP049-Clone-A (eg, SEQ ID NO: 202). In some embodiments, the antibody molecule comprises light chain framework 3 (VLFW3) from BAP049-hum04, BAP049-hum05, or BAP049-Clone-B (eg, SEQ ID NO: 205).

In some embodiments, the anti-PD-1 antibody molecule comprises light chain framework 4 (VLFW4) of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049- hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B C, BAP049-Clone-D or BAP049-Clone-E (e.g. SEQ ID NO: 208).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 of BAP049-hum01, BAP049-hum02, BAP049-hum05, BAP049-hum06, BAP-hum07, BAP049-hum09, BAP049-hum11, BAP049-hum12 , BAP049-hum13, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C or BAP049-Clone-E (for example, SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2) and SEQ ID NO: 162 (VHFW3)). In some embodiments, the antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum03, BAP049-hum04, BAP049-hum08, BAP049-hum10, or BAP049-Clone-D (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 157 (VHFW2) and SEQ ID NO: 166 (VHFW3)). In some embodiments, the antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum14 or BAP049-hum15 (e.g., SEQ ID NO: 151 (VHFW1), SEQ ID NO: 157 (VHFW2), and SEQ ID NO: 166 (VHFW3 )). In some embodiments, the antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum16 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 160 (VHFW2), and SEQ ID NO: 166 (VHFW3)). In some embodiments, the antibody molecule further comprises a heavy chain framework region 4 (VHFW4) of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049 -hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B BAP049-Clone-C BAP049 -Clone-D or BAP049-Clone-E (eg SEQ ID NO: 169).

In some embodiments, the anti-PD-1 antibody molecule comprises light chain framework regions 1-3 from BAP049-hum01 or BAP049-Clone-A (e.g., SEQ ID NO: 177 (VLFW1), SEQ ID NO: 187 (VLFW2), and SEQ ID NO: 202 (VLFW3)). In some embodiments, the antibody molecule comprises light chain framework regions 1-3 from BAP049-hum02 or BAP049-hum03 (e.g., SEQ ID NO: 185 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 200 (VLFW3 )). In some embodiments, the antibody molecule comprises light chain framework regions 1-3 from BAP049-hum04, BAP049-hum05, or BAP049-Clone-B (e.g., SEQ ID NO: 177 (VLFW1), SEQ ID NO: 191 (VLFW2), and SEQ ID NO: 205 (VLFW3)). In some embodiments, the antibody molecule comprises light chain framework regions 1-3 from BAP049-hum06 (e.g., SEQ ID NO: 181 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 196 (VLFW3)). In some embodiments, the antibody molecule comprises light chain framework regions 1-3 from BAP049-hum07 (e.g., SEQ ID NO: 177 (VLFW1), SEQ ID NO: 191 (VLFW2) and SEQ ID NO: 196 (VLFW3)). In some embodiments, the antibody molecule comprises light chain framework regions 1-3 from BAP049-hum08, BAP049-hum09, BAP049-hum15, BAP049-hum16, or BAP049-Clone-C (e.g., SEQ ID NO: 174 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 196 (VLFW3)). In some embodiments, the antibody molecule comprises light chain framework regions 1-3 of BAP049-hum10, BAP049-hum11, BAP049-hum14, BAP049-Clone-D, or BAP049-Clone-E (e.g., SEQ ID NO: 177 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 196 (VLFW3)). In some embodiments, the antibody molecule comprises light chain framework regions 1-3 from BAP049-hum12 (e.g., SEQ ID NO: 185 (VLFW1), SEQ ID NO: 194 (VLFW2) and SEQ ID NO: 196 (VLFW3)). In some embodiments, the antibody molecule comprises light chain framework regions 1-3 from BAP049-hum13 (e.g., SEQ ID NO: 183 (VLFW1), SEQ ID NO: 191 (VLFW2) and SEQ ID NO: 196 (VLFW3)). In some embodiments, the antibody molecule further comprises light chain framework 4 (VLFW4) of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049 -hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B BAP049-Clone-C BAP049 -Clone-D or BAP049-Clone-E (eg SEQ ID NO: 208).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum01 or BAP049-Clone-A (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2), and SEQ ID NO: 162 (VHFW3)) and light chain frameworks 1-3 from BAP049-hum01 or BAP049-Clone-A (e.g. SEQ ID NO: 177 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO : 202 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum02 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2) and SEQ ID NO: 162 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum02 (eg, SEQ ID NO: 185 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 200 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum03 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 157 (VHFW2) and SEQ ID NO: 166 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum03 (eg, SEQ ID NO: 185 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 200 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum04 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 157 (VHFW2) and SEQ ID NO: 166 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum04 (eg, SEQ ID NO: 177 (VLFW1), SEQ ID NO: 191 (VLFW2) and SEQ ID NO: 205 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum05 or BAP049-Clone-B (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2), and SEQ ID NO: 162 (VHFW3)) and light chain frameworks 1-3 from BAP049-hum05 or BAP049-Clone-B (e.g. SEQ ID NO: 177 (VLFW1), SEQ ID NO: 191 (VLFW2) and SEQ ID NO : 205 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum06 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2) and SEQ ID NO: 162 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum06 (eg, SEQ ID NO: 181 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum07 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2), and SEQ ID NO: 162 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum07 (eg, SEQ ID NO: 177 (VLFW1), SEQ ID NO: 191 (VLFW2) and SEQ ID NO: 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum08 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 157 (VHFW2) and SEQ ID NO: 166 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum08 (eg, SEQ ID NO: 174 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum09 or BAP049-Clone-C (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2), and SEQ ID NO: 162 (VHFW3)) and light chain frameworks 1-3 from BAP049-hum09 or BAP049-Clone-C (e.g. SEQ ID NO: 174 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO : 196 (VLFW3)).

In some embodiments, an anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum10 or BAP049-Clone-D (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 157 (VHFW2), and SEQ ID NO: 166 (VHFW3)) and light chain frameworks 1-3 from BAP049-hum10 or BAP049-Clone-D (e.g. SEQ ID NO: 177 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO : 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum11 or BAP049-Clone-E (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2), and SEQ ID NO: 162 (VHFW3)) and light chain frameworks 1-3 from BAP049-hum11 or BAP049-Clone-E (e.g. SEQ ID NO: 177 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO : 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum12 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2) and SEQ ID NO: 162 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum12 (eg, SEQ ID NO: 185 (VLFW1), SEQ ID NO: 194 (VLFW2) and SEQ ID NO: 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum13 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 153 (VHFW2) and SEQ ID NO: 162 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum13 (eg, SEQ ID NO: 183 (VLFW1), SEQ ID NO: 191 (VLFW2) and SEQ ID NO: 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum14 (e.g., SEQ ID NO: 151 (VHFW1), SEQ ID NO: 157 (VHFW2) and SEQ ID NO: 166 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum14 (eg, SEQ ID NO: 177 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum15 (e.g., SEQ ID NO: 151 (VHFW1), SEQ ID NO: 157 (VHFW2) and SEQ ID NO: 166 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum15 (eg, SEQ ID NO: 174 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 196 (VLFW3)).

In some embodiments, an anti-PD-1 antibody molecule comprises heavy chain framework regions 1-3 from BAP049-hum16 (e.g., SEQ ID NO: 147 (VHFW1), SEQ ID NO: 160 (VHFW2) and SEQ ID NO: 166 (VHFW3 )) and light chain framework regions 1-3 from BAP049-hum16 (eg, SEQ ID NO: 174 (VLFW1), SEQ ID NO: 187 (VLFW2) and SEQ ID NO: 196 (VLFW3)).

In some embodiments, the anti-PD-1 antibody molecule further comprises a heavy chain framework region 4 (VHFW4) of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049 -hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B -C, BAP049-Clone-D or BAP049-Clone-E (e.g. SEQ ID NO: 169) and light chain framework 4 (VLFW4) from BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049 -hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 -A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E (e.g. SEQ ID NO: 208).

In some embodiments, an anti-PD-1 antibody molecule comprises a heavy chain framework region comprising a combination of FW1, FW2, and FW3 framework regions as shown in FIG. 5 or 7. In another embodiment, the antibody molecule comprises a light chain framework region comprising a combination of FW1, FW2, and FW3 framework regions as shown in FIG. 5 or 7. In other embodiments, the antibody molecule comprises a heavy chain framework region comprising a combination of FW1, FW2, and FW3 framework regions as shown in FIG. 5 or 7 and a light chain framework comprising a combination of FW1, FW2 and FW3 frameworks as shown in FIG. 5 or 7.

In one embodiment, the heavy or light chain variable domain, or both, of an anti-PD-1 antibody molecule comprises an amino acid sequence that is substantially identical to the amino acid sequence disclosed herein, e.g., at least 80%, 85%, 90% , 92%, 95%, 97%, 98%, 99% or greater is identical to the variable region of an antibody described herein, e.g. an antibody selected from any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04 , BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10, BAP049-hum11, BAP049-hum12, BAP049-hum13, BAP049-hum14, BAP049- hum15 , BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E; or described in Table 1 or encoded by the nucleotide sequence in Table 1; or which differs by at least 1 or 5 residues, but less than 40, 30, 20 or 10 residues from the variable region of the antibody described in this application.

In one embodiment, the heavy or light chain variable region, or both, of an anti-PD-1 antibody molecule comprises an amino acid sequence encoded by a nucleic acid sequence described herein or a nucleic acid that hybridizes to a nucleic acid sequence described herein (e.g., , the nucleic acid sequence as shown in Tables 1 and 2), or its complement, for example, under conditions of low stringency, medium stringency or high stringency, or other hybridization conditions described in this application.

In another embodiment, the PD-1 antibody molecule comprises at least one, two, three, or four antigen-binding regions, e.g., variable regions, having the amino acid sequence shown in Table 1, or a sequence substantially identical thereto (e.g., sequence at least about 85%, 90%, 95%, 99%, or more identical to it, or that differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the sequences shown in Table 1). In another embodiment, an anti-PD-1 antibody molecule comprises a VH and/or VL domain encoded by a nucleic acid having the nucleotide sequence shown in Table 1 or a sequence substantially identical thereto (e.g., a sequence at least approximately identical to it). 85%, 90%, 95%, 99% or more, or which differs by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Table 1).

In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions from a heavy chain variable region having the amino acid sequence shown in Table 1, or a sequence substantially homologous thereto (e.g., the sequence identical to it by at least about 85%, 90%, 95%, 99% or more, and/or having one, two, three or more substitutions, insertions or deletions, for example, conservative substitutions). In yet another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDR regions from a light chain variable region having the amino acid sequence shown in Table 1, or a sequence substantially homologous thereto (e.g., the sequence identical to it by at least about 85%, 90%, 95%, 99% or more, and/or having one, two, three or more substitutions, insertions or deletions, for example, conservative substitutions). In yet another embodiment, the PD-1 antibody molecule comprises at least one, two, three, four, five, or six CDR regions of the heavy and light chain variable regions having the amino acid sequence shown in Table 1), or a sequence substantially homologous to it (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical to it, and/or having one, two, three or more substitutions, insertions or deletions, for example, conservative substitutions).

In one embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDRs and/or hypervariable loops from a heavy chain variable region having the amino acid sequence of an antibody described herein, e.g., an antibody selected from any from BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10, BAP049-hum11, BAP049-hum12, BAP049 -hum13, BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E, as shown in Table 1 , or a sequence substantially identical to it (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical to it, and/or having one, two, three or more substitutions, insertions, or deletions , for example, conservative substitutions). In another embodiment, an anti-PD-1 antibody molecule comprises at least one, two, or three CDRs and/or hypervariable loops from a light chain variable region having the amino acid sequence of an antibody described herein, e.g., an antibody selected from any from BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10, BAP049-hum11, BAP049-hum12, BAP049 -hum13, BAP049-hum14, BAP049-hum15, BAP049-hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E, as shown in Table 1 , or a sequence substantially identical to it (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical to it, and/or having one, two, three or more substitutions, insertions, or deletions , for example, conservative substitutions). In one embodiment, an anti-PD-1 antibody molecule includes all six CDR regions and/or hypervariable loops described herein, such as those described in Table 1.

In one embodiment, an anti-PD-1 antibody molecule has a variable region that is identical in sequence or that differs by 1, 2, 3, or 4 amino acids from the variable region described herein (e.g., the FR region disclosed herein) .

In one embodiment, the PD-1 antibody molecule is a full-length antibody or fragment thereof (eg, Fab, F(ab') ₂ , Fv, or single chain Fv fragment (scFv)). In some embodiments, the anti-PD-1 antibody molecule is a monoclonal antibody or a single specificity antibody. The anti-PD-1 antibody molecule can also be a humanized, chimeric, camel, shark or in vitro derived antibody molecule. In one embodiment, such an anti-PD-1 antibody molecule is a humanized antibody molecule. The heavy and light chains of an anti-PD-1 antibody molecule may be full-length (for example, the antibody may include at least one, and preferably two, full-length heavy chains and at least one, and preferably two, full-length light chains), or may include an antigen-binding fragment (eg, Fab, F(ab') ₂ , Fv, single chain Fv fragment, single domain antibody, diabody (dAb), bivalent antibody or bispecific antibody or fragment thereof, single domain variant or camel antibody).

In other embodiments, the anti-PD-1 antibody molecule has a heavy chain constant region (Fc) selected from, for example, IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE heavy chain constant regions; in particular selected from, for example, heavy chain constant regions of IgG1, IgG2, IgG3 and IgG4, more specifically, heavy chain constant region of IgG1 or IgG2 (eg human IgG1, IgG2 or IgG4). In one embodiment, the heavy chain constant region is from human IgG1. In another embodiment, the anti-PD-1 antibody molecule has a light chain constant region selected from, for example, light chain kappa or lambda constant regions, preferably kappa (eg, human kappa). In one embodiment, the constant region is altered, such as mutated, to alter the properties of the anti-PD-1 antibody molecule (eg, to increase or decrease one or more of the following: Fc receptor binding, antibody glycosylation, number of cysteine residues, effector cell function, or complement function). For example, a constant region contains a mutation at positions 296 (M→Y), 298 (S→T), 300 (T→E), 477 (H→K), and 478 (N→F) to alter Fc receptor binding (e.g. , mutant positions correspond to positions 132 (M→Y), 134 (S→T), 136 (T→E), 313 (H→K), and 314 (N→F) of SEQ ID NO: 212 or 214; or positions 135 (M→Y), 137 (S→T), 139 (T→E), 316 (H→K) and 317 (N→F) SEQ ID NO: 215, 216, 217 or 218). In another embodiment, an IgG4 heavy chain constant region, e.g., human IgG4, contains a mutation at position 228 according to EU numbering (e.g., S→P), e.g., as shown in Table 3. In some embodiments, anti-PD-1 antibody molecules include human IgG4 containing a mutation at position 228 according to EU numbering (eg S→P), for example, as shown in Table 3; and a light chain kappa constant region, e.g., as shown in Table 3. In another embodiment, an IgG1 heavy chain constant region, e.g., human IgG1, contains a mutation at one or more of position 297 according to EU numbering (e.g., N→A), EU code 265 (e.g. D→A), EU code 329 (e.g. P→A), EU code 234 (e.g. L→A) or EU code 235 (e.g. L→A ), eg, as shown in Table 3. In some embodiments, anti-PD-1 antibody molecules comprise human IgG1 containing a mutation at one or more of the above positions, eg, as shown in Table 3; and a light chain kappa constant region, for example, as shown in Table 3.

In one embodiment, the anti-PD-1 antibody molecule is isolated or recombinant.

In one embodiment, the anti-PD-1 antibody molecule is a humanized antibody molecule.

In one embodiment, the anti-PD-1 antibody molecule has a T cell epitope analysis risk score of less than 700, 600, 500, 400, or less.

In one embodiment, the anti-PD-1 antibody molecule is a humanized antibody molecule and has a 300-700, 400-650, 450-600 T cell epitope analysis based risk score or risk score as described herein.

In one embodiment, an anti-PD-1 antibody molecule comprises:

(a) a heavy chain variable region (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 light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33;

(b) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 1; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32;

(c) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224, the amino acid sequence of VHCDR2 SEQ ID NO: 5, and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33; or

(d) VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32.

In some embodiments, an anti-PD-1 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising the VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and

(ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32.

In other embodiments, the anti-PD-1 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising the VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 5 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and

(ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33.

In embodiments of the above antibody molecules, VHCDR1 comprises the amino acid sequence of SEQ ID NO:1. In other embodiments, the implementation of VHCDR1 includes the amino acid sequence of SEQ ID NO:4. In other embodiments, VHCDR1 comprises the amino acid sequence of SEQ ID NO: 224.

In embodiments, the above antibody molecules have a heavy chain variable region comprising at least one framework (FW) region comprising the amino acid sequence of any of SEQ ID NOs: 147, 151, 153, 157, 160, 162, 166, or 169, or the amino acid a sequence at least 90% identical to it, or containing no more than two amino acid substitutions, insertions or deletions compared to the amino acid sequence of any of SEQ ID NOs: 147, 151, 153, 157, 160, 162, 166 or 169.

In other embodiments, the above antibody molecules have a heavy chain variable region comprising at least one framework region comprising the amino acid sequence of any of SEQ ID NOs: 147, 151, 153, 157, 160, 162, 166, or 169.

In other embodiments, the above antibody molecules have a heavy chain variable region comprising at least two, three, or four framework regions comprising the amino acid sequences of any of SEQ ID NOs: 147, 151, 153, 157, 160, 162, 166, or 169.

In other embodiments, the above antibody molecules comprise the amino acid sequence of VHFW1 SEQ ID NO: 147 or 151, the amino acid sequence of VHFW2 SEQ ID NO: 153, 157, or 160, and the amino acid sequence of VHFW3 SEQ ID NO: 162 or 166, and optionally further include the amino acid sequence VHFW4 SEQ ID NO: 169.

In other embodiments, the above antibody molecules have a light chain variable region comprising at least one framework region comprising the amino acid sequence of any of SEQ ID NOs: 174, 177, 181, 183, 185, 187, 191, 194, 196, 200, 202, 205 or 208, or an amino acid sequence at least 90% identical to it, or containing no more than two amino acid substitutions, insertions or deletions compared to the amino acid sequence of any of 174, 177, 181, 183, 185, 187, 191 , 194, 196, 200, 202, 205 or 208.

In other embodiments, the above antibody molecules have a light chain variable region comprising at least one framework region comprising the amino acid sequence of any of SEQ ID NOs: 174, 177, 181, 183, 185, 187, 191, 194, 196, 200, 202, 205 or 208.

In other embodiments, the above antibody molecules have a light chain variable region comprising at least two, three, or four framework regions comprising the amino acid sequences of any of SEQ ID NOs: 174, 177, 181, 183, 185, 187, 191, 194, 196, 200, 202, 205 or 208.

In other embodiments, the above antibody molecules comprise the amino acid sequence of VLFW1 SEQ ID NO: 174, 177, 181, 183, or 185, the amino acid sequence of VLFW2 SEQ ID NO: 187, 191, or 194, and the amino acid sequence of VLFW3 SEQ ID NO: 196, 200, 202 or 205, and optionally further include the amino acid sequence of VLFW4 SEQ ID NO: 208.

In other embodiments, the above antibodies comprise a heavy chain variable domain comprising an amino acid sequence at least 85% identical to any of SEQ ID NOs: 38, 50, 82, or 86.

In other embodiments, said antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38, 50, 82, or 86.

In other embodiments, said antibody molecules comprise a light chain variable domain comprising an amino acid sequence at least 85% identical to any of SEQ ID NOs: 42, 46, 54, 58, 62, 66, 70, 74, or 78.

In other embodiments, said antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NOs: 42, 46, 54, 58, 62, 66, 70, 74, or 78.

In other embodiments, the above antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 38.

In other embodiments, said antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 40.

In other embodiments, said antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 91.

In other embodiments, the above antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 50.

In other embodiments, the above antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 or SEQ ID NO: 102.

In other embodiments, the above antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 82.

In other embodiments, said antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 84.

In other embodiments, the above antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 86.

In other embodiments, said antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 88.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the above antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 44.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 46.

In other embodiments, the above antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 48.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 54.

In other embodiments, said antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 56.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, said antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 60.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 62.

In other embodiments, the above antibodies comprise a light chain comprising the amino acid sequence of SEQ ID NO: 64.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, said antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 68.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 70.

In other embodiments, said antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 72.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 74.

In other embodiments, the above antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 76.

In other embodiments, the above antibody molecules comprise a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 78.

In other embodiments, the above antibody molecules comprise a light chain comprising the amino acid sequence of SEQ ID NO: 80.

In other embodiments, the above antibody molecules comprise 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.

In other embodiments, the above antibody molecules comprise 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: 66.

In other embodiments, the above antibody molecules comprise 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: 70.

In other embodiments, said antibody molecules comprise 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: 70.

In other embodiments, said antibody molecules comprise 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: 46.

In other embodiments, said antibody molecules comprise 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: 46.

In other embodiments, said antibody molecules comprise 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: 54.

In other embodiments, said antibody molecules comprise 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: 54.

In other embodiments, said antibody molecules comprise 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: 58.

In other embodiments, the above antibody molecules comprise 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: 62.

In other embodiments, the above antibody molecules comprise 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: 66.

In other embodiments, the above antibody molecules comprise 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.

In other embodiments, the above antibody molecules comprise 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: 78.

In other embodiments, said antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 70.

In other embodiments, said antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 82 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, said antibody molecules comprise a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO: 86 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the above antibody molecules comprise 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.

In other embodiments, the above antibody molecules comprise 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: 56.

In other embodiments, the above antibody molecules comprise 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: 68.

In other embodiments, the above antibody molecules comprise 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: 72.

In other embodiments, the above antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 102 and a light chain comprising the amino acid sequence of SEQ ID NO: 72.

In other embodiments, said antibody molecules comprise 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.

In other embodiments, the above antibody molecules comprise 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: 48.

In other embodiments, the above antibody molecules comprise 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: 48.

In other embodiments, the above antibody molecules comprise 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: 56.

In other embodiments, the above antibody molecules comprise 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: 56.

In other embodiments, the above antibodies comprise 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: 60.

In other embodiments, the above antibody molecules comprise 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: 64.

In other embodiments, the above antibody molecules comprise 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: 68.

In other embodiments, the above antibody molecules comprise 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: 68.

In other embodiments, the above antibody molecules comprise 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: 72.

In other embodiments, the above antibody molecules comprise 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: 72.

In other embodiments, said antibody molecules comprise 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.

In other embodiments, said antibody molecules comprise 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: 80.

In other embodiments, the above antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 84 and a light chain comprising the amino acid sequence of SEQ ID NO: 72.

In other embodiments, the above antibodies comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 84 and a light chain comprising the amino acid sequence of SEQ ID NO: 68.

In other embodiments, the above antibody molecules comprise a heavy chain comprising the amino acid sequence of SEQ ID NO: 88 and a light chain comprising the amino acid sequence of SEQ ID NO: 68.

In other embodiments, the above antibody molecules are selected from Fab, F(ab') ₂ , Fv, or a single chain Fv fragment (scFv).

In other embodiments, said antibody molecules comprise a heavy chain constant region selected from IgG1, IgG2, IgG3, and IgG4.

In other embodiments, said antibody molecules comprise a light chain constant region selected from the kappa or lambda light chain constant regions.

In other embodiments, said antibody molecules comprise a human IgG4 heavy chain constant region with a mutation at position 228 according to EU numbering or position 108 of SEQ ID NO: 212 or 214 and a light chain kappa constant region.

In other embodiments, the above antibody molecules comprise a human IgG4 heavy chain constant region containing a serine to proline mutation at position 228 according to EU numbering or position 108 of SEQ ID NO: 212 or 214, and a light chain kappa constant region.

In other embodiments, the above antibody molecules comprise a human IgG1 heavy chain constant region containing an asparagine to alanine mutation at EU position 297 or position 180 of SEQ ID NO: 216, and a light chain kappa constant region.

In other embodiments, the above antibody molecules comprise a human IgG1 heavy chain constant region comprising an aspartate to alanine mutation at position 265 according to EU numbering or position 148 of SEQ ID NO: 217 and a proline to alanine mutation at position 329 according to EU numbering or position 212 of SEQ ID NO: 217, and the kappa light chain constant region.

In other embodiments, the above antibody molecules comprise a human IgG1 heavy chain constant region comprising a leucine to alanine mutation at position 234 according to EU numbering or position 117 of SEQ ID NO: 218 and a leucine to alanine mutation at position 235 according to EU numbering or position 118 of SEQ ID NO: 218, and the kappa light chain constant region.

In other embodiments, the above antibody molecules are capable of binding to human PD-1 with a dissociation constant (K _D ) of less than about 0.2 nM.

In some embodiments, the above antibody molecules bind to human PD-1 with a K _D of less than about 0.2 nM, 0.15 nM, 0.1 nM, 0.05 nM, or 0.02 nM, e.g., from about 0, 13 nM to 0.03 nM, eg, about 0.077 nM to 0.088 nM, eg, about 0.083 nM, eg, as measured by the BiaCore method.

In other embodiments, the above antibody molecules bind to cynomolgus monkey PD-1 with a K _D of less than about 0.2 nM, 0.15 nM, 0.1 nM, 0.05 nM, or 0.02 nM, e.g., from about 0 .11 nM to 0.08 nM, eg approximately 0.093 nM, eg as measured by the BiaCore method.

In some embodiments, the above antibody molecules bind to human PD-1 and to cynomolgus monkey PD-1 with a similar K _D , eg, in the nM range, eg, as measured by the BiaCore method. In some embodiments, the above antibody molecules bind to a human PD-1-Ig fusion protein with a K _D of less than about 0.1 nM, 0.075 nM, 0.05 nM, 0.025 nM, or 0.01 nM, e.g., about 0.04 nM, for example, when measured by ELISA.

In some embodiments, the above antibody molecules bind to Jurkat cells that express human PD-1 (e.g., human PD-1 transfected Jurkat cells) with a K _D less than about 0.1 nM, 0.075 nM, 0.05 nM, 0.025 nM or 0.01 nM, eg, about 0.06 nM, eg, as measured by FACS analysis.

In some embodiments, the above antibody molecules bind to cynomolgus monkey T cells with a K _D less than about 1 nM, 0.75 nM, 0.5 nM, 0.25 nM, or 0.1 nM, e.g., about 0.4 nM , for example, when measured using FACS analysis.

In some embodiments, the above antibody molecules bind to cells that express cynomolgus monkey PD-1 (e.g., cells transfected with cynomolgus monkey PD-1) with a K _D of less than about 1 nM, 0.75 nM, 0.5 nM, 0 .25 nM or 0.01 nM, eg about 0.6 nM, eg as measured by FACS analysis.

In some embodiments, the above antibody molecules do not cross-react with mouse or rat PD-1. In other embodiments, the above antibodies are cross-reactive with rhesus monkey PD-1. For example, cross-reactivity can be measured using the BiaCore method or a binding assay using cells that express PD-1 (eg, 300.19 cells expressing human PD-1). In other embodiments, the above antibody molecules bind the extracellular Ig-like domain of PD-1.

In other embodiments, the above antibody molecules are capable of reducing the binding of PD-1 to PD-L1, PD-L2, or both, or to a cell that expresses PD-L1, PD-L2, or both. In some embodiments, the above antibody molecules reduce (e.g., block) the binding of PD-L1 to a cell that expresses PD-1 (e.g., 300.19 cells expressing human PD-1) with an IC _{50 of} less than about 1.5 nM, 1 nM , 0.8 nM, 0.6 nM, 0.4 nM, 0.2 nM, or 0.1 nM, e.g., about 0.79 nM to about 1.09 nm, e.g., about 0.94 nM, or about 0.78 nM or less, such as about 0.3 nM. In some embodiments, the above antibodies reduce (e.g., block) the binding of PD-L2 to a cell that expresses PD-1 (e.g., 300.19 cells expressing human PD-1) with an IC _{50 of} less than about 2 nM, 1.5 nM, 1 nM, 0.5 nM or 0.2 nM, eg about 1.05 nM to about 1.55 nM, or about 1.3 nM or less, eg about 0.9 nM.

In other embodiments, the above antibody molecules are capable of enhancing an antigen-specific T cell response.

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

In some embodiments, the above antibody molecules increase the expression of IL-2 from cells activated with staphylococcal enterotoxin B (SEB) (e.g., at a concentration of 25 μg/ml) by at least about 2, 3, 4, 5 times, e.g., about 2-3 times, e.g., about 2-2.6 times, e.g., about 2.3 times, compared to IL-2 expression when an isotype control (e.g. IgG4) is used, e.g. when measured in an assay with SEB T-cell activation or ex vivo analysis of human whole blood.

In some embodiments, the above antibody molecules increase the expression of IFN-γ from T cells stimulated with an anti-CD3 antibody (e.g., at a concentration of 0.1 μg/ml) by at least about 2, 3, 4, 5 times, e.g., about 1.2-3.4 times, eg, about 2.3 times, compared to IFN-γ expression when an isotype control (eg, IgG4) is used, eg when measuring IFN-γ activity in an assay.

In some embodiments, the above antibody molecules increase expression of IFN-γ from SEB-activated T cells (e.g., at a concentration of 3 pg/mL) by at least about 2, 3, 4, 5-fold, e.g., about 0, 5-4.5 fold, eg about 2.5 fold, compared to IFN-γ expression when an isotype control (eg, IgG4) is used, eg when measuring IFN-γ activity in an assay.

In some embodiments, the above antibody molecules increase expression of IFN-γ from CMV peptide-activated T cells by at least about 2, 3, 4, 5-fold, such as about 2-3.6-fold, such as about 2.8 times as compared to IFN-γ expression when an isotype control (eg, IgG4) is used, for example when measuring IFN-γ activity in an assay.

In some embodiments, the above antibody molecules increase the proliferation of CD8+ T cells activated by the CMV peptide by at least about 1, 2, 3, 4, 5-fold, e.g., about 1.5-fold, relative to the proliferation of CD8+ T- cells when an isotype control (eg, IgG4) is used, eg, when measuring the percentage of CD8+ T cells that have gone through at least n (eg, n=2 or 4) cell divisions.

In some embodiments, said antibody molecules have a Cmax of about 100 µg/ml to about 500 µg/ml, about 150 µg/ml to about 450 µg/ml, about 250 µg/ml to about 350 µg/ml, or about 200 μg/ml up to and about 400 μg/ml, for example, approximately 292.5 μg/ml, for example, when measured in a monkey.

In some embodiments, the above antibody molecules have T _1/2 from about 250 hours to about 650 hours, from about 300 hours to about 600 hours, from about 350 hours to about 550 hours, or from about 400 hours to about 500 hours, for example, approximately 465.5 hours, for example, when measured in a monkey.

In some embodiments, the above antibody molecules bind to PD-1 with a Kd slower than 5×10 ^-4 , 1×10 ^-4 , 5×10 ^-5 , or 1×10 ^-5 s ^-1 , e.g., approximately 2.13×10 ^-4 s ^-1 , for example when measured with the BiaCore method. In some embodiments, the above antibody molecules bind to PD-1 with Ka faster than 1×10 ⁴ , 5×10 ⁴ , 1×10 ⁵ , or 5×10 ⁵ M ^-1 s ^-1 , e.g., approximately 2.78×10 ⁵ M ^-1 s ^-1 , for example, when measured using the BiaCore method.

In some embodiments, the above anti-PD-1 antibody molecules bind to one or more residues in the C chain, CC' loop, C' chain, and FG loop of PD-1. The domain structure of PD-1 is described, for example, in Cheng et al., "Structure and Interactions of the Human Programmed Cell Death 1 Receptor", J. Biol. Chem. 2013, 288:11771-11785. As described by Cheng et al., chain C contains residues F43-M50, loop CC' contains residues S51-N54, chain C' contains residues Q55-F62, and loop FG contains residues L108-I114 (amino acid numbering is given according to Cheng et al., supra). Thus, in some embodiments, an anti-PD-1 antibody as described herein binds to at least one residue in one or more of the F43-M50, S51-N54, Q55-F62, and L108-I114 fragments in PD-1 . In some embodiments, an anti-PD-1 antibody as described herein binds to at least one residue in two, three, or all four fragments F43-M50, S51-N54, Q55-F62, and L108-I114 in PD-1 . In some embodiments, the anti-PD-1 antibody binds to a residue in PD-1 that is also part of the binding site for one or both of PD-L1 and PD-L2.

In another aspect of the invention, there is provided an isolated nucleic acid molecule encoding any of the above antibody molecules, as well as corresponding vectors and host cells.

Also provided is an isolated nucleic acid encoding an antibody heavy chain variable region or a light chain variable region, or both, of any of the above antibody molecules.

In one embodiment, the isolated nucleic acid encodes heavy chain CDR regions 1-3, wherein said nucleic acid comprises the nucleotide sequence of SEQ ID NOs: 108-112, 223, 122-126, 133-137, or 144-146.

In another embodiment, the isolated nucleic acid encodes light chain CDR regions 1-3, wherein said nucleic acid comprises the nucleotide sequence of SEQ ID NOs: 113-120, 127-132, or 138-143.

In other embodiments, said nucleic acid further comprises a nucleotide sequence encoding a heavy chain variable domain, wherein said nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 39, 51, 83, 87, 90, 95, or 101.

In other embodiments, said nucleic acid further comprises a nucleotide sequence encoding a heavy chain variable domain, wherein said nucleotide sequence comprises any of SEQ ID NOs: 39, 51, 83, 87, 90, 95, or 101.

In other embodiments, said nucleic acid further comprises a nucleotide sequence encoding a heavy chain, wherein said nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 41, 53, 85, 89, 92, 96, or 103.

In other embodiments, said nucleic acid further comprises a nucleotide sequence encoding a heavy chain, wherein said nucleotide sequence comprises any of SEQ ID NOs: 41, 53, 85, 89, 92, 96, or 103.

In other embodiments, said nucleic acid further comprises a nucleotide sequence encoding a light chain variable domain, wherein said nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 43, 47, 55, 59, 63, 67, 71, 75 , 79, 93, 97, 99, 104 or 106.

In other embodiments, said nucleic acid further comprises a nucleotide sequence encoding a light chain variable domain, wherein said nucleotide sequence comprises any of SEQ ID NOs: 43, 47, 55, 59, 63, 67, 71, 75, 79, 93, 97 , 99, 104 or 106.

In other embodiments, said nucleic acid further comprises a nucleotide sequence encoding a light chain, wherein said nucleotide sequence is at least 85% identical to any of SEQ ID NOs: 45, 49, 57, 61, 65, 69, 73, 77, 81 , 94, 98, 100, 105 or 107.

In other embodiments, said nucleic acid further comprises a nucleotide sequence encoding a light chain, wherein said nucleotide sequence comprises any of SEQ ID NOs: 45, 49, 57, 61, 65, 69, 73, 77, 81, 94, 98, 100 , 105 or 107.

In some embodiments, one or more expression vectors and host cells are provided, including the nucleic acids described above.

Also provided is a method for producing an antibody molecule or a fragment thereof, comprising culturing a host cell as described in the present application under conditions suitable for gene expression.

In one aspect of the invention, a method for producing an antibody molecule described in this application is provided. The method includes: providing a PD-1 antigen (eg, an antigen comprising at least a portion of a PD-1 epitope); obtaining an antibody molecule that specifically binds to the PD-1 polypeptide; and evaluating whether the antibody molecule specifically binds to the PD-1 polypeptide, or evaluating the effectiveness of the antibody molecule in modulating, eg inhibiting, PD-1 activity. The method may further comprise administering the antibody molecule to a subject, such as a human or non-human animal.

In another aspect of the invention, compositions, eg pharmaceutical compositions, are provided that include a pharmaceutically acceptable carrier, excipient or stabilizer and at least one of the therapeutic agents, eg anti-PD-1 antibody molecules described herein. In one embodiment, a composition, eg a pharmaceutical composition, comprises a combination of an antibody molecule and one or more agents, eg a therapeutic agent or other antibody molecule, as described herein. In one embodiment, the antibody molecule is conjugated to a label or therapeutic agent.

Additional inhibitors of PD-1 and other immune checkpoint molecules

In some embodiments, the PD-1 inhibitor is an inhibitor, e.g., an anti-PD-1 antibody molecule other than the anti-PD-1 antibody molecule of Table 1. In some embodiments, the PD-1 inhibitor comprises an anti-PD-1 antibody molecule from Table 1 and an anti-PD-1 antibody molecule other than the antibody molecule of Table 1. In other embodiments, the PD-1 inhibitor is an anti-PD-1 antibody selected from nivolumab, pembrolizumab, or pidilizumab.

In some embodiments, the anti-PD-1 antibody is nivolumab. Alternative names for nivolumab include MDX-1106, MDX-1106-04, ONO-4538, or BMS-936558. In some embodiments, the anti-PD-1 antibody is nivolumab (CAS Number: 946414-94-4). Nivolumab is a fully human IgG4 monoclonal antibody that specifically blocks PD-1. Nivolumab (clone 5C4) and other human monoclonal antibodies that specifically bind to PD-1 are disclosed in US 8,008,449 and WO2006/121168. In one embodiment, the PD-1 inhibitor is nivolumab and has the sequence disclosed herein (or a sequence substantially identical or similar to it, e.g., a sequence at least 85%, 90%, 95% or greater identical to the specified sequence ).

The amino acid sequences of the heavy and light chains of nivolumab are as follows:

heavy chain

QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 242)

light chain

EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSQTYSLSSTLTLSKADYEKHKVYACEVTHNKSS4

In some embodiments, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab (also called lambrolizumab, MK-3475, MK03475, SCH-900475 or Keytruda®; Merck) is a humanized IgG4 monoclonal antibody that binds to PD-1. Pembrolizumab and other humanized anti-PD-1 antibodies are disclosed in Hamid, O. et al. (2013) New England Journal of Medicine 369 (2): 134-44, US 8,354,509 and WO2009/114335. The amino acid sequences of the heavy and light chains of pembrolizumab are as follows:

Heavy chain (SEQ ID NO: 244)

QVQLVQSGVE VKKPGASVKV SCKASGYTFT NYYMYWVRQA PGQGLEWMGG 50 INPSNGGTNF NEKFKNRVTL TTDSSTTTAY MELKSLQFDD TAVYYCARRD one hundred YRFDMGFDYW GQGTTVTVSS ASTKGPSVFP LAPCSRSTSE STAALGCLVK 150 DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTKT 200 YTCNVDHKPS NTKVDKRVES KYGPPCPPCP APEFLGGPSV FLFPPKPKDT 250 LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREQFNSTY 300 RVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT 350 LPPSQEEMTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS 400 DGSFFLYSRL TVDKSRWQEG NVFSCSVMHE ALHNHYTQKS LSLSLGK 447

Light chain (SEQ ID NO: 245)

EIVLTQSPAT LSLSPGERAT LSCRASKGVS TSGYSYLHWY QQKPGQAPRL 50 LIYLASYLES GVPARFSGSG SGTDFTLTIS SLEPEDFAVY YCQHSRDLPL one hundred TFGGGTKVEI KRTVAAPSVF IFPSDEQLK SGTASVVCLL NNFYPREAKV 150 QWKVDNALQS GNSQESVTEQ DSKDSTYSLS STTLSKADY EKHKVYACEV 200 THQGLSSPVT KSFNRGEC 218'

In one embodiment, the PD-1 inhibitor is pembrolizumab as disclosed, for example, in US 8,354,509 and WO 2009/114335, and having a sequence disclosed in this application (or a sequence substantially identical or similar to it, for example, a sequence of at least 85%, 90%, 95% or greater identical to the specified sequence).

In some embodiments, the anti-PD-1 antibody is pidilizumab. Pidilizumab (CT-011; Cure Tech) is a humanized monoclonal IgG1k antibody that binds to PD-1. Pidilizumab and other humanized anti-PD-1 monoclonal antibodies are disclosed in WO2009/101611.

Other anti-PD-1 antibodies include AMP 514 (Amplimmune), among others, for example, anti-PD-1 antibodies disclosed in US 8,609,089, US 2010028330 and/or US 20120114649. Additional examples of anti-PD-1 antibodies also include, for example, REGN2810 , BGB-108 and BGB-A317.

In some embodiments, the PD-1 inhibitor is an immunoadhesin (e.g., an immunoadhesin that includes an extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region (e.g., the Fc region of an immunoglobulin sequence). In some embodiments, In one embodiment, the PD-1 inhibitor is AMP-224 (B7-DCIg; Amplimmune; e.g. disclosed in WO2010/027827 and WO2011/066342), a PD-L2 Fc fusion soluble receptor that blocks the interaction between PD-1 and B7-H1.

Examples of PD-L1 or PD-L2 inhibitors

In one embodiment, the combination described in this application includes a PD-L1 or PD-L2 inhibitor. In some embodiments, the implementation of the combination is used to treat cancer, such as cancer described in this application, such as a solid tumor or hemoblastosis. In some embodiments, the cancer is thyroid cancer (eg, anaplastic thyroid carcinoma), endometrial cancer, lymphoma, lung cancer (eg, non-small cell lung cancer), breast cancer (eg, triple-negative breast cancer), or MSI-H cancer. . In some embodiments, the PD-L1 inhibitor is an anti-PD-L1 antibody or a fragment thereof.

In an embodiment, the anti-PD-L1 antibody, or fragment thereof, is the anti-PD-L1 antibody molecule disclosed in US Patent Application Publication 2016/0108123 (USSN 14/881,888) entitled "Antibody Molecules to PD-L1 and Uses Thereof", in its entirety included by reference.

In one embodiment, an anti-PD-L1 antibody molecule comprises at least one or two heavy chain variable domains (optionally including a constant region), at least one or two light chain variable domains (optionally including a constant region), or both. other, comprising the amino acid sequence of 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 as described in Table 1 US 2016/0108123, or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-PD-L1 antibody molecule comprises at least one, two, or three complementarity determining regions (CDRs) from a heavy chain variable region and/or a light chain variable region of an antibody described herein, e.g. , 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; or as described in Table 1 US 2016/0108123, or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-PD-L1 antibody molecule comprises at least one, two, or three CDR regions (or all CDR regions simultaneously) from a heavy chain variable region comprising the amino acid sequence shown in Table 1 of US 2016/0108123 or encoded by the nucleotide sequence shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, for example, amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or the encoded nucleotide sequence shown in Table 1.

In yet another embodiment, an anti-PD-L1 antibody molecule comprises at least one, two, or three CDR regions (or all CDR regions simultaneously) from a light chain variable region comprising the amino acid sequence shown in Table 1 of US 2016/0108123 or encoded by the nucleotide sequence shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, for example, amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or encoded by the nucleotide sequence shown in Table 1. In some embodiments, the anti-PD-L1 antibody molecule comprises a substitution in the light chain CDR region, e.g., one or more substitutions in CDR1, CDR2, and /or light chain CDR3.

In another embodiment, an anti-PD-L1 antibody molecule comprises at least one, two, three, four, five, or six CDR regions (or all CDR regions simultaneously) from a heavy and light chain variable region comprising the amino acid sequence shown in Table 1 or encoded by the nucleotide sequence shown in Table 1 US 2016/0108123. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or the encoded nucleotide sequence shown in Table 1.

In one embodiment, an anti-PD-L1 antibody molecule comprises:

(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 amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3, each of which is disclosed in Table 1 US 2016/0108123; and

(ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 9, the amino acid sequence of VLCDR2 SEQ ID NO: 10, and the amino acid sequence of VLCDR3 SEQ ID NO: 11, each of which is disclosed in Table 1 US 2016/0108123 .

In another embodiment, the anti-PD-L1 antibody molecule comprises:

(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 amino acid sequence of VHCDR2 SEQ ID NO: 5 and the amino acid sequence of VHCDR3 SEQ ID NO: 3, each of which is disclosed in Table 1 US 2016/0108123; and

(ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14, each of which is disclosed in Table 1 US 2016/0108123 .

In one embodiment, the PD-L1 antibody molecule comprises the amino acid sequence of VHCDR1 SEQ ID NO: 1 as disclosed in Table 1 of US 2016/0108123. In another embodiment, the anti-PD-L1 antibody molecule comprises the amino acid sequence of VHCDR1 SEQ ID NO: 4 as disclosed in Table 1 of US 2016/0108123. In yet another embodiment, the PD-L1 antibody molecule comprises the amino acid sequence VHCDR1 of SEQ ID NO: 195, each of which is disclosed in Table 1 of US 2016/0108123.

В одном варианте осуществления молекула антитела против PD-L1 включает аминокислотную последовательность VH EVQLVQSGAEVKKPGATVKISCKVSGYTFTSYWMYWVRQARGQRLEWIGRIDPNSGSTKYNEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDYRKGLYAMDYWGQGTTVTVSS (SEQ ID NO: 316) или SEQ ID NO: 78 из US 2016/0108123 и аминокислотную последовательность VL AIQLTQSPSSLSASVGDRVTITCKASQDVGTAVAWYLQKPGQSPQLLIYWASTRHTGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQQYNSYPLTFGQGTKVEIK (SEQ ID NO: 317) или SEQ ID NO: 82 of US 2016/0108123, each of which is disclosed in Table 1 of US 2016/0108123. В одном варианте осуществления молекула антитела против PD-L1 включает аминокислотную последовательность VH EVQLVQSGAEVKKPGATVKISCKVSGYTFTSYWMYWVRQATGQGLEWMGRIDPNSGSTKYNEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCARDYRKGLYAMDYWGQGTTVTVSS (SEQ ID NO: 318) или SEQ ID NO: 30 из US 2016/0108123 и аминокислотную последовательность VL DVVMTQSPLSLPVTLGQPASISCKASQDVGTAVAWYQQKPGQAPRLLIYWASTRHTGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYPLTFGQGTKVEIK (SEQ ID NO: 319) или SEQ ID NO: 66 of US 2016/0108123, each of which is disclosed in Table 1 of US 2016/0108123.

In one embodiment, the anti-PD-L1 antibody molecule comprises the heavy chain amino acid sequence SEQ ID NO: 80 from US 2016/0108123 and the light chain amino acid sequence SEQ ID NO: 84 from US 2016/0108123, each of which is disclosed in US 2016 Table 1 /0108123. In one embodiment, the PD-L1 antibody molecule comprises the heavy chain amino acid sequence SEQ ID NO: 96 from US 2016/0108123 and the light chain amino acid sequence SEQ ID NO: 68 from US 2016/0108123, each of which is disclosed in US 2016 Table 1 /0108123.

The PD-L1 inhibitor (e.g., the anti-PD-L1 antibody molecule described herein) or the composition described herein may be incorporated into a dosage form (e.g., dosage formulation or dosage form) suitable for administration (e.g., intravenous administration) to the subject, as described in this application.

In some embodiments, the dosage form is a liquid dosage form. In other embodiments, the dosage form is a lyophilized dosage form, such as lyophilized or dried from a liquid dosage form. In other embodiments, the dosage form is a reconstituted dosage form, such as reconstituted from a lyophilized dosage form.

In some embodiments, the dosage form includes a PD-L1 inhibitor (eg, an anti-PD-L1 antibody molecule) and a buffering agent.

In some embodiments, the dosage form comprises a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule) present at a concentration of 10-500 mg/mL, e.g., 25-250 mg/mL, 50-150 mg/mL, 50- 100 mg/ml, or 100-200 mg/ml, such as 50 mg/ml, 100 mg/ml, 150 mg/ml or 200 mg/ml. In some embodiments, the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is present at a concentration of 50-150 mg/mL, such as 100 mg/mL. In some embodiments, the implementation of the dosage form has a pH of 5-6 (for example, 5.5). In some embodiments, the implementation of the dosage form is for intravenous infusion, for example, after dilution. In some embodiments, the dosage form includes one or more pharmaceutically acceptable excipients.

In some embodiments, the implementation of the dosage form includes a buffer substance, including histidine (for example, histidine buffer). In some embodiments, the dosage form includes L-histidine, L-histidine hydrochloride monohydrate, or both. In some embodiments, the buffering agent (e.g., histidine buffer) is present at a concentration of 1 mM to 20 mM, e.g., 2 mM to 15 mM, 3 mM to 10 mM, 4 mM to 9 mM, 5 mM to 8 mM mM or from 6 mM to 7 mM, e.g. mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM or 20 mM.

In some embodiments, the dosage form further comprises a carbohydrate. In some embodiments, the carbohydrate is sucrose. In some embodiments, the carbohydrate (e.g., sucrose) is present at a concentration of 50 mM to 150 mM, e.g., 25 mM to 150 mM, 50 mM to 100 mM, 60 mM to 90 mM, 70 mM to 80 mM, or 70 mM to 75 mM, e.g. 25 mM, 50 mM, 60 mM, 70 mM, 73.3 mM, 80 mM, 90 mM, 100 mM or 150 mM.

In some embodiments, the dosage form further comprises a surfactant. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the surfactant, or polysorbate 20, is present at a concentration of 0.005-0.025% (w/w), such as 0.0075-0.02% or 0. 01-0.015% (w/w), such as 0.005%, 0.0075%, 0.01%, 0.013%, 0.015% or 0.02% (w/w).

In some embodiments, the dosage form includes a PD-L1 inhibitor (eg, an anti-PD-L1 antibody molecule), a histidine buffer, a carbohydrate, and a surfactant. In some embodiments, the dosage form comprises a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule) and one or more (e.g., two, three, or all) of L-histidine, L-histidine hydrochloride monohydrate, sucrose, or polysorbate 20, for example, at pH 5-6. In some embodiments, the dosage form comprises a PD-L1 inhibitor (eg, an anti-PD-L1 antibody molecule), L-histidine, L-histidine hydrochloride monohydrate, sucrose, and polysorbate 20, eg, at pH 5-6.

The dosage form described in this application may be stored in a container. In some embodiments, 50 mg to 150 mg is present in a container (e.g., vial), e.g., 80 mg to 120 mg, 90 mg to 110 mg, 100 mg to 120 mg, 100 mg to 110 mg, 110 mg to 120 mg or 110 mg to 130 mg of a PD-L1 inhibitor (eg anti-PD-L1 antibody molecules).

Other examples of buffer substances that can be used in the dosage form described in this application include, without limitation, arginine buffer, citrate buffer or phosphate buffer. Other examples of carbohydrates that can be used in the dosage form described in this application include, without limitation, trehalose, mannitol, sorbitol, or a combination thereof. The dosage form described herein may also contain a tonicity agent, eg sodium chloride, and/or a stabilizing agent, eg an amino acid (eg, glycine, arginine, methionine, or a combination thereof).

In one embodiment, the combination comprises a PD-L1 inhibitor or an antibody molecule disclosed in US 2016/0108123 and a PD-1 inhibitor (eg, an anti-PD-1 antibody described herein).

The combination of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein) and a PD-L1 inhibitor (eg, an anti-PD-L1 antibody molecule described herein) was tested in the MC38 mouse colon adenocarcinoma model . The study used a surrogate antibody against mouse PD-1, RMP1-14. Co-administration of RMP1-14 and an anti-PD-L1 antibody molecule resulted in increased antitumor activity compared to single agents in this model. For example, an anti-PD-L1 and RMP1-14 antibody molecule in combination resulted in 4/10 animals with complete responses as well as 3/10 animals showing partial responses. Combination therapy resulted in a non-significant improvement in median time to end point to 46 days, which was 21 days longer than isotype-treated animals and more than 20 days longer than when one of the antibodies was administered alone.

In some embodiments, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) binds PD-1 and inhibits its interaction with PD-L1, PD-L2, or both. In some embodiments, a PD-L1 inhibitor (eg, an anti-PD-L1 antibody molecule) binds PD-L1 and inhibits its interaction with PD-1, B7.1, or both. PD-1 and PD-L1 mediate overlapping but also distinct functions on the T cell. Both receptors can transmit negative signals to the T cell upon binding. PD-L1 can mediate an immunosuppressive function when interacting with B7.1, such as blocking the ability of B7.1 to activate T cells by binding to CD28 and further stop the generation of an immune response (Butte et al. Immunity. 2007; 27(1):111 -22). Without being limited by theory, it is believed that in some embodiments, the implementation of the combined inhibition of PD-L1 and PD-1 can lead to non-redundant and/or additional antitumor immunity (Dong et al. Nat Med. 2002; 8(8):793-800; Yamazaki et al J Immunol 2002;169(10):5538-45; Taube et al Sci Transl Med 2012;4(127):127ra37). In some embodiments, the combination of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule described herein) increases T cell activation. compared to a PD-1 inhibitor or a PD-L1 inhibitor alone. In some embodiments, the combination of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein) and a PD-L1 inhibitor (eg, an anti-PD-L1 antibody molecule described herein) has synergistic antitumor activity.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose (e.g., constant dose) of 10 mg to 2000 mg, e.g., 20 mg to 1600 mg, 40 mg to 1200 mg, 80 mg to 800 mg, 100 mg to 600 mg, 200 mg to 300 mg, 20 mg to 100 mg, 40 mg to 200 mg, 100 mg to 400 mg, 200 mg up to 600 mg, 800 mg to 1200 mg, or 1200 mg to 1600 mg, e.g. at a dose (e.g. constant dose) of approximately 10 mg, 20 mg, 40 mg, 80 mg, 100 mg, 150 mg, 200 mg, 240 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, 1000 mg, 1200 mg, 1600 mg or 2000 mg, for example, once every three weeks, once every four weeks, or once every six weeks.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of 20 mg to 1200 mg (e.g., 80 mg to 800 mg, e.g., about 20 mg, 80 mg , 240 mg, 800 mg or 1200 mg), for example, once every three weeks or once every six weeks. For example, a PD-L1 inhibitor or an anti-PD-L1 antibody molecule disclosed in US 2016/0108123 may be administered at a dose of 10 mg to 100 mg (e.g., 20 mg to 80 mg, e.g., 20 mg, 40 mg, or 80 mg), for example, once every three weeks or once every six weeks. In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered by intravenous infusion.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of 10 mg to 100 mg (e.g., 20 mg to 80 mg, e.g., 20 mg, 40 mg, or 80 mg), for example, once every three weeks or once every six weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 50 mg to 500 mg (for example , from 100 mg to 400 mg, for example, at a dose of about 100 mg, 200 mg, 300 mg or 400 mg), for example, once every 3 weeks or once every 4 weeks, for example, by intravenous infusion. In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 100 mg to 300 mg (e.g., a dose of about 100 mg, 200 mg, or 300 mg), e.g., once every 3 weeks eg by intravenous infusion. In some embodiments, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of approximately 20 mg, e.g. once every three weeks, and the PD-1 inhibitor (e.g., anti-PD-L1 antibody molecule -1) is administered at a dose of approximately 100 mg or 300 mg, for example once every three weeks. In some embodiments, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of about 20 mg, 80 mg, 240 mg, 800 mg, or 1200 mg, such as once every three weeks, and a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered at a dose of about 300 mg, eg, once every three weeks.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of 10 mg to 50 mg (e.g., about 20 mg), such as once every three weeks or once at six weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 50 mg to 200 mg (for example, approximately 100 mg), for example, once every 3 weeks, for example, by intravenous infusion.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of 10 mg to 50 mg (e.g., about 20 mg), such as once every three weeks or once at six weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 400 mg (for example, approximately 300 mg), for example, once every 3 weeks, for example, by intravenous infusion.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of 50 mg to 100 mg (e.g., about 80 mg), such as once every three weeks or once at six weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 400 mg (for example, approximately 300 mg), for example, once every 3 weeks, for example, by intravenous infusion.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of 200 mg to 300 mg (e.g., about 240 mg), such as once every three weeks or once at six weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 400 mg (for example, approximately 300 mg), for example, once every 3 weeks, for example, by intravenous infusion.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of 500 mg to 1000 mg (e.g., about 800 mg), e.g., once every three weeks or once at six weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 400 mg (for example, approximately 300 mg), for example, once every 3 weeks, for example, by intravenous infusion.

In one embodiment, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered at a dose of 1000 mg to 1500 mg (e.g., about 1200 mg), e.g., once every three weeks or once at six weeks, for example, by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 400 mg (for example, approximately 300 mg), for example, once every 3 weeks, for example, by intravenous infusion.

In some embodiments, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered prior to administration of the PD-1 inhibitor (eg, anti-PD-1 antibody molecule). For example, a PD-L1 inhibitor or an anti-PD-L1 antibody molecule disclosed in US 2016/0108123 may be administered first, followed by an infusion of a dextrose solution for approximately 1 hour before the start of administration of the PD-1 inhibitor (e.g., an anti-PD-L1 antibody molecule). 1). In some embodiments, the PD-L1 inhibitor or anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered after administration of the PD-1 inhibitor (eg, anti-PD-1 antibody molecule).

In one embodiment, a PD-L1 inhibitor or an anti-PD-L1 antibody molecule disclosed in US 2016/0108123 is administered in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat a solid tumor, e.g., lung cancer. (eg, non-small cell lung cancer), breast cancer (eg, triple negative breast cancer), uterine cancer (eg, endometrial carcinoma), or thyroid cancer (eg, anaplastic thyroid carcinoma).

In some embodiments, the PD-L1 inhibitor is an antibody molecule. In some embodiments, the anti-PD-L1 inhibitor is selected from YW243.55.S70, MPDL3280A, MEDI-4736, MSB-0010718C, or MDX-1105.

In some embodiments, the anti-PD-L1 antibody is MSB0010718C. MSB0010718C (also referred to as A09-246-2; Merck Serono) is a monoclonal antibody that binds to PD-L1. Pembrolizumab and other humanized anti-PD-L1 antibodies are disclosed in WO2013/079174 and have the sequence disclosed in this application (or a sequence substantially identical or similar to it, for example, a sequence of at least 85%, 90%, 95% or above identical to the specified sequence). The heavy and light chain amino acid sequences of MSB0010718C include at least the following:

Heavy chain (SEQ ID NO: 24 as disclosed in WO2013/079174)

EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQAPGKGLEWVSSIYPSGGITFYADKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARIKLGTVTTVDYWGQGTLVTVSS (SEQ ID NO: 246)

Light chain (SEQ ID NO: 25 as disclosed in WO2013/079174)

QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIYDVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYTSSSTRVFGTGTKVTVL (SEQ ID NO: 247)

In one embodiment, the PD-L1 inhibitor is YW243.55.S70. Antibody YW243.55.S70 is an anti-PD-L1 antibody described in WO 2010/077634 (the heavy and light chain variable region sequences are shown in SEQ ID NOs. 20 and 21, respectively) and has the sequence disclosed in said application ( or a sequence substantially identical or similar to it, for example, a sequence at least 85%, 90%, 95% or higher identical to the specified sequence).

In one embodiment, the PD-L1 inhibitor is MDX-1105. MDX-1105, also known as BMS-936559, is an anti-PD-Ll antibody described in WO2007/005874 and has the sequence disclosed in said application (or a sequence substantially identical or similar to it, e.g., a sequence of at least 85%, 90%, 95% or greater identical to the specified sequence).

In one embodiment, the PD-L1 inhibitor is MDPL3280A (Genentech/Roche). MDPL3280A is a human Fc-optimized IgG1 monoclonal antibody that binds to PD-L1. MDPL3280A and other anti-PD-L1 human monoclonal antibodies are disclosed in US Patent 7,943,743, PCT Publication WO 2013/019906, and U.S. 2012/0039906. For example, MDPL3280A (also known as atezolizumab or RO5541267) may include a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 24 as disclosed in WO 2013/019906 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 21, as disclosed in WO 2013/019906 (or a sequence substantially identical or similar to it, for example, a sequence at least 85%, 90%, 95% or higher identical to the specified sequence).

In one embodiment, the PD-L1 inhibitor is MEDI-4736 (also known as durvalumab). MEDI-4736 is described in WO 2011/066389 and WO 2015/036499. For example, MEDI-4736 may include a light chain variable region comprising the amino acid sequence of SEQ ID NO: 1 as disclosed in WO 2015/036499 and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2 as disclosed in WO 2015/ 036499 (or a sequence substantially identical or similar to it, for example, a sequence at least 85%, 90%, 95% or higher identical to the specified sequence).

In other embodiments, the PD-L2 inhibitor is AMP-224. AMP-224 is a PD-L2 Fc fused soluble receptor that blocks the interaction between PD-1 and B7-H1 (B7-DCIg; Amplimmune; eg disclosed in WO2010/027827 and WO2011/066342).

Examples of LAG-3 inhibitors

In one embodiment, the combination described in this application includes a LAG-3 inhibitor. In some embodiments, the implementation of the combination is used to treat cancer, such as cancer described in this application, such as a solid tumor or hemoblastosis. In some embodiments, the cancer is lung cancer (eg, non-small cell lung cancer), skin cancer (eg, melanoma), or kidney cancer (eg, renal cell carcinoma). In some embodiments, the LAG-3 inhibitor is an anti-LAG-3 antibody or a fragment thereof.

In one embodiment, the anti-LAG-3 antibody or fragment thereof is an anti-LAG-3 antibody molecule as described in US 2015/0259420 entitled "Antibody Molecules to LAG-3 and Uses Thereof", incorporated by reference in its entirety.

In one embodiment, an anti-LAG-3 antibody molecule comprises at least one, two, three, four, five, or six CDR regions (or all CDR regions simultaneously) from the heavy and light chain variable regions (e.g., at least one , two or three CDRs from a heavy chain variable region, at least one, two or three CDRs from a light chain variable region, or both) from an antibody selected from any of BAP050-hum01, BAP050-hum02 BAP050-hum03 BAP050-hum04 BAP050-hum05 BAP050-hum06 BAP050-hum07 BAP050-hum08 BAP050-hum09 BAP050-hum10 BAP050-hum11 BAP050-hum12 BAP050-hum13 BAP050-hum14 -hum15, BAP050-hum16, BAP050-hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) (for example BAP050-hum01-Ser, BAP050-hum02-Ser, BAP050-hum03-Ser, BAP050- hum04-Ser BAP050-hum05-Ser BAP050-hum06-Ser BAP050-hum07-Ser BAP050-hum08-Ser BAP050-hum09-Ser BAP050-hum10-Ser BAP050-hum11-Ser BAP050-hum12- Ser, BAP050-hum13-Ser, BAP050- hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050-Clone-H, BAP050-Clone -I or BAP050-Clone-J; or as described in Table 1 US 2015/0259420; or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences, or closely related CDR- regions, eg CDR regions, that are identical or that have at least one amino acid change but no more than two, three or four changes (eg substitutions, deletions or insertions, eg conservative substitutions).

In another embodiment, an anti-LAG-3 antibody molecule comprises at least one, two, three, or four variable regions (e.g., at least one or two heavy chain variable regions (optionally including a constant region), at least one or two light chain variable regions (optionally including a constant region), or both) from an antibody described herein, e.g., an antibody selected from any of BAP050-hum01, BAP050-hum02, BAP050-hum03, BAP050-hum04, BAP050-hum05 BAP050-hum06 BAP050-hum07 BAP050-hum08 BAP050-hum09 BAP050-hum10 BAP050-hum11 BAP050-hum12 BAP050-hum13 BAP050-hum14 BAP050-hum15 hum17, BAP050-hum18, BAP050-hum19, BAP050-hum20, huBAP050(Ser) , BAP050-hum06-Ser, BAP050-hum07-Ser, BAP050-hum08-Ser, BAP050-hum09-Ser, BAP050-hum10-Ser, BAP050-hum11-Ser, BAP050-hum12-Ser, B AP050-hum13-Ser, BAP050-hum14-Ser, BAP050-hum15-Ser, BAP050-hum18-Ser, BAP050-hum19-Ser or BAP050-hum20-Ser), BAP050-Clone-F, BAP050-Clone-G, BAP050 -Clone-H, BAP050-Clone-I or BAP050-Clone-J; or as described in Table 1 US 2015/0259420; or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, an anti-LAG-3 antibody molecule comprises at least one, two, or three CDR regions (or all CDR regions simultaneously) from a heavy chain variable region comprising the amino acid sequence shown in Table 1 of US 2015/0259420 or encoded by the nucleotide sequence shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, for example, amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or the encoded nucleotide sequence shown in Table 1.

In yet another embodiment, an anti-LAG-3 antibody molecule comprises at least one, two, or three CDR regions (or all CDR regions simultaneously) from a light chain variable region comprising the amino acid sequence shown in Table 1 of US 2015/0259420 or encoded by the nucleotide sequence shown in Table 1. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, for example, amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or encoded by the nucleotide sequence shown in Table 1. In some embodiments, the anti-PD-L1 antibody molecule comprises a substitution in the light chain CDR region, e.g., one or more substitutions in CDR1, CDR2, and /or light chain CDR3.

In another embodiment, an anti-LAG-3 antibody molecule comprises at least one, two, three, four, five, or six CDR regions (or all CDR regions simultaneously) from a heavy and light chain variable region comprising the amino acid sequence shown in Table 1 or encoded by the nucleotide sequence shown in Table 1 US 2015/0259420. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid sequence shown in Table 1 or the encoded nucleotide sequence shown in Table 1.

In one embodiment, an anti-LAG-3 antibody molecule comprises:

(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: 286; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3, each of which is disclosed in Table 1 US 2015/0259420; and

(ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 12, each of which is disclosed in Table 1 US 2015/0259420 .

In another embodiment, the anti-LAG-3 antibody molecule comprises:

(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: 286; the amino acid sequence of VHCDR2 SEQ ID NO: 5 and the amino acid sequence of VHCDR3 SEQ ID NO: 3, each of which is disclosed in Table 1 of US 2015/0259420; and

(ii) a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 15, each of which is disclosed in Table 1 US 2015/0259420 .

In one embodiment, the anti-LAG-3 antibody molecule comprises the amino acid sequence of VHCDR1 of SEQ ID NO:1. In another embodiment, the anti-LAG-3 antibody molecule comprises the amino acid sequence of VHCDR1 of SEQ ID NO:4. In another embodiment, the anti-LAG-3 antibody molecule comprises the amino acid sequence VHCDR1 of SEQ ID NO: 286, each of which is disclosed in Table 1 of US 2015/0259420.

В некоторых вариантах осуществления молекула антитела против LAG-3 включает VH, включающую аминокислотную последовательность QVQLVQSGAEVKKPGASVKVSCKASGFTLTNYGMNWVRQARGQRLEWIGWINTDTGEPTYADDFKGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCARNPPYYYGTNNAEAMDYWGQGTTVTVSS (SEQ ID NO: 320) или SEQ ID NO: 104 из US 2015/0259420, и VL, включающую аминокислотную последовательность DIQMTQSPSSLSASVGDRVTITCSSSQDISNYLNWYLQKPGQSPQLLIYYTSTLHLGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYYNLPWTFGQGTKVEIK (SEQ ID NO: 321) or SEQ ID NO: 56 of US 2015/0259420, each of which is disclosed in Table 1 of US 2015/0259420. В некоторых вариантах осуществления молекула антитела против LAG-3 включает VH, включающую аминокислотную последовательность QVQLVQSGAEVKKPGASVKVSCKASGFTLTNYGMNWVRQAPGQGLEWMGWINTDTGEPTYADDFKGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCARNPPYYYGTNNAEAMDYWGQGTTVTVSS (SEQ ID NO: 322) или SEQ ID NO: 108 из US 2015/0259420, и VL, включающую аминокислотную последовательность DIQMTQSPSSLSASVGDRVTITCSSSQDISNYLNWYQQKPGKAPKLLIYYTSTLHLGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQQYYNLPWTFGQGTKVEIK (SEQ ID NO: 323) or SEQ ID NO: 36 of US 2015/0259420, each of which is disclosed in Table 1 of US 2015/0259420.

In some embodiments, an anti-LAG-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 122 of US 2015/0259420 and a light chain comprising the amino acid sequence of SEQ ID NO: 58 of US 2015/0259420, each of which is disclosed in Table 1 US 2015/0259420. In some embodiments, an anti-LAG-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 134 of US 2015/0259420 and a light chain comprising the amino acid sequence of SEQ ID NO: 38 of US 2015/0259420, each of which is disclosed in Table 1 US 2015/0259420.

In some embodiments, the anti-LAG-3 antibody is BMS-986016. BMS-986016 (also referred to as BMS986016; Bristol-Myers Squibb) is a monoclonal antibody that binds to LAG-3. BMS-986016 and other humanized anti-LAG-3 antibodies are disclosed in US 2011/0150892, WO2010/019570 and WO2014/008218.

Examples of TIM-3 inhibitors

In one embodiment, the combination described in this application includes a TIM-3 inhibitor. In some embodiments, the combination is used to treat cancer, such as the cancer described herein, such as a solid tumor or hemoblastosis. In some embodiments, the cancer is lung cancer (eg, non-small cell lung cancer), skin cancer (eg, melanoma), or kidney cancer (eg, renal cell carcinoma). In other embodiments, the cancer is not non-small cell lung cancer, melanoma, and renal cell carcinoma. In some embodiments, the TIM-3 inhibitor is an anti-TIM-3 antibody or fragment thereof.

In one embodiment, the anti-TIM-3 antibody, or fragment thereof, is an anti-TIM-3 antibody molecule as described in US 2015/0218274, entitled "Antibody Molecules to TIM-3 and Uses Thereof", incorporated by reference in its entirety.

In one embodiment, an anti-TIM-3 antibody molecule comprises at least one, two, three, four, five, or six CDR regions (or all CDR regions simultaneously) from a heavy and light chain variable region (e.g., at least one , two or three CDRs from a heavy chain variable region, at least one, two or three CDRs from a light chain variable region, or both) from an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3- hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4 US 2015/0218274; or encoded by the nucleotide sequence in Tables 1-4; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences, or closely related CDR- regions, eg CDR regions, that are identical or that have at least one amino acid change but no more than two, three or four changes (eg substitutions, deletions or insertions, eg conservative substitutions).

In yet another embodiment, the anti-TIM-3 antibody molecule comprises at least one, two, three, or four variable regions (e.g., at least one or two heavy chain variable regions (optionally including a constant region), at least one or two light chain variable regions (optionally including a constant region) or both) from an antibody described herein, e.g., an antibody selected from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04 , ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3 -hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4 US 2015/0218274; or encoded by the nucleotide sequence in Tables 1-4; or a sequence substantially identical (eg, at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or greater identical) to any of the above sequences.

In yet another embodiment, the anti-TIM-3 antibody molecule comprises at least one, two, or three CDR regions (or all CDR regions simultaneously) from a heavy chain variable region comprising the amino acid sequence shown in Tables 1-4 US 2015/ 0218274 or encoded by the nucleotide sequence shown in Tables 1-4. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid sequence shown in Tables 1-4 or the encoded nucleotide sequence shown in Table 1-4.

In yet another embodiment, the TIM-3 antibody molecule comprises at least one, two, or three CDR regions (or all CDR regions simultaneously) from a light chain variable region comprising the amino acid sequence shown in Tables 1-4 US 2015/ 0218274 or encoded by the nucleotide sequence shown in Tables 1-4. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid sequence shown in Tables 1-4 or the encoded nucleotide sequence shown in Tables 1-4. In some embodiments, the anti-TIM-3 antibody molecule comprises a substitution in the light chain CDR region, eg, one or more substitutions in the light chain CDR1, CDR2, and/or CDR3.

In another embodiment, an anti-TIM-3 antibody molecule comprises at least one, two, three, four, five, or six CDR regions (or all CDR regions simultaneously) from a heavy and light chain variable region comprising the amino acid sequence shown in Tables 1-4 US 2015/0218274 or encoded by the nucleotide sequence shown in Tables 1-4. In one embodiment, one or more CDR regions (or all CDR regions simultaneously) have one, two, three, four, five, six or more changes, such as amino acid substitutions or deletions, compared to the amino acid sequence shown in Tables 1-4 or the encoded nucleotide sequence shown in Tables 1-4.

In one embodiment, an anti-TIM-3 antibody molecule comprises:

(a) a heavy chain variable region (VH) comprising the amino acid sequence VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 10 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14, each of which is disclosed in Tables 1-4 US 2015/0218274 ;

(b) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 4 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8, each of which is disclosed in Tables 1-4 of US 2015/0218274;

(c) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 25 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14, each of which is disclosed in Tables 1-4 US 2015/0218274;

(d) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 24 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8, each of which is disclosed in Tables 1-4 of US 2015/0218274;

(e) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 9; the amino acid sequence of VHCDR2 SEQ ID NO: 31 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 12, the amino acid sequence of VLCDR2 SEQ ID NO: 13, and the amino acid sequence of VLCDR3 SEQ ID NO: 14, each of which is disclosed in Tables 1-4 US 2015/0218274; or

(f) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 3; the amino acid sequence of VHCDR2 SEQ ID NO: 30 and the amino acid sequence of VHCDR3 SEQ ID NO: 5; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 6, the amino acid sequence of VLCDR2 SEQ ID NO: 7, and the amino acid sequence of VLCDR3 SEQ ID NO: 8, each of which is disclosed in Tables 1-4 of US 2015/0218274.

В некоторых вариантах осуществления молекула антитела против TIM-3 включает VH, включающую аминокислотную последовательность QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGQGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS (SEQ ID NO: 324) или SEQ ID NO: 32 из US 2015/0218274, и VL, включающую аминокислотную последовательность DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIK (SEQ ID NO: 325) or SEQ ID NO: 20 of US 2015/0218274, each of which is disclosed in Table 1 of US 2015/0218274. В некоторых вариантах осуществления молекула антитела против TIM-3 включает VH, включающую аминокислотную последовательность QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS (SEQ ID NO: 326) или SEQ ID NO: 52 из US 2015/0218274, и VL, включающую аминокислотную последовательность AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIK (SEQ ID NO: 327) or SEQ ID NO: 64 of US 2015/0218274, each of which is disclosed in Table 1 of US 2015/0218274.

In some embodiments, an anti-TIM-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 34 of US 2015/0218274 and a light chain comprising the amino acid sequence of SEQ ID NO: 22 of US 2015/0218274, each of which is disclosed in Table 1 US 2015/0218274. In some embodiments, an anti-TIM-3 antibody molecule comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 54 of US 2015/0218274 and a light chain comprising the amino acid sequence of SEQ ID NO: 66 of US 2015/0218274, each of which is disclosed in Table 1 US 2015/0218274.

Examples of anti-TIM-3 antibodies are also disclosed in US Patent 8,552,156, WO 2011/155607, EP 2581113 and U.S. 2014/044728.

In one embodiment, the combination described herein comprises a TIM-3 inhibitor (eg, an anti-TIM-3 antibody molecule described herein) and a chemotherapeutic agent (eg, a hypomethylating agent, eg, decitabine). In some embodiments, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) is administered at a dose of 10 mg to 1000 mg (e.g., 20 mg to 800 mg, 80 mg to 400 mg, eg at a dose of 240 mg), eg once every 2 weeks, eg intravenously. In some embodiments, the TIM-3 inhibitor (eg, the anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) is stored in the form of a liquid (eg, vial) for intravenous infusion. In some embodiments, the chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered at a dose of 5 mg/m ² to 50 mg/m ² (e.g., 10 mg/m ² to 30 mg/m ² (e.g., in dose of 20 mg/m ² ), for example, daily, for example, for 1, 2, 3, 4, 5, 6, 7, 8 or more days, for example, intravenously. For example, a TIM-3 inhibitor (for example, an antibody molecule against TIM-3 disclosed in U.S. Patent Application Publication 2015/0218274) may be administered on Days 8 and 22 of a 28 day course, and a chemotherapeutic agent (eg, a hypomethylating agent such as decitabine) may be administered on Days 1, 2, 3 , 4, and 5 of a 28 day course In some embodiments, the chemotherapeutic agent (eg, a hypomethylating agent, eg, decitabine) is administered by intravenous infusion over 30 minutes to 2 hours, eg, 1 hour.

In one embodiment, the combination further comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein). In some embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 600 mg (e.g., 300 mg to 500 mg, e.g., 400 mg), e.g., once every 4 weeks, for example, intravenously. For example, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein) may be administered on Day 8 of a 28 day course. In some embodiments, the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is stored in the form of a lyophilisate (eg, in a vial) for intravenous infusion. In some embodiments, when administered in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein), a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in Application Publication No. US Pat. No. 2015/0218274) is administered at a dose of 20 mg to 400 mg (e.g. 40 mg to 200 mg or 50 mg to 100 mg, e.g. 80 mg), e.g. once every 2 weeks, e.g. intravenously.

In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) are administered one day. In some embodiments, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) is administered prior to administration of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein). ). In some embodiments, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) is administered first, followed by an infusion of saline or dextrose solution, e.g., over about 1 hour, up to start of administration of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein). In other embodiments, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) is administered after administration of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein). ). In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) are administered on different days.

In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein), a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274), or both are administered by intravenous infusion over 30 minutes to 2 hours, eg about 1 hour.

In some embodiments, the combination is used to treat cancer, eg, the cancer described herein, eg, hemoblastosis, eg, leukemia (eg, acute myeloid leukemia (AML)), or myelodysplastic syndrome (MDS). In some embodiments, the cancer is leukemia, eg, relapsed or refractory AML, or de novo AML (eg, substandard AML). In other embodiments, the cancer is MDS, eg high risk MDS.

Examples of CTLA-4 inhibitors

In one embodiment, the combination described in this application includes a CTLA-4 inhibitor. In some embodiments, the combination is used to treat cancer, such as the cancer described herein, such as a solid tumor or hemoblastosis.

Examples of anti-CTLA-4 antibodies include tremelimumab (IgG2 monoclonal antibody manufactured by Pfizer, formerly known as ticilimumab, CP-675,206); and ipilimumab (an anti-CTLA-4 antibody also known as MDX-010, CAS 477202-00-9).

In one embodiment, the combination comprises an anti-PD-1 antibody molecule, eg, as described herein, and an anti-CTLA-4 antibody, eg, ipilimumab. Exemplary doses that may be used include a dose of an anti-PD-1 antibody molecule of about 1-10 mg/kg, such as 3 mg/kg, and a dose of an anti-CTLA-4 antibody, such as ipilimumab, of about 3 mg/kg.

Other examples of anti-CTLA-4 antibodies are disclosed in, for example, US Pat. No. 5,811,097.

Examples of IAP inhibitors

In one embodiment, the combination described herein comprises an apoptosis inhibitor protein (IAP) inhibitor. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., solid tumor (e.g., colon and rectal cancer (CRC), lung cancer (e.g., non-small cell lung cancer (NSCLC)), breast cancer cancer (eg, triple-negative breast cancer (TNBC)), ovarian cancer, or pancreatic cancer), such as hemoblastosis (eg, multiple myeloma).

In some embodiments, the IAP inhibitor is (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl )-2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Patent 8,552,003.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl )-2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Pat. . The CIAP1 and CIAP2 proteins are components of the protein complexes of the TNF family of death receptors. Without being limited by theory, it is believed that in some embodiments, the binding of the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl )pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compounds A21), or the compound disclosed in US Pat. activate NF-κB signaling after receptor activation (Gyrd-Hansen M, Meier P (2010) Nat. Rev. Cancer p. 561-74). In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is used to enhance antitumor immunity in a subject. For example, an IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2- oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, can increase the proliferation and function of human and murine T cells in vitro after costimulation, and/or enhance response to prophylactic and therapeutic anticancer vaccines in vivo , for example, as a result of NF-κB activation (Dougan et al. (2010) J. Exp. Med. p. 2195-206). NF-κB activity has been implicated in T cell cross-priming during immunogenic apoptosis in response to the release of threat-associated inflammatory molecules (Yatim et al. (2015) Science p. 328-34), suggesting a mechanism for increased T cell function and antitumor immunity. As another example, an IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl) -2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, can protect monocytic-derived dendritic cells from apoptosis, for example, similar to CD40 ligation (Knights et al. (2013) Cancer Immunol Immunother, pp. 321-35).

In some embodiments, the combination described herein includes an IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl )pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or a compound disclosed in US Pat. No. 8,552,003, and an immune checkpoint molecule inhibitor, e.g. antibodies against PD-1).

Without being limited by theory, it is believed that in some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl) pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is an orally available small molecule that activates NF-kB signaling upon activation of members of the TNF family receptors. NF-κB is a major transcriptional regulator in immune cells and also functions directly in tumor cells (Perkins (2012) Nat. Rev. Cancer p. 121-32). Preclinical data suggest that NF-κB activity is required for cross-priming of CD8+ T lymphocytes undergoing immunogenic apoptosis (Yatim et al. (2015) Science p. 328-34). Compound A21 stimulated T-lymphocyte proliferation, induced IFN-gamma, and suppressed IL-10 production in vitro . Clinical studies with Compound A21 have demonstrated the induction of circulating cytokines including TNF alpha, IL-8, IL-10 and CCL2 (Infante et al. (2014) J. Clin. Oncol. p. 3103-10). Clinical data indicate that Compound A21 increased the complete response rate of TNBC to paclitaxel treatment according to laboratory studies. In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. antibodies against PD-1).

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or a compound disclosed in US Pat. No. 8,552,003, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), are administered at a dose and/or time schedule such that, in combination, they provide the desired antitumor activity.

Without being limited by theory, doses of the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1 -yl)-2-oxoethyl)-2-(methylamino)propanamide (Compounds A21), or the compound disclosed in US Pat. For example, paired skin biopsies from two patients receiving a 320 mg dose showed degradation of CIAP1, the target protein of Compound A21; skin biopsies from patients treated with lower doses (160 mg and below) showed less stable pharmacodynamic activity (Infante et al. (2014) J. Clin. Oncol. p. 3103-10). In some embodiments, doses above 900 mg are not required for clinical efficacy. In some situations, the 1800 mg dose of Compound A21 was poorly tolerated when administered in combination with paclitaxel. Thus, in one embodiment, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1- yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of 300 mg to 900 mg, for example, once a week. In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered less frequently than once a week, e.g., administered once every two weeks, once every three weeks, or once every four weeks.

In some embodiments, an IAP inhibitor, e.g., (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1 -yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Pat. No. 8,552,003 is administered at a dose of about 1800 mg or less, for example once a week.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Patent 8,552,003, is administered at a dose of 160 mg to 1800 mg, 200 mg to 1200 mg, 400 mg to 800 mg or 500 mg to 700 mg, such as 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, or 900 mg, such as once a week, once every other week , once every three weeks or once every four weeks.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of about 200 mg to about 400 mg, such as about 300 mg, once a week. In other embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of about 800 mg to about 1000 mg, such as about 900 mg, once a week. In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of about 200 mg to about 400 mg, such as about 300 mg, once every four weeks.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered orally.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or a compound disclosed in US Pat. No. 8,552,003, is used in combination with a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule). In one embodiment, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of 200 mg to 400 mg (e.g. at a dose of 300 mg), e.g. once a week, e.g., orally, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg), e.g., once every 4 weeks, e.g., by intravenous infusion .

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or a compound disclosed in US Pat. No. 8,552,003, is used in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat colon and rectal cancer , lung cancer (eg, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple-negative breast cancer (eg, TNBC)).

Examples of EGFR inhibitors

In one embodiment, the combination described herein comprises an epidermal growth factor receptor (EGFR) inhibitor. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., lung cancer (e.g., non-small cell lung cancer), pancreatic cancer, breast cancer (e.g., triple negative breast cancer). gland (TNBC)) or colon cancer). In some embodiments, the cancer is selected from colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS CRC)), lung cancer (e.g., non-small cell lung cancer), or breast cancer (e.g., triple negative lung cancer (TNBC). )).

In some embodiments, the EGFR inhibitor is (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)aze-pan-3-yl)-1H-benzo[ d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757.

In some embodiments, an EGFR inhibitor, e.g., (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)aze-pan-3-yl)-1H- benzo[d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is administered at a dose of 150-250 mg, for example, per day. In some embodiments, an EGFR inhibitor, such as (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[ d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40) or the compound disclosed in PCT publication WO 2013/184757 is administered at a dose of about 150, 200 or 250 mg, or about 150-200 or 200-250 mg .

In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is a covalent, irreversible tyrosine kinase inhibitor. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, inhibits EGFR activating mutations (L858R, ex19del). In other embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, does not or substantially does not inhibit wild-type (wt) EGFR. Compound A40 has shown efficacy in patients with EGFR mutant NSCLC. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, also inhibits one or more kinases from the TEC kinase family. The Tec family kinases include, for example, ITK, BMX, TEC, RLK, and BTK, and are central to T-cell receptor proliferation and chemokine receptor signaling (Schwartzberg et al. (2005) Nat. Rev. Immunol. p. 284-95) . For example, Compound A40 can inhibit ITK with a biochemical IC _{50 of} 1.3 nM. ITK is an essential enzyme for the survival of Th2 cells, and its inhibition results in a shift in the balance between Th2 and Th1 cells. Combination in vivo treatment with the ITK inhibitor ibrutinib or Compound A40 and anti-PD-L1 antibody results in superior efficacy compared to either agent alone in several models.

In some embodiments, the combination described herein includes an EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepane-3-yl )-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g. , an anti-PD-1 antibody molecule).

The combination of ITK inhibition (ibrutinib) and checkpoint inhibition is more effective than either agent alone in various syngeneic mouse models, such as those expressing ITK but not expressing BTK. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, inhibits ITK. The synergistic effect of ITK inhibition and checkpoint blockade was investigated in mouse allografts using mouse cancer cell lines (A20, CT26 and 4T1) (Sagiv-Barfi et al. (2015) Blood, pp. 2079-86). The combination of an anti-PD-L1 antibody and ibrutinib (an ITK inhibitor) was shown to be significantly more effective than either agent alone in all three models. In these experiments, the treatment effect was sustained despite administration of ibrutinib for only 8 days and administration of a total of 5 doses of anti-PD-L1 antibody. Approximately half of the mice with CT26 tumors treated with this combination were completely cured (none of the mice treated with any single agent were cured). Repeated administration of CT26 tumor inoculum to these mice demonstrated long-term antitumor memory specific to this cell line (Sagiv-Barfi et al. (2015) Blood, pp. 2079-86). In addition, tumor-specific T cells were detected in the blood and spleen of mice treated with ibrutinib and anti-PD-L1 antibody. A similar experiment was performed using Compound A40 in an A20 lymphoma model (see, for example, Example 4). The combination of Compound A40 and anti-PD-L1 antibody or ibrutinib and anti-PD-L1 antibody was more effective than single agent. Compound A40 and ibrutinib were administered for only ten days and a total of 5 doses of anti-PD-L1 antibody were administered. Compound A40 and ibrutinib were administered only briefly, with Compound A40 plus anti-PD-L1 and ibrutinib plus anti-PD-L1 on survival longer than 60 days. The combination of anti-PD-L1 antibody and Compound A40 resulted in tumor regression in A20 lymphoma allograft-bearing mice. Thus, in some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo [d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, enhances or is used to enhance the antitumor effect of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule ).

In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT Publication WO 2013/184757, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) , administered at a dose and/or time schedule that, in combination, provide the desired antitumor activity.

In one embodiment, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40) or the compound disclosed in PCT publication WO 2013/184757 is administered at a dose of 5 mg to 100 mg, e.g. 10 mg to 75 mg, 15 mg to 50 mg, 20 mg to 30 mg, 10 mg to 40 mg, 10 mg to 25 mg, or 25 mg to 40 mg, e.g. at 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30mg, 35mg, 40mg, 45mg, 50mg, 60mg, 70mg, 80mg, 90mg or 100mg e.g. twice a day, once a day, once every other day, once three days or once a week.

In one embodiment, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is administered at a dose of 10 mg to 50 mg (eg 25 mg), for example once a day. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is administered orally. In one embodiment, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT Publication WO 2013/184757, is administered at a dose of 10 mg to 50 mg (e.g. 25 mg), for example, once a day, e.g., orally, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg), e.g., once every 4 weeks, e.g., by intravenous infusion . In some embodiments, the combination is administered in one or more courses of use, eg, one or more 28 day courses of use. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is administered on day 1 to day 10 of the first course of application. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is not administered on day 11 to day 28 of the first course of application, or is not administered in any of the subsequent courses of application.

In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, is used in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat colon cancer and rectal (CRC) (eg, MSS-CRC), lung cancer (eg, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple negative breast cancer (TNBC)).

In some embodiments, the EGFR inhibitor is selected from one of more of erlotinib, gefitinib, cetuximab, panitumumab, netcitumumab, PF-00299804, nimotuzumab, or RO5083945.

Examples of mTOR inhibitors

In one embodiment, the combination described in this application includes an inhibitor of the target of rapamycin (mTOR). In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., prostate cancer, breast cancer, brain cancer, bladder cancer, pancreatic cancer, kidney cancer, or cancer liver, lung cancer (eg, small cell lung cancer or non-small cell lung cancer), respiratory/breast cancer, sarcoma, bone cancer, non-small cell lung cancer, endocrine cancer, astrocytoma, cervical cancer, neurological cancer, gastric cancer, or melanoma), e.g. , hemoblastosis (eg leukemia (eg lymphocytic leukemia), eg lymphoma, or eg multiple myeloma). In some embodiments, the cancer is selected from colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS CRC)), lung cancer (e.g., non-small cell lung cancer), or breast cancer (e.g., triple negative lung cancer (TNBC). )).

In some embodiments, the mTOR inhibitor is dactolisib (Compound A4) or 8-(6-methoxy-pyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-1 ,3-dihydro-imidazo[4,5-c]quinolin-2-one (Compound A41), or the compound disclosed in PCT Publication WO 2006/122806.

In some embodiments, the mTOR inhibitor is everolimus (also known as Afinitor®; Compound A36) or a compound disclosed in PCT publication WO 2014/085318.

In some embodiments, an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, inhibits the mammalian target of rapamycin (mTOR), serine threonine kinase, following the PI3K/AKT pathway. The mTOR pathway is deregulated in several human cancers. Without being limited by theory, it is believed that in some embodiments, the mTOR inhibitor, everolimus (Compound A36), or the compound disclosed in PCT publication WO 2014/085318, binds to an intracellular protein, FKBP-12, resulting in the formation of an inhibitory complex with the mTOR complex. 1 (mTORC1) and thus inhibition of mTOR kinase activity. Everolimus (Compound A36) may decrease the activity of ribosomal protein kinase S6 (S6K1) and/or eukaryotic initiation factor 4E binding protein (4E-BP1), downstream mTOR effectors involved in protein synthesis. S6K1 is a substrate of mTORC1 and phosphorylates activation domain 1 of the estrogen receptor, resulting in ligand-independent activation of the receptor. In addition, everolimus (Compound A36) can inhibit the expression of hypoxia-inducible factor (eg, HIF-1) and/or reduce the expression of vascular endothelial growth factor (VEGF). Inhibition of mTOR by everolimus (Compound A36) has been shown to reduce cell proliferation, angiogenesis, and glucose uptake in in vitro and/or in vivo studies . Constitutive activation of the PI3K/Akt/mTOR pathway may contribute to endocrine resistance in breast cancer. In vitro studies show that estrogen-dependent and HER2+ breast cancer cells are sensitive to the inhibitory effects of everolimus (Compounds A36), and that combined treatment with everolimus and Akt, HER2 or aromatase inhibitors increases the antitumor activity of everolimus (Compounds A36) in a synergistic manner. Two regulators of mTORC1 signaling are oncogene suppressors, tuberin sclerosis complexes 1 and 2 (TSC1, TSC2). The loss or deactivation of TSC1 or TSC2 results in the activation of downstream signaling. In tuberous sclerosis (TSC), a genetic disorder, inactivating mutations in the TSC1 or TSC2 gene lead to the formation of hamartomas throughout the body.

In some embodiments, the combination described herein includes an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., a molecule antibodies against PD-1).

Everolimus (Compound A36) has been used clinically as a direct anticancer agent at a dose of 10 mg daily to suppress mTOR activity in tumor cells, and as an immunosuppressant in patients requiring solid organ transplantation at a dose of 1.5-2.0 mg daily to suppress the function of T-lymphocytes. Without being limited by theory, it is believed that in some embodiments, at such dosages and schedules, everolimus (Compound A36) is expected to attenuate an effective antitumor response. Immunoaging is a decline in immune function that occurs in the elderly and includes a reduced response to vaccination, including influenza vaccination. The decline in immune function with age includes an increase in PD-1 positive "depleted" T lymphocytes, which show a reduced response to antigen challenge (Lages et al. (2010) Aging Cell 9, 785-798). Clinical data indicate that a dose of 5 mg per week may be immunostimulatory, reducing the percentage of PD-1-positive CD4+ and CD8+ lymphocytes compared with placebo treatment and enhancing influenza vaccine response in elderly subjects (Mannick et al. (2014) Sci. Transl. Med. Vol. 6, Issue 268, pp. 268-179). In some embodiments, an immunostimulatory dose of an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, increases or is used to increase the antitumor activity of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule).

In some embodiments, an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), are administered. at a dose and/or time schedule that, in combination, provides the desired antitumor activity.

In some embodiments, the dose of everolimus (Compound A36) approved for adult cancer indications (e.g., breast cancer (e.g., TNBC), renal cell carcinoma, and neuroendocrine tumors (e.g., atypical pulmonary carcinoid tumor) is 10 mg daily. Without limited by theory, in some embodiments, modeling and simulation based on mTOR-mediated phosphorylation of its downstream S6 kinase (S6K) is believed to indicate that a 20 mg once weekly regimen inhibits mTOR-mediated S6K phosphorylation almost completely, a regimen with administration of 5 mg once a week inhibits S6K phosphorylation by more than 50% and a regimen of 0.5 mg once a day inhibits S6K phosphorylation by approximately 38% between doses (Mannick et al. (2014) Sci. Transl. Med Vol.6, Issue 268, pp. 268ra179 Tanaka (2008) J. Clin. Oncol. p. 1596-602. Thus, in some an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 5 mg once a week (QW), for example in combination with a PD-1 inhibitor (for example, an anti-PD antibody molecule -1). In other embodiments, a dose of 0.5 mg once a day (QD) may also be used, for example, if a dose of 5 mg once a week (QW) is not well tolerated. In some embodiments, an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 5 mg or less once a week.

In some embodiments, an mTOR inhibitor such as everolimus (Compound A36) or a compound disclosed in PCT publication WO 2014/085318 is administered at a dose of about 2.5-20 mg/day. In one embodiment, a TOR inhibitor such as everolimus (Compound A36) or a compound disclosed in PCT publication WO 2014/085318 is administered at a dose of about 2.5, 5, 10, or 20 mg/day, such as about 2.5 -5, 5-10 or 10-20 mg/day.

In one embodiment, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 1 mg to 10 mg, 2 mg to 8 mg, 3 mg to 7 mg, or 4 mg to 6 mg, such as at a dose of 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or 10 mg, such as once a week, once a two weeks, once every three weeks, or once every four weeks. In one embodiment, the mTOR inhibitor, everolimus (Compound A36), or the compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 4 mg to 6 mg, eg 5 mg, eg once a week.

In another embodiment, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 0.1 mg to 1 mg, 0.2 mg to 0.8 mg, 0 .3 mg to 0.7 mg, or 0.4 mg to 0.6 mg, e.g. 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0, 6 mg, 0.7 mg, 0.8 mg, 0.9 mg or 1 mg, for example, once a day or once a week.

In some embodiments, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered orally.

In some embodiments, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 2 mg to 8 mg (e.g., 5 mg), e.g., once a week, e.g., orally, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg dose), e.g., once every 4 weeks, e.g., by intravenous infusion .

In some embodiments, an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is used in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat colon and rectal cancer. , lung cancer (eg, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple-negative breast cancer (TNBC)).

In some embodiments, the mTOR inhibitor is selected from one or more of rapamycin, temsirolimus (Torisel®), AZD8055, BEZ235, BGT226, XL765, PF-4691502, GDC0980, SF1126, OSI-027, GSK1059615, KU-0063794, WYE-354, Palomid 529 (P529), PF-04691502 or PKI-587, ridaforolimus (officially known as deferolimus, (1R,2R,4S)-4-[(2R)-2[(1R,9S,12S,15R,16E,18R ,19R,21R,23S,24E,26E,28Z,30S,32S,35R)-1,18-dihydroxy-19,30-dimethoxy-15,17,21,23,29,35-hexamethyl-2,3, 10,14,20-pentaoxo-11,36-dioxa-4-azatricyclo[30.3.1.0 ^4,9 ]hexatriaconta-16,24,26,28-tetraen-12-yl]propyl]-2-methoxycyclohexyl-dimethylphosphinate, also known as AP23573 and MK8669, and described in PCT publication WO 03/064383); everolimus (Afinitora® or RAD001); rapamycin (AY22989, Sirolimus®); simapimoda (CAS No.: 164301-51-3); (5-{2,4-bis[(3S)-3-methylmorpholin-4-yl]pyrido[2,3-d]pyrimidin-7-yl}-2-methoxyphenyl)methanol (AZD8055); 2-amino-8-[trans-4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxy-3-pyridinyl)-4-methyl-pyrido[2,3-d]pyrimidine-7(8H)- she (PF04691502, CAS Number: 1013101-36-4); N2-[1,4-dioxo-4-[[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholinium-4-yl]methoxy]butyl]-L-arginylglycyl-L -α-aspartyl-L-serine (SEQ ID NO: 237) internal salt (SF1126, CAS No.: 936487-67-1) or XL765 (SAR245409).

Other examples of mTOR inhibitors include, but are not limited to, temsirolimus; ridaforolimus (1R,2R,4S)-4-[(2R)-2[(1R,9S,12S,15R,16E,18R,19R,21R,23S,24E,26E,28Z,30S,32S,35R)- 1,18-dihydroxy-19,30-dimethoxy-15,17,21,23,29,35-hexamethyl-2,3,10,14,20-pentaoxo-11,36-dioxa-4-azatricyclo[30.3. 1.0 ^4,9 ]hexatriaconta-16,24,26,28-tetraen-12-yl]propyl]-2-methoxycyclohexyl dimethylphosphinate, also known as AP23573 and MK8669; everolimus (RAD001); rapamycin (AY22989); simapimod; (5-{2,4-bis[(3S)-3-methylmorpholin-4-yl]pyrido[2,3-d]pyrimidin-7-yl}-2-methoxyphenyl)methanol (AZD8055); 2-amino-8-[trans-4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxy-3-pyridinyl)-4-methyl-pyrido[2,3-d]pyrimidine-7(8H)- he (PF04691502); and N ² -[1,4-dioxo-4-[[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholinium-4-yl]methoxy]butyl]-L-arginylglycyl -L-α-aspartyl-L-serine (SEQ ID NO: 237) internal salt (SF1126); and XL765.

Examples of IL-15 agonists

In one embodiment, the combination described in this application includes an interleukin-15 (IL-15) agonist. In some embodiments, the combination is used to treat cancer, e.g. the cancer described herein, e.g. solid tumor (e.g. refractory solid tumor), (e.g. melanoma (e.g. metastatic or advanced melanoma), kidney cancer (e.g. renal β-cell cancer), non-small cell lung cancer, squamous cell head and neck cancer, or bladder cancer (eg, non-muscle-invasive bladder cancer)), eg, hemoblastosis (eg, leukemia, eg, acute myelogenous leukemia (eg, refractory or relapsing acute myelogenous leukemia), for example, lymphoma, for example, non-Hodgkin's lymphoma (for example, relapsed/refractory slowly progressive B-cell non-Hodgkin's lymphoma), for example, or multiple myeloma (for example, relapsed or refractory multiple myeloma)). In some embodiments, the cancer is a solid tumor.

IL-15, secreted by mononuclear phagocytes (and some other cell types) after viral infection, regulates the activation and proliferation of T and NK cells. This cytokine induces the activation of the transcriptional activators STAT3, STAT5, and STAT6 via JAK kinase signaling pathways in mast cells, T cells, and dendritic epidermal T cells. IL-15 and interleukin-2 (IL-2) are structurally similar and share many biological activities; both can bind to the same erythropoietin receptor subunits, negatively regulating each other's activity. The number of CD8+ memory T cells can be regulated by the balance between IL-15 and IL-2.

In some embodiments, the IL-15 agonist is recombinant human IL-15 (rhIL-15), such as CYP0150 (Cytune). CYP0150 is a recombinant protein composed of human IL-15 coupled to the Sushi+ domain of the human alpha chain receptor (transpresentation).

CYP0150 is disclosed, for example, in PCT publication WO 2007/046006. CYP0150 имеет следующую аминокислотную последовательность: MAPRRARGCRTLGLPALLLLLLLRPPATRGDYKDDDDKIEGRITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSGGSGGGGSGGGSGGGGSLQNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS (SEQ ID NO: 248) (раскрытую как SEQ ID NO: 60 в WO 2007/046006) или MDSKGSSQKAGSRLLLLLVVSNLLLCQGVVSTTRDYKDDDDKIEGRNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSSGGGSGGGGSGGGGSGGGGSGGGSLQITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPP (SEQ ID NO: 249) (раскрытую как SEQ ID NO: 62 в WO 2007/046006 ).

In some embodiments, the IL-15 agonist is ALT-803 (Altor BioScience). ALT-803 is a soluble IL-15N72D:IL-15RαSu/Fc complex derived from a highly productive recombinant mammalian cell line that coexpresses IL-15N72D and the IL-15RαSu/Fc fusion protein. Mutant IL-15 (N72D) has increased IL-15 biological activity (Zhu et al. 2009, J Immunol. 183:3598). Mutant IL-15N72D and a soluble IL-15Rα domain can form stable heterodimeric complexes in solution, and the complex exhibits increased biological activity (approximately 25 times more active) than uncomplexed IL-15. ALT-803 is disclosed, for example, in PCT publication WO 2012/040323 and US Pat. No. 8,507,222.

In some embodiments, the IL-15 agonist is hetIL-15 (Admune). HetIL-15 is heterodimeric human IL-15 (IL-15/sIL-15Ra). HetIL-15 is disclosed, for example, in PCT publication WO 2009/002562 and WO 2014/066527.

Examples of CD40 agonists

In one embodiment, the combination includes a CD40 agonist. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., lung cancer, esophageal cancer, melanoma, or renal cell carcinoma), e.g., hemoblastosis (e.g., leukemia (e.g., chronic lymphocytic leukemia (CLL)), for example, lymphoma (eg, non-Hodgkin's lymphoma), or multiple myeloma).

In one embodiment, the CD40 agonist is ADC-1013 (Alligator/BioInvent). ADC-1013 is a fully human IgG anti-human CD40 monoclonal antibody. CD40, an integral membrane protein present on the surface of B lymphocytes, is a member of the tumor necrosis factor receptor superfamily and is highly expressed in many cancers such as B cell malignancies. CD40 agonists, such as anti-CD40 antibodies, are able to effectively replace the activity of helper T cells (Ridge, J. et al. (1998) Nature 393: 474-478).

ADC-1013 is disclosed, for example, in PCT publication WO 2015/091853. ADC-1013 clones include, for example, 1136/1137, 1132/1133, 1148/1149, 1140/1135, 1134/1135, 1107/1108, 1142/1135, 1146/1147 and 1150/1151.

The heavy chain variable region 1132/1133 has the following amino acid sequence: EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSGIGSYGGGTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARYVNFGMDYWGQGTLVTVSS (SEQ ID NO: 250) (disclosed as SEQ ID NO: 2815/0WO: 2815 in 0915/0WO: 2815). The 1132/1133 light chain variable region has the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGTDFTLTISSLQPEDFATYYCQQYGRNPPTFGQGTKLEIK (SEQ ID NO: 251) (disclosed as SEQ ID NO: 66 in WO 1815). Heavy chain CDR1 1132/1133 has the following amino acid sequence: GFTFSSYA (SEQ ID NO: 252) (disclosed as SEQ ID NO: 13 in WO 2015/091853). Heavy chain CDR2 1132/1133 has the following amino acid sequence: IGSYGGGT (SEQ ID NO: 253) (disclosed as SEQ ID NO: 14 in WO 2015/091853). Heavy chain CDR3 1132/1133 has the following amino acid sequence: ARYVNFGMDY (SEQ ID NO: 254) (disclosed as SEQ ID NO: 15 in WO 2015/091853). Light chain CDR1 1132/1133 has the following amino acid sequence: QSISSY (SEQ ID NO: 255) (disclosed as SEQ ID NO: 16 in WO 2015/091853). Light chain CDR2 1132/1133 has the following amino acid sequence: AAS (SEQ ID NO: 256) (disclosed as SEQ ID NO: 17 in WO 2015/091853). Light chain CDR3 1132/1133 has the following amino acid sequence: QQYGRNPPT (SEQ ID NO: 257) (disclosed as SEQ ID NO: 18 in WO 2015/091853).

The 1107/1108 heavy chain variable region has the following amino acid sequence: EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARRVWGFDYWGQGTLVTVSS (SEQ ID NO: 258) (disclosed as SEQ ID NO: 793/0 2085/0 2085). The light chain variable region 1107/1108 has the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGTDFTLTISSLQPEDFATYYCQQYGVYPFTFGQGTKLEIK (SEQ ID NO: 259) (disclosed as SEQ ID NO: 80 in WO 2015).

Heavy chain CDR1 1107/1108 has the following amino acid sequence: GFTFSSYA (SEQ ID NO: 252) (disclosed as SEQ ID NO: 55 in WO 2015/091853). Heavy chain CDR2 1107/1108 has the following amino acid sequence: ISGSGGST (SEQ ID NO: 260) (disclosed as SEQ ID NO: 56 in WO 2015/091853). Heavy chain CDR3 1107/1108 has the following amino acid sequence: ARRVWGFDY (SEQ ID NO: 261) (disclosed as SEQ ID NO: 57 in WO 2015/091853). Light chain CDR1 1107/1108 has the following amino acid sequence: QSISSY (SEQ ID NO: 255) (disclosed as SEQ ID NO: 58 in WO 2015/091853). Light chain CDR2 1107/1108 has the following amino acid sequence: AAS (SEQ ID NO: 256) (disclosed as SEQ ID NO: 59 in WO 2015/091853). Light chain CDR3 1107/1108 has the following amino acid sequence: QQYGVYPFT (SEQ ID NO: 262) (disclosed as SEQ ID NO: 60 in WO 2015/091853).

In some embodiments, the CD40 agonist is ISF35. ISF35 is a chimeric CD154. ISF is disclosed in PCT Publication WO 2003/099340 and WO 2008/070743.

In some embodiments, the CD40 agonist is dacetuzumab. Dacetuzumab is also known as SGN-40 or huS2C6. Dacetuzumab is a humanized monoclonal antibody specific for CD40. Dacetuzumab is disclosed, for example, in Advani et al. J Clin Oncol. 2009; 27(26):4371-7; and Khubchandani et al. Curr Opin Investig Drugs. 2009; 10(6):579-87.

In some embodiments, the CD40 agonist is lucatumumab (CAS Number: 903512-50-5). Lucatumumab is also known as CHIR-12.12 or HCD-122. Lucatumumab binds to and inhibits CD40, thus inhibiting CD40 ligand-induced cell proliferation and inducing cell lysis via antibody-dependent cellular cytotoxicity (ADCC) in cells overexpressing CD40. Lucatumumab is disclosed, for example, in Tai et al. Cancer Res. 2005; 65(13):5898-906.

Anti-CD40 antibodies can effectively replace helper T cell activity (Ridge, J. et al. (1998) Nature 393: 474-478) and can be used together with anti-PD-1 antibodies (Ito, N. et al. (2000) Immunobiology 201 (5) 527-40).

Examples of OX40 agonists

In one embodiment, the combination described in this application includes an OX40 agonist. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., breast cancer, melanoma, head and neck cancer, or prostate cancer), e.g., hemoblastosis (e.g., lymphomas ( such as B-cell lymphoma.

OX40, also known as CD134, is a cell surface glycoprotein and a member of the tumor necrosis factor receptor (TNF) superfamily, expressed on T lymphocytes and providing a costimulatory signal for proliferation and survival of activated T cells. Activation of OX40 can cause the proliferation of effector T lymphocytes that elicit an immune response against tumor cells that express tumor-associated antigens (TAA).

In some embodiments, the OX40 agonist is selected from mAb 106-222, humanized 106-222 (Hu106), mAb 119-122, or humanized 119-122 (Hu119).

Mab 106-222, humanized 106-222 (Hu106), mAb 119-122, and humanized 119-122 (Hu119) are disclosed, for example, in PCT publication WO 2012/027328 and US Patent 9,006,399. The amino acid sequence of mAb heavy chain variable region 106-222 is disclosed as SEQ ID NO: 4 in WO 2012/027328. The amino acid sequence of the mAb 106-222 light chain variable region is disclosed as SEQ ID NO: 10 in WO 2012/027328. The amino acid sequence of the heavy chain variable region of humanized 106-222 (Hu106) is disclosed as SEQ ID NO: 5 in WO 2012/027328. The light chain variable region amino acid sequence of humanized 106-222 (Hu106) is disclosed as SEQ ID NO: 11 in WO 2012/027328. The amino acid sequence of mAb heavy chain variable region 119-122 is disclosed as SEQ ID NO: 16 in WO 2012/027328. The amino acid sequence of the mAb 119-122 light chain variable region is disclosed as SEQ ID NO: 22 in WO 2012/027328. The amino acid sequence of the heavy chain variable region of humanized 119-122 (Hu119) is disclosed as SEQ ID NO: 17 in WO 2012/027328. The light chain variable region amino acid sequence of humanized 119-122 (Hu119) is disclosed as SEQ ID NO: 23 in WO 2012/027328.

In some embodiments, the OX40 agonist is a humanized monoclonal antibody disclosed in US Pat. No. 7,959,925 and PCT Publication WO 2006/121810.

In some embodiments, the OX40 agonist is selected from MEDI6469, MEDI0562, or MEDI6383. MEDI6469 is a mouse monoclonal antibody against OX40. MEDI0562 is a humanized anti-OX40 monoclonal antibody. MEDI6383 is a monoclonal antibody against OX40.

In some embodiments, an OX40 agonist, such as MEDI6469, is administered intravenously at a dose of about 0.4 mg/kg, such as every other day.

Other examples of antibodies against OX-40 are disclosed, for example, in Weinberg, A. et al. (2000) Immunol 164: 2160-2169.

Examples of CD27 agonists

In one embodiment, the combination described in this application includes a CD27 agonist. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., melanoma, renal cell carcinoma, hormone refractory adenocarcinoma of the prostate, ovarian cancer, breast cancer, adenocarcinoma of the colon and rectum or non-small cell lung cancer), eg, hemoblastosis (eg, lymphomas (eg, Hodgkin's lymphoma, Burkett's lymphoma, mantle cell lymphoma, primary central nervous system lymphoma, or marginal zone B-cell lymphoma) or leukemia (eg, chronic lymphocytic leukemia (CLL) )).

In one embodiment, the CD27 agonist is varlilumab (CAS Number: 1393344-72-3). Varlilumab is also known as CDX-1127 (Celldex) or 1F5. Varlilumab is a fully human monoclonal antibody (mAb) specific for CD27, a molecule in the lymphocyte activation pathway. CDX-1127 is an anti-CD27 agonist mAb that can activate human T cells through T cell receptor stimulation and thus mediate antitumor effects. CDX-1127 may also provide a direct therapeutic effect against tumors expressing CD27.

Varlilumab is disclosed, for example, in Vitale et al., Clin Cancer Res. 2012;18(14):3812-21, WO 2008/051424 and US 8,481,029.

In one embodiment, the CD27 agonist is BION-1402 (BioNovion). BION-1402 is also known as hCD27.15. BION-1402 is a monoclonal antibody against human CD27. BION-1402 can stimulate the proliferation and/or survival of CD27+ cells. BION-1402 can activate human CD27 more efficiently than its CD70 ligand, resulting in a significantly enhanced effect on CD8+ and CD4+ T cell proliferation.

BION-1402 is disclosed for example as hCD27.15 in WO 2012/004367. Это антитело продуцируется гибридомой hCD27.15, которая была депонирована в ATCC 2 июня 2010 года под номером PTA 11008. Вариабельная область тяжелой цепи hCD27.15 имеет аминокислотную последовательность: EVRLQQSGADLVKPGASVKLSCASGFIIKATYMHWVRQRPEQGLEWIGRIDPANGEKYDPKFQVKAITADTSSSTAYLQLNSLTSDDTAVYYCARYAWYFDVWGAGTTVTVSSAKTTPPXVYPXXPGS (SEQ ID NO: 263) (раскрытую как SEQ ID NO: 3 в WO 2012/004367). The hCD27.15 light chain variable region has the amino acid sequence: DIQMTQSPASLSASVGDTVTITCRASENIYSFLAWYHQKQGRSPQLLVYHAKTLAEGVPSRFSGSGSGTQFSLKINSLQAEDFGSYYCQHYYGSPLTFGAGTKLEVKRADAAPTVSIFPPSSEELSL (SEQ ID NO: 264) (disclosed as SEQ ID NO: 302/02/4 in WO02/02/4 in WO02/02/402/402/402/402/302/402/302/302/302/302/4/302/302/302/302/302/302/302/302/302/4/302/302/40). The heavy chain CDR1 of hCD27.15 has the amino acid sequence: GFIIKATYMH (SEQ ID NO: 265) (disclosed as SEQ ID NO: 5 in WO 2012/004367). The heavy chain CDR2 of hCD27.15 has the amino acid sequence: RIDPANGETKYDPKFQV (SEQ ID NO: 266) (disclosed as SEQ ID NO: 6 in WO 2012/004367). The heavy chain CDR3 of hCD27.15 has the amino acid sequence: YAWYFDV (SEQ ID NO: 267) (disclosed as SEQ ID NO: 7 in WO 2012/004367). The light chain CDR1 of hCD27.15 has the amino acid sequence: RASENIYSFLA (SEQ ID NO: 268) (disclosed as SEQ ID NO: 8 in WO 2012/004367). The light chain CDR2 of hCD27.15 has the amino acid sequence: HAKTLAE (SEQ ID NO: 269) (disclosed as SEQ ID NO: 9 in WO 2012/004367). The light chain CDR3 of hCD27.15 has the amino acid sequence: QHYYGSPLT (SEQ ID NO: 270) (disclosed as SEQ ID NO: 10 in WO 2012/004367).

Examples of CSF-1/1R binders

In one embodiment, the combination described in this application includes a CSF-1/1R binding agent. In some embodiments, the combination is used to treat cancer, eg, the cancer described herein, eg, a solid tumor (eg, prostate cancer, breast cancer, or pigmented villonodular synovitis (PVNS)).

In some embodiments, the CSF-1/1R binding agent is a macrophage colony stimulating factor (M-CSF) inhibitor. M-CSF is also sometimes known as CSF-1.

In another embodiment, the CSF-1/1R binding agent is a CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy )-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT Publication WO 2005/073224. In some embodiments, the cancer is selected from brain cancer (eg, glioblastoma multiforme (GBM)), pancreatic cancer, or breast cancer (eg, triple negative breast cancer (TNBC)).

In some embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15) or the compound disclosed in PCT Publication WO 2005/073224 is administered at a dose of 50 mg to 1500 mg, e.g. 75 mg to 1000 mg, 100 mg to 900 mg , 200 mg to 800 mg, 300 mg to 700 mg, 400 mg to 600 mg, 100 mg to 700 mg, 100 mg to 500 mg, 100 mg to 300 mg, 700 mg to 900 mg, 500 mg to 900 mg, 300 mg to 900 mg, 75 mg to 150 mg, 100 mg to 200 mg, 200 mg to 400 mg, 500 mg to 700 mg, or 800 mg to 1000 mg, e.g. at 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg or 1000 mg. In some embodiments, the CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thi-azole -6-yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT Publication WO 2005/073224, is administered daily, for example according to a 7 day/7 day off schedule. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered twice a week, once a week, once every two weeks, once every three weeks, or once at four weeks.

In some embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 50 mg to 150 mg, eg about 100 mg, eg daily, eg according to the administration schedule 7 days / 7 days break. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT Publication WO 2005/073224, is administered at a dose of 100 mg to 200 mg, e.g., about 150 mg, e.g., daily, e.g. 7 days / 7 days break. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 200 mg to 400 mg, for example about 300 mg, for example daily, for example, according to the administration schedule 7 days / 7 days break. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 500 mg to 700 mg, for example about 600 mg, for example daily, for example, according to the administration schedule 7 days / 7 days break. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 800 mg to 1000 mg, for example about 900 mg, for example daily, for example, according to the administration schedule 7 days / 7 days break.

In some embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 50 mg to 150 mg, eg about 100 mg, once a week. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 100 mg to 200 mg, eg about 150 mg, once a week. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 200 mg to 400 mg, eg about 300 mg, once a week. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 500 mg to 700 mg, eg about 600 mg, once a week. In other embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 800 mg to 1000 mg, eg about 900 mg, once a week.

In some embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered orally.

In some embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered in combination with a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule). In one embodiment, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 50 mg to 150 mg (e.g., about 100 mg), e.g., daily (e.g., according to the scheme administration 7 days/7 days break) or once a week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg) , for example, once every 4 weeks, or at a dose of 200 mg to 400 mg (eg, at a dose of 300 mg), for example, once every 3 weeks, for example, by intravenous infusion. In another embodiment, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 100 mg to 200 mg (e.g., about 150 mg), e.g., daily (e.g., according to the scheme administration 7 days/7 days break) or once a week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg) , for example, once every 4 weeks, or at a dose of 200 mg to 400 mg (eg, at a dose of 300 mg), for example, once every 3 weeks, for example, by intravenous infusion. In another embodiment, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 200 mg to 400 mg (e.g., about 300 mg), e.g., daily (e.g., according to the scheme administration 7 days/7 days break) or once a week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg) , for example, once every 4 weeks, or at a dose of 200 mg to 400 mg (eg, at a dose of 300 mg), for example, once every 3 weeks, for example, by intravenous infusion. In another embodiment, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 500 mg to 700 mg (e.g., about 600 mg), e.g., daily (e.g., according to the scheme administration 7 days/7 days break) or once a week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg) , for example, once every 4 weeks, or at a dose of 200 mg to 400 mg (for example, at a dose of 300 mg), for example, once every 3 weeks, for example, by intravenous infusion. In another embodiment, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 800 mg to 1000 mg (e.g., about 900 mg), e.g., daily (e.g., according to the scheme administration 7 days/7 days break) or once a week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg) , for example, once every 4 weeks, or at a dose of 200 mg to 400 mg (for example, at a dose of 300 mg), for example, once every 3 weeks, for example, by intravenous infusion.

In some embodiments, a CSF-1/1R binding agent (e.g., a CSF-1R tyrosine kinase inhibitor), 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat cancer, e.g. tumor (eg, advanced solid tumor), eg, brain cancer (eg, glioblastoma multiforme (GBM), eg, recurrent glioblastoma), breast cancer (eg, triple-negative breast cancer (eg, TNBC)) or pancreatic cancer (eg advanced pancreatic cancer).

In some embodiments, the CSF-1/1R binding agent is an M-CSF inhibitor, Compound A33, or an anti-CSF-1 binding agent disclosed in PCT publication WO 2004/045532 or PCT publication WO 2005/068503, including RX1 or 5H4 ( for example, an antibody molecule or a Fab fragment against M-CSF). In some embodiments, the cancer is selected from endometrial cancer, skin cancer (eg, melanoma), pancreatic cancer, or breast cancer (eg, triple negative breast cancer (TNBC)).

In some embodiments, a CSF-1/1R binding agent, e.g., an M-CSF inhibitor, Compound A33, or a compound disclosed in PCT publication WO 2004/045532 (e.g., an anti-M-CSF antibody molecule or Fab fragment), is administered at an average dose of approximately 10 mg/kg. In some embodiments, the CSF-1/1R binding agent is a CSF1R inhibitor or 4-(2-((1R,2R)-2-hydroxycyclohexylamino)benzothiazol-6-yloxy)-N-methylpicolinamide. 4-(2-((1R,2R)-2-hydroxycyclohexylamino)benzothiazol-6-yloxy)-N-methylpicolinamide is disclosed as Example 157 on page 117 of PCT publication WO 2007/121484.

In some embodiments, the CSF-1/1R binding agent is pexidartinib (CAS Registry Number 1029044-16-3). Pexidartinib is also known as PLX3397 or 5-((5-chloro-1H-pyrrolo[2,3-b]pyridin-3-yl)methyl)-N-((6-(trifluoromethyl)pyridin-3-yl) methyl)pyridine-2-amine. Pexidartinib is a small molecule inhibitor of the receptor tyrosine kinase (RTK) KIT, CSF1R and FLT3. FLT3, CSF1R and FLT3 are overexpressed or mutated in many cancer cell types and play important roles in tumor cell proliferation and metastasis. PLX3397 can bind to and inhibit phosphorylation of stem cell factor receptor (KIT), colony stimulating factor receptor 1 (CSF1R), and FMS-like tyrosine kinase 3 (FLT3), which can lead to inhibition of tumor cell proliferation and downmodulation of macrophages, osteoclasts, and mast cells involved in osteolytic metastatic disease. In some embodiments, a CSF-1/1R binding agent, eg, pexidartinib, is used in combination with a PD-1 inhibitor, eg, an anti-PD-1 antibody molecule described herein.

In some embodiments, the CSF-1/1R binder is emactuzumab. Emactuzumab is also known as RG7155 or RO5509554. Emactuzumab is a humanized IgG1 mAb specific for CSF1R. In some embodiments, a CSF-1/1R binding agent, eg, pexidartinib, is used in combination with a PD-L1 inhibitor, eg, an anti-PD-L1 antibody molecule described herein.

In some embodiments, the CSF-1/1R binding agent is FPA008. FPA008 is a humanized mAb that inhibits CSF1R. In some embodiments, a CSF-1/1R binding agent, such as FPA008, is used in combination with a PD-1 inhibitor, such as an anti-PD-1 antibody molecule described herein.

Examples of IL-17 Inhibitors

In one embodiment, the combination described in this application includes an inhibitor of interleukin-17 (IL-17). In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor, e.g., breast cancer (e.g., triple negative breast cancer (TNBC), lung cancer (e.g., non-small cell lung cancer ( NSCLC)) or colon cancer In some embodiments, the cancer is selected from colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS CRC)), lung cancer (e.g., non-small cell lung cancer), or breast cancer ( e.g. triple negative lung cancer (TNBC)).

In some embodiments, the IL-17 inhibitor is secukinumab (CAS Registry Number: 875356-43-7 (heavy chain) and 875356-44-8 (light chain)). ‎Secukinumab is also known as AIN457 and Cosentyx®. Secukinumab is a recombinant human monoclonal IgG1/κ antibody that specifically binds to IL-17A. It is expressed in a recombinant Chinese hamster ovary (CHO) cell line.

Секукинумаб описан, например, в WO 2006/013107, US 7,807,155, US 8,119,131, US 8,617,552 и EP 1776142. Вариабельная область тяжелой цепи секукинумаба имеет следующую аминокислотную последовательность: EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYWMNWVRQAPGKGLEWVAAINQDGSEKYYVGSVKGRFTISRDNAKNSLYLQMNSLRVEDTAVYYCVRDYYDILTDYYIHYWYFDLWGRGTLVTVSS (SEQ ID NO: 271) (раскрытую как SEQ ID NO: 8 в WO 2006/013107). The light chain variable region of secukinumab has the following amino acid sequence: EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSPCTFGQGTRLEIKR (SEQ ID NO: 298) (disclosed as SEQ ID NO: 10 in WO 2006/013107). The heavy chain CDR1 of secukinumab has the amino acid sequence NYWMN (SEQ ID NO: 272) (disclosed as SEQ ID NO: 1 in WO 2006/013107). The secukinumab heavy chain CDR2 has the amino acid sequence AINQDGSEKYYVGSVKG (SEQ ID NO: 273) (disclosed as SEQ ID NO: 2 in WO 2006/013107). The heavy chain CDR3 of secukinumab has the amino acid sequence DYYDILTDYYIHYWYFDL (SEQ ID NO: 274) (disclosed as SEQ ID NO: 3 in WO 2006/013107). The light chain CDR1 of secukinumab has the amino acid sequence RASQSVSSSYLA (SEQ ID NO: 275) (disclosed as SEQ ID NO: 4 in WO 2006/013107). The secukinumab light chain CDR2 has the amino acid sequence GASSRAT (SEQ ID NO: 276) (disclosed as SEQ ID NO: 5 in WO 2006/013107). The secukinumab light chain CDR3 has the amino acid sequence GASSRAT (SEQ ID NO: 299) (disclosed as SEQ ID NO: 6 in WO 2006/013107).

In some embodiments, the IL-17 inhibitor is CJM112. CJM112 is also known as XAB4. CJM112 is a fully human monoclonal antibody (eg, IgG1/κ isotype) specific for IL-17A.

CJM112 is disclosed, for example, in WO 2014/122613. Тяжелая цепь CJM112 имеет следующую аминокислотную последовательность: EVQLVESGGDLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARDRGSLYYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 277) (раскрытую как SEQ ID NO: 14 в WO 2014/122613). Легкая цепь CJM112 имеет следующую аминокислотную последовательность: AIQLTQSPSSLSASVGDRVTITCRPSQGINWELAWYQQKPGKAPKLLIYDASSLEQGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 278) (раскрытую как SEQ ID NO: 44 в WO 2014/122613).

CJM112 can bind to human, cynomolgus, mouse, and rat IL-17A and neutralize the bioactivity of these cytokines in vitro and in vivo . IL-17A, a member of the IL-17 family, is a major pro-inflammatory cytokine that has been shown to play important roles in many immune-mediated conditions such as psoriasis and cancer (Witowski et al. (2004) Cell Mol. Life Sci. p 567-79; Miossec and Kolls (2012) Nat. Rev. Drug Discov. pp. 763-76). Without being limited by theory, it is believed that in some embodiments, an IL-17 inhibitor, e.g., an anti-IL17 antibody, can enhance the PD-1-mediated response, e.g., by blocking IL-17 mediated proliferation of suppressor T-cell neutrophils and/or reducing metastasis ( Coffelt et al (2015) Nature p.345-8).

In some embodiments, the combination described herein includes an IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD- 1).

IL-17 is an important mediator of innate immunity and chronic inflammatory response. For example, by causing increased production of chemokines in various tissues, IL-17 is important in the pathophysiology of an autoimmune disease (Isailovic et al. (2015) J. Autoimmun. p. 1-11). IL-17 is secreted primarily by CD4+ Th17, but also by γδ T cells, CD8+ T cells, neutrophils and eosinophils, and it recruits monocytes and neutrophils to the site of inflammation (Alshaker and Matalka (2011) Cancer Cell Int. p. 11 -33). The role of Th17 cell infiltrate in tumors has been investigated in various types of cancers and has been correlated with poor prognosis in many settings (Zeng et al. (2015) Int. J. Clin. Exp. Med. p. 10515-36). Th17 cells and, in particular, IL-17, perform both antitumor and protumor functions. Under certain conditions, IL-17 induces tumor vascularization, cell proliferation, tumorigenesis, metastasis, and/or recruitment of macrophage-derived suppressor cells into the tumor microenvironment (Yang et al. (2014) Mediators Inflamm. p. 014:623759). There is a method of targeting IL-17 in the treatment of malignancy (Zou and Restifo (2010) Nat. Rev. Immunol. p. 246-56). In some embodiments, an IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107 is increased or used to increase a PD-1 mediated response, e.g. block IL-17 mediated proliferation of suppressor T-cell neutrophils and/or reduce metastasis .

In some embodiments, an IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), are dosed at and/ or according to a temporal scheme, which in combination provide the desired antitumor activity.

In one embodiment, the IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107 is administered at a dose of 5 mg to 100 mg, e.g., 10 mg to 75 mg, 15 mg to 50 mg, 20 mg up to 30 mg, from 10 mg to 25 mg, or from 25 mg to 40 mg, for example, at a dose of 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg, such as once every two weeks, every four weeks, once every six weeks, or once every eight weeks, such as intravenously.

In one embodiment, the IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107 is administered at a dose of 20 mg to 30 mg (eg 25 mg), eg once every four weeks. In some embodiments, an IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107 is administered intravenously. In one embodiment, the IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107, is administered at a dose of 20 mg to 30 mg (e.g., 25 mg), e.g., once every four weeks, e.g., by intravenous infusion , and a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (eg, a 400 mg dose), eg, once every 4 weeks, eg, by intravenous infusion.

In one embodiment, the IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107 is administered at a dose of 5 mg to 750 mg, e.g., 10 mg to 500 mg, 15 mg to 450 mg, 20 mg up to 400 mg, 25 to 350 mg, 50 mg to 300 mg, 75 mg to 250 mg, 100 mg to 200 mg, 25 mg to 75 mg, or 40 mg to 60 mg, e.g. 5 mg, 10 mg, 15 mg, 20 mg, 30 mg, 40 mg, 50 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg , 450 mg or 500 mg, for example, once every two weeks, once every four weeks, once every six weeks, or once every eight weeks, for example, subcutaneously.

In one embodiment, the IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107 is administered at a dose of 40 mg to 60 mg (eg 50 mg), eg once every four weeks. In some embodiments, an IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107 is administered subcutaneously. In one embodiment, the IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107, is administered at a dose of 40 mg to 60 mg (e.g., 50 mg), e.g., once every four weeks, e.g., by subcutaneous injection , and a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (eg, a 400 mg dose), eg, once every 4 weeks, eg, by intravenous infusion.

In one embodiment, an IL-17 inhibitor, CJM112, or a compound disclosed in WO 2006/013107, is used in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat colon and rectal cancer (CRC), lung cancer (eg, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple-negative breast cancer (TNBC)).

In some embodiments, the IL-17 inhibitor is ixekizumab (CAS Number: 1143503-69-8). ‎Ixekizumab is also known as LY2439821. Ixekizumab is a humanized IgG4 monoclonal antibody specific for IL-17A.

Ixekizumab is described, for example, in WO 2007/070750, US 7,838,638 and US 8,110,191. The heavy chain variable region of ixekizumab has the following amino acid sequence: QVQLVQSGAEVKKPGSSVKVSCKASGYSFTDYHIHWVRQAPGQGLEWMGVINPMYGTTDYNQRFKGRVTITADESTSTAYMELSSLRSEDTAVYYCARYDYFTGTGVYWGQGTLVTVSS (SEQ ID NO: 279) (disclosed as SEQ ID NO: 2070/070/2070/2070/2070/2070/2070/2070). The light chain variable region of ixekizumab has the following amino acid sequence: DIVMTQTPLSLSVTPGQPASISCRSSRSLVHSRGNTYLHWYLQKPGQSPQLLIYKVSNRFIGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHLPFTFGQGTKLEIK (SEQ ID NO: 280) (disclosed as SEQ ID NO: 241 in WO 20707/0).

In some embodiments, the IL-17 inhibitor is brodalumab (CAS Number: 1174395-19-7). Brodalumab is also known as AMG 827 or AM-14. Brodalumab binds to the interleukin-17 receptor (IL-17RA) and blocks the activation of the IL-17 receptor.

Brodalumab is disclosed, for example, in WO 2008/054603, US 7,767,206, US 7,786,284, US 7,833,527, US 7,939,070, US 8,435,518, US 8,545,842, US 8,790,648 and US 9,073,999. The heavy chain CDR1 of brodalumab has the amino acid sequence RYGIS (SEQ ID NO: 281) (disclosed as SEQ ID NO: 146 in WO 2008/054603). The heavy chain CDR2 of brodalumab has the amino acid sequence WISTYSGNTNYAQKLQG (SEQ ID NO: 282) (disclosed as SEQ ID NO: 147 in WO 2008/054603). The heavy chain CDR3 of brodalumab has the amino acid sequence RQLYFDY (SEQ ID NO: 283) (disclosed as SEQ ID NO: 148 in WO 2008/054603). The light chain CDR1 of brodalumab has the amino acid sequence RASQSVSSNLA (SEQ ID NO: 284) (disclosed as SEQ ID NO: 224 in WO 2008/054603). The heavy chain CDR2 of brodalumab has the amino acid sequence DASTRAT (SEQ ID NO: 285) (disclosed as SEQ ID NO: 225 in WO 2008/054603). The heavy chain CDR3 of brodalumab has the amino acid sequence QQYDNWPLT (SEQ ID NO: 286) (disclosed as SEQ ID NO: 226 in WO 2008/054603).

Examples of IL-1β inhibitors

In one embodiment, the combination described in this application includes an inhibitor of interleukin-1 beta (IL-1β). In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., hemoblastosis (e.g., lymphomas (e.g., Hodgkin's lymphomas), leukemia (e.g., acute or chronic leukemia), or multiple myeloma) or solid tumor (e.g., colon and rectal cancer (CLC), lung cancer (e.g., non-small cell lung cancer (NSCLC)) or breast cancer (e.g., triple negative breast cancer (TNBC)). In some embodiments, the cancer is selected from colon cancer. and rectum (eg, microsatellite stable colon and rectal cancer (MSS CRC), lung cancer (eg, non-small cell lung cancer), or breast cancer (eg, triple negative lung cancer (TNBC)).

In some embodiments, the IL-1β inhibitor is canakinumab. Canakinumab is also known as ACZ885 or Ilaris®. Canakinumab is a human monoclonal IgG1/κ antibody that neutralizes the bioactivity of human IL-1β.

Канакинумаб раскрыт, например, в WO 2002/16436, US 7,446,175, и EP 1313769. Вариабельная область тяжелой цепи канакинумаба имеет следующую аминокислотную последовательность: MEFGLSWVFLVALLRGVQCQVQLVESGGGVVQPGRSLRLSCAASGFTFSVYGMNWVRQAPGKGLEWVAIIWYDGDNQYYADSVKGRFTISRDNSKNTLYLQMNGLRAEDTAVYYCARDLRTGPFDYWGQGTLVTVSS (SEQ ID NO: 287) (раскрытую как SEQ ID NO: 1 в US 7,446,175). The canakinumab light chain variable region has the following amino acid sequence: MLPSQLIGFLLLWVPASRGEIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLIKYASQSFSGVPSRFSGSGTDFTLTINSLEAEDAAAYYCHQSSSLPFTFGPGTKVDIK (SEQ ID NO: 288) (disclosed as SEQ ID NO: 2 in US 7,446).

Canakinumab has been used, for example, for the treatment of cryopyrin-associated periodic syndromes (CAPS) in adults and children, for the treatment of systemic juvenile idiopathic arthritis (SJIA), for the symptomatic treatment of attacks of acute gouty arthritis in adults, and for the treatment of other IL-1β mediated inflammatory diseases. Without being limited by theory, it is believed that in some embodiments, IL-1β inhibitors, such as canakinumab, can enhance the antitumor immune response, for example, by blocking one or more functions of IL-1β, including, for example, the recruitment of immunosuppressive neutrophils to the tumor microenvironment, stimulation of tumor angiogenesis and/or stimulation of metastasis (Dinarello (2010) Eur. J. Immunol. p. 599-606). In some embodiments, the combination described herein comprises an IL-1β inhibitor, canakinumab, or a compound disclosed in WO 2002/16436, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD- 1).

IL-1 is a secreted pleiotropic cytokine with a major role in inflammation and immune response. An increase in IL-1 has been observed in various clinical settings, including cancer (Apte et al. (2006) Cancer Metastasis Rev. p. 387-408; Dinarello (2010) Eur. J. Immunol. p. 599-606). IL-1β is elevated in lung, breast, and colon and rectal cancers (Voronov et al. (2014) Front Physiol. p. 114), and is associated with poor prognosis (Apte et al. (2000) Adv. Exp. Med. Biol. pp. 277-88). Without being limited by theory, it is believed that in some embodiments, IL-1β, derived from the tumor microenvironment and secreted by malignant cells, induces tumor cell proliferation, increases invasiveness, and attenuates the antitumor immune response, in part as a result of the recruitment of inhibitory neutrophils (Apte et al. (2006) ) Cancer Metastasis Rev. pp. 387-408; Miller et al. (2007) J. Immunol. pp. 6933-42). Experimental data indicate that inhibition of IL-1β leads to a reduction in tumor burden and metastasis (Voronov et al. (2003) Proc. Natl. Acad. Sci. U.S.A. p. 2645-50). Canakinumab can bind IL-1β and inhibit IL-1 mediated signaling. Thus, in some embodiments, an IL-1β inhibitor, such as canakinumab, enhances or is used to enhance the immune-mediated antitumor effect of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule).

In some embodiments, an IL-1β inhibitor, canakinumab, or a compound disclosed in WO 2002/16436, and an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), are dosed at and/ or according to a temporal scheme, which in combination provide the desired antitumor activity.

In one embodiment, the IL-1β inhibitor, canakinumab, or a compound disclosed in WO 2002/16436, is administered at a dose of 25 mg to 200 mg, e.g., 50 mg to 150 mg, 80 mg to 120 mg, between 140 mg and 160 mg, 40 mg to 60 mg, 80 mg to 100 mg, 50 mg to 100 mg, or 100 mg to 150 mg, such as 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, or 200 mg, for example, once every four weeks, one once every six weeks, once every eight weeks, once every ten weeks, or once every twelve weeks.

In one embodiment, the IL-1β inhibitor, canakinumab, or a compound disclosed in WO 2002/16436, is administered at a dose of 80 mg to 120 mg (eg, 100 mg dose), for example, once every eight weeks. In some embodiments, the IL-1β inhibitor, canakinumab, or a compound disclosed in WO 2002/16436 is administered subcutaneously, for example, in the abdomen or thigh. In one embodiment, an IL-1β inhibitor, e.g., canakinumab, is administered at a dose of 80 mg to 120 mg (e.g., 100 mg dose), e.g., once every eight weeks, e.g., by subcutaneous injection, and a PD-1 inhibitor (eg, anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (eg, 400 mg), eg, once every 4 weeks, eg, by intravenous infusion.

In some embodiments, an IL-1β inhibitor, canakinumab, or a compound disclosed in WO 2002/16436, is used in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat colon and rectal cancer (e.g., MSS CRC), lung cancer (eg non-small cell lung cancer (NSCLC) or breast cancer (eg triple negative breast cancer (TNBC)).

Examples of CXCR2 inhibitors

In one embodiment, the combination described herein comprises a chemokine 2 (CXCR2) receptor inhibitor (C-X-C motif) inhibitor. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor, e.g., breast cancer, metastatic sarcoma, pancreatic cancer, melanoma, renal cell carcinoma (RCC), non-small cell lung cancer (NSCLC) or a pediatric tumor (eg, rhabdomyosarcoma).

In some embodiments, the CXCR2 inhibitor is danirixin (CAS Number: 954126-98-8). Danirixin is also known as GSK1325756 or 1-(4-chloro-2-hydroxy-3-piperidin-3-ylsulfonylphenyl)-3-(3-fluoro-2-methylphenyl)urea. Danirixin is disclosed, for example, in Miller et al. Eur J Drug Metab Pharmacokinet (2014) 39:173-181; and Miller et al. BMC Pharmacology and Toxicology (2015), 16:18.

In some embodiments, the CXCR2 inhibitor is reparixin (CAS Number: 266359-83-5). Reparixin is also known as repertaxin or (2R)-2-[4-(2-methylpropyl)phenyl]-N-methylsulfonylpropanamide. Reparixin is a noncompetitive allosteric inhibitor of CXCR1/2. Reparixin is disclosed, for example, in Zarbock et al. British Journal of Pharmacology (2008), 1-8.

In some embodiments, the CXCR2 inhibitor is navarixin. Navarixin is also known as MK-7123, SCH 527123, PS291822 or 2-hydroxy-N,N-dimethyl-3-[[2-[[(1R)-1-(5-methylfuran-2-yl)propyl]amino] -3,4-dioxocyclobuten-1-yl]amino]benzamide. Navarixin is disclosed, for example, in Ning et al. Mol Cancer Ther. 2012;11(6):1353-64.

Examples of PI3K-γ, -δ inhibitors

In one embodiment, the combination described herein comprises a phosphatidylinositol 4,5-bisphosphate 3-kinase (PI3K) inhibitor, such as phosphatidylinositol 4,5-bisphosphate-3-kinase gamma and/or delta (PI3K-γ ,δ). In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., prostate cancer, breast cancer, brain cancer, bladder cancer, pancreatic cancer, kidney cancer, solid tumor, liver cancer, non-small cell lung cancer, endocrine cancer, ovarian cancer, melanoma, cancer of the female reproductive system, digestive system/gastrointestinal tract cancer, glioblastoma multiforme, head and neck cancer, or colon cancer), such as hemoblastosis (eg, leukemia (eg, lymphocytic leukemia, eg, chronic lymphocytic leukemia (CLL) (eg, recurrent CLL)) eg, lymphomas (eg, non-Hodgkin's lymphoma (eg, recurrent follicular B-cell non-Hodgkin's lymphoma (FL) or recurrent small lymphocytic lymphoma ( SLL)), or for example, multiple myeloma).

In some embodiments, the PI3K inhibitor is an inhibitor of the delta and gamma isoforms of PI3K. Examples of PI3K inhibitors that can be used in combination are described, for example, in WO 2010/036380, WO 2010/006086, WO 09/114870, WO 05/113556, GSK 2126458, GDC-0980, GDC-0941, Sanofi XL147, XL756 , XL147, PF-46915032, BKM 120, CAL-101, CAL 263, SF1126, PX-886, and a dual PI3K inhibitor (eg, Novartis BEZ235).

In some embodiments, the PI3K-γ,δ inhibitor is idelicib (CAS Number: 870281-82-6). Idelalisib is also known as ZYDELIG®, GS-1101, CAL-101, or 5-fluoro-3-phenyl-2-[(1S)-1-(7H-purin-6-ylamino)propyl]-4(3H)-quinazolinone . Idelalisib blocks P110δ, the delta isoform of PI3K. Idelalisib is disclosed, for example, in Wu et al. Journal of Hematology & Oncology (2013) 6:36.

In some embodiments, the PI3K-γ,δ inhibitor is dactolisib (Compound A4) or 8-(6-methoxy-pyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3-trifluoromethyl- phenyl)-1,3-dihydro-imidazo[4,5-c]quinolin-2-one (Compound A41), or the compound disclosed in PCT Publication WO 2006/122806.

In some embodiments, the PI3K-γ,δ inhibitor is buparlisib (Compound A6) or a compound disclosed in PCT publication WO 2007/084786.

In one embodiment, a PI3K-γ,δ inhibitor such as buparlisib (Compound A6) or a compound disclosed in PCT publication WO 2007/084786 is administered at a dose of approximately 100 mg (eg per day).

Other examples of PI3K-γ,δ inhibitors that can be used in combination include, for example, pictilisib (GDC-0941), LY294002, pilaralisib (XL147), PI-3065, PI-103, VS-5584 (SB2343), CZC24832, duvelisib (IPI-145, INK1197), TG100-115, CAY10505, GSK1059615, PF-04691502, AS-605240, voxtalisib (SAR245409, XL765), IC-87114, omipalisib (GSK2126458, GSK4458), TG1007 , PKI-587), PKI-402, XL147 analog, PIK-90, PIK-293, PIK-294, 3-methyladenine (3-MA), AS-252424, AS-604850, or apitolisib (GDC-0980, RG7422) .

In some embodiments, the PI3K inhibitor is Compound A8 or the compound described in PCT Publication WO2010/029082.

In some embodiments, the PI3K inhibitor is a pan PI3K inhibitor, (4S, 5R)-3-(2'-amino-2-morpholino-4'-(trifluoromethyl)-[4,5'-bipyrimidin]-6-yl) -4-(hydroxymethyl)-5-methyloxazolidin-2-one (Compound A13) or the compound disclosed in PCT Publication WO2013/124826.

Examples of PI3K-γ,-δ inhibitors include, but are not limited to, duvelisib and idelicib. Idelalisib (also called GS-1101 or CAL-101; Gilead) is a small molecule that blocks the PI3K delta isoform. The structure of Idelalisib (5-fluoro-3-phenyl-2-[(1S)-1-(7H-purin-6-ylamino)propyl]-4(3H)-quinazolinone) is shown below.

Duvelysib (also called IPI-145; Infinity Pharmaceuticals and Abbvie) is a small molecule that blocks PI3K-δ,γ. The structure of duvelysib (8-chloro-2-phenyl-3-[(1S)-1-(9H-purin-6-ylamino)ethyl]-1(2H)-isoquinolinone) is shown below.

In one embodiment, the inhibitor is a dual phosphatidylinositol 3-kinase (PI3K) and mTOR inhibitor selected from 2-amino-8-[trans-4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxy-3-pyridinyl )-4-methyl-pyrido[2,3-d]pyrimidin-7(8H)-one (PF-04691502); N-[4-[[4-(dimethylamino)-1-piperidinyl]carbonyl]phenyl]-N'-[4-(4,6-di-4-morpholinyl-1,3,5-triazine-2 -yl)phenyl]urea (PF-05212384, PKI-587); 2-methyl-2-{4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl] phenyl}propanenitrile (BEZ-235); apitolisib (GDC-0980, RG7422); 2,4-difluoro-N-{2-(methyloxy)-5-[4-(4-pyridazinyl)-6-quinolinyl]-3-pyridinyl}benzenesulfonamide (GSK2126458); 8-(6-methoxypyridin-3-yl)-3-methyl-1-(4-(piperazin-1-yl)-3-(trifluoromethyl)phenyl)-1H-imidazo[4,5-c]quinoline- 2(3H)-one maleic acid (NVP-BGT226); 3-[4-(4-morpholinyl-pyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol (PI-103); 5-(9-isopropyl- 8-methyl-2-morpholino-9H-purin-6-yl)pyrimidin-2-amine (VS-5584, SB2343); or N-[2-[(3,5-dimethoxyphenyl)amino]quinoxalin-3-yl ]-4-[(4-methyl-3-methoxyphenyl)carbonyl]aminophenylsulfonamide (XL765).

Examples of BAFF-R Inhibitors

In one embodiment, the combination described herein comprises a B cell activating factor receptor (BAFF-R) inhibitor. In some embodiments, the combination is used to treat cancer, eg, the cancer described herein, eg, hemoblastosis, eg, leukemia (eg, chronic lymphocytic leukemia (CLL), eg, relapsed or refractory chronic lymphocytic leukemia).

In one embodiment, the BAFF-R inhibitor is VAY736. VAY736 is a fully human anti-BAFF-R monoclonal antibody derived from the combinatorial antibody library (HuCAL). BAFF-R, also known as a member of the tumor necrosis factor 13C receptor superfamily, is overexpressed in some types of tumor cells and in some autoimmune diseases. VAY736 has anti-inflammatory and antitumor activity. In cancer cells, BAFF-R plays an important role in B cell proliferation and survival. VAY736 targets and binds to BAFF-R, resulting in inhibition of BAFF/BAFF-R interaction and BAFF-R-mediated signaling. This may reduce the growth of tumor cells expressing BAFF-R.

VAY736 is disclosed, for example, in US 8,106,163. The heavy chain CDR1 of VAY736 has the amino acid sequence GDSVSSNSAAWG (SEQ ID NO: 289) (disclosed as SEQ ID NO: 3 in US 8,106,163). The heavy chain CDR2 of VAY736 has the amino acid sequence RIYYRSKWYNSYAVSVKS (SEQ ID NO: 290) (disclosed as SEQ ID NO: 10 in US 8,106,163). The heavy chain CDR3 of VAY736 has the amino acid sequence YDWVPKIGVFDS (SEQ ID NO: 300) (disclosed as SEQ ID NO: 17 in US 8,106,163). The VAY736 light chain CDR1 has the amino acid sequence RASQFISSSYLS (SEQ ID NO: 291) (disclosed as SEQ ID NO: 24 in US 8,106,163). The VAY736 light chain CDR2 has the amino acid sequence LLIYGSSSRAT (SEQ ID NO: 292) (disclosed as SEQ ID NO: 31 in US 8,106,163). The VAY736 light chain CDR3 has the amino acid sequence QQLYSSPM (SEQ ID NO: 293) (disclosed as SEQ ID NO: 38 in US 8,106,163). The VAY736 heavy chain variable region has the following amino acid sequence: QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWGWIRQSPGRGLEWLGRIYYRSKWYNSYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARYDWVPKIGVFDSWGQGTLVTVSS (SEQ ID NO: 294) (disclosed as SEQ ID NO: 36, 36 in US 8.36). The VAY736 light chain variable region has the following amino acid sequence: DIVLTQSPATLSLSPGERATLSCRASQFISSSYLSWYQQKPGQAPRLLIYGSSSRATGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQLYSSPMTFGQGTKVEIKRT (SEQ ID NO: 295) (disclosed as SEQ ID NO: 45 in US 8,106,163). Тяжелая цепь VAY736 имеет следующую аминокислотную последовательность: QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWGWIRQSPGRGLEWLGRIYYRSKWYNSYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCARYDWVPKIGVFDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 296) (раскрытую как SEQ ID NO: 75 в US 8,106,163). Вариабельная область легкой цепи VAY736 имеет следующую аминокислотную последовательность: DIVLTQSPATLSLSPGERATLSCRASQFISSSYLSWYQQKPGQAPRLLIYGSSSRATGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQLYSSPMTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 297) (раскрытую как SEQ ID NO: 71 в US 8,106,163).

Examples of MALT-1/BTK Inhibitors

In one embodiment, the combination described in this application includes an inhibitor of MALT-1 and/or BTK. In some embodiments, the combination is used to treat cancer, such as the cancer described herein.

Examples of MALT-1/BTK inhibitors include, but are not limited to, (S)-1-(6-(2H-1,2,3-triazol-2-yl)-5-(trifluoromethyl)pyridin-3-yl)- 3-(2-chloro-7-(1-methoxyethyl)pyrazolo[1,5-a]pyrimidin-6-yl)urea, (S)-1-(2-chloro-7-(1-methoxyethyl)pyrazolo[ 1,5-a]pyrimidin-6-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea, (S)-1-(2-chloro-7-(1-methoxyethyl)pyrazolo[1 ,5-a]pyrimidin-6-yl)-3-(1-methyl-2-oxo-5-(trifluoromethyl)-1,2-dihydropyridin-3-yl)urea, (R)-1-(6- (2H-1,2,3-triazol-2-yl)-5-(trifluoromethyl)pyridin-3-yl)-3-(2-chloro-7-(1-methoxy-2-methylpropyl)pyrazolo[1,5 -a]pyrimidin-6-yl)urea, (R)-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2 -chloro-7-(1-methoxy-2-methylpropyl)pyrazolo[1,5-a]pyrimidin-6-yl)urea, (S)-1-(7-(1-methoxyethyl)-2-methylpyrazolo[1 ,5-a]pyrimidin-6-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea, (S)-1-(2-fluoro-7-(1-methoxyethyl)pyrazolo[1, 5-a]pyrimidin-6-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea and (S)-1-(2-chloro-7-(1-methoxyethyl)pyrazolo[1,5 -a]pyrimidin-6-yl)-3-(5-cyanopyridin-3- silt) urea.

Examples of BTK inhibitors include, but are not limited to, ibrutinib (PCI-32765); GDC-0834; RN-486; CGI-560; CGI-1764; HM-71224; CC-292; ONO-4059; CNX-774 or LFM-A13. In one embodiment, the BTK inhibitor does not reduce or inhibits the kinase activity of interleukin-2-inducible kinase (ITK), for example, selected from GDC-0834; RN-486; CGI-560; CGI-1764; HM-71224; CC-292; ONO-4059; CNX-774 or LFM-A13.

In one embodiment, the kinase inhibitor is a BTK inhibitor, such as ibrutinib (PCI-32765). Structure of ibrutinib (1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]prop-2 -en-1-she) is shown below.

In other embodiments, the BTK inhibitor is the BTK inhibitor described in WO/2015/079417, which is hereby incorporated by reference in its entirety. For example, in some embodiments, the BTK inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof;

wherein

R1 is hydrogen, C1-C6 alkyl which is optionally replaced by hydroxy;

R2 is hydrogen or halogen;

R3 is hydrogen or halogen;

R4 is hydrogen;

R5 is hydrogen or halo;

or R4 and R5 are attached to each other and represent a bond, -CH2-, -CH2-CH2-, -CH=CH-, -CH=CH-CH2-; -CH2-CH=CH-; or -CH2-CH2-CH2-;

R6 and R7 are independently H, C1-C6, alkyl which is optionally substituted with hydroxyl, C3-C6 cycloalkyl which is optionally substituted with halogen or hydroxy, or halogen;

R8, R9, R, R', R10 and R11 are independently H or C1-C6 alkyl which is optionally substituted with C1-C6 alkoxy; or any two of R8, R9, R, R', R10 and R11, together with the carbon atom to which they are attached, may form a 3-6 membered saturated carbocyclic ring;

R12 is hydrogen or C1-C6 alkyl which is optionally substituted with halo or C1-C6 alkoxy;

or R12 and any of R8, R9, R, R', R10 or R11 together with the atoms to which they are attached may form a 4, 5, 6 or 7 membered azacyclic ring which may optionally be substituted with halogen, cyano, hydroxyl , C1-C6 alkyl or C1-C6 alkoxy;

n is 0 or 1; and

R13 is C2-C6 alkenyl which is optionally substituted with C1-C6 alkyl, C1-C6 alkoxy or N,N-di-C1-C6 alkylamino; C2-C6 alkynyl which is optionally substituted with C1-C6 alkyl or C1-C6 alkoxy; or C2-C6 alkylene oxide, which is optionally substituted with C1-C6 alkyl.

In some embodiments, the BTK inhibitor of Formula I is selected from the following: N-(3-(5-((1-acryloylazetidin-3-yl)oxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl) -4-cyclopropyl-2-fluorobenzamide; (E)-N-(3-(6-amino-5-((1-(but-2-enoyl)azetidin-3-yl)oxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl) -4-cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-((1-propioylazetidin-3-yl)oxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-((1-(but-2-inoyl)azetidin-3-yl)oxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl -2-fluorobenzamide; N-(3-(5-((1-acryloylpiperidin-4-yl)oxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-(2-(N-methylacrylamido)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; (E)-N-(3-(6-amino-5-(2-(N-methylbut-2-enamido)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl- 2-fluorobenzamide; N-(3-(6-amino-5-(2-(N-methylpropiolamido)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; (E)-N-(3-(6-amino-5-(2-(4-methoxy-N-methylbut-2-enamido)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)- 4-cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-(2-(N-methylbut-2-ynamido)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; N-(2-((4-amino-6-(3-(4-cyclopropyl-2-fluorobenzamido)-5-fluoro-2-methylphenyl)pyrimidin-5-yl)oxy)ethyl)-N-methyloxiran-2 -carboxamide; N-(2-((4-amino-6-(3-(6-cyclopropyl-8-fluoro-1-oxoisoquinolin-2-(1H)-yl)phenyl)pyrimidin-5-yl)oxy)ethyl )-N-methylacrylamide; N-(3-(5-(2-acrylamido-ethoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-(2-(N-ethylacryl-amido)ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-(2-(N-(2-fluoroethyl)acrylamido)-ethoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2- fluorobenzamide; N-(3-(5-(1-acrylamidocyclopropyl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; (S)-N-(3-(5-(2-acrylamidopropoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; (S)-N-(3-(6-amino-5-(2-(but-2-ynamido)propoxy)-pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2 -fluorobenzamide; (S)-N-(3-(6-amino-5-(2-(N-methylacrylamido)-propoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide ; (S)-N-(3-(6-amino-5-(2-(N-methylbut-2-ynamido)-propoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl -2-fluorobenzamide; N-(3-(6-amino-5-(3-(N-methylacrylamido)propoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; (S)-N-(3-(5-((1-acryloylpyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide ; (S)-N-(3-(6-amino-5-((1-(but-2-inoyl)pyrrolidin-2-yl)methoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl) -4-cyclopropyl-2-fluorobenzamide; (S)-2-(3-(5-((1-acryloylpyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-(hydroxymethyl)phenyl)-6-cyclopropyl- 3,4-dihydroisoquinoline-1(2H)-one; N-(2-((4-amino-6-(3-(6-cyclopropyl-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)-5-fluoro-2-(hydroxymethyl)phenyl) pyrimidin-5-yl)oxy)ethyl)-N-methylacrylamide; N-(3-(5-(((2S,4R)-1-acryloyl-4-methoxypyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4 -cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-(((2S,4R)-1-(but-2-inoyl)-4-methoxypyrrolidin-2-yl)methoxy)pyrimidin-4-yl)-5-fluoro -2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; 2-(3-(5-(((2S,4R)-1-acryloyl-4-methoxypyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-(hydroxymethyl)phenyl )-6-cyclopropyl-3,4-dihydroisoquinolin-1(2H)-one; N-(3-(5-(((2S,4S)-1-acryloyl-4-methoxypyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4 -cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-(((2S,4S)-1-(but-2-inoyl)-4-methoxypyrrolidin-2-yl)methoxy)pyrimidin-4-yl)-5-fluoro -2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; N-(3-(5-(((2S,4R)-1-acryloyl-4-fluoropyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4 -cyclopropyl-2-fluorobenzamide; N-(3-(6-amino-5-(((2S,4R)-1-(but-2-inoyl)-4-fluoropyrrolidin-2-yl)methoxy)pyrimidin-4-yl)-5-fluoro -2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide; (S)-N-(3-(5-((1-Acryloylazetidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide ; (S)-N-(3-(6-amino-5-((1-propioylazetidin-2-yl)methoxy)pyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2- fluorobenzamide; (S)-2-(3-(5-((1-Acryloylazetidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-(hydroxymethyl)phenyl)-6-cyclopropyl- 3,4-dihydroisoquinolin-1(2H)-one; (R)-N-(3-(5-((1-Acryloylazetidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2 -fluorobenzamide; (R)-N-(3-(5-((1-acryloylpiperidin-3-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4-cyclopropyl-2-fluorobenzamide ; N-(3-(5-(((2R,3S)-1-acryloyl-3-methoxypyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4 -cyclopropyl-2-fluorobenzamide; N-(3-(5-(((2S,4R)-1-acryloyl-4-cyanopyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)-4 -cyclopropyl-2-fluorobenzamide; or N-(3-(5-(((2S,4S)-1-acryloyl-4-cyanopyrrolidin-2-yl)methoxy)-6-aminopyrimidin-4-yl)-5-fluoro-2-methylphenyl)- 4-cyclopropyl-2-fluorobenzamide.

Unless otherwise indicated, the chemical terms used above in describing the BTK inhibitor of Formula I are used according to their meanings given in international application WO/2015/079417, which is hereby incorporated by reference in its entirety.

Examples of JAK inhibitors

In one embodiment, the combination described in this application includes a Janus kinase (JAK) inhibitor. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., colon cancer, prostate cancer, lung cancer, breast cancer, or pancreatic cancer), e.g., hemoblastosis ( for example, leukemia (eg, myeloid leukemia or lymphocytic leukemia), eg, lymphoma (eg, non-Hodgkin's lymphoma), or multiple myeloma.

In some embodiments, the JAK inhibitor is 2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide ( Compound A17) or its dihydrochloride, or the compound disclosed in PCT publication WO 2007/070514.

In some embodiments, a JAK inhibitor, e.g., 2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl) benzamide (Compound A17) or its dihydrochloride, or a compound disclosed in PCT publication WO 2007/070514, is administered at a dose of about 400-600 mg (e.g. per day), such as about 400, 500 or 600 mg or about 400-500 or 500-600 mg.

In some embodiment, the JAK inhibitor is ruxolitinib phosphate (also known as Jakavi®; Compound A18) or a compound disclosed in PCT publication WO 2007/070514.

In one embodiment, a JAK inhibitor, for example, ruxolitinib phosphate (also known as Jakavi; Compound A18) or a compound disclosed in PCT publication WO 2007/070514, is administered at a dose of about 15-25 mg, for example, twice a day. In some embodiments, the dose is about 15, 20, or 25 mg, or about 15-20 or 20-25 mg.

Examples of CRTH2 inhibitors

In one embodiment, the combination described herein comprises an inhibitor of the chemoattractant receptor homologous to T-helper cell 2 (CRTH2). In some embodiments, the combination is used to treat cancer, such as the cancer described herein.

In some embodiments, the CRTH2 inhibitor is QAV680 (CAS Number: 872365-16-7). QAV680 is also known as fevipiprant and 2-[2-methyl-1-[(4-methylsulfonylphenyl)-methyl]pyrrolo[2,3-b]pyridin-3-yl]acetic acid. QAV680 is disclosed, for example, in Sandham et al. Bioorg Med Chem. 2013; 21(21):6582-91. QAW039 is also known as [1-(4-methanesulfonyl-2-trifluoromethyl-benzyl)-2-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl]acetic acid. QAW039 is disclosed, for example, in Sykes et al. European Respiratory Journal September 1, 2014 vol. 44 no. Suppl 58 P4074.

In some embodiments, the CRTH2 inhibitor is QAW039 (CAS Number: 872365-14-5).

Other CRTH2 inhibitors that may be used in combination include, for example, AZD1981, ARRY-502, setipiprant (ACT-453859) and ACT-129968.

Examples of PFKFB3 inhibitors

In one embodiment, the combination described herein comprises a 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) inhibitor. In some embodiments, the combination is used to treat cancer, e.g., the cancer described herein, e.g., a solid tumor (e.g., an advanced solid tumor), e.g., ovarian cancer, fallopian tube cancer, colon and rectal cancer (CRC), leukemia , lung cancer, prostate cancer, breast cancer, or pancreatic cancer. In some embodiments, the cancer is ovarian cancer (eg, recurrent ovarian cancer), fallopian tube cancer, or colon and rectal cancer (eg, metastatic colon and rectal cancer). Without being limited by theory, it is believed that in some embodiments, PFKFB3-directed glycolysis can regulate vascular branching in a cancerous tumor, wherein endothelial cells can use glycolysis for ATP synthesis and metastatic potential. In some embodiments, the combination is used to treat cancer with high angiogenic activity. In other embodiments, the combination is used to treat cancer enriched in Th17 cells. In other embodiments, the combination is used to treat cancer that does not respond or moderately responds to anti-PD-1 or anti-PD-L1 antibody monotherapy.

In some embodiments, the PFKFB3 inhibitor is PFK-158. PFK-158 is also known as ACT-PFK-158 or (E)-1-(pyridin-4-yl)-3-(7-(trifluoromethyl)quinolin-2-yl)-prop-2-en-1-one (Accident Number 1462249-75-7). PFK-158 is a derivative of 3-(3-pyridinyl)-1-[4-pyridinyl]-2-propen-1-one (3PO). PFKFB3, which catalyzes the conversion of fructose-6-phosphate to fructose-2,6-bisphosphate, is highly expressed and active in human cancer cells and also plays an important role in increasing glycolytic flux and cancer cell proliferation. In some embodiments, PFKFB3 is expressed (e.g., overexpressed over normal tissue) in a cancer selected from leukemia, colon cancer, lung cancer, prostate cancer, ovarian cancer, fallopian tube cancer, breast cancer, or pancreatic cancer. PFKFB3 inhibitors, such as PFK-158, can bind to and inhibit PFKFB3 activity, resulting in inhibition of the glycolytic pathway and glucose uptake by cancer cells. This prevents macromolecule synthesis and energy production, which cause increased cell proliferation in cancer cells compared to normal, healthy cells. Depriving cancer cells of nutrients and energy results in inhibition of cancer cell growth. In some embodiments, a PFKFB3 inhibitor, such as PFK-158, inhibits glucose uptake by the cancer cell. In other embodiments, a PFKFB3 inhibitor, such as PFK-158, reduces pathological angiogenesis. For example, PFK-158 can reduce 18FDG uptake in tumor xenografts and exhibits antitumor activity in mice with low white blood cell count (LLC). PFK-158 is disclosed, for example, on page 5 of WO 2013/148228.

In some embodiments, the PFKFB3 inhibitor has the following structure:

.

.

In some embodiments, the implementation of the combination described in this application is used to treat ovarian cancer. Ovarian cancer is commonly known as an immunoreactive disease (Zhang et al., N Engl J Med. 2003; 348(3):203-13). For example, an improvement in overall survival was seen in patients with higher intraepithelial CD8+ T cell counts compared with patients with lower CD8+ T cell counts (median survival 55 months vs 26 months; hazard ratio=0.33; P<0.001) (Sato et al. Proc Natl Acad Sci U S A. 2005; 102(51):18538-43). In some embodiments, increased Tregs can be used as indicators of poor survival (Curiel et al. Nat Med. 2004; 10(9):942-9). Without being limited by theory, it is believed that in some embodiments, a high tumor antigen load may lead to a progressive loss of one or more antigen-specific T cell effector functions, for example, leading to T cell depletion. Patients with ovarian cancer, such as advanced ovarian cancer, may be selected for immunotherapy, such as the combination therapy described herein, at least in part because of the low burden of disease after primary therapy, as well as the high risk of relapse.

In some embodiments, the combination further comprises an IDO inhibitor (eg, an IDO inhibitor as described herein). In other embodiments, the combination further comprises a CD-27 agonist (eg, a CD-27 agonist described herein). In other embodiments, the combination further comprises an anti-CTLA-4 antibody (eg, an anti-CTLA-4 antibody described herein, eg, ipilimumab). In other embodiments, the combination further comprises a PARP inhibitor (eg, the PARP described herein, eg, ipilimumab). In other embodiments, the combination further comprises chemotherapy (eg, the chemotherapy described herein, eg, paclitaxel). In other embodiments, the combination further comprises a CSF-1R inhibitor (eg, a CSF-1R inhibitor as described herein).

In some embodiments, the implementation of the combination described in this application is used to treat cancer of the colon and rectum. Colon and rectal cancer (CRC) is usually considered an inflammatory cancer. Metastatic, or stage IV, colon cancers have a 5-year relative survival rate of approximately 11%. Without being limited by theory, it is believed that in some embodiments, the development of CRC in humans is associated with the interleukin-23 (IL-23)-IL-17 pathway (Grivennikov et al., Nature. 2012; 491(7423):254-8). IL-17 is associated with poor prognosis and induces angiogenesis by stimulating VEGF production of cancer cells in CRC. Activated gdT17 cells also secrete other cytokines, including IL-8, TNF-alpha, and GM-CSF, which induce chemoattraction of MSCs into the tumor and maintain their immunosuppressive activity. Th17 cell expression profiles in CRC have been shown to be associated with poor survival (Grivennikov et al., Nature. 2012; 491(7423):254-8; Zou and Restifo, Nat Rev Immunol. 2010; 10(4):248 -56). CRC is often sensitive to VEGF inhibition, however, an IL-17-mediated paracrine network has been shown to promote tumor resistance to antiangiogenic therapy (Chung et al. Nat Med. 2013; 19(9):1114-23). IL-17-induced recruitment of CD11b+Gr1+ myeloid cells into the tumor microenvironment can induce VEGF-independent tumor growth.

In some embodiments, the combination is used to treat a subject having microsatellite stable (MSS) colorectal cancer. In some embodiments, the subject does not respond or responds poorly to checkpoint inhibitor monotherapy. In other embodiments, the combination is used to treat a subject having microsatellite unstable colon and rectal cancer (MSI). In some embodiments, the subject has MSI colon and rectal cancer or stage IV colon and rectal cancer. In some embodiments, the subject is responsive to checkpoint inhibitor therapy.

In some embodiments, the implementation of the combination further includes chemotherapy, for example, the chemotherapy described in this application. In some embodiments, the combination further comprises 5-FU, folinic acid, or both. Alternatively, or in combination, the combination further comprises oxaliplatin. Alternatively, or in combination, in other embodiments, the combination further comprises irinotecan. In some embodiments, the combination further comprises FOLFOX or FOLFIRI. In some embodiments, the combination further comprises an angiogenesis inhibitor (eg, bevacizumab).

In some embodiments, a PFKFB3 inhibitor, PFK-158, or a compound disclosed in PCT publication WO 2013/148228 is used in combination with a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule). In some embodiments, the PFKFB3 inhibitor, PFK-158, or a compound disclosed in PCT publication WO 2013/148228 is administered intravenously. In some embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg), e.g., once every 4 weeks, e.g., by intravenous infusion. In other embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 400 mg (e.g., 300 mg), e.g., once every 3 weeks, e.g., by intravenous infusion. In some embodiments, the combination is used to treat the cancer described herein, eg, ovarian cancer, fallopian tube cancer (eg, papillary serous adenocarcinoma), or colon and rectal cancer. In some embodiments, the combination is used to treat a subject who has responded poorly or has not responded to chemotherapy (eg, platinum-derived chemotherapy or 5-FU chemotherapy).

Examples of chemotherapy

In one embodiment, the combination described in this application includes a chemotherapeutic agent. In some embodiments, the combination is used to treat cancer, eg, the cancer described herein, eg, breast cancer, lung cancer (eg, non-small cell lung cancer (NSCLC)), nasopharyngeal cancer, or pancreatic cancer. In some embodiments, the cancer is breast cancer, eg, metastatic breast cancer, eg, HER2-negative breast cancer (eg, triple-negative breast cancer (TNBC) or HR+ endocrine therapy-resistant breast cancer). In some embodiments, the lung cancer (eg, non-small cell lung cancer) is squamous NSCLC or non-squamous NSCLC. In some embodiments, the combination is used to treat cancer of the nasopharynx.

In some embodiments, the chemotherapeutic agent is paclitaxel, for example, albumin nanoparticle-bound paclitaxel, also known as albumin-bound paclitaxel or paclitaxel-albumin (Abraxane®). In some embodiments, paclitaxel has the following structure:

Paclitaxel is also known as (2α,4α,5β,7β,10β,13α)-4,10-bis(acetyloxy)-13-{[(2R,3S)-3-(benzoylamino)-2-hydroxy-3-phenylpropanoyl ]oxy}-1,7-dihydroxy-9-oxo-5,20-epoxytax-11-en-2-yl benzoate.

Paclitaxel-albumin is a paclitaxel bound to nanoparticles of albumin, which is different from conventional paclitaxel, which includes Kolliphor EL (formerly Cremophor EL) in the dosage form. This solvent-free formulation was designed to improve the therapeutic index of paclitaxel, especially to reduce toxic events such as allergic reactions (thus avoiding premedication with antihistamines and steroids), with the potential to increase the uptake of paclitaxel in cancer cells (Ibrahim et al. Clin Cancer Res. 2002;8(5):1038-44). The solvent used for conventional paclitaxel can also interfere with drug delivery to the tumor, limiting its clinical efficacy (ten Tije et al. Clin Pharmacokinet. drugs in a narrow dose range, which is most likely due to the capture of paclitaxel in solvent micelles (Gianni et al. J Clin Oncol. 1995; 13(1):180-90; van Tellingen et al. Br J Cancer. 1999; 81( 2):330-5; Sparreboom et al. Cancer Res. 1999; 59(7):1454-7). Compared to conventional paclitaxel, paclitaxel-albumin creates a 10-fold higher mean maximum concentration of free paclitaxel, and in addition, paclitaxel-albumin passes through endothelial cell layers faster and shows higher tissue penetration and slower elimination of paclitaxel. According to preclinical models, increased intratumoral delivery and retention results in 33% higher drug concentrations in the tumor (Desai et al. Clin Cancer Res. 2006; 12(4):1317-24. Error corrected in: Clin Cancer Res. 2006;12(12):3869).

Without being limited by theory, it is believed that in some embodiments, the implementation of paclitaxel-albumin is a microtubule inhibitor that causes the assembly of microtubules from tubulin dimers and stabilizes the microtubules by blocking depolymerization. This stability leads to inhibition of the normal dynamic reorganization of the microtubule network, which is essential for vital interphase and mitotic cellular functions.

In some embodiments, paclitaxel-albumin can be used as monotherapy, with a recommended dose of 260 mg/m ² every 3 weeks, for the treatment of patients with metastatic breast cancer, for example, after first-line chemotherapy has failed due to metastasis or relapse within 6 months of adjuvant chemotherapy (previous therapy should have included an anthracycline unless clinically contraindicated). Phase III results demonstrated significantly improved overall patient response rate (ORR) (33% vs. 19%; P=0.001) and time to disease progression (TTP) (23.0 vs. 16.9 weeks; HR=0.75; P =0.006) in patients receiving first and second line or subsequent paclitaxel-albumin versus patients receiving conventional paclitaxel, 175 mg/m ² every 3 weeks (Gradishar et al. J Clin Oncol. 2005; 23(31 ):7794-803).

In some embodiments, a weekly (Day 1, 8, 15 every 4 weeks) paclitaxel-albumin regimen may be used. Two different weekly doses (100 mg/m ² and 150 mg/m ² qw3/4) were compared in a randomized phase II trial with 300 mg/m ² paclitaxel-albumin every 3 weeks (q3w) and 100 mg/m ² docetaxel q3w in the setting of first-line therapy (Gradishar et al. J Clin Oncol. 2009; 27(22):3611-9. Erratum in: J Clin Oncol. 2011; 29(19):2739; Gradishar et al. Clin Breast Cancer. 2012;12(5):313-21). In some embodiments, the implementation of the dose of 150 mg/m ² qw3/4 is used, for example, for patients who need a rapid effect of anticancer therapy. In other embodiments, a dose of 100 mg/m ² qw3/4 is used, for example, due to a more controlled safety profile.

The pharmacokinetics of the total paclitaxel after 30 and 180-minute infusions of paclitaxel-albumin at doses of 80-375 mg/m ² was determined in clinical studies. Dose levels in mg/m ² refer to mg of paclitaxel in paclitaxel-albumin. Following intravenous administration of paclitaxel-albumin, plasma concentrations of paclitaxel decreased in a two-phase process, with an initial rapid distribution to the peripheral compartment and a slower second phase representing drug elimination. The drug exposure (AUC) was dose-dependent in the range of 80-300 mg/m ² , while the pharmacokinetics of paclitaxel in the case of paclitaxel-albumin did not depend on the duration of intravenous administration. The average total clearance varied from 13 to 30 l/h/m ² and the average half-life in the final phase varied from 13 to 27 hours.

In some embodiments, the combination described herein is used to treat breast cancer, eg, metastatic breast cancer or HER2-negative breast cancer (eg, triple-negative breast cancer). Without being limited by theory, it is believed that in some embodiments, the use of steroids in combination with anti-PD-1 compounds can be antagonistic, with paclitaxel-albumin having an advantage over other taxanes, at least in part, since premedication with corticosteroids is not required.

In some embodiments, the combination is used to treat a subject having triple negative breast cancer or HR+ breast cancer after failure of endocrine therapy. In other embodiments, the combination is used to treat a taxan-naive patient or a patient having at least 1 year disease-free survival following their use in an adjuvant therapy regimen. In some embodiments, the combination is used to treat a subject who is eligible for first line chemotherapy.

In breast cancer, the interaction of the immune system with tumor cells and the tumor microenvironment may play an important role, for example, in the case of TNBC and HER2+ subtypes. Breast cancer in general, and TNBC subtypes in particular, often contains tumor infiltrating lymphocytes (TIL) in the stroma and is associated with high PD-L1 expression (up to 50% of tumors). Without being limited by theory, it is believed that in some embodiments, cytotoxic chemotherapeutic agents can increase immune priming, T cell migration, and/or immunoeffector phases of the antitumor immunity cycle, while increasing antigenicity, immunogenicity, and immune killing sensitivity of cancer cells, which increases the potential for response to checkpoint inhibitors.

In some embodiments, the combination is used to treat a subject who develops or is at risk of developing resistance to anti-PD-1 or anti-PD-L1 therapy. For example, preclinical and clinical results indicate that several cytotoxic and anticancer therapies may help overcome some of the potential mechanisms of resistance to PD-L1/PD-1 inhibition (e.g., a disturbance in the activation of cancer antigen-specific T cells or a lack of immunocyte infiltration into tumors) by increasing T cell migration and tumor bed infiltration (Zitvogel et al. 2013). Several cytotoxic agents (including paclitaxel) can also directly affect the phenotype and function of dendritic cells (DCs), causing them to mature and secrete pro-inflammatory cytokines and increase their antigen presentation function (eg, expression of MHC class II molecules on their surface), resulting in increased priming of CD8+ T cells and increased cytotoxic activity (Pfannenstiel et al. Cell Immunol. 2010; 263(1):79-87). It has also been suggested that chemotherapy could stimulate in tumor cells their antigenicity (causing somatic mutations and increasing the exposure of various molecules on the surface of tumor cells that act as new antigens), immunogenicity (through the secretion of apoptosis associated molecules (CDAM)) and sensitivity to immune attack (by expression of co-stimulatory molecules such as B7-1 on the surface of malignant cells or downregulation of immunosuppressive molecules such as PD-L1 and PD-L2) (Hodge et al. Int J Cancer. 2013; 133(3) :624-36). In preclinical in vivo models , platinum- or taxane-based chemotherapy demonstrates a synergistic effect in combination with anti-PD-L1 antibodies and induces long-term antitumor responses associated with increased numbers of tumor-infiltrating CD8+ T cells (Giaccone et al. 2015, Adams et al. 2015 , Liu et al. 2015). In clinical settings, increased PD-L1 expression and infiltration of CD8+ T cells into tumors were observed in patients undergoing serial biopsy in a phase 1b study testing the combination of a PD-L1 inhibitor (atezolizumab) and chemotherapy for NSCLC (Camidge et al. 2015). The development of new intratumoral immune cell infiltrates has also been observed following neoadjuvant therapy with paclitaxel in early stage breast cancer with clinical and laboratory responses directly correlated with immune cell infiltration into tumors (Demaria et al. Clin Cancer Res. 2001; 7( 10):3025-30).

In some embodiments, the combination is used to treat a subject who has exhibited immune infiltration, eg, containing tumor infiltrating lymphocytes (TILs), for cancer. In other embodiments, the subject is selected for treatment as a result, at least in part, of the presence of TIL lymphocytes in the cancer. For example, more than 70% of breast cancers contain tumor-infiltrating lymphocytes in the stroma, and preclinical data indicate that in most cases of breast cancer, the local immune microenvironment plays a role in controlling cancer progression. Several studies have shown that immune infiltration in breast cancer correlates with a better prognosis and better response to chemotherapy (Denkert et al. J Clin Oncol. 2010; 28(1):105-13; Loi et al. Oncoimmunology. 2013; 2(7):e24720). In the case of TNBC, approximately 15% of tumors show a high degree of lymphocytic infiltration at diagnosis, with the majority (65-80%) containing low or moderate levels of immune cells (Ruffell et al. Proc Natl Acad Sci U S A. 2012; 109(8): 2796-801 Garcia-Teijido et al Clin Med Insights Oncol 2016 10(Suppl 1):31-9). Analysis of gene expression profiling has also demonstrated an association between immunomodulatory gene expression and better treatment outcome in TNBC (Desmedt et al. J Clin Oncol. 2016; 34(16):1872-81). In 2 different studies, one in TNBC specimens and the other in all breast cancer types, PD-L1 expression was high (19% in TNBC specimens, and 44% in all breast cancer specimens) and associated with more intense CD8+ T- cells than PD-L1 tumors in TNBC samples (Mittendorf et al. Cancer Immunol Res. 2014; 2(4):361-70., Ghebeh et al. Neoplasia. 2006; 8(3):190-8). In addition, TNBC appears to exhibit genetic instability with a high mutation rate, which could also lead to an increase in immunogenic neoantigens and infiltration of antitumor T cells (Wang et al. Ann Oncol. 2015; 26(3):523-8) . In HER2 positive breast cancer, TIL and immune signatures are also associated with better prognosis (Salgado et al. JAMA Oncol. 2015; 1(4):448-54). HER2-positive breast cancer is also a highly proliferating subtype of breast cancer with an increased level of genomic instability. Overexpression of HER2 itself can act as a tumor-associated antigen, inducing the immune system. In the case of highly proliferative HR+ cancers (luminal B subtype), immune cells predict a better prognosis (Bianchini et al. J Clin Oncol. 2010; 28(28):4316-23. Erratum in: J Clin Oncol. 2010; 28(32 ):4868 J Clin Oncol 2012;30(6):679). In HR+ breast cancer, the interaction between tumor cells and the immune environment may be based on different mechanisms than in other breast cancer subtypes, including modulation of the tumor microenvironment by mutual influences of endocrine factors, pro-inflammatory status and immune cells (Dieci et al. Cancer Treat Rev. 2016 46:9-19).

In some embodiments, a chemotherapeutic agent, e.g., paclitaxel-albumin, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), are administered at a dose and/or timing that, in combination provide the desired antitumor activity.

In one embodiment, a chemotherapeutic agent, such as paclitaxel-albumin, is administered at a dose of 25 mg/m ² to 300 mg/m ² , such as 25 mg/m ² to 250 mg/m ² , 50 mg/m ² up to 200 mg/ ^m2 , from 100 mg/ ^m2 to 150 mg/ ^m2 , from 75 mg/ ^m2 to 125 mg/ ^m2 , from 125 mg/ ^m2 to 175 mg/ ^m2 , from 50 mg/m2 m ² to 100 mg / m ² , from 100 mg / m ² to 200 mg / m ² or from 200 mg / m ² to 250 mg / m ² , for example, at a dose of 50 mg / m ² , 75 mg / m ² , 100 mg/m ² , 125 mg/m ² , 150 mg/m ² , 175 mg/m ² , 200 mg/m ² , 220 mg/m ² or 260 mg/m ² , for example, three times (for example, weekly) for a period of four weeks, e.g. weekly on Day 1, Day 8 and Day 15 every four weeks (qw3/4), e.g. intravenously.

In one embodiment, a chemotherapeutic agent, such as paclitaxel-albumin, is administered at a dose of 50 mg/m ² to 150 mg/m ² (eg, 100 mg/m ² ), such as weekly on Day 1, Day 8, and Day 15 every four weeks (qw3/4). In some embodiments, a chemotherapeutic agent, such as paclitaxel-albumin, is administered intravenously. In one embodiment, a chemotherapeutic agent, such as paclitaxel-albumin, is administered at a dose of 50 mg/m ² to 150 mg/m ² (eg, 100 mg/m ² ), such as weekly on Day 1, Day 8, and Day 15 every four weeks (qw3/4), for example by intravenous infusion, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg), for example, once every 4 weeks or once every 8 weeks, for example by intravenous infusion. In some embodiments, the infusion of a chemotherapeutic agent, e.g., paclitaxel-albumin, is started after (e.g., at least 30 minutes, 45 minutes, 60 minutes, 75 minutes, 90 minutes, or 120 minutes after) completion of the PD-1 inhibitor infusion (e.g., , anti-PD-1 antibody molecules). For example, a chemotherapeutic agent, eg, paclitaxel-albumin and/or a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule), may be administered by 30-120 minute (eg, 30 minute) intravenous infusion. In an embodiment, the course of treatment is 28 days or 56 days (eg, 28 days).

In some embodiments, the chemotherapeutic agent includes cisplatin, pemetrexed, or both. Cisplatin is also known as cisplatin, platinum, neoplatin, cismaplat, or cis-diamminedichloroplatinum (II) (CDDP). Pemetrexed is also known as (S)-2-(4-(2-(2-amino-4-oxo-4,7-dihydro-3H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl)benzamido ) pentanedioic acid. In some embodiments, the combination is used to treat cancer, such as the cancer described herein. In some embodiments, the cancer is lung cancer, such as non-small cell lung cancer (NSCLC). In some embodiments, the NSCLC is non-squamous NSCLC. In some embodiments, the cancer is nasopharyngeal cancer.

In some embodiments, the chemotherapeutic agent includes paclitaxel, carboplatin, or both. Carboplatin is also known as cis-diammine(cyclobutane-1,1-dicarboxylate-O,O')platinum (II). In some embodiments, the combination is used to treat cancer, such as the cancer described herein. In some embodiments, the cancer is lung cancer, such as non-small cell lung cancer (NSCLC). In some embodiments, the NSCLC is squamous NSCLC. In some embodiments, the cancer is nasopharyngeal cancer.

Other examples of chemotherapeutic agents that can be used in combination include, but are not limited to, an alkylating agent (e.g., a bifunctional alkylating agent (e.g., cyclophosphamide, mechlorethamine, chlorambucil, or melphalan), a monofunctional alkylating agent (dacarbazine (DTIC), nitrosoureas, temozolomide ( oral dacarbazine)), anthracycline (eg, daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, or valrubicin), cytoskeletal disruptors (taxane) (eg, paclitaxel, docetaxel, abraxane, or taxotere), epothilone, histone deacetylase inhibitor (eg, vorinostat, or romidepsin), a topoisomerase I inhibitor (eg, irinotecan or topotecan), a topoisomerase II inhibitor (eg, etoposide, teniposide, or tafluposide), or kinase inhibitors (eg, bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, vismodegib), a nucleotide or precursor analog ( e.g. azacitidine, imuran, capecitabine, cytarabine , doxyfluridine, fluorouracil, gemcitabine, hydroxyurea, mercaptopurine, methotrexate, thioguanine), peptide antibiotic (eg, bleomycin or actinomycin), platinum-based agent (eg, carboplatin, cisplatin, or oxaliplatin), retinoid (eg, tretinoin, alitretinoin, bexarotene) or a vinca alkaloid or derivative (eg vinblastine, vincristine, vindesine or vinorelbine).

In some embodiments, the combination further comprises an angiogenesis inhibitor, an EGFR inhibitor, a PARP inhibitor (e.g., BSI-201), an antibody-drug conjugate (e.g., GPNMB-specific (e.g., glembatumumab vedotin (CDX-011)) -132)), a Frizzled receptor inhibitor (eg, vantictumab), a PD-L1 inhibitor (eg, atezolizumab), surgery, radiation therapy, or a combination thereof.

In one embodiment, the combination described herein comprises a chemotherapeutic agent (eg, a hypomethylating agent, eg, decitabine). In one embodiment, the combination described herein further comprises an immune checkpoint molecule inhibitor, e.g. a PD-1 inhibitor (e.g. an anti-PD-1 antibody molecule described herein), a TIM-3 inhibitor (e.g. an antibody molecule against TIM-3 described in this application) or both. In some embodiments, the combination is used to treat cancer, eg, the cancer described herein, eg, hemoblastosis, eg, leukemia (eg, acute myeloid leukemia (AML)), or myelodysplastic syndrome (MDS). In some embodiments, the cancer is leukemia, eg, relapsed or refractory AML, or de novo AML (eg, substandard AML). In other embodiments, the cancer is MDS, eg high risk MDS.

In some embodiments, the chemotherapeutic agent is a hypomethylating agent. A hypomethylating agent (or demethylating agent) is an agent that inhibits DNA methylation. In some embodiments, a hypomethylating agent blocks the activity of a DNA methyltransferase (ie, a DNA methyltransferase inhibitor/DNMT inhibitor). Examples of hypomethylating agents that can be used in the combination described herein include decitabine and azacitidine.

In some embodiments, the chemotherapeutic agent is decitabine. Decitabine (also known as 5-aza-2'-deoxycytidine or Dacogen®) is a cytidine deoxynucleoside analogue. It can selectively inhibit DNA methyltransferase at low doses. Inhibition of DNA methyltransferase by decitabine can lead to hypomethylation of gene promoters, which can lead to reactivation of tumor suppressor genes or induction of cell differentiation or cellular senescence followed by apoptosis. Decitabine is indicated for the treatment of adults aged 65 years and older with newly diagnosed de novo or secondary WHO AML who are not eligible for standard induction chemotherapy. The use of decitabine was studied in patients with newly diagnosed de novo or secondary AML according to the WHO classification (Malik Malik and Cashen. Cancer Manag Res. 2014; 6: 53-61). The median overall survival (OS) for all treated patients was 7.7 months in patients treated with decitabine. The percentage of patients in complete remission (CR+ complete remission with incomplete platelet recovery (CRp)) was 17.8% (Lubbert et al. J Clin Oncol. 2011; 29(15):1987-96). Decitabine is indicated for the treatment of patients with MDS, such as previously treated and untreated, de novo and secondary MDS of all subtypes of the Franco-American-British classification, as well as intermediate-1, intermediate-2 and high risk groups according to the International Predictive Scoring System , in the USA.

In some embodiments, the combination is used to treat acute myeloid leukemia (AML). AML is a clonal neoplasia of the hematopoietic system, which is characterized, for example, by the proliferation of abnormal myeloid blast cells in the bone marrow. The proliferation of abnormal myeloid blast cells can disrupt normal hematopoiesis, resulting in the accumulation of dysfunctional, immature myeloid cells in the bone marrow and peripheral blood. In some embodiments, the diagnosis of AML is made when myeloid blast cells comprise ≥20% of nucleated cells in a bone marrow aspirate. These leukemic blast cells can inhibit normal hematopoietic stem and progenitor cells and influence the appearance of mature, differentiated hematopoietic cells in the peripheral blood. Clinical symptoms may include, for example, anemia, neutropenia, or thrombocytopenia, which can lead to life-threatening infection and bleeding. AML may occur in the setting of prior myelodysplastic syndrome ("secondary AML") or prior cytotoxic chemotherapy ("treatment-related AML").

Long-term disease control can be achieved in patients with good prognostic parameters, such as younger age and favorable cytogenetics. Patients under 60 without significant comorbidities may initially be treated with intensive myeloablative induction chemotherapy, which typically includes anthracycline and cytarabine. The goal of this therapy is to achieve, for example, complete remission (CR) with less than 5% blast cells under conditions of normal hematopoiesis, which can be achieved in more than 65% of cases in this group of patients. Post-remission consolidation therapy using chemotherapy or either autologous or allogeneic hematopoietic stem cell transplantation may subsequently be used to increase the duration of remission. There are relatively high recurrence rates of AML for which limited effective treatment options are available. Elderly patients usually suffer even worse, achieving significantly lower and shorter response times and experiencing more treatment complications. For this group of elderly patients who cannot tolerate intensive care, options are extremely limited and no effective standard approach exists. Treatment options include, for example, low dose cytarabine or hypomethylating agents such as 5-azacytidine or decitabine. There is a need for more effective treatments to improve the survival of patients with AML, for example, elderly patients with AML and patients with relapsed or refractory disease.

Myelodysplastic syndrome (MDS) is a heterogeneous group of chronic myeloid disorders that include persistent peripheral blood cytopenias and an increased incidence of leukemic transformation. Clinical findings are variable and the diagnosis is usually made on the basis of morphological and cytogenetic abnormalities of hematopoietic cells in the bone marrow and peripheral blood. MDS prognosis can be determined according to the revised International Prognostic Scoring System (rIPSS), which is based on clinical cytogenetics, bone marrow blast percentages, and cytopenia. Patients are classified into five risk categories according to total scores based on these prognostic factors (Greenberg et al. J Natl Compr Canc Netw. 2013; 11(7):838-74). Patients diagnosed with high and very high risk MDS according to rIPSS have a high percentage of transformation into AML and a poor prognosis. Hypomethylating agents are considered the standard of care for MDS (NCCN Guidelines for Patients®, Myelodysplastic Syndromes, Version 1.2016, National Comprehensive Cancer Network).

Without being limited by theory, it is believed that in some embodiments, antitumor immunity can be used to treat AML and improve patient outcomes, for example, based on the antileukemic activity seen with allogeneic hematopoietic stem cell transplantation and donor lymphocyte infusions. There is evidence of a suppressive immune microenvironment in AML with overexpression of PD-L1 in leukemic blast cells. Preclinical data in a mouse model of AML indicate that disease progression is associated with increased expression of PD-1 on CD8-positive T cells, and that treatment with anti-PD-1 or anti-PD-L1 antibodies reduces the burden of AML and improves survival in these models of this study. diseases. Patients have evidence of elevated PD-L1 expression in a subgroup of AML/MDS patients that continues to increase with progression.

In addition to activating the PD-1/PD-L1 pathway, there is evidence that AML stem cells co-express the TIM-3 immune checkpoint. TIM-3 can, together with the PD-1/PD-L1 pathway, suppress tumor recognition by cytotoxic T cells. TIM-3 is a unique AML stem cell antigen that is present in leukemia blast cells but absent in normal hematopoietic stem cells, and anti-TIM3 antibody treatment has been shown to be effective in reducing leukemia burden in a mouse model of AML (Kikushige et al. Cell Stem Cell 2010;7(6):708-17). There is also evidence in MDS that treatment with the standard of care hypomethylating agent, decitabine, increases PD-1 expression in T cells, highlighting its role in immunomodulation.

Without being limited by theory, it is believed that in some embodiments, chemotherapeutic agents (e.g., hypomethylating agents, e.g., decitabine) may cause an increase in the expression of immune checkpoints (e.g., PD-1 and/or TIM-3) that may confer immune escape and lead to therapeutic resistance. Without being limited by theory, it is also believed that in some embodiments, AML and MDS have increased co-expression of the immune checkpoint receptors PD-1 and TIM-3, which co-inhibit immune recognition by cytotoxic T cells. Preclinical data indicate that concurrent blockade of PD-1 and TIM-3 elicits greater T cell activation than single therapy (Sakuishi et al. J Exp Med. 2010; 207(10):2187-94, Ngiow et al. Cancer Res. 2011; 71(10):3540-51), and synergistically inhibits tumor growth in experimental cancer models (Anderson et al. Cancer Immunol Res. 2014; 2(5):393-8, Ngiow et al. Cancer Res 2011;71(10):3540-51). In some embodiments, the combination described herein optimizes the approach to checkpoint inhibition in AML and MDS, for example, by providing treatment-induced immunomodulation and co-inhibitory pathways.

In other embodiments, the combination comprises an inhibitor of an immune checkpoint molecule, eg, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein), and a chemotherapeutic agent (eg, a hypomethylating agent, eg, decitabine). In some embodiments, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered at a dose of 200 mg to 600 mg (e.g., 300 mg to 500 mg, e.g., 400 mg), for example, once every 4 weeks, for example, intravenously. In some embodiments, the chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered at a dose of 5 mg/m ² to 50 mg/m ² (e.g., 10 mg/m ² to 30 mg/m ² , e.g., in dose of 20 mg/m ² ), for example, daily, for example, for 1, 2, 3, 4, 5, 6, 7, 8 or more days, for example, intravenously. In an embodiment, an immune checkpoint molecule inhibitor, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), is administered on day 8 of a 28 day course, and a chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered on days 1, 2, 3, 4 and 5 of a 28 day course.

In other embodiments, the combination comprises an inhibitor of an immune checkpoint molecule, e.g., a TIM-3 inhibitor or an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274, and a chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) . In some embodiments, an immune checkpoint molecule inhibitor, such as a TIM-3 inhibitor or an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274, is administered at a dose of 10 mg to 1000 mg (e.g., 20 mg up to 800 mg, from 80 mg to 400 mg, for example, in a dose of 240 mg), for example, once every 2 weeks, for example, intravenously. In some embodiments, the chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered at a dose of 5 mg/m ² to 50 mg/m ² (e.g., 10 mg/m ² to 30 mg/m ² (e.g., in dose of 20 mg/m ² ), for example, daily, for example, for 1, 2, 3, 4, 5, 6, 7, 8 or more days, for example, intravenously. In an embodiment, an immune checkpoint molecule inhibitor, for example, a TIM-3 inhibitor or anti-TIM-3 antibody molecule disclosed in U.S. Patent Application Publication 2015/0218274 is administered on days 8 and 22 of a 28-day course, and a chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered on days 1, 2, 3, 4 and 5 of the 28-day course.

In other embodiments, the combination comprises a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein), a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274 ) and a chemotherapeutic agent (eg, a hypomethylating agent, eg, decitabine). In some embodiments, the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 600 mg (eg, 400 mg dose), eg, once every 4 weeks, eg, intravenously. In some embodiments, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) is administered at a dose of 10 mg to 800 mg (e.g., 20 mg to 400 mg, from 40 mg to 200 mg or 50 mg to 100 mg, eg in a dose of 80 mg), eg once every 2 weeks, eg intravenously. In some embodiments, the chemotherapeutic agent (e.g., a hypomethylating agent, e.g., decitabine) is administered at a dose of 5 mg/m ² to 50 mg/m ² (e.g., 10 mg/m ² to 30 mg/m ² (e.g., in dose of 20 mg/m ² ), for example, daily, for example, for 1, 2, 3, 4, 5, 6, 7, 8 or more days, for example, intravenously. In an embodiment, a PD-1 inhibitor (for example, a molecule anti-PD-1 antibody) is administered on day 8 of the 28 day course, a TIM-3 inhibitor (e.g., the anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) is administered on days 8 and 22 of the 28 day course. course and a chemotherapeutic agent (eg, a hypomethylating agent, eg, decitabine) is administered on days 1, 2, 3, 4, and 5 of a 28-day course.

In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274), injected on the same day. In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274), administered on different days. In some embodiments, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in U.S. Patent Application Publication 2015/0218274) is administered, for example, on the same day prior to administration of the PD-1 inhibitor (e.g., an antibody molecule against PD-1 described in this application). In other embodiments, a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in U.S. Patent Application Publication 2015/0218274) is administered, for example, on the same day after administration of the PD-1 inhibitor (e.g., an antibody molecule against PD-1 described in this application). In some embodiments, an infusion of saline or dextrose solution, e.g., over about 1 hour, may be performed between administration of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein) and administration of a TIM-3 inhibitor (e.g., , an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274). In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule described herein), a TIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274), or both are administered by intravenous infusion over 30 minutes to 2 hours, eg about 1 hour. A PD-1 inhibitor (eg, an anti-PD-1 antibody molecule described herein) and a TIM-3 inhibitor (eg, an anti-TIM-3 antibody molecule disclosed in US Patent Application Publication 2015/0218274) can be stored in the form liquid or lyophilisate. In some embodiments, the PD-1 inhibitor (e.g., the anti-PD-1 antibody molecule described herein) is stored in the form of a lyophilisate and the TIM-3 inhibitor (e.g., the anti-TIM-3 antibody molecule disclosed in U.S. Patent Application Publication 2015/0218274) is stored as a liquid.

Combination comprising a chemotherapeutic agent (for example, a hypomethylating agent (also sometimes known as a demethylating agent), for example, decitabine) and one or both of a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule described in this application) or a TIM inhibitor -3 (eg, anti-TIM-3 antibody molecules disclosed in US Patent Application Publication 2015/0218274) can be used in combination with an adjunctive therapy, eg for cancer treatment, eg anti-cancer therapy described herein. Examples of adjunctive therapies include, but are not limited to, arsenic trioxide (trisenox), all-trans retinoic acid (ATRA), stem cell or bone marrow transplant, a second chemotherapy agent (eg, azacitidine, cytarabine, daunorubicin, idarubicin, or lenalidomide), or radiation therapy. .

Examples of BRAF and MEK inhibitors

In one embodiment, the combination described in this application includes a BRAF inhibitor. In some embodiments, the combination is used to treat cancer, eg, the cancer described herein, eg, skin cancer (eg, melanoma). Without being limited by theory, it is believed that in some embodiments, the mitogen-activated protein kinase (MAPK) pathway is aberrantly activated in various forms of human cancer, for example, as a result of a BRAF kinase mutation that has been found in up to 50% of metastatic melanomas. In some embodiments, the combination is used to treat cancer having a BRAF mutation, such as the BRAF mutation described herein. In some embodiments, the combination is used to treat melanoma, such as unresectable or metastatic melanoma. In some embodiments, the combination is used to treat melanoma in a subject where the melanoma has a BRAF mutation (eg, a BRAF V600 mutation) and the subject has an elevated lactate dehydrogenase (LDH) level compared to a reference LDH level.

In some embodiments, the BRAF inhibitor is dabrafenib. Dabrafenib is also known as GSK2118436, N-{3-[5-(2-aminopyrimidin-4-yl)-2-tert-butyl-1,3-thiazol-4-yl]-2-fluorophenyl}-2,6- difluorobenzenesulfonamide or Tafinlar® (CAS No. 1195765-45-7). Dabrafenib is an orally bioavailable, potent and selective RAF kinase inhibitor of the human wild-type BRAF and CRAF enzymes, as well as mutant forms of the BRAF enzyme, such as BRAF V600E, BRAF V600K, and BRAF V600D. The mechanism of action of dabrafenib is consistent with competitive inhibition of ATP binding. In some embodiments, the combination is used to treat a subject who has been diagnosed with a cancer (eg, melanoma) having a BRAF mutation, eg, a BRAF mutation as described herein (eg, a BRAF V600 mutation). In other embodiments, the combination is used to treat cancer (eg, melanoma) other than wild-type BRAF cancer (eg, wild-type BRAF melanoma).

In some embodiments, the BRAF inhibitor or dabrafenib is administered at a dose of 50 mg to 300 mg (eg, 100 mg to 200 mg), eg, twice daily. In some embodiments, the BRAF inhibitor or dabrafenib is administered at a dose of 100 mg to 200 mg (eg, at a dose of about 150 mg), eg, twice daily. For example, a second dose of a BRAF inhibitor or dabrafenib may be administered approximately 12 hours after the first dose. In some embodiments, the BRAF inhibitor or dabrafenib is administered in a total daily dose of 100 mg to 600 mg (eg, 200 mg to 400 mg). In some embodiments, the BRAF inhibitor or dabrafenib is administered in a total daily dose of 200 mg to 400 mg (eg, a total daily dose of about 300 mg). In some embodiments, the BRAF inhibitor dabrafenib is administered orally.

In one embodiment, the combination described in this application includes a MEK inhibitor. In some embodiments, the combination is used to treat cancer, eg, the cancer described herein, eg, skin cancer (eg, melanoma). Without being limited by theory, it is believed that by inhibiting MEK, a member of the MAPK signaling cascade, cell proliferation can be blocked and apoptosis induced. In some embodiments, the combination is used to treat melanoma, such as unresectable or metastatic melanoma. In some embodiments, the combination is used to treat melanoma in a subject where the melanoma has a BRAF mutation (eg, a BRAF V600 mutation) and the subject has an elevated lactate dehydrogenase (LDH) level compared to a reference LDH level.

In some embodiments, the MEK inhibitor is trametinib. Trametinib is also known as GSK1120212, JTP-74057, TMT212, N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)-amino]-6,8-dimethyl-2,4,7- trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H)-yl}phenyl)acetamide or Mekinist® (CAS No. 871700-17-3). Without being limited by theory, it is believed that in some embodiments, trametinib is a reversible and highly selective allosteric inhibitor of MEK1 and MEK2. MEK proteins are important components of the MAPK pathway, which is normally overactivated in tumor cells such as melanoma cells. Oncogenic mutations in BRAF and RAS can signal through MEK1 or MEK2.

In some embodiments, the MEK inhibitor or trametinib is administered at a dose of 0.5 mg to 4 mg (eg, 1 mg to 3 mg), eg, once daily. In some embodiments, the MEK inhibitor or trametinib is administered at a dose of 1 mg to 3 mg (eg, a dose of about 2 mg), eg, once daily. In some embodiments, the MEK inhibitor or trametinib is administered orally.

In one embodiment, the combination described in this application includes a BRAF inhibitor and a MEK inhibitor.

Preclinical in vitro and in vivo data indicated enhanced antitumor activity of the combination of a BRAF inhibitor (eg, dabrafenib) and a MEK inhibitor (eg, trametinib). For example, the combination of dabrafenib and trametinib has shown increased antiproliferative activity against a group of BRAF mutant cell lines in vitro , suggesting a synergistic effect of dabrafenib and trametinib in addressing primary resistance to each individual agent. The combination was effective in inhibiting the growth of dabrafenib-resistant BRAF mutant melanoma cell clones, indicating the potential ability of the combination therapy to overcome acquired resistance. These cell line data are comparable to the in vitro results of other experimental combinations of BRAF and MEK inhibitors (Corcoran et al. Sci Signal. 2010; 3(149):ra84; Emery et al. Proc Natl Acad Sci US A. 2009; 106(48):20411-6).

As another example, the combination of dabrafenib and trametinib has shown increased activity in mouse xenograft models of BRAF mutant melanoma compared to either agent alone. In rat skin toxicity studies, the addition of trametinib to dabrafenib prevented the development of the proliferative skin lesions seen after treatment with dabrafenib alone. These results suggest that the addition of a MEK inhibitor to a BRAF inhibitor can suppress proliferative signals in normal skin cells that can lead to the development of hyperproliferative skin lesions, including, for example, keratoacanthoma and cutaneous squamous cell carcinomas often seen in clinical studies, including BRAF inhibitors (Flaherty et al. Curr Opin Oncol. 2010; 22(3):178-83; Chapman et al. Expert Opin Investig Drugs. 2011; 20(2):209-20; Robert et al. Curr Opin Oncol. 2011;23(2):177-82). Similar results were observed with another combination of BRAF and MEK inhibitors (Carnahan et al. Mol Cancer Ther. 2010; 9(8):2399-410).

In some embodiments, the BRAF inhibitor or dabrafenib is administered at a dose of 50 mg to 300 mg (e.g., 100 mg to 200 mg), e.g., twice daily, e.g., orally, and the MEK inhibitor or trametinib is administered at a dose of 0, 5 mg to 4 mg (eg, 1 mg to 3 mg), eg, once a day, eg, orally. In some embodiments, the BRAF inhibitor or dabrafenib is administered at a dose of 100 mg to 200 mg (e.g., about 150 mg), twice daily, orally, and the MEK inhibitor or trametinib is administered at a dose of 1 mg to 3 mg (e.g., , at a dose of approximately 2 mg), once a day, orally. For example, the first dose of the BRAF inhibitor or dabrafenib may be administered at the same time as the MEK inhibitor or trametinib, and the second dose of the BRAF inhibitor or dabrafenib may be administered approximately 12 hours after the first dose. In some embodiments, the BRAF inhibitor or dabrafenib is administered at a total daily dose of 100 mg to 600 mg (e.g., 200 mg to 400 mg), e.g., orally, and the MEK inhibitor or trametinib is administered at a dose of 0.5 mg to 4 mg (eg, 1 mg to 3 mg), eg, once a day, eg, orally. In some embodiments, the BRAF inhibitor or dabrafenib is administered in a total daily dose of 200 mg to 400 mg (e.g., in a total daily dose of approximately 300 mg), e.g., orally, and the MEK inhibitor or trametinib is administered in a dose of 1 mg to 3 mg ( eg at a dose of about 2 mg), eg once a day, eg orally.

In some embodiments, the combination comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule and: (i) a BRAF inhibitor or dabrafenib, (ii) a MEK inhibitor or trametinib, or (iii)-(i) and (ii). Without being bound by theory, it is believed that the use of an immune checkpoint molecule inhibitor, such as a PD-1 or PD-L1 inhibitor, in combination with a BRAF inhibitor (eg, dabrafenib), a MEK inhibitor (eg, trametinib), or both, may improve response (eg, , faster response, longer response, higher percentage response, or more complete response) caused by BRAF and/or MEK inhibition.

The following observations support, at least in part, a combinatorial or synergistic effect of targeted therapy (eg, BRAF-targeted therapy, MEK-targeted therapy, or both) with an immune checkpoint inhibitor (eg, a PD-1 inhibitor, such as an anti-PD- 1). For example, oncogenic BRAF can lead to immune escape of melanoma (Sumimoto et al. J Exp Med. 2006; 203(7):1651-6). Blocking BRAF activity results in increased expression of melanoma differentiation antigens in vitro (Kono et al. Mol Cancer Res. 2006; 4(10):779-92), e.g., through release of transcriptional repression and subsequent expression of MITF targets, e.g., MART-1 , gp100, TRP-1 and TRP-2 (Boni et al. Cancer Res. 2010; 70(13):5213-9). The effect of MAPK pathway inhibition on T cell activation and signaling has also been investigated in vitro . Although BRAF inhibitors have not shown adverse effects on T cell function and may even enhance their function through paradoxical signaling via RAS-GTP (Callahan et al. Cancer Immunol Res. 2014; 2(1):70-9), MEK inhibitors have shown dose-dependent inhibition of T cell function in vitro (Boni et al. Cancer Res. 2010; 70(13):5213-9; Vella et al. Cancer Immunol Res. 2014; 2(4):351-60). The in vivo antitumor activity of trametinib was also investigated in the syngeneic CT26 tumor model in immunocompetent BALB/C mice. In this study, trametinib monotherapy increased tumor-infiltrating CD4+ lymphocytes (TIL) and had no negative effect on the spread of CD8+ TIL. The combination of trametinib with an anti-mouse PD-1 antibody resulted in an effective antitumor response in a syngeneic mouse model of KRAS mutant colorectal tumor CT26 (Liu et al. Clin Cancer Res. 2015; 21(7):1639-51).

As another example, immune biomarker analysis was performed on tumor samples from 16 patients with metastatic melanoma treated with vemurafenib or dabrafenib plus trametinib. In this assay, BRAF inhibition was associated with increased expression of melanoma antigens (eg, MART, TYRP-1, TYRP-2, gp100), significantly elevated CD8+ TILs, decreased expression of immunosuppressive cytokines in the tumor microenvironment, and increased markers of T-cell toxicity, but also increased a marker of T cell depletion (eg, TIM-3, PD-1). It was also noted that antigen expression and CD8+ TIL were downregulated during melanoma progression but could be restored by co-inhibition of BRAF and MEK (Frederick et al. Clin Cancer Res. 2013; 19(5):1225-31; Wilmott et al. Clin Cancer Res. 2012;18(5):1386-94).

Without being limited by theory, it is believed that in some embodiments, the use of a BRAF inhibitor (eg, dabrafenib) or a MEK inhibitor (eg, trametinib), alone or in combination, may improve response to an immunotherapeutic agent, for example, by altering the immune microenvironment (Schilling et al Ann Oncol 2014 25(3):747-53 Liu et al Clin Cancer Res 2015 1;21(7):1639-51 Kakavand et al Clin Cancer Res 2015 21(14 ):3140-8).

In some embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 600 mg, e.g., 300 mg to 500 mg (e.g., 400 mg) once every four weeks, or once every eight weeks. An analysis of the population pharmacokinetics (PK) of the anti-PD-1 antibody molecule described in this application showed that changes in exposure due to different body weights of patients are minimal in the expected body weight range of 30-150 kg for a group of patients, which confirms the choice of a scheme for introducing a constant doses. A constant dose of approximately 400 mg once every four weeks or approximately 300 mg once every three weeks is expected to provide a higher mean Ctrough at steady state than the ex vivo EC50 for antigen-stimulated production of IL-2, a translational biomarker for PD-blockade. 1 (Patnaik et al. Clin Cancer Res. 2015; 21(19):4286-93). Based at least in part on the observed safety profile and expected Ctrough values, a dose of approximately 400 mg once every four weeks is expected to be a safe and effective dose. Without being limited by theory, it is believed that in some embodiments, increasing the interval between administrations of the anti-PD-1 antibody molecule (eg, up to eight weeks or more) may make the combination more tolerable. Given the long half-life of the anti-PD-1 antibody molecule described herein, Ctrough values at week 8 are expected to exceed the ex vivo EC50 for antigen-stimulated IL-2 production.

In some embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg) once every four weeks, e.g., intravenously, e.g., for 15 -120 minutes (for example, approximately 30 minutes). In other embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg) once every eight weeks, e.g., intravenously, e.g., for 15 -120 minutes (for example, approximately 30 minutes). In some embodiments, dosing may be interrupted for up to 12 weeks, such as up to 8 weeks or up to 4 weeks.

In some embodiments, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered to a subject in the dosage form described herein. In some embodiments, the dosage form is a lyophilized dosage form. In other embodiments, the dosage form is a reconstituted dosage form, such as reconstituted from a lyophilized dosage form. In other embodiments, the dosage form is a liquid dosage form. In some embodiments, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is reconstituted using sterile water for injection.

In some embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg), e.g., once every four weeks, e.g. intravenously, and: (i) a BRAF inhibitor or dabrafenib is administered at a dose of 50 mg to 300 mg (e.g. 100 mg to 200 mg, e.g. 150 mg), e.g. twice a day, e.g., orally, or in a total daily dose 100 mg to 600 mg (e.g., 200 mg to 400 mg, e.g., in a total daily dose of 300 mg), e.g., orally, (ii) a MEK inhibitor or trametinib is administered at a dose of 0.5 mg to 4 mg (e.g., , 1 mg to 3 mg, eg in a dose of 2 mg), eg, once a day, eg, orally; or (iii) - (i) and (ii).

In some embodiments, the combination comprises a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule, a BRAF inhibitor or dabrafenib, and a MEK inhibitor or trametinib. In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered into 300 mg to 500 mg (eg, 400 mg), eg, once every four weeks, eg, IV; BRAF inhibitor or dabrafenib, 50 mg to 300 mg (eg, 100 mg to 200 mg e.g. at a dose of 150 mg), e.g. twice a day, e.g. orally, or in a total daily dose of 100 mg to 600 mg (e.g. 200 mg to 400 mg, e.g. in a total daily dose of 300 mg) eg, orally; and the MEK inhibitor or trametinib is administered at a dose of 0.5 mg to 4 mg (eg, 1 mg to 3 mg, eg, 2 mg dose), eg, once a day, eg, orally.

In some embodiments, the administration of the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is started on the same day as the administration of the BRAF inhibitor or dabrafenib, the MEK inhibitor or trametinib, or both. In other embodiments, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered following administration of a BRAF inhibitor or dabrafenib, a MEK inhibitor, or trametinib, or both, for about four weeks or more. For example, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) may be started on Day 29 when a BRAF inhibitor or dabrafenib, a MEK inhibitor, or trametinib, or both, is started on Day 1. As an example, a BRAF inhibitor or dabrafenib may be administered twice daily in a schedule for Days 1-28 of a 28 day course, and a MEK inhibitor or trametinib may be administered in a schedule for Days 1-28 of a 28 day course.

In some embodiments, the PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg), for example, once every eight weeks, for example, intravenously, and: (i) a BRAF inhibitor or dabrafenib is administered at a dose of 50 mg to 300 mg (e.g. 100 mg to 200 mg, e.g. 150 mg), e.g. twice a day, e.g., orally, or in a total daily dose 100 mg to 600 mg (e.g., 200 mg to 400 mg, e.g., in a total daily dose of 300 mg), e.g., orally, (ii) a MEK inhibitor or trametinib is administered at a dose of 0.5 mg to 4 mg (e.g., , 1 mg to 3 mg, eg in a dose of 2 mg), eg, once a day, eg, orally; or (iii) - (i) and (ii).

In some embodiments, the combination comprises a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule, a BRAF inhibitor or dabrafenib, and a MEK inhibitor or trametinib. In some embodiments, a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered into 300 mg to 500 mg (eg, 400 mg), eg, once every eight weeks, eg, IV; BRAF inhibitor or dabrafenib given 50 mg to 300 mg (eg, 100 mg to 200 mg e.g. at a dose of 150 mg), e.g. twice a day, e.g. orally, or in a total daily dose of 100 mg to 600 mg (e.g. 200 mg to 400 mg, e.g. in a total daily dose of 300 mg) , eg, orally; and the MEK inhibitor or trametinib is administered at a dose of 0.5 mg to 4 mg (eg, 1 mg to 3 mg, eg, 2 mg dose), eg, once a day, eg, orally.

In some embodiments, the combination is used to treat cancer, eg, melanoma, with elevated levels (eg, serum levels) of LDH, eg, compared to a reference LDH level, eg, LDH levels in a healthy subject. Although, without being limited by theory, it is believed that in some embodiments, serum LDH is an independent prognostic factor in melanoma and correlates with a poor prognosis, as evidenced by the poor survival of patients with advanced melanoma compared to patients with LDH levels in the normal range. Studies have also shown that elevated LDH was a negative predictor of response to therapy and other measures of efficacy (Sirott et al. Cancer. 1993; 72(10):3091-8; Eton et al. J Clin Oncol. 1998; 16(3): 1103-11; Balch et al J Clin Oncol 2001; 19(16):3622-34). The population of patients with elevated LDH levels represents an area of unmet medical need that warrants the use of the treatment option (eg combination therapy) described herein. In some embodiments, the cancer, such as melanoma, also has a BRAF V600 mutation.

In some embodiments, the subject's LDH level is at least 2, 3, 4, or 5 times higher than a reference level, such as the reference level described herein. In some embodiments, baseline LDH is between 1× upper limit of normal (ULN) and 2× ULN. In other embodiments, the baseline LDH level is greater than 2x ULN, eg, greater than 3x ULN, 4x ULN, or 5x ULN. In some embodiments, the LDH level is greater than about 100-350 (eg, about 105-333) IU/L (international units per liter). In some embodiments, the implementation of the level of LDH is determined in serum. LDH assays are described, for example, in Ferri FF, ed. Ferri's Clinical Advisor 2014. Philadelphia: Pa: Elsevier Mosby; 2014: Section IV - Laboratory studies and interpretation of results. In some embodiments, the subject has an ECOG status score selected from 0, 1, or 2. Methods for determining an ECOG status score are described in, for example, Oken et al. Am J Clin Oncol. 1982; 5:649-655.

In some embodiments, the combination is used to treat cancer (eg, melanoma) with a BRAF mutation, eg, the BRAF mutation described herein. In some embodiments, the BRAF mutation is a V600 mutation. In some embodiments, the BRAF mutation is located in the activating segment of the kinase domain. In other embodiments, the BRAF mutation results in increased kinase activity and is optionally transformative in vitro . In some embodiments, the BRAF mutation is selected from a V600E mutation, a V600K mutation, or a V600D mutation. In some embodiments, the BRAF mutation is a V600E mutation. In other embodiments, the combination is used to treat cancer (eg, melanoma) other than wild-type BRAF cancer (eg, wild-type BRAF melanoma). In some embodiments, the BRAF mutation is a mutation that is sensitive to or responsive to a BRAF inhibitor, a MEK inhibitor, or both.

Examples of BRAF mutations include, but are not limited to, BRAF c.1779_1780delTGinsGA (D594N), BRAF c.1780G>C (D594H), BRAF c.1780G>A (D594N), BRAF c.1781A>G (D594G), BRAF c. 1781A>T (D594V), BRAF c.1782T>A (D594E), BRAF c.1782T>G (D594E), BRAF c.1789C>G (L597V), BRAF c.1789_1790delCTinsTC (L597S), BRAF c.1790T> BRAF c.1798_1799delGTinsAA (V600K), BRAF c.1798_1799delGTinsAG (V600R), BRAF c.1799T>A (V600E) , BRAF c.1799T>G (V600G), BRAF c.1799_1800delTGinsAT (V600D), BRAF c.1799_1800delTGinsAA (V600E), or BRAF c.1801A>G (K601E). Other examples of BRAF mutations also include BRAF fusion, for example as described in Botton et al. Pigment Cell Melanoma Res. 2013; 26(6):845-51; Hutchinson et al. Clinic Cancer Res. 2013; 19(24):6696-702).

In some embodiments, the PD-1 inhibitor is an anti-PD-L1 antibody molecule.

Other examples of BRAF inhibitors that may be used in the combination described herein include, but are not limited to, vemurafenib (also known as RG7204 or PLX4032), GDC-0879, PLX-4720, sorafenib, or LGX818.

Other examples of MEK inhibitors that may be used in the combination described herein include, but are not limited to, cobimetinib (also known as XL518), binimetinib (also known as MEK162), selumetinib, 32590PD-1, CI-1040, 03590PD- 1 or TAK-733.

In some embodiments, the combination described herein is also used in combination with surgery, a chemotherapeutic agent, radiation therapy, immunotherapy (e.g., anti-CTLA-4 antibody molecule therapy, anti-PD-L1 antibody molecule, interferon, interleukin (e.g., IL -2) or TNF) or targeted therapy (eg, an oncolytic virus or an angiogenesis inhibitor).

Pharmaceutical compositions and kits

In another aspect, the present invention provides compositions, eg, pharmaceutically acceptable compositions, which comprise the antibody molecule described herein in dosage form with a pharmaceutically acceptable carrier. As used herein, a "pharmaceutically acceptable carrier" includes any possible solvents, dispersion media, isotonic and absorption delaying agents, and the like, which are physiologically compatible. The carrier may be suitable for intravenous, intramuscular, subcutaneous, parenteral, rectal, spinal, or epidermal administration (eg, by injection or infusion).

Compositions according to the present invention may be in various forms. They include, for example, liquid, semi-solid and solid dosage forms such as liquid solutions (eg solutions for injection or infusion), dispersions or suspensions, liposomes and suppositories. The preferred form depends on the intended route of administration and therapeutic application. Typical preferred compositions are in the form of injection or infusion solutions. The preferred route of administration is parenteral (eg, 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.

The phrases "parenteral administration" and "parenterally administered" as used herein means modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion.

Therapeutic compositions generally must be sterile and stable under the conditions of manufacture and storage. The composition may 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 active compound (i.e., antibody or antibody portion) in the required amount in a suitable solvent with one or a combination of the ingredients listed above, followed by filtered sterilization, if necessary. Typically, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains the basic dispersion medium and the other necessary components listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of manufacture are vacuum drying and freeze-drying, which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterilized filtered solution. The desired fluidity of the solution can be maintained, for example, when using a coating such as lecithin, by maintaining the required particle size in the case of a dispersion and when using surfactants. Prolonged absorption of injectable compositions can be achieved by including in the composition a substance that delays absorption, such as monostearate salts and gelatin.

The PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) or composition described herein may be formulated into a dosage form (eg, dosage formulation or dosage form) suitable for administration (eg, intravenous administration) to a subject, as described in this application.

In some embodiments, the dosage form is a drug substance composition. In other embodiments, the dosage form is a lyophilized dosage form, eg, lyophilized or dried from a drug substance composition. In other embodiments, the dosage form is a reconstituted dosage form, such as reconstituted from a lyophilized dosage form. In other embodiments, the dosage form is a liquid dosage form.

In some embodiments, the dosage form is a drug substance composition. In some embodiments, the dosage form (eg, drug substance composition) includes a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) and a buffering agent.

In some embodiments, the dosage form (e.g., drug substance composition) comprises a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) present at a concentration of 10-50 mg/mL, e.g., 15-50 mg/mL, 20- 45 mg/ml, 25-40 mg/ml, 30-35 mg/ml, 25-35 mg/ml, or 30-40 mg/ml, e.g. 15 mg/ml, 20 mg/ml, 25 mg/ml , 30 mg/ml, 33.3 mg/ml, 35 mg/ml, 40 mg/ml, 45 mg/ml or 50 mg/ml. In some embodiments, the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is present at a concentration of 30-35 mg/mL, such as 33.3 mg/mL.

In some embodiments, the dosage form (eg, drug substance composition) includes a buffering agent comprising histidine (eg, a histidine buffer). In some embodiments, the buffering agent (e.g., histidine buffer) is present at a concentration of 1 mM to 20 mM, e.g., 2 mM to 15 mM, 3 mM to 10 mM, 4 mM to 9 mM, 5 mM to 8 mM mM or from 6 mM to 7 mM, e.g. mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM or 20 mM. In some embodiments, the buffering agent (eg, histidine buffer) is present at a concentration of 6 mM to 7 mM, such as 6.7 mM. In other embodiments, the buffering agent (e.g., histidine buffer) has a pH of 4-7, e.g., 5-6, e.g., 5, 5.5, or 6. In some embodiments, the buffering agent (e.g., histidine buffer) has a pH of 5-6 , for example 5.5. In some embodiments, the buffering agent comprises histidine at a concentration of 6 mM to 7 mM (eg, 6.7 mM) and has a pH of 5-6 (eg, 5.5).

In some embodiments, the dosage form (eg, drug substance composition) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 30-35 mg/mL, eg, 33.3 mg/mL; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5).

In some embodiments, the dosage form (eg, drug substance composition) further includes a carbohydrate. In some embodiments, the carbohydrate is sucrose. In some embodiments, the carbohydrate (e.g., sucrose) is present at a concentration of 50 mM to 150 mM, e.g., 25 mM to 150 mM, 50 mM to 100 mM, 60 mM to 90 mM, 70 mM to 80 mM, or 70 mM to 75 mM, e.g. 25 mM, 50 mM, 60 mM, 70 mM, 73.3 mM, 80 mM, 90 mM, 100 mM or 150 mM. In some embodiments, the implementation of the dosage form includes a carbohydrate or sucrose present in a concentration of from 70 mm to 75 mm, for example, 73.3 mm.

In some embodiments, the dosage form (eg, drug substance composition) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 30-35 mg/ml, eg, 33.3 mg/ml; a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); and carbohydrate or sucrose present at a concentration of 70 mM to 75 mM, eg 73.3 mM.

In some embodiments, the dosage form (eg, drug substance composition) further includes a surfactant. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the surfactant or polysorbate 20 is present at a concentration of 0.005-0.025% (w/w), such as 0.0075-0.02% or 0.01 -0.015% (w/w), such as 0.005%, 0.0075%, 0.01%, 0.013%, 0.015%, or 0.02% (w/w). In some embodiments, the dosage form includes a surfactant or polysorbate 20 present at a concentration of 0.01-0.015%, such as 0.013% (w/w).

In some embodiments, the dosage form (eg, drug substance composition) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 30-35 mg/ml, eg, 33.3 mg/ml; a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); and a surfactant or polysorbate 20 present at a concentration of 0.01-0.015%, for example 0.013% (w/w).

In some embodiments, the dosage form (eg, drug substance composition) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 30-35 mg/ml, eg, 33.3 mg/ml; a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); a carbohydrate or sucrose present in a concentration of 70 mM to 75 mM, eg 73.3 mM; and a surfactant or polysorbate 20 present at a concentration of 0.01-0.015%, for example 0.013% (w/w).

In some embodiments, the dosage form (eg, drug substance composition) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 33.3 mg/mL; a buffer that includes histidine at a concentration of 6.7 mM and has a pH of 5.5; sucrose present at a concentration of 73.3 mM; and polysorbate 20 present at a concentration of 0.013% (w/w).

In some embodiments, the dosage form is a lyophilized dosage form. In some embodiments, the lyophilized dosage form is lyophilized from the drug substance composition described in this application. For example, 2-5 ml, eg 3-4 ml, eg 3.6 ml, of the drug substance composition described in this application may be filled into a container (eg, vial) and lyophilized.

In some embodiments, the dosage form is a reconstituted dosage form. For example, a reconstituted dosage form can be prepared by dissolving the lyophilized dosage form in a diluent, with the protein dispersed in the reconstituted dosage form. In some embodiments, the lyophilized dosage form is reconstituted in 0.5-2 ml, eg 1 ml, of water or injection buffer. In some embodiments, the lyophilized dosage form is reconstituted with 1 ml of water for injection, for example, at a clinical center.

In some embodiments, the dosage form (eg, reconstituted dosage form) includes a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) and a buffering agent.

In some embodiments, the dosage form (e.g., reconstituted dosage form) comprises a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) present at a concentration of 20 mg/mL to 200 mg/mL, such as 50 mg/mL up to 150 mg/ml, 80 mg/ml to 120 mg/ml or 90 mg/ml to 110 mg/ml, e.g. 50 mg/ml, 60 mg/ml, 70 mg/ml, 80 mg/ml, 90 mg/ml, 100 mg/ml, 110 mg/ml, 120 mg/ml, 130 mg/ml, 140 mg/ml, 150 mg/ml, 160 mg/ml, 170 mg/ml, 180 mg/ml, 190 mg/ml or 200 mg/ml. In some embodiments, the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is present at a concentration of 80-120 mg/mL, such as 100 mg/mL.

In some embodiments, the dosage form (eg, reconstituted dosage form) includes a buffering agent comprising histidine (eg, a histidine buffer). In some embodiments, the buffering agent (e.g., histidine buffer) is present at a concentration of 5 mM to 100 mM, e.g., 10 mM to 50 mM, 15 mM to 25 mM, e.g., 5 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM or 100 mM. In some embodiments, the implementation of the buffer substance (eg, his-tag buffer) is present at a concentration of from 15 mm to 25 mm, for example, 20 mm. In other embodiments, the buffering agent (e.g., histidine buffer) has a pH of 4-7, e.g., 5-6, e.g., 5, 5.5, or 6. In some embodiments, the buffering agent (e.g., histidine buffer) has a pH of 5-6, for example 5.5. In some embodiments, the buffering agent comprises histidine at a concentration of 15 mM to 25 mM (eg, 20 mM) and has a pH of 5-6 (eg, 5.5).

In some embodiments, the dosage form (eg, reconstituted dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 80-120 mg/mL, eg, 100 mg/mL; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5).

In some embodiments, the dosage form (eg, reconstituted dosage form) further comprises a carbohydrate. In some embodiments, the carbohydrate is sucrose. In some embodiments, the carbohydrate (e.g., sucrose) is present at a concentration of 100 mM to 500 mM, e.g., 150 mM to 400 mM, 175 mM to 300 mM, or 200 mM to 250 mM, e.g., 150 mM, 160 mM , 170 mM, 180 mM, 190 mM, 200 mM, 210 mM, 220 mM, 230 mM, 240 mM, 250 mM, 260 mM, 270 mM, 280 mM, 290 mM or 300 mM In some embodiments, the dosage form comprises a carbohydrate or sucrose present at a concentration of 200 mM to 250 mM, such as 220 mM.

In some embodiments, the dosage form (eg, reconstituted dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 80-120 mg/mL, eg, 100 mg/mL; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); and carbohydrate or sucrose, which are present at a concentration of 200 mM to 250 mM, eg 220 mM.

In some embodiments, the dosage form (eg, reconstituted dosage form) further comprises a surfactant. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the surfactant or polysorbate 20 is present at a concentration of 0.01-0.1% (w/w), such as 0.02-0.08% , 0.025-0.06% or 0.03-0.05% (w/w), e.g. 0.01%, 0.025%, 0.03%, 0.04%, 0.05%, 0.06 %, 0.07%, 0.08%, 0.09% or 0.1% (w/w). In some embodiments, the dosage form includes a surfactant or polysorbate 20 present at a concentration of 0.03-0.05%, such as 0.04% (w/w).

In some embodiments, the dosage form (eg, reconstituted dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 80-120 mg/mL, eg, 100 mg/mL; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); and a surfactant or polysorbate 20 present at a concentration of 0.03-0.05%, for example 0.04% (w/w).

In some embodiments, the dosage form (eg, reconstituted dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 80-120 mg/mL, eg, 100 mg/mL; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); carbohydrate or sucrose present in a concentration of 200 mM to 250 mM, for example 220 mM; and a surfactant or polysorbate 20 present at a concentration of 0.03-0.05%, for example 0.04% (w/w).

In some embodiments, the dosage form (eg, reconstituted dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 100 mg/mL; and a buffer that includes histidine at a concentration of 6.7 mM and has a pH of 5.5; sucrose present at a concentration of 220 mM; and polysorbate 20 present at a concentration of 0.04% (w/w).

In some embodiments, the dosage form is reconstituted such that a withdrawal volume of at least 1 ml (e.g., at least 1.5 ml, 2 ml, 2.5 ml, or 3 ml) of the reconstituted dosage form can be withdrawn from the container (e.g., , vial) containing the reconstituted dosage form. In some embodiments, the dosage form is reconstituted and/or withdrawn from a container (eg, vial) at a clinical center. In some embodiments, the dosage form (eg, reconstituted dosage form) is administered in an infusion bag, eg, within 1 hour (eg, within 45 minutes, 30 minutes, or 15 minutes) prior to infusion to the patient.

In some embodiments, the dosage form is a liquid dosage form. In some embodiments, a liquid dosage form is made by diluting the composition of the drug substance described in this application. For example, the drug substance composition can be diluted, for example, 10-30 mg/ml (eg, 25 mg/ml) solution containing one or more excipients (eg, concentrated excipients). In some embodiments, the solution includes one, two, or all of histidine, sucrose, or polysorbate 20. In some embodiments, the solution includes the same excipient(s) as the drug substance composition. Examples of excipients include, but are not limited to, an amino acid (eg, histidine), a carbohydrate (eg, sucrose), or a surfactant (eg, polysorbate 20). In some embodiments, the liquid dosage form is not a reconstituted lyophilized dosage form. In other embodiments, the liquid dosage form is a reconstituted lyophilized dosage form. In some embodiments, the dosage form is stored as a liquid. In other embodiments, the dosage form is formulated into a liquid and then dried, such as by lyophilization or spray drying, prior to storage.

In some embodiments, the dosage form (e.g., liquid dosage form) comprises a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) present at a concentration of 5 mg/mL to 50 mg/mL, such as 10 mg/mL up to 40 mg/ml, 15 mg/ml to 35 mg/ml or 20 mg/ml to 30 mg/ml, e.g. 5 mg/ml, 10 mg/ml, 15 mg/ml, 20 mg/ml, 25 mg/ml, 30 mg/ml, 35 mg/ml, 40 mg/ml, 45 mg/ml or 50 mg/ml. In some embodiments, the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is present at a concentration of 20-30 mg/mL, such as 25 mg/mL.

In some embodiments, the implementation of the dosage form (eg, liquid dosage form) includes a buffer substance, including histidine (eg, histidine buffer). In some embodiments, the buffering agent (e.g., histidine buffer) is present at a concentration of 5 mM to 100 mM, e.g., 10 mM to 50 mM, 15 mM to 25 mM, e.g., 5 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM or 100 mM. In some embodiments, the buffering agent (eg, histidine buffer) is present at a concentration of 15 mM to 25 mM, such as 20 mM. In other embodiments, the buffering agent (e.g., histidine buffer) has a pH of 4-7, e.g., 5-6, e.g., 5, 5.5, or 6. In some embodiments, the buffering agent (e.g., histidine buffer) has a pH of 5-6, for example 5.5. In some embodiments, the buffering agent comprises histidine at a concentration of 15 mM to 25 mM (eg, 20 mM) and has a pH of 5-6 (eg, 5.5).

In some embodiments, the dosage form (eg, liquid dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 20-30 mg/ml, eg 25 mg/ml; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5).

In some embodiments, the dosage form (eg, liquid dosage form) further comprises a carbohydrate. In some embodiments, the carbohydrate is sucrose. In some embodiments, the carbohydrate (e.g., sucrose) is present at a concentration of 100 mM to 500 mM, e.g., 150 mM to 400 mM, 175 mM to 300 mM, or 200 mM to 250 mM, e.g., 150 mM, 160 mM , 170 mM, 180 mM, 190 mM, 200 mM, 210 mM, 220 mM, 230 mM, 240 mM, 250 mM, 260 mM, 270 mM, 280 mM, 290 mM or 300 mM In some embodiments, the dosage form comprises a carbohydrate or sucrose present at a concentration of 200 mM to 250 mM, such as 220 mM.

In some embodiments, the dosage form (eg, liquid dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 20-30 mg/ml, eg 25 mg/ml; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); and carbohydrate or sucrose present at a concentration of 200 mM to 250 mM, eg 220 mM.

In some embodiments, the dosage form (eg, liquid dosage form) further comprises a surfactant. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the surfactant or polysorbate 20 is present at a concentration of 0.01-0.1% (w/w), for example, 0.02-0.08 %, 0.025-0.06% or 0.03-0.05% (w/w), e.g. 0.01%, 0.025%, 0.03%, 0.04%, 0.05%, 0, 06%, 0.07%, 0.08%, 0.09% or 0.1% (w/w). In some embodiments, the dosage form includes a surfactant or polysorbate 20 present at a concentration of 0.03-0.05%, such as 0.04% (w/w).

In some embodiments, the dosage form (eg, liquid dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 20-30 mg/ml, eg 25 mg/ml; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); and a surfactant or polysorbate 20 present at a concentration of 0.03-0.05%, eg 0.04% (w/w).

In some embodiments, the dosage form (eg, liquid dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 20-30 mg/ml, eg 25 mg/ml; and a buffer that includes histidine at a concentration of 6 mM to 7 mM (eg 6.7 mM) and has a pH of 5-6 (eg 5.5); carbohydrate or sucrose present in a concentration of 200 mM to 250 mM, for example 220 mM; and a surfactant or polysorbate 20 present at a concentration of 0.03-0.05%, eg 0.04% (w/w).

In some embodiments, the dosage form (eg, liquid dosage form) comprises a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) present at a concentration of 25 mg/mL; and a buffer that includes histidine at a concentration of 6.7 mM and has a pH of 5.5; sucrose present at a concentration of 220 mM; and polysorbate 20 present at a concentration of 0.04% (w/w).

In some embodiments, 1 ml to 10 ml (e.g., 2 ml to 8 ml, 3 ml to 7 ml, or 4 ml to 5 ml, e.g., 3 ml, 4 ml, 4.3 ml, 4.5 ml, 5 ml or 6 ml) of the liquid dosage form is filled into a container (eg vial). In other embodiments, the liquid dosage form is filled into a container (e.g., vial) such that a withdrawal volume of at least 2 ml (e.g., at least 3 ml, at least 4 ml, or at least 5 ml) of the liquid dosage form can be withdraw from each container (e.g. vial). In some embodiments, the liquid dosage form is diluted from the drug substance composition and/or withdrawn from a container (eg, vial) at the clinical center. In some embodiments, the dosage form (eg, liquid dosage form) is administered in an infusion bag, eg, within 1 hour (eg, within 45 minutes, 30 minutes, or 15 minutes) prior to infusion to the patient.

The dosage form described in this application may be stored in a container. The container used for any of the dosage forms described herein may include, for example, a vial and optionally a stopper, a cap, or both. In some embodiments, the vial is a glass vial, such as a 6R clear glass vial. In other embodiments, the stopper is a rubber stopper, such as a gray rubber stopper. In other embodiments, the cap is a flip cap, such as an aluminum flip cap. In some embodiments, the container includes a 6R clear glass vial, a gray rubber stopper, and an aluminum flip cap. In some embodiments, the container (eg, vial) is a single use container. In some embodiments, 50 mg to 150 mg is present in a container (e.g., vial), e.g., 80 mg to 120 mg, 90 mg to 110 mg, 100 mg to 120 mg, 100 mg to 110 mg, 110 mg to 120 mg, or 110 mg to 130 mg, of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule).

Other examples of buffer substances that can be used in the dosage form described in this application include, without limitation, arginine buffer, citrate buffer or phosphate buffer. Other examples of carbohydrates that can be used in the dosage form described in this application include, without limitation, trehalose, mannitol, sorbitol, or a combination thereof. The dosage form described herein may also contain a tonicity agent, eg sodium chloride, and/or a stabilizing agent, eg an amino acid (eg, glycine, arginine, methionine, or a combination thereof).

The antibody molecules can be administered by various methods known in the art, although for many therapeutic applications the preferred route/method of administration is intravenous injection or infusion. For example, antibody molecules can be administered by intravenous infusion at rates greater than 20 mg/min, such as 20-40 mg/min, and typically greater than or equal to 40 mg/min, to give a dose of approximately 35-440 mg/m ² typically about 70-310 mg/m ² , and more typically about 110-130 mg/m ² . In embodiments, the antibody molecules may be administered by intravenous infusion at a rate of less than 10 mg/min; preferably less than or equal to 5 mg/min to give a dose of about 1-100 mg/m 2 , preferably about 5-50 mg/m ² , about 7-25 mg/m ² , and more preferably about 10 mg/m ² . It will be apparent to a person skilled in the art that the route and/or mode of administration will vary depending on the desired results. In some embodiments, the active compound may be formulated with a carrier that will prevent rapid release of the compound, eg, in a controlled release dosage form, 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 making such compositions are patented or generally known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, JR Robinson, ed., Marcel Dekker, Inc., New York, 1978.

In some embodiments, the antibody molecule may be administered orally, for example, with an inert diluent or a digestible edible carrier. The compound (and other components, if desired) may also be enclosed in a hard or soft shell gelatin capsule, compressed into tablets, or incorporated directly into the subject's diet. For oral therapeutic administration, the compounds may be combined with excipients and administered in the form of oral tablets, buccal tablets, lozenges, capsules, tinctures, suspensions, syrups, cachets, and the like. In order to administer a compound of the invention by routes other than parenteral administration, it may be necessary to coat the compound or administer the compound with a material to prevent its inactivation. Therapeutic compositions can also be administered using medical devices known in the art.

Dosage regimens are selected to provide the optimum desired response (eg, therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as required by the clinical situation. Particularly preferred is the formulation of parenteral compositions in the form of a unit dosage form due to ease of administration and uniformity of dose. Unit dosage form, as used herein, refers to physically discrete units suitable as unit doses for the subjects to be treated, each unit containing a predetermined amount of the active compound calculated to produce the desired therapeutic effect, in combination with the required pharmaceutical carrier. . The specification for unit dosage forms according to the invention is determined by and depends directly on: (a) the unique properties of the active compound and the intended therapeutic effect, and (b) the limitations inherent in the preparation of mixtures of such an active compound for the treatment of sensitivity in patients.

An exemplary non-limiting range for a therapeutically or prophylactically effective amount of an antibody molecule is 0.1-30 mg/kg, more preferably 1-25 mg/kg. Doses and therapeutic regimens for the use of an anti-PD-1 antibody molecule can be determined by those skilled in the art. In some embodiments, the anti-PD-1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously), at a dose of about 1-40 mg/kg, e.g. 1-30 mg/kg, e.g., about 5-25 mg/kg, about 10-20 mg/kg, approximately 1-5 mg/kg, 1-10 mg/kg, 5-15 mg/kg, 10-20 mg/kg, 15-25 mg/kg or approximately 3 mg/kg. The administration schedule may vary, for example, from once a week to once every 2, 3 or 4 weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 10-20 mg/kg biweekly.

As another example, a non-limiting range for a therapeutically or prophylactically effective amount of an antibody molecule is 200-500 mg, more preferably 300-400 mg/kg. Doses and therapeutic regimens of the anti-PD-1 antibody molecule can be determined by those skilled in the art. In some embodiments, the anti-PD-1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously), at a dose (e.g., constant dose) of about 200 mg to 500 mg, for example, about 250 mg to 450 mg, about 300 mg to 400 mg, about 250 mg to 350 mg, about 350 mg to 450 mg, or about 300 mg, or about 400 mg. The administration schedule (eg, fixed dose schedule) may vary, for example, from once a week to once every 2, 3, 4, 5 or 6 weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg to 400 mg once every three or once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every three weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every three weeks. Without being limited by theory, in some embodiments, the implementation of the implementation of a constant or fixed dose may be beneficial to patients, for example, to simplify the supply of drugs and to reduce pharmaceutical errors.

In some embodiments, the clearance (CL) of the anti-PD-1 antibody molecule is from about 6-16 ml/h, e.g., about 7-15 ml/h, about 8-14 ml/h, about 9-12 ml/h, or about 10-11 ml/h, for example about 8.9 ml/h, 10.9 ml/h or 13.2 ml/h.

In some embodiments, the coefficient of influence of weight on CL of an anti-PD-1 antibody molecule is from about 0.4 to 0.7, about 0.5 to 0.6, or 0.7, or less, such as 0.6 or less, or approximately 0.54.

In some embodiments, the volume of distribution at steady state (Vss) of an anti-PD-1 antibody molecule is from about 5-10 V, e.g., about 6-9 V, about 7-8 V, or about 6.5-7.5 V, for example, approximately 7.2 V.

In some embodiments, the half-life of an anti-PD-1 antibody molecule is about 10-30 days, such as about 15-25 days, about 17-22 days, about 19-24 days, or about 18-22 days, such as about 20 days. .

In some embodiments, the Cmin (e.g., for an 80 kg patient) of an anti-PD-1 antibody molecule is at least about 0.4 μg/ml, such as at least about 3.6 μg/ml, such as from about 20 -50 µg/ml, eg about 22-42 µg/ml, about 26-47 µg/ml, about 22-26 µg/ml, about 42-47 µg/ml, about 25-35 µg/ml, about 32 -38 µg/ml, eg about 31 µg/ml or about 35 µg/ml. In one embodiment, Cmin is determined in a patient receiving an anti-PD-1 antibody molecule at a dose of approximately 400 mg once every four weeks. In another embodiment, Cmin is determined in a patient receiving an anti-PD-1 antibody molecule at a dose of approximately 300 mg once every three weeks. In some embodiments, Cmin is at least about 50 times higher, such as at least about 60 times, 65 times, 70 times, 75 times, 80 times, 85 times, 90 times, 95 times, or 100 times, for example, at least about 77 times higher than the EC50 of an anti-PD-1 antibody molecule, for example, as determined based on IL-2 change in an ex vivo SEB assay. In other embodiments, the Cmin is at least 5 times higher, such as at least 6 times, 7 times, 8 times, 9 times, or 10 times, such as at least about 8.6 times, higher than the EC90 of the molecule. anti-PD-1 antibodies, for example, as determined based on the change in IL-2 in an ex vivo SEB assay.

The antibody molecule can be administered by intravenous infusion at a rate greater than 20 mg/min, such as 20-40 mg/min, and typically greater than or equal to 40 mg/min, to give a dose of approximately 35-440 mg/m ² as typically about 70-310 mg/m ² and more typically about 110-130 mg/m ² . In embodiments, a drug administration rate of about 110-130 mg/m ² provides a level of about 3 mg/kg. In other embodiments, the antibody molecule may be administered by intravenous infusion at a rate of less than 10 mg/min, such as less than or equal to 5 mg/min, to give a dose of about 1-100 mg/m ² , such as about 5-50 mg/m ² , about 7-25 mg/m ² or about 10 mg/m ² . In some embodiments, the antibody is administered by infusion over about 30 minutes. It should be noted that dosage values may vary depending on the type and severity of the condition to be alleviated. It should also be understood that for any given subject, specific administration schedules will need to be adjusted over time according to individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that the dosage ranges given herein are only exemplary and should not be construed as limiting the scope or practical application of the claimed composition.

Pharmaceutical compositions of the invention may include a "therapeutically effective amount" or "prophylactically effective amount" of an antibody or antibody portion of the invention. "Therapeutically effective amount" refers to an amount that, in the required doses and for the required periods of time, effectively provides the desired therapeutic result. The therapeutically effective amount of the modified antibody or antibody fragment may vary depending on factors such as the pathological condition, age, sex, and weight of the patient, as well as the ability of the antibody or antibody portion to elicit the desired response in the patient. A therapeutically effective amount is also one in which the therapeutically beneficial effects of the modified antibody or antibody fragment outweigh any toxic or deleterious effects thereof. A "therapeutically effective dose" preferably inhibits a measurable parameter, e.g., tumor growth rate, by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and even more preferably at least at least approximately 80% compared with untreated subjects. The ability of a compound to inhibit a measurable parameter, such as cancer, can be assessed using an animal model system that predicts efficacy against human tumors. Alternatively, this property of the composition can be assessed by testing the ability of the compound to inhibit, such inhibition being tested using in vitro alyses known to the person skilled in the art.

"Prophylactically effective amount" refers to the amount, in the required doses and for the required periods of time, which effectively provides the desired prophylactic result. Generally, since a prophylactic dose is administered to subjects prior to or at an early stage of the disease, the prophylactically effective amount will be less than the therapeutically effective amount.

Also within the scope of the present invention is a kit comprising an antibody molecule as described herein. The kit may include one or more other elements, including: instructions for use, other reagents, for example, a label, a therapeutic agent or a substance used to complex or otherwise cross-link an antibody with a label or therapeutic agent, or a radioprotective composition; devices or other materials for obtaining antibodies for administration; pharmaceutically acceptable carriers; and devices or other materials for administration to a subject.

Combination Therapy Applications

Combinations, such as anti-PD-1 antibody molecules, disclosed herein have diagnostic as well as therapeutic and prophylactic uses in vitro and in vivo . For example, these molecules can be administered to cells in culture, in vitro or ex vivo , or to a subject, such as a human, for the treatment, prevention and/or diagnosis of various disorders such as various forms of cancer and infectious diseases.

Thus, in one aspect of the invention, there is provided a method for modifying an immune response in a subject, comprising administering to the subject a combination described herein, resulting in a modification of the immune response in the subject. In one embodiment, there is an enhancement, stimulation or upregulation of the immune response.

As used in this application, the term "subject" includes human and non-human animals. In one embodiment, the subject is a human, eg, a patient, having a disorder or condition characterized by abnormal PD-1 function. The term "non-human animals" 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 patient in need of an enhanced immune response. In one embodiment, the subject has an immunodeficiency state, for example, the subject is undergoing chemotherapy or radiation therapy, or has previously undergone such treatment. Alternatively, or in combination, the subject is or is at risk of developing immunodeficiency as a result of the infection. The methods and compositions described herein are suitable for the treatment of patients suffering from a disease that can be treated by enhancing a T-cell mediated immune response. For example, the methods and compositions described herein can enhance various immune activities. In one embodiment, the subject has an increased number and/or activity of tumor infiltrating T lymphocytes (TILs). In another embodiment, the subject has increased expression or activity of interferon gamma (IFN-γ). In yet another embodiment, the subject has reduced expression or activity of PD-L1.

Therapeutic Applications

Blockade of PD-1 can enhance the immune response against cancer cells in a subject. The PD-1 ligand, PD-L1, is not expressed in normal human cells, but is widely present in various human cancers (Dong et al. (2002) Nat Med 8:787-9). Interaction between PD-1 and PD-L1 can cause a decrease in tumor-infiltrating lymphocytes, a decrease in T-cell receptor-mediated proliferation, and/or immune escape of cancer cells (Dong et al. (2003) J Mol Med 81:281-7; Blank et al (2005) Cancer Immunol Immunother 54:307-314 Konishi et al (2004) Clin Cancer Res 10:5094-100). Immune suppression can be reversed by inhibiting the local interaction of PD-1 with PD-L1; this effect is additive if the interaction of PD-1 with PD-L2 is also blocked (Iwai et al. (2002) PNAS 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66 ). Thus, inhibition of PD-1 can lead to an increase in the immune response.

In one aspect, the present invention relates to in vivo treatment of a subject with an anti-PD-1 antibody molecule that inhibits or reduces the growth of cancerous tumors. Anti-PD-1 antibody can be used alone to inhibit the growth of cancerous tumors. Alternatively, an anti-PD-1 antibody may be used in combination with one or more of: standard of care therapy (e.g., for cancer or infectious diseases), another antibody or antigen-binding fragment thereof, an immunomodulator (e.g., an activator of a costimulatory molecule or an inhibitor of an inhibitory molecule) , vaccines, such as a therapeutic cancer vaccine, or other forms of cellular immunotherapy, as described below.

Thus, in one embodiment, the invention provides a method for inhibiting tumor cell growth in a subject, comprising administering to the subject a therapeutically effective amount of an anti-PD-1 antibody molecule described herein.

In one embodiment, the methods are suitable for in vivo treatment of cancer. To achieve an antigen-specific improvement in immunity, the anti-PD-1 antibody molecule can be administered together with the target antigen. When administering anti-PD-1 antibodies in combination with one or more agents, the combination may be administered in any order or simultaneously.

In another aspect, a method is provided for treating a subject, eg, reducing or alleviating a hyperproliferative condition or disorder (eg, cancer), eg, a solid tumor, hemoblastosis, soft tissue tumor, or metastatic lesion in a subject. The method includes administering to the subject one or more of the combinations disclosed in this application.

As used in this application, the term "cancer" includes all types of malignant neoplasms or oncogenic processes, metastatic tissues or cells, tissues or organs with malignant transformation, regardless of histological type or stage of invasiveness. Examples of cancers include, but are not limited to, solid tumors, hemoblastoses, soft tissue tumors, and metastatic lesions. Examples of solid tumors include malignancies such as sarcomas and carcinomas (including adenocarcinomas and squamous cell carcinomas), various organ systems such as those affecting the liver, lung, breast, lymphoid tissue, gastrointestinal tract (such as the colon), genitourinary tract (eg, kidneys, urothelial cells), prostate, and pharynx. Adenocarcinomas include malignancies such as most colon cancers, rectal cancer, renal cell carcinoma, liver cancer, non-small cell lung carcinoma, small intestine cancer, and cancer of the esophagus. Squamous cell carcinomas include malignancies in, for example, the lung, esophagus, skin, head and neck, mouth, anus, and cervix. In one embodiment, the cancer is melanoma, eg, advanced stage melanoma. Metastatic lesions in the above forms of cancer can also be treated or prevented using the methods and compositions of the invention.

Examples of cancers, the growth of which can be inhibited using the antibody molecules disclosed in this application include cancers that are usually sensitive to immunotherapy. Non-limiting examples of preferred cancers for treatment include melanoma (eg, metastatic malignant melanoma), kidney cancer (eg, clear cell carcinoma), prostate cancer (eg, hormone refractory adenocarcinoma of the prostate), breast cancer, colon cancer, and lung cancer (eg. , non-small cell lung cancer). Additionally, refractory or recurrent cancers can be treated with the antibody molecules described herein.

Examples of other forms of cancer 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 cancer, cancer stomach, liposarcoma, testicular cancer, uterine cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Merkel cell carcinoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, cancer thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemias, including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, childhood solid tumors, lymphocytic lymphoma, bladder cancer, multiple myeloma, myelodysplastic syndrome, kidney or ureter cancer, renal pelvis carcinoma neoplasms of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, tumor of the spinal canal, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell carcinoma, T-cell lymphoma, forms of cancer caused by environmental factors , including asbestos-related cancers (eg, mesothelioma) and combinations of these cancers. In some embodiments, the cancer is skin cancer, such as Merkel cell cancer or melanoma. In one embodiment, the cancer is Merkel cell cancer. In other embodiments, the cancer is melanoma. In other embodiments, the cancer is breast cancer, such as triple-negative breast cancer (TNBC) or HER2-negative breast cancer. In other embodiments, the cancer is kidney cancer, eg, renal cell carcinoma (eg, clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC)). In other embodiments, the cancer is thyroid cancer, such as anaplastic thyroid carcinoma (ATC). In other embodiments, the cancer is a neuroendocrine tumor (NET), such as an atypical pulmonary carcinoid tumor or a NET in the pancreas, gastrointestinal (GI) tract, or lung. In some embodiments, the cancer is lung cancer, eg, non-small cell lung cancer (NSCLC) (eg, squamous NSCLC or non-squamous NSCLC). In some embodiments, the cancer is a leukemia (eg, acute myeloid leukemia (AML), eg, relapsed or refractory AML, or de novo AML). In some embodiments, the cancer is a myelodysplastic syndrome (MDS) (eg, high risk MDS).

In some embodiments, the cancer is selected from lung cancer, squamous cell lung cancer, melanoma, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, ER+ breast cancer, IM-TN breast cancer, colon and rectal cancer , colon and rectal cancer with high microsatellite instability, EBV+ gastric cancer, pancreatic cancer, thyroid cancer, hemoblastosis, non-Hodgkin's lymphoma or leukemia, or cancer metastatic lesion. In some embodiments, the cancer is selected from non-small cell lung cancer (NSCLC), NSCLC adenocarcinoma, NSCLC squamous cell carcinoma, hepatocellular carcinoma.

Treatment of metastatic cancers, for example, metastatic cancers that express PD-L1 (Iwai et al. (2005) Int. Immunol. 17:133-144), can be carried out using the antibody molecules described in this application. In one embodiment, the cancer expresses elevated levels of PD-L1, IFNγ, and/or CD8.

Without being limited by theory, in some embodiments, a patient is more likely to respond to treatment with an immunomodulator (optionally in combination with one or more agents as described herein) if the patient has a highly expressed PD-L1 cancer, and/or if the cancerous tumor is infiltrated with antitumor immune cells such as TIL cells. Antitumor immune cells may be positive for CD8, PD-L1 and/or IFN-γ; thus, CD8, PD-L1 and/or IFN-γ levels can serve as indicators of TIL levels in the microenvironment. In some embodiments, the cancer microenvironment is indicated as a triple positive phenotype for PD-L1/CD8/IFN-γ.

Thus, in some aspects, the present application provides methods for determining if 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, for example, two or all three of these markers, then the patient is administered a therapeutically effective amount of the anti-PD-1 antibody molecule, optionally in combination with one or more other immunomodulators or antineoplastic agents.

In the following indications, most 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 cancer (eg, MSIhi and/or EBV+); CRC (eg, MSIhi), nasopharyngeal cancer (NPC), cervical cancer (eg, squamous cell), thyroid cancer, eg, papillary thyroid cancer, eg, anaplastic thyroid carcinoma; skin cancer (for example, Merkel cell cancer or melanoma); breast cancer (eg, TN breast cancer); and DLBCL (diffuse large B-cell lymphoma). Typically, in breast cancer and usually in colon cancer, a moderate proportion of patients are triple positive for PD-L1/CD8/IFN-γ. In the following indications, a small proportion of patients are triple positive for PD-L1/CD8/IFN-γ: ER+ breast cancer and pancreatic cancer. These results are discussed below in Example 4. Regardless of whether a large or small proportion of patients are triple positive for these markers, screening patients for such markers identifies a subset of patients who are particularly likely to respond effectively to PD-1 antibody therapy (e.g., blocking PD-1 antibody), optionally in combination with one or more other immunomodulators (e.g., an anti-TIM-3 antibody molecule, an anti-LAG-3 antibody molecule, or an anti-PD-L1 antibody molecule) and/or antineoplastic agents, for example, those listed in Table 7 and disclosed in the publications listed in Table 7.

In some embodiments, the cancer sample is classified as triple positive for PD-Ll/CD8/IFN-γ. This definition can be roughly divided into two threshold conditions: when an individual cell is classified as positive, and when the sample as a whole is classified as positive. First, you can measure, in a single cell, the level of PD-L1, CD8 and/or IFN-γ. In some embodiments, a cell that is positive for one or more of these markers is a cell that has a higher level of the marker than a control cell or reference value. For example, in some embodiments, a high level of PD-L1 in a given cell is higher than a level of PD-L1 in a corresponding non-cancerous tissue in a patient. As another example, in some embodiments, a high level of CD8 or IFN-γ in a given cell is the level of that protein normally seen in TIL. Secondly, it is also possible to measure the percentage of cells in the sample that are positive for PD-L1, CD8 and/or IFN-γ. (One cell does not have to express all three markers). In some embodiments, a triple positive sample is a sample that contains a high percentage of cells, such as higher than a reference value or higher than a control sample, that are positive for the indicated markers.

In other embodiments, total 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 may be the level of such a protein typically seen in a TIL-infiltrated tumor. Similarly, a high level of PD-L1 may be the level of such a protein typically seen in a tumor sample, eg, the tumor microenvironment.

The identification of subgroups of patients who are triple positive for PD-L1/CD8/IFN-γ, as shown in Example 4 below, allowed some subsets of patients to be identified with a possible response to anti-PD-1 antibody treatment. For example, many patients with MI-TN breast cancer (immunomodulatory, triple negative cancer) are triple positive for PD-L1/CD8/IFN-γ. MI-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 refers to cancers that do not express the estrogen receptor (ER), progesterone receptor (PR), and HER2/Neu. These cancers are difficult to treat because they usually do not respond to drugs that target ER, PR, and HER2/Neu. Triple-negative breast cancer can be further subdivided into different classes, one of which is immunomodulatory cancer. As described in Lehmann et al., IM-TN breast cancer is enriched in factors involved in immune cell processes, such as one or more of immune cell signaling (e.g., Th1/Th2 pathway, NK cell pathway, B cell signaling pathway). receptor, DC pathway, and T-cell receptor signaling), cytokine signaling (eg, cytokine pathway, IL-12 pathway, and IL-7 pathway), antigen processing and presentation, signaling via central immune signaling pathways (eg, NFKB signaling, TNF and JAK/STAT), genes involved in T cell function, immune transcription, interferon (IFN) release, and antigen processing. Thus, in some embodiments, the cancer being treated is a cancer that is or is determined to be positive for one or more breast cancer MI-TN markers, e.g., a factor that stimulates one or more of 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), processing and presentation antigen, signaling through central immune signaling pathways (eg, NFKB, TNF, and JAK/STAT signaling), genes involved in T cell function, immune transcription, interferon (IFN) release, and antigen processing.

As another example, the present application shows that a subset of high MSI (microsatellite instability) colon cancer patients are also triple positive for PD-L1/CD8/IFN-γ. Thus, in some embodiments, an anti-PD-1 antibody, e.g., an anti-PD-1 antibody of the invention (optionally in combination with one or more immunomodulators, such as an anti-LAG-3 antibody, an anti-TIM-3 antibody, or an anti-PD- L1, and one or more antitumor agents, for example, the antitumor agent described in Table 7 or in the publication of Table 7) is administered to a patient who has or is diagnosed with high MSI colon cancer, resulting in cancer treatment. In some embodiments, a high MSI cell is a cell that has a higher MSI level than a reference value or a value in a control cell, eg, a non-cancerous cell from the same type of tissue as the cancer.

As another example, the present application shows that a subset of gastric cancer patients with high MSI and/or who are EBV+ are also triple positive for PD-Ll/CD8/IFN-γ. Thus, in some embodiments, an anti-PD-1 antibody, such as an anti-PD-1 antibody of the invention (optionally in combination with one or more immunomodulators such as an anti-LAG-3 antibody, an anti-TIM-3 antibody, or an anti-PD- L1, and one or more antitumor agents, for example, the antitumor agent listed in Table 7 or in the publication from Table 7) is administered to a patient who has or is diagnosed with gastric cancer with high MSI and/or EBV+, resulting in cancer treatment . In some embodiments, a high MSI cell is a cell that has a higher MSI level than a reference value or a value in a control cell, eg, a non-cancerous cell from the same type of tissue as the cancer.

In addition, the present application discloses methods for testing cancer for PD-L1 and then treating cancer with an antibody to PD-1. As described in Example 5 below, a cancer sample can be tested for PD-L1 protein levels or mRNA levels. A sample containing a higher level of PD-L1 (protein or mRNA) than a reference value or a value in a control cell (eg, a non-cancerous cell) may be classified as PD-L1 positive. Thus, in some embodiments, an anti-PD-1 antibody, e.g., an anti-PD-1 antibody as described herein (optionally in combination with one or more antitumor agents) is administered to a patient who has or has been determined to be PD-L1 positive. cancer. The cancer may be, for example, non-small cell adenocarcinoma (ASA) of the lung (NSCLC), NSCLC squamous cell carcinoma (SCC), or hepatocellular carcinoma (HCC).

In some embodiments, the present methods comprise the use of an anti-PD-1 antibody, e.g., an anti-PD-1 antibody, as described herein, e.g., as monotherapy, for the treatment of cancer that is (or is determined to be) PD-L1 positive. . In some embodiments, the cancer is colon and rectal cancer (e.g., MSI-H), gastric cancer (e.g., MSI-H and/or EBV+), NPC, cervical cancer, breast cancer (e.g., TH breast cancer ) and ovarian cancer. In some embodiments, the cancer is NSCLC, melanoma, or HNSCC. In some embodiments, the anti-PD-1 antibody is administered at a dose of, for example, 1, 3, 10, or 20 mg/kg. In other embodiments, the anti-PD-1 antibody molecule is administered at a dose of, for example, 200, 250, 300, 350, 400, 450, or 500 mg. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of 250 mg to 450 mg, such as 250 mg, 300 mg, 350 mg, 400 mg, or 450 mg.

Based on, for example, Example 4 in the present application, it was found that some gastric cancers that are triple positive for PD-Ll/CD8/IFN-γ are also positive for PIK3CA. Thus, in some embodiments, cancer can be treated with an anti-PD-1 antibody molecule (optionally in combination with one or more immunomodulators, e.g., an anti-LAG-3 antibody molecule, an anti-TIM-3 antibody molecule, or an anti-PD-L1 antibody molecule) and agent that inhibits PIK3CA. Examples of 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). "Targeting phosphoinositide 3-kinase: moving towards therapy". Biochimica et Biophysica Acta 1784(1):159-85.

Based on, for example, Example 4 herein, a CRC of, for example, a patient who has (or is identified as having) an MSI-H CRC can be treated with an anti-PD-1 antibody, optionally in combination with a therapeutic agent that targets one or more of LAG-3, RNF43 and BRAF. By way of example, these types of cancer can be treated with an anti-PD-1 antibody, optionally in combination with one or more therapeutic agents that target one or more of LAG-3, PD-1, RNF43, and BRAF. In some embodiments, one or more therapeutic agents include immunomodulators, such as an anti-LAG-3 antibody molecule, and an antitumor agent described in Table 7 or in the publication listed in Table 7. LAG-3 inhibitors, such as antibodies, are described herein. application. RNF43 can be inhibited, e.g., by an antibody, a small molecule (e.g., 2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridine -2-yl)acetamide (Compound A28)), siRNA or Rspo ligand or derivatives thereof. BRAF inhibitors (eg vemurafenib or dabrafenib) are described in this application.

Based on, for example, Example 4 herein, a patient who has (or is identified as having) squamous cell lung cancer can be treated with an anti-PD-1 antibody molecule in combination with a therapeutic agent that targets LAG-3, for example, a molecule anti-LAG-3 antibodies, and optionally one or more antitumor agents, such as the antitumor agent described in Table 7 or in the publication in Table 7.

In some embodiments, a subject who has (or is identified as having) squamous cell lung cancer can be treated with an anti-PD-1 antibody, optionally in combination with a therapeutic agent that targets TIM-3, such as an anti-TIM-3 antibody. TIM-3 inhibitors, such as antibodies, are described in this application.

Based on, for example, Example 4 herein, a patient who has (or is identified as having) thyroid cancer (e.g., anaplastic thyroid carcinoma) can be treated with an anti-PD-1 antibody molecule, optionally in combination with a therapeutic agent that targets BRAF, optionally in combination with one or more immunomodulators, for example, an anti-LAG-3 antibody molecule, an anti-TIM-3 antibody molecule, and an anti-PD-L1 antibody molecule. BRAF inhibitors (eg, vemurafenib or dabrafenib) are described in this application, for example, in Table 7 and the publications listed in Table 7.

In some embodiments, the methods of treatment herein may be used to treat a patient who has (or is identified as having) a cancer associated with an infection, such as a viral or bacterial infection. Examples of cancer types include cervical cancer, rectal cancer, HPV-associated head and neck squamous cell carcinoma, HPV-associated esophageal papillomas, HHV6-associated lymphomas, EBV-associated lymphomas (including Burkitt's lymphoma), gastric MALT lymphoma, other associated MALT lymphoma infection, HCC, and Kaposi's sarcoma.

In other embodiments, the cancer is hemoblastosis or cancer, including, but not limited to, leukemia or lymphoma. For example, an anti-PD-1 antibody molecule can be used to treat types of cancers and malignancies including, but not limited to, for example, acute leukemias, including, but not limited to, B-cell acute lymphocytic leukemia ("B-ALL"). , T-cell acute lymphocytic leukemia ("T-ALL"), acute lymphocytic leukemia (ALL); one or more chronic leukemias, including, without limitation, for example, chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL); additional hematologic or hematologic disorders including, but not limited to, B-cell prolymphocytic leukemia, plasmacytoid dendritic cell blast tumor, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small-cell or large-cell follicular lymphoma, malignant lymphoproliferative conditions , MALT lymphoma, mantle cell lymphoma, marginal cell lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasia, Waldenström's macroglobulinemia and "preleukemia", which are a heterogeneous group of hematological conditions combined inefficient production (or dysplasia) of myeloid blood cells, and the like.

In one embodiment, the cancer is selected from lung cancer (e.g., non-small cell lung cancer (NSCLC) (e.g., squamous and/or non-squamous histology NSCLC, or adenocarcinoma NSCLC)), melanoma (e.g., advanced melanoma), kidney cancer (e.g., renal cell carcinoma, eg, clear cell renal cell carcinoma), liver cancer, myeloma (eg, multiple myeloma), prostate cancer, breast cancer (eg, breast cancer that does not express one, two, or all of the estrogen receptor, progesterone receptor or HER2/Neu e.g. triple negative breast cancer), colon and rectal 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 disease (eg, post-transplant lymphoproliferative disease) or hemoblastosis, T-cell mfoma, non-Hodgkin's lymphoma, or leukemia (eg, myeloid leukemia).

In another embodiment, the cancer is selected from carcinoma (eg, advanced or metastatic carcinoma), melanoma, or lung carcinoma, eg, non-small cell lung carcinoma.

In one embodiment, the cancer is lung cancer, such as non-small cell lung cancer.

In another embodiment, the cancer is a hepatocarcinoma, eg, advanced hepatocarcinoma, with or without a viral infection, eg, chronic viral hepatitis.

In another embodiment, the cancer is prostate cancer, eg advanced prostate cancer.

In yet another embodiment, the cancer is a myeloma, such as multiple myeloma.

In yet another embodiment, the cancer is kidney cancer, eg, renal cell carcinoma (RCC) (eg, metastatic RCC, non-clear cell renal cell carcinoma (nccRCC), or clear cell renal cell carcinoma).

In one embodiment, the cancer is melanoma, such as widespread melanoma. In one embodiment, the cancer is an advanced or inoperable melanoma that has not responded to other therapies. In other embodiments, the cancer is a BRAF-mutated melanoma (eg, a BRAF V600 mutation). In some embodiments, cancer (eg, melanoma) is present in a subject having an elevated serum LDH level relative to a reference serum LDH level. In other embodiments, the anti-PD-1 antibody molecule is administered following treatment with an anti-CTLA-4 antibody (eg, ipilimumab), with or without a BRAF inhibitor (eg, vemurafenib or dabrafenib). In one embodiment, the anti-PD-1 antibody molecule is administered with a BRAF inhibitor. For example, an anti-PD-1 antibody molecule may be administered at a dose of 200 mg to 600 mg (eg, 400 mg dose), such as once every 4 weeks or once every 8 weeks, such as intravenously, and a BRAF inhibitor or dabrafenib may be administered at a dose of 50 mg to 250 mg (eg 150 mg), twice a day, eg orally. In one embodiment, the combination further comprises a MEK inhibitor or trametinib.

In one embodiment, the cancer is colon and rectal cancer (CRC). Colon and rectal cancer is the third most common cancer in the world, with approximately 1.4 million people diagnosed in 2012, and the fourth most common cause of cancer death, with 694,000 deaths (World Cancer Report 2014). Treatment outcomes in CRC patients are associated with immune infiltrate in tumors, suggesting that therapies that stimulate an immune response may be effective in CRC (Fridman et al. 2011 Cancer Res. p. 5601-5). Without being limited by theory, it is believed that in some embodiments, combination therapy may be more effective in treating colon and rectal cancer, for example, except for the mismatch repair deficient subgroup when monotherapies are less effective (Kroemer et al. (2015) OncoImmunology 4:7, e1058597-1-3). For example, the combination of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) and a second therapeutic agent described herein may increase the percentage of patients with a response and/or duration of response compared to monotherapy (eg, a PD-1 inhibitor alone). or only the second therapeutic agent alone).

In another embodiment, the cancer is lung cancer, such as non-small cell lung cancer (NSCLC). Lung cancer is the most common cancer in the world, with approximately 1.8 million new cases in 2012, and the most common cause of cancer death, with 1.6 million deaths (World Cancer Report 2014). Of these cases, approximately 85% are non-small cell lung cancer (NSCLC). Inhibitors of the PD-1/PD-L1 interaction are used to treat NSCLC. The percentage of patients responding to these agents in a continuum is approximately 20% after first-line therapy fails (Gettinger and Herbst (2014) Cancer J. p. 281-9; Brahmer et al. (2012) N. Engl. J. Med. pp. 123-35 Topalian et al (2012) N. Engl J. Med pp 2443-54 Herbst et al (2014) Nature p 563-7 Borghaei et al (2015) N. Engl. J. Med. pp. 1627-39). In some embodiments, selection for PD-L1 expression in tumors may increase response to PD-1/PD-L1 inhibitors. Without being limited by theory, it is believed that in some embodiments, combination therapy may be more effective in the treatment of lung cancer (eg, NSCLC) compared to monotherapies. For example, the combination of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) and a second therapeutic agent described herein can increase the percentage of patients with a response and/or the duration of the response (e.g., extending activity, deepening the response, and/or obtaining longer responses) compared to monotherapy (for example, only a PD-1 inhibitor or only a second therapeutic agent alone).

Breast cancer is the second most common cancer in the world, with approximately 1.7 million new cases in 2012, and the fifth leading cause of cancer death, with approximately 521,000 deaths (World Cancer Report). Of these cases, approximately 15% are triple negative tumors that do not express the estrogen receptor (ER), progesterone receptor (PR), or HER2. As a result, targeted therapies that are available for patients with other subtypes of breast cancer usually cannot be effectively applied in these patients. Triple-negative breast cancer (TNBC) is an aggressive disease and outcomes after treatment are poor (Foulkes et al. (2010) N. Engl. J. Med. p. 1938-48). Recent data indicate that PD-1 inhibitors are active in some patients, with a response noted in five of 27 pembrolizumab-treated patients with metastatic TNBC in the Keynote-012 study (Buisseret et al. (2015) Annals of Oncology 26(suppl_3 ): pp. 6-9). Without being limited by theory, it is believed that in some embodiments, combination therapy may be more effective in treating breast cancer (eg, TNBC) compared to monotherapies. For example, the combination of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) and a second therapeutic agent described herein may increase the percentage of patients who respond and/or the duration of response (e.g., increase the percentage of patients who respond to treatment). ) compared to monotherapy (eg, PD-1 inhibitor alone or second therapeutic agent alone).

The methods and compositions disclosed in this application can be used to treat metastatic lesions associated with the above forms of cancer.

Combination Therapy

Non-limiting examples of combinations and uses of anti-PD-1 antibody molecules are disclosed in US Patent Application Publication 2015/0210769 (USSN 14/604,415) entitled "Antibody Molecules to PD-1 and Uses Thereof", incorporated by reference in its entirety.

In some embodiments, the combination comprises an anti-PD-1 antibody molecule in combination with a modulator of a co-stimulatory molecule or an inhibitory molecule, such as a co-inhibitory ligand or receptor.

In one embodiment, an anti-PD-1 antibody molecule is administered in combination with a modulator, such as an agonist, of a costimulatory molecule. In one embodiment, the costimulatory molecule agonist is selected from an agonist (e.g., agonist antibody or antigen-binding fragment thereof, or soluble fusion construct) OX40, CD2, CD27, CD, 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.

In another embodiment, an anti-PD-1 antibody molecule is used in combination with a co-stimulatory molecule, eg, a positive signal-associated agonist, which includes the co-stimulatory domain of CD28, CD27, ICOS, and GITR.

In one embodiment, an anti-PD-1 antibody molecule is administered in combination with an inhibitor molecule that inhibits an immune checkpoint molecule. Those skilled in the art will recognize that the term "immune checkpoints" refers to a group of molecules on the cell surface of CD4 and CD8 T cells. These molecules can effectively serve as a "brake" to downmodulate or inhibit the antitumor immune response. Immune checkpoint molecules include, but are not limited to, programmed death protein 1 (PD-1), cytotoxic T lymphocyte antigen 4 (CTLA-4), B7H1, B7H4, OX-40, CD137, CD40, LAG-3, and TIM- 3, which directly inhibit immune cells. Immunotherapeutic agents that may act as immune checkpoint inhibitors useful in the methods of the present invention include, but are not limited to, inhibitors of PD-L1, PD-L2, CTLA-4, TIM-3, LAG-3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, CEACAM (eg CEACAM-1 and/or CEACAM-5) and/or TGF-beta.

In one embodiment, the inhibitor is a soluble ligand (eg, CTLA-4-Ig or TIM-3-Ig) or an antibody or antibody fragment that binds to PD-L1, PD-L2, or CTLA-4. For example, an anti-PD-1 antibody molecule can be administered in combination with an anti-CTLA-4 antibody, e.g., ipilimumab, e.g., for the treatment of cancer (e.g., a cancer selected from: melanoma, e.g., metastatic melanoma; lung cancer, e.g., non-small cell carcinoma lung; or prostate cancer). Examples of anti-CTLA-4 antibodies include tremelimumab (an IgG2 monoclonal antibody manufactured by Pfizer, formerly known as ticilimumab, CP-675,206); and ipilimumab (an anti-CTLA-4 antibody also known as MDX-010, CAS number 477202-00-9). In one embodiment, the PD-1 antibody molecule is administered post-treatment, eg, after melanoma treatment, with an anti-CTLA-4 antibody (eg, ipilimumab), with or without a BRAF inhibitor (eg, vemurafenib or dabrafenib). Exemplary doses that may be used include an anti-PD-1 antibody molecule dose of approximately 1-10 mg/kg, such as 3 mg/kg, or 200-500 mg, such as 300 mg or 400 mg), and an anti-CTLA-4 antibody dose. e.g. ipilimumab, approximately 3 mg/kg.

Immune inhibitory molecules, eg PD-1 and LAG-3, can regulate, eg regulate synergistically, T cell function, causing immune escape of tumors. In another embodiment, an anti-PD-1 antibody molecule is administered in combination with an anti-LAG-3 antibody or antigen-binding fragment thereof. In another embodiment, an anti-PD-1 antibody molecule is administered in combination with an anti-TIM-3 antibody or antigen-binding fragment thereof. In other embodiments, an anti-PD-1 antibody molecule is administered in combination with an anti-LAG-3 antibody and an anti-TIM-3 antibody, or antigen-binding fragments thereof. The combination of antibodies specified in this application can be administered separately, for example, as separate antibodies, or as a linked molecule, for example, a bispecific or trispecific antibody molecule. In another embodiment, an anti-PD-1 antibody molecule is administered in combination with a CEACAM inhibitor (eg, a CEACAM-1, CEACAM-3 and/or CEACAM-5 inhibitor), for example, an anti-CEACAM antibody molecule. In another embodiment, an anti-PD-1 antibody molecule is administered in combination with a CEACAM-1 inhibitor, eg, an anti-CEACAM-1 antibody molecule. In another embodiment, an anti-PD-1 antibody molecule is administered in combination with a CEACAM-5 inhibitor, eg, an anti-CEACAM-5 antibody molecule. In one embodiment, a bispecific antibody is administered that comprises an anti-PD-1 antibody molecule and an anti-TIM-3 or anti-LAG-3 antibody, or an antigen-binding fragment thereof. In some embodiments, the combination of antibodies specified in this application is used to treat cancer, for example, cancer described in this application (eg, solid tumor). The effectiveness of the above combinations can be tested in animal models known in the art. For example, animal models for testing synergistic effect against PD-1 and against LAG-3 are described, for example, in Woo et al. (2012) Cancer Res. 72(4):917-27.

In one embodiment, the CEACAM inhibitor (eg, CEACAM-1 and/or CEACAM-5) is an anti-CEACAM antibody molecule. Without being limited by theory, CEACAM-1 has been described as a ligand and partner of TIM-3 (see, for example, WO 2014/022332). A synergistic effect in vivo of a combination of anti-TIM-3 and anti-CEACAM-1 antibodies has been found in xenograft tumor models (see, for example, WO 2014/022332). It is believed that tumors use CEACAM-1 or CEACAM-5 to inhibit the immune system, as described, for example, in Markel et al. J Immunol. 2002 Mar 15; 168(6):2803-10; Markel et al. J Immunol. 2006 Nov 1; 177(9):6062-71; Markel et al. Immunology. February 2009; 126(2):186-200; Markel et al. Cancer Immunol Immunother. February 2010; 59(2):215-30; Ortenberg et al. Mol Cancer Ther. Jun 2012; 11(6):1300-10; Stern et al. J Immunol. 2005 Jun 1; 174(11):6692-701; Zheng et al. PLOS One. 2010 Sep 2; 5(9). pii:e12529. Thus, CEACAM inhibitors can be used with other immunomodulators described in this application (for example, with inhibitors against PD-1 or against TIM-3) to enhance the immune response against cancer, for example, melanoma, lung cancer (for example, NSCLC), cancer bladder, colon or ovary, or other types of cancer described in this application. In one embodiment, the CEACAM inhibitor is an anti-CEACAM-1 antibody as described in WO 2010/125571, WO 2013/82366 and WO 2014/022332, e.g., a 34B1, 26H7, and 5F4 monoclonal antibody, or a recombinant form thereof, as described, for example, in US 2004/0047858, US 7,132,255 and WO 99/52552. In other embodiments, the anti-CEACAM antibody is an anti-CEACAM-1 antibody molecule and/or an anti-CEACAM-5 antibody molecule as described, for example, in WO 2010/125571, WO 2013/054331 and US 2014/0271618.

In some embodiments, PD-1 and LAG-3 immune inhibitory molecules (eg, antibody molecules) are administered in combination with each other, eg, for the treatment of cancer. In some embodiments, the patient is a patient who has experienced progression (e.g., marked tumor growth) during therapy with a PD-1 inhibitor (e.g., an antibody molecule described herein) and/or a PD-L1 inhibitor (e.g., an antibody molecule) . In some embodiments, therapy with a PD-1 antibody molecule and/or a PD-L1 antibody molecule is continued and an immune inhibitory molecule (eg, antibody) is further included in the LAG-3 therapy.

In some embodiments, PD-1 and TIM-3 immune inhibitory molecules (eg, antibody molecules) are administered in combination with each other, for example, for the treatment of cancer. In some embodiments, the patient is a patient who has experienced progression (e.g., marked tumor growth) during therapy with a PD-1 inhibitor (e.g., an antibody molecule described herein) and/or a PD-L1 inhibitor (e.g., an antibody molecule ). In some embodiments, therapy with a PD-1 antibody molecule and/or a PD-L1 antibody molecule is continued and an immune inhibitory molecule (eg, antibody) is additionally included in the TIM-3 therapy.

In other embodiments, the anti-PD-1 antibody molecule is administered in combination with a cytokine, such as interleukin-21, interleukin-2, interleukin-12, or interleukin-15. In some embodiments, the combination of an anti-PD-1 antibody molecule and a cytokine described herein is used to treat cancer, eg, the cancer described herein (eg, solid tumor or melanoma).

Examples of immunomodulators that can be used in combination with anti-PD-1 antibody molecules include, but are not limited to, for example, afutuzumab (manufactured by Roche®); pegfilgrastim (Neulastim®); lenalidomide (CC-5013, Revlimid®); thalidomide (Thalomid®), actimide (CC4047); and cytokines, for example, IL-21 or IRX-2 (mixture of human cytokines, including interleukin 1, interleukin 2 and interferon γ, CAS 951209-71-5, manufactured by IRX Therapeutics).

In other embodiments, an anti-PD-1 antibody molecule is used in combination with an indolamine-pyrrole-2,3-dioxygenase (IDO) inhibitor (e.g., INCB24360) in a subject with advanced or metastatic cancer (e.g., a patient with metastatic and recurrent NMLC cancer). ).

In other embodiments, the combinations disclosed herein, e.g., a combination comprising an anti-PD-1 antibody molecule, is administered to a subject in combination with (e.g., before, concurrently, or after) one or more of the following: bone marrow transplantation, anti-PD-1 ablative therapy. T cells using chemotherapeutic agents such as fludarabine, external beam radiation therapy (XRT), cyclophosphamide, and/or antibodies such as OKT3 or Campas. In one embodiment, combinations, eg, combinations comprising an anti-PD-1 antibody molecule, are administered following anti-B cell ablative therapy using agents that interact with CD20, eg, Rituxan. For example, in one embodiment, subjects may be treated with standard high dose chemotherapy followed by peripheral blood stem cell transplantation. In some embodiments, after transplantation, subjects receive combinations. In an additional embodiment, the combinations are administered before or after surgery.

Another example of a combination is an anti-PD-1 antibody in combination with dacarbazine for the treatment of melanoma. Other combination therapies that may result in synergy with PD-1 blockade through cell apoptosis include radiation, surgery, and hormonal deprivation. Each of these protocols creates a source of tumor antigen in the host organism. Angiogenesis inhibitors can also be combined with PD-1 blockade. Inhibition of angiogenesis results in tumor cell death, which can deliver tumor antigen in the host antigen presentation pathway.

Combinations that include blocking PD-1 antibodies can also be used in combination with bispecific antibodies. Bispecific antibodies can be used to target two separate antigens. For example, anti-Fc receptor/tumor antigen bispecific antibodies (eg, Her-2/Neu) have been used to direct macrophages to tumor sites. This direction can more effectively activate tumor-specific responses. The T cell arm of such responses will be enhanced by the application of PD-1 blockade. Alternatively, the antigen can be delivered directly to the DC using bispecific antibodies that bind to the tumor antigen and a specific dendritic cell surface marker.

Tumors evade immune surveillance through a variety of mechanisms. Many of these mechanisms can be overcome by deactivating proteins that are expressed by tumors and that are immunosuppressive. These include, among others, TGF-beta (Kehrl, J. et al. (1986) J. Exp. Med. 163: 1037-1050), IL-10 (Howard, M. & O'Garra, A. (1992 ) Immunology Today 13: 198-200) and Fas ligand (Hahne, M. et al. (1996) Science 274: 1363-1365). The compositions disclosed herein, which include anti-PD-1 antibodies, can antagonize the action of an immunosuppressant and promote the induction of immune responses against a tumor in the host.

Other antibodies that can be used to activate an immune response in a host can be used in combination with an anti-PD-1 antibody molecule as described herein. They include molecules on the surface of dendritic cells (DCs) that activate DC function and antigen presentation. Anti-CD40 antibodies can effectively replace T-helper cell activity (Ridge, J. et al. (1998) Nature 393:474-478) and can be used in combination with anti-PD-1 antibodies (Ito, N. et al. (2000 ) Immunobiology 201(5)527-40). Antibodies against costimulatory T cell molecules such as CTLA-4 (e.g. US Patent 5,811,097), OX-40 (Weinberg, A. et al. (2000) Immunol 164:2160-2169), 4-1BB (Melero, I et al (1997) Nature Medicine 3:682-685 (1997) and ICOS (Hutloff, A. et al. (1999) Nature 397:262-266) may also provide increased levels of T cell activation.

Additional examples of standard therapy are described in the section titled "Combination Therapies" in US Patent Application Publication 2015/0210769 (USSN 14/604,415) entitled "Antibody Molecules to PD-1 and Uses Thereof", incorporated by reference in its entirety, and below.

In all the methods described herein, PD-1 blockade can be combined with other forms of immunotherapy such as cytokine treatment (eg, interferons, GM-CSF, G-CSF, IL-2, IL-21) or bispecific antibody therapy. , which provides for increased presentation of tumor antigens (see, for example, Holliger (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak (1994) Structure 2:1121-1123).

Methods for introducing antibody molecules are known in the art and are described below. Suitable dosages of molecules used will depend on the age and body weight of the subject, as well as the particular drug used. Doses and treatment regimens using an anti-PD-1 antibody molecule can be determined by those skilled in the art. In some embodiments, the anti-PD-1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose of about 1-30 mg/kg, e.g., about 5-25 mg/kg, about 10-20 mg/kg, about 1 -5 mg/kg or approximately 3 mg/kg. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 1 mg/kg, about 3 mg/kg, about 5 mg/kg, about 10 mg/kg, about 20 mg/kg, about 30 mg/kg, or about 40 mg/kg. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 1-3 mg/kg, or about 3-10 mg/kg. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 0.5-2, 2-4, 2-5, 5-15, or 5-20 mg/kg. The administration schedule may vary, for example, from once a week to once every 2, 3 or 4 weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 10-20 mg/kg biweekly. In another embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 1 mg/kg once every two weeks, about 3 mg/kg once every two weeks, 10 mg/kg once every two weeks, 3 mg/kg once every every four weeks or 5 mg/kg once every four weeks.

In other embodiments, the anti-PD-1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose (e.g., fixed dose) of from about 200 mg to 500 mg, for example, from about 250 mg to 450 mg, from about 300 mg up to 400 mg, about 250 mg to 350 mg, about 350 mg to 450 mg or about 300 mg or about 400 mg. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 250-450 mg or about 300-400 mg. In some embodiments, the anti-PD-1 antibody molecule is administered at a dose of about 200-300 mg, 250-350 mg, 300-400 mg, 350-450 mg, or 400-500 mg. The administration schedule may vary, for example, from once a week to once every 2, 3, 4, 5 or 6 weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg to 400 mg once every three or once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every three weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every four weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every three weeks.

The antibody molecules can be used in unconjugated forms or as a conjugate with a second agent, such as a cytotoxic agent, a radioisotope, or a protein, such as a protein toxin or a viral protein. This method includes: administering the antibody molecule, alone or as a conjugate to a cytotoxic agent, to a subject in need of such treatment. Antibody molecules can be used to deliver various therapeutic agents, e.g., a cytotoxic molecule, e.g., a therapeutic drug, a radioisotope, a molecule of plant, fungal, or bacterial origin, or biological proteins (e.g., protein toxins) or particles (e.g., recombinant viral particles, e.g. via the viral envelope protein), or mixtures thereof.

Additional Combination Therapies

The combinations disclosed herein, eg a combination comprising PD-1 blocking agents, can also be combined with standard cancer treatment, eg chemotherapeutic regimens. In such cases, the dose of chemotherapeutic agent administered can be reduced (Mokyr, M. et al. (1998) Cancer Research 58:5301-5304). In some embodiments, the methods and compositions described herein are used in combination with one or more of other antibody molecules, chemotherapy, other anticancer therapy (e.g., targeted anticancer agents or oncolytic agents), cytotoxic agents, immunotherapeutic agents (e.g., cytokines), surgery and/or radiotherapy procedures. Examples of cytotoxic agents that can be administered in combination include antimicrotubulin agents, topoisomerase inhibitors, antimetabolites, mitosis inhibitors, alkylating agents, anthracyclines, vinca alkaloids, intercalating agents, agents capable of disrupting signal transduction pathways, agents that induce apoptosis, inhibitors of the proteasome and radiation therapy (for example, local or total body irradiation).

Alternatively, or in combination with the above combinations, the methods and compositions described herein may be used in combination with one or more of the following: an immunomodulator (eg, an activator of a costimulatory molecule or an inhibitor of an inhibitory molecule), a vaccine, eg, a therapeutic cancer vaccine , or other forms of cellular immunotherapy.

Examples of combinations, for example combinations comprising anti-PD-1 antibody molecules and a standard cancer treatment, are disclosed in US Patent Application Publication 2015/0210769 (USSN 14/604,415) entitled "Antibody Molecules to PD-1 and Uses Thereof", fully incorporated by reference, including, without limitation, alkylating agents, anthracyclines, vinca alkaloids, proteasome inhibitors, and tyrosine kinase inhibitors (eg, receptor tyrosine kinase (RTK) inhibitor).

An exemplary tyrosine kinase inhibitor includes, but is not limited to, an epidermal growth factor (EGF) pathway inhibitor (e.g., an epidermal growth factor receptor (EGFR) inhibitor), a vascular endothelial growth factor (VEGF) pathway inhibitor (e.g., a vascular endothelial growth factor receptor (VEGFR) inhibitor). ) (eg VEGFR-1 inhibitor, VEGFR-2 inhibitor, VEGFR-3 inhibitor)), platelet growth factor (PDGF) pathway inhibitor (eg platelet growth factor receptor (PDGFR) inhibitor (eg PDGFR-β inhibitor)), RAF-1 inhibitor, KIT inhibitor and RET inhibitor. Selected tyrosine kinase inhibitors are selected from sunitinib, erlotinib, gefitinib, or sorafenib. Sorafenib is also known as 4-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-N-methyl-pyridine-2-carboxamide. In some embodiments, the implementation of the combination includes sorafenib, for example, for the treatment of cancer described in this application, for example, liver cancer (eg, hepatocellular carcinoma).

In some embodiments, the combinations include vascular endothelial growth factor (VEGF) receptor inhibitors, such as a VEGFR inhibitor, as described herein.

In some embodiments, the combination includes a PI3K inhibitor, such as a PI3K inhibitor, as described herein.

In some embodiments, the combination includes an mTOR inhibitor, such as an mTOR inhibitor, as described herein.

In some embodiments, the combination includes a BRAF inhibitor, such as GSK2118436, RG7204, PLX4032, GDC-0879, PLX4720, and sorafenib tosylate (Bay 43-9006). In some embodiments, the combination includes a RAF inhibitor, such as debrafenib or N-{3-[5-(2-aminopyrimidin-4-yl)-2-tert-butyl-1,3-thiazol-4-yl]-2- fluorophenyl}-2,6-difluorobenzenesulfonamide.

In some embodiments, the combination includes a MEK inhibitor. In some embodiments, the cancer being treated with the combination is selected from melanoma, colon and rectal cancer, non-small cell lung cancer, ovarian cancer, breast cancer (e.g., triple negative breast cancer (TNBC)), prostate cancer, pancreatic cancer. , hemoblastosis, or renal cell carcinoma. In some embodiments, the cancer comprises a BRAF mutation (eg, a BRAF V600E mutation), wild-type BRAF, wild-type KRAS, or an activating KRAS mutation. Cancer can be early, intermediate, or advanced. In one embodiment, the cancer being treated with the combination is unresectable or metastatic melanoma, eg, having a BRAFV600 mutation. In some embodiments, the cancer is selected from colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS CRC), lung cancer (e.g., non-small cell lung cancer), or breast cancer (e.g., triple negative lung cancer (TNBC) ).

Any MEK inhibitor may be used in combination, including, but not limited to, ARRY-142886, G02442104 (also known as GSK1120212), RDEA436, RDEA119/BAY 869766, AS703026, G00039805 (also known as AZD-6244 or selumetinib), BIX 02188, BIX 02189, CI-1040 (PD-184352), 032590PD-1, PD98059, U0126, GDC-0973 ([3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl][3- hydroxy-3-(25)-2-piperidinyl-1-azetidinyl]methanone), G-38963, G02443714 (also known as AS703206), or a pharmaceutically acceptable salt or solvate thereof. Additional examples of MEK inhibitors are disclosed in WO 2013/019906, WO 03/077914, WO 2005/121142, WO 2007/04415, WO 2008/024725 and WO 2009/085983, the contents of which are hereby incorporated by reference.

In some embodiments, the MEK inhibitor is trametinib or N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3, 4,6,7-tetrahydro-pyrido[4,3-d]pyrimidin-1(2H)-yl}phenyl)acetamide.

In some embodiments, the MEK inhibitor or trametinib is a reversible and highly selective allosteric inhibitor of MEK1 and MEK2. MEK proteins are important components of the MAPK pathway, which is typically overactivated in tumor cells. Oncogenic mutations in BRAF (a representative of RAF kinases) and RAS signal through MEK1 or MEK2. Without being limited by theory, it is believed that in some embodiments, an MEK inhibitor, such as trametinib, can increase CD4+ and CD8+ tumor infiltrating lymphocytes (TILs), and/or increase the efficacy of a PD-1 inhibitor (Liu et al. (2014) Clin. Cancer Res. p. 462-8).

In some embodiments, the combination described herein includes a MEK inhibitor or trametinib, and an inhibitor of an immune checkpoint molecule, such as a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule).

MAPK pathway signaling is important for tumor growth (Downward (2003) Nat. Rev. Cancer. p. 11-22) and trametinib is effective alone and in combination with dabrafenib for the treatment of BRAFV600 mutant melanoma. Trametinib produces MEK inhibition in tumor cells at exposures achievable at an approved dose of 2 mg po once daily (QD) (Infante et al. (2012) Lancet Oncol. p. 773-81). Preclinical data from a syngeneic mouse model of CT26 demonstrate that the combination of trametinib and an anti-PD-L1 antibody results in increased TIL, reduced tumor growth, and increased survival (Liu et al. (2015) Clin. Cancer Res. p. 1639 -51). The immunomodulatory effects of trametinib in combination with anti-checkpoint inhibitor antibodies were investigated in a syngeneic in vivo CT26 mouse model. For example, trametinib increases tumor infiltrating lymphocytes (TIL) and trametinib potentiates antitumor effects when treated with anti-PD-1 antibody, anti-PD-L1 antibody, and anti-CTLA-4 antibody. Using this model, mice were treated with control, trametinib, anti-PD-1 antibody, or a combination of trametinib and anti-PD-1 antibody for seven days, after which tumors were harvested. Treatment with trametinib or trametinib and anti-PD-1 antibody for seven days resulted in significant increases in CD4+ T cells in the tumor tissue. Only treatment with the combination of trametinib and anti-PD-1 antibody resulted in significant increases in CD8+ T cells. In addition, in the CT26 mouse model, the combination of trametinib and anti-PD-1 antibody was more effective in inhibiting tumor growth than either agent alone. The combination treatment also increased the duration of survival. The dosage regimen can be important. Tumor growth was inhibited by co-administration of trametinib and anti-PD-1 antibody, or when trametinib was first administered as a single agent for 7 days prior to combination treatment. Tumor growth was not inhibited if the antibody was first administered and then the combination therapy was used, indicating that the drug regimen may be important. In some embodiments, a MEK inhibitor or trametinib enhances or is used to enhance the antitumor effect of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule).

In some embodiments, a MEK inhibitor or trametinib and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), are administered at a dose and/or timing that, in combination, provides the desired antitumor activity.

In one embodiment, the MEK inhibitor or trametinib is administered at a dose of 0.2 mg to 5 mg, e.g., 0.3 mg to 4 mg, 0.4 mg to 3 mg, 0.5 mg to 2 mg, 1 mg to 1.5 mg, 1.5 mg to 2 mg, or 0.4 mg to 0.6 mg, for example, at a dose of 0.2 mg, 0.5 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg, 3 mg, 4 mg or 5 mg, for example, twice a day, once a day, once every other day, once every three days, or once a week.

In one embodiment, the MEK inhibitor or trametinib is administered at a dose of 0.4 mg to 0.6 mg (eg, 0.5 mg) once daily. In another embodiment, the MEK inhibitor or trametinib is administered at a dose of 1 mg to 3 mg (eg, 2 mg) once a day. In some embodiments, the MEK inhibitor or trametinib is administered orally. In one embodiment, the MEK inhibitor or trametinib is administered at a dose of 0.4 mg to 0.6 mg (e.g., 0.5 mg), e.g., once a day, e.g., orally, and a PD-1 inhibitor (e.g., , an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (eg 400 mg dose), eg once every 4 weeks, eg by intravenous infusion. In one embodiment, the MEK inhibitor or trametinib is administered at a dose of 1 mg to 3 mg (e.g., 2 mg), e.g., once a day, e.g., orally, and a PD-1 inhibitor (e.g., an anti-PD- 1) is administered at a dose of 300 mg to 500 mg (eg 400 mg dose), eg once every 4 weeks or once every 8 weeks, eg by intravenous infusion.

In one embodiment, a MEK inhibitor or trametinib is used in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) to treat melanoma (e.g., metastatic or inoperable melanoma), colon and rectal cancer (CRC), lung cancer ( for example, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple-negative breast cancer (TNBC)). In some embodiments, the melanoma has a BRAF mutation, such as a BRAF V600 mutation. In some embodiments, the combination further comprises a BRAF inhibitor, such as dabrafenib. In some embodiments, the combination is used to treat a subject having an elevated serum LDH level relative to a reference serum LDH level.

In some embodiments, the combination includes a JAK2 inhibitor such as CEP-701, INCB18424, CP-690550 (tazocitinib).

In some embodiments, the combination includes paclitaxel or a paclitaxel-based agent, such as Taxol®, a protein-bound paclitaxel (eg, Abraxane®).

Radiation therapy may be administered by one of more methods or a combination of methods, including, without limitation, external beam radiation therapy, internal beam radiation therapy, implantation radiation, stereotaxic radiosurgery, systemic radiation therapy, radiotherapy, and permanent or temporary interstitial brachytherapy.

The disclosed combinations may be used in combination with one or more existing cancer therapies, including, but not limited to: surgery, radiation therapy (e.g., external beam radiation therapy, which includes three-dimensional, conformal radiation therapy, in which the radiation field is indicated, local radiation (e.g. irradiation directed at a preselected target or organ) or focused irradiation). Focused radiation may be selected from the group consisting of stereotactic radiosurgery, fractionated stereotaxic radiosurgery, and intensity modulated radiation therapy. In focused irradiation, the radiation source can be selected from the group consisting of a particle beam (protons), cobalt-60 (photons) and a linear accelerator (X-ray), for example, as described in WO 2012/177624.

In some embodiments, the combinations include an anti-killer cell immunoglobulin-like receptor antibody (also referred to herein as an "anti-KIR antibody"), a pan-KIR antibody, an anti-NKG2D antibody, and an anti-MICA antibody. In some embodiments, the combination of an anti-PD-1 antibody molecule and an anti-KIR antibody, a pan-KIR antibody, or an anti-NKG2D antibody described herein is used to treat a cancer, e.g., the cancer described herein (e.g., a solid tumor, e.g. , advanced solid tumor).

In one embodiment, the combination includes cellular immunotherapy (eg, Provenge (eg, Cipuleucel)) and optionally in combination with cyclophosphamide. In some embodiments, a combination of an anti-PD-1 antibody molecule, Provenge, and/or cyclophosphamide is used to treat cancer, eg, the cancer described herein (eg, prostate cancer, eg, advanced prostate cancer).

In another embodiment, the combination comprises a vaccine, for example, a dendritic cell based renal cell carcinoma (DC-RCC) vaccine. In some embodiments, the combination of an anti-PD-1 antibody molecule and a DC-RCC vaccine is used to treat cancer, e.g., the cancer described herein (e.g., kidney cancer, e.g., metastatic renal cell carcinoma (RCC) or clear cell renal cell carcinoma). carcinomas (CCRCC)).

In yet another embodiment, an anti-PD-1 antibody molecule, alone or in combination with another immunomodulator (eg, an anti-LAG-3, anti-PD-L1, or anti-TIM-3 antibody molecule), is used in combination with chemotherapy and/or immunotherapy. For example, an anti-PD-1 antibody molecule can be used to treat myeloma, alone or in combination with one or more of: chemotherapy or other anticancer agents (e.g., thalidomide analogs, e.g., lenalidomide), anti-TIM-3 antibodies, tumor antigen sensitized dendritic cells, fusions (eg, electrofusions) of tumor cells and dendritic cells, or vaccination using an immunoglobulin idiotype produced by malignant plasma cells. In one embodiment, an anti-PD-1 antibody molecule is used in combination with an anti-TIM-3 antibody for the treatment of myeloma, eg, multiple myeloma.

In one embodiment, an anti-PD-1 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-L1, or anti-TIM-3 antibody molecule), is used in combination with chemotherapy to treat lung cancer, e.g. , non-small cell lung cancer. In one embodiment, an anti-PD-1 antibody molecule is used with platinum dual therapy for the treatment of lung cancer.

In yet another embodiment, an anti-PD-1 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-L1, or anti-TIM-3 antibody molecule), is used to treat kidney cancer, for example, renal cell carcinoma (RCC) (eg, clear cell renal cell carcinoma (CCRCC), non-clear cell renal cell carcinoma (nccRCC), or metastatic RCC). The anti-PD-1 antibody molecule can be used in combination with one or more of: an immune strategy (eg, interleukin-2 or interferon-α), a targeting agent (eg, a VEGF inhibitor such as an anti-VEGF monoclonal antibody), a VEGF tyrosine kinase inhibitor , such as sunitinib, sorafenib, axitinib and pazopanib, an RNAi inhibitor) or a downstream mediator inhibitor of the VEGF signaling pathway, such as the mammalian target of rapamycin (mTOR) inhibitor, such as everolimus and temsirolimus. In some embodiments, the combination includes sorafenib, eg, for the treatment of the cancer described herein, eg, liver cancer (eg, hepatocellular carcinoma).

An example of suitable therapeutic agents for use in combination with the anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-L1 or anti-TIM-3 antibody molecule), for treatment of pancreatic cancer includes, but is not limited to, a chemotherapeutic agent, such as paclitaxel or a paclitaxel-based agent (eg, a paclitaxel formulation such as Taxol, an albumin-stabilized nanoparticulate formulation of paclitaxel (eg, Abraxane) or a liposomal formulation of paclitaxel ), gemcitabine (eg, gemcitabine alone or in combination with AChP107-11), other chemotherapy agents such as oxaliplatin, 5-fluorouracil, capecitabine, rubitecan, epirubicin hydrochloride, NC-6004, cisplatin, docetaxel (eg, Taxotere), mitomycin C, ifosfamide; interferon; tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib, panitumumab, cetuximab, nimotuzumab); HER2/Neu receptor inhibitor (eg, trastuzumab); dual kinase inhibitor (eg, bosutinib, saracatinib, lapatinib, vandetanib); multikinase inhibitor (eg, sorafenib, sunitinib, XL184, pazopanib); VEGF inhibitor (eg, bevacizumab, AV-951, brivanib); radioimmunotherapy (eg, XR303), cancer vaccine (eg, GVAX, survivin peptide); COX-2 inhibitor (eg, celecoxib) ; IGF-1 receptor inhibitor (eg, AMG 479, MK-0646); mTOR inhibitor (eg, everolimus, temsirolimus); IL-6 inhibitor (eg, CNTO 328); cyclin-dependent kinase inhibitor (eg, P276-00, UCN-01); compound targeting altered energy metabolism (AEMD) (eg CPI-613); HDAC inhibitor (eg vorinostat); TRAIL receptor 2 (TR-2) agonist (eg conatumumab); MEK inhibitor ( e.g. AS703026, selumetinib, GSK1120212); dual inhibitor kinase Raf/MEK (eg R05126766); inhibitor of the Notch signaling pathway (for example, MK0752); monoclonal antibody-antibody fusion protein (eg, L19IL2); curcumin; an HSP90 inhibitor (eg tanespimycin, STA-9090); rIL-2, denileukin diftitox; a topoisomerase 1 inhibitor (eg, irinotecan, PEP02); a statin (eg simvastatin); a factor VIIa inhibitor (eg, PCI-27483); an AKT inhibitor (eg RX-0201); hypoxia-activated prodrug (eg, TH-302); metformin hydrochloride, gamma secretase inhibitor (eg RO4929097); a ribonucleotide reductase inhibitor (eg, 3-AP); immunotoxin (eg HuC242-DM4); a PARP inhibitor (eg KU-0059436, veliparib); a CTLA-4 inhibitor (eg, CP-675206, ipilimumab); AdV-tk therapy; a proteasome inhibitor (eg, bortezomib (Velcade), NPI-0052); thiazolidinedione (eg pioglitazone); NPC-1C; Aurora kinase inhibitor (eg R763/AS703569), CTGF inhibitor (eg FG-3019); siG12D-LODER; and radiation therapy (eg, tomotherapy, stereotactic radiation, proton therapy), surgery, and a combination thereof. In some embodiments, a combination of paclitaxel or a paclitaxel and gemcitabine agent with the anti-PD-1 antibody molecules described herein may be used.

An example of suitable therapeutic agents for use in combination with anti-PD-1 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-L1 or anti-TIM-3 antibody molecule), for the treatment of small cell lung cancer includes , without limitation, a chemotherapeutic agent, e.g., etoposide, carboplatin, cisplatin, oxaliplatin, irinotecan, topotecan, gemcitabine, SN-38 liposomal, bendamustine, temozolomide, belotecan, NK012, FR901228, flavopiridol); tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib, gefitinib, cetuximab, panitumumab); multikinase inhibitor (eg, sorafenib, sunitinib); VEGF inhibitor (eg, bevacizumab, vandetanib); cancer vaccine (eg, GVAX); Bcl- inhibitor 2 (eg, oblimersen sodium, ABT-263); proteasome inhibitor (eg, bortezomib (Velcade), NPI-0052), paclitaxel or paclitaxel-based agent; docetaxel; IGF-1 receptor inhibitor (eg, AMG 479); HGF inhibitor /SF (eg, AMG 102, MK-0646); chloroquine; Aurora kinase inhibitor (eg, MLN8237); radioimmunotherapy (eg, TF2); HSP90 inhibitor (eg, tanespimycin, STA-9090); mTOR inhibitor (eg, everolimus ); anti-Ep-CAM/CD3 bispecific antibody (eg, MT110); CK-2 inhibitor (eg, CX-4945); HDAC inhibitor (eg, belinostat); SMO antagonist (eg, BMS 833923); peptide tumor vaccine and radiation therapy (for example, radiation therapy with modulated intensity (IMRT), hypofractionated radiotherapy, hypoxia-guided radiotherapy), surgery, and combinations thereof.

An example of suitable therapeutic agents for use in combination with anti-PD-1 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-L1 or anti-TIM-3 antibody molecule), for the treatment of non-small cell lung cancer includes , without limitation, a chemotherapeutic agent, e.g., vinorelbine, cisplatin, docetaxel, pemetrexed disodium, etoposide, gemcitabine, carboplatin, liposomal SN-38, TLK286, temozolomide, topotecan, pemetrexed disodium, azacitidine, irinotecan, tegafur-gimeracil-oteracil potassium, sapacitabine); tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib, gefitinib, cetuximab, panitumumab, necitumumab, PF-00299804, nimotuzumab, RO5083945), MET inhibitor (eg, PF-02341066, ARQ 197), PI3K kinase inhibitor (eg, XL147, GDC-0941), Raf/MEK dual kinase inhibitor (eg, RO5126766), PI3K/mT0R dual kinase inhibitor (eg, XL765), SRC inhibitor (eg, dasatinib), dual inhibitor (eg, BIBW 2992, GSK1363089, ZD6474, AZD0530 , AG-013736, lapatinib, MEHD7945A, linifanib), multikinase inhibitor (eg, sorafenib, sunitinib, pazopanib, AMG 706, XL184, MGCD265, BMS-690514, R935788), VEGF inhibitor (eg, endostar, endostatin, bevacizumab, cediranib, BIBF 1120, axitinib, tivozanib, AZD2171), cancer vaccine (eg, BLP25 liposomal vaccine, GVAX, recombinant DNA and adenovirus expressing L523S protein), Bcl-2 inhibitor (eg, oblimersen sodium), proteasome inhibitor (eg, bortezomib, carfilzomib , NPI-0052, MLN97 08), paclitaxel or paclitaxel-based agent, docetaxel, IGF-1 receptor inhibitor (eg, cixutumumab, MK-0646, OSI 906, CP-751,871, BIIB022), hydroxychloroquine, HSP90 inhibitor (eg, tanespymycin, STA-9090, AUY922 , XL888), mTOR inhibitor (eg, everolimus, temsirolimus, ridaforolimus), Ep-CAM/CD3 bispecific antibody (eg, MT110), CK-2 inhibitor (eg, CX-4945), HDAC inhibitor (eg, MS 275, LBH589, vorinostat, valproic acid, FR901228), DHFR inhibitor (eg, pralatrexate), retinoid (eg, bexarotene, tretinoin), antibody drug conjugate (eg, SGN-15), bisphosphonate (eg, zoledronic acid), cancer vaccine (eg, belagenpumatucel-L), low molecular weight heparin (LMWH) (eg, tinzaparin, enoxaparin), GSK1572932A, melatonin, talactoferrin, dimesna, topoisomerase inhibitor (eg, amrubicin, etoposide, carenitecin), nelfinavir, cilengitide, ErbB3 inhibitor (eg, MM-121, U3-1287), inhibition survivin tor (eg, YM155, LY2181308), eribulin mesylate, COX-2 inhibitor (eg, celecoxib), pegfilgrastim, Polo-like kinase 1 inhibitor (eg, BI 6727), TRAIL receptor 2 (TR-2) agonist (eg , CS-1008), CNGRC Peptide Conjugate (SEQ ID NO:225) TNF-alpha, dichloroacetate (DCA), HGF inhibitor (e.g. SCH 900105), SAR240550, PPAR-gamma agonist (e.g. secretase (eg, RO4929097), epigenetic therapy (eg, 5-azacytidine), nitroglycerin, MEK inhibitor (eg, AZD6244), cyclin-dependent kinase inhibitor (eg, UCN-01), cholesterol-Fus1, antimicrotubulin agent (eg, E7389 ), farnesyl OH transferase inhibitor (eg, lonafarnib), immunotoxin (eg, BB-10901, SS1 (dsFv) PE38), fondaparinux, vasoconstrictor (eg, AVE8062), PD-L1 inhibitor (eg, MDX- 1105, MDX-1106), beta-glucan, NGR-hTNF, EMD 521873, MEK inhibitor (eg GSK1120212), epothilone analogue (eg ixabepilone), inhibitor β kinesin spindle (eg, 4SC-205), telomere targeting agent (eg, KML-001), P70 pathway inhibitor (eg, LY2584702), AKT inhibitor (eg, MK-2206), angiogenesis inhibitor (eg, lenalidomide ), a Notch signaling pathway inhibitor (eg, OMP-21M18), radiation therapy, surgery, and combinations thereof.

An example of suitable therapeutic agents for use in combination with anti-PD-1 antibody molecules, alone or in combination with another immunomodulator (e.g. anti-LAG-3, anti-PD-L1 or anti-TIM-3 antibody molecule), for the treatment of ovarian cancer includes, without limitation, a chemotherapeutic agent (e.g., paclitaxel or paclitaxel-based agent; docetaxel; carboplatin; gemcitabine; doxorubicin; topotecan; cisplatin, irinotecan, TLK286, ifosfamide, olaparib, oxaliplatin, melphalan, disodium pemetrexed, SJG-136, cyclophosphamide, etoposide , decitabine); ghrelin antagonist (eg, AEZS-130), immunotherapy (eg, APC8024, oregovomab, ORT-821), tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib), dual inhibitor (eg, E7080), multikinase inhibitor (eg, AZD0530 , JI-101, sorafenib, sunitinib, pazopanib), ON 01910.Na), VEGF inhibitor (eg, bevacizumab, BIBF 1120, cediranib, AZD2171), PDGFR inhibitor (eg, IMC-3G3), paclitaxel, topoisomerase inhibitor (eg, carenitecin, irinotecan), HDAC inhibitor (eg, valproate, vorinostat), folate receptor inhibitor (eg, farletuzumab), angiopoietin inhibitor (eg, AMG 386), epothilone analog (eg, ixabepilone), proteasome inhibitor (eg, carflizomib), inhibitor IGF-1 receptor (eg, OSI 906, AMG 479), PARP inhibitor (eg, veliparib, AG014699, iniparib, MK-4827), Aurora kinase inhibitor (eg, MLN8237, ENMD-2076), angiogenesis inhibitor (eg, lenalidomide ), DHFR inhibitor (e.g. pralatrexate), radioimmuno notherapy (eg, Hu3S193), statin (eg, lovastatin), topoisomerase 1 inhibitor (eg, NKTR-102), cancer vaccine (eg, p53 long peptide vaccine, autologous OC-DC vaccine), mTOR inhibitor (eg, temsirolimus, everolimus), BCR/ABL inhibitor (eg, imatinib), ET-A receptor antagonist (eg, ZD4054), TRAIL receptor 2 (TR-2) agonist (eg, CS-1008), HGF/SF inhibitor (eg. , AMG 102), EGEN-001, Polo-like kinase 1 inhibitor (eg BI 6121), gamma secretase inhibitor (eg RO4929097), Wee-1 inhibitor (eg MK-1775), antimicrotubulin agent (eg vinorelbine , E7389), immunotoxin (eg, denileukin diftitox), SB-485232, vascular disruptor (eg, AVE8062), integrin inhibitor (eg, EMD 525797), kinesin spindle inhibitor (eg, 4SC-205), revlimid, HER2 inhibitor (eg MGAH22), ErrB3 inhibitor (eg MM-121), radiotherapy; and their combinations.

In one embodiment, an anti-PD-1 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-L1, or anti-TIM-3 antibody molecule), is used to treat myeloma, alone or in combination with one or more of: chemotherapy or other anticancer agents (e.g. thalidomide analogues e.g. lenalidomide), HSCT (Cook, R. (2008) J Manag Care Pharm. 14(7 Suppl):19-25), anti-TIM-3 antibodies (Hallett, WHD et al. (2011) J of American Society for Blood and Marrow Transplantation 17(8):1133-145), tumor antigen-sensitized dendritic cells, fusions (eg, electrofusions) of tumor cells and dendritic cells, or idiotype vaccinations immunoglobulin produced by malignant plasma cells (reviewed by Yi, Q. (2009) Cancer J. 15(6):502-10).

In yet another embodiment, an anti-PD-1 antibody molecule, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-L1, or anti-TIM-3 antibody molecule), is used to treat kidney cancer, e.g. cell carcinoma (RCC), non-clear cell renal cell carcinoma, or metastatic RCC. An anti-PD-1 antibody molecule can be used in combination with one or more of: an immune strategy (eg, interleukin-2 or interferon-α), a targeting agent (eg, a VEGF inhibitor such as an anti-VEGF monoclonal antibody, such as bevacizumab ( Rini, B. I. et al (2010) J Clin Oncol 28(13):2137-2143)); a VEGF tyrosine kinase inhibitor such as sunitinib, sorafenib, axitinib, and pazopanib (reviewed by Pal. S.K. et al. (2014) Clin. Advances in Hematology & Oncology 12(2):90-99)); an RNAi inhibitor) or an inhibitor of a downstream VEGF signaling mediator such as the mammalian target of rapamycin (mTOR) inhibitor such as everolimus and temsirolimus (Hudes, G. et al. (2007) N. Engl. J. Med. 356(22): 2271-2281, Motzer, R. J. et al (2008) Lancet 372: 449-456).

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of chronic myelogenous leukemia (AML) according to the invention includes, without limitation, a chemotherapeutic agent (such as cytarabine, hydroxyurea, clofarabine, melphalan, thiotepa, fludarabine, busulfan, etoposide, cordycepin, pentostatin, capecitabine , azacitidine, cyclophosphamide, cladribine, topotecan), tyrosine kinase inhibitor (eg, BCR/ABL inhibitor (eg, imatinib, nilotinib), ON 01910.Na, dual inhibitor (eg, dasatinib, bosutinib), multikinase inhibitor (eg, DCC-2036 , ponatinib, sorafenib, sunitinib, RGB-286638)), interferon-alpha, steroids, apoptotic agent (eg, omacetaxin mepesuccinate), immunotherapy (eg, allogeneic CD4+ Th1-like memory T cells/microparticle-bound anti-CD3/anti-CD28 antibodies, autologous cytokine-induced killer (CIK), AHN-12), CD52 targeting agent (eg, alemtuzumab) , HSP90 inhibitor (eg tanespimycin, STA-9090, AUY922, XL888), mTOR inhibitor (eg everolimus), SMO antagonist (eg BMS 833923), ribonucleotide reductase inhibitor (eg 3-AP), JAK-2 inhibitor (eg , INCB018424), hydroxychloroquine, retinoid (eg, fenretinide), cyclin-dependent kinase inhibitor (eg, UCN-01), HDAC inhibitor (eg, belinostat, vorinostat, JNJ-26481585), PARP inhibitor (eg, veliparib), MDM2 antagonist (eg, RO5045337), Aurora-B kinase inhibitor (eg, TAK-901), radioimmunotherapy (eg, actinium-225 labeled anti-CD33 antibody, HuM195), Hedgehog inhibitor (eg, PF-04449913), STAT3 inhibitor (eg, ODS -31121), KB004, cancer vaccine (e.g. AG858 ), bone marrow transplantation, stem cell transplantation, radiation therapy, and combinations thereof.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of chronic lymphocytic leukemia (CLL) includes, but is not limited to, a chemotherapeutic agent (eg, fludarabine, cyclophosphamide, doxorubicin, vincristine, chlorambucil, bendamustine, chlorambucil, busulfan, gemcitabine, melphalan, pentostatin, mitoxantrone , 5-azacytidine, pemetrexed disodium), tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, erlotinib), BTK inhibitor (eg, PCI-32765), multikinase inhibitor (eg, MGCD265, RGB-286638), CD-targeting agent) -20 (eg, rituximab, ofatumumab, RO5072759, LFB-R603), CD52 targeting agent (eg, alemtuzumab), prednisolone , darbepoetin alfa, lenalidomide, Bcl-2 inhibitor (eg, ABT-263), immunotherapy (eg, allogeneic CD4+ Th1-like memory T cells/microparticle-associated anti-CD3/anti-CD28 antibodies, autologous cytokine-induced killer cells ( CIK)), HDAC inhibitor (eg, vorinostat, valproic acid, LBH589, JNJ-26481585, AR-42), XIAP inhibitor (eg, AEG35156), CD-74 targeting agent (eg, milatuzumab), mTOR inhibitor ( e.g. everolimus), AT-101, immunotoxin (e.g. CAT-8015, anti-Tac(Fv)-PE38 antibody (LMB-2)), CD37 targeting agent (e.g. TRU-016), radioimmunotherapy (e.g. I131-tositumomab), hydroxychloroquine, perifosine, SRC inhibitor (eg, dasatinib), thalidomide, PI3K delta inhibitor (eg, CAL-101), retinoid (eg, fenretinide), MDM2 antagonist (eg, RO5045337), plerixaphor, Aurora- kinases (eg, MLN8237, TAK-901), proteasome inhibitor (eg, bortezomib), vo, CD-19 targeting (eg, MEDI-551, MOR208), MEK inhibitor (eg, ABT-348), JAK-2 inhibitor (eg, INCB018424), hypoxia-activated prodrug (eg, TH-302), paclitaxel or a paclitaxel-based agent, an HSP90 inhibitor, an AKT inhibitor (eg, MK2206), an HMG-CoA reductase inhibitor (eg, simvastatin), GNKG186, radiation therapy, bone marrow transplantation, stem cell transplantation, and a combination thereof.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of acute lymphocytic leukemia (ALL), includes, but is not limited to, a chemotherapeutic agent (eg, prednisolone, dexamethasone, vincristine, asparaginase, daunorubicin, cyclophosphamide, cytarabine, etoposide, thioguanine, mercaptopurine, clofarabine, liposomal annamycin, busulfan, etoposide, capecitabine, decitabine, azacitidine, topotecan, temozolomide), tyrosine kinase inhibitor (eg, BCR/ABL inhibitor (eg, imatinib, nilotinib), ON 01910.Na, multikinase inhibitor (eg, sorafenib)), CD-20 targeting agent (eg, rituximab), CD52 targeting agent (eg, alemtuzumab), inhibitor HSP90 (eg, STA-9090), mTOR inhibitor (eg, everolimus, rapamycin), JAK-2 inhibitor (eg, INCB018424), HER2/Neu receptor inhibitor (eg, trastuzumab), proteasome inhibitor (eg, bortezomib), methotrexate, asparaginase, CD-22 targeting agent (eg, epratuzumab, inotuzomab), immunotherapy (autologous cytokine-induced killer (CIK), AHN-12), blinatumomab, cyclin-dependent kinase inhibitor (eg, UCN-01) , CD45 targeting agent (eg, BC8), MDM2 antagonist (eg, RO5045337), immunotoxin (eg, CAT-8015, DT2219ARL), HDAC inhibitor (eg, JNJ-26481585), JVRS-100, paclitaxel, or based on paclitaxel, a STAT3 inhibitor (eg OPB-31121), a PARP inhibitor (eg veliparib), EZN-2285, radiation therapy, a steroid, bone marrow transplantation, stem cell transplantation, or a combination thereof.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of acute myeloid leukemia (AML), includes, but is not limited to, a chemotherapeutic agent (e.g., cytarabine, daunorubicin, idarubicin, clofarabine, decitabine, vozaroxin, azacitidine, clofarabine, ribavirin, CPX-351, treosulfan, elacytarabine, azacitidine), tyrosine kinase inhibitor (eg, BCR/ABL inhibitor (eg, imatinib, nilotinib), ON 01910.Na, multikinase inhibitor (eg, midostaurin, SU 11248, quisartinib, sorafinib)), immunotoxin (eg, gemtuzumab ozogamicin), DT388IL3 fusion protein, HDAC inhibitor (eg, vorinostat, LBH589), plerixaphor, mTOR inhibitor (eg, everolimus), SRC inhibitor (eg, dasatinib), inhibition HSP90 inhibitor (eg, STA-9090), retinoid (eg, bexarotene, Aurora kinase inhibitor (eg, BI 811283), JAK-2 inhibitor (eg, INCB018424), Polo-like kinase inhibitor (eg, BI 6727), cenersen , CD45 targeting agent (eg, BC8), cyclin-dependent kinase inhibitor (eg, UCN-01), MDM2 antagonist (eg, RO5045337), mTOR inhibitor (eg, everolimus), LY573636-sodium, ZRx-101, MLN4924, lenalidomide, immunotherapy (eg, AHN-12), histamine dihydrochloride, radiation therapy, bone marrow transplantation, stem cell transplantation, and a combination thereof.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of multiple myeloma (MM) includes, but is not limited to, a chemotherapeutic agent (e.g., melphalan, amifostine, cyclophosphamide, doxorubicin, clofarabine, bendamustine, fludarabine, adriamycin, SyB L-0501), thalidomide, lenalidomide, dexamethasone, prednisone, pomalidomide, proteasome inhibitor (eg, bortezomib, carfilzomib, MLN9708), cancer vaccine (eg, GVAX), CD-40 targeting agent (eg, SGN-40, CHIR-12.12), perifosine, zoledronic acid, immunotherapy (eg, MAGE-A3, NY-ESO-1, HuMax-CD38), HDAC inhibitor (eg, vorinostat, LBH589, AR-42), aplidine, cyclin-dependent kinase inhibitor (eg. , PD-0332991, dinaciclib), arsenic trioxide, CB3304, HSP90 inhibitor (eg, KW-2478), tyrosine kinase inhibitor (eg, EGFR inhibitor (eg, cetuximab), multikinase inhibitor (eg, AT9283)), VEGF inhibitor (eg, bevacizumab), plerixaphor, MEK inhibitor (eg, AZD6244), IPH2101, atorvastatin, immunotoxin (eg, BB-10901), NPI-0052, radioimmunotherapy (eg, yttrium Y90 ibritumomab tiuxetan), STAT3 inhibitor (eg, OPB-31121) , MLN4924, an Aurora kinase inhibitor (eg, ENMD-2076), IMGN901, ACE-041, a CK-2 inhibitor (eg, CX-4945), radiation therapy, bone marrow transplantation, stem cell transplantation, and a combination thereof.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of prostate cancer includes, but is not limited to, a chemotherapeutic agent (eg, docetaxel, carboplatin, fludarabine), abiraterone, hormone therapy (eg, flutamide, bicalutamide, nilutamide, cyproterone acetate, ketoconazole, aminoglutethimide , abarelix, degarelix, leuprolide, goserelin, triptorelin, buserelin), tyrosine kinase inhibitor (eg, dual kinase inhibitor (eg, lapatanib), multikinase inhibitor (eg, sorafenib, sunitinib)), VEGF inhibitor (eg, bevacizumab), TAK-700 , cancer vaccine (eg, BPX-101, PEP223), lenalidomide, TOK-001, IGF-1 receptor inhibitor (eg, cixutumumab), TR C105, Aurora kinase inhibitor (eg, MLN8237), proteasome inhibitor (eg, bortezomib), OGX-011, radioimmunotherapy (eg, HuJ591-GS), HDAC inhibitor (eg, valproic acid, SB939, LBH589), hydroxychloroquine, mTOR inhibitor (eg, everolimus), dovitinib lactate, diindolylmethane, efavirenz, OGX-427, genistein, IMC-3G3, bafetinib, CP-675,206, radiation therapy, surgery, or a combination thereof.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of HNSCC includes, but is not limited to, one or both of Compound A8 described herein (or compound described in PCT publication WO2010/029082) and cetuximab (e.g., Erbitux available from BMS) . In some embodiments, the therapeutic agent (eg, Compound A8 or an A8-related compound) is a PI3K modulator, eg, a PI3K inhibitor. In some embodiments, the therapeutic agent (eg, cetuximab) modulates, eg, inhibits, EGFR. In some embodiments, the cancer has, or is identified as having, elevated levels or activity of PI3K or EGFR compared to a control cell or reference value.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of gastric cancer, for example, MSI-H and/or EBV+ gastric cancer, includes, without limitation, Compound A8 described in this application (or the compound described in PCT publication WO2010/029082) . In some embodiments, the therapeutic agent (eg, Compound A8 or an A8-related compound) is a PI3K modulator, eg, a PI3K inhibitor. In some embodiments, the cancer has, or is identified as having, elevated PI3K levels or activity compared to a control cell or reference value.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of gastric cancer, for example, MSI-H and/or RNF43-inactivated gastric cancer, includes, without limitation, Compound A28 described in this application (or the compound described in PCT publication WO2010/ 101849). In some embodiments, the therapeutic agent (eg Compound A28 or an A28 related compound) is a modulator, eg an inhibitor, of porcupine. In some embodiments, the cancer has, or is identified as having, increased levels or activity of porcupin compared to a control cell or reference value.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of gastrointestinal stromal tumor (GIST), includes, but is not limited to, Compound A16 described in this application (or the compound described in PCT publication WO1999/003854). In some embodiments, the therapeutic agent (eg Compound A16 or an A16 related compound) is a modulator, eg an inhibitor, of tyrosine kinase. In some embodiments, the cancer has, or is determined to have, elevated tyrosine kinase levels or activity compared to a control cell or reference value.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of NSCLC, for example, squamous or adenocarcinoma, includes, but is not limited to, one or both of Compound A17 described in this application (or the compound described in US patent 7,767,675 and 8,420,645) and Compound A23 described in this application (or the compound described in PCT publication WO2003/077914). In some embodiments, a compound (eg Compound A17 or an A17 related compound) modulates, eg inhibits, c-MET. In some embodiments, a compound (eg, Compound A23 or an A23-related compound) modulates, eg, inhibits, Alk. In some embodiments, the cancer has, or is determined to have, elevated levels or activity of one or both of c-MET or Alk compared to a control cell or reference value. In some embodiments, the cancer has, or is identified as having, a mutation in EGFR.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of melanoma (eg, NRAS melanoma), includes, but is not limited to, one or both of Compound A24 described herein (or the compound described in US Pat. Nos. 8,415,355 and 8,685,980) and Compound A34 described in this application (or the compound described in PCT publication WO2003/077914). In some embodiments, a compound (eg, Compound A24 or a compound associated with A24) modulates, eg, inhibits, one or more of JAK and CDK4/6. In some embodiments, a compound (eg, Compound A34 or an A34-related compound) modulates, eg, inhibits, MEK. In some embodiments, the cancer has, or is identified as having, elevated levels or activity of one or more of JAK, CDK4/6, and MEK compared to a control cell or reference value.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3), for the treatment of melanoma (eg, NRAS melanoma), includes, but is not limited to, one or both of Compound A29 described herein (or compound described in PCT publication WO2011/025927) and Compound A34 described in this application (or the compound described in PCT publication WO2003/077914). In some embodiments, a compound (eg, Compound A29 or an A29-related compound) modulates, eg, inhibits, BRAF. In some embodiments, a compound (eg, Compound A34 or an A34-related compound) modulates, eg, inhibits, MEK. In some embodiments, the cancer has, or is identified as having, elevated levels or activity of one or both of BRAF and MEK compared to a control cell or reference value.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3) for the treatment of squamous NSCLC includes, but is not limited to, Compound A5 described herein (or the compound described in US Pat. No. 8,552,002). In some embodiments, a compound (eg, Compound A5 or an A5-related compound) modulates, eg, inhibits, FGFR. In some embodiments, the cancer has, or is identified as having, elevated FGFR levels or activity compared to a control cell or reference value.

An example of suitable therapeutic agents for use in the combinations described herein, e.g., combinations with anti-PD-1 antibody molecules described herein, alone or in combination with another immunomodulator (e.g., anti-LAG-3, anti-PD- L1 or against TIM-3) for the treatment of colon and rectal cancer includes, but is not limited to, one or both of Compound A29 described herein (or a compound in PCT publication WO2011/025927) and cetuximab (e.g., Erbitux available from BMS ). In some embodiments, the therapeutic agent (eg Compound A29 or an A29 related compound) modulates, eg inhibits, BRAF. In some embodiments, the therapeutic agent (eg, cetuximab) modulates, eg, inhibits, EGFR. In some embodiments, the cancer has, or is identified as having, elevated levels or activity of BRAF or EGFR compared to a control cell or reference value.

Also provided herein is a method of treating cancer with Compound A8, cetuximab, and a combination as described herein, such as combinations with the anti-PD-1 antibody molecules described herein (optionally in combination with an anti-TIM-3 antibody molecule or an anti-PD-1 antibody molecule to LAG-3). In some embodiments, the patient is first treated with Compound A8 and cetuximab. This treatment is continued for a period of time, such as a set period of time, such as about 1, 2, 4, 6, 8, 10 or 12 months. An anti-PD-1 antibody molecule is then administered (optionally in combination with an anti-TIM-3 antibody molecule or an anti-LAG-3 antibody molecule). An anti-PD-1 antibody may optionally be used in combination with cetuximab.

In some embodiments, the patient is first treated with all three of Compound A8, cetuximab, and an anti-PD-1 antibody molecule (optionally in combination with an anti-TIM-3 antibody or anti-LAG-3 antibody molecule). This treatment is continued for a period of time, such as a set period of time, such as about 6, 8, 10 or 12 months. The administration of Compound A8 and/or cetuximab can then be tapered such that the maintenance phase includes treatment with a PD-1 antibody molecule (eg, alone or in combination with an anti-TIM-3 antibody or an anti-LAG-3 antibody molecule), but not includes treatment with Compound A8 or cetuximab.

In other embodiments, three compounds (Compound A8, cetuximab, and an anti-PD-1 antibody molecule, optionally in combination with an anti-TIM-3 antibody or an anti-LAG-3 antibody molecule) are used sequentially at the start of treatment. For example, Compound A8 and cetuximab may be used initially as described above. An anti-PD-1 antibody molecule (optionally in combination with an anti-TIM-3 antibody or an anti-LAG-3 antibody molecule) is then included in the application schedule. Then the administration of Compound A8 and/or cetuximab can be reduced as described above.

Approximate dosages for three (or more) agent regimens are as follows. The anti-PD-1 antibody molecule may be administered, for example, at a dose of about 1-40 mg/kg, for example, 1-30 mg/kg, for example, about 5-25 mg/kg, about 10-20 mg/kg, about 1 -5 mg/kg or approximately 3 mg/kg. Alternatively, the anti-PD-1 antibody molecule may be administered, for example, at a dose of about 200-500 mg, such as about 300-500 mg, about 200-400 mg, about 250-350 mg, or about 300-400 mg, or about 300 mg, or approximately 400 mg. In one embodiment, the anti-PD-1 antibody molecule is administered once every three weeks, for example at a dose of 300 mg once every three weeks. In another embodiment, the anti-PD-1 antibody molecule is administered once every four weeks, for example at a dose of 400 mg once every four weeks. In some embodiments, Compound A8 is administered at a dose of about 200-300, 300-400, or 200-400 mg. In some embodiments, cetuximab is administered at an initial dose of 400 mg/m ² as a 120-minute intravenous infusion, followed by a 250 mg/m ² weekly infusion over 60 minutes. In embodiments, one or more of Compound A8, cetuximab, and a PD-1 antibody molecule is administered at a dose that is lower than the dose at which such an agent would normally be used in monotherapy, e.g., by about 0-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80% or 80-90% lower than the dose at which such an agent is usually used in monotherapy. In embodiments, one or more of Compound A8, cetuximab, and a PD-1 antibody molecule is administered at a dose that is lower than the dose of such an agent specified in this paragraph, for example, by about 0-10%, 10-20%, 20- 30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90% lower than the dose of such agent referred to in this paragraph. In some embodiments, the concentration of Compound A8 that is required for inhibition, such as growth inhibition, is lower when Compound A8 is used in combination with one or both of cetuximab and a PD-1 antibody molecule than when Compound A8 is used alone. In some embodiments, the concentration of cetuximab required for inhibition, such as growth inhibition, is lower when cetuximab is used in combination with one or both of Compound A8 and a PD-1 antibody molecule than when cetuximab is used alone. In some embodiments, the concentration of the PD-1 antibody molecule that is required for inhibition, e.g., growth inhibition, is lower when the PD-1 antibody molecule is used in combination with one or both of cetuximab and Compound A8 than when the PD-1 antibody molecule is 1 is applied separately.

The present application further discloses a method of treating cancer with anti-PD-1 antibody molecules, alone or in combination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-L1, or anti-TIM-3 antibody molecule) and a targeted antitumor agent, for example, an agent that targets one or more proteins. In some embodiments, the anti-PD-1 antibody molecule (and optionally other immunomodulator(s)) is administered first and the targeted anticancer agent is administered second. The time period between administration of the anti-PD-1 antibody molecule and the targeted antitumor agent may be, for example, 10, 20, or 30 minutes, 1, 2, 4, 6, or 12 hours, or 1, 2, 3, 4, 5, 6, or 7 days, or any period of time within the specified range. In some embodiments, the anti-PD-1 antibody molecule is administered multiple times over a period of time (e.g., 1, 2, 3, 4, 5, or 6 days, or 1, 2, 4, 8, 12, 16, or 20 weeks, or any a period of time in the specified range) before the introduction of an antitumor agent of directed action. In other embodiments, the anti-PD-1 antibody molecule and the targeted anticancer agent are administered at substantially the same time.

Infectious diseases

Other methods of the invention are used to treat patients who have been exposed to certain toxins or pathogens. Thus, in another aspect of the present invention, there is provided a method of treating an infectious disease in a subject, comprising administering to the subject a combination disclosed herein, for example, a combination comprising an anti-PD-1 antibody molecule, thereby treating the infectious disease in the subject.

In the treatment of infection (eg, acute and/or chronic), the administration of anti-PD-1 antibody molecules can be combined with standard therapies in addition to or instead of stimulating the host's natural immune defenses against infection. Natural host defense mechanisms against infection include, but are not limited to, inflammation, fever, antibody-mediated host defense, T-lymphocyte-mediated host defense mechanisms, including lymphokine secretion, and cytotoxic T cells (particularly during viral infection). ), complement-mediated lysis and opsonization (facilitating phagocytosis) and phagocytosis. The ability of anti-PD-1 antibody molecules to reactivate dysfunctional T cells may be useful in the treatment of chronic infections, in particular those in which cellular immunity is essential for full recovery.

Similar to use in tumors as discussed above, antibody-mediated blockade of PD-1 can be used alone or as an adjuvant, in combination with vaccines, to stimulate an immune response to pathogens, toxins, and self-antigens. Examples of pathogens for which such a therapeutic approach may be particularly useful include pathogens for which no effective vaccine currently exists or pathogens for which conventional vaccines are not fully effective. Such pathogens include, but are not limited to, HIV, hepatitis (A, B, and C), influenza, herpes, giardia, Plasmodium malaria, Leishmania, Staphylococcus aureus , Pseudomonas Aeruginosa . Blockade of PD-1 is particularly useful against infections caused by pathogens such as HIV that present altered antigens during infections. These new epitopes are recognized as foreign at the time of administration of the anti-human PD-1 antibody, which elicits a strong T cell response that is not suppressed by negative signals through PD-1.

Complementary and other combination therapies

Molecules of antibodies against PD-1 can be used in any of the combinations disclosed in this application. In some embodiments, this application provides combinations with one or more additional therapies. Many of the combinations in this section can be used in the treatment of cancer, but other indications are also described. In some embodiments, the anti-PD-1 antibody molecule is administered by injection (e.g., subcutaneously or intravenously) at a dose (e.g., constant dose) of about 200 mg to 500 mg, for example, about 300 mg to 400 mg or about 300 mg, or approximately 400 mg. The administration schedule (eg, fixed dose schedule) may vary, for example, from once a week to once every 2, 3, 4, 5 or 6 weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 300 mg once every three weeks. In one embodiment, the anti-PD-1 antibody molecule is administered at a dose of about 400 mg once every four weeks. This section focuses on combinations of anti-PD-1 antibody molecules, optionally in combination with one or more immunomodulators (e.g., an anti-TIM-3 antibody molecule, an anti-LAG-3 antibody molecule, or an anti-PD-L1 antibody molecule), with one or more more than those described in Table 7. In the combinations below, in one embodiment, the anti-PD-1 antibody molecule comprises:

(a) a heavy chain variable region (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 light chain variable region (VL) comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33;

(b) VH comprising the amino acid sequence of VHCDR1 selected from SEQ ID NO: 1; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32;

(c) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224, the amino acid sequence of VHCDR2 SEQ ID NO: 5, and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33; or

(d) VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32.

In the combinations below, in another embodiment, the anti-PD-1 antibody molecule comprises: (i) a heavy chain variable region (VH) comprising the VHCDR1 amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 4, or SEQ ID NO: : 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 or SEQ ID NO: 5; and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and (ii) a light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 10 or SEQ ID NO: 13, the VLCDR2 amino acid sequence of SEQ ID NO: 11 or SEQ ID NO: 14, and the VLCDR3 amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 33.

In one embodiment, the combination, for example, a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a PKC inhibitor, sotrastaurine (Compound A1), or a compound disclosed in PCT Publication WO 2005/039549 , for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the PKC inhibitor is sotrastaurine (Compound A1) or a compound disclosed in PCT publication WO 2005/039549. In one embodiment, a PD-1 antibody molecule is used in combination with sotrastaurine (Compound A1) or a compound described in PCT publication WO 2005/039549 to treat a disorder such as cancer, melanoma, non-Hodgkin's lymphoma, inflammatory bowel disease, transplant rejection, eye disorder or psoriasis.

In some embodiments, sotrastaurine (Compound A1) is administered at a dose of about 20-600 mg, e.g., about 200 to about 600 mg, about 50 mg to about 450 mg, about 100 mg to 400 mg, about 150 mg to 350 mg, or about 200 mg to about 300 mg, such as about 50 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg, 500 mg, or 600 mg. The administration schedule may vary, for example, from once every two days to once a day, two or three times a day.

In one embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a BCR-ABL inhibitor, Tasigna® (Compound A2), or a compound disclosed in PCT publication WO 2004/005281, for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the BCR-ABL inhibitor is Tasigna or a compound disclosed in PCT publication WO 2004/005281. In one embodiment, a PD-1 antibody molecule is used in combination with Tasigna (Compound A2) or a compound described in PCT publication WO 2004/005281 to treat a disorder such as lymphocytic leukemia, Parkinson's disease, neurological cancer, melanoma, cancer of the digestive/ gastrointestinal tract, colon and rectal cancer, myeloid leukemia, head and neck cancer, or pulmonary hypertension.

In one embodiment, the BCR-ABL inhibitor or Tasigna is administered at a dose of about 300 mg (e.g., twice daily, e.g., in the case of newly diagnosed Ph+ CML-CP) or about 400 mg, e.g., twice daily, e.g. case of resistant or intolerant Ph+ CML-CP and CML-AP). The BCR-ABL inhibitor or Compound A2 is administered at a dose of approximately 300-400 mg.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an HSP90 inhibitor such as 5-(2,4-dihydroxy-5-isopropylphenyl)-N -ethyl-4-(4-(morpholinomethyl)phenyl)isoxazole-3-carboxamide (Compound A3), or a compound disclosed in PCT Publication WO 2010/060937 or WO 2004/072051, for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the HSP90 inhibitor is 5-(2,4-dihydroxy-5-isopropylphenyl)-N-ethyl-4-(4-(morpholinomethyl)phenyl)-isoxazole-3-carboxamide (Compound A3) or a compound disclosed in PCT publications WO 2010/060937 or WO 2004/072051. In one embodiment, the PD-1 antibody molecule is used in combination with 5-(2,4-dihydroxy-5-isopropylphenyl)-N-ethyl-4-(4-(morpholinomethyl)phenyl)isoxazole-3-carboxamide (Compound A3) or a compound described in PCT Publication WO 2010/060937 or WO 2004/072051 for the treatment of a disorder such as cancer, multiple myeloma, non-small cell lung cancer, lymphoma, gastric cancer, breast cancer, digestive/gastrointestinal cancer, cancer pancreas, colon and rectal cancer, solid tumor or hematopoietic disorder.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, is used in combination with a PI3K and/or mTOR inhibitor, dactolisib (Compound A4), or 8-(6-methoxy-pyridine -3-yl)-3-methyl-1-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-1,3-dihydro-imidazo[4,5-c]quinolin-2-one (Compound A41 ), or a compound disclosed in PCT Publication WO 2006/122806, for the treatment of a disorder, for example the disorder described herein. In one embodiment, the PI3K and/or mTOR inhibitor is dactolisib (Compound A4), 8-(6-methoxy-pyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3-trifluoromethyl -phenyl)-1,3-dihydro-imidazo[4,5-c]quinolin-2-one (Compound A41), or the compound disclosed in PCT Publication WO 2006/122806. In one embodiment, the PD-1 antibody molecule is used in combination with dactolisib (Compound A4), 8-(6-methoxy-pyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3- trifluoromethyl-phenyl)-1,3-dihydro-imidazo[4,5-c]quinolin-2-one (Compound A41), or a compound described in PCT Publication WO 2006/122806, for the treatment of a disorder such as cancer, cancer prostate, leukemia (eg, lymphocytic leukemia), breast cancer, brain cancer, bladder cancer, pancreatic cancer, kidney cancer, solid tumor, or liver cancer.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an FGFR inhibitor, 3-(2,6-dichloro-3,5-dimethoxyphenyl)-1 -(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)-pyrimidin-4-yl)-1-methylurea (Compound A5), or a compound disclosed in US Pat. No. 8,552,002, for the treatment of a disorder, for example, the violation described in this application. In one embodiment, the FGFR inhibitor is 3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl )-1-methylurea (Compound A5) or the compound disclosed in US Patent 8,552,002. In one embodiment, a PD-1 antibody molecule is used in combination with Compound A5 or a compound described in US 8,552,002 to treat a disorder such as digestive/gastrointestinal cancer, hemoblastosis, or a solid tumor.

In one embodiment, an FGFR inhibitor or 3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl )-1-methylurea (Compound A5) is administered at a dose of about 100-125 mg (eg per day), eg about 100 mg or about 125 mg.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a PI3K inhibitor, buparlisib (Compound A6), or a compound disclosed in PCT Publication WO 2007/084786 , for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the PI3K inhibitor is buparlisib (Compound A6) or a compound disclosed in PCT publication WO 2007/084786. In one embodiment, a PD-1 antibody molecule is used in combination with buparlisib (Compound A6) or a compound disclosed in PCT Publication WO 2007/084786 to treat a disorder such as prostate cancer, non-small cell lung cancer, endocrine cancer, leukemia, cancer ovarian cancer, melanoma, bladder cancer, breast cancer, cancer of the female reproductive system, digestive/gastrointestinal cancer, colon and rectal cancer, glioblastoma multiforme, solid tumor, non-Hodgkin's lymphoma, hematopoietic disorder, or head and neck cancer.

In one embodiment, the PI3K inhibitor or buparlisib (Compound A6) is administered at a dose of about 100 mg (eg, per day).

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an FGFR inhibitor, 8-(2,6-difluoro-3,5-dimethoxyphenyl)-N -(4-(dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin-5-carboxamide (Compound A7), or a compound disclosed in PCT publication WO 2009/141386, for the treatment of a disorder, e.g., the disorder described in this application. In one embodiment, the FGFR inhibitor is 8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-(dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin-5-carboxamide (Compound A7 ) or a compound disclosed in PCT publication WO 2009/141386. In one embodiment, the FGFR inhibitor is 8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-(dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin-5-carboxamide (Compound A7 ). In one embodiment, the PD-1 antibody molecule is used in combination with 8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-(dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin- 5-carboxamide (Compound A7) or a compound disclosed in PCT publication WO 2009/141386 for the treatment of a disorder such as cancer characterized by angiogenesis.

In one embodiment, an FGFR inhibitor or 8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-(dimethylamino)methyl)-1H-imidazol-2-yl)quinoxalin-5-carboxamide (Compound A7 ) is administered at a dose of, for example, from about 3 mg to about 5 g, more preferably from about 10 mg to about 1.5 g per person per day, optionally divided into 1-3 separate doses, which may, for example, have the same size.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a PI3K inhibitor, (S)-N1-(4-methyl-5-(2-( 1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2-yl)pyrrolidin-1,2-dicarboxamide (Compound A8), or a compound disclosed in PCT publication WO 2010/ 029082, for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the PI3K inhibitor is (S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2- yl) pyrrolidine-1,2-dicarboxamide (Compound A8) or the compound disclosed in PCT Publication WO 2010/029082. In one embodiment, the PD-1 antibody molecule is used in combination with (S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4- yl)thiazol-2-yl)pyrrolidine-1,2-dicarboxamide (Compound A8) or a compound disclosed in PCT Publication WO 2010/029082 for treating a disorder such as gastric cancer, breast cancer, pancreatic cancer, digestive cancer / gastrointestinal tract, solid tumor and head and neck cancer.

In one embodiment, the PI3K inhibitor or (S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2- yl)pyrrolidine-1,2-dicarboxamide (Compound A8) is administered at a dose of about 150-300, 200-300, 200-400 or 300-400 mg (e.g. per day), for example about 200, 300 or 400 mg.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a cytochrome P450 inhibitor (e.g., a CYP17 inhibitor) or a compound disclosed in PCT Publication WO 2010/149755 , for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the cytochrome P450 inhibitor (eg, CYP17 inhibitor) is a compound disclosed in PCT publication WO 2010/149755. In one embodiment, a PD-1 antibody molecule is used in combination with a compound disclosed in PCT publication WO 2010/149755 for the treatment of prostate cancer.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an HDM2 inhibitor, (S)-1-(4-chlorophenyl)-7-isopropoxy-6 -methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinoline-3( 4H)-one (Compound A10) or a compound disclosed in PCT Publication WO 2011/076786 for the treatment of a disorder, eg the disorder described herein). In one embodiment, the HDM2 inhibitor is (S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3 -oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one (Compound A10) or the compound disclosed in PCT publication WO 2011/076786. In one embodiment, the PD-1 antibody molecule is used in combination with (S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4- (4-methyl-3-oxopiperazin-1-yl)cyclohexyl)-methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one (Compound A10) or the compound disclosed in PCT Publication WO 2011/076786 , for the treatment of a disorder such as a solid tumor.

In one embodiment, an HDM2 inhibitor or (S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3 -oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one (Compound A10) is administered at a dose of approximately 400-700 mg, for example, administered three times a week, in for 2 weeks and a break for one week. In some embodiments, the dose is about 400, 500, 600, or 700 mg; about 400-500, 500-600 or 600-700 mg, for example, administered three times a week.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an iron-binding complexing agent, deferasirox (also known as Exjade; Compound A11), or a compound disclosed in PCT publication WO 1997/049395, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the iron-binding complexing agent is deferasirox or a compound disclosed in PCT publication WO 1997/049395. In one embodiment, the iron-binding complexing agent is deferasirox (Compound A11). In one embodiment, a PD-1 antibody molecule is used in combination with deferasirox (Compound A11) or a compound disclosed in PCT publication WO 1997/049395 to treat iron overload, hemochromatosis, or myelodysplasia.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with the aromatase inhibitor letrozole (also known as Femara; Compound A12) or a compound disclosed in US 4,978,672, for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the aromatase inhibitor is letrozole (Compound A12) or a compound disclosed in US Patent 4,978,672. In one embodiment, a PD-1 antibody molecule is used in combination with letrozole (Compound A12) or a compound disclosed in US Patent 4,978,672 to treat a disorder such as cancer, leiomyosarcoma, endometrial cancer, breast cancer, cancer of the female reproductive system, or deficit.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a PI3K inhibitor, e.g., a pan-PI3K inhibitor, (4S,5R)-3-(2 '-amino-2-morpholino-4'-(trifluoromethyl)-[4,5'-bipyrimidin]-6-yl)-4-(hydroxymethyl)-5-methyloxazolidin-2-one (Compound A13), or a compound, disclosed in PCT Publication WO2013/124826 for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PI3K inhibitor is (4S,5R)-3-(2'-amino-2-morpholino-4'-(trifluoromethyl)-[4,5'-bipyrimidin]-6-yl)-4-(hydroxymethyl )-5-methyloxazolidin-2-one (Compound A13) or the compound disclosed in PCT Publication WO2013/124826. In one embodiment, the PD-1 antibody molecule is used in combination with (4S,5R)-3-(2'-amino-2-morpholino-4'-(trifluoromethyl)-[4,5'-bipyrimidin]-6-yl )-4-(hydroxymethyl)-5-methyloxazolidin-2-one (Compound A13) or a compound disclosed in PCT Publication WO2013/124826 for the treatment of a disorder such as cancer or advanced solid tumor.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a p53 inhibitor and/or a p53/Mdm2 interaction inhibitor, (S)-5-(5- chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4-chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6 -dihydropyrrolo[3,4-d]imidazol-4(1H)-one (Compound A14), or a compound disclosed in PCT Publication WO2013/111105, for the treatment of a disorder, for example, the disorder described herein. In one embodiment, the p53 inhibitor and/or the p53/Mdm2 interaction inhibitor is (S)-5-(5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4- chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol-4(1H)-one (Compound A14) or the compound disclosed in PCT publication WO2013/111105. In one embodiment, the PD-1 antibody molecule is used in combination with (S)-5-(5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4-chlorophenyl) -2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol-4(1H)-one (Compound A14) or a compound disclosed in PCT Publication WO2013/111105, for the treatment of a disorder such as cancer or soft tissue sarcoma.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)- 2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, for the treatment of a disorder, e.g., the disorder described in this application. In one embodiment, the CSF-1R tyrosine kinase inhibitor is 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15) or a compound disclosed in PCT publication WO 2005/073224. In one embodiment, the PD-1 antibody molecule is used in combination with 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide ( Compound A15) or a compound disclosed in PCT Publication WO 2005/073224 for the treatment of a disorder such as cancer.

CSF-1-differentiated macrophages, sometimes referred to herein as M2 macrophages or tumor-associated macrophages (TAMs), when present in malignant tissues, are usually protumorogenic and can be characterized, for example, by expression of CSF-1R and CD163. In a cancerous tumor, CSF-1-differentiated macrophages often reflect compromised tissue integrity and/or an adaptive process initiated by tumors to maintain growth (Noy and Pollard (2014) Immunity; 41(1):49-61). To exert their protumorogenic activity, M2 macrophages secrete a variety of molecules such as cytokines, chemokines, growth factors, hormones, matrix-remodeling proteases and metabolites such as CSF-1 and CCL2, prostaglandin E2, and damage-associated molecular structures such as protein high motility groups box 1, extracellular adenosine triphosphate and degraded extracellular matrix components (Ruffell et al. (2012) Trends Immunol; 33(3):119-26; Zelenay et al. (2013) Trends Immunol; 34(7):329- 35). In vivo studies have shown that M2 macrophages also mediate resistance to chemotherapy by providing survival factors and/or activating anti-apoptotic programs in malignant cells. For example, neutralization of CSF-1 enhances response to chemotherapy in breast carcinomas (DeNardo et al. (2011) Cancer Discov. 1, 54-67). TAM macrophages are associated with vascular endothelial growth factor signaling and are regulators of tumor angiogenesis (Murdoch et al. (2008) Nat Rev Cancer; 8(8):618-31; Ruffell et al. (2012) Trends Immunol; 33(3): 119-26). M2 macrophages can also directly suppress CD8+ T cell proliferation in mouse tumor models (DeNardo et al. (2011) Cancer Discov. 1, 54-67; Doedens et al. (2010) Cancer Res; 70(19):7465- 75; Movahedi et al. (2010) Cancer Res; 70(14):5728-39; Ruffell et al. (2014) Cancer Cell; 26, 623-637). For example, macrophages directly downregulate T cell responses via PD-L1 expression in hepatocellular and ovarian carcinomas (Kuang et al. (2009) J Exp Med; 206(6):1327-37; Kryczek et al. (2006) J Exp Med ; 203(4):871-81).

Preclinical data indicate that TAM macrophages may serve as a therapeutic target, at least in part, as they are important drivers of various processes that contribute to tumor development, such as evading immune surveillance. For example, TAM differentiation, migration and survival can be regulated by CSF-1R when bound to soluble CSF. CSF-1R is a member of the growth factor receptor protein tyrosine kinase family, which includes several known proto-oncogenes (Ries et al. (2014) Cancer Cell 25(6):846-59). By disrupting the CSF-1R pathway, Compound A15 induced downregulation and reprogramming of M2-type macrophages in animal models of glioma, conferring antitumor efficacy through deletion/reprogramming of immunosuppressive TAMs. As a monotherapy, inhibition of CSF-1R alone prevents the growth of orthotopically implanted PDAC cell lines (Mitchem et al. (2013) Cancer Res; 73 (3): 1128-41) and causes regression of glioblastoma multiforme (GBM) (Pyontech et al. ( 2013) Nat Med; 19(10):1264-72).

Antitumor immunity in the tumor microenvironment can be suppressed by various tumor-infiltrating leukocytes, including Treg, MSC, and TAM. TAMs and MSCs can be present in tumors in large amounts, and their immunomodulatory activity often manifests itself locally in the tumor microenvironment. Without being limited by theory, it is believed that the mechanisms used by these cell types to suppress effective immunity include, for example, the secretion of cytokines such as IL-10 and TGF-β, and the expression of inhibitory receptors, for example, CTLA-4 and PD-L1. Removal of TAMs and MSCs, which are important in generative immunosuppression, provides, at least in part, the basis for combination strategies using checkpoint inhibitors, such as anti-PD-1 therapy, in those cancers in which TAMs contribute to tumor pathogenesis. Thus, a CSF-1R inhibitor can increase the clinical efficacy of a checkpoint inhibitor, such as an anti-PD-1 antibody molecule, and/or provide clinical efficacy, for example, in patients with tumors exhibiting high TAM expression (such as brain cancer (eg, glioblastoma), breast cancer (eg, TNBC), or pancreatic cancer), and in patients who do not respond effectively to an anti-PD-1 antibody molecule as the sole agent.

Without being limited by theory, it is believed that in some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)- N-methylpicolinamide (Compound A15), or the compound disclosed in PCT publication WO 2005/073224, selectively inhibits the activity of CSF-1R kinase. Compound A15 potently inhibits CSF-1R (IC ₅₀ 1.2 nM) as determined in an in vitro kinase activity assay with the recombinant CSF-1R kinase domain. The cellular activity of Compound A15 was demonstrated in reducing tyrosine-phosphorylated levels of CSF-1R in cells (EC ₅₀ pCSF-1R 58 nM) and a significant antiproliferative effect on the macrophage colony stimulating factor (M-CSF) dependent cell line MNFS-60 (EC ₅₀ 71 nM). The biochemical and cellular kinase selectivity of Compound A15 has been shown in a panel of more than 200 kinases, as well as at the cellular level, using enzyme-linked immunosorbent assays for autophosphorylation (ELISA) and kinase-transfected BaF3 proliferation assays. IC ₅₀ values for the respective subgroup of kinases, including related class III RTK kinases cKit and PDGFRβ, were at least >3 μM. The kinase selectivity of Compound A15 was confirmed at the cellular level as assessed by autophosphorylation assays (ECL ELISA assays) and BaF3 proliferation assays. Cellular EC _50s for pPDGFRβ, BAF3-PDGFRβ and pcKit were ≥2 μM, all other BaF3 cell lines showed >10 μM, while cell EC _50s for pCSF-1R and MCSF-dependent proliferation were <71 nM.

The biological effects of CSF-1R signaling include, for example, differentiation, proliferation, migration and/or survival of progenitor macrophages and monocytic lineage osteoclasts. Without being limited by theory, it is believed that in some embodiments, the mechanism by which Compound A15 mediates antitumor activity via CSF-1R signaling is to down-regulate or reprogram tumor-associated macrophages (TAMs) into classically activated phagocytic macrophages (Pyonteck et al. (2013) Nat Med 19(10):1264-72 Mao et al (2016) Clin Cancer Res Mar 8, DOI: 10.1158/1078-0432.CCR-15-1912, Epub ahead of print). Compound A15 (200 mg/kg daily) was shown to cause regression of established tumors and increase survival in a mouse genetic model of glioblastoma, and slow the growth of xenografts derived from proneural glioblastoma (Pyonteck et al. (2013) Nat Med; 19(10 ):1264-72). In a genetic model (regulated by mouse platelet growth factor B or PDG), Compound A15 efficacy was accompanied by a marked reprogramming from the M2 to M1 phenotype. The clinical potential of Compound A15 (200 mg/kg daily, for 10 days) has also been shown in an animal model of neuroblastoma (Mao et al. (2016) Clin Cancer Res. Mar 8, DOI: 10.1158/1078-0432.CCR-15 -1912). This study showed that CSF-1R+ infiltrating myeloid cells are suppressor cells and are indicative of a poor outcome in patients with neuroblastoma. The addition of Compound A15 restored the potential of myeloid cells to stimulate human T cell proliferation (Mao et al. (2016) Clin Cancer Res. Mar 8, DOI: 10.1158/1078-0432.CCR-15-1912). Similarly, primary human monocytes were co-cultured with neuroblastoma cell lines and were then shown to acquire potent suppressor activity against autologous T cells. Addition of Compound A15 provided inhibition of the suppressor function of tumor stimulated monocytes against CD8+ and CD4+ T cells (Mao et al. (2016) Clin Cancer Res. Mar 8, DOI: 10.1158/1078-0432.CCR-15-1912).

As described in Example 6, Compound A15 was also tested in combination with an exemplary anti-PD-1 antibody molecule in the MC38 syngeneic colorectal cancer model, selected for the presence of TAM macrophages and anti-PD-1 antibody response, and was it has been shown to inhibit tumor growth and increase survival compared to a single molecule of anti-PD-1 antibody.

In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered according to a periodic application schedule. Preclinical data indicate that comparable antitumor activity can be achieved with some intermittent regimens. Experiments on rats and monkeys showed an increase in transaminases with continuous daily use. These transaminase elevations are usually transient and a 7 day on/7 day off schedule may be chosen to obtain a tolerable profile and TAM depletion. In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered once a week. Preclinical data indicate that Compound A15 administration once a week (Q1W) provides at least the same efficacy as a daily dose in terms of TAM depletion and antitumor activity. In some embodiments, a dose of the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compounds A15), or a compound disclosed in PCT Publication WO, 2005/073224, when used in combination at least two (e.g., at least 3, 4, or 5) times lower than the highest tested dose level of a single CSF-tyrosine kinase inhibitor agent. 1R, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or the compound disclosed in the publication PCT WO 2005/073224. In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 50 mg to 1500 mg, e.g. 75 mg to 1000 mg, 100 mg to 900 mg, 200 mg to 800 mg, 300 mg up to 700 mg, 400 mg to 600 mg, 100 mg to 700 mg, 100 mg to 500 mg, 100 mg to 300 mg, 700 mg to 900 mg, 500 mg to 900 mg, 300 mg to 900 mg, 75 mg to 150 mg, 100 mg to 200 mg, 200 mg to 400 mg, 500 mg to 700 mg, or 800 mg to 1000 mg, e.g. at 50 mg, 100 mg, 150 mg 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, or mg, eg, daily (eg, according to a 7 day/7 day off schedule) or twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 100 mg, 150 mg, 300 mg, 600 mg or 900 mg. In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15) , or a compound disclosed in PCT publication WO 2005/073224, will be administered daily, for example, according to a 7 day/7 day break regimen, or once a week. In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT Publication WO 2005/073224, is administered at a dose of 50 mg to 150 mg, eg, about 100 mg, eg, daily (eg, according to a 7 day/7 day off schedule). In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 100 mg to 200 mg, eg about 150 mg, eg daily (eg 7 day/7 day off schedule). In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 200 mg to 400 mg, eg about 300 mg, eg daily (according to a 7 day/7 day off schedule). In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 500 mg to 700 mg, eg about 600 mg, eg daily (eg 7 day/7 day off schedule). In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 800 mg to 1000 mg, eg about 900 mg, eg daily (eg 7 day/7 day off schedule).

In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 50 mg to 150 mg, eg about 100 mg, once a week. In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 100 mg to 200 mg, eg about 150 mg, once a week. In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 200 mg to 400 mg, eg about 300 mg, once a week. In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15) , or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 500 mg to 700 mg, eg about 600 mg, once a week. In other embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15) , or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 800 mg to 1000 mg, eg about 900 mg, once a week.

In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered in combination with a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule). In some embodiments, a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (eg, a 400 mg dose), eg, once every 4 weeks. In other embodiments, the PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) is administered at a dose of 200 mg to 400 mg (eg, a 300 mg dose), eg, once every 3 weeks. In one embodiment, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 50 mg to 150 mg (e.g., about 100 mg), e.g. once a week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg), for example, once every 4 weeks, or at a dose of 200 mg to 400 mg (for example, at a dose of 300 mg), for example, once every 3 weeks, for example, by intravenous infusion. In another embodiment, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 100 mg to 200 mg (e.g., about 150 mg), e.g. per week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg), for example, once every 4 weeks, or in dose of 200 mg to 400 mg (eg 300 mg dose), eg once every 3 weeks, eg by intravenous infusion. In another embodiment, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15) , or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 200 mg to 400 mg (e.g., about 300 mg), e.g. week, for example, orally, and the PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg), for example, once every 4 weeks, or at a dose 200 mg to 400 mg (eg at a dose of 300 mg), eg once every 3 weeks, eg by intravenous infusion. In another embodiment, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose of 500 mg to 700 mg (e.g., about 600 mg), e.g. once a week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg), for example, once every 4 weeks, or at a dose of 200 mg to 400 mg (for example, at a dose of 300 mg), for example, once every 3 weeks, for example, by intravenous infusion. In another embodiment, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)-amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15 ), or a compound disclosed in PCT publication WO 2005/073224, is administered at a dose between 800 mg and 1000 mg (e.g., about 900 mg), e.g. per week, for example, orally, and a PD-1 inhibitor (for example, an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg), for example, once every 4 weeks, or in dose of 200 mg to 400 mg (eg 300 mg dose), eg once every 3 weeks, eg by intravenous infusion.

In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered orally. In some embodiments, the anti-PD-1 antibody molecule is administered intravenously. In some embodiments, the anti-PD-1 antibody molecule is administered by intravenous infusion, e.g., over 15 minutes to 3 hours, e.g., over 30 minutes to 2 hours, e.g., over 30 minutes, 1 hour, 1.5 hours or 2 hours. In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15) , or a compound disclosed in PCT publication WO 2005/073224, is administered prior to administration of the anti-PD-1 antibody molecule, eg, on an empty stomach. For example, administration of an anti-PD-1 antibody molecule may begin any time after administration of the tyrosine kinase inhibitor CSF-1R, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazole-6- yl)oxy)-N-methylpicolinamide (Compounds A15), or the compound disclosed in PCT Publication WO 2005/073224.

In some embodiments, the CSF-1R tyrosine kinase inhibitor, 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide (Compound A15), or a compound disclosed in PCT publication WO 2005/073224, is administered in combination with a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule) to treat cancer, eg, a solid tumor (eg, an advanced solid tumor). Examples of cancers that can be treated with the combination include, but are not limited to, brain cancer (eg, glioblastoma multiforme (GBM), eg, recurrent glioblastoma), breast cancer (eg, triple-negative breast cancer (eg, TNBC)) or pancreatic cancer (eg, advanced pancreatic cancer). Common features of these cancers include, for example, tumor biology characterized by high levels of TAM in the tumor microenvironment, which may contribute to immune evasion and immune suppression. In some embodiments, blockade of CSF-1R in combination with anti-PD-1 therapy may, for example, cause TAM reprogramming and/or immune suppression of tumor infiltrating lymphocytes (TILs).

In another embodiment, the combination, for example, a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an apoptosis inducer and/or angiogenesis inhibitor such as imatinib mesylate (also known as Glivec®; Compound A16), or a compound disclosed in PCT Publication WO1999/003854, for the treatment of a disorder, eg the disorder described. In one embodiment, the apoptosis inducer and/or angiogenesis inhibitor is imatinib mesylate (Compound A16) or a compound disclosed in PCT Publication WO1999/003854. In one embodiment, a PD-1 antibody molecule is used in combination with imatinib mesylate (Compound A16) or a compound disclosed in PCT Publication WO1999/003854 to treat a disorder such as cancer, multiple myeloma, prostate cancer, non-small cell lung cancer, lymphoma , gastric cancer, melanoma, breast cancer, pancreatic cancer, gastrointestinal/gastrointestinal tract cancer, colon and rectal cancer, glioblastoma multiforme, liver cancer, head and neck cancer, asthma, multiple sclerosis, allergies, Alzheimer's dementia , amyotrophic lateral sclerosis or rheumatoid arthritis.

In some embodiments, imatinib mesylate (Compound A16) is administered at a dose of about 100-1000 mg, such as about 200 mg to 800 mg, about 300 mg to 700 mg, or about 400 mg to 600 mg, such as about 200 mg , 300 mg, 400 mg, 500 mg, 600 mg or 700 mg. The administration schedule may vary, for example, from once every two days to once a day, two or three times a day. In one embodiment, imatinib mesylate is administered daily at an oral dose of about 100 mg to about 600 mg, such as about 100 mg, 200 mg, 260 mg, 300 mg, 400 mg, or 600 mg daily.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a JAK inhibitor, 2-fluoro-N-methyl-4-(7-(quinoline-6 -ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide (Compound A17), or its dihydrochloride, or a compound disclosed in PCT publication WO 2007/070514, for the treatment of a disorder, for example, the violation described in this application. In one embodiment, the JAK inhibitor is 2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide ( Compound A17) or its dihydrochloride, or the compound disclosed in PCT publication WO 2007/070514. In one embodiment, the PD-1 antibody molecule is used in combination with 2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin- 2-yl)benzamide (Compound A17) or dihydrochloride thereof, or a compound disclosed in PCT publication WO 2007/070514, for the treatment of a disorder such as colon and rectal cancer, myeloid leukemia, hemoblastosis, autoimmune disease, non-Hodgkin's lymphoma or thrombocythemia.

In one embodiment, a JAK inhibitor or 2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide ( Compound A17), or its dihydrochloride, is administered at a dose of about 400-600 mg (eg per day), for example about 400, 500 or 600 mg, or about 400-500 or 500-600 mg.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a JAK inhibitor, ruxolitinib phosphate (also known as Jakavi; Compound A18), or a compound disclosed in PCT publication WO 2007/070514, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the JAK inhibitor is ruxolitinib phosphate (Compound A18) or a compound disclosed in PCT publication WO 2007/070514. In one embodiment, a PD-1 antibody molecule is used in combination with ruxolitinib phosphate (Compound A18) or a compound disclosed in PCT publication WO 2007/070514 to treat a disorder such as prostate cancer, lymphocytic leukemia, multiple myeloma, lymphoma, cancer lung, leukemia, cachexia, breast cancer, pancreatic cancer, rheumatoid arthritis, psoriasis, colon and rectal cancer, myeloid leukemia, hemoblastosis, autoimmune disease, non-Hodgkin's lymphoma, or thrombocythemia.

In one embodiment, the JAK inhibitor or ruxolitinib phosphate (Compound A18) is administered at a dose of about 15-25 mg, eg, twice daily. In some embodiments, the dosage is about 15, 20, or 25 mg, or about 15-20 or 20-25 mg.

In another embodiment, the combination, for example, a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a deacetylase (DAC) inhibitor panobinostat (Compound A19) or a compound disclosed in PCT publication WO 2014/ 072493, for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the DAC inhibitor is panobinostat (Compound A19) or a compound disclosed in PCT publication WO 2014/072493. In one embodiment, a PD-1 antibody molecule is used in combination with panobinostat (Compound A19), a compound disclosed in PCT publication WO 2014/072493, for the treatment of a disorder such as colon and rectal cancer, small cell lung cancer, respiratory tract cancer/ breast cancer, prostate cancer, multiple myeloma, myelodysplastic syndrome, bone cancer, non-small cell lung cancer, endocrine cancer, lymphoma, neurological cancer, leukemia, HIV/AIDS, immunological disorder, transplant rejection, gastric cancer, melanoma, breast cancer (eg, triple-negative breast cancer (TNBC)), pancreatic cancer, colon and rectal cancer, glioblastoma multiforme, myeloid leukemia, hemoblastosis, kidney cancer, non-Hodgkin's lymphoma, head and neck cancer, hematopoietic disorder, or liver cancer. In some embodiments, the cancer is selected from colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS CRC), lung cancer (e.g., non-small cell lung cancer), or breast cancer (e.g., triple negative lung cancer (TNBC) ).

In some embodiments, the deacetylase (DAC) inhibitor, panobinostat (Compound A19), or the compound disclosed in PCT publication WO 2014/072493, is a histone deacetylase (HDAC) inhibitor. Without being limited by theory, it is believed that in some embodiments, the deacetylase (DAC) inhibitor, panobinostat (Compound A19), or the compound disclosed in PCT publication WO 2014/072493, belongs to the class of compounds of cinnamic-hydroxamic acid derivatives and is a pan histone inhibitor. (and nonhistone) DAC (HDAC) I, II and IV class. Such HDACs are epigenetic modulators and important cancer targets due to the dysregulation of these enzymes in many tumor types. DAC enzymes also specifically interact with lysine groups on various nonhistone proteins such as p53, α-tubulin, Hsp90, and HIF1-α. Thus, in some embodiments, the deacetylase (DAC) inhibitor, panobinostat (Compound A19), or the compound disclosed in PCT publication WO 2014/072493, is a pan DAC inhibitor. A deacetylase (DAC) inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO 2014/072493, can provide a multifaceted approach to inhibition of proliferation and/or survival of cancer cells, for example, through its effects on histone acetylation, gene expression and /or oncogenic function of nonhistone proteins such as Hsp90. Panobinostat (Compound A19), for example, as a pan HDAC inhibitor, is very effective in inhibiting HDAC activity of most class I, IIa, IIb and IV isoforms at low nanomolar concentrations.

In some embodiments, the combination described herein includes a deacetylase (DAC) inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO 2014/072493, and an inhibitor of an immune checkpoint molecule, such as a PD-1 inhibitor ( for example, an anti-PD-1 antibody molecule).

Without being limited by theory, it is believed that in some embodiments, epigenetic modifiers, such as HDAC inhibitors, can be used to enhance immunotherapy. Panobinostat (Compound A19) upregulated PD-L1 gene expression in melanoma cell lines, cultured melanoma biopsy material, and tumor-bearing mice. Panobinostat (Compound A19) also modestly improved responses to PD-1 inhibitor treatment in mice (Woods et al. (2015) Cancer Immunol. Res. p. 1375-85). Clinical data indicate that epigenetic modifiers, including the combination of an HDAC inhibitor with a DNMT inhibitor, are active in some patients with NSCLC (Juergens et al. (2011) Cancer Discov. p. 598-607). Preclinical data indicate that immunotherapies such as PD-1 or CTLA-4 inhibitors can enhance this activity (Kim et al. (2014) Proc. Natl. Acad. Sci. U.S.A. p. 11774-9). In some embodiments, the deacetylase (DAC) inhibitor panobinostat (Compound A19) or the compound disclosed in PCT publication WO 2014/072493 enhances or is used to enhance immunotherapy, e.g., immunotherapy comprising a PD-1 inhibitor (e.g., an anti-PD- 1). In some embodiments, a deacetylase (DAC) inhibitor panobinostat (Compound A19) or a compound disclosed in PCT publication WO 2014/072493 and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), administered at a dose and/or time schedule that, in combination, provide the desired antitumor activity.

In some embodiments, the approved dose of panobinostat (Compound A19) for multiple myeloma is 20 mg biday, three times a week (TIW), with panobinostat (Compound A19) administered for two weeks and off for a week over a 21-day course. In other embodiments, a less frequent one week/one week off regimen over a 28 day course was also tested in multiple myeloma patients using a TIW dose of 30-60 mg. In some embodiments, the deacetylase (DAC) inhibitor panobinostat (Compound A19) or the compound disclosed in PCT publication WO 2014/072493 is administered at a dose of TIW 10 mg on a one week/one week off schedule, e.g., in combination with a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule). The dose and regimen may be increased to the TIW 20 mg regimen given two weeks/one week off, if possible, depending on the safety and tolerability of the lower dose and the less frequent regimen. If the dose of TIW 10 mg on a one week/one week off schedule is unacceptably toxic, a lower dose or less frequent dosing regimen may be used.

In one embodiment, the DAC inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO, 2014/072493, is administered at a dose of approximately 20 mg (eg per day).

In one embodiment, a DAC inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO 2014/072493, is administered at a dose of 2 mg to 30 mg, 5 mg to 20 mg, or 10 mg to 15 mg, e.g. , 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 30 mg e.g. once a day, three times a week, or once a week, e.g. one week/break schedule for one week, for one week/break for three weeks, or for two weeks/break for one week. In one embodiment, the DAC inhibitor or panobinostat (Compound A19) is administered at a dose of 5 mg to 15 mg, e.g., about 10 mg, e.g., three times per week, e.g. within one week/break for one week. In another embodiment, the DAC inhibitor or panobinostat (Compound A19) is administered at a dose of 15 mg to 25 mg, such as 20 mg, such as three times per week, such as a two-week/three-week break schedule.

In some embodiments, the DAC inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO 2014/072493, is administered orally.

In one embodiment, a DAC inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO 2014/072493, is administered at a dose of 5 mg to 15 mg (e.g., 10 mg), e.g., three times per week ( e.g., one week/three weeks off or two weeks/one week off), e.g., orally, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (for example, at a dose of 400 mg), for example, once every 4 weeks, for example, by intravenous infusion.

In one embodiment, a DAC inhibitor, panobinostat (Compound A19), or a compound disclosed in PCT publication WO 2014/072493, is administered in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), for the treatment of colon and rectum cancer. colon (eg, MSS CRC), lung cancer (eg, non-small cell lung cancer (NSCLC), or breast cancer (eg, triple negative breast cancer (TNBC)).

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an inhibitor of one or more of cytochrome P450 (e.g., 11B2), aldosterone, or angiogenesis, osilodrostat (Compound A20), or a compound disclosed in PCT Publication WO2007/024945, for the treatment of a disorder, eg the disorder described herein. In one embodiment, the inhibitor of one or more of cytochrome P450 (eg, 11B2), aldosterone, or angiogenesis is osilodrostat (Compound A20) or a compound disclosed in PCT Publication WO2007/024945. In one embodiment, a PD-1 antibody molecule is used in combination with osilodrostat (Compound A20) or a compound disclosed in PCT Publication WO2007/024945 for the treatment of a disorder such as Cushing's syndrome, hypertension or heart failure therapy.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-( S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or a compound disclosed in US 8,552,003, for treatment of a disorder, for example, a disorder described in this application. In one embodiment, the IAP inhibitor is (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Patent 8,552,003. In one embodiment, the PD-1 antibody molecule is used in combination with (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidine -1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Pat. No. 8,552,003 for the treatment of a disorder such as multiple myeloma, colon and rectal cancer (CLC), lung cancer (e.g., non-small cell lung cancer (NSCLC), breast cancer (e.g., triple negative breast cancer (TNBC)), ovarian cancer, pancreatic cancer, or hematopoietic disorder. In some embodiments, the cancer is selected from colon and rectal cancer (e.g., , microsatellite stable colon and rectal cancer (MSS CRC), lung cancer (eg, non-small cell lung cancer), or breast cancer (eg, triple negative lung cancer (TNBC)).

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Pat. No. 8,552,003 is a low molecular weight orally available SMAC mimetic that binds to the BIR3 domain of CIAP1, XIAP and optionally CIAP2. The CIAP1 and CIAP2 proteins are components of protein complexes of the TNF receptor family containing death domains. Without being limited by theory, it is believed that in some embodiments, the binding of the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl )pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. downstream NF-κB signaling after receptors (Gyrd-Hansen M, Meier P (2010) Nat. Rev. Cancer p. 561-74). In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is used to enhance antitumor immunity in a subject. For example, an IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2- oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, can increase the proliferation and function of human and murine T cells in vitro after costimulation and/or response to prophylactic and therapeutic antitumor vaccines in vivo eg via NF-κB activation (Dougan et al. (2010) J. Exp. Med. p. 2195-206). NF-κB activity has been implicated in T cell cross-priming during immunogenic cell apoptosis in response to the release of risk-associated inflammatory molecules (Yatim et al. (2015) Science p. 328-34), suggesting a mechanism for enhancing T cell function and antitumor immunity. As another example, an IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl) -2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Patent 8,552,003, can protect monocytic dendritic cells from apoptosis similar to CD40 ligation (Knights et al. (2013) Cancer Immunol. Immunother. p 321-35).

In some embodiments, the combination described herein includes an IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl )pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or a compound disclosed in US Pat. No. 8,552,003, and an immune checkpoint molecule inhibitor, e.g. antibodies against PD-1).

Without being limited by theory, it is believed that in some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl) pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is an orally available small molecule that activates NF-kB signaling after members of the TNF receptor family . NF-κB is a major transcriptional regulator in immune cells and also acts directly in tumor cells (Perkins (2012) Nat. Rev. Cancer p. 121-32). Preclinical data indicate that NF-κB activity is required for cross-priming of CD8+ T lymphocytes undergoing immunogenic apoptosis (Yatim et al. (2015) Science p. 328-34). Compound A21 stimulated T-lymphocyte proliferation, induced IFN-gamma, and suppressed IL-10 production in vitro . Clinical studies with Compound A21 have demonstrated the induction of circulating cytokines including TNF alpha, IL-8, IL-10 and CCL2 (Infante et al. (2014) J. Clin. Oncol. p. 3103-10). Clinical data indicate that Compound A21 increased the percentage of patients with TNBC with a complete response in the laboratory study when treated with paclitaxel. In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, increases or is used to increase the antitumor activity of a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule).

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or a compound disclosed in US Pat. No. 8,552,003, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), are administered at a dose and/or timing schedule that, in combination, provide the desired antitumor activity.

Without being limited by theory, doses of the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1 -yl)-2-oxoethyl)-2-(methylamino)propanamide (Compounds A21) or the compound disclosed in US Pat. For example, in paired skin biopsies of two patients receiving a 320 mg dose, both showed degradation of CIAP1, the target protein of Compound A21; skin biopsies collected from patients receiving lower doses (160 mg and below) showed less pronounced pharmacodynamic activity (Infante et al. (2014) J. Clin. Oncol. p. 3103-10). In some embodiments, doses above 900 mg are not required for clinical efficacy. In some situations, the 1800 mg dose of Compound A21 was poorly tolerated when used in combination with paclitaxel. Thus, in one embodiment, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1- yl)-2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of 300 mg to 900 mg, for example, once a week. In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered less frequently than once a week, e.g. at four weeks.

In one embodiment, an IAP inhibitor or (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US 8,552,003 is administered at a dose of about 1800 mg or less, for example once a week.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. up to about 900 mg, from 400 mg to about 800 mg, or from about 500 mg to about 700 mg, for example, at a dose of about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, or approximately 900 mg, for example, once a week, once every two weeks, once every three weeks, or once every four weeks.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of about 200 mg to about 400 mg, such as about 300 mg, once a week. In other embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of about 800 mg to about 1000 mg, such as about 900 mg, once a week. In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered at a dose of about 200 mg to about 400 mg, such as about 300 mg, once every four weeks.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered orally.

In some embodiments, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21), or the compound disclosed in US Pat. No. 8,552,003, is administered in combination with a PD-1 inhibitor (eg, an anti-PD-1 antibody molecule). In one embodiment, the IAP inhibitor, (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)- 2-oxoethyl)-2-(methylamino)propanamide (Compound A21) or the compound disclosed in US Pat. No. 8,552,003 is administered at a dose of 200 mg to 400 mg (e.g. 300 mg), e.g. once a week, e.g., orally, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg dose), e.g., once every 4 weeks, e.g., by intravenous infusion .

In some embodiments, an IAP inhibitor, such as (S)-N-(S)-1-cyclohexyl-2-(S)-2-(4-(4-fluorobenzoyl)-thiazol-2-yl)pyrrolidin-1- yl)-2-oxoethyl)-2-(methylamino)propane-amide (Compound A21) or the compound disclosed in US Pat. No. 8,552,003 is administered in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) colon and rectal cancer (eg, MSS CRC), lung cancer (eg, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple-negative breast cancer (eg, TNBC)).

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with the Smoothened (SMO) inhibitor, sonidegib phosphate (Compound A22), (R)-2-( 5-(4-(6-benzyl-4,5-dimethylpyridazin-3-yl)-2-methylpiperazin-1-yl)pyrazin-2-yl)propan-2-ol (Compound A25) or a compound disclosed in PCT Publication WO 2007/131201 or WO 2010/007120, for the treatment of a disorder, for example the disorder described herein. In one embodiment, the SMO inhibitor is sonidegiba phosphate (Compound A22), (R)-2-(5-(4-(6-benzyl-4,5-dimethylpyridazin-3-yl)-2-methylpiperazin-1-yl) pyrazin-2-yl)propan-2-ol (Compound A25) or a compound disclosed in PCT publication WO 2007/131201 or WO 2010/007120. In one embodiment, the PD-1 antibody molecule is used in combination with sonidegib phosphate (Compound A22), (R)-2-(5-(4-(6-benzyl-4,5-dimethylpyridazin-3-yl)-2- methylpiperazin-1-yl)pyrazin-2-yl)propan-2-ol (Compound A25) or a compound disclosed in PCT Publication WO 2007/131201 or WO 2010/007120 for the treatment of a disorder such as cancer, medulloblastoma, small cell carcinoma lung, prostate cancer, basal cell carcinoma, pancreatic cancer, or inflammation.

In some embodiments, sonidegib phosphate (Compound A22) is administered at a dose of about 20-500 mg, such as about 40 mg to 400 mg, about 50 mg to 300 mg, or about 100 mg to 200 mg, such as about 50 mg , 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. The administration schedule may vary, for example, from once every two days to once a day, two or three times a day.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an Alk inhibitor, ceritinib (also known as Zykadia; Compound A23), or a compound disclosed in the publication PCT WO 2007/131201, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the Alk inhibitor is ceritinib (Compound A23) or a compound disclosed in PCT publication WO 2007/131201. In one embodiment, a PD-1 antibody molecule is used in combination with ceritinib (Compound A23) or a compound disclosed in PCT publication WO 2007/131201 to treat a disorder such as non-small cell lung cancer or solid tumors.

In one embodiment, the Alk inhibitor or ceritinib (Compound A23) is administered at a dose of about 750 mg, eg, once a day.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a JAK and/or CDK4/6 inhibitor, 7-cyclopentyl-N,N-dimethyl-2 -((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide (Compound A24), or the compound disclosed in US Patent 8,415,355 or US patent 8,685,980, for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the JAK and/or CDK4/6 inhibitor is 7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[ 2,3-d]pyrimidine-6-carboxamide (Compound A24) or a compound disclosed in US 8,415,355 or US 8,685,980. In one embodiment, the PD-1 antibody molecule is used in combination with 7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2 ,3-d]pyrimidine-6-carboxamide (Compound A24) or a compound disclosed in US 8,415,355 or US 8,685,980 for the treatment of a disorder such as lymphoma, neurological cancer, melanoma, breast cancer or solid tumor.

In one embodiment, a JAK and/or CDK4/6 inhibitor or 7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2 ,3-d]pyrimidine-6-carboxamide (Compound A24) is administered at a dose of about 200-600 mg, for example, per day. In one embodiment, the compound is administered at a dose of about 200, 300, 400, 500, or 600 mg, or about 200-300, 300-400, 400-500, or 500-600 mg.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a prolactin receptor (PRLR) inhibitor, a human monoclonal antibody molecule (Compound A26) disclosed in US Pat. 7,867,493, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the PRLR inhibitor is a human monoclonal antibody (Compound A26) disclosed in US 7,867,493. In one embodiment, a PD-1 antibody molecule is used in combination with a human monoclonal antibody molecule (Compound A26) described in US Pat. No. 7,867,493 to treat a disorder such as cancer, prostate cancer, or breast cancer.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a PIM kinase inhibitor, N-(4-((1R, 3S, 5S)-3- amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide (Compound A27), or a compound disclosed in PCT publication WO 2010/026124, for the treatment of a disorder, e.g. described in this application. In one embodiment, the PIM kinase inhibitor is N-(4-((1R,3S,5S)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide (Compound A27) or a compound disclosed in PCT publication WO 2010/026124. In one embodiment, the PD-1 antibody molecule is used in combination with N-(4-((1R,3S,5S)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl )-5-fluoropicolinamide (Compound A27) or a compound disclosed in PCT publication WO 2010/026124 for the treatment of a disorder such as multiple myeloma, myelodysplastic syndrome, myeloid leukemia or non-Hodgkin's lymphoma.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a Wnt signaling inhibitor, 2-(2',3-dimethyl-[2,4'- bipyridine]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide (Compound A28), or a compound disclosed in PCT publication WO 2010/101849, for the treatment of a disorder, for example, violation described in this application. In one embodiment, the Wnt signaling inhibitor is 2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl) acetamide (Compound A28) or a compound disclosed in PCT publication WO 2010/101849. In one embodiment, the Wnt signaling inhibitor is 2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl) acetamide (Compound A28). In one embodiment, the PD-1 antibody molecule is used in combination with 2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridine -2-yl)acetamide (Compound A28) or a compound disclosed in PCT Publication WO 2010/101849 for the treatment of a disorder such as a solid tumor (e.g. head and neck cancer, squamous cell carcinoma, breast cancer, pancreatic cancer or cancer large intestine). In some embodiments, the cancer is selected from skin cancer (eg, melanoma), high microsatellite instability (MSI-H) solid tumor, pancreatic cancer, or breast cancer (eg, triple negative breast cancer (TNBC)).

In some embodiments, 2-(2',3-dimethyl-[2,4'-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide (Compound A28 ) is administered at a dose of about 1-50 mg, such as about 2 mg to 45 mg, about 3 mg to 40 mg, about 5 mg to 35 mg, 5 mg to 10 mg, or about 10 mg to 30 mg eg about 2 mg, 5 mg, 10 mg, 20 mg, 30 mg, or 40 mg. The administration schedule may vary, for example, from once every two days to once a day, two or three times a day.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a BRAF inhibitor, encorafenib (Compound A29), or a compound disclosed in PCT Publication WO 2011/025927 , for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the BRAF inhibitor is encorafenib (Compound A29) or a compound disclosed in PCT publication WO 2011/025927. In one embodiment, a PD-1 antibody molecule is used in combination with encorafenib (Compound A29) or a compound disclosed in PCT publication WO 2011/025927 to treat a disorder such as non-small cell lung cancer, melanoma, or colon and rectal cancer.

In one embodiment, the BRAF inhibitor or encorafenib (Compound A29) is administered at a dose of about 200-300, 200-400, or 300-400 mg, for example, per day. In one embodiment, the compound is administered at a dose of about 200, about 300, or about 400 mg.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a CDK4/6 inhibitor, 7-cyclopentyl-N,N-dimethyl-2-((5 -((1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo[4.2.1]nonan-3-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d ]pyrimidine-6-carboxamide (Compound A30), or a compound disclosed in PCT publication WO 2011/101409, for the treatment of a disorder, for example, the disorder described herein. In one embodiment, the CDK4/6 inhibitor is 7-cyclopentyl-N,N-dimethyl-2-((5-((1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo[4.2.1] no-nan-3-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide (Compound A30) or the compound disclosed in PCT Publication WO 2011/101409. In one embodiment, the PD-1 antibody molecule is used in combination with 7-cyclopentyl-N,N-dimethyl-2-((5-((1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo[ 4.2.1]nonan-3-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide (Compound A30) or a compound disclosed in PCT publication WO 2011/101409, for the treatment of a disorder such as cancer, mantle cell lymphoma, liposarcoma, non-small cell lung cancer, melanoma, squamous cell carcinoma of the esophagus, or breast cancer.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a HER3 inhibitor, Compound A31, or a compound disclosed in PCT publication WO 2012/022814, for the treatment violations, for example, the violations described in this application. In one embodiment, the HER3 inhibitor is Compound A31 or a compound disclosed in PCT publication WO 2012/022814. In one embodiment, a PD-1 antibody molecule is used in combination with Compound A31 or a compound disclosed in PCT Publication WO 2012/022814 to treat a disorder such as gastric cancer, esophageal cancer, head and neck cancer, squamous cell carcinoma, gastric cancer, breast cancer (eg, metastatic breast cancer) or cancer of the digestive/gastrointestinal tract.

In some embodiments, Compound A31 is a human monoclonal antibody molecule.

In one embodiment, the HER3 inhibitor or Compound A31 is administered at a dose of about 3, 10, 20, or 40 mg/kg, eg, once a week (QW). In one embodiment, the compound is administered at a dose of about 3-10, 10-20, or 20-40 mg/kg.

In another embodiment, the combination, for example, a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an FGFR2 and/or FGFR4 inhibitor, Compound A32, or a compound disclosed in PCT publication WO 2014/ 160160 (eg, an anti-FGFR2 and/or FGFR4 antibody-drug molecule conjugate, eg, mAb 12425), for the treatment of a disorder, eg, the disorder described herein. In one embodiment, the FGFR2 and/or FGFR4 inhibitor is Compound A32 or a compound disclosed in PCT Publication WO 2014/160160. In one embodiment, the PD-1 antibody molecule is used in combination with Compound A32 or the compound described in Table 7 to treat a disorder such as cancer, gastric cancer, breast cancer, rhabdomyosarcoma, liver cancer, adrenal cancer, lung cancer, cancer esophagus, colon cancer, or endometrial cancer.

In some embodiments, Compound A32 is a conjugate of an anti-FGFR2 and/or FGFR4 antibody-drug molecule, such as mAb 12425.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an M-CSF inhibitor, Compound A33, or a compound disclosed in PCT Publication WO 2004/045532 ( for example, an antibody molecule or a Fab fragment against M-CSF), for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the M-CSF inhibitor is Compound A33 or a compound disclosed in PCT publication WO 2004/045532. In one embodiment, a PD-1 antibody molecule is used in combination with Compound A33 or a compound described in PCT publication WO 2004/045532 to treat a disorder such as cancer, prostate cancer, breast cancer, or pigmented villonodular synovitis (PVNS).

In embodiments, Compound A33 is an anti-M-CSF monoclonal antibody molecule or a fragment (eg, a Fab fragment) thereof. In embodiments, the M-CSF inhibitor or Compound A33 is administered at an average dose of about 10 mg/kg.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a MEK inhibitor, binimetinib (Compound A34), or a compound disclosed in PCT Publication WO 2003/077914 , for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the MEK inhibitor is binimetinib (Compound A34) or a compound disclosed in PCT publication WO 2003/077914. In one embodiment, a PD-1 antibody molecule is used in combination with binimetinib (Compound A34) or a compound disclosed in PCT publication WO 2003/077914 to treat a disorder such as non-small cell lung cancer, multisystem genetic disorder, melanoma, ovarian cancer, cancer digestive/gastrointestinal tract, rheumatoid arthritis, or colon and rectal cancer.

In one embodiment, the MEK inhibitor or binimetinib (Compound A34) is administered at a dose of about 45 mg, eg, twice daily.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an inhibitor of one or more of c-KIT, histamine release, Flt3 (e.g., FLK2/STK1) or PKC, midostaurin (Compound A35), or a compound disclosed in PCT publication WO 2003/037347 for the treatment of a disorder, for example, the disorder described herein. In one embodiment, the inhibitor is midostaurin (Compound A35) or a compound disclosed in PCT publication WO 2003/037347. In one embodiment, the inhibitor of one or more of c-KIT, histamine release, Flt3 (eg, FLK2/STK1), or PKC is midostaurin. In one embodiment, a PD-1 antibody molecule is used in combination with midostaurin (Compound A35) or a compound disclosed in PCT publication WO 2003/037347 to treat a disorder such as cancer, colon and rectal cancer, myeloid leukemia, myelodysplastic syndrome, age-related macular degeneration, a complication of diabetes, or a dermatological disorder.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a TOR inhibitor (e.g., an mTOR inhibitor), everolimus (also known as Afinitor; Compound A36), or a Compound disclosed in PCT Publication WO 2014/085318 for the treatment of a disorder, such as the disorder described herein). In one embodiment, the TOR inhibitor is everolimus (Compound A36) or the Compound disclosed in PCT publication WO 2014/085318. In one embodiment, a PD-1 antibody molecule is used in combination with everolimus (Compound A36) to treat a disorder such as colon and rectal cancer, interstitial lung disease, small cell lung cancer, respiratory tract/sternum cancer, prostate cancer, multiple myeloma , sarcoma, age-related macular degeneration, bone cancer, tuberous sclerosis, non-small cell lung cancer, endocrine cancer, lymphoma, neurological disorders, astrocytoma, cervical cancer, neurological cancer, leukemia, immune disorders, transplant rejection, gastric cancer, melanoma, epilepsy, breast cancer cancer (eg, triple-negative breast cancer (TNBC)) or bladder cancer. In some embodiments, the cancer is selected from colon and rectal cancer (e.g., microsatellite stable colon and rectal cancer (MSS CRC), lung cancer (e.g., non-small cell lung cancer), or breast cancer (e.g., triple negative breast cancer (TNBC). )).

In some embodiments, an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, inhibits the mammalian target of rapamycin (mTOR), a serine/threonine kinase downstream of the PI3K/AKT pathway. The mTOR pathway is deregulated in several human cancers. Without being limited by theory, it is believed that in some embodiments, the mTOR inhibitor, everolimus (Compound A36), or the compound disclosed in PCT publication WO 2014/085318, binds to the intracellular protein FKBP-12, resulting in the formation of an inhibitory complex with mTOR complex 1 (mTORC1) and thus inhibition of mTOR kinase activity. Everolimus (Compound A36) may decrease the activity of S6 ribosomal protein kinase (S6K1) and/or eukaryotic initiation factor 4E binding protein (4E-BP1), downstream mTOR effectors involved in protein synthesis. S6K1 is a substrate of mTORC1 and phosphorylates activation domain 1 of the estrogen receptor, resulting in ligand-independent activation of the receptor. In addition, everolimus (Compound A36) can inhibit the expression of hypoxia inducible factor (eg, HIF-1) and/or reduce the expression of vascular endothelial growth factor (VEGF). Inhibition of mTOR by everolimus (Compound A36) has been shown to reduce cell proliferation, angiogenesis, and glucose uptake in in vitro and/or in vivo studies . Constitutive activation of the PI3K/Akt/mTOR pathway may contribute to endocrine resistance in breast cancer. In vitro studies show that estrogen-dependent and HER2+ breast cancer cells are sensitive to the inhibitory effects of everolimus (Compound A36), and that combined treatment with everolimus and Akt, HER2 or aromatase inhibitors enhances the antitumor activity of everolimus (Compound A36) in a synergistic manner. Two regulators of mTORC1 signaling are oncogene suppressors, tuberin sclerosis complexes 1 and 2 (TSC1, TSC2). Loss or inactivation of TSC1 or TSC2 results in the activation of downstream signaling. In TSC, a genetic disorder, inactivating mutations in the TSC1 or TSC2 gene result in the formation of hamartomas throughout the body.

In some embodiments, the combination described herein includes an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., a molecule antibodies against PD-1).

Everolimus (Compound A36) has been used clinically as a direct anticancer agent at a dose of 10 mg daily to suppress mTOR activity in tumor cells, and as an immunosuppressant in patients requiring solid organ transplantation at a dose of 1.5-2.0 mg daily to suppress the function of T-lymphocytes. Without being limited by theory, it is believed that in some embodiments, at such dosages and schedules, everolimus (Compound A36) is expected to attenuate an effective antitumor response. Immunoaging is a decline in immune function that occurs in the elderly and includes a reduced response to vaccination, including influenza vaccination. The decline in immune function with age includes an increase in PD-1 positive "depleted" T lymphocytes, which show a reduced response to antigen challenge (Lages et al. (2010) Aging Cell 9, 785-798). Clinical data indicate that a dose of 5 mg per week may be immunostimulatory, reducing the percentage of PD-1-positive CD4+ and CD8+ lymphocytes compared with placebo treatment and enhancing influenza vaccine response in elderly subjects (Mannick et al. (2014) Sci. Transl. Med. Vol. 6, Issue 268, pp. 268-179). In some embodiments, an immunostimulatory dose of an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, increases or is used to increase the antitumor activity of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule).

In some embodiments, an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule), are administered. at a dose and/or time schedule that, in combination, provides the desired antitumor activity.

In some embodiments, the dose of everolimus (Compound A36) approved for adult cancer indications (e.g., breast cancer (e.g., TNBC), renal cell carcinoma, and neuroendocrine tumors (e.g., atypical pulmonary carcinoid tumor) is 10 mg daily. Without limited by theory, in some embodiments, modeling and simulation based on mTOR-mediated phosphorylation of its downstream S6 kinase (S6K) is believed to indicate that a 20 mg once weekly regimen inhibits mTOR-mediated S6K phosphorylation almost completely, a regimen with administration of 5 mg once a week inhibits S6K phosphorylation by more than 50% and a regimen of 0.5 mg once a day inhibits S6K phosphorylation by approximately 38% between doses (Mannick et al. (2014) Sci. Transl. Med Vol.6, Issue 268, pp. 268ra179 Tanaka (2008) J. Clin. Oncol. p. 1596-602. Thus, in some an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 5 mg once a week (QW), for example in combination with a PD-1 inhibitor (for example, an anti-PD antibody molecule -1). In other embodiments, a dose of 0.5 mg once a day (QD) may also be used, for example, if a dose of 5 mg once a week (QW) is not well tolerated. In some embodiments, an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 5 mg or less once a week.

In one embodiment, a TOR inhibitor or everolimus (Compound A36) is administered at a dose of about 2.5-20 mg/day. In one embodiment, the compound is administered at a dose of about 2.5, 5, 10, or 20 mg/day, such as about 2.5-5, 5-10, or 10-20 mg/day.

In one embodiment, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 1 mg to 10 mg, 2 mg to 8 mg, 3 mg to 7 mg, or 4 mg to 6 mg, such as at a dose of 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, or 10 mg, such as once a week, once a two weeks, once every three weeks, or once every four weeks. In one embodiment, the mTOR inhibitor, everolimus (Compound A36), or the compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 4 mg to 6 mg, eg 5 mg, eg once a week.

In another embodiment, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 0.1 mg to 1 mg, 0.2 mg to 0.8 mg, 0 .3 mg to 0.7 mg or 0.4 mg to 0.6 mg, e.g. 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg or 1 mg, for example, once a day or once a week.

In some embodiments, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered orally.

In some embodiments, the mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered at a dose of 2 mg to 8 mg (e.g., 5 mg), e.g., once a week, e.g., orally, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg dose), e.g., once every 4 weeks, e.g., by intravenous infusion .

In some embodiments, an mTOR inhibitor, everolimus (Compound A36), or a compound disclosed in PCT publication WO 2014/085318, is administered in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) for the treatment of colon and rectal cancer. (eg, MSS CRC), lung cancer (eg, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple negative breast cancer (TNBC)).

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an inhibitor of one or more of VEGFR-2, PDGFR beta, KIT, or RAF kinase C, 1 -methyl-5-((2-(5-(trifluoromethyl)-1H-imidazol-2-yl)pyridin-4-yl)oxy)-N-(4-(trifluoromethyl)phenyl)-1H-benzo[d] imidazol-2-amine (Compound A37), or a compound disclosed in PCT publication WO 2007/030377, for the treatment of a disorder, for example, the disorder described herein. In one embodiment, the inhibitor of one or more of VEGFR-2, PDGFR beta, KIT, or RAF kinase C is 1-methyl-5-((2-(5-(trifluoromethyl)-1H-imidazol-2-yl)pyridine- 4-yl)oxy)-N-(4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazol-2-amine (Compound A37) or the compound disclosed in PCT Publication WO 2007/030377. In one embodiment, the PD-1 antibody molecule is used in combination with 1-methyl-5-((2-(5-(trifluoromethyl)-1H-imidazol-2-yl)pyridin-4-yl)oxy)-N-( 4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazol-2-amine (Compound A37) or a compound disclosed in PCT publication WO 2007/030377 for the treatment of a disorder such as cancer, melanoma or solid tumor.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an inhibitor of somatostatin and/or growth hormone release agonists, pasireotide diaspartate (also known as Signifor; Compound A38 ), or a compound disclosed in PCT Publication WO2002/010192 or US Pat. No. 7,473,761, for the treatment of a disorder, for example, the disorder described herein. In one embodiment, the inhibitor of somatostatin agonists and/or growth hormone release is pasireotide diaspartate (Compound A38) or a compound disclosed in PCT Publication WO2002/010192 or US Patent 7,473,761. In one embodiment, a PD-1 antibody molecule is used in combination with pasireotide diaspartate (Compound A38) or a compound disclosed in PCT Publication WO2002/010192 or US Pat. No. 7,473,761 to treat a disorder such as prostate cancer, endocrine cancer, neurological cancer, neuroendocrine tumor (NET) (eg, atypical pulmonary carcinoid tumour), skin cancer (eg, melanoma or Merkel cell cancer), pancreatic cancer, liver cancer, Cushing's syndrome, gastrointestinal disorder, acromegaly, liver and bile duct disease, or cirrhosis of the liver.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a signal transduction modulator and/or angiogenesis inhibitor, dovitinib (Compound A39), or a compound disclosed in PCT publication WO 2009/115562, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the signal transduction modulator and/or angiogenesis inhibitor is dovitinib (Compound A39) or a compound disclosed in PCT publication WO 2009/115562. In one embodiment, a PD-1 antibody molecule is used in combination with dovitinib (Compound A39) or a compound disclosed in PCT publication WO 2009/115562 to treat a disorder such as cancer, respiratory/breast cancer, multiple myeloma, prostate cancer , non-small cell lung cancer, endocrine cancer, or a neurological genetic disorder.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an EGFR inhibitor, (R,E)-N-(7-chloro-1-(1 -(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013 /184757, for the treatment of disorders, for example, disorders described in this application. In one embodiment, the EGFR inhibitor is (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40) or the compound disclosed in PCT publication WO 2013/184757. In one embodiment, the PD-1 antibody molecule is used in combination with (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)- 1H-benzo[d]imidazol-2-yl)-2-methylisonicotin-amide (Compound A40) or a compound disclosed in PCT publication WO 2013/184757 for the treatment of a disorder such as cancer, for example a solid tumor.

In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is a covalent, irreversible tyrosine kinase inhibitor. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, inhibits EGFR activating mutations (L858R, ex19del). In other embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, does not or substantially does not inhibit wild-type (wt) EGFR. Compound A40 has shown efficacy in patients with EGFR mutant NSCLC. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, also inhibits one or more kinases from the TEC kinase family. The Tec family kinases include, for example, ITK, BMX, TEC, RLK, and BTK, and are central to T-cell receptor proliferation and chemokine receptor signaling (Schwartzberg et al. (2005) Nat. Rev. Immunol. p. 284-95) . For example, Compound A40 can inhibit ITK with a biochemical IC _{50 of} 1.3 nM. ITK is an essential enzyme for the survival of Th2 cells, and its inhibition results in a shift in the balance between Th2 and Th1 cells. Combination in vivo treatment with the ITK inhibitor ibrutinib or Compound A40 and anti-PD-L1 antibody results in superior efficacy compared to either agent alone in several models.

In some embodiments, the combination described herein includes an EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepane-3-yl )-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g. , an anti-PD-1 antibody molecule).

The combination of ITK inhibition (ibrutinib) and checkpoint inhibition is more effective than either agent alone in various syngeneic mouse models, such as those expressing ITK but not expressing BTK. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, inhibits ITK. The synergistic effect of ITK inhibition and checkpoint blockade was investigated in mouse allografts using mouse cancer cell lines (A20, CT26 and 4T1) (Sagiv-Barfi et al. (2015) Blood, pp. 2079-86). The combination of an anti-PD-L1 antibody and ibrutinib (an ITK inhibitor) was shown to be significantly more effective than either agent alone in all three models. In these experiments, the treatment effect was sustained despite administration of ibrutinib for only 8 days and administration of a total of 5 doses of anti-PD-L1 antibody. Approximately half of the mice with CT26 tumors treated with this combination were completely cured (none of the mice treated with any single agent were cured). Repeated administration of CT26 tumor inoculum to these mice demonstrated long-term antitumor memory specific to this cell line (Sagiv-Barfi et al. (2015) Blood, pp. 2079-86). In addition, tumor-specific T cells were detected in the blood and spleen of mice treated with ibrutinib and anti-PD-L1 antibody. To further explore this observation for Compound A40, a similar experiment was performed using the A20 lymphoma model. In this study, the combination of Compound A40 and an anti-PD-L1 antibody or ibrutinib and an anti-PD-L1 antibody was more effective than either agent alone. Compound A40 and ibrutinib were administered for only ten days and a total of 5 doses of anti-PD-L1 antibody were administered. The effect of Compound A40 plus anti-PD-L1 antibody and ibrutinib plus anti-PD-L1 antibody on survival lasted longer than 60 days, even if Compound A40 and ibrutinib were administered only briefly. The combination of anti-PD-L1 antibody and Compound A40 resulted in tumor regression in A20 lymphoma allograft-bearing mice. Thus, in some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo [d]imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, enhances or is used to enhance the antitumor effect of a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule ).

In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT Publication WO 2013/184757, and an inhibitor of an immune checkpoint molecule, e.g., a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) , administered at a dose and/or time schedule that, in combination, provide the desired antitumor activity.

In one embodiment, an EGFR inhibitor or (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40) is administered at a dose of 150-250 mg, for example, per day. In one embodiment, the compound is administered at a dose of about 150, 200, or 250 mg, or about 150-200 or 200-250 mg.

In one embodiment, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40) or the compound disclosed in PCT publication WO 2013/184757 is administered at a dose of 5 mg to 100 mg, e.g. 10 mg to 75 mg, 15 mg to 50 mg, 20 mg to 30 mg, 10 mg to 40 mg, 10 mg to 25 mg, or 25 mg to 40 mg, e.g. at 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30mg, 35mg, 40mg, 45mg, 50mg, 60mg, 70mg, 80mg, 90mg or 100mg e.g. twice a day, once a day, once every other day, once three days or once a week.

In one embodiment, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40) or the compound disclosed in PCT publication WO 2013/184757 is administered at a dose of 10 mg to 50 mg (eg 25 mg) once a day. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is administered orally. In one embodiment, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT Publication WO 2013/184757, is administered at a dose of 10 mg to 50 mg (e.g. 25 mg), for example, once a day, e.g., orally, and a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) is administered at a dose of 300 mg to 500 mg (e.g., 400 mg), e.g., once every 4 weeks, e.g., by intravenous infusion . In some embodiments, the combination is administered in one or more courses of use, eg, one or more 28 day courses of use. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is administered on day 1 to day 10 of the first course of application. In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or the compound disclosed in PCT publication WO 2013/184757, is not administered on day 11 to day 28 of the first course of application, or is not administered in any of the subsequent courses of application.

In some embodiments, the EGFR inhibitor, (R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d] imidazol-2-yl)-2-methylisonicotinamide (Compound A40), or a compound disclosed in PCT publication WO 2013/184757, is administered in combination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule) for the treatment of colon cancer and rectal cancer (CRC), lung cancer (eg, non-small cell lung cancer (NSCLC)) or breast cancer (eg, triple-negative breast cancer (TNBC)).

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an ALK inhibitor, N ⁶ -(2-isopropoxy-5-methyl-4-(1- methylpiperidin-4-yl)phenyl)-N ⁴ -(2-(isopropylsulfonyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidin-4,6-diamine (Compound A42), or a compound disclosed in PCT Publication WO 2008/073687, for the treatment of a disorder, such as the disorder described herein. In one embodiment, the ALK inhibitor is N ⁶ -(2-isopropoxy-5-methyl-4-(1-methylpiperidin-4-yl)phenyl)-N ⁴ -(2-(isopropylsulfonyl)phenyl)-1H-pyrazolo[3 ,4-d]pyrimidine-4,6-diamine (Compound A42) or the compound disclosed in PCT Publication WO 2008/073687. In one embodiment, the PD-1 antibody molecule is used in combination with N ⁶ -(2-isopropoxy-5-methyl-4-(1-methylpiperidin-4-yl)phenyl)-N ⁴ -(2-(isopropylsulfonyl)-phenyl )-1H-pyrazolo[3,4-d]pyrimidine-4,6-diamine (Compound A42) or a compound disclosed in PCT Publication WO 2008/073687 for the treatment of a disorder such as cancer, anaplastic large cell lymphoma (ALCL), non-small cell lung carcinoma (NSCLC) or neuroblastoma.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an IGF-1R inhibitor, 3-(4-(4-((5-chloro-4 -((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)amino)-5-fluoro-2-methylphenyl)piperidin-1-yl)thietan-1,1-dioxide (Compound A43 ), 5-chloro-N ² -(2-fluoro-5-methyl-4-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)phenyl)-N ⁴ -(5-methyl -1H-pyrazol-3-yl)pyrimidin-2,4-diamine (Compound A44) or 5-chloro-N2-(4-(1-ethylpiperidin-4-yl)-2-fluoro-5-methylphenyl)-N ^4- (5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (Compound A45), or a compound disclosed in PCT publication WO 2010/002655, for the treatment of a disorder, for example, the described disorder. In one embodiment, the IGF-1R inhibitor is 3-(4-(4-((5-chloro-4-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)amino)- 5-fluoro-2-methylphenyl)piperidin-1-yl)thietan-1,1-dioxide (Compound A43), 5-chloro-N ² -(2-fluoro-5-methyl-4-(1-(tetrahydro- 2H-pyran-4-yl)piperidin-4-yl)phenyl)-N ⁴ -(5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (Compound A44), 5-chloro-N2 -(4-(1-ethylpiperidin-4-yl)-2-fluoro-5-methylphenyl)-N ⁴ -(5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (Compound A45) or a compound disclosed in PCT publication WO 2010/002655. In one embodiment, the PD-1 antibody molecule is used in combination with 3-(4-(4-((5-chloro-4-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl )amino)-5-fluoro-2-methylphenyl)piperidin-1-yl)thietan-1,1-dioxide (Compound A43), 5-chloro-N ² -(2-fluoro-5-methyl-4-(1 -(tetrahydro-2H-pyran-4-yl)-piperidin-4-yl)phenyl)-N ⁴ -(5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4-diamine (Compound A44), 5-chloro-N2-(4-(1-ethylpiperidin-4-yl)-2-fluoro-5-methylphenyl)-N ⁴ -(5-methyl-1H-pyrazol-3-yl)pyrimidin-2,4- diamine (Compound A45) or a compound disclosed in PCT publication WO 2010/002655 for the treatment of a disorder such as cancer or sarcoma.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a P-glycoprotein 1 inhibitor, valspodar (also known as AMDRAY; Compound A46), or a compound disclosed in EP 296122, for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the P-glycoprotein 1 inhibitor is valspodar (Compound A46) or a compound disclosed in EP 296122. In one embodiment, a PD-1 antibody molecule is used in combination with valspodar (Compound A46) or a compound disclosed in EP 296122 to treating a disorder such as a drug-resistant cancer or tumor.

In another embodiment, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with one or more of a VEGFR inhibitor, vatalanib succinate (Compound A47), or a compound disclosed in EP 296122 , for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the VEGFR inhibitor is vatalanib succinate (Compound A47) or a compound disclosed in EP 296122. In one embodiment, a PD-1 antibody molecule is used in combination with vatalanib succinate (Compound A47) or a compound disclosed in EP 296122 cancer.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with an IDH inhibitor or a compound disclosed in WO2014/141104 for the treatment of a disorder, e.g., the disorder described in this application. In one embodiment, the IDH inhibitor is a compound disclosed in PCT Publication WO2014/141104. In one embodiment, a PD-1 antibody molecule is used in combination with a compound disclosed in WO2014/141104 to treat a disorder such as cancer.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a BCL-ABL inhibitor or a compound disclosed in PCT Publication WO2013/171639, WO2013/171640, WO2013 /171641 or WO2013/171642, for the treatment of a disorder, for example, the disorder described in this application. In one embodiment, the BCL-ABL inhibitor is a compound disclosed in PCT Publication WO2013/171639, WO2013/171640, WO2013/171641, or WO2013/171642. In one embodiment, a PD-1 antibody molecule is used in combination with a compound disclosed in PCT Publication WO2013/171639, WO2013/171640, WO2013/171641 or WO2013/171642 to treat a disorder such as cancer.

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a c-RAF inhibitor or a compound disclosed in PCT Publication WO2014/151616, to treat a disorder, e.g. , the violation described in this application. In one embodiment, the c-RAF inhibitor is Compound A50 or a compound disclosed in PCT Publication WO2014/151616. In some embodiments, the c-RAF inhibitor or Compound A50 is a compound of formula (I):

or a pharmaceutically acceptable salt thereof, where:

Z ¹ is O, S, S(=O) or SO ₂ ;

Z ² is N, S or CR ^a where R ^a is H, halogen, C _1-4 alkyl or C _1-4 haloalkyl;

R ¹ is CN, halogen, OH, C _1-4 alkoxy or C _1-4 alkyl which is optionally substituted with one to three groups selected from halogen, C _1-4 alkoxy, CN and hydroxyl;

Ring B is selected from phenyl, pyridine, pyrimidine, pyrazine, pyridazine, pyridone, pyrimidone, pyrazinone, pyridazinone and thiazole, each of which is optionally substituted with no more than two groups selected from halo, OH, CN, C _1-4 alkyl, C _2-4 alkenyl, -O-(C _1-4 alkyl), NH ₂ , NH-(C _1-4 alkyl), -N(C _1-4 alkyl) ₂ , -SO ₂ R ² , NHSO ₂ R ² , NHC(O)R ² , NHCO ₂ R ² , C _3-6 cycloalkyl, 5-6 membered heteroaryl, -OC _3-6 cycloalkyl, -O-(5-6 membered heteroaryl), C _4-8 heterocycloalkyl and -O-(4-8 membered heterocycloalkyl), where each heterocycloalkyl and heteroaryl contain up to three heteroatoms selected from N, O and S as ring members,

where each C _1-4 alkyl, C _2-4 alkenyl, C _3-6 cycloalkyl, 5-6 membered heteroaryl and 4-8 membered heterocycloalkyl are optionally substituted with no more than three groups selected from oxo, hydroxyl, halogen, C _1-4 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy and -(CH ₂ ) _1-2 Q, where Q is OH, C _1-4 alkoxy, -CN, NH ₂ , -NHR ³ , -N (R ³ ) ₂ , -SO ₂ R ³ , NHSO ₂ R ³ , NHC(O)OR ³ or NHC(O)R ³ ; each R ² and R ³ is independently C _1-4 alkyl; and

Ring B is optionally fused to a 5-6 membered aromatic or non-aromatic ring containing up to two heteroatoms selected from N, O and S, where the 5-6 membered ring may be substituted with halogen, C _1-4 alkyl, C _1-4 haloalkyl or C _1-4 alkoxy, and if the fused ring is non-aromatic, substituent variants may further include oxo;

each Y is independently selected from C _1-4 alkyl, C _1-4 alkoxy, CN, halogen, oxo, -(CH2) _p OR ⁴ , -(CH ₂ ) _p N(R ⁴ ) ₂ , -(CH ₂ ) _p NHC(O)R ⁴ , -(CH ₂ ) _p NHCOO(C _1-4 alkyl) and imidazole,

or two Y groups on Ring A optionally combined to form a ring fused to or bridged by Ring A, wherein said fused or bridged ring optionally contains a heteroatom selected from N, O and S as a ring member and is optionally substituted with no more than than two groups selected from C _1-4 alkyl, C _1-4 alkoxy, CN, halogen, oxo, -(CH ₂ ) _p OR ⁴ , -(CH ₂ )pN(R ⁴ ) ₂ , -(CH ₂ ) _p NHC(O)R ⁴ and -(CH ₂ ) _p NHCOO(C _1-4 alkyl);

each R ⁴ is independently H or C _1-4 alkyl;

each p is independently 0, 1, or 2;

q is 0, 1 or 2;

Z ³ , Z ⁴ and Z ⁵ are independently selected from CH and N and optionally NO;

L is -C(=O)-NR ⁴ -[CY] or -NR ⁴ -C(=O)-[CY], where [CY] indicates which L atom is attached to CY; and

CY is an aromatic ring selected from phenyl, pyridine, pyrimidine, pyrazine, pyridazine, pyridone, thiazole, isothiazole, oxazole, pyrazole and isoxazole, where the ring is optionally fused to a thiophene, imidazole, oxazolone or pyrrole ring;

and CY is substituted with no more than two groups selected from halo, CN, R ⁵ , OR ⁵ , SO ₂ R ⁵ , -S(=NH)(=O)R ⁵ , OH, NH ₂ , NHR ⁵ and -N( R ⁵ ) ₂ ,

where each R ⁵ is independently C _1-4 alkyl, C _2-4 alkenyl, C _2-6 heterocyclyl, 5-membered heteroaryl containing up to three heteroatoms selected from N, O and S as ring members, or C _{3 -8} cycloalkyl, and R ⁵ is optionally substituted with no more than four groups selected from oxo, halo, CN, R ⁶ , OH, OR ⁶ , SO ₂ R ⁶ , NH ₂ , NHR ⁶ , N(R ⁶ ) ₂ , NHSO ₂ R ⁶ , NHCOOR ⁶ , NHC(=O)R ⁶ , -CH ₂ OR ⁷ , -CH ₂ N(R ⁷ ) ₂ , where each

R ⁶ is independently C _1-4 alkyl and each R ⁷ is independently H or C _1-4 alkyl;

and two R ⁴ , R ⁵ , R ⁶ or R ⁷ on the same nitrogen atom may be combined to form a 5-6 membered heterocyclic ring, optionally containing an additional N, O or S as a ring member, and optionally substituted with no more than two groups selected from C _1-4 alkyl, oxo, halogen, OH and C _1-4 alkoxy.

In one embodiment, a PD-1 antibody molecule is used in combination with a compound disclosed in PCT Publication WO2014/151616 to treat a disorder such as cancer, such as the cancer described herein. In some embodiments, the cancer is lung cancer (eg, non-small cell lung cancer).

In another embodiment, a combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, comprises or is used in combination with a competitive inhibitor of ERK1/2 ATP, or a compound disclosed in PCT Publication WO2015/066188, for the treatment violations, for example, the violations described in this application. In one embodiment, the competitive ERK1/2 ATP inhibitor is a compound disclosed in PCT Publication WO2015/066188. In one embodiment, a PD-1 antibody molecule is used in combination with Compound A51 or a compound disclosed in PCT publication WO2015/066188 to treat a disorder such as cancer.

In some embodiments, the combination, for example, a combination comprising an anti-PD-1 antibody molecule as described herein, is administered in combination with one or more of Compound A8, Compound A17, Compound A23, Compound A24, Compound A27, Connections A29 and Connections A33.

In some embodiments, the combination, e.g., a combination comprising an anti-PD-1 antibody molecule as described herein, is administered in combination with an antineoplastic agent having known activity in an immune cell assay, e.g., in one or more of the huMLR assay, T cell proliferation assay; and B cell proliferation assay. Examples of assays are described below. Based on the analysis, an IC50 value can be calculated for each test agent. In embodiments, the anticancer agent has an IC50, eg, 0-1 µM, 1-4 µM, or greater than 4 µM, eg, 4-10 µM or 4-20 µM. In embodiments, 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.

In some embodiments, Compound A28 (or a related compound to Compound A28) is administered at a dose of about 5-10 or 10-30 mg. In some embodiments, Compound A22 (or a related compound to Compound A22) is administered at a dose of approximately 200 mg. In some embodiments, Compound A17 (or a related compound to Compound A17) is administered at a dose of about 400-600 mg. In some embodiments, Compound A16 (or a related compound to Compound A16) is administered orally at a dose of about 400-600 mg per day. In some embodiments, Compound A29 (or a related compound to Compound A29) is administered at a dose of about 200-400 or 300-400 mg. In some embodiments, Compound A24 (or a related compound to Compound A24) is administered at a dose of about 200-600 mg. In some embodiments, Compound A23 (ceritinib) (or a ceritinib-related compound) is administered at a dose of approximately 750 mg once daily. In some embodiments, Compound A8 (or a related compound to Compound A8) is administered at a dose of about 200-400 or 300-400 mg. In some embodiments, Compound A5 (or a related compound to Compound A5) is administered at a dose of about 100-125 mg. In some embodiments, Compound A6 (or a related compound to Compound A6) is administered at a dose of about 100 mg. In some embodiments, Compound A1 (or a related compound to Compound A1) is administered at a dose of about 200-300 or 200-600 mg. In some embodiments, Compound A40 (or a related compound to Compound A40) is administered at a dose of about 150-250 mg. In embodiments, Compound A10 (or a compound related to Compound A10) is administered at a dose of about 400-700 mg, eg, administered three times a week for 2 weeks and one week apart. In embodiments, the BCR-ABL inhibitor is administered at a dose of about 20 mg twice daily - 80 mg twice daily.

Examples of huMLR assays and B or T cell proliferation assays are provided below.

Mixed culture reaction of human lymphocytes

The mixed lymphocyte culture test (MLC) is a functional test that measures the proliferative response of lymphocytes from one individual (responder) to lymphocytes from another individual (stimulator). To perform allogeneic SCL, peripheral blood mononuclear cells (PBMCs) from three donors were isolated from a buffy coat with HLA of unknown type (Kantonspital Blutspendezentrum from Bern and Aarau, Switzerland). Cells were prepared at a concentration of 2×10 ⁵ in 0.2 ml culture medium containing RPMI 1640 GlutaMAX ^TM with 10% fetal calf serum (FCS), 100 U penicillin/100 μg streptomycin, 50 μM 2-mercaptoethanol. Bidirectional SCL donor reactions were performed by mixing PBMCs from two different donors at a 1:1 ratio and cocultivating in triplicate in flat-bottomed 96-well tissue culture plates for 6 days at 37°C, 5% CO ₂ , in the presence of test compounds in range of 8 concentrations or none. Cells were pulsed with 3H-TdR (1 μCi/0.2 ml) during the last 16 hours of culture and radiolabeling was used to measure cell proliferation. The concentration at which 50% inhibition of the maximum huMLR response (IC50) occurs was calculated for each compound. Cyclosporine was used as a positive control for huMLR inhibition.

Human B cell proliferation assay

PBMCs were isolated in a Ficoll-Paque density gradient from fresh human blood and subjected to negative B-cell isolation. B cells were resuspended in culture medium (RPMI 1640, HEPES, 10% FCS, 50 µg/ml gentamicin, 50 µM 2-mercaptoethanol, 1×ITS (insulin, transferrin and sodium selenite), 1× non-essential amino acids) at a concentration of 9× 10 ⁴ per well in flat-bottomed 96-well culture plates. B cell stimulation was performed using human anti-IgM antibody molecule (30 µg/mL) and IL-4 (75 ng/mL) or CD40 ligand (3 µg/mL) and IL-4 (75 ng/mL) in the presence of test subjects. compounds in a range of 7 concentrations or none. After 72 hours of culture at 37° C., 10% CO ₂ , cells were pulsed with 3H-TdR (1 μCi/well) during the last 6 hours of culture. B cells were then harvested and thymidine incorporation was measured using a scintillation counter. Average values were calculated for each replication of treatments and the data were entered into the XLfit 4 program to determine the corresponding IC50 values.

Human T cell proliferation assay

PBMCs were isolated in a Ficoll-Paque density gradient from fresh human blood and subjected to negative T cell isolation. T cells were prepared in culture medium (RPMI 1640, HEPES, 10% FCS, 50 μg/ml gentamicin, 50 μM 2-mercaptoethanol, 1×ITS (insulin, transferrin and sodium selenite), 1× non-essential amino acids) at a concentration of 8× 10 ⁴ per well in flat-bottomed 96-well culture plates. T cell stimulation was performed using a human anti-CD3 antibody molecule (10 μg/ml) or a human anti-CD3 antibody molecule (5 μg/ml) and an anti-CD28 antibody molecule (1 μg/ml) in the presence of test compounds in a range of 7 concentrations or in their absence. After 72 hours of culture at 37° C., 10% CO ₂ , cells were pulsed with 3H-TdR (1 μCi/well) during the last 6 hours of culture. Cell proliferation was measured by thymidine incorporation, allowing an IC50 determination for each test compound.

Contraception

In some embodiments of any of the methods disclosed herein, the subject uses a contraceptive (e.g., a contraceptive) before, during, or after use of a therapy (e.g., combination therapy) comprising an anti-PD-1 antibody molecule described herein. . Without being limited by theory, it is believed that in some embodiments, the use of contraception for some period before use, during or after the use of therapy (for example, combination therapy) can reduce or prevent the risk of embryofetal toxicity, for example, an increased risk of miscarriage or death of a premature baby .

In some embodiments, the method described herein comprises administering a contraceptive to a subject (e.g., female) who is or has been using a therapy (e.g., combination therapy) comprising an anti-PD-1 antibody molecule described herein. In other embodiments, a contraceptive is used in combination with a therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule described herein for the treatment of a disorder described herein, such as a cancer described herein. In other embodiments, a contraceptive is used in combination with a therapy (e.g., combination therapy) comprising an anti-PD-1 antibody molecule described herein to reduce or prevent the risk of embryofetal toxicity, e.g., an increased risk of miscarriage or premature death.

In some embodiments, the subject is instructed, for example, by a healthcare provider, to discontinue therapy (e.g., combination therapy) comprising an anti-PD-1 antibody molecule if the subject is or is known to be pregnant, or there is a high likelihood that that the subject is pregnant. In other embodiments, the subject discontinues therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule if the subject is pregnant, or the subject is known to be pregnant, or there is a high probability that the subject is pregnant.

In some embodiments, the length of time that a subject (e.g., female) is using effective contraception (sometimes referred to herein as the contraceptive period) can be determined at least in part by the half-life (T ¹ / ₂ ) of the anti- Serum PD-1. During the period of contraception, the subject may complete therapy or continue therapy. In some embodiments, the period of contraception includes the period following the last dose of therapy (eg, the last dose of an anti-PD-1 antibody molecule). In other embodiments, the period of contraception further includes the period during which the subject receives therapy (eg, an anti-PD-1 antibody molecule).

In some embodiments, the contraceptive period includes a period of 50 days or more, such as 60 days or more, 80 days or more, 90 days or more, 100 days or more, 110 days or more, 120 days or more, 130 days or more , 140 days or more, 150 days or more, 160 days or more, 170 days or more, 180 days or more, 190 days or more, 200 days or more, 220 days or more, 240 days or more, 260 days or more , 280 days or more, or 300 days or more, for example, after the last dose of the anti-PD-1 antibody molecule. In some embodiments, the period of contraception includes a period of 150 days or more, such as after the last dose of the anti-PD-1 antibody molecule. In some embodiments, the period of contraception includes a period when the subject receives the anti-PD-1 antibody molecule and a period of 150 days or more after the last dose of the anti-PD-1 antibody molecule.

In some embodiments, the contraceptive period is at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 times the serum half-life of the anti-PD-1 antibody molecule. In some embodiments, the contraceptive period is at least 5 times the serum half-life of the anti-PD-1 antibody molecule. In some embodiments, the serum half-life of an anti-PD-1 antibody molecule is 10 days to 30 days, such as 15 days to 25 days, 17 days to 23 days, or 19 days to 21 days, such as 15 days or more, 16 days or more, 17 days or more, 18 days or more, 19 days or more, 20 days or more, 21 days or more, 22 days or more, or 23 days or more. In some embodiments, the serum half-life of an anti-PD-1 antibody molecule is 17 days to 23 days, such as about 19 days, 20 days, 21 days, 22 days, or 23 days.

In some embodiments, the subject is using contraception prior to receiving therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule. In other embodiments, the subject is using contraception during therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule. In other embodiments, the subject is using contraception after receiving a therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule.

In some embodiments, the subject is using contraception during and after receiving therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule. In other embodiments, the subject is using contraception prior to and while receiving therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule. In other embodiments, the subject is using contraception before and after receiving a therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule. In other embodiments, the subject uses contraception before, during, and after receiving therapy (eg, combination therapy) comprising an anti-PD-1 antibody molecule.

In some embodiments, the subject uses contraception at least 1, 3, 7, 14, or 28 days or at least 1, 2, or 3 months prior to administration of the anti-PD-1 antibody molecule to the subject, for example, prior to the administration of the first dose of the anti-PD-1 antibody molecule. PD-1.

In other embodiments, the subject uses contraception during therapy (e.g., combination therapy) at least one, two, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, or 10 times, during the period during which the anti-PD-1 antibody molecule is administered to the subject, for example between the first and last doses of the anti-PD-1 antibody molecule.

In other embodiments, the subject has been using contraception for at least 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, or 300 days, or at least 3, 4, 5, 6, 7, 9, or 10 months after administration of the anti-PD-1 antibody molecule to the subject, e.g., after the last dose of the anti-PD-1 antibody molecule.

The subject may use various types of contraception (eg, contraceptives). For example, contraception may be provided by a device, drug, method, or any combination thereof. Examples of types of contraception (e.g., contraceptives) that can be used by a subject include, but are not limited to, a contraceptive implant (e.g., etonogestrel), a contraceptive transdermal patch, an oral contraceptive (e.g., a progestin and/or estrogen), a contraceptive sponge (e.g. , medroxyprogesterone), contraceptive sponge, contraceptive vaginal ring, breastfeeding as birth control, cervical cap, condom, diaphragm, female condom, methods based on determining the period of fertility, intrauterine device (IUD or coil), postcoital oral contraceptive (emergency contraception ), extravaginal intercourse, spermicide, sterilization for women (sterilization by tubal ligation), vasectomy, coitus interruptus (a method of coitus interruption before ejaculation), or a combination.

In some embodiments, the implementation of the subject is a woman, for example, a woman with childbearing potential. A woman's ability to bear children may be influenced by various factors, such as age, health, genetics, pathological conditions or environmental factors. In some embodiments, the subject is between the ages of 15-50 years old, such as 18-45 years old, 20-40 years old, or 25-35 years old. In some embodiments, the subject is at least 20, 25, 30, 35, 40, or 45 years old.

Biomarkers

The invention also provides methods for evaluating or monitoring the effectiveness of a therapy (eg, combination therapy) described herein in a subject (eg, a subject having cancer, eg, the cancer described herein). The method includes obtaining an efficacy value for a therapy, where said value indicates the efficacy of the therapy.

In embodiments, the therapy effectiveness value includes one, two, three, four, five, six, seven, eight, nine, or more (e.g., all) of the following:

(i) a tumor infiltrating lymphocyte (TIL) phenotype parameter;

(ii) a myeloid cell population parameter;

(iii) a surface expression marker parameter;

(iv) an immunological response biomarker parameter;

(v) systemic cytokine modulation parameter;

(vi) free circulating DNA (scDNA) parameter;

(vii) systemic immunomodulation parameter;

(viii) microbiome parameter;

(ix) an activation marker parameter in a circulating immune cell; or

(x) circulating cytokine parameter.

In some embodiments, the TIL phenotype parameter includes the level or activity of one, two, three, four, or more (e.g., all) of the hematoxylin and eosin (HE) staining for TIL enumeration, on CD8, FOXP3, CD4, or CD3 in a subject, e.g., in a subject sample (eg, a tumor sample). In some embodiments, the myeloid cell population parameter includes the level or activity of one or both of CD68 or CD163 in a subject, eg, in a sample of the subject (eg, a tumor sample). In some embodiments, a surface expression marker parameter includes the level or activity of one, two, or more (e.g., all) of PD-L1, LAG-3, or TIM-3 in a subject, e.g., in a sample of the subject (e.g., a tumor sample). In some embodiments, the immunological response biomarker parameter includes the level or sequence of one or more nucleic acid-based markers in a subject, eg, in a sample of the subject (eg, a tumor sample). In some embodiments, the systemic cytokine modulation parameter includes the level or activity of one, two, three, four, five, six, seven, eight or more (e.g., all) of IL-18, IFN-γ, ITAC (CXCL11), IL- 6, IL-10, IL-4, IL-17, IL-15, or TGF-beta in a subject, eg, in a sample of the subject (eg, a blood sample, eg, a blood plasma sample). In some embodiments, the fsDNA parameter includes the sequence or level of one or more circulating tumor DNA (scDNA) molecules in a subject, e.g., in a sample of the subject (e.g., a blood sample, e.g., a blood plasma sample). In some embodiments, the systemic immunomodulation parameter includes a phenotypic analysis. an activated immune cell, eg, a CD3-expressing cell, a CD8-expressing cell, or both, in a subject, eg, in a sample of the subject (eg, eg, a blood sample, eg, a PBMC sample). In some embodiments, the microbiome parameter includes the sequence or expression level of one or more genes in the subject's microbiome, eg, in a sample of the subject (eg, a stool sample). In some embodiments, an activation marker parameter in a circulating immune cell includes the level or activity of one, two, three, four, five, or more (e.g., all) of circulating CD8+, HLA-DR+Ki67+, T cells, IFN-γ, IL -18 or CXCL11 (IFN-γ induced CCK) expressing cells in a sample (eg a blood sample, eg a blood plasma sample). In some embodiments, the circulating cytokine parameter includes the level or activity of IL-6 in the subject, eg, in a sample of the subject (eg, a blood sample, eg, a blood plasma sample).

In some embodiments, the evaluation or monitoring includes biomarker analysis. For example, the evaluation or monitoring of the effectiveness of therapy can be performed at the molecular level and/or cellular level for one or more of the biomarkers described in this application. In some embodiments, the marker change may be related to disease outcome. Examples of biomarkers and a set of samples are described in Table 8.

Table 8 . Biomarker Examples and Sample Set

sample type Visit/Point in time Volume Marker Target Recent tumor biopsy* Screening
(days -28 to -1) 3-6 needle biopsy punctures GE, CD8, FOXP3, CD4, CD3
CD68, CD163 expression
PD-L1, LAG-3, TIM-3
Based on nucleic acid (quantitative analysis or sequencing of DNA or RNA) Analysis of TIL phenotypes, myeloid cell population, surface expression markers
Assessment of other biomarkers of the immune response Course 3 Day 1-15
(Any time between day 1 and 15, with biopsy taken post-dose) with disease progression
(unscheduled)** Blood (plasma) Screening
(before dosing) 8 ml at time Systemic cytokines (eg, IL-18, IFN-γ, ITAC (CXCL11), IL-6, IL-10, IL-4, IL-17, IL-15, TGF-beta) Modulation of systemic cytokines C1D1 (after dosing) C1D15 (after dosing) C2D1 (after dosing) C2D15 (after dosing) Blood (plasma) Screening
(before dosing) 10 ml at time DNA sequencing in circulating tumor DNA (scDNA) Assessment of csDNA as a surrogate marker Completion of treatment
(Anytime) Blood (MCPC) Screening 8 ml at time Phenotypic analysis of activated immune cells (eg CD3, CD8) Systemic immunomodulation C1D1 after dosing C1D15 after dosing C2D1 after dosing C2D15 after dosing *If a recently acquired tumor specimen cannot be safely collected at enrollment, it may be replaced with an archival specimen if it has been acquired recently.
**Biopsy samples to assess progression of the disease are required to be collected at the time of progression, even if this was not done during the initial period of treatment.

In some embodiments, a test paired tumor sample is obtained to evaluate the pharmacodynamic effect on the tumor microenvironment. In some embodiments, tumor samples are obtained at screening and after approximately two courses of therapy. In other embodiments, a tumor sample is obtained at the time of disease progression, which occurs during treatment.

In some embodiments, the primary evaluation of a hematoxylin and eosin (HE) stained preparation for tumor infiltrating lymphocytes (TIL) is used to conduct a subsequent IHC analysis of the tumor microenvironment, e.g., IHC testing of one or more of the TIL lymphocytes (e.g., CD8, CD3 , CD4 or FOXP3), myeloid cell population (eg CD163 or CD68), a surface marker such as LAG-3, TIM-3 or PD-L1. Other techniques for assessing immunological response biomarkers, such as nucleic acid-based methods (sequencing or quantification of DNA or RNA) or other methods, can be applied in this rapidly developing field of science.

The extent of tumor infiltration by immune cells, including lymphocytes and macrophages, changes in regulatory T cell counts, and/or changes in gene expression in tumor biopsy specimens may determine the efficacy of a given combination.

In some embodiments, a blood sample is taken, as indicated in Table 8, to study the levels of circulating cytokines in blood plasma and an activation marker in circulating immune cells. Without being limited by theory, it is believed that in some embodiments, an increase in one or more of circulating CD8+, HLA-DR+Ki67+, T cells, IFN-γ, IL-18, or CXCL11 (IFN-γ induced CCK) expressing cells indicates expansion of a pre-existing primed immune response, while a decrease in IL-6 indicates reduced myeloid suppressor cells (MSCs) (Herbst et al. (2016) Lancet 387(10027):1540-50; Tumeh et al. (2014) Nature p. 568-71; Powles et al (2014) Nature p. 558-62). In some embodiments, a blood sample is obtained at screening and/or at the end of treatment for analysis of free circulating DNA (fsDNA), for example, to evaluate any correlation with disease outcome. Any additional markers or methods known in the art may also be used. In some embodiments, a combination of compound-specific markers is used to determine pharmacodynamic activity at lower doses used in therapy that are or may be lower than previously established doses.

Nucleic acids

The invention also provides nucleic acids comprising nucleotide sequences that encode heavy and light chain variable regions and CDR regions or hypervariable loops of anti-PD-1 antibody molecules as described herein. For example, the invention provides first and second nucleic acids encoding the heavy and light chain variable regions, respectively, of an anti-PD-1 antibody molecule selected from one or more of the antibody molecules disclosed herein. The nucleic acid may include the nucleotide sequence shown in the tables in this application, or a sequence substantially identical to it (for example, a sequence that is at least about 85%, 90%, 95%, 99% or more identical to it, or that differs not more than 3, 6, 15, 30 or 45 nucleotides from the sequences shown in the tables in this application.

In some embodiments, the nucleic acid may include a nucleotide sequence encoding at least one, two, or three CDR regions or hypervariable loops from a heavy chain variable region having the amino acid sequence shown in the tables herein, or a sequence substantially homologous thereto. (eg, a sequence that is at least about 85%, 90%, 95%, 99% or more identical to it, and/or having one or more substitutions, such as conservative substitutions). In other embodiments, the nucleic acid may include a nucleotide sequence encoding at least one, two, or three CDR regions or hypervariable loops from a light chain variable region having the amino acid sequence shown in the tables herein, or a sequence substantially homologous thereto. (eg, a sequence that is at least about 85%, 90%, 95%, 99% or more identical to it, and/or having one or more substitutions, such as conservative substitutions). In yet another embodiment, the nucleic acid may include a nucleotide sequence encoding at least one, two, three, four, five, or six CDR regions or hypervariable loops from the heavy and light chain variable regions having the amino acid sequence shown in the tables herein. application, or a sequence essentially homologous to it (eg, a sequence identical to it at least about 85%, 90%, 95%, 99% or more, and/or having one or more substitutions, such as conservative substitutions).

In some embodiments, the implementation of the nucleic acid may include a nucleotide sequence encoding at least one, two or three CDR regions or hypervariable loops from the variable region of a heavy chain having a nucleotide sequence shown in the tables in this application, a sequence essentially homologous to it ( for example, a sequence identical to it at least about 85%, 90%, 95%, 99% or more, and/or capable of hybridization under the stringency conditions described in this application). In another embodiment, the nucleic acid may include a nucleotide sequence encoding at least one, two, or three CDR regions or hypervariable loops from a light chain variable region having the nucleotide sequence shown in the tables herein, or a sequence substantially homologous thereto. (for example, a sequence identical to it at least about 85%, 90%, 95%, 99% or more, and/or capable of hybridization under the stringent conditions described in this application). In yet another embodiment, the nucleic acid may include a nucleotide sequence encoding at least one, two, three, four, five, or six CDR regions or hypervariable loops from the heavy and light chain variable regions having the nucleotide sequence shown in the tables herein. application, or a sequence essentially homologous to it (for example, a sequence identical to it at least about 85%, 90%, 95%, 99% or more, and / or capable of hybridization under the stringent conditions described in this application) .

In another aspect, the present application provides host cells and vectors containing the nucleic acids described in this application. Nucleic acids can be present in a single vector or separate vectors present in a single host cell or separate host cells, as described in more detail below.

In some embodiments, one or more nucleic acid molecules are provided that include one or both of the nucleotide sequences that encode heavy and light chain variable regions, CDR regions, hypervariable loops, and framework regions of anti-PD-1 antibody molecules. In some embodiments, the nucleotide sequence that encodes an anti-PD-1 antibody molecule is codon-optimized. For example, the invention provides first and second nucleic acids encoding the heavy and light chain variable regions, respectively, of an anti-PD-1 antibody molecule selected from one or more of, for example, any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04 BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 hum16, BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D, or BAP049-Clone-E as shown in Table 1, or a sequence substantially identical thereto. For example, the nucleic acid may include the nucleotide sequence shown in Tables 1 and 2, or a sequence substantially identical to it (e.g., a sequence that is at least about 85%, 90%, 95%, 99%, or more identical to it, or which differs by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Tables 1 and 2).

In other embodiments, the nucleic acid molecule comprises a nucleotide sequence that encodes a heavy chain variable domain and/or a heavy chain constant region comprising the amino acid sequence BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone- D or BAP049-Clone-E; or as described in Table 1; or the nucleotide sequence in Table 1; or a sequence that is substantially identical (eg, a sequence that is at least about 85%, 90%, 95%, 99% or more identical) to any of the above sequences.

In other embodiments, the nucleic acid molecule comprises a nucleotide sequence that encodes a light chain variable domain and/or a light chain constant region comprising the amino acid sequence BAP049-Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone- D or BAP049-Clone-E; or as described in Table 1; or the nucleotide sequence in Table 1; or a sequence that is substantially identical (eg, a sequence that is at least about 85%, 90%, 95%, 99% or more identical) to any of the above sequences.

The above nucleotide sequences encoding the heavy and light chain variable domain and the constant regions of an anti-PD-1 antibody may be present in a single nucleic acid molecule, or in a single nucleic acid molecule. In some embodiments, the implementation of the nucleic acid molecules include a nucleotide sequence encoding a leader sequence, for example, the leader sequence shown in Table 4, or a sequence essentially identical to it.

In some embodiments, the nucleic acid molecule comprises a nucleotide sequence encoding at least one, two, or three CDR regions or hypervariable loops from a heavy chain variable region having the amino acid sequence shown 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 to it, and/or having one, two, three, or more substitutions, insertions, or deletions, e.g., conservative substitutions).

In another embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding at least one, two, or three CDR regions or hypervariable loops from a light chain variable region having the amino acid sequence shown 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 to it, and/or having one, two, three, or more substitutions, insertions, or deletions, e.g., conservative substitutions).

In yet another embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding at least one, two, three, four, five, or six CDR regions or hypervariable loops from the heavy and light chain variable regions having the amino acid sequence shown in Table 1, or a sequence substantially homologous to it (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical to it, and/or having one, two, three or more substitutions, insertions or deletions, for example, conservative substitutions).

In one embodiment, the nucleic acid molecule comprises a nucleotide sequence encoding an anti-PD-1 antibody molecule that comprises a substitution in the light chain CDR3 at position 102 of the light chain variable region, e.g. position 102 of the light chain variable region according to Table 1 (e.g., SEQ ID NO: 16 or 24 for murine or chimeric, unmodified; or any of SEQ ID NO: 34, 42, 46, 54, 58, 62, 66, 70, 74 or 78 in the case of a modified sequence).

In another embodiment, the nucleic acid molecule comprises one or more heavy chain framework regions (e.g., any of VHFW1 (type a), VHFW1 (type b), VHFW2 (type a), VHFW2 (type b), VHFW2 (type c), VHFW3 (type a), VHFW3 (type b) or VHFW4, or any combination thereof, such as a combination of framework regions as described herein) for any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05 BAP049-hum06 BAP049-hum07 BAP049-hum08 BAP049-hum09 BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 Clone-A, BAP049-Clone-B, BAP049-Clone-C, BAP049-Clone-D or BAP049-Clone-E as shown in Tables 1 and 2, or a sequence substantially identical thereto. For example, a nucleic acid molecule may include the nucleotide sequence shown in Tables 1 and 2, or a sequence substantially identical to it (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical to it, or which differs by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Tables 1 and 2).

In another embodiment, the nucleic acid molecule comprises one or more light chain framework regions (e.g., any of VLFW1 (type a), VLFW1 (type b), VLFW1 (type c), VLFW1 (type d), VLFW1 (type e), VLFW2 (type a), VLFW2 (type b), VLFW2 (type c), VLFW3 (type a), VLFW3 (type b), VLFW3 (type c), VLFW3 (type d) or VLFW4, or any combination of these, e.g. , a combination of framework regions as described herein) for any of BAP049-hum01, BAP049-hum02, BAP049-hum03, BAP049-hum04, BAP049-hum05, BAP049-hum06, BAP049-hum07, BAP049-hum08, BAP049-hum09, BAP049-hum10 BAP049-hum11 BAP049-hum12 BAP049-hum13 BAP049-hum14 BAP049-hum15 BAP049-hum16 BAP049-Clone-A BAP049-Clone-B BAP049-Clone-C BAP049-Clone- D or BAP049-Clone-E as shown in Tables 1 and 2, or a sequence substantially identical thereto. For example, a nucleic acid molecule may include the nucleotide sequence shown in Tables 1 and 2, or a sequence substantially identical to it (e.g., a sequence that is at least about 85%, 90%, 95%, 99% or more identical to it, or which differs by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Tables 1 and 2).

In another embodiment, the nucleic acid molecule includes one or more heavy chain framework regions and one or more light chain framework regions, as described herein. The heavy and light chain framework regions may be present in the same vector or in separate vectors.

Vectors and host cells

In another aspect of the present application provides host cells and vectors containing the nucleic acids described in this application. Nucleic acids may be present in a single vector or separate vectors present in a single host cell or separate host cells.

In one embodiment, the vectors include nucleotides encoding the antibody molecule described in this application. In one embodiment, the vectors include the nucleotide sequences described in this application. Vectors include, but are not limited to, a virus, plasmid, cosmid, lambda phage, or yeast artificial chromosome (YAC).

Various vector systems may be used. For example, one class of vectors uses DNA elements that are derived from animal viruses such as, for example, bovine papilloma virus, polyoma virus, adenovirus, vaccinia virus, baculovirus, retroviruses (Rous sarcoma virus, MMTV or MOMLV) or SV40 virus. . Another class of vectors uses RNA elements derived from RNA viruses such as Semliki Forest virus, Eastern Equine Encephalitis virus and Flaviviruses.

In addition, cells that carry stably integrated DNA in their chromosomes can be selected by introducing one or more markers that allow selection of transfected host cells. The marker may confer, for example, prototrophy of the auxotrophic host, resistance to a biocide (eg, antibiotics), or resistance to heavy metals such as copper, and the like. The selectable marker gene can be directly linked to expressed DNA sequences or introduced into such a cell by co-transformation. Additional elements may also be required for optimal mRNA synthesis. These elements may include splicing signals as well as transcription promoters, enhancers, and termination signals.

Once the expression vector or DNA sequence containing the constructs has been prepared for expression, the expression vectors can be transfected or introduced into an appropriate host cell. Various techniques can be used to accomplish this, such as, for example, protoplast fusion, calcium phosphate precipitation, electroporation, retroviral transduction, viral transfection, gene gun, lipid-based transfection, or other standard techniques. In the case of protoplast fusion, the cells are grown in media and screened for the desired activity. Methods and conditions for culturing the resulting transfected cells and isolating the produced antibody molecule are known to those skilled in the art and may be modified or optimized depending on the particular expression vector and mammalian host cell used, based on the present disclosure.

The invention also provides host cells comprising a nucleic acid encoding an antibody molecule as described herein.

In one embodiment, the host cells are genetically modified to include nucleic acids encoding an antibody molecule.

In one embodiment, the host cells are genetically modified using an expression cassette. The phrase "expression cassette" refers to nucleotide sequences that are capable of inducing gene expression in host organisms that are compatible with such sequences. Such cassettes may include a promoter, an open reading frame with or without introns, and a termination signal. Additional factors necessary or useful in effecting expression may also be used, such as, for example, an inducible promoter.

The invention also provides host cells, including the vectors described in this application.

The cell may be, without limitation, a eukaryotic cell, a bacterial cell, an insect cell, or a human cell. Suitable eukaryotic cells include, but are not limited to, Vero cells, HeLa cells, COS cells, CHO cells, HEK293 cells, BHK cells, and MDCKII cells. Suitable insect cells include, but are not limited to, Sf9 cells.

In some embodiments, the host cell is 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 may be a cultured cell or cell line. Examples of mammalian cells include lymphocytic cell lines (eg, NSO), Chinese hamster ovary (CHO) cells, COS cells, oocytes, and transgenic animal cells, eg, mammary epithelial cells.

Table 1 . Amino acid and nucleotide sequences of mouse, chimeric and humanized antibody molecules. Antibody molecules include mouse mAb BAP049, chimeric mAbs BAP049-chi and BAP049-chi-Y, and humanized mAbs BAP049-hum01 to BAP049-hum16 and BAP049-Clone-A to BAP049-Clone-E. Shown are the amino acid and nucleotide sequences of the heavy and light chain CDR regions, the heavy and light chain variable regions, and the heavy and light chains.

BAP049HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 6 vh QVQLQQPGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTLVTVSA SEQ ID NO: 7 DNA VH CAGGTCCAGCTGCAGCAACCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA SEQ ID NO: 8 vh QVQLQQSGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTLVTVSA SEQ ID NO: 9 DNA VH CAGGTCCAGCTGCAGCAGTCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA BAP049LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 12 (Kebat) LCDR3 QNDYSYPCT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 15 (Chothia) LCDR3 DYSYPC SEQ ID NO: 16 VL DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLDSGNQKNFLTWYQQKPGQPPKLLIFWASTRESGVPDRFTGSGSVTDFTLTISSVQAEDLAVYYCQNDYSYPCTFGGGTKLEIK SEQ ID NO: 17 DNA VL GACATTGTGATGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGTAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGATTATAGTTATCCGTGCACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA BAP049-chi HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 18 vh QVQLQQPGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 19 DNA VH CAGGTCCAGCTGCAGCAGCCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 20 HC QVQLQQPGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 21 HC DNA CAGGTCCAGCTGCAGCAGCCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA SEQ ID NO: 22 vh QVQLQQSGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 23 DNA VH CAGGTCCAGCTGCAGCAGTCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 30 HC QVQLQQSGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 31 HC DNA CAGGTCCAGCTGCAGCAGTCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-chi LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 12 (Kebat) LCDR3 QNDYSYPCT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 15 (Chothia) LCDR3 DYSYPC SEQ ID NO: 24 VL DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLDSGNQKNFLTWYQQKPGQPPKLLIFWASTRESGVPDRFTGSGSVTDFTLTISSVQAEDLAVYYCQNDYSYPCTFGQGTKVEIK SEQ ID NO: 25 DNA VL GACATTGTGATGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGTAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGATTATAGTTATCCGTGCACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 26 LC DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLDSGNQKNFLTWYQQKPGQPPKLLIFWASTRESGVPDRFTGSGSVTDFTLTISSVQAEDLAVYYCQNDYSYPCTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSKDSTYSLSSTLTLSKADYEKHKVYACE SEQ ID NO: 27 DNA LC GACATTGTGATGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGTAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGATTATAGTTATCCGTGCACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-chi-Y HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 18 vh QVQLQQPGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 19 DNA VH CAGGTCCAGCTGCAGCAGCCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 20 HC QVQLQQPGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 21 HC DNA CAGGTCCAGCTGCAGCAGCCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA SEQ ID NO: 22 vh QVQLQQSGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 23 DNA VH CAGGTCCAGCTGCAGCAGTCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 30 HC QVQLQQSGSELVRPGASVKLSCKASGYTFTTYWMHWVRQRPGQGLEWIGNIYPGTGGSNFDEKFKNRTSLTVDTSSTTAYMHLASLTSEDSAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 31 HC DNA CAGGTCCAGCTGCAGCAGTCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCGTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGAGGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAAAACAGGACCTCACTGACTGTAGACACATCCTCCACCACAGCCTACATGCACCTCGCCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-chi-Y LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 34 VL DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLDSGNQKNFLTWYQQKPGQPPKLLIFWASTRESGVPDRFTGSGSVTDFTLTISSVQAEDLAVYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 35 DNA VL GACATTGTGATGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGTAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 36 LC DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLDSGNQKNFLTWYQQKPGQPPKLLIFWASTRESGVPDRFTGSGSVTDFTLTISSVQAEDLAVYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYTKHKPVACE SEQ ID NO: 37 DNA LC GACATTGTGATGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGTAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum01HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum01LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 42 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 43 DNA VL GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 44 LC EIVLTQSPATLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSKDSTYSLSSTLTLSKADYEKHKVYACE SEQ ID NO: 45 DNA LC GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum02HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum02LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 46 VL DIQMTQSPSSLSASVGDRVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 47 DNA VL GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGATCCCACCTCGATTCAGTGGCAGCGGGTATGGAACAGATTTTACCCTCACAATTAATAACATAGAATCTGAGGATGCTGCATATTACTTCTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 48 LC DIQMTQSPSSLSASVGDRVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYTKHKPVACE SEQ ID NO: 49 DNA LC GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGATCCCACCTCGATTCAGTGGCAGCGGGTATGGAACAGATTTTACCCTCACAATTAATAACATAGAATCTGAGGATGCTGCATATTACTTCTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum03HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 50 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 51 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 52 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 53 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum03LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 46 VL DIQMTQSPSSLSASVGDRVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 47 DNA VL GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGATCCCACCTCGATTCAGTGGCAGCGGGTATGGAACAGATTTTACCCTCACAATTAATAACATAGAATCTGAGGATGCTGCATATTACTTCTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 48 LC DIQMTQSPSSLSASVGDRVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGIPPRFSGSGYGTDFTLTINNIESEDAAYYFCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYTKHKPVACE SEQ ID NO: 49 DNA LC GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGATCCCACCTCGATTCAGTGGCAGCGGGTATGGAACAGATTTTACCCTCACAATTAATAACATAGAATCTGAGGATGCTGCATATTACTTCTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum04HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 50 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 51 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 52 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 53 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum04LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 54 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 55 DNA VL GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 56 LC EIVLTQSPATLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYTKHKNSFACE SEQ ID NO: 57 DNA LC GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum05HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum05LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 54 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 55 DNA VL GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 56 LC EIVLTQSPATLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYTKHKNSFACE SEQ ID NO: 57 DNA LC GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum06HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum06LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 58 VL DIVMTQTPLSLPVTPGEPASISCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 59 DNA VL GATATTGTGATGACCCAGACTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 60 LC DIVMTQTPLSLPVTPGEPASISCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKSSVY SEQ ID NO: 61 DNA LC GATATTGTGATGACCCAGACTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum07HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum07LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 62 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 63 DNA VL GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 64 LC EIVLTQSPATLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE SEQ ID NO: 65 DNA LC GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum08HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 50 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 51 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 52 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 53 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum08LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 66 VL EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 67 DNA VL GAAATTGTGCTGACTCAGTCTCCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 68 LC EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQECKDSTYSLSSTLTLSKADYTKHKNSFACE SEQ ID NO: 69 DNA LC GAAATTGTGCTGACTCAGTCTCCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum09HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum09LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 66 VL EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 67 DNA VL GAAATTGTGCTGACTCAGTCTCCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 68 LC EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQECKDSTYSLSSTLTLSKADYTKHKNSFACE SEQ ID NO: 69 DNA LC GAAATTGTGCTGACTCAGTCTCCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum10HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 50 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 51 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 52 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 53 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum10LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 70 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 71 DNA VL GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 72 LC EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKTHNKSSVY SEQ ID NO: 73 DNA LC GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum11HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum11LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 70 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 71 DNA VL GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 72 LC EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKTHNKSSVY SEQ ID NO: 73 DNA LC GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum12HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum12LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 74 VL DIQMTQSPSSLSASVGDRVTITCKSSQSLLDSGNQKNFLTWYLQKPGQSPQLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 75 DNA VL GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 76 LC DIQMTQSPSSLSASVGDRVTITCKSSQSLLDSGNQKNFLTWYLQKPGQSPQLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQRGDSKDSTYSLSSTLTLECSSTYSLSSTLTLECSSADYSFEKHKVY SEQ ID NO: 77 DNA LC GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum13HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 39 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 40 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 41 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum13LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 78 VL DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 79 DNA VL GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCCATCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTAACCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 80 LC DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQRGDSKDSTYSLSSTLVSKADYEKHKK SEQ ID NO: 81 DNA LC GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCCATCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTAACCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum14HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 82 vh QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 83 DNA VH CAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTACTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 84 HC QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 85 HC DNA CAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTACTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum14LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 70 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 71 DNA VL GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 72 LC EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKTHNKSSVY SEQ ID NO: 73 DNA LC GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum15HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 82 vh QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 83 DNA VH CAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTACTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 84 HC QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 85 HC DNA CAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCTGGCTACACATTCACCACTTACTGGATGCACTGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTACTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum15LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 66 VL EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 67 DNA VL GAAATTGTGCTGACTCAGTCTCCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 68 LC EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQECKDSTYSLSSTLTLSKADYTKHKNSFACE SEQ ID NO: 69 DNA LC GAAATTGTGCTGACTCAGTCTCCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-hum16HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 86 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQAPGQGLEWMGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 87 DNA VH GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCC SEQ ID NO: 88 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQAPGQGLEWMGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: 89 HC DNA GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCTGGCTACACATTCACCACTTACTGGATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGTAATATTTATCCTGGTACTGGTGGTTCTAACTTCGATGAGAAGTTCAAGAACAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGATGGACTACTGGGACGGGAGCTTATTGGGGCCAGGGCACCACCGTGACCGTGTCCTCCGCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCGTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCT CCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA BAP049-hum16LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 66 VL EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 67 DNA VL GAAATTGTGCTGACTCAGTCTCCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 68 LC EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQECKDSTYSLSSTLTLSKADYTKHKNSFACE SEQ ID NO: 69 DNA LC GAAATTGTGCTGACTCAGTCTCCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGCAAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATTGGGCATCCACTAGGGAATCTGGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGTCAGAATGATTATAGTTATCCGTACACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT BAP049-Clone-A HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 90 DNA VH GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGAGTCCCTGCGGATCTCCTGCAAGGGCTCTGGCTACACCTTCACCACCTACTGGATGCACTGGGTGCGACAGGCTACCGGCCAGGGCCTGGAATGGATGGGCAACATCTATCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAACAGAGTGACCATCACCGCCGACAAGTCCACCTCCACCGCCTACATGGAACTGTCCTCCCTGAGATCCGAGGACACCGCCGTGTACTACTGCACCCGGTGGACAACCGGCACAGGCGCTTATTGGGGCCAGGGCACCACAGTGACCGTGTCCTCT SEQ ID NO: 91 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 92 HC DNA GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGAGTCCCTGCGGATCTCCTGCAAGGGCTCTGGCTACACCTTCACCACCTACTGGATGCACTGGGTGCGACAGGCTACCGGCCAGGGCCTGGAATGGATGGGCAACATCTATCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAACAGAGTGACCATCACCGCCGACAAGTCCACCTCCACCGCCTACATGGAACTGTCCTCCCTGAGATCCGAGGACACCGCCGTGTACTACTGCACCCGGTGGACAACCGGCACAGGCGCTTATTGGGGCCAGGGCACCACAGTGACCGTGTCCTCTGCTTCTACCAAGGGGCCCAGCGTGTTCCCCCTGGCCCCCTGCTCCAGAAGCACCAGCGAGAGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCAAGACCTACACCTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGAGCAAGTACGGCCCACCCTGCCCCCCCTGCCCAGCCCCCGAGTTCCTGGGCGGACCCAGCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGACGTGTCCCAGGAGGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTTTAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGTAAGGTCTCCAACAAGGGCCTGCCAAGCAGCATCGAAAAGACCATCA GCAAGGCCAAGGGCCAGCCTAGAGAGCCCCAGGTCTACACCCTGCCACCCAGCCAAGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCAAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAGGCTGACCGTGGACAAGTCCAGATGGCAGGAGGGCAACGTCTTTAGCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCTGGGC BAP049-Clone-A LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 42 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 93 DNA VL GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCACTGTCTCCAGGCGAGAGAGCTACCCTGTCCTGCAAGTCCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCTACTGGGCCTCCACCCGGGAATCTGGCGTGCCCTCTAGATTCTCCGGCTCCGGCTCTGGCACCGAGTTTACCCTGACCATCTCCAGCCTGCAGCCCGACGACTTCGCCACCTACTACTGCCAGAACGACTACTCCTACCCCTACACCTTCGGCCAGGGCACCAAGGTGGAAATCAAG SEQ ID NO: 44 LC EIVLTQSPATLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSKDSTYSLSSTLTLSKADYEKHKVYACE SEQ ID NO: 94 DNA LC GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCACTGTCTCCAGGCGAGAGAGCTACCCTGTCCTGCAAGTCCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCTACTGGGCCTCCACCCGGGAATCTGGCGTGCCCTCTAGATTCTCCGGCTCCGGCTCTGGCACCGAGTTTACCCTGACCATCTCCAGCCTGCAGCCCGACGACTTCGCCACCTACTACTGCCAGAACGACTACTCCTACCCCTACACCTTCGGCCAGGGCACCAAGGTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCAAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCAGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC BAP049-Clone-B HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 95 DNA VH GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCAGGCTACACCTTCACTACCTACTGGATGCACTGGGTCCGCCAGGCTACCGGTCAAGGCCTCGAGTGGATGGGTAATATCTACCCCGGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAATAGAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCACTAGGTGGACTACCGGCACAGGCGCCTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: 91 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 96 HC DNA GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCAGGCTACACCTTCACTACCTACTGGATGCACTGGGTCCGCCAGGCTACCGGTCAAGGCCTCGAGTGGATGGGTAATATCTACCCCGGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAATAGAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCACTAGGTGGACTACCGGCACAGGCGCCTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCT CGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA BAP049-Clone-B LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 54 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 97 DNA VL GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGCGGGCTACACTGAGCTGTAAATCTAGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTACTGGGCCTCTACTAGAGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACTATCTCTAGCCTGCAGCCCGAGGATATCGCTACCTACTACTGTCAGAACGACTATAGCTACCCCTACACCTTCGGTCAAGGCACTAAGGTCGAGATTAAG SEQ ID NO: 56 LC EIVLTQSPATLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGKAPKLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYTKHKNSFACE SEQ ID NO: 98 DNA LC GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGCGGGCTACACTGAGCTGTAAATCTAGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTACTGGGCCTCTACTAGAGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACTATCTCTAGCCTGCAGCCCGAGGATATCGCTACCTACTACTGTCAGAACGACTATAGCTACCCCTACACCTTCGGTCAAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC BAP049-Clone-C HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 90 DNA VH GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGAGTCCCTGCGGATCTCCTGCAAGGGCTCTGGCTACACCTTCACCACCTACTGGATGCACTGGGTGCGACAGGCTACCGGCCAGGGCCTGGAATGGATGGGCAACATCTATCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAACAGAGTGACCATCACCGCCGACAAGTCCACCTCCACCGCCTACATGGAACTGTCCTCCCTGAGATCCGAGGACACCGCCGTGTACTACTGCACCCGGTGGACAACCGGCACAGGCGCTTATTGGGGCCAGGGCACCACAGTGACCGTGTCCTCT SEQ ID NO: 91 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 92 HC DNA GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGAGTCCCTGCGGATCTCCTGCAAGGGCTCTGGCTACACCTTCACCACCTACTGGATGCACTGGGTGCGACAGGCTACCGGCCAGGGCCTGGAATGGATGGGCAACATCTATCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAACAGAGTGACCATCACCGCCGACAAGTCCACCTCCACCGCCTACATGGAACTGTCCTCCCTGAGATCCGAGGACACCGCCGTGTACTACTGCACCCGGTGGACAACCGGCACAGGCGCTTATTGGGGCCAGGGCACCACAGTGACCGTGTCCTCTGCTTCTACCAAGGGGCCCAGCGTGTTCCCCCTGGCCCCCTGCTCCAGAAGCACCAGCGAGAGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCAAGACCTACACCTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGAGCAAGTACGGCCCACCCTGCCCCCCCTGCCCAGCCCCCGAGTTCCTGGGCGGACCCAGCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGACGTGTCCCAGGAGGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTTTAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGTAAGGTCTCCAACAAGGGCCTGCCAAGCAGCATCGAAAAGACCATCA GCAAGGCCAAGGGCCAGCCTAGAGAGCCCCAGGTCTACACCCTGCCACCCAGCCAAGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCAAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAGGCTGACCGTGGACAAGTCCAGATGGCAGGAGGGCAACGTCTTTAGCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCTGGGC BAP049-Clone-C LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 66 VL EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 99 DNA VL GAGATCGTGCTGACCCAGTCCCCCGACTTCCAGTCCGTGACCCCCAAAGAAAAAGTGACCATCACATGCAAGTCCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCTACTGGGCCTCCACCCGGGAATCTGGCGTGCCCTCTAGATTCTCCGGCTCCGGCTCTGGCACCGACTTTACCTTCACCATCTCCAGCCTGGAAGCCGAGGACGCCGCCACCTACTACTGCCAGAACGACTACTCCTACCCCTACACCTTCGGCCAGGGCACCAAGGTGGAAATCAAG SEQ ID NO: 68 LC EIVLTQSPDFQSVTPKEKVTITCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQECKDSTYSLSSTLTLSKADYTKHKNSFACE SEQ ID NO: 100 DNA LC GAGATCGTGCTGACCCAGTCCCCCGACTTCCAGTCCGTGACCCCCAAAGAAAAAGTGACCATCACATGCAAGTCCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCTACTGGGCCTCCACCCGGGAATCTGGCGTGCCCTCTAGATTCTCCGGCTCCGGCTCTGGCACCGACTTTACCTTCACCATCTCCAGCCTGGAAGCCGAGGACGCCGCCACCTACTACTGCCAGAACGACTACTCCTACCCCTACACCTTCGGCCAGGGCACCAAGGTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCAAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCAGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC BAP049-Clone-D HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 50 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 101 DNA VH GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGAGTCCCTGCGGATCTCCTGCAAGGGCTCTGGCTACACCTTCACCACCTACTGGATGCACTGGATCCGGCAGTCCCCCTCTAGGGGCCTGGAATGGCTGGGCAACATCTACCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAACAGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTACCAGATGGACCACCGGAACCGGCGCCTATTGGGGCCAGGGCACAACAGTGACCGTGTCCTCC SEQ ID NO: 102 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWIRQSPSRGLEWLGNIYPGTGGSNFDEKFKNRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 103 HC DNA GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGAGTCCCTGCGGATCTCCTGCAAGGGCTCTGGCTACACCTTCACCACCTACTGGATGCACTGGATCCGGCAGTCCCCCTCTAGGGGCCTGGAATGGCTGGGCAACATCTACCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAACAGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTACCAGATGGACCACCGGAACCGGCGCCTATTGGGGCCAGGGCACAACAGTGACCGTGTCCTCCGCTTCTACCAAGGGGCCCAGCGTGTTCCCCCTGGCCCCCTGCTCCAGAAGCACCAGCGAGAGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCAAGACCTACACCTGTAACGTGGACCACAAGCCCAGCAACACCAAGGTGGACAAGAGGGTGGAGAGCAAGTACGGCCCACCCTGCCCCCCCTGCCCAGCCCCCGAGTTCCTGGGCGGACCCAGCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGACGTGTCCCAGGAGGACCCCGAGGTCCAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTTTAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGTAAGGTCTCCAACAAGGGCCTGCCAAGCAGCATCGAAAAGACCATCA GCAAGGCCAAGGGCCAGCCTAGAGAGCCCCAGGTCTACACCCTGCCACCCAGCCAAGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCAAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAGGCTGACCGTGGACAAGTCCAGATGGCAGGAGGGCAACGTCTTTAGCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCTGGGC BAP049-Clone-D LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 70 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 104 DNA VL GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCACTGTCTCCAGGCGAGAGAGCTACCCTGTCCTGCAAGTCCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCTACTGGGCCTCCACCCGGGAATCTGGCGTGCCCTCTAGATTCTCCGGCTCCGGCTCTGGCACCGACTTTACCTTCACCATCTCCAGCCTGGAAGCCGAGGACGCCGCCACCTACTACTGCCAGAACGACTACTCCTACCCCTACACCTTCGGCCAGGGCACCAAGGTGGAAATCAAG SEQ ID NO: 72 LC EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKTHNKSSVY SEQ ID NO: 105 DNA LC GAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCACTGTCTCCAGGCGAGAGAGCTACCCTGTCCTGCAAGTCCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCTACTGGGCCTCCACCCGGGAATCTGGCGTGCCCTCTAGATTCTCCGGCTCCGGCTCTGGCACCGACTTTACCTTCACCATCTCCAGCCTGGAAGCCGAGGACGCCGCCACCTACTACTGCCAGAACGACTACTCCTACCCCTACACCTTCGGCCAGGGCACCAAGGTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCAAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCAGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC BAP049-Clone-E HC SEQ ID NO: 1 (Kebat) HCDR1 TYWMH SEQ ID NO: 2 (Kebat) HCDR2 NIYPGTGGSNFDEKFKN SEQ ID NO: 3 (Kebat) HDDR3 WTTGTGAY SEQ ID NO: 4 (Chothia) HCDR1 GYTFTTY SEQ ID NO: 5 (Chothia) HCDR2 YPGTGG SEQ ID NO: 3 (Chothia) HDDR3 WTTGTGAY SEQ ID NO: 38 vh EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSS SEQ ID NO: 95 DNA VH GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCAGGCTACACCTTCACTACCTACTGGATGCACTGGGTCCGCCAGGCTACCGGTCAAGGCCTCGAGTGGATGGGTAATATCTACCCCGGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAATAGAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCACTAGGTGGACTACCGGCACAGGCGCCTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: 91 HC EVQLVQSGAEVKKPGESLRISCKGSGYTFTTYWMHWVRQATGQGLEWMGNIYPGTGGSNFDEKFKNRVTITADKSTSTAYMELSSLRSEDTAVYYCTRWTTGTGAYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 96 HC DNA GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCAGGCTACACCTTCACTACCTACTGGATGCACTGGGTCCGCCAGGCTACCGGTCAAGGCCTCGAGTGGATGGGTAATATCTACCCCGGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAATAGAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCACTAGGTGGACTACCGGCACAGGCGCCTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCT CGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA BAP049-Clone-E LC SEQ ID NO: 10 (Kebat) LCDR1 KSSQSLLDSGNQKNFLT SEQ ID NO: 11 (Kebat) LCDR2 WASTRES SEQ ID NO: 32 (Kebat) LCDR3 QNDYSYPYT SEQ ID NO: 13 (Chothia) LCDR1 SQSLLDSGNQKNF SEQ ID NO: 14 (Chothia) LCDR2 WAS SEQ ID NO: 33 (Chothia) LCDR3 DYSYPY SEQ ID NO: 70 VL EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIK SEQ ID NO: 106 DNA VL GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGCGGGCTACACTGAGCTGTAAATCTAGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGATCTACTGGGCCTCTACTAGAGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACTATCTCTAGCCTGGAAGCCGAGGACGCCGCTACCTACTACTGTCAGAACGACTATAGCTACCCCTACACCTTCGGTCAAGGCACTAAGGTCGAGATTAAG SEQ ID NO: 72 LC EIVLTQSPATLSLSPGERATLSCKSSQSLLDSGNQKNFLTWYQQKPGQAPRLLIYWASTRESGVPSRFSGSGSGTDFTFTISSLEAEDAATYYCQNDYSYPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTKTLSKADYEKTHNKSSVY SEQ ID NO: 107 DNA LC GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGCGGGCTACACTGAGCTGTAAATCTAGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACCTGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGATCTACTGGGCCTCTACTAGAGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACTATCTCTAGCCTGGAAGCCGAGGACGCCGCTACCTACTACTGTCAGAACGACTATAGCTACCCCTACACCTTCGGTCAAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC BAP049HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 115 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTGCACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 118 (Chothia) LCDR3 GATTATAGTTATCCGTGGC BAP049-chi HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-chi LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 115 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTGCACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 118 (Chothia) LCDR3 GATTATAGTTATCCGTGGC BAP049-chi Y HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-chi Y LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum01HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-HUM01LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum02HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum02LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum03HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum03LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum04HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum04LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum05HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum05LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum06HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum06LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum07HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum07LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum08HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum08LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum09HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum09LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum10HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum10LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum11HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum11LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum12HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum12LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum13HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum13LC SEQ ID NO: 121 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTAACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum14HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 223 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAC SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 223 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAC BAP049-hum14LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum15HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 223 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAC SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 223 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAC BAP049-hum15LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-hum16HC SEQ ID NO: 108 (Kebat) HCDR1 ACTTACTGGATGAC SEQ ID NO: 109 (Kebat) HCDR2 AATATTTATCCTGGTACTGGTGGTTTCTAACTTCGATGAGAAGTTCAAGAAC SEQ ID NO: 110 (Kebat) HDDR3 TGGACTACTGGGACGGGAGCTTAT SEQ ID NO: 111 (Chothia) HCDR1 GGCTACACATTCACCACTTAC SEQ ID NO: 112 (Chothia) HCDR2 TATCCTGGTACTGGTGGT SEQ ID NO: 110 (Chothia) HDDR3 TGGACTACTGGGACGGGAGCTTAT BAP049-hum16LC SEQ ID NO: 113 (Kebat) LCDR1 AAGTCCAGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTCTTGACC SEQ ID NO: 114 (Kebat) LCDR2 TGGGCATCCACTAGGGAATCT SEQ ID NO: 119 (Kebat) LCDR3 CAGAATGATTATAGTTATCCGTACACG SEQ ID NO: 116 (Chothia) LCDR1 AGTCAGAGTCTGTTAGACAGTGGAAATCAAAAGAACTTC SEQ ID NO: 117 (Chothia) LCDR2 TGGGCATCC SEQ ID NO: 120 (Chothia) LCDR3 GATTATAGTTATCCGTAC BAP049-Clone-A HC SEQ ID NO: 122 (Kebat) HCDR1 ACCTACTGGATGAC SEQ ID NO: 123 (Kebat) HCDR2 AACATCTATCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAAC SEQ ID NO: 124 (Kebat) HDDR3 TGGACAACCGGCACAGGCGCTTAT SEQ ID NO: 125 (Chothia) HCDR1 GGCTACACCTTCACCACCTAC SEQ ID NO: 126 (Chothia) HCDR2 TATCCTGGCACCGGCGGC SEQ ID NO: 124 (Chothia) HDDR3 TGGACAACCGGCACAGGCGCTTAT BAP049-Clone-A LC SEQ ID NO: 127 (Kebat) LCDR1 AAGTCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACC SEQ ID NO: 128 (Kebat) LCDR2 TGGGCCTCCACCCGGGAATCT SEQ ID NO: 129 (Kebat) LCDR3 CAGAACGACTACTCCTACCCCTACACC SEQ ID NO: 130 (Chothia) LCDR1 TCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTC SEQ ID NO: 131 (Chothia) LCDR2 TGGGCCTCC SEQ ID NO: 132 (Chothia) LCDR3 GACTACTCCTACCCCTAC BAP049-Clone-B HC SEQ ID NO: 133 (Kebat) HCDR1 ACCTACTGGATGAC SEQ ID NO: 134 (Kebat) HCDR2 AATATCTACCCCGGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAAT SEQ ID NO: 135 (Kebat) HDDR3 TGGACTACCGGCACAGGCGCCTAC SEQ ID NO: 136 (Chothia) HCDR1 GGCTACACCTTCACTACCTAC SEQ ID NO: 137 (Chothia) HCDR2 TACCCCGGCACCGGCGGC SEQ ID NO: 135 (Chothia) HDDR3 TGGACTACCGGCACAGGCGCCTAC BAP049-Clone-B LC SEQ ID NO: 138 (Kebat) LCDR1 AAATCTAGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACC SEQ ID NO: 139 (Kebat) LCDR2 TGGGCCTCTACTAGAGAATCA SEQ ID NO: 140 (Kebat) LCDR3 CAGAACGACTATAGCTACCCCTACACC SEQ ID NO: 141 (Chothia) LCDR1 AGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTC SEQ ID NO: 142 (Chothia) LCDR2 TGGGCCTCT SEQ ID NO: 143 (Chothia) LCDR3 GACTATAGCTACCCCTAC BAP049-Clone-C HC SEQ ID NO: 122 (Kebat) HCDR1 ACCTACTGGATGAC SEQ ID NO: 123 (Kebat) HCDR2 AACATCTATCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAAC SEQ ID NO: 124 (Kebat) HDDR3 TGGACAACCGGCACAGGCGCTTAT SEQ ID NO: 125 (Chothia) HCDR1 GGCTACACCTTCACCACCTAC SEQ ID NO: 126 (Chothia) HCDR2 TATCCTGGCACCGGCGGC SEQ ID NO: 124 (Chothia) HDDR3 TGGACAACCGGCACAGGCGCTTAT BAP049-Clone-C LC SEQ ID NO: 127 (Kebat) LCDR1 AAGTCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACC SEQ ID NO: 128 (Kebat) LCDR2 TGGGCCTCCACCCGGGAATCT SEQ ID NO: 129 (Kebat) LCDR3 CAGAACGACTACTCCTACCCCTACACC SEQ ID NO: 130 (Chothia) LCDR1 TCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTC SEQ ID NO: 131 (Chothia) LCDR2 TGGGCCTCC SEQ ID NO: 132 (Chothia) LCDR3 GACTACTCCTACCCCTAC BAP049-Clone-D HC SEQ ID NO: 122 (Kebat) HCDR1 ACCTACTGGATGAC SEQ ID NO: 144 (Kebat) HCDR2 AACATCTACCCTGGCACCGGCGGCTCCAACTTCGACGAGAAGTTCAAGAAC SEQ ID NO: 145 (Kebat) HDDR3 TGGACCACCGGAACCGGCGCCTAT SEQ ID NO: 125 (Chothia) HCDR1 GGCTACACCTTCACCACCTAC SEQ ID NO: 146 (Chothia) HCDR2 TACCCTGGCACCGGCGGC SEQ ID NO: 145 (Chothia) HDDR3 TGGACCACCGGAACCGGCGCCTAT BAP049-Clone-D LC SEQ ID NO: 127 (Kebat) LCDR1 AAGTCTCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTCCTGACC SEQ ID NO: 128 (Kebat) LCDR2 TGGGCCTCCACCCGGGAATCT SEQ ID NO: 129 (Kebat) LCDR3 CAGAACGACTACTCCTACCCCTACACC SEQ ID NO: 130 (Chothia) LCDR1 TCCCAGTCCCTGCTGGACTCCGGCAACCAGAAGAACTTC SEQ ID NO: 131 (Chothia) LCDR2 TGGGCCTCC SEQ ID NO: 132 (Chothia) LCDR3 GACTACTCCTACCCCTAC BAP049-Clone-E HC SEQ ID NO: 133 (Kebat) HCDR1 ACCTACTGGATGAC SEQ ID NO: 134 (Kebat) HCDR2 AATATCTACCCCGGCACCGGCGGCTCTAACTTCGACGAGAAGTTTAAGAAT SEQ ID NO: 135 (Kebat) HDDR3 TGGACTACCGGCACAGGCGCCTAC SEQ ID NO: 136 (Chothia) HCDR1 GGCTACACCTTCACTACCTAC SEQ ID NO: 137 (Chothia) HCDR2 TACCCCGGCACCGGCGGC SEQ ID NO: 135 (Chothia) HDDR3 TGGACTACCGGCACAGGCGCCTAC BAP049-Clone-E LC SEQ ID NO: 138 (Kebat) LCDR1 AAATCTAGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTCCTGACC SEQ ID NO: 139 (Kebat) LCDR2 TGGGCCTCTACTAGAGAATCA SEQ ID NO: 140 (Kebat) LCDR3 CAGAACGACTATAGCTACCCCTACACC SEQ ID NO: 141 (Chothia) LCDR1 AGTCAGTCACTGCTGGATAGCGGTAATCAGAAGAACTTC SEQ ID NO: 142 (Chothia) LCDR2 TGGGCCTCT SEQ ID NO: 143 (Chothia) LCDR3 GACTATAGCTACCCCTAC

Table 2 . Amino acid and nucleotide sequences of heavy and light chain framework regions of humanized mAbs BAP049-hum01 - BAP049-hum16 and BAP049-Clone-A - BAP049-Clone-E

Amino acid sequence Nucleotide sequence VHFW1
(type a) EVQLVQSGAEVKKPGESLRISCKGS (SEQ ID NO: 147) GAAGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAAAAGCCCGGGGAGTCTCTGAGGATCTCCTGTAAGGGTTCT (SEQ ID NO: 148)
GAAGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAGCCTGGCGAGTCCCTGCGGATCTCCTGCAAGGGCTCT (SEQ ID NO: 149)
GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCACTGAGAATTAGCTGTAAAGGTTCA (SEQ ID NO: 150) VHFW1
(type b) QVQLVQSGAEVKKPGASVKVSCKAS (SEQ ID NO: 151) CAGGTTCAGCTGGTGCAGTCTGGAGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTCTCCTGCAAGGCTTCT (SEQ ID NO: 152) VHFW2
(type a) WVRQATGQGLEWMG
(SEQ ID NO: 153) TGGGTGCGACAGGCCACTGGACAAGGGCTTGAGTGGATGGGT (SEQ ID NO: 154)
TGGGTGCGACAGGCTACCGGCCAGGGCCTGGAATGGATGGGC (SEQ ID NO: 155)
TGGGTCCGCCAGGCTACCGGTCAAGGCCTCGAGTGGATGGGT (SEQ ID NO: 156) VHFW2
(type b) WIRQSPSRGLEWLG
(SEQ ID NO: 157) TGGATCAGGCAGTCCCCATCGAGAGGCCTTGAGTGGCTGGGT (SEQ ID NO: 158)
TGGATCCGGCAGTCCCCCTCTAGGGGCCTGGAATGGCTGGGC (SEQ ID NO: 159) VHFW2
(type c) WVRQAPGQGLEWMG
(SEQ ID NO: 160) TGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGT (SEQ ID NO: 161) VHFW3
(type a) RVTITADKSTSTAYMELSSLRSEDTAVYYCTR (SEQ ID NO: 162) AGAGTCACGATTACCGCGGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTACTGTACAAGA (SEQ ID NO: 163)
AGAGTGACCATCACCGCCGACAAGTCCACCTCCACCGCCTACATGGAACTGTCCTCCCTGAGATCCGAGGACACCGCCGTGTACTACTGCACCCGG (SEQ ID NO: 164)
AGAGTGACTATCACCGCCGATAAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCACTAGG (SEQ ID NO: 165) VHFW3
(type b) RFTISRDNSKNTLYLQMNSLRAEDTAVYYCTR (SEQ ID NO: 166) AGATTCACCATCTCCAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTACAAGA (SEQ ID NO: 167)
AGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTACCAGA (SEQ ID NO: 168) VHFW4 WGQGTTVTVSS
(SEQ ID NO: 169) TGGGGCCAGGGCACCACCGTGACCGTGTCCTCC (SEQ ID NO: 170)
TGGGGCCAGGGCACCAAGTGACCGTGTCCTCT (SEQ ID NO: 171)
TGGGGTCAAGGCACTACCGTGACCGTGTCTAGC (SEQ ID NO: 172)
TGGGGCCAGGGCACAACAGTGACCGTGTCCTCC (SEQ ID NO: 173) VLFW1
(type a) EIVLTQSPDFQSVTPKEKVTITC
(SEQ ID NO: 174) GAAATTGTGCTGACTCAGTCTCAGACTTTCAGTCTGTGACTCCAAAGGAGAAAGTCACCATCACCTGC (SEQ ID NO: 175)
GAGATCGTGCTGACCAGTCCCCCGACTTCCAGTCCGTGACCCCCAAAGAAAAAGTGACCATCACATGC (SEQ ID NO: 176) VLFW1
(type b) EIVLTQSPATLSLSPGERATLSC
(SEQ ID NO: 177) GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGC (SEQ ID NO: 178)
GAGATCGTGCTGACCAGTCCCCTGCCACCCTGTCACTGTCTCCAGGCGAGAGAGCTACCCTGTCCTGC (SEQ ID NO: 179)
GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGCGGGCTACACTGAGCTGT (SEQ ID NO: 180) VLFW1
(type c) DIVMTQTPLSLPVTPGEPASISC
(SEQ ID NO: 181) GATATTGTGATGACCCAGACTCCACTCTCCCTGCCCGTCACCCCTGGAGAGCCGGCCTCCATCTCCTGC (SEQ ID NO: 182) VLFW1
(type d) DVVMTQSPLSLPVTLGQPASISC
(SEQ ID NO: 183) GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCCATCTCCTGC (SEQ ID NO: 184) VLFW1
(type e) DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO: 185) GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGC (SEQ ID NO: 186) VLFW2
(type a) WYQQKPGQAPRLLIY
(SEQ ID NO: 187) TGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTAT (SEQ ID NO: 188)
TGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCTAC (SEQ ID NO: 189)
TGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGATCTAC (SEQ ID NO: 190) VLFW2
(type b) WYQQKPGKAPKLLIY
(SEQ ID NO: 191) TGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTAT (SEQ ID NO: 192)
TGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTAC (SEQ ID NO: 193) VLFW2
(type c) WYLQKPGQSPQLLIY
(SEQ ID NO: 194) TGGTACCTGCAGAAGCCAGGGCAGTCTCCACAGCTCCTGATCTAT (SEQ ID NO: 195) VLFW3
(type a) GVPSRFSGSGSGTDFTFTISSLEAEDATYYC
(SEQ ID NO: 196) GGGGTCCCCTCGAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACCTTTACCATCAGTAGCCTGGAAGCTGAAGATGCTGCAACATATTACTGT (SEQ ID NO: 197)
GGCGTGCCCTCTAGATTCTCCGGCTCCGGCTCTGGCACCGACTTTACCTTCACCATCTCCAGCCTGGAAGCCGAGGACGCCGCCACCTACTACTGC (SEQ ID NO: 198)
GGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACTATCTCTAGCCTGGAAGCCGAGGACGCCGCTACCTACTACTGT (SEQ ID NO: 199) VLFW3
(type b) GIPPRFSGSGYGTDFTLTINNIESEDAAYYFC
(SEQ ID NO: 200) GGGATCCCACCTCGATTCAGTGGCAGCGGGTATGGAACAGATTTTACCCTCACAATTAATAACATAGAATCTGAGGATGCTGCATATTACTTCTGT (SEQ ID NO: 201) VLFW3
(type c) GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC
(SEQ ID NO: 202) GGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGT (SEQ ID NO: 203)
GGCGTGCCCTCTAGATTCTCCGGCTCCGGCTCTGGCACCGAGTTTACCCTGACCATCTCCAGCCTGCAGCCCGACGACTTCGCCACCTACTACTGC (SEQ ID NO: 204) VLFW3
(type d) GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC
(SEQ ID NO: 205) GGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGATTTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTGT (SEQ ID NO: 206)
GGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCTTCACTATCTCTAGCCTGCAGCCCGAGGATATCGCTACCTACTACTACTGT (SEQ ID NO: 207) VLFW4 FGQGTKVEIK
(SEQ ID NO: 208) TTCGGCCAAGGGACCAAGGTGGAAATCAAA (SEQ ID NO: 209)
TTCGGCCAGGGCACCAAGGTGGAAATCAAG (SEQ ID NO: 210)
TTCGGTCAAGGCACTAAGGTCGAGATTAAG (SEQ ID NO: 211)

Table 3 . Amino acid sequences of the constant region of human IgG heavy chains and human kappa light chain

HC Amino acid sequence of IgG4 (S228P) mutant constant region (EU numbering)
ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTKT YTCNVDHKPS NTKVDKRVES KYGPPCPPCP APEFLGGPSV FLFPPKPKDT LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTY RVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEG NVFSCSVMHE ALHNHYTQKS LSLSLGK (SEQ ID NO: 212) LC Amino acid sequence of the human kappa constant region
RTVAAPSVFI FPPSDEQLKS GTASVVCLLN NFYPREAKVQ WKVDNALQSG NSQESVTEQD SKDSTYSLSS TLTLSKADYE KHKVYACEVT HQGLSSPVTK
SFNRGEC (SEQ ID NO: 213) HC Amino acid sequence of IgG4 (S228P) mutant constant region without C-terminal lysine (K) (EU numbering)
ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTKT YTCNVDHKPS NTKVDKRVES KYGPPCPPCP APEFLGGPSV FLFPPKPKDT LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTY RVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTK NQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEG NVFSCSVMHE ALHNHYTQKS LSLSLG (SEQ ID NO: 214) HC IgG1 wild type
ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGG PSVFLFPPKP KDTLMISRTP EVTCVVVDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYN STYRVVSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 215) HC Amino acid sequence of IgG1 (N297A) mutant constant region (EU numbering)
ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGG PSVFLFPPKP KDTLMISRTP EVTCVVVDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYA STYRVVSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW
QQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 216) HC Amino acid sequence of IgG1 (D265A, P329A) mutant constant region (EU numbering)
ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGG PSVFLFPPKP KDTLMISRTP EVTCVVVAVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYN STYRVVSVLT VLHQDWLNGK EYKCKVSNKA LAAPIEKTIS KAKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 217) HC Amino acid sequence of IgG1 (L234A, L235A) mutant constant region (EU numbering)
ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPEAAGG PSVFLFPPKP KDTLMISRTP EVTCVVVDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYN STYRVVSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREE MTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRW QQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 218)

Table 4 . Heavy and light chain leader amino acid sequences for humanized mAbs BAP049-Clone-A - BAP049-Clone-E

BAP049-Clone-A HC MEWSWVFLFFLSVTTGVHS (SEQ ID NO: 219) LC MSVPTQVLGLLLLWLTDARC (SEQ ID NO: 220) BAP049-Clone-B HC MAWVWTLPFLMAAAQSVQA (SEQ ID NO: 221) LC MSVLTQVLALLLLWLTGTRC (SEQ ID NO: 222) BAP049-Clone-C HC MEWSWVFLFFLSVTTGVHS (SEQ ID NO: 219) LC MSVPTQVLGLLLLWLTDARC (SEQ ID NO: 220) BAP049-Clone-D HC MEWSWVFLFFLSVTTGVHS (SEQ ID NO: 219) LC MSVPTQVLGLLLLWLTDARC (SEQ ID NO: 220) BAP049-Clone-E HC MAWVWTLPFLMAAAQSVQA (SEQ ID NO: 221) LC MSVLTQVLALLLLWLTGTRC (SEQ ID NO: 222)

Table 7 . Selected therapeutic agents that can be administered in combination with anti-PD-1 antibody molecules, for example, as a single agent or in combination with other immunomodulators described in this application. Each publication listed in this Table is incorporated herein by reference in its entirety, including all structural formulas in said publication.

EXAMPLES

The examples below are provided to assist in understanding the invention, but are not intended, and should not be construed, as limiting its scope in any way.

Example 1 Analysis of the pharmacokinetics of fixed dose regimens

Based on pharmacokinetic (PK) modeling, constant dose administration is expected to result in patient exposure at appropriate Cmin concentrations. More than 99.5% of patients will have concentrations above the EC50, and more than 93% of patients will have concentrations above the EC90. The calculated mean steady state Cmin for an exemplary anti-PD-1 antibody molecule using a dose of 300 mg once every three weeks (Q3W) or 400 mg once every four weeks (Q4W) is expected to average over 20 µg/ ml (with a maximum weight of 150 kg).

Table 5 . Approximate PK parameters based on constant dose schedules

Number of patients in the PK dataset 46 CL (ml/h) 10.9 [8.9, 13.2]; IV: 62% Weight exponent on CL 0.54 [0.021, 1.06] Volume of distribution in MS (l) 7.2 [6.5, 7.9]; IV: 22% Half-life (days) 20 [17, 23]; IV: 64% Estimated Cmin (µg/ml) for a patient weighing 80 kg 31 [22, 42] (400 mg q4w)
35 [26, 47] (300 mg q3w)

Expected mean steady state Cmin concentrations for an exemplary anti-PD-1 antibody molecule observed with any dose/regimen (300mg q3w or with 400mg q4w) will be at least 77 times the EC50 (0.42µg/mL) and approximately 8.6 times the EC90. The ex vivo activity is based on the change in IL-2 in the ex vivo SEB assay.

Less than 10% of patients are expected to achieve Cmin concentrations below 3.6 µg/ml with a dose of 300 mg Q3W or 400 mg Q4W. Less than 0.5% of patients are expected to achieve Cmin concentrations below 0.4 µg/ml with a dose of 300 mg Q3W or 400 mg Q4W.

Calculated Ctrough concentrations (Cmin) for patients of various weights receiving the same dose of an exemplary anti-PD-1 antibody molecule are shown in Figure 12 . Weight-based dosing is compared to a fixed dose (3.75 mg/kg Q3W versus 300 mg Q3W and 5 mg/kg Q4W versus 400 mg Q4W). Figure 12 confirms the efficacy of administering a constant dose of an exemplary anti-PD-1 antibody molecule.

The PK model was further validated. As shown in Figure 13 , the concentrations observed and predicted by the model lie on the same line. Figures 14A-14B show that the model captures accumulation, temporal dynamics, and intra-subject variability.

Example 2: Pharmacology in vivo for combination of anti-PD-1 antibody and Compound A21

Compound A21 was combined in vivo with an exemplary anti-PD-1 antibody molecule in the MC38 syngeneic tumor model in mice. C57Bl/6 mice were implanted with 1×10 ⁶ MC38 cells/mouse. Vehicle and compound A21 (50 mg/kg po, twice a day, QW) were administered, four doses in total, starting on day 4 post-tumor implantation. The isotype and anti-mouse PD-1 antibody were administered at a dose of 10 mg/kg iv, QW.

As shown in Figure 15 , synergistic activity was observed with the combination of Compound A21 (100 mg/kg orally, once a week, administered separately, 50 mg/kg, twice a day) and anti-PD-1 antibody (10 mg/kg per /v, once a week), regardless of the effectiveness of a single agent LCL161. The combination cohort showed 4 complete responses (CR), while no CR was recorded with the introduction of a single agent.

Example 3: Pharmacology in vivo for combination of anti-PD-1 antibody and panobinostat (Compounds A19)

Panobinostat (Compound A19) was combined in vivo with an exemplary anti-PD-1 antibody molecule in a syngeneic MC38 mouse tumor model. Panobinostat (Compound A19) was administered every other day at a dose of 12 mg/kg, which resulted in drug exposure comparable to that observed in patients treated with the approved dose and regimen. Some decrease in body weight was observed with the introduction of one panobinostat and combination, with recovery after the completion of the introduction of panobinostat. As shown in Figures 16A-16B , significant synergistic activity was observed with the combination of panobinostat (Compound A19) and an anti-PD1 antibody molecule (10 mg/kg IV, once a week): all ten animals receiving the combination showed CR, compared with three out of ten animals treated with one PD-1 inhibitor or two out of ten treated with one panobinostat.

Example 4: Pharmacology in vivo for combination of anti-PD-L1 antibody and Compound A40

Compound A40 was combined in vivo with an exemplary anti-PD-L1 antibody molecule in an A20 lymphoma model. As shown in Figure 17 , the combination of anti-PD-L1 antibody and Compound A40, or anti-PD-L1 antibody and ibrutinib, was more effective than either agent alone. Compound A40 and ibrutinib were administered for only ten days, with a total of 5 doses of anti-PD L1 antibody. Even though Compound A40 and ibrutinib were administered for a short time, the survival effects of Compound A40 plus anti-PD-L1 antibody and ibrutinib plus anti-PD-L1 antibody continued after 60 days. As shown in Figure 18 , the combination of anti-PD-L1 antibody and Compound A40 also resulted in tumor regression in mice bearing A20 lymphoma allografts.

Example 5 Phase I/II Study of First Use in Humans of an Exemplary Anti-PD-1 Antibody Molecule in Patients with Advanced Solid Tumors

Antibody molecules

The exemplary antibody molecule (BAP049-Clone-E) tested in this study is a humanized anti-programmed death 1 (PD-1) monoclonal IgG4 antibody (mAb) that blocks the binding of programmed death ligand 1 (PD-L1) and programmed death ligand 1 (PD-L1). death 2 (PD-L2) with PD-1. It binds to PD-1 with high affinity and inhibits its biological activity. The amino acid sequence of this antibody molecule is described in Table 1 in this application (VH: SEQ ID NO: 38; VL: SEQ ID NO: 70). The results of preclinical toxicology studies have shown that it has a favorable safety profile. Its pharmacodynamic activity has also been demonstrated in vivo . This Example presents data from the first human administration study of this antibody molecule, phase I/II, in adults with advanced solid tumors.

Methods

Study scheme and treatment

Figure 19 shows the design of a Phase I/II multicenter, open-label study with dose escalation and expansion of dose groups to study the safety and efficacy of an exemplary antibody molecule administered to patients with advanced solid tumors.

Patients were treated with an antibody molecule until they experienced unacceptable toxicity, progressive disease according to the Modified Immune-Mediated Response Criteria in Solid Tumors ([irRC], defined as a 20% increase in the sum of the diameters of all target lesions measured. Progression should be confirmed at a second evaluation through ≥4 weeks, using as reference the smallest sum of diameters of all target lesions recorded at or after study entry (the sum must also show an absolute increase of ≥5 mm) and/or discontinued treatment at the discretion of the investigator or at their own discretion.

In the escalation phase of the study, the antibody molecule was administered intravenously (IV) every 2 weeks (Q2W) or every 4 weeks (Q4W). Five administration schedules of the antibody molecule were evaluated: 1 mg/kg Q2W, 3 mg/kg Q2W, 10 mg/kg Q2W, 3 mg/kg Q4W, and 5 mg/kg Q4W. Patients were to receive the antibody molecule until the maximum tolerated dose (MTD) was reached or until a lower recommended Phase II dose (RP2D) was established. A Bayesian linear dose-exposure model for the antibody molecule and an adaptive Bayesian logistic regression model after dose escalation with overdose control method were used to perform the dose escalation.

Study Goals and Endpoints

The objectives and endpoints of the study are shown in Table 9.

Table 9 . Study Goals and Endpoints

Goals Final indicators Primary RP2D and/or MTD assessment for an anti-PD-1 antibody molecule Exposure (AUC _[0-336h] ) after the first dose of treatment; percentage of patients with EBRT Secondary Analysis of the PK profile of an anti-PD-1 antibody molecule Serum PK values (eg, AUC, C _max , T _max , half-life); serum concentration-time profiles Safety and tolerability analysis of the anti-PD-1 antibody molecule Safety: frequency and severity of AE and SAE, including changes in laboratory parameters, vital signs, and ECG
Tolerability: interruptions, dose reductions, and dose intensity Evaluation of preliminary antitumor activity of an anti-PD-1 antibody molecule ORR, PFS, DOR and DCR Exploratory Evaluation of the pharmacodynamic effect of an antibody molecule Number of CD8+ lymphocytes infiltrating the tumor AE, adverse events; AUC, area under the curve; DCR, disease control score; DLT, dose-limiting toxicity; DOR, response duration; ECG, electrocardiogram; in / in, intravenous; ORR, percentage of patients with an objective response; PK, pharmacokinetics; PFS, progression-free survival; SAE, serious adverse events.

Key Inclusion Criteria

The study included patients with advanced/metastatic solid tumors, with measurable or non-measurable disease manifestations as defined by the Response Evaluation Criteria for Solid Tumors (RECIST) version 1.1, whose tumors progressed despite standard therapy, or who did not tolerate standard therapy, or for which there is no standard therapy. The assessment of the general condition according to the scale of the Eastern United Oncological Group was ≤2. The tumor could be biopsied, with the patient consenting to a tumor biopsy at baseline and during study drug therapy.

Main Exclusion Criteria

Patients with symptomatic central nervous system (CNS) metastases or central nervous system metastases requiring local CNS-targeted therapy (such as radiation therapy or surgery) or increased doses of corticosteroids within the previous 2 weeks were excluded from the study. Patients were excluded if they had received systemic anticancer therapy within 2 weeks of the first dose of study drug or had previously received PD-1 or PD-L1 targeted therapy. Patients requiring continuous systemic steroid therapy, other than steroid replacement doses in case of adrenal insufficiency, or who had a history of severe allergic reactions to other mAbs, were also excluded.

Ratings

Adverse events (AE) were classified using the US National Cancer Institute (NCI CTCAE) Common Terminology Criteria for Adverse Events version 4.03. Careful pharmacokinetic (PK) sampling at multiple time points (up to 336 hours post-dose Q2W and 672 hours post-dose Q4W) was performed in all Phase I patients. as a covariate for clearance and volume of distribution. X-ray response was assessed by computed tomography (CT) according to RECIST version 1.1. Scans were performed every 2 cycles from day 1 of cycle 3 to day 1 of cycle 11, then every 3 cycles until disease progression on irRC or patient withdrawal from the study. For biomarker studies, an archived tumor sample and a newly obtained pre-treatment tumor biopsy sample were collected at screening. An additional tumor biopsy was obtained during treatment.

results

Demographics and Patient Characteristics

As shown in Table 10 , 58 patients were enrolled in Phase I of the study. Patients had a wide range of advanced solid tumors. 4/58 (7%) patients had previously received 0 anticancer regimens, 7/58 (12%) had previously received 1 anticancer regimen, 13/58 (22%) had previously received 2 anticancer regimens, and 34/58 (59%) ) had previously received ≥3 anticancer regimens.

Table 10 . Demographics and Patient Characteristics

Characteristics All patients
Phase I
N=58 Median age, years (range) 55 (23-82) Gender, n (%) Women 26 (45) Men 32 (55) Race, n (%) Europeans 44 (76) African Americans 2(3) Asians 9 (16) unknown 12) Other 2(3) General condition according to the WHO/ECOG scale, n (%) 0 24 (41) 1 33 (57) 2
Diagnosis, n (%)
anal cancer
Mammary cancer
Cholangiocarcinoma
skin melanoma
Esophageal carcinoma
Stomach cancer
Head and neck cancer
Hepatocellular carcinoma
Liposarcoma
Metastatic RCC
NSCLC
prostate cancer
SCRL
TNBC
Other 12)
2(3)
2(3)
2(3)
12)
12)
12)
3(5)
2(3)
3(5)
6 (10)
12)
12)
2(3)
12)
30 (52) Number of previous anticancer regimens, n (%) 0 4 (7) 1 7(12) 2
≥3 13 (22)
34 (59) ECOG, Eastern Joint Cancer Group; NSCLC, non-small cell lung cancer; RCC, renal cell carcinoma; SCLC, small cell lung cancer; TNBC, triple-negative breast cancer; WHO, World Health Organization.

Patient distribution and exposure

As shown in Table 11 , 11/58 (19%) patients received study drug. Of the 58 patients, 46 had been in the study for >8 weeks at the end of data collection; 34 patients for >12 weeks; 20 patients for >20 weeks; 6 patients for >36 weeks. Table 11 also indicates that the main reason for discontinuing treatment was progressive disease in 39/58 (67%) patients. In Phase I patients, the median duration of exposure was 14 weeks (range 2-46); in Phase II patients, the mean duration of exposure was 2.86 weeks (range 0.6-9.9).

Table 11 . Patient distribution

1 mg/kg Q2W 3 mg/kg Q2W 10 mg/kg Q2W All Q2W patients 3 mg/kg Q4W 5 mg/kg Q4W All Q4W patients All Phase I patients N=16 N=15 N=11 N=42 N=6 N=10 N=16 N=58 Reason for distribution n(%) n(%) n(%) n(%) n(%) n(%) n(%) n(%) Patients treated Interrupted treatment 13 (81) 11 (73) 9 (82) 33 (79) 6 (100) 8 (80) 14 (88) 47 (81) Continuation of treatment* 3 (19) 4 (27) 2 (18) 9 (21) 0 2 (20) 2 (13) 11 (19) Main reason for ending treatment Adverse event sixteen) 0 0 12) 0 0 0 12) progressive disease 11 (69) 8 (53) 7 (64) 26 (62) 6 (100) 7 (70) 13 (81) 39 (67) Patient/observer decision 0 2 (13) nineteen) 3 (7) 0 0 0 3(5) Death sixteen) 1 (7) nineteen) 3 (7) 0 1 (10) sixteen) 4 (7) *Patients still participating at the time data collection was stopped.

Dose-limiting toxicity (DLT), clinical PK and RP2D

No dose-limiting toxic effects were noted. The model-predicted concentration-time profiles (Course 1/Course 3, if available; semi-log representation) for the antibody molecule as a function of dose and administration schedule are shown in Figure 14 . Peak serum concentrations (Cmax) were usually observed 1 hour after the end of the infusion. Approximately dose-proportional increases in exposure were observed with 1-10 mg/kg. The antibody molecule achieves an AUC _{of 0-336h of} approximately 1000 µg*day/mL in Course 3 at a dose of 3 mg/kg Q2W or 5 mg/kg Q4W.

Approximately 2.1-3.4-fold accumulation was observed with the Q2W dose and 1.6-2.2-fold with the Q4W dose. PK variability was low to moderate; intersubject variability (geometric mean CV%) ranged from 0.5 to 39.2% for Cmax and from 3.6 to 47.7% for AUC _0-336h (Course 1). The estimated half-life from a preliminary analysis of population PK in patients was 20 (95% CI: 17-23) days.

Based on population pharmacokinetic (PK) analysis, a constant dose of 400 mg Q4W is calculated to achieve mean steady state Ctrough concentrations of approximately 31 μg/mL (90% CI: 22-42), which exceeds the ex vivo EC50 for PD-1 blockade 0 .42 μg/ml ( Figure 12 ). Therefore, 400 mg of Q4W was chosen as the recommended dose of phase II (RP2D) for the antibody molecule. The alternative 300 mg Q3W regimen is expected to achieve similar exposure to the 400 mg Q4W dose and may be used in combination regimens where the Q3W schedule is more convenient.

Safety and tolerability

In Phase I patients, the most common (occurring in ≥20% of patients) of all classes of adverse events (AEs), regardless of relationship to study drug, were nausea, fatigue, anemia, diarrhea, dyspnea, vomiting, abdominal pain, decreased appetite, and constipation ( Table 12 ). The most common class 3/4 AE (occurring in ≥10% of patients), regardless of relationship to study drug, was anemia. The most common AE (occurring in ≥20% of patients) of all classes suspected of being related to study drug was fatigue. Class 3/4 AEs suspected of being related to study drug were rare, occurring in 3.4% of patients. No trends were observed in the adverse event profile across the dosing cohorts.

Table 12 . Adverse events regardless of interaction with investigational medicinal product (any class, observed in ≥20% of patients - safety assessment sample)

All classes Preferred Term n(%) Nausea 23 (40) Fatigue 21 (36) Anemia 19 (33) Diarrhea 17 (29) Dyspnea 17 (29) Vomit 14 (24) Abdominal pain 13 (22) Decreased appetite 13 (22) Constipation 12 (21) Only adverse events reported during treatment or within 90 days of the last drug administration in the study are reported.

In Phase II patients, the most common (observed in ≥10% of patients) of all AE classes, regardless of association with study drug, were abdominal pain, constipation, cough, decreased appetite, dyspnea, fatigue, and elevated gamma-glutamyltransferase. The most common class 3/4 AEs (seen in ≥5% of patients), regardless of study drug association, were abdominal pain and elevated gamma-glutamyltransferase. The most common AEs (observed in ≥5% of patients) of all classes suspected of being associated with study drug were fatigue, nausea, and pruritus. Neither recorded any AE 3/4 suspected of being related to study drug.

No infusion reactions or dose reductions were noted. One patient discontinued treatment due to an adverse event (Class 3, dyspnea) not suspected of being related to study drug.

Efficiency

The overall response rate and disease control rate for the various tumor types were 2% and 41%, respectively ( Table 13 ). Changes in tumor mass over time for each of the patients to be x-rayed can be seen in Figure 20 . Diagnosis of patients remaining in the study at the end of Phase I data collection includes: liposarcoma (two patients), testicular cancer, atypical lung carcinoid tumor, anaplastic thyroid carcinoma, Merkel cell carcinoma, clear cell renal cell carcinoma, melanoma, and triple negative breast cancer (TNBC). As shown in Figures 21A-21B , a partial response after pseudoprogression was observed in one patient with metastatic atypical pulmonary carcinoid. As shown in Figure 21C , high levels of CD8+ T lymphocytes were detected by immunohistochemical staining in a tumor sample obtained from this patient on Day 1 of Course 2.

Table 13 . Best overall response (based on investigator's assessment of disease status using RECIST v1.1 criteria)

1 mg/kg Q2W
(N=16)
n(%) 3 mg/kg Q2W
(N=15)
n(%) 10 mg/kg Q2W
(N=11)
n(%) All Q2W patients
(N=42)
n(%) 3 mg/kg Q4W
(N=6)
n(%) 5 mg/kg Q4W
(N=10)
n(%) All Q4W patients
(N=16)
n(%) All Phase I patients
(N=58)
n(%) Eligible Patients* sixteen 15 eleven 42 6 10 sixteen 58 Best Overall Answer Complete response (CR) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) Partial Response (PR) 0 (0) 1 (7) 0 (0) 12) 0 (0) 0 (0) 0 (0) 12) Stable disease (SD) 9 (56) 6 (40) 3 (27) 18 (43) 3 (50) 2 (20) 5 (31) 23 (40) Progressive Disease (PD) 5 (31) 6 (40) 4 (36) 15 (36) 3 (50) 7 (70) 10 (63) 25 (43) unknown 2 (13) 2 (13) 4 (36) 8 (19) 0 (0) 1 (10) sixteen) 9 (16) Total Patient Response Rate (CR or PR) 0 (0) 1 (7) 0 (0) 12) 0 (0) 0 (0) 0 (0) 12) 90% confidence interval* [0; 17] [0; 28] [0; 24] [0; eleven] [0; 39] [0; 26] [0; 17] [0; 8] Disease control score (CR or PR or SD) 9 (56) 7 (47) 3 (27) 19 (45) 3 (50) 2 (20) 5 (31) 24 (41) 90% confidence interval [33; 77] [24; 70] [8; 56] [32; 59.0] [15; 85] [four; 51] [13; 55] [thirty; 53] SD includes patients with best overall non-CR/non-PD response; A 90% confidence interval was calculated using the exact interval (Clopper-Pearson).
*Eligible patients are defined as patients with at least one post-treatment assessment or who discontinued treatment prior to the first assessment after enrollment in the study.

findings

The exemplary antibody molecule was well tolerated and had a controlled safety profile. Chose RP2D 400 mg Q4W. The alternative dosing schedule of 300 mg Q3W is expected to achieve similar exposure and may be a more convenient alternative for evaluating some combination therapy regimens that include an antibody molecule. Preliminary efficacy data in one patient suggest an antitumor response driven by increased CD8+ T-lymphocyte activity, as expected for a clinically active PD-1 inhibitor.

Example 6 Pharmacology in vivo for combination of anti-PD-1 antibody and Compound A15

Compound A15 was tested in combination with an exemplary anti-PD-1 antibody molecule in the MC38 syngeneic colorectal cancer model selected for the presence of TAM and anti-PD-1 antibody response. As shown in Figures 22A-22C , the combination of anti-PD-1 antibody and Compound A15 inhibited tumor growth and increased survival compared to anti-PD-1 antibody alone. In this study, weekly administration of Compound A15 (200 mg/kg) in combination with an anti-PD-1 antibody molecule was more effective (8 out of 11 complete responders) than daily administration of Compound A15 (200 mg/kg) in combination with weekly dosing anti-PD-1 antibody molecule (4 out of 11 complete responders) and one anti-PD-1 antibody molecule (5 out of 11 complete responders, weekly dosing). A moderate decrease in TAM was noted 2 days after the second dose in the Compound A15 weekly regimen. The observed TAM kinetics in the weekly combination regimen of Compound A15 and anti-PD-1 antibody was transient as TAM returned to normal values 7 days after the second dose. A decrease in tumor Tregs was observed at all time points analyzed (2 and 7 days post-dose) with Compound A15 plus an anti-PD-1 antibody molecule in a weekly dosing regimen.

Example 7 Anti-PD-L1 Antibody Activity in MC38 Colon Cancer Model in Combination with Anti-PD-1 Antibody

The combination of an exemplary anti-PD-1 antibody molecule and an exemplary anti-PD-L1 antibody molecule was tested in a mouse model of colon adenocarcinoma. A surrogate anti-murine PD-1 antibody, RMP1-14, was used. As shown in Figure 23 , co-administration of RMP1-14 and an anti-PD-L1 antibody molecule resulted in enhanced antitumor activity compared to both agents alone in this model. In particular, the anti-PD-L1 and RMP1-14 antibody molecule in combination resulted in 4/10 animals with a complete response, and also 3/10 animals showed partial responses. In addition, combination therapy resulted in a non-significant increase in median time to end point to 46 days, 21 days longer than isotype-treated animals and 20 days longer than antibody alone.

Other embodiments and examples, including figures and tables, are disclosed in International Patent Application Publication WO 2015/112900 and US Patent Application Publication 2015/0210769 entitled "Antibody Molecules to PD-1 and Uses Thereof", which are incorporated by reference in their entirety. .

INCLUSION BY REFERENCE

All publications, patents, and registration numbers referenced herein are incorporated by reference in their entirety, as if each individual publication or patent were expressly and individually identified as being incorporated by reference.

EQUIVALENTS

While certain embodiments of the present invention have been discussed, the above description is illustrative and non-limiting. Various variations of the invention will become apparent to those skilled in the art upon reading the present description and the claims below. The full scope of the invention is to be determined by reference to the claims, along with their full scope of equivalents, and the description, along with such variations.

--->

SEQUENCE LIST

<110> NOVARTIS AG

<120> PD-1 ANTIBODY MOLECULES AND THEIR APPLICATIONS

<130> C2160-7015WO

<140>

<141>

<150> 62/431.846

<151> 2016-12-09

<150> 62/414.128

<151> 2016-10-28

<150> 62/400.787

<151> 2016-09-28

<150> 62/381.384

<151> 2016-08-30

<150> 62/359.781

<151> 2016-07-08

<150> 62/347,331

<151> 2016-06-08

<150> 62/344.784

<151> 2016-06-02

<150> 62/331.371

<151> 2016-05-03

<150> 62/269.044

<151> 2015-12-17

<160> 327

<170>PatentIn version 3.5

<210> 1

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 1

Thr Tyr Trp Met His

15

<210> 2

<211> 17

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 2

Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe Lys

1 5 10 15

Asn

<210> 3

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 3

Trp Thr Thr Gly Thr Gly Ala Tyr

15

<210> 4

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 4

Gly Tyr Thr Phe Thr Thr Tyr

15

<210> 5

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 5

Tyr Pro Gly Thr Gly Gly

15

<210> 6

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 6

Gln Val Gln Leu Gln Gln Pro Gly Ser Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Thr Ser Leu Thr Val Asp Thr Ser Ser Thr Thr Ala Tyr

65 70 75 80

Met His Leu Ala Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ala

115

<210> 7

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 7

caggtccagc tgcagcaacc tgggtctgag ctggtgaggc ctggagcttc agtgaagctg 60

tcctgcaagg cgtctggcta cacattcacc acttactgga tgcactggggt gaggcagagg 120

cctggacaag gccttgagtg gattggaaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaaaaacag gacctcactg actgtagaca catcctccac cacagcctac 240

atgcacctcg ccagcctgac atctgaggac tctgcggtct attactgtac aagatggact 300

actgggacgg gagcttattg gggccaaggg actctggtca ctgtctctgc a 351

<210> 8

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 8

Gln Val Gln Leu Gln Gln Ser Gly Ser Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Thr Ser Leu Thr Val Asp Thr Ser Ser Thr Thr Ala Tyr

65 70 75 80

Met His Leu Ala Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ala

115

<210> 9

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 9

caggtccagc tgcagcagtc tgggtctgag ctggtgaggc ctggagcttc agtgaagctg 60

tcctgcaagg cgtctggcta cacattcacc acttactgga tgcactggggt gaggcagagg 120

cctggacaag gccttgagtg gattggaaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaaaaacag gacctcactg actgtagaca catcctccac cacagcctac 240

atgcacctcg ccagcctgac atctgaggac tctgcggtct attactgtac aagatggact 300

actgggacgg gagcttattg gggccaaggg actctggtca ctgtctctgc a 351

<210> 10

<211> 17

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 10

Lys Ser Ser Gln Ser Leu Leu Asp Ser Gly Asn Gln Lys Asn Phe Leu

1 5 10 15

Thr

<210> 11

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 11

Trp Ala Ser Thr Arg Glu Ser

15

<210> 12

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 12

Gln Asn Asp Tyr Ser Tyr Pro Cys Thr

15

<210> 13

<211> 13

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 13

Ser Gln Ser Leu Leu Asp Ser Gly Asn Gln Lys Asn Phe

1 5 10

<210> 14

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 14

Trp Ala Ser

1

<210> 15

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 15

Asp Tyr Ser Tyr Pro Cys

15

<210> 16

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 16

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Val Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Cys Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile

100 105 110

Lys

<210> 17

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 17

gacattgtga tgacccagtc tccatcctcc ctgactgtga cagcaggaga gaaggtcact 60

atgagctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaaccagg gcagcctcct aaactgttga tcttctgggc atccactagg 180

gaatctgggg tccctgatcg cttcacaggc agtggatctg taacagattt cactctcacc 240

atcagcagtg tgcaggctga agacctggca gtttattact gtcagaatga ttatagttat 300

ccgtgcacgt tcggaggggg gaccaagctg gaaataaaa 339

<210> 18

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 18

Gln Val Gln Leu Gln Gln Pro Gly Ser Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Thr Ser Leu Thr Val Asp Thr Ser Ser Thr Thr Ala Tyr

65 70 75 80

Met His Leu Ala Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser

115

<210> 19

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 19

caggtccagc tgcagcagcc tgggtctgag ctggtgaggc ctggagcttc agtgaagctg 60

tcctgcaagg cgtctggcta cacattcacc acttactgga tgcactggggt gaggcagagg 120

cctggacaag gccttgagtg gattggaaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaaaaacag gacctcactg actgtagaca catcctccac cacagcctac 240

atgcacctcg ccagcctgac atctgaggac tctgcggtct attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc c 351

<210> 20

<211> 444

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 20

Gln Val Gln Leu Gln Gln Pro Gly Ser Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Thr Ser Leu Thr Val Asp Thr Ser Ser Thr Thr Ala Tyr

65 70 75 80

Met His Leu Ala Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 21

<211> 1332

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 21

caggtccagc tgcagcagcc tgggtctgag ctggtgaggc ctggagcttc agtgaagctg 60

tcctgcaagg cgtctggcta cacattcacc acttactgga tgcactggggt gaggcagagg 120

cctggacaag gccttgagtg gattggaaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaaaaacag gacctcactg actgtagaca catcctccac cacagcctac 240

atgcacctcg ccagcctgac atctgaggac tctgcggtct attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc cgcttccacc 360

aagggcccat ccgtcttccc cctggcgccc tgctccagga gcacctccga gagcacagcc 420

gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacgaagac ctacacctgc 600

aacgtagatc acaagcccag caacaccaag gtggacaaga gagttgagtc caaatatggt 660

cccccatgcc caccgtgccc agcacctgag ttcctggggg gaccatcagt cttcctgttc 720

cccccaaaac ccaaggacac tctcatgatc tcccggaccc ctgaggtcac gtgcgtggtg 780

gtggacgtga gccaggaaga ccccgaggtc cagttcaact ggtacgtgga tggcgtggag 840

gtgcataatg ccaagacaaa gccgcgggag gagcagttca acagcacgta ccgtgtggtc 900

agcgtcctca ccgtcctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 960

tccaacaaag gcctcccgtc ctccatcgag aaaaccatct ccaaagccaa agggcagccc 1020

cgagagccac aggtgtacac cctgccccca tcccaggagg agatgaccaa gaaccaggtc 1080

agcctgacct gcctggtcaa aggcttctac cccagcgaca tcgccgtgga gtgggagagc 1140

aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc 1200

ttcttcctct acagcaggct aaccgtggac aagagcaggt ggcaggaggg gaatgtcttc 1260

tcatgctccg tgatgcatga ggctctgcac aaccactaca cacagaagag cctctccctg 1320

tctctgggta aa 1332

<210> 22

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 22

Gln Val Gln Leu Gln Gln Ser Gly Ser Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Thr Ser Leu Thr Val Asp Thr Ser Ser Thr Thr Ala Tyr

65 70 75 80

Met His Leu Ala Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser

115

<210> 23

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 23

caggtccagc tgcagcagtc tgggtctgag ctggtgaggc ctggagcttc agtgaagctg 60

tcctgcaagg cgtctggcta cacattcacc acttactgga tgcactggggt gaggcagagg 120

cctggacaag gccttgagtg gattggaaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaaaaacag gacctcactg actgtagaca catcctccac cacagcctac 240

atgcacctcg ccagcctgac atctgaggac tctgcggtct attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc c 351

<210> 24

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 24

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Val Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Cys Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 25

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 25

gacattgtga tgacccagtc tccatcctcc ctgactgtga cagcaggaga gaaggtcact 60

atgagctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaaccagg gcagcctcct aaactgttga tcttctgggc atccactagg 180

gaatctgggg tccctgatcg cttcacaggc agtggatctg taacagattt cactctcacc 240

atcagcagtg tgcaggctga agacctggca gtttattact gtcagaatga ttatagttat 300

ccgtgcacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 26

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 26

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Val Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Cys Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 27

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 27

gacattgtga tgacccagtc tccatcctcc ctgactgtga cagcaggaga gaaggtcact 60

atgagctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaaccagg gcagcctcct aaactgttga tcttctgggc atccactagg 180

gaatctgggg tccctgatcg cttcacaggc agtggatctg taacagattt cactctcacc 240

atcagcagtg tgcaggctga agacctggca gtttattact gtcagaatga ttatagttat 300

ccgtgcacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 28

<400> 28

000

<210> 29

<400> 29

000

<210> 30

<211> 444

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 30

Gln Val Gln Leu Gln Gln Ser Gly Ser Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Thr Ser Leu Thr Val Asp Thr Ser Ser Thr Thr Ala Tyr

65 70 75 80

Met His Leu Ala Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 31

<211> 1332

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 31

caggtccagc tgcagcagtc tgggtctgag ctggtgaggc ctggagcttc agtgaagctg 60

tcctgcaagg cgtctggcta cacattcacc acttactgga tgcactggggt gaggcagagg 120

cctggacaag gccttgagtg gattggaaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaaaaacag gacctcactg actgtagaca catcctccac cacagcctac 240

atgcacctcg ccagcctgac atctgaggac tctgcggtct attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc cgcttccacc 360

aagggcccat ccgtcttccc cctggcgccc tgctccagga gcacctccga gagcacagcc 420

gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacgaagac ctacacctgc 600

aacgtagatc acaagcccag caacaccaag gtggacaaga gagttgagtc caaatatggt 660

cccccatgcc caccgtgccc agcacctgag ttcctggggg gaccatcagt cttcctgttc 720

cccccaaaac ccaaggacac tctcatgatc tcccggaccc ctgaggtcac gtgcgtggtg 780

gtggacgtga gccaggaaga ccccgaggtc cagttcaact ggtacgtgga tggcgtggag 840

gtgcataatg ccaagacaaa gccgcgggag gagcagttca acagcacgta ccgtgtggtc 900

agcgtcctca ccgtcctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 960

tccaacaaag gcctcccgtc ctccatcgag aaaaccatct ccaaagccaa agggcagccc 1020

cgagagccac aggtgtacac cctgccccca tcccaggagg agatgaccaa gaaccaggtc 1080

agcctgacct gcctggtcaa aggcttctac cccagcgaca tcgccgtgga gtgggagagc 1140

aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc 1200

ttcttcctct acagcaggct aaccgtggac aagagcaggt ggcaggaggg gaatgtcttc 1260

tcatgctccg tgatgcatga ggctctgcac aaccactaca cacagaagag cctctccctg 1320

tctctgggta aa 1332

<210> 32

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 32

Gln Asn Asp Tyr Ser Tyr Pro Tyr Thr

15

<210> 33

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 33

Asp Tyr Ser Tyr Pro Tyr

15

<210> 34

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 34

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Val Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 35

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 35

gacattgtga tgacccagtc tccatcctcc ctgactgtga cagcaggaga gaaggtcact 60

atgagctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaaccagg gcagcctcct aaactgttga tcttctgggc atccactagg 180

gaatctgggg tccctgatcg cttcacaggc agtggatctg taacagattt cactctcacc 240

atcagcagtg tgcaggctga agacctggca gtttattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 36

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 36

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Val Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 37

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 37

gacattgtga tgacccagtc tccatcctcc ctgactgtga cagcaggaga gaaggtcact 60

atgagctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaaccagg gcagcctcct aaactgttga tcttctgggc atccactagg 180

gaatctgggg tccctgatcg cttcacaggc agtggatctg taacagattt cactctcacc 240

atcagcagtg tgcaggctga agacctggca gtttattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 38

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 38

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser

115

<210> 39

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 39

gaagtgcagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60

tcctgtaagg gttctggcta cacattcacc acttactgga tgcactgggt gcgacaggcc 120

actggacaag ggcttgagtg gatgggtaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaagaacag agtcacgatt accgcggaca aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc c 351

<210> 40

<211> 444

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 40

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 41

<211> 1332

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 41

gaagtgcagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60

tcctgtaagg gttctggcta cacattcacc acttactgga tgcactgggt gcgacaggcc 120

actggacaag ggcttgagtg gatgggtaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaagaacag agtcacgatt accgcggaca aatccacgag cacagcctac 240

atggagctga gcagcctgag atctgaggac acggccgtgt attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc cgcttccacc 360

aagggcccat ccgtcttccc cctggcgccc tgctccagga gcacctccga gagcacagcc 420

gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacgaagac ctacacctgc 600

aacgtagatc acaagcccag caacaccaag gtggacaaga gagttgagtc caaatatggt 660

cccccatgcc caccgtgccc agcacctgag ttcctggggg gaccatcagt cttcctgttc 720

cccccaaaac ccaaggacac tctcatgatc tcccggaccc ctgaggtcac gtgcgtggtg 780

gtggacgtga gccaggaaga ccccgaggtc cagttcaact ggtacgtgga tggcgtggag 840

gtgcataatg ccaagacaaa gccgcgggag gagcagttca acagcacgta ccgtgtggtc 900

agcgtcctca ccgtcctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 960

tccaacaaag gcctcccgtc ctccatcgag aaaaccatct ccaaagccaa agggcagccc 1020

cgagagccac aggtgtacac cctgccccca tcccaggagg agatgaccaa gaaccaggtc 1080

agcctgacct gcctggtcaa aggcttctac cccagcgaca tcgccgtgga gtgggagagc 1140

aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc 1200

ttcttcctct acagcaggct aaccgtggac aagagcaggt ggcaggaggg gaatgtcttc 1260

tcatgctccg tgatgcatga ggctctgcac aaccactaca cacagaagag cctctccctg 1320

tctctgggta aa 1332

<210> 42

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 42

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 43

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 43

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctgggg tcccatcaag gttcagcggc agtggatctg ggacagaatt cactctcacc 240

atcagcagcc tgcagcctga tgattttgca acttattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 44

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 44

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 45

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 45

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctgggg tcccatcaag gttcagcggc agtggatctg ggacagaatt cactctcacc 240

atcagcagcc tgcagcctga tgattttgca acttattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 46

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 46

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Ile

50 55 60

Pro Pro Arg Phe Ser Gly Ser Gly Tyr Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Asn Asn Ile Glu Ser Glu Asp Ala Ala Tyr Tyr Phe Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 47

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 47

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctggga tcccacctcg attcagtggc agcgggtatg gaacagattt taccctcaca 240

attaataaca tagaatctga ggatgctgca tattacttct gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 48

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 48

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Ile

50 55 60

Pro Pro Arg Phe Ser Gly Ser Gly Tyr Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Asn Asn Ile Glu Ser Glu Asp Ala Ala Tyr Tyr Phe Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 49

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 49

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctggga tcccacctcg attcagtggc agcgggtatg gaacagattt taccctcaca 240

attaataaca tagaatctga ggatgctgca tattacttct gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 50

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 50

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser

115

<210> 51

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 51

gaagtgcagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60

tcctgtaagg gttctggcta cacattcacc acttactgga tgcactggat caggcagtcc 120

ccatcgagag gccttgagtg gctgggtaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaagaacag attcaccatc tccagagaca attccaagaa cacgctgtat 240

cttcaaatga acagcctgag agccgaggac acggccgtgt attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc c 351

<210> 52

<211> 444

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 52

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 53

<211> 1332

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 53

gaagtgcagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60

tcctgtaagg gttctggcta cacattcacc acttactgga tgcactggat caggcagtcc 120

ccatcgagag gccttgagtg gctgggtaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaagaacag attcaccatc tccagagaca attccaagaa cacgctgtat 240

cttcaaatga acagcctgag agccgaggac acggccgtgt attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc cgcttccacc 360

aagggcccat ccgtcttccc cctggcgccc tgctccagga gcacctccga gagcacagcc 420

gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacgaagac ctacacctgc 600

aacgtagatc acaagcccag caacaccaag gtggacaaga gagttgagtc caaatatggt 660

cccccatgcc caccgtgccc agcacctgag ttcctggggg gaccatcagt cttcctgttc 720

cccccaaaac ccaaggacac tctcatgatc tcccggaccc ctgaggtcac gtgcgtggtg 780

gtggacgtga gccaggaaga ccccgaggtc cagttcaact ggtacgtgga tggcgtggag 840

gtgcataatg ccaagacaaa gccgcgggag gagcagttca acagcacgta ccgtgtggtc 900

agcgtcctca ccgtcctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 960

tccaacaaag gcctcccgtc ctccatcgag aaaaccatct ccaaagccaa agggcagccc 1020

cgagagccac aggtgtacac cctgccccca tcccaggagg agatgaccaa gaaccaggtc 1080

agcctgacct gcctggtcaa aggcttctac cccagcgaca tcgccgtgga gtgggagagc 1140

aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc 1200

ttcttcctct acagcaggct aaccgtggac aagagcaggt ggcaggaggg gaatgtcttc 1260

tcatgctccg tgatgcatga ggctctgcac aaccactaca cacagaagag cctctccctg 1320

tctctgggta aa 1332

<210> 54

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 54

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 55

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 55

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtatcagc agaaaccagg gaaagctcct aagctcctga tctattgggc atccactagg 180

gaatctgggg tcccatcaag gttcagtgga agtggatctg ggacagattt tactttcacc 240

atcagcagcc tgcagcctga agatattgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 56

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 56

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 57

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 57

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtatcagc agaaaccagg gaaagctcct aagctcctga tctattgggc atccactagg 180

gaatctgggg tcccatcaag gttcagtgga agtggatctg ggacagattt tactttcacc 240

atcagcagcc tgcagcctga agatattgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 58

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 58

Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 59

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 59

gatattgtga tgacccagac tccactctcc ctgcccgtca cccctggaga gccggcctcc 60

atctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 60

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 60

Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 61

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 61

gatattgtga tgacccagac tccactctcc ctgcccgtca cccctggaga gccggcctcc 60

atctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 62

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 62

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 63

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 63

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtatcagc agaaaccagg gaaagctcct aagctcctga tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 64

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 64

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 65

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 65

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtatcagc agaaaccagg gaaagctcct aagctcctga tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 66

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 66

Glu Ile Val Leu Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys

1 5 10 15

Glu Lys Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 67

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 67

gaaattgtgc tgactcagtc tccagacttt cagtctgtga ctccaaagga gaaagtcacc 60

atcacctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 68

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 68

Glu Ile Val Leu Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys

1 5 10 15

Glu Lys Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 69

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 69

gaaattgtgc tgactcagtc tccagacttt cagtctgtga ctccaaagga gaaagtcacc 60

atcacctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 70

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 70

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 71

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 71

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 72

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 72

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ala Pro Arg Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 73

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 73

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 74

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 74

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Leu Gln Lys Pro Gly Gln

35 40 45

Ser Pro Gln Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 75

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 75

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtacctgc agaagccagg gcagtctcca cagctcctga tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 76

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 76

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Leu Gln Lys Pro Gly Gln

35 40 45

Ser Pro Gln Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 77

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 77

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttgacc 120

tggtacctgc agaagccagg gcagtctcca cagctcctga tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 78

<211> 113

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 78

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys

<210> 79

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 79

gatgttgtga tgactcagtc tccactctcc ctgcccgtca cccttggaca gccggcctcc 60

atctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttaacc 120

tggtatcagc agaaaccagg gaaagctcct aagctcctga tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaa 339

<210> 80

<211> 220

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 80

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Lys

35 40 45

Ala Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr

65 70 75 80

Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 81

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 81

gatgttgtga tgactcagtc tccactctcc ctgcccgtca cccttggaca gccggcctcc 60

atctcctgca agtccagtca gagtctgtta gacagtggaa atcaaaagaa cttcttaacc 120

tggtatcagc agaaaccagg gaaagctcct aagctcctga tctattgggc atccactagg 180

gaatctgggg tcccctcgag gttcagtggc agtggatctg ggacagattt cacctttacc 240

atcagtagcc tggaagctga agatgctgca acatattact gtcagaatga ttatagttat 300

ccgtacacgt tcggccaagg gaccaaggtg gaaatcaaac gtacggtggc tgcaccatct 360

gtcttcatct tcccgccatc tgatgagcag ttgaaatctg gaactgcctc tgttgtgtgc 420

ctgctgaata acttctatcc cagaggcc aaagtacagt ggaaggtgga taacgccctc 480

caatcgggta actccgga gagtgtcaca gagcaggaca gcaaggacag cacctacagc 540

ctcagcagca ccctgacgct gagcaaagca gactacgaga aacacaaagt ctacgcctgc 600

gaagtcaccc atcagggcct gagctcgccc gtcacaaaga gcttcaacag gggagagtgt 660

<210> 82

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 82

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser

115

<210> 83

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 83

caggttcagc tggtgcagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggcta cacattcacc acttactgga tgcactggat caggcagtcc 120

ccatcgagag gccttgagtg gctgggtaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaagaacag attcaccatc tccagagaca attccaagaa cacgctgtat 240

cttcaaatga acagcctgag agccgaggac acggccgtgt attactgtac aagatggact 300

actgggacgg gagcttactg gggccagggc accaccgtga ccgtgtcctc c 351

<210> 84

<211> 444

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 84

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 85

<211> 1332

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 85

caggttcagc tggtgcagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttctggcta cacattcacc acttactgga tgcactggat caggcagtcc 120

ccatcgagag gccttgagtg gctgggtaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaagaacag attcaccatc tccagagaca attccaagaa cacgctgtat 240

cttcaaatga acagcctgag agccgaggac acggccgtgt attactgtac aagatggact 300

actgggacgg gagcttactg gggccagggc accaccgtga ccgtgtcctc cgcttccacc 360

aagggcccat ccgtcttccc cctggcgccc tgctccagga gcacctccga gagcacagcc 420

gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacgaagac ctacacctgc 600

aacgtagatc acaagcccag caacaccaag gtggacaaga gagttgagtc caaatatggt 660

cccccatgcc caccgtgccc agcacctgag ttcctggggg gaccatcagt cttcctgttc 720

cccccaaaac ccaaggacac tctcatgatc tcccggaccc ctgaggtcac gtgcgtggtg 780

gtggacgtga gccaggaaga ccccgaggtc cagttcaact ggtacgtgga tggcgtggag 840

gtgcataatg ccaagacaaa gccgcgggag gagcagttca acagcacgta ccgtgtggtc 900

agcgtcctca ccgtcctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 960

tccaacaaag gcctcccgtc ctccatcgag aaaaccatct ccaaagccaa agggcagccc 1020

cgagagccac aggtgtacac cctgccccca tcccaggagg agatgaccaa gaaccaggtc 1080

agcctgacct gcctggtcaa aggcttctac cccagcgaca tcgccgtgga gtgggagagc 1140

aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc 1200

ttcttcctct acagcaggct aaccgtggac aagagcaggt ggcaggaggg gaatgtcttc 1260

tcatgctccg tgatgcatga ggctctgcac aaccactaca cacagaagag cctctccctg 1320

tctctgggta aa 1332

<210> 86

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 86

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser

115

<210> 87

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 87

gaagtgcagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60

tcctgtaagg gttctggcta cacattcacc acttactgga tgcactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggtaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaagaacag attcaccatc tccagagaca attccaagaa cacgctgtat 240

cttcaaatga acagcctgag agccgaggac acggccgtgt attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc c 351

<210> 88

<211> 444

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 88

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 89

<211> 1332

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 89

gaagtgcagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60

tcctgtaagg gttctggcta cacattcacc acttactgga tgcactgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggtaat atttatcctg gtactggtgg ttctaacttc 180

gatgagaagt tcaagaacag attcaccatc tccagagaca attccaagaa cacgctgtat 240

cttcaaatga acagcctgag agccgaggac acggccgtgt attactgtac aagatggact 300

actgggacgg gagcttattg gggccagggc accaccgtga ccgtgtcctc cgcttccacc 360

aagggcccat ccgtcttccc cctggcgccc tgctccagga gcacctccga gagcacagcc 420

gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 480

ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac 540

tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacgaagac ctacacctgc 600

aacgtagatc acaagcccag caacaccaag gtggacaaga gagttgagtc caaatatggt 660

cccccatgcc caccgtgccc agcacctgag ttcctggggg gaccatcagt cttcctgttc 720

cccccaaaac ccaaggacac tctcatgatc tcccggaccc ctgaggtcac gtgcgtggtg 780

gtggacgtga gccaggaaga ccccgaggtc cagttcaact ggtacgtgga tggcgtggag 840

gtgcataatg ccaagacaaa gccgcgggag gagcagttca acagcacgta ccgtgtggtc 900

agcgtcctca ccgtcctgca ccaggactgg ctgaacggca aggagtacaa gtgcaaggtg 960

tccaacaaag gcctcccgtc ctccatcgag aaaaccatct ccaaagccaa agggcagccc 1020

cgagagccac aggtgtacac cctgccccca tcccaggagg agatgaccaa gaaccaggtc 1080

agcctgacct gcctggtcaa aggcttctac cccagcgaca tcgccgtgga gtgggagagc 1140

aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc cgacggctcc 1200

ttcttcctct acagcaggct aaccgtggac aagagcaggt ggcaggaggg gaatgtcttc 1260

tcatgctccg tgatgcatga ggctctgcac aaccactaca cacagaagag cctctccctg 1320

tctctgggta aa 1332

<210> 90

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 90

gaagtgcagc tggtgcagtc tggcgccgaa gtgaagaagc ctggcgagtc cctgcggatc 60

tcctgcaagg gctctggcta caccttcacc acctactgga tgcactgggt gcgacaggct 120

accggccagg gcctggaatg gatgggcaac atctatcctg gcaccggcgg ctccaacttc 180

gacgagaagt tcaagaacag agtgaccatc accgccgaca agtccacctc caccgcctac 240

atggaactgt ccctccctgag atccgaggac accgccgtgt actactgcac ccggtggaca 300

accggcacag gcgcttattg gggccagggc accacagtga ccgtgtcctc t 351

<210> 91

<211> 443

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 91

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440

<210> 92

<211> 1329

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 92

gaagtgcagc tggtgcagtc tggcgccgaa gtgaagaagc ctggcgagtc cctgcggatc 60

tcctgcaagg gctctggcta caccttcacc acctactgga tgcactggggt gcgacaggct 120

accggccagg gcctggaatg gatgggcaac atctatcctg gcaccggcgg ctccaacttc 180

gacgagaagt tcaagaacag agtgaccatc accgccgaca agtccacctc caccgcctac 240

atggaactgt ccctccctgag atccgaggac accgccgtgt actactgcac ccggtggaca 300

accggcacag gcgcttattg gggccagggc accacagtga ccgtgtcctc tgcttctacc 360

aaggggccca gcgtgttccc cctggccccc tgctccagaa gcaccagcga gagcacagcc 420

gccctgggct gcctggtgaa ggactacttc cccgagcccg tgaccgtgtc ctggaacagc 480

ggagccctga ccagcggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac cgtgcccagc agcagcctgg gcaccaagac ctacacctgt 600

aacgtggacc acaagcccag caacaccaag gtggacaaga gggtggagag caagtacggc 660

ccaccctgcc ccccctgccc agcccccgag ttcctgggcg gacccagcgt gttcctgttc 720

ccccccaagc ccaaggacac cctgatgatc agcagaaccc ccgaggtgac ctgtgtggtg 780

gtggacgtgt cccaggagga ccccgaggtc cagttcaact ggtacgtgga cggcgtggag 840

gtgcacaacg ccaagaccaa gcccagagag gagcagttta acagcaccta cggggtggtg 900

tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aagagtacaa gtgtaaggtc 960

tccaacaagg gcctgccaag cagcatcgaa aagaccatca gcaaggccaa gggccagcct 1020

agagagcccc aggtctacac cctgccaccc agccaagagg agatgaccaa gaaccaggtg 1080

tccctgacct gtctggtgaa gggcttctac ccaagcgaca tcgccgtgga gtgggagagc 1140

aacggccagc ccgagaacaa ctacaagacc acccccccag tgctggacag cgacggcagc 1200

ttcttcctgt acagcaggct gaccgtggac aagtccagat ggcaggaggg caacgtcttt 1260

agctgctccg tgatgcacga ggccctgcac aaccactaca cccagaagag cctgagcctg 1320

tccctgggc 1329

<210> 93

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 93

gagatcgtgc tgacccagtc ccctgccacc ctgtcactgt ctccaggcga gagagctacc 60

ctgtcctgca agtcctccca gtccctgctg gactccggca accagaagaa cttcctgacc 120

tggtatcagc agaagcccgg ccaggccccc agactgctga tctactgggc ctccacccgg 180

gaatctggcg tgccctctag attctccggc tccggctctg gcaccgagtt taccctgacc 240

atctccagcc tgcagcccga cgacttcgcc acctactact gccagaacga ctactcctac 300

ccctacacct tcggccaggg caccaaggtg gaaatcaag 339

<210> 94

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 94

gagatcgtgc tgacccagtc ccctgccacc ctgtcactgt ctccaggcga gagagctacc 60

ctgtcctgca agtcctccca gtccctgctg gactccggca accagaagaa cttcctgacc 120

tggtatcagc agaagcccgg ccaggccccc agactgctga tctactgggc ctccacccgg 180

gaatctggcg tgccctctag attctccggc tccggctctg gcaccgagtt taccctgacc 240

atctccagcc tgcagcccga cgacttcgcc acctactact gccagaacga ctactcctac 300

ccctacacct tcggccaggg caccaaggtg gaaatcaagc gtacggtggc cgctcccagc 360

gtgttcatct tccccccaag cgacgagcag ctgaagagcg gcaccgccag cgtggtgtgt 420

ctgctgaaca acttctaccc cagggaggcc aaggtgcagt ggaaggtgga caacgccctg 480

cagagcggca acagccagga gagcgtcacc gagcaggaca gcaaggactc cacctacagc 540

ctgagcagca ccctgaccct gagcaaggcc gactacgaga agcacaaggt gtacgcctgt 600

gaggtgaccc accagggcct gtccagcccc gtgaccaaga gcttcaacag gggcgagtgc 660

<210> 95

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 95

gaggtgcagc tggtgcagtc aggcgccgaa gtgaagaagc ccggcgagtc actgagaatt 60

agctgtaaag gttcaggcta caccttcact acctactgga tgcactgggt ccgccaggct 120

accggtcaag gcctcgagtg gatgggtaat atctaccccg gcaccggcgg ctctaacttc 180

240 gacgagaagt ttaagaatag agtgactatc accgccgata

atggaactgt ctagcctgag atcagaggac accgccgtct actactgcac taggtggact 300

accggcacag gcgcctactg gggtcaaggc actaccgtga ccgtgtctag c 351

<210> 96

<211> 1329

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 96

gaggtgcagc tggtgcagtc aggcgccgaa gtgaagaagc ccggcgagtc actgagaatt 60

agctgtaaag gttcaggcta caccttcact acctactgga tgcactgggt ccgccaggct 120

accggtcaag gcctcgagtg gatgggtaat atctaccccg gcaccggcgg ctctaacttc 180

240 gacgagaagt ttaagaatag agtgactatc accgccgata

atggaactgt ctagcctgag atcagaggac accgccgtct actactgcac taggtggact 300

accggcacag gcgcctactg gggtcaaggc actaccgtga ccgtgtctag cgctagcact 360

aagggcccgt ccgtgttccc cctggcacct tgtagccgga gcactagcga atccaccgct 420

gccctcggct gcctggtcaa ggattacttc ccggagcccg tgaccgtgtc ctggaacagc 480

ggagccctga cctccggagt gcacaccttc cccgctgtgc tgcagagctc cgggctgtac 540

tcgctgtcgt cggtggtcac ggtgccttca tctagcctgg gtaccaagac ctacacttgc 600

aacgtggacc acaagccttc caacactaag gtggacaagc gcgtcgaatc gaagtacggc 660

ccaccgtgcc cgccttgtcc cgcgccggag ttcctcggcg gtccctcggt ctttctgttc 720

ccaccgaagc ccaaggacac tttgatgatt tcccgcaccc ctgaagtgac atgcgtggtc 780

840

gtgcacaacg ccaaaaccaa gccgagggag gagcagttca actccactta ccgcgtcgtg 900

tccgtgctga cggtgctgca tcaggactgg ctgaacggga aggagtacaa gtgcaaagtg 960

tccaacaagg gacttcctag ctcaatcgaa aagaccatct cgaaagccaa gggacagccc 1020

cgggaacccc aagtgtatac cctgccaccg agcggaag aaatgactaa gaaccaagtc 1080

tcattgactt gccttgtgaa gggcttctac ccatcggata tcgccgtgga atgggagtcc 1140

aacggccagc cggaaaacaa ctacaagacc acccctccgg tgctggactc agacggatcc 1200

ttcttcctct actcgcggct gaccgtggat aagagcagat ggcaggaggg aaatgtgttc 1260

agctgttctg tgatgcatga agccctgcac aaccactaca ctcagaagtc cctgtccctc 1320

tccctggga 1329

<210> 97

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 97

gagatcgtcc tgactcagtc acccgctacc ctgagcctga gccctggcga gcgggctaca 60

ctgagctgta aatctagtca gtcactgctg gatagcggta atcagaagaa cttcctgacc 120

tggtatcagc agaagcccgg taaagcccct aagctgctga tctactgggc ctctactaga 180

gaatcaggcg tgccctctag gtttagcggt agcggtagtg gcaccgactt caccttcact 240

atctctagcc tgcagcccga ggatatcgct acctactact gtcagaacga ctatagctac 300

ccctacacct tcggtcaagg cactaaggtc gagattaag 339

<210> 98

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 98

gagatcgtcc tgactcagtc acccgctacc ctgagcctga gccctggcga gcgggctaca 60

ctgagctgta aatctagtca gtcactgctg gatagcggta atcagaagaa cttcctgacc 120

tggtatcagc agaagcccgg taaagcccct aagctgctga tctactgggc ctctactaga 180

gaatcaggcg tgccctctag gtttagcggt agcggtagtg gcaccgactt caccttcact 240

atctctagcc tgcagcccga ggatatcgct acctactact gtcagaacga ctatagctac 300

ccctacacct tcggtcaagg cactaaggtc gagattaagc gtacggtggc cgctcccagc 360

gtgttcatct tcccccccag cgacgagcag ctgaagagcg gcaccgccag cgtggtgtgc 420

ctgctgaaca acttctaccc ccgggaggcc aaggtgcagt ggaaggtgga caacgccctg 480

cagagcggca acagccagga gagcgtcacc gagcaggaca gcaaggactc cacctacagc 540

ctgagcagca ccctgaccct gagcaaggcc gactacgaga agcataaggt gtacgcctgc 600

gaggtgaccc accagggcct gtccagcccc gtgaccaaga gcttcaacag gggcgagtgc 660

<210> 99

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 99

gagatcgtgc tgacccagtc ccccgacttc cagtccgtga cccccaaaga aaaagtgacc 60

atcacatgca agtcctccca gtccctgctg gactccggca accagaagaa cttcctgacc 120

tggtatcagc agaagcccgg ccaggccccc agactgctga tctactgggc ctccacccgg 180

gaatctggcg tgccctctag attctccggc tccggctctg gcaccgactt taccttcacc 240

atctccagcc tggaagccga ggacgccgcc acctactact gccagaacga ctactcctac 300

ccctacacct tcggccaggg caccaaggtg gaaatcaag 339

<210> 100

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 100

gagatcgtgc tgacccagtc ccccgacttc cagtccgtga cccccaaaga aaaagtgacc 60

atcacatgca agtcctccca gtccctgctg gactccggca accagaagaa cttcctgacc 120

tggtatcagc agaagcccgg ccaggccccc agactgctga tctactgggc ctccacccgg 180

gaatctggcg tgccctctag attctccggc tccggctctg gcaccgactt taccttcacc 240

atctccagcc tggaagccga ggacgccgcc acctactact gccagaacga ctactcctac 300

ccctacacct tcggccaggg caccaaggtg gaaatcaagc gtacggtggc cgctcccagc 360

gtgttcatct tccccccaag cgacgagcag ctgaagagcg gcaccgccag cgtggtgtgt 420

ctgctgaaca acttctaccc cagggaggcc aaggtgcagt ggaaggtgga caacgccctg 480

cagagcggca acagccagga gagcgtcacc gagcaggaca gcaaggactc cacctacagc 540

ctgagcagca ccctgaccct gagcaaggcc gactacgaga agcacaaggt gtacgcctgt 600

gaggtgaccc accagggcct gtccagcccc gtgaccaaga gcttcaacag gggcgagtgc 660

<210> 101

<211> 351

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 101

gaagtgcagc tggtgcagtc tggcgccgaa gtgaagaagc ctggcgagtc cctgcggatc 60

tcctgcaagg gctctggcta caccttcacc acctactgga tgcactggat ccggcagtcc 120

ccctctaggg gcctggaatg gctgggcaac atctaccctg gcaccggcgg ctccaacttc 180

gacgagaagt tcaagaacag gttcaccatc tcccgggaca actccaagaa caccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtac cagatggacc 300

accggaaccg gcgcctattg gggccagggc acaacagtga ccgtgtcctc c 351

<210> 102

<211> 443

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 102

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Thr

100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu

115 120 125

Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn

195 200 205

Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro

210 215 220

Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

290 295 300

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440

<210> 103

<211> 1329

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 103

gaagtgcagc tggtgcagtc tggcgccgaa gtgaagaagc ctggcgagtc cctgcggatc 60

tcctgcaagg gctctggcta caccttcacc acctactgga tgcactggat ccggcagtcc 120

ccctctaggg gcctggaatg gctgggcaac atctaccctg gcaccggcgg ctccaacttc 180

gacgagaagt tcaagaacag gttcaccatc tcccgggaca actccaagaa caccctgtac 240

ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgtac cagatggacc 300

accggaaccg gcgcctattg gggccagggc acaacagtga ccgtgtcctc cgcttctacc 360

aaggggccca gcgtgttccc cctggccccc tgctccagaa gcaccagcga gagcacagcc 420

gccctgggct gcctggtgaa ggactacttc cccgagcccg tgaccgtgtc ctggaacagc 480

ggagccctga ccagcggcgt gcacaccttc cccgccgtgc tgcagagcag cggcctgtac 540

agcctgagca gcgtggtgac cgtgcccagc agcagcctgg gcaccaagac ctacacctgt 600

aacgtggacc acaagcccag caacaccaag gtggacaaga gggtggagag caagtacggc 660

ccaccctgcc ccccctgccc agcccccgag ttcctgggcg gacccagcgt gttcctgttc 720

ccccccaagc ccaaggacac cctgatgatc agcagaaccc ccgaggtgac ctgtgtggtg 780

gtggacgtgt cccaggagga ccccgaggtc cagttcaact ggtacgtgga cggcgtggag 840

gtgcacaacg ccaagaccaa gcccagagag gagcagttta acagcaccta cggggtggtg 900

tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aagagtacaa gtgtaaggtc 960

tccaacaagg gcctgccaag cagcatcgaa aagaccatca gcaaggccaa gggccagcct 1020

agagagcccc aggtctacac cctgccaccc agccaagagg agatgaccaa gaaccaggtg 1080

tccctgacct gtctggtgaa gggcttctac ccaagcgaca tcgccgtgga gtgggagagc 1140

aacggccagc ccgagaacaa ctacaagacc acccccccag tgctggacag cgacggcagc 1200

ttcttcctgt acagcaggct gaccgtggac aagtccagat ggcaggaggg caacgtcttt 1260

agctgctccg tgatgcacga ggccctgcac aaccactaca cccagaagag cctgagcctg 1320

tccctgggc 1329

<210> 104

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 104

gagatcgtgc tgacccagtc ccctgccacc ctgtcactgt ctccaggcga gagagctacc 60

ctgtcctgca agtcctccca gtccctgctg gactccggca accagaagaa cttcctgacc 120

tggtatcagc agaagcccgg ccaggccccc agactgctga tctactgggc ctccacccgg 180

gaatctggcg tgccctctag attctccggc tccggctctg gcaccgactt taccttcacc 240

atctccagcc tggaagccga ggacgccgcc acctactact gccagaacga ctactcctac 300

ccctacacct tcggccaggg caccaaggtg gaaatcaag 339

<210> 105

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 105

gagatcgtgc tgacccagtc ccctgccacc ctgtcactgt ctccaggcga gagagctacc 60

ctgtcctgca agtcctccca gtccctgctg gactccggca accagaagaa cttcctgacc 120

tggtatcagc agaagcccgg ccaggccccc agactgctga tctactgggc ctccacccgg 180

gaatctggcg tgccctctag attctccggc tccggctctg gcaccgactt taccttcacc 240

atctccagcc tggaagccga ggacgccgcc acctactact gccagaacga ctactcctac 300

ccctacacct tcggccaggg caccaaggtg gaaatcaagc gtacggtggc cgctcccagc 360

gtgttcatct tccccccaag cgacgagcag ctgaagagcg gcaccgccag cgtggtgtgt 420

ctgctgaaca acttctaccc cagggaggcc aaggtgcagt ggaaggtgga caacgccctg 480

cagagcggca acagccagga gagcgtcacc gagcaggaca gcaaggactc cacctacagc 540

ctgagcagca ccctgaccct gagcaaggcc gactacgaga agcacaaggt gtacgcctgt 600

gaggtgaccc accagggcct gtccagcccc gtgaccaaga gcttcaacag gggcgagtgc 660

<210> 106

<211> 339

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 106

gagatcgtcc tgactcagtc acccgctacc ctgagcctga gccctggcga gcgggctaca 60

ctgagctgta aatctagtca gtcactgctg gatagcggta atcagaagaa cttcctgacc 120

tggtatcagc agaagcccgg tcaagcccct agactgctga tctactgggc ctctactaga 180

gaatcaggcg tgccctctag gtttagcggt agcggtagtg gcaccgactt caccttcact 240

atctctagcc tggaagccga ggacgccgct acctactact gtcagaacga ctatagctac 300

ccctacacct tcggtcaagg cactaaggtc gagattaag 339

<210> 107

<211> 660

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polynucleotide"

<400> 107

gagatcgtcc tgactcagtc acccgctacc ctgagcctga gccctggcga gcgggctaca 60

ctgagctgta aatctagtca gtcactgctg gatagcggta atcagaagaa cttcctgacc 120

tggtatcagc agaagcccgg tcaagcccct agactgctga tctactgggc ctctactaga 180

gaatcaggcg tgccctctag gtttagcggt agcggtagtg gcaccgactt caccttcact 240

atctctagcc tggaagccga ggacgccgct acctactact gtcagaacga ctatagctac 300

ccctacacct tcggtcaagg cactaaggtc gagattaagc gtacggtggc cgctcccagc 360

gtgttcatct tcccccccag cgacgagcag ctgaagagcg gcaccgccag cgtggtgtgc 420

ctgctgaaca acttctaccc ccgggaggcc aaggtgcagt ggaaggtgga caacgccctg 480

cagagcggca acagccagga gagcgtcacc gagcaggaca gcaaggactc cacctacagc 540

ctgagcagca ccctgaccct gagcaaggcc gactacgaga agcataaggt gtacgcctgc 600

gaggtgaccc accagggcct gtccagcccc gtgaccaaga gcttcaacag gggcgagtgc 660

<210> 108

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 108

acttactgga tgcac 15

<210> 109

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 109

aatattattc ctggtactgg tggttctaac ttcgatgaga agttcaagaa c 51

<210> 110

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 110

tggactactg ggacgggagc ttat 24

<210> 111

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 111

ggctacacat tcaccactta c 21

<210> 112

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 112

tatcctggta ctggtggt 18

<210> 113

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 113

aagtccagtc agagtctgtt agacagtgga aatcaaaaga acttcttgac c 51

<210> 114

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 114

tgggcatcca ctagggaatc t 21

<210> 115

<211> 27

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 115

cagaatgatt atagttatcc gtgcacg 27

<210> 116

<211> 39

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 116

agtcagagtc tgttagacag tggaaatcaa aagaacttc 39

<210> 117

<211> 9

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 117

tgggcatcc 9

<210> 118

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 118

gattatagtt atccgtgc 18

<210> 119

<211> 27

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 119

cagaatgatt atagttatcc gtacacg 27

<210> 120

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 120

gattatagtt atccgtac 18

<210> 121

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 121

aagtccagtc agagtctgtt agacagtgga aatcaaaaga acttcttaac c 51

<210> 122

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 122

acctactgga tgcac 15

<210> 123

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 123

aacatctatc ctggcaccgg cggctccaac ttcgacgaga agttcaagaa c 51

<210> 124

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 124

tggacaaccg gcacaggcgc ttat 24

<210> 125

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 125

ggctacacct tcaccaccta c 21

<210> 126

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 126

tatcctggca ccggcggc 18

<210> 127

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 127

aagtcctccc agtccctgct ggactccggc aaccagaaga acttcctgac c 51

<210> 128

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 128

tgggcctcca cccgggaatc t 21

<210> 129

<211> 27

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 129

cagaacgact actcctaccc ctacacc 27

<210> 130

<211> 39

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 130

tcccagtccc tgctggactc cggcaaccag aagaacttc 39

<210> 131

<211> 9

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 131

tgggcctcc 9

<210> 132

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 132

gactactcct acccctac 18

<210> 133

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 133

acctactgga tgcac 15

<210> 134

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 134

aatatctacc ccggcaccgg cggctctaac ttcgacgaga agtttaagaa t 51

<210> 135

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 135

tggactaccg gcacaggcgc ctac 24

<210> 136

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 136

ggctacacct tcactaccta c 21

<210> 137

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 137

taccccggca ccggcggc 18

<210> 138

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 138

aaatctagtc agtcactgct ggatagcggt aatcagaaga acttcctgac c 51

<210> 139

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 139

tgggcctcta ctagagaatc a 21

<210> 140

<211> 27

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 140

cagaacgact atagctaccc ctacacc 27

<210> 141

<211> 39

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 141

agtcagtcac tgctggatag cggtaatcag aagaacttc 39

<210> 142

<211> 9

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 142

tgggcctct 9

<210> 143

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 143

gactatagct acccctac 18

<210> 144

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 144

aacatctacc ctggcaccgg cggctccaac ttcgacgaga agttcaagaa c 51

<210> 145

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 145

tggaccaccg gaaccggcgc ctat 24

<210> 146

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 146

taccctggca ccggcggc 18

<210> 147

<211> 25

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 147

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu

1 5 10 15

Ser Leu Arg Ile Ser Cys Lys Gly Ser

20 25

<210> 148

<211> 75

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 148

gaagtgcagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaggatc 60

tcctgtaagg gttct 75

<210> 149

<211> 75

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 149

gaagtgcagc tggtgcagtc tggcgccgaa gtgaagaagc ctggcgagtc cctgcggatc 60

tcctgcaagg gctct 75

<210> 150

<211> 75

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 150

gaggtgcagc tggtgcagtc aggcgccgaa gtgaagaagc ccggcgagtc actgagaatt 60

agctgtaaag gttca 75

<210> 151

<211> 25

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 151

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser

20 25

<210> 152

<211> 75

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 152

caggttcagc tggtgcagtc tggagctgag gtgaagaagc ctggggcctc agtgaaggtc 60

tcctgcaagg cttct 75

<210> 153

<211> 14

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 153

Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met Gly

1 5 10

<210> 154

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 154

tgggtgcgac aggccactgg acaagggctt gagtggatgg gt 42

<210> 155

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 155

tgggtgcgac aggctaccgg ccagggcctg gaatggatgg gc 42

<210> 156

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 156

tgggtccgcc aggctaccgg tcaaggcctc gagtggatgg gt 42

<210> 157

<211> 14

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 157

Trp Ile Arg Gln Ser Pro Ser Arg Gly Leu Glu Trp Leu Gly

1 5 10

<210> 158

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 158

tggatcaggc agtccccatc gagaggcctt gagtggctgg gt 42

<210> 159

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 159

tggatccggc agtccccctc taggggcctg gaatggctgg gc 42

<210> 160

<211> 14

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 160

Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly

1 5 10

<210> 161

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 161

tgggtgcgac aggcccctgg acaagggctt gagtggatgg gt 42

<210> 162

<211> 32

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 162

Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr Met Glu

1 5 10 15

Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg

20 25 30

<210> 163

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 163

agagtcacga ttaccgcgga caaatccacg agcacagcct acatggagct gagcagcctg 60

agatctgagg acacggccgt gtattactgt acaaga 96

<210> 164

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 164

agagtgacca tcaccgccga caagtccacc tccaccgcct acatggaact gtcctccctg 60

agatccgagg acaccgccgt gtactactgc acccgg 96

<210> 165

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 165

agagtgacta tcaccgccga taagtctact agcaccgcct atatggaact gtctagcctg 60

agatcagagg acaccgccgt ctactactgc actagg 96

<210> 166

<211> 32

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 166

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

1 5 10 15

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Thr Arg

20 25 30

<210> 167

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 167

agattcacca tctccagaga caattccaag aacacgctgt atcttcaaat gaacagcctg 60

agagccgagg acacggccgt gtattactgt acaaga 96

<210> 168

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 168

aggttcacca tctcccggga caactccaag aacaccctgt acctgcagat gaactccctg 60

cgggccgagg acaccgccgt gtactactgt accaga 96

<210> 169

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 169

Trp Gly Glyn Gly Thr Thr Val Thr Val Ser Ser

1 5 10

<210> 170

<211> 33

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 170

tggggccagg gcaccaccgt gaccgtgtcc tcc 33

<210> 171

<211> 33

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 171

tggggccagg gcaccacagt gaccgtgtcc tct 33

<210> 172

<211> 33

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 172

tggggtcaag gcactaccgt gaccgtgtct agc 33

<210> 173

<211> 33

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 173

tggggccagg gcacaacagt gaccgtgtcc tcc 33

<210> 174

<211> 23

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 174

Glu Ile Val Leu Thr Gln Ser Pro Asp Phe Gln Ser Val Thr Pro Lys

1 5 10 15

Glu Lys Val Thr Ile Thr Cys

20

<210> 175

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 175

gaaattgtgc tgactcagtc tccagacttt cagtctgtga ctccaaagga gaaagtcacc 60

atcacctgc 69

<210> 176

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 176

gagatcgtgc tgacccagtc ccccgacttc cagtccgtga cccccaaaga aaaagtgacc 60

atcacatgc 69

<210> 177

<211> 23

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 177

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys

20

<210> 178

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 178

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60

ctctcctgc 69

<210> 179

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 179

gagatcgtgc tgacccagtc ccctgccacc ctgtcactgt ctccaggcga gagagctacc 60

ctgtcctgc 69

<210> 180

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 180

gagatcgtcc tgactcagtc acccgctacc ctgagcctga gccctggcga gcgggctaca 60

ctgagctgt 69

<210> 181

<211> 23

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 181

Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys

20

<210> 182

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 182

gatattgtga tgacccagac tccactctcc ctgcccgtca cccctggaga gccggcctcc 60

atctcctgc 69

<210> 183

<211> 23

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 183

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys

20

<210> 184

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 184

gatgttgtga tgactcagtc tccactctcc ctgcccgtca cccttggaca gccggcctcc 60

atctcctgc 69

<210> 185

<211> 23

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 185

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys

20

<210> 186

<211> 69

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 186

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgc 69

<210> 187

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 187

Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr

1 5 10 15

<210> 188

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 188

tggtaccagc agaaacctgg ccaggctccc aggctcctca tctat 45

<210> 189

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 189

tggtatcagc agaagcccgg ccaggccccc agactgctga tctac 45

<210> 190

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 190

tggtatcagc agaagcccgg tcaagcccct agactgctga tctac 45

<210> 191

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 191

Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 192

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 192

tggtatcagc agaaaccagg gaaagctcct aagctcctga tctat 45

<210> 193

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 193

tggtatcagc agaagcccgg taaagcccct aagctgctga tctac 45

<210> 194

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 194

Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr

1 5 10 15

<210> 195

<211> 45

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 195

tggtacctgc agaagccagg gcagtctcca cagctcctga tctat 45

<210> 196

<211> 32

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 196

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Phe Thr Ile Ser Ser Leu Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys

20 25 30

<210> 197

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 197

ggggtcccct cgaggttcag tggcagtgga tctgggacag atttcacctt taccatcagt 60

agcctggaag ctgaagatgc tgcaacatat tactgt 96

<210> 198

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 198

ggcgtgccct ctagattctc cggctccggc tctggcaccg actttacctt caccatctcc 60

agcctggaag ccgaggacgc cgccacctac tactgc 96

<210> 199

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 199

ggcgtgccct ctaggtttag cggtagcggt agtggcaccg acttcacctt cactatctct 60

agcctggaag ccgaggacgc cgctacctac tactgt 96

<210> 200

<211> 32

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 200

Gly Ile Pro Pro Arg Phe Ser Gly Ser Gly Tyr Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Asn Asn Ile Glu Ser Glu Asp Ala Ala Tyr Tyr Phe Cys

20 25 30

<210> 201

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 201

gggatcccac ctcgattcag tggcagcggg tatggaacag attttaccct cacaattaat 60

aacatagaat ctgaggatgc tgcatattac ttctgt 96

<210> 202

<211> 32

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 202

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Pro Asp Asp Phe Ala Thr Tyr Tyr Cys

20 25 30

<210> 203

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 203

ggggtcccat caaggttcag cggcagtgga tctgggacag aattcactct caccatcagc 60

agcctgcagc ctgatgattt tgcaacttat tactgt 96

<210> 204

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 204

ggcgtgccct ctagattctc cggctccggc tctggcaccg agtttaccct gaccatctcc 60

agcctgcagc ccgacgactt cgccacctac tactgc 96

<210> 205

<211> 32

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 205

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys

20 25 30

<210> 206

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 206

ggggtcccat caaggttcag tggaagtgga tctgggacag attttacttt caccatcagc 60

agcctgcagc ctgaagatat tgcaacatat tactgt 96

<210> 207

<211> 96

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 207

ggcgtgccct ctaggtttag cggtagcggt agtggcaccg acttcacctt cactatctct 60

agcctgcagc ccgaggatat cgctacctac tactgt 96

<210> 208

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 208

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

1 5 10

<210> 209

<211> 30

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 209

ttcggccaag ggaccaaggt ggaaatcaaa 30

<210> 210

<211> 30

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 210

ttcggccagg gcaccaaggt ggaaatcaag 30

<210> 211

<211> 30

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 211

ttcggtcaag gcactaaggt cgagattaag 30

<210> 212

<211> 327

<212> Protein

<213> Homo sapiens

<400> 212

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro

100 105 110

Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

115 120 125

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

130 135 140

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155 160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200 205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr Lys

225 230 235 240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250 255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265 270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280 285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295 300

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

305 310 315 320

Leu Ser Leu Ser Leu Gly Lys

325

<210> 213

<211> 107

<212> Protein

<213> Homo sapiens

<400> 213

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 214

<211> 326

<212> Protein

<213> Homo sapiens

<400> 214

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro

100 105 110

Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

115 120 125

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

130 135 140

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155 160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200 205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr Lys

225 230 235 240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250 255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265 270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280 285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295 300

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

305 310 315 320

Leu Ser Leu Ser Leu Gly

325

<210> 215

<211> 330

<212> Protein

<213> Homo sapiens

<400> 215

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 216

<211> 330

<212> Protein

<213> Homo sapiens

<400> 216

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 217

<211> 330

<212> Protein

<213> Homo sapiens

<400> 217

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Ala Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Ala Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 218

<211> 330

<212> Protein

<213> Homo sapiens

<400> 218

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 219

<211> 19

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 219

Met Glu Trp Ser Trp Val Phe Leu Phe Phe Leu Ser Val Thr Thr Gly

1 5 10 15

Val His Ser

<210> 220

<211> 20

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 220

Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr

1 5 10 15

Asp Ala Arg Cys

20

<210> 221

<211> 19

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 221

Met Ala Trp Val Trp Thr Leu Pro Phe Leu Met Ala Ala Ala Gln Ser

1 5 10 15

Val Gln Ala

<210> 222

<211> 20

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 222

Met Ser Val Leu Thr Gln Val Leu Ala Leu Leu Leu Leu Trp Leu Thr

1 5 10 15

Gly Thr Arg Cys

20

<210> 223

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<400> 223

tggactactg ggacgggagc ttac 24

<210> 224

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 224

Gly Tyr Thr Phe Thr Thr Tyr Trp Met His

1 5 10

<210> 225

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 225

Cys Asn Gly Arg Cys

15

<210> 226

<400> 226

000

<210> 227

<400> 227

000

<210> 228

<211> 134

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 228

Gln Val Gln Leu Gln Gln Pro Gly Ser Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Tyr

20 25 30

Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Gly Thr Gly Gly Ser Asn Phe Asp Glu Lys Phe

50 55 60

Lys Asn Arg Thr Ser Leu Thr Val Asp Thr Ser Ser Thr Thr Ala Tyr

65 70 75 80

Met His Leu Ala Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Trp Thr Thr Gly Thr Gly Ala Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro Leu

115 120 125

Ala Pro Gly Ser Ala Ala

130

<210> 229

<211> 116

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 229

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser

20 25 30

Gly Asn Gln Lys Asn Phe Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Phe Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Val Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro Cys Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile

100 105 110

Lys Arg Ala Asp

115

<210> 230

<211> 98

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 230

Gln Val Gln Leu Gln Gln Pro Gly Ser Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met His Trp Val Lys Gln Arg His Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asn Ile Tyr Pro Gly Ser Gly Ser Thr Asn Tyr Asp Glu Lys Phe

50 55 60

Lys Ser Lys Gly Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met His Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Arg

<210> 231

<211> 101

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 231

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly

1 5 10 15

Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser

20 25 30

Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn

85 90 95

Asp Tyr Ser Tyr Pro

one hundred

<210> 232

<211> 37

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<220>

<221> CDS

<222> (2)..(37)

<400> 232

g tgc acg ttc gga ggg ggg acc aag ctg gaa ata aaa 37

Cys Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 233

<211> 12

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 233

Cys Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 234

<211> 38

<212> DNA

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

oligonucleotide"

<220>

<221> CDS

<222> (2)..(37)

<400> 234

g tac acg ttc gga ggg ggg acc aag ctg gaa ata aaa c 38

Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 235

<211> 12

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 235

Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

1 5 10

<210> 236

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 236

Met Tyr Pro Pro Tyr

15

<210> 237

<211> 4

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 237

Arg Gly Asp Ser

1

<210> 238

<211> 447

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 238

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 239

<211> 215

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 239

Glu Thr Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Leu Gly Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ala Asp Ser Pro

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 240

<211> 121

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 240

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Leu Trp Glu Val Arg Ala Leu Pro Ser Val Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 241

<211> 109

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 241

Ser Tyr Glu Leu Thr Gln Pro Ser Val Ser Val Ala Pro Gly Gln

1 5 10 15

Thr Ala Arg Ile Thr Cys Gly Ala Asn Asp Ile Gly Ser Lys Ser Val

20 25 30

His Trp Tyr Gln Gln Lys Ala Gly Gln Ala Pro Val Leu Val Val Ser

35 40 45

Glu Asp Ile Ile Arg Pro Ser Gly Ile Pro Glu Arg Ile Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Val Glu Ala Gly

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Arg Asp Ser Asp Gln

85 90 95

Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105

<210> 242

<211> 440

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 242

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr Phe Ser Asn Ser

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser

115 120 125

Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp

130 135 140

Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr

145 150 155 160

Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Ser Gly Leu Tyr

165 170 175

Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys

180 185 190

Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp

195 200 205

Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala

210 215 220

Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro

225 230 235 240

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

245 250 255

Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val

260 265 270

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

275 280 285

Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln

290 295 300

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly

305 310 315 320

Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro

325 330 335

Arg Glu Pro Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr

340 345 350

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

355 360 365

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

370 375 380

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr

385 390 395 400

Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe

405 410 415

Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys

420 425 430

Ser Leu Ser Leu Ser Leu Gly Lys

435 440

<210> 243

<211> 214

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 243

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser Asn Trp Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 244

<211> 447

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 244

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 245

<211> 218

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 245

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser

20 25 30

Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ser Arg

85 90 95

Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 246

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 246

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln

65 70 75 80

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg

85 90 95

Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 247

<211> 110

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 247

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser

85 90 95

Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 248

<211> 253

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 248

Met Ala Pro Arg Arg Ala Arg Gly Cys Arg Thr Leu Gly Leu Pro Ala

1 5 10 15

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

20 25 30

Lys Asp Asp Asp Asp Lys Ile Glu Gly Arg Ile Thr Cys Pro Pro Pro

35 40 45

Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr

50 55 60

Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly

65 70 75 80

Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala

85 90 95

His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val

100 105 110

His Gln Arg Pro Ala Pro Pro Ser Gly Gly Ser Gly Gly Gly Gly Ser

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Leu Gln Asn Trp Val Asn Val

130 135 140

Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile

145 150 155 160

Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val

165 170 175

Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu

180 185 190

Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu

195 200 205

Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys

210 215 220

Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln

225 230 235 240

Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser

245 250

<210> 249

<211> 263

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 249

Met Asp Ser Lys Gly Ser Ser Gln Lys Ala Gly Ser Arg Leu Leu Leu

1 5 10 15

Leu Leu Val Val Ser Asn Leu Leu Leu Cys Gln Gly Val Val Ser Thr

20 25 30

Thr Arg Asp Tyr Lys Asp Asp Asp Asp Lys Ile Glu Gly Arg Asn Trp

35 40 45

Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser

50 55 60

Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser

65 70 75 80

Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile

85 90 95

Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu

100 105 110

Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu

115 120 125

Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu

130 135 140

Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser

145 150 155 160

Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

165 170 175

Gly Gly Gly Ser Gly Gly Gly Ser Leu Gln Ile Thr Cys Pro Pro Pro

180 185 190

Met Ser Val Glu His Ala Asp Ile Trp Val Lys Ser Tyr Ser Leu Tyr

195 200 205

Ser Arg Glu Arg Tyr Ile Cys Asn Ser Gly Phe Lys Arg Lys Ala Gly

210 215 220

Thr Ser Ser Leu Thr Glu Cys Val Leu Asn Lys Ala Thr Asn Val Ala

225 230 235 240

His Trp Thr Thr Pro Ser Leu Lys Cys Ile Arg Asp Pro Ala Leu Val

245 250 255

His Gln Arg Pro Ala Pro Pro

260

<210> 250

<211> 117

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 250

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Ser Tyr Gly Gly Gly Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Tyr Val Asn Phe Gly Met Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 251

<211> 107

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 251

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Gly Arg Asn Pro Pro

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 252

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 252

Gly Phe Thr Phe Ser Ser Tyr Ala

15

<210> 253

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 253

Ile Gly Ser Tyr Gly Gly Gly Thr

15

<210> 254

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 254

Ala Arg Tyr Val Asn Phe Gly Met Asp Tyr

1 5 10

<210> 255

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 255

Gln Ser Ile Ser Ser Tyr

15

<210> 256

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 256

Ala Ala Ser

1

<210> 257

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 257

Gln Gln Tyr Gly Arg Asn Pro Pro Thr

15

<210> 258

<211> 116

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 258

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Val Trp Gly Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 259

<211> 107

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 259

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Gly Val Tyr Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 260

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 260

Ile Ser Gly Ser Gly Gly Ser Thr

15

<210> 261

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 261

Ala Arg Arg Val Trp Gly Phe Asp Tyr

15

<210> 262

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 262

Gln Gln Tyr Gly Val Tyr Pro Phe Thr

15

<210> 263

<211> 128

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<220>

<221> MOD_OST

<222> (120)..(120)

<223> Any amino acid

<220>

<221> MOD_OST

<222> (124)..(125)

<223> Any amino acid

<400> 263

Glu Val Arg Leu Gln Gln Ser Gly Ala Asp Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Ala Ser Gly Phe Ile Ile Lys Ala Thr Tyr

20 25 30

Met His Trp Val Arg Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile Gly

35 40 45

Arg Ile Asp Pro Ala Asn Gly Glu Lys Tyr Asp Pro Lys Phe Gln Val

50 55 60

Lys Ala Ile Thr Ala Asp Thr Ser Ser Ser Thr Ala Tyr Leu Gln Leu

65 70 75 80

Asn Ser Leu Thr Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg Tyr

85 90 95

Ala Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser

100 105 110

Ser Ala Lys Thr Thr Pro Pro Xaa Val Tyr Pro Xaa Xaa Pro Gly Ser

115 120 125

<210> 264

<211> 127

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 264

Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Thr Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Ser Phe

20 25 30

Leu Ala Trp Tyr His Gln Lys Gln Gly Arg Ser Pro Gln Leu Leu Val

35 40 45

Tyr His Ala Lys Thr Leu Ala Glu Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Gln Phe Ser Leu Lys Ile Asn Ser Leu Gln Ala

65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His Tyr Tyr Gly Ser Pro Leu

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Val Lys Arg Ala Asp Ala Ala

100 105 110

Pro Thr Val Ser Ile Phe Pro Ser Ser Glu Glu Leu Ser Leu

115 120 125

<210> 265

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 265

Gly Phe Ile Ile Lys Ala Thr Tyr Met His

1 5 10

<210> 266

<211> 17

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 266

Arg Ile Asp Pro Ala Asn Gly Glu Thr Lys Tyr Asp Pro Lys Phe Gln

1 5 10 15

Val

<210> 267

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 267

Tyr Ala Trp Tyr Phe Asp Val

15

<210> 268

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 268

Arg Ala Ser Glu Asn Ile Tyr Ser Phe Leu Ala

1 5 10

<210> 269

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 269

His Ala Lys Thr Leu Ala Glu

15

<210> 270

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 270

Gln His Tyr Tyr Gly Ser Pro Leu Thr

15

<210> 271

<211> 127

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 271

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Trp Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Ala Ile Asn Gln Asp Gly Ser Glu Lys Tyr Tyr Val Gly Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Val Arg Asp Tyr Tyr Asp Ile Leu Thr Asp Tyr Tyr Ile His Tyr Trp

100 105 110

Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 272

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 272

Asn Tyr Trp Met Asn

15

<210> 273

<211> 17

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 273

Ala Ile Asn Gln Asp Gly Ser Glu Lys Tyr Tyr Val Gly Ser Val Lys

1 5 10 15

gly

<210> 274

<211> 18

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 274

Asp Tyr Tyr Asp Ile Leu Thr Asp Tyr Tyr Ile His Tyr Trp Tyr Phe

1 5 10 15

Asp Leu

<210> 275

<211> 12

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 275

Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala

1 5 10

<210> 276

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 276

Gly Ala Ser Ser Arg Ala Thr

15

<210> 277

<211> 446

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 277

Glu Val Gln Leu Val Glu Ser Gly Gly Asp Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Arg Gly Ser Leu Tyr Tyr Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 278

<211> 214

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 278

Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Pro Ser Gln Gly Ile Asn Trp Glu

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Glu Gln Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 279

<211> 119

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 279

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr

20 25 30

His Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Val Ile Asn Pro Met Tyr Gly Thr Thr Asp Tyr Asn Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Tyr Asp Tyr Phe Thr Gly Thr Gly Val Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 280

<211> 112

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 280

Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Arg Ser Leu Val His Ser

20 25 30

Arg Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ile Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ser Gln Ser

85 90 95

Thr His Leu Pro Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 281

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 281

Arg Tyr Gly Ile Ser

15

<210> 282

<211> 17

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 282

Trp Ile Ser Thr Tyr Ser Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln

1 5 10 15

gly

<210> 283

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 283

Arg Gln Leu Tyr Phe Asp Tyr

15

<210> 284

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 284

Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala

1 5 10

<210> 285

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 285

Asp Ala Ser Thr Arg Ala Thr

15

<210> 286

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 286

Gln Gln Tyr Asp Asn Trp Pro Leu Thr

15

<210> 287

<211> 137

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 287

Met Glu Phe Gly Leu Ser Trp Val Phe Leu Val Ala Leu Leu Arg Gly

1 5 10 15

Val Gln Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln

20 25 30

Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe

35 40 45

Ser Val Tyr Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu

50 55 60

Glu Trp Val Ala Ile Ile Trp Tyr Asp Gly Asp Asn Gln Tyr Tyr Ala

65 70 75 80

Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn

85 90 95

Thr Leu Tyr Leu Gln Met Asn Gly Leu Arg Ala Glu Asp Thr Ala Val

100 105 110

Tyr Tyr Cys Ala Arg Asp Leu Arg Thr Gly Pro Phe Asp Tyr Trp Gly

115 120 125

Gln Gly Thr Leu Val Thr Val Ser Ser

130 135

<210> 288

<211> 126

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 288

Met Leu Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala

1 5 10 15

Ser Arg Gly Glu Ile Val Leu Thr Gln Ser Pro Asp Phe Gln Ser Val

20 25 30

Thr Pro Lys Glu Lys Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile

35 40 45

Gly Ser Ser Leu His Trp Tyr Gln Gln Lys Pro Asp Gln Ser Pro Lys

50 55 60

Leu Leu Ile Lys Tyr Ala Ser Gln Ser Phe Ser Gly Val Pro Ser Arg

65 70 75 80

Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser

85 90 95

Leu Glu Ala Glu Asp Ala Ala Ala Tyr Tyr Cys His Gln Ser Ser Ser

100 105 110

Leu Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

115 120 125

<210> 289

<211> 12

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 289

Gly Asp Ser Val Ser Ser Asn Ser Ala Ala Trp Gly

1 5 10

<210> 290

<211> 18

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 290

Arg Ile Tyr Tyr Arg Ser Lys Trp Tyr Asn Ser Tyr Ala Val Ser Val

1 5 10 15

Lys Ser

<210> 291

<211> 12

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 291

Arg Ala Ser Gln Phe Ile Ser Ser Tyr Leu Ser

1 5 10

<210> 292

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 292

Leu Leu Ile Tyr Gly Ser Ser Ser Arg Ala Thr

1 5 10

<210> 293

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 293

Gln Gln Leu Tyr Ser Ser Pro Met

15

<210> 294

<211> 124

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 294

Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn

20 25 30

Ser Ala Ala Trp Gly Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Ile Tyr Tyr Arg Ser Lys Trp Tyr Asn Ser Tyr Ala

50 55 60

Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Tyr Asp Trp Val Pro Lys Ile Gly Val Phe Asp

100 105 110

Ser Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 295

<211> 110

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 295

Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Phe Ile Ser Ser Ser

20 25 30

Tyr Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ser Ser Ser Arg Ala Thr Gly Val Pro Ala Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Leu Tyr Ser Ser Pro

85 90 95

Met Thr Phe Gly Glyn Gly Thr Lys Val Glu Ile Lys Arg Thr

100 105 110

<210> 296

<211> 454

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 296

Gln Val Gln Leu Gln Gln Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Ile Ser Gly Asp Ser Val Ser Ser Asn

20 25 30

Ser Ala Ala Trp Gly Trp Ile Arg Gln Ser Pro Gly Arg Gly Leu Glu

35 40 45

Trp Leu Gly Arg Ile Tyr Tyr Arg Ser Lys Trp Tyr Asn Ser Tyr Ala

50 55 60

Val Ser Val Lys Ser Arg Ile Thr Ile Asn Pro Asp Thr Ser Lys Asn

65 70 75 80

Gln Phe Ser Leu Gln Leu Asn Ser Val Thr Pro Glu Asp Thr Ala Val

85 90 95

Tyr Tyr Cys Ala Arg Tyr Asp Trp Val Pro Lys Ile Gly Val Phe Asp

100 105 110

Ser Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys

115 120 125

Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly

130 135 140

Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro

145 150 155 160

Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

165 170 175

Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val

180 185 190

Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn

195 200 205

Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro

210 215 220

Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu

225 230 235 240

Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

245 250 255

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

260 265 270

Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly

275 280 285

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn

290 295 300

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

305 310 315 320

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro

325 330 335

Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

340 345 350

Pro Gln Val Tyr Thr Leu Pro Ser Arg Glu Glu Met Thr Lys Asn

355 360 365

Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

370 375 380

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

385 390 395 400

Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

405 410 415

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

420 425 430

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

435 440 445

Ser Leu Ser Pro Gly Lys

450

<210> 297

<211> 215

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 297

Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Phe Ile Ser Ser Ser

20 25 30

Tyr Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ser Ser Ser Arg Ala Thr Gly Val Pro Ala Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Leu Tyr Ser Ser Pro

85 90 95

Met Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 298

<211> 109

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 298

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Cys Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg

100 105

<210> 299

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 299

Gln Gln Tyr Gly Ser Ser Pro Cys Thr

15

<210> 300

<211> 12

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 300

Tyr Asp Trp Val Pro Lys Ile Gly Val Phe Asp Ser

1 5 10

<210> 301

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 301

Gly Phe Ser Leu Ser Ser Tyr

15

<210> 302

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 302

Trp Gly Gly Gly Gly

15

<210> 303

<211> 13

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 303

His Ala Tyr Gly His Asp Gly Gly Phe Ala Met Asp Tyr

1 5 10

<210> 304

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 304

Ser Glu Ser Val Ser Ser Asn

15

<210> 305

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 305

Gly Ala Ser

1

<210> 306

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 306

Ser Tyr Ser Tyr Pro Phe

15

<210> 307

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 307

Ser Tyr Gly Val Asp

15

<210> 308

<211> 16

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 308

Val Ile Trp Gly Gly Gly Gly Thr Tyr Tyr Ala Ser Ser Leu Met Gly

1 5 10 15

<210> 309

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 309

Arg Ala Ser Glu Ser Val Ser Ser Asn Val Ala

1 5 10

<210> 310

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 310

Gly Ala Ser Asn Arg Ala Thr

15

<210> 311

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

peptide"

<400> 311

Gly Gln Ser Tyr Ser Tyr Pro Phe Thr

15

<210> 312

<211> 121

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 312

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ser Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Leu Ser Ser Tyr

20 25 30

Gly Val Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Glu Trp Val

35 40 45

Gly Val Ile Trp Gly Gly Gly Gly Thr Tyr Tyr Ala Ser Ser Leu Met

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg His Ala Tyr Gly His Asp Gly Gly Phe Ala Met Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 313

<211> 107

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 313

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Ser Asn

20 25 30

Val Ala Trp Tyr Gln Gln Arg Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gly Gln Ser Tyr Ser Tyr Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 314

<211> 451

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 314

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ser Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Leu Ser Ser Tyr

20 25 30

Gly Val Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Glu Trp Val

35 40 45

Gly Val Ile Trp Gly Gly Gly Gly Thr Tyr Tyr Ala Ser Ser Leu Met

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg His Ala Tyr Gly His Asp Gly Gly Phe Ala Met Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala

130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

165 170 175

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

180 185 190

Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His

195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys

210 215 220

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

225 230 235 240

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

245 250 255

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His

260 265 270

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

290 295 300

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

305 310 315 320

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile

325 330 335

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

340 345 350

Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly Lys

450

<210> 315

<211> 214

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 315

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Glu Ser Val Ser Ser Asn

20 25 30

Val Ala Trp Tyr Gln Gln Arg Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gly Gln Ser Tyr Ser Tyr Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 316

<211> 120

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 316

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met Tyr Trp Val Arg Gln Ala Arg Gly Gln Arg Leu Glu Trp Ile

35 40 45

Gly Arg Ile Asp Pro Asn Ser Gly Ser Thr Lys Tyr Asn Glu Lys Phe

50 55 60

Lys Asn Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Tyr Arg Lys Gly Leu Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 317

<211> 107

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 317

Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala

20 25 30

Val Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile

35 40 45

Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Glu Ala

65 70 75 80

Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 318

<211> 120

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 318

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met Tyr Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Asp Pro Asn Ser Gly Ser Thr Lys Tyr Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Tyr Arg Lys Gly Leu Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 319

<211> 107

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 319

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ala Ser Gln Asp Val Gly Thr Ala

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Leu

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 320

<211> 125

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 320

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Leu Thr Asn Tyr

20 25 30

Gly Met Asn Trp Val Arg Gln Ala Arg Gly Gln Arg Leu Glu Trp Ile

35 40 45

Gly Trp Ile Asn Thr Asp Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asn Pro Pro Tyr Tyr Tyr Gly Thr Asn Asn Ala Glu Ala Met

100 105 110

Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120 125

<210> 321

<211> 107

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 321

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ser Ser Gln Asp Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Thr Leu His Leu Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Asn Leu Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 322

<211> 125

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 322

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Leu Thr Asn Tyr

20 25 30

Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Asp Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asn Pro Pro Tyr Tyr Tyr Gly Thr Asn Asn Ala Glu Ala Met

100 105 110

Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120 125

<210> 323

<211> 107

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 323

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ser Ser Gln Asp Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Thr Leu His Leu Gly Ile Pro Arg Phe Ser Gly

50 55 60

Ser Gly Tyr Gly Thr Asp Phe Thr Leu Thr Ile Asn Asn Ile Glu Ser

65 70 75 80

Glu Asp Ala Ala Tyr Tyr Phe Cys Gln Gln Tyr Tyr Asn Leu Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 324

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 324

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Asp Ile Tyr Pro Gly Gln Gly Asp Thr Ser Tyr Asn Gln Lys Phe

50 55 60

Lys Gly Arg Ala Thr Met Thr Ala Asp Lys Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Gly Gly Ala Phe Pro Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 325

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 325

Asp Ile Val Leu Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Glu Ser Val Glu Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Asp

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln Ser Arg

85 90 95

Lys Asp Pro Ser Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 326

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 326

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Asn Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Asp Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Gly Gly Ala Phe Pro Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Thr Val Thr Val Ser Ser

115

<210> 327

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Artificial Sequence Description: Synthetic

polypeptide"

<400> 327

Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Ser Val Glu Tyr Tyr

20 25 30

Gly Thr Ser Leu Met Gln Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Ala Ala Ser Asn Val Glu Ser Gly Val Pro Ser

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ser Arg

85 90 95

Lys Asp Pro Ser Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<---

Claims (172)

1. Use of an anti-PD-1 antibody molecule at a dose of approximately 300 mg to 400 mg once every three weeks or once every four weeks in the treatment of cancer in a subject, where the anti-PD-1 antibody molecule comprises: (a) a heavy chain variable region (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 light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 13, the VLCDR2 amino acid sequence of SEQ ID NO: 14, and the VLCDR3 amino acid sequence of SEQ ID NO: 33; (b) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 1; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32; (c) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224, the amino acid sequence of VHCDR2 SEQ ID NO: 5, and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33; or (d) VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32. 2. A method of treating cancer in a subject, comprising administering to the subject an anti-PD-1 antibody molecule at a dose of about 300 mg to 400 mg once every three weeks or once every four weeks, where the anti-PD-1 antibody molecule comprises: (a) a heavy chain variable region (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 light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 13, the VLCDR2 amino acid sequence of SEQ ID NO: 14, and the VLCDR3 amino acid sequence of SEQ ID NO: 33; (b) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 1; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32; (c) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224, the amino acid sequence of VHCDR2 SEQ ID NO: 5, and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33; or (d) VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32. 3. The use of claim 1 or the method of claim 2, wherein the anti-PD-1 antibody molecule is used in combination with one or more therapeutic agents selected from one, two or all of the following categories (i)-(iii): (i) an agent that increases tumor antigen presentation, selected from one or more of the following: STING agonist (interferon response stimulator cGAMP interactor 1), TLR (Toll-like receptor) agonist, A2AR (adenosine A2a receptor) antagonist, oncolytic virus, TIM-3 modulator (T-cell immunoglobulin and mucin-3 domain T-cell immunoglobulin and mucin-containing domain-3), vascular endothelial growth factor receptor (VEGFR) inhibitor, c-Met (tyrosine protein kinase Met) inhibitor, TGFb inhibitor ( transforming growth factor beta), IDO/TDO inhibitor (indolamine-2,3-dioxygenase/tryptophan-2,3-dioxygenase), vaccine or bi- or trispecific cell activator; (ii) an agent that enhances the response of effector cells, selected from one or more of the following: GITR (glucocorticoid-induced TNFR-related protein) agonist, PD-L1 (programmed death protein ligand 1) inhibitor, IAP (apoptosis protein inhibitor), inhibitor of EGFR (epidermal growth factor receptor), inhibitor of the target of rapamycin (mTOR), IL-15 (interleukin-15) or a variant thereof, inhibitor of CTLA-4 (cytotoxic T-lymphocyte-associated protein 4), a bispecific antibody molecule that binds to CD3 (cluster of differentiation 3) and a tumor antigen, CD40 agonist (cluster of differentiation 40), agonist of OX40 (a member of the tumor necrosis factor receptor superfamily 4) or CD27 agonist (cluster of differentiation 27); or (iii) an agent that reduces tumor immunosuppression, selected from one or more of the following: a GITR agonist, an inhibitor of an immune checkpoint molecule selected from one or more of PD-L1, LAG-3 (lymphocyte activation gene 3), TIM-3, or CTLA-4, CSF-1/1R (colony stimulating factor-1/colony stimulating factor-1 receptor) inhibitor, IL-17 (interleukin-17) inhibitor, IL-1β (interleukin-1 beta) inhibitor, CXCR2 (chemokine receptor motif) inhibitor C-X-C 2), PI3Kγ (phosphoinositide-3-kinase gamma) or PI3Kδ (phosphoinositide-3-kinase delta) inhibitor, BAFF-R (B-cell activating factor receptor) inhibitor, MALT-1/BTK (lymphoid tissue translocation protein 1) inhibitor mucosal-associated/Bruton's tyrosine kinase), JAK (Janus kinase) inhibitor, CRTH2 (prostaglandin D2 receptor 2) inhibitor, VEGFR (vascular endothelial growth factor receptor), IL-15 or variant, CTLA-4 inhibitor, IDO inhibitor /TDO, A2AR antagonist, inhibitor TGFb or PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3) inhibitor. 4. The use of claim 1 or 3, or the method of claim 2 or 3, wherein the anti-PD-1 antibody molecule is applied at a dose of approximately 300 mg once every three weeks. 5. The use of claim 1 or 3, or the method of claim 2 or 3, wherein the anti-PD-1 antibody molecule is applied at a dose of approximately 400 mg once every four weeks. 6. Application according to any one of paragraphs. 1 or 3-5 or the method according to any one of paragraphs. 2-5, wherein the anti-PD-1 antibody molecule comprises: (a) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 42; (b) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 66; (d) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 70; (d) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 70; (e) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 46; (f) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 46; (g) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 54; (h) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 54; (i) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 58; (j) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 62; (k) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 66; (l) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 74; (m) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 78; (n) VH comprising the amino acid sequence of SEQ ID NO: 82 and VL comprising the amino acid sequence of SEQ ID NO: 70; (o) VH comprising the amino acid sequence of SEQ ID NO: 82 and VL comprising the amino acid sequence of SEQ ID NO: 66; or (p) VH comprising the amino acid sequence of SEQ ID NO: 86 and VL comprising the amino acid sequence of SEQ ID NO: 66. 7. Application according to any one of paragraphs. 1 or 3-6 or the method according to any one of paragraphs. 2-6, where the cancer is a solid tumor or hemoblastosis. 8. Application according to any one of paragraphs. 1 or 3-7 or the method according to any one of paragraphs. 2-7, where the cancer is selected from lung cancer, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, colon and rectal cancer (CRC), gastric cancer, pancreatic cancer, thyroid cancer, neuroendocrine tumor ( NET), ovarian cancer, fallopian tube cancer, lymphoma, leukemia, myelodysplastic syndrome (MDS), or metastatic cancer. 9. Use according to claim 8 or method according to claim 8, wherein the skin cancer is Merkel cell cancer or melanoma. 10. The use of claim 8 or the method of claim 8 wherein the breast cancer is triple negative breast cancer (TNBC), HER2 negative breast cancer, or ER+ breast cancer. 11. Use according to claim 8 or method according to claim 8, wherein the kidney cancer is renal cell carcinoma. 12. The use of claim 11 or the method of claim 11 wherein the renal cell carcinoma is clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC). 13. Use according to claim 8 or method according to claim 8, wherein the thyroid cancer is anaplastic thyroid cancer (ATC). 14. The use of claim 8 or the method of claim 8 wherein the neuroendocrine tumor (NET) is an atypical pulmonary carcinoid tumor or NET in the pancreas, gastrointestinal (GI) tract, or lung. 15. Use according to claim 8 or method according to claim 8, wherein the lung cancer is non-small cell lung cancer (NSCLC). 16. The use of claim 15 or the method of claim 15, wherein the NSCLC is squamous NSCLC, non-squamous NSCLC, or NSCLC adenocarcinoma. 17. Use according to claim 8 or method according to claim 8, wherein the colon and rectal cancer (CRC) is highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC) ). 18. Use according to claim 8 or method according to claim 8, wherein the leukemia is acute myeloid leukemia (AML). 19. The use of claim 18 or the method of claim 18 wherein the acute myeloid leukemia (AML) is recurrent or refractory AML or de novo AML. 20. The use of claim 8 or the method of claim 8 wherein the myelodysplastic syndrome (MDS) is a high risk MDS. 21. Use according to claim 8 or method according to claim 8, wherein the liver cancer is hepatocellular carcinoma. 22. Use according to claim 8 or method according to claim 8, wherein the gastric cancer is EBV+ gastric cancer. 23. Use according to claim 8 or method according to claim 8, wherein the lymphoma is non-Hodgkin's lymphoma. 24. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with an IAP inhibitor. 25. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with an mTOR inhibitor. 26. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with a DAC inhibitor. 27. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with an IL-1β inhibitor. 28. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with an IL-17 inhibitor. 29. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with a MEK inhibitor. 30. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with an EGFR inhibitor. 31. Application according to any one of paragraphs. 24-30 or the method according to any one of paragraphs. 24-30, where the combination is used to treat colon and rectal cancer, lung cancer or breast cancer. 32. Use according to claim 31 or method according to claim 31, wherein the lung cancer is NSCLC. 33. Use according to claim 31 or method according to claim 31, wherein the breast cancer is TNBC. 34. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with a CSF-1/1R binding agent. 35. The use of claim 34, or the method of claim 34, wherein the combination is used to treat brain cancer, breast cancer, or pancreatic cancer. 36. The use of claim 35 or the method of claim 35 wherein the brain cancer is glioblastoma multiforme (GBM). 37. Use according to claim 35 or method according to claim 35, wherein the breast cancer is TNBC. 38. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with a 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) inhibitor. 39. The use of claim 38, or the method of claim 38, wherein the combination is used to treat ovarian cancer, fallopian tube cancer, or colorectal cancer (CRC). 40. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is used in combination with a chemotherapeutic agent, where optionally the chemotherapeutic agent is paclitaxel. 41. The use of claim 40 or the method of claim 40 wherein the combination is used to treat breast cancer. 42. The use of claim 41 or the method of claim 41, wherein the breast cancer is HER2-negative breast cancer. 43. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is administered in combination with a BRAF inhibitor and a MEK inhibitor. 44. Application according to paragraph 43 or method according to paragraph 43, wherein the anti-PD-1 antibody is administered at a dose of approximately 400 mg once every four weeks, wherein the BRAF inhibitor is used at a dose of 50 mg to 250 mg twice daily, and where the MEK inhibitor is used at a dose of 1 mg to 3 mg once a day. 45. The use of claim 44 or the method of claim 44, wherein the BRAF inhibitor is used at a dose of about 150 mg twice daily and/or the MEK inhibitor is used at a dose of about 2 mg once daily. 46. The use of claim 44 or 45, or the method of claim 44 or 45, wherein the anti-PD-1 antibody molecule is administered intravenously and/or where the BRAF inhibitor and/or MEK inhibitor is administered orally. 47. Application according to any one of paragraphs. 44-46 or the method according to any one of paragraphs. 44-46 wherein the BRAF inhibitor is dabrafenib and/or where the MEK inhibitor is trametinib. 48. Application according to any one of paragraphs. 44-47 or the method according to any one of paragraphs. 44-47, which is for the treatment of melanoma, optionally, where the melanoma has a BRAF mutation, optionally, where the BRAF mutation is a BRAF V600 mutation. 49. Application according to any one of paragraphs. 43-47 or the method according to any one of paragraphs. 43-47, where the combination is used to treat skin cancer, lung cancer, colon and rectal cancer, or pancreatic cancer. 50. The use of claim 49 or the method of claim 49 wherein the skin cancer is melanoma or where the lung cancer is non-small cell lung cancer. 51. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is administered in combination with a TGF-β inhibitor. 52. The use of claim 51 or the method of claim 51 wherein the combination is used to treat pancreatic cancer, colon and rectal cancer, lung cancer, breast cancer, liver cancer, prostate cancer, or kidney cancer. 53. The use of claim 52 or the method of claim 52 wherein the colon and rectal cancer is microsatellite stable colon and rectal cancer (MSS-CRC). 54. Use according to claim 52 or method according to claim 52, wherein the lung cancer is NSCLC. 55. Use according to claim 52 or method according to claim 52, wherein the breast cancer is TNBC. 56. The use of claim 52 or the method of claim 52 wherein the liver cancer is hepatocellular carcinoma. 57. The use of claim 52, or the method of claim 52, wherein the kidney cancer is clear cell renal cell carcinoma. 58. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is administered in combination with a PD-L1 inhibitor. 59. The use of claim 58, or the method of claim 58, wherein the combination is used to treat lung cancer, breast cancer, uterine cancer, or thyroid cancer. 60. Use according to claim 59 or method according to claim 59, wherein the lung cancer is NSCLC. 61. Use according to claim 59 or method according to claim 59, wherein the breast cancer is TNBC. 62. The use of claim 59, or the method of claim 59, wherein the uterine cancer is endometrial carcinoma. 63. The use of claim 59 or the method of claim 59, wherein the thyroid cancer is an anaplastic thyroid carcinoma. 64. Application according to any one of paragraphs. 1 or 3-23 or the method according to any one of paragraphs. 2-23 wherein the anti-PD-1 antibody molecule is administered in combination with a hypomethylating agent. 65. The use of claim 64 or the method of claim 64, wherein the combination further comprises a TIM-3 inhibitor. 66. The use of claim 64 or 65, or the method of claim 64 or 65, wherein the combination is used to treat leukemia or myelodysplastic syndrome (MDS). 67. The use of claim 66 or the method of claim 66 wherein the leukemia is acute myeloid leukemia (AML). 68. The use of claim 66 or the method of claim 66 wherein the MDS is high risk MDS. 69. Application according to any one of paragraphs. 1 or 3-68 or the method according to any one of paragraphs. 2-68, wherein the anti-PD-1 antibody molecule is used in combination with a second PD-1 inhibitor. 70. The use of a pharmaceutical composition comprising an anti-PD-1 antibody molecule at a dose of approximately 300 mg to 400 mg once every three weeks or once every four weeks for the treatment of cancer, where the anti-PD-1 antibody molecule comprises: (a) a heavy chain variable region (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 light chain variable region (VL) comprising the VLCDR1 amino acid sequence of SEQ ID NO: 13, the VLCDR2 amino acid sequence of SEQ ID NO: 14, and the VLCDR3 amino acid sequence of SEQ ID NO: 33; (b) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 1; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32; (c) a VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224, the amino acid sequence of VHCDR2 SEQ ID NO: 5, and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 13, the amino acid sequence of VLCDR2 SEQ ID NO: 14, and the amino acid sequence of VLCDR3 SEQ ID NO: 33; or (d) VH comprising the amino acid sequence of VHCDR1 SEQ ID NO: 224; the amino acid sequence of VHCDR2 SEQ ID NO: 2 and the amino acid sequence of VHCDR3 SEQ ID NO: 3; and a VL comprising the amino acid sequence of VLCDR1 SEQ ID NO: 10, the amino acid sequence of VLCDR2 SEQ ID NO: 11, and the amino acid sequence of VLCDR3 SEQ ID NO: 32. 71. Use according to claim 70, where the dose is approximately 300 mg once every three weeks. 72. Use according to claim 70, where the dose is approximately 400 mg once every four weeks. 73. Application according to any one of paragraphs. 70-72, wherein the anti-PD-1 antibody molecule comprises: (a) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 42; (b) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 66; (d) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 70; (d) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 70; (e) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 46; (f) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 46; (g) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 54; (h) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 54; (i) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 58; (j) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 62; (k) VH comprising the amino acid sequence of SEQ ID NO: 50 and VL comprising the amino acid sequence of SEQ ID NO: 66; (l) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 74; (m) VH comprising the amino acid sequence of SEQ ID NO: 38 and VL comprising the amino acid sequence of SEQ ID NO: 78; (n) VH comprising the amino acid sequence of SEQ ID NO: 82 and VL comprising the amino acid sequence of SEQ ID NO: 70; (o) VH comprising the amino acid sequence of SEQ ID NO: 82 and VL comprising the amino acid sequence of SEQ ID NO: 66; or (p) VH comprising the amino acid sequence of SEQ ID NO: 86 and VL comprising the amino acid sequence of SEQ ID NO: 66. 74. Application according to any one of paragraphs. 70-73, where the cancer is selected from lung cancer, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, colon and rectal cancer, gastric cancer, pancreatic cancer, thyroid cancer, neuroendocrine tumor (NET), ovarian cancer, fallopian tube cancer, lymphoma, leukemia, myelodysplastic syndrome (MDS), or metastatic cancer. 75. Application according to any one of paragraphs. 70-73, where the skin cancer is Merkel cell cancer or melanoma. 76. The use of claim 74 wherein the breast cancer is triple negative breast cancer (TNBC), HER2 negative breast cancer, ER+ breast cancer. 77. Use according to claim 74, wherein the kidney cancer is renal cell carcinoma. 78. The use of claim 77 wherein the renal cell carcinoma is clear cell renal cell carcinoma (CCRCC) or non-clear cell renal cell carcinoma (nccRCC). 79. The use of claim 74 wherein the thyroid cancer is anaplastic thyroid carcinoma (ATC). 80. The use of claim 74 wherein the neuroendocrine tumor (NET) is an atypical pulmonary carcinoid tumor or NET in the pancreas, gastrointestinal (GI) tract, or lung. 81. The use of claim 74 wherein the lung cancer is non-small cell lung cancer (NSCLC). 82. The use of claim 81, wherein the NSCLC is squamous NSCLC, non-squamous NSCLC, or NSCLC adenocarcinoma. 83. Use according to claim 74, wherein the colon and rectal cancer (CRC) is highly microsatellite unstable colon and rectal cancer (MSI-H CRC) or microsatellite stable colon and rectal cancer (MSS CRC). 84. The use of claim 74 wherein the leukemia is acute myeloid leukemia (AML). 85. Use according to claim 84, wherein acute myeloid leukemia (AML) is recurrent or refractory AML, or de novo AML. 86. The use of claim 74 wherein the myelodysplastic syndrome (MDS) is a high risk MDS. 87. Use of an anti-PD-1 antibody molecule at a dose of approximately 300 mg once every three weeks or approximately 400 mg once every four weeks in the treatment of cancer in a subject where the anti-PD-1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 38 and a VL comprising the amino acid sequence of SEQ ID NO: 54. 88. Use of an anti-PD-1 antibody molecule at a dose of approximately 300 mg once every three weeks or approximately 400 mg once every four weeks in the treatment of cancer in a subject where the anti-PD-1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 38 and a VL comprising the amino acid sequence of SEQ ID NO: 70. 89. Use of an anti-PD-1 antibody molecule at a dose of approximately 300 mg once every three weeks or approximately 400 mg once every four weeks in the treatment of cancer in a subject wherein the anti-PD-1 antibody molecule 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: 56. 90. Use of an anti-PD-1 antibody molecule at a dose of about 300 mg once every three weeks or about 400 mg once every four weeks in the treatment of cancer in a subject, wherein the anti-PD-1 antibody molecule 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: 72. 91. A method of treating cancer in a subject, comprising administering to the subject an anti-PD-1 antibody molecule at a dose of about 300 mg once every three weeks or about 400 mg once every four weeks, where the anti-PD-1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 38 and a VL comprising the amino acid sequence of SEQ ID NO: 54. 92. A method of treating cancer in a subject, comprising administering to the subject an anti-PD-1 antibody molecule at a dose of about 300 mg once every three weeks or about 400 mg once every four weeks, where the anti-PD-1 antibody molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 38 and a VL comprising the amino acid sequence of SEQ ID NO: 70. 93. A method of treating cancer in a subject, comprising administering to the subject an anti-PD-1 antibody molecule at a dose of about 300 mg once every three weeks or about 400 mg once every four weeks, wherein the anti-PD-1 antibody molecule 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: 56. 94. A method of treating cancer in a subject, comprising administering to the subject an anti-PD-1 antibody molecule at a dose of about 300 mg once every three weeks or about 400 mg once every four weeks, wherein the anti-PD-1 antibody molecule 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: 72. 95. Use of an anti-PD-1 antibody molecule at a dose of 300 mg once every three weeks or 400 mg once every four weeks in the treatment of cancer in a subject where: the anti-PD-1 antibody molecule comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 91 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-PD-1 antibody molecule is used in combination with dabrafenib and trametinib. 96. Use of an anti-PD-1 antibody molecule at a dose of 300 mg once every three weeks or 400 mg once every four weeks in the treatment of cancer in a subject where: the anti-PD-1 antibody molecule comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 91 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 72; and where the anti-PD-1 antibody molecule is used in combination with dabrafenib, where dabrafenib is used at a dose of 150 mg twice a day, and in combination with trametinib, where trametinib is used at a dose of 2 mg once a day. 97. A method of treating cancer in a subject, comprising administering to the subject an anti-PD-1 antibody molecule at a dose of 300 mg once every three weeks or 400 mg once every four weeks, where: the anti-PD-1 antibody molecule comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 91 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-PD-1 antibody molecule is used in combination with dabrafenib and trametinib. 98. A method of treating cancer in a subject, comprising administering to the subject an anti-PD-1 antibody molecule at a dose of 300 mg once every three weeks or 400 mg once every four weeks, where: the anti-PD-1 antibody molecule comprises a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 91 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 72; and where the anti-PD-1 antibody molecule is used in combination with dabrafenib, where dabrafenib is used at a dose of 150 mg twice a day, and in combination with trametinib, where trametinib is used at a dose of 2 mg once a day. 99. Application according to any one of paragraphs. 85-90, 95 or 96 or the method according to any one of paragraphs. 91-94, 97 or 98, where the cancer includes a BRAF mutation. 100. The use of claim 99 or the method of claim 99, wherein the cancer is a BRAF V600E mutant melanoma. 101. The use of claim 99 or the method of claim 99, wherein the cancer is a BRAF V600E mutant non-small cell lung cancer. 102. The use of claim 99 or the method of claim 99, wherein the cancer is BRAF V600E colon and rectal cancer. 103. The use of claim 99 or the method of claim 99, wherein the cancer is BRAF V600E pancreatic cancer.

Images