Drug combinations of anti-CEACAM6 and anti-PD-1 or anti-PD-L1 antibodies for the treatment of cancer

Abstract

Drug combination for treating cancer The present invention relates to a combination of at least two components (component A and component B) for treating cancer, component A is an anti-CEACAM6 antibody, component B is an anti-PD-1 antibody or an anti-PD-L1 antibody, the anti-PD-1 antibody is preferably nivolumab or pembrolizumab, and the anti-PD-L1 antibody is preferably atezolizumab, avelumab or durvalumab.

Description

[0001] The present invention relates to a combination of at least two components (component A and component B), component A is the anti-CEACAM6 antibody TPP-3310, and component B is an anti-PD-1 antibody or an anti-PD-L1 antibody, the anti-PD-1 antibody is preferably nivolumab or pembrolizumab, and the anti-PD-L1 antibody is preferably atezolizumab, avelumab or durvalumab.

[0002] Another aspect of the invention relates to the use of such combinations as described herein for the preparation of a medicament for the treatment or prevention of cancer.

[0003] Yet another aspect of the invention relates to a method of treating or preventing cancer in a subject, comprising administering to the subject a therapeutically effective amount of a combination as described herein.

[0004] Furthermore, the present invention relates to a kit comprising the following combination:

[0005] - Component A, which is the anti-CEACAM6 antibody TPP-3310;

[0006] - component B, which is an anti-PD-1 antibody or an anti-PD-L1 antibody, the anti-PD-1 antibody is preferably nivolumab or pembrolizumab, the anti-PD-L1 antibody is preferably atezolizumab, avelumab or durvalumab, and optionally

[0007] - one or more pharmaceutical preparations C;

[0008] Optionally, one or both of the components A and B are in the form of ready-for-use pharmaceutical preparations for simultaneous, concurrent, separate or sequential administration.

[0009] Components A and B are preferably administered by the intravenous route.

[0010] In some embodiments, the cancer is lung cancer, in particular non-small cell lung cancer (NSCLC), pancreatic cancer, gastric cancer, colorectal cancer, head and neck cancer, bladder cancer, bile duct cancer, breast cancer, cervical cancer, esophageal cancer. Background Art

[0011] Cancer is the second most common cause of death in the United States, accounting for 450,000 deaths each year. Although substantial progress has been made in identifying some of the possible environmental and genetic causes of cancer, there remains a need for additional therapeutic modalities targeting cancer and related diseases. In particular, there is a need for therapeutic approaches to treat diseases associated with dysregulated growth / proliferation.

[0012] Cancer is a complex disease resulting from a selection process for cells with acquired functional capabilities such as enhanced viability / resistance to apoptosis and unlimited proliferative potential. Therefore, it is preferred to develop drugs for cancer therapy that address the different features of established tumors.

[0013] T cell responses to tumor-associated antigens have been described in many tumors, which usually cause the accumulation of tumor-specific memory T cells in lymphoid organs or in the blood. However, the ability of T cells to respond to autologous tumor cells is generally low. Many tumors have the ability to block the effector function of T cells, which leads to limited activity of tumor immunotherapy. For a variety of cancers, the unresponsiveness of T cells to tumor cells has been confirmed.

[0014] The immune system relies on multiple checkpoints, or "immunological brakes," to prevent the immune system from becoming overly activated on healthy cells. Tumor cells often exploit these checkpoints to evade detection by the immune system. CTLA-4 and PD-1 are checkpoints that have been studied as targets for cancer therapy.

[0015] Checkpoint proteins regulate T cell activation or function. Many checkpoint proteins are known, such as CTLA-4 and its ligands CD80 and CD86; and PD-1 and its ligands PD-L1 and PD-L2. These proteins are responsible for the co-stimulatory or inhibitory interactions of T cell responses. Immune checkpoint proteins regulate and maintain self-tolerance and the duration and amplitude of physiological immune responses. Immune checkpoint inhibitors include antibodies or are derived from antibodies. Currently, different immunotherapy approaches are becoming powerful treatment strategies for various malignant diseases.

[0016] A very famous recent example of successful cancer immunotherapy involves immune checkpoint blockade therapy, which is achieved by targeting inhibitory molecules on immune effector T cells or antigen presenting cells (including tumor cells) with monoclonal antibodies (mAbs). Interference with co-inhibitors has been shown to unleash powerful anti-tumor T cell responses (Pardoll: The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 12: (2012) 252–264).

[0017] CTLA-4 has been shown to be abnormally upregulated in certain cancers and present on the surface of T cells, thereby inhibiting T cell activation in response to tumor cells. PD-1 is another immunological checkpoint that has been found to be upregulated in certain tumors; it inhibits T cell function, thereby contributing to the tumor's ability to evade the immune system.

[0018] Antibody blockade of immune checkpoint molecules for immune cell activation and thus for cancer immunotherapy is a clinically validated method. In 2011, the CTLA-4 blocking antibody Ipilimumab was approved by the FDA for second-line therapy of metastatic melanoma (Yervoy). Another example is blocking the PD-1 / PD-L1 axis, for which several drugs have been approved or are being clinically developed, and impressive clinical responses have been reported in melanoma, lung cancer, RCC, bladder cancer, etc. (Shen and Zhao: Efficacy of PD-1 and PD-L1 inhibitors and PD-L1 expression status in cancer: meta-analysis. BMJ2018; 362: k3529).

[0019] In 2013, it was reported that combination therapy of anti-CTLA4 and anti-PD1 mAbs could synergistically improve survival and tumor regression in patients with advanced melanoma (Wolchok et al.: Nivolumab plus ipilimumab in advanced melanoma. N Engl J Med 369:(2013)122–133).

[0020] CEACAM6 also helps regulate CD8+T cell responses. In multiple myeloma expressing multiple CEACAM family members, treatment with anti-CEACAM6 mAb or siRNA silencing CEACAM6 restored T cell reactivity against malignant plasma cells, indicating the role of CEACAM6 in CD8+T cell response regulation (Witzens-Harig et al., Blood 2013 May 30; 121(22): 4493-503). WO 2016 / 150899 A2 discloses very effective anti-CECAM6 antibodies for cancer immunotherapy, including TPP-3310.

[0021] definition

[0022] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by those of ordinary skill in the art to which the invention belongs. However, the following references can provide general definitions of many of the terms used in the present invention for those of ordinary skill in the art to which the invention belongs, and as long as these definitions are consistent with the meanings generally understood in the art, they can be referenced and used. Such references include, but are not limited to, Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd edition 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); Hale & Marham, The Harper Collins Dictionary of Biology (1991); and Lackie et al., The Dictionary of Cell & Molecular Biology (3rd edition 1999); and Cellular and Molecular Immunology, Eds. Abbas, Lichtman and Pober, 2nd edition, WB Saunders Company. Any other technical resources available to those of ordinary skill in the art can be consulted, which provide definitions of the terms used herein with the meanings generally understood in the art. For the purposes of the present invention, the following terms are further defined. Other terms are defined elsewhere in the specification. As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, a reference to "a gene" is a reference to one or more genes and includes equivalents thereof known to those skilled in the art, and so forth.

[0023] The terms "polypeptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. These terms apply to amino acid polymers in which one or more amino acid residues is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. Unless otherwise indicated, a particular polypeptide sequence also implicitly includes conservatively modified variants thereof.

[0024] Herein, amino acids may be referred to by either their well-known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Similarly, nucleotides may be referred to by their commonly accepted single-letter codes.

[0025] According to the present invention, the term "antibody" should be understood in its broadest sense and includes immunoglobulin molecules, such as complete or modified monoclonal antibodies, polyclonal antibodies or multispecific antibodies (such as bispecific antibodies). Immunoglobulin molecules are preferably composed of four polypeptide chains, two heavy (H) chains and two light (L) chains, which are usually interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated as VH herein) and a heavy chain constant region. The heavy chain constant region may include, for example, three domains CH1, CH2 and CH3. Each light chain consists of a light chain variable region (abbreviated as VL herein) and a light chain constant region. The light chain constant region consists of one domain (CL). The VH and VL regions can be further subdivided into hypervariable regions, called complementary determining regions (CDRs), with more conservative regions interspersed therebetween, called framework regions (FRs). Each VH and VL is typically composed of three CDRs and up to four FRs, arranged from amino-terminus to carboxyl-terminus, for example, in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0026] As used herein, the term "complementarity determining region" (CDR; e.g., CDR1, CDR2, and CDR3) refers to the amino acid residues of an antibody variable domain whose presence is essential for antigen binding. Each variable domain typically has three CDR regions, identified as CDR1, CDR2, and CDR3. Each complementary determining region may include amino acid residues from the "complementarity determining region" defined by Kabat (e.g., residues 23-34 (L-CDR1), 50-56 (L-CDR2), and 89-97 (L-CDR3) for a light chain variable domain and residues 31-35 (H-CDR1), 50-65 (H-CDR2), and 95-102 (H-CDR3) for a heavy chain variable domain); (Kabat et al., Sequences of Proteins of Immulological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)) and / or those residues from a "hypervariable loop" (e.g., residues 26-32 (L-CDR1), 50-52 (L-CDR2), and 91-96 (L-CDR3) for the light chain variable domain and residues 26-32 (H-CDR1), 53-55 (H-CDR2), and 96-101 (H-CDR3) for the heavy chain variable domain (Chothia and Lesk; J Mol Biol 196:901-917 (1987)). In some cases, the complementarity determining region may include amino acids from both a CDR region and a hypervariable loop according to the Kabat definition.

[0027] Depending on the amino acid sequence of the constant domain of their heavy chains, intact antibodies can be divided into different "classes". There are five major classes of intact antibodies: IgA, IgD, IgE, IgG and IgM, some of which can be further divided into "subclasses" (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. A preferred class of immunoglobulins for use in the present invention is IgG.

[0028] The heavy chain constant domains corresponding to different classes of antibodies are called [α], [δ], [ε], [γ] and [μ], respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. As used herein, antibodies are conventionally known antibodies and functional fragments thereof.

[0029] An "anti-antigen" antibody is an antibody that specifically binds to an antigen. For example, an anti-PD-1 antibody specifically binds to PD-1, and an anti-CECAM6 antibody specifically binds to CECAM6.

[0030] The term "specific binding" or "bind specifically" refers to an antibody or binding agent that binds to a predetermined antigen / target molecule. Specific binding of an antibody or binding agent generally describes a binding affinity of at least 10 -7 M (expressed as Kd value; preferably Kd value is less than 10 -7 M), wherein the affinity of the antibody or binding agent for the predetermined antigen / target molecule is at least two times higher than the affinity for a non-specific antigen / target molecule (e.g., bovine serum albumin or casein) that is not the predetermined antigen / target molecule or a closely related antigen / target molecule. Specific binding of the antibody or binding agent does not exclude binding of the antibody or binding agent to multiple antigens / target molecules (e.g., orthologs of different species). The antibody preferably has at least 10 -7 M (expressed as Kd value; in other words, preferably Kd value is less than 10 -7 M), preferably at least 10 -8 M, more preferably 10 -9 M to 10 -11 The Kd value can be determined, for example, by surface plasmon resonance spectroscopy.

[0031] "Functional fragment", "antigen-binding antibody fragment" or "antibody fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen bound to the whole antibody. "Functional fragment", "antigen-binding antibody fragment" or "antibody fragment" of the present invention includes, but is not limited to, Fab, Fab', Fab'-SH, F(ab')2 and Fv fragments; diabodies; single domain antibodies (DAb), linear antibodies; single-chain antibody molecules (scFv); and multispecific (e.g., bispecific and trispecific) antibodies formed by antibody fragments (CAK Borrebaeck, editor (1995) Antibody Engineering (Breakthroughs in Molecular Biology), Oxford University Press; R. Kontermann & S. Duebel, editors (2001) Antibody Engineering (Springer Laboratory Manual), Springer Verlag).

[0032] The term "immunotherapy" refers to treating a subject having a disease, or at risk of having an infection or having a recurrent disease, by methods that include inducing, enhancing, suppressing or otherwise altering an immune response.

[0033] "Treatment" or "therapy" of a subject refers to any type of intervention or procedure performed on a subject, or the administration of an active agent to a subject, with the purpose of reversing, alleviating, ameliorating, inhibiting, slowing or preventing the onset, progression, development, severity or recurrence of symptoms, complications or conditions associated with a disease, or biochemical markers.

[0034] As used herein, "CEACAM6" refers to "carcinoembryonic antigen-related cell adhesion molecule 6", also known as "CD66c" (cluster of differentiation 66c) or non-specific cross-reactive antigen or NCA or NCA-50 / 90. CEACAM6 is a glycosylphosphatidylinositol (GPI)-linked cell surface protein that is involved in cell-cell adhesion. The term "CEACAM6" as used herein includes human CEACAM6 (hCEACAM6), variants, isoforms and species homologs of hCEACAM6, and analogs that have at least one common epitope with hCEACAM6. The reference sequence of human CEACAM6 can be obtained from the UniProtKB / Swiss-Prot database under accession number P40199.3

[0035] "Programmed death 1 (PD-1)" refers to an immunosuppressive receptor belonging to the CD28 family. PD-1 is primarily expressed on previously activated T cells in vivo and binds to two ligands, PD-L1 and PD-L2. The term "PD-1" as used herein includes human PD-1 (hPD-1), variants, isoforms and species homologs of hPD-1, and analogs that have at least one common epitope with hPD-1. The complete hPD-1 sequence can be found under GenBank accession number U64863.

[0036] "Programmed death ligand-1 (PD-L1)" is one of the two cell surface glycoprotein ligands of PD-1 (the other is PD-L2), which downregulates T cell activation and cytokine secretion when bound to PD-1. The term "PD-L1" as used herein includes human PD-L1 (hPDL1), variants, isoforms and species homologs of hPD-L1, and analogs having at least one common epitope with hPD-L1. The complete hPD-L1 sequence can be found under GenBank accession number Q9NZQ7.

[0037] As used herein, the terms "patient" or "subject" are used interchangeably and refer to mammals, including but not limited to humans or non-human mammals, such as cows, horses, dogs, sheep or cats. Preferably, the patient is a human. DETAILED DESCRIPTION

[0038] Surprisingly, the effects of the combination of a PD-1 or PD-L1 immune checkpoint inhibitor and the anti-CECAM6 antibody TPP-3310 were observed to be greater than additive in in vitro assays conducted to evaluate the therapeutic potential of the drug combination on tumor regression.

[0039] Therefore, the present invention provides a combination of at least two components (component A and component B), component A is the anti-CEACAM6 antibody TPP-3310, and component B is an anti-PD-1 antibody or an anti-PD-L1 antibody, the anti-PD-1 antibody is preferably nivolumab or pembrolizumab, and the anti-PD-L1 antibody is preferably atezolizumab, avelumab or durvalumab.

[0040] As described and defined herein, a combination comprising at least two components A and B is also referred to as a "combination of the invention".

[0041] Furthermore, the present invention relates to a kit comprising:

[0042] - A combination of:

[0043] - Component A, which is the anti-CEACAM6 antibody TPP-3310;

[0044] - component B, which is an anti-PD-1 antibody or an anti-PD-L1 antibody, the anti-PD-1 antibody is preferably nivolumab or pembrolizumab, the anti-PD-L1 antibody is preferably atezolizumab, avelumab or durvalumab, and optionally

[0045] - one or more pharmaceutical preparations C;

[0046] Optionally, one or both of components A and B are in the form of a ready-to-use pharmaceutical preparation for simultaneous, concurrent, separate or sequential administration.

[0047] The present invention further provides an anti-CEACAM6 antibody (component A) for simultaneous, separate or sequential combination with an anti-PD-1 antibody or an anti-PD-L1 antibody (component B) for treating cancer, wherein the anti-CEACAM6 antibody comprises H-CDR1, H-CDR2, H-CDR3, L-CDR1, L-CDR2 and L-CDR3 of antibody TPP-3310.

[0048] The present invention further provides an anti-CEACAM6 antibody (component A), which is used for treating cancer simultaneously, separately or sequentially in combination with an anti-PD-1 antibody or an anti-PD-L1 antibody (component B), wherein the anti-CEACAM6 antibody comprises the variable heavy chain sequence and variable light chain sequence of antibody TPP-3310.

[0049] The present invention further provides an anti-CEACAM6 antibody (component A), which is used for treating cancer simultaneously, separately or sequentially in combination with an anti-PD-1 antibody or an anti-PD-L1 antibody (component B), wherein the anti-CEACAM6 antibody comprises the heavy chain region and the light chain region of the antibody TPP-3310.

[0050] The present invention further provides an anti-CEACAM6 antibody TPP-3310 (component A), which is used for simultaneous, separate or sequential combination with an anti-PD-1 antibody or an anti-PD-L1 antibody (component B) for treating cancer, wherein the anti-PD-1 antibody is nivolumab or pembrolizumab, and the anti-PD-L1 antibody is atezolizumab, avelumab or durvalumab.

[0051] The present invention further provides an anti-CEACAM6 antibody TPP-3310 (component A), which is used for simultaneous, separate or sequential combination with an anti-PD-1 antibody or an anti-PD-L1 antibody (component B) to treat cancer, wherein the cancer is lung cancer, in particular non-small cell lung cancer, ovarian cancer, mesothelioma, pancreatic cancer, gastric cancer, colorectal cancer, head and neck cancer, bladder cancer, bile duct cancer, breast cancer, cervical cancer or esophageal cancer.

[0052] The present invention further provides an anti-CEACAM6 antibody TPP-3310 (component A), which is used for treating cancer simultaneously, separately or sequentially in combination with an anti-PD-1 antibody or an anti-PD-L1 antibody (component B), wherein at least one of the anti-CEACAM6 antibody, anti-PD-1 antibody or anti-PD-L1 antibody is administered simultaneously, separately or sequentially in combination with one or more pharmaceutical preparations (preparation C).

[0053] The present invention further provides a method for treating cancer, comprising administering an effective amount of anti-CEACAM6 antibody TPP-3310 (component A) and anti-PD-1 antibody or anti-PD-L1 antibody (component B) simultaneously, separately or sequentially to a patient in need thereof.

[0054] The present invention further provides a method for treating cancer, comprising administering an effective amount of anti-CEACAM6 antibody TPP-3310 (component A) and anti-PD-1 antibody or anti-PD-L1 antibody (component B) simultaneously, separately or sequentially to a patient in need thereof, wherein the anti-PD-1 antibody is nivolumab or pembrolizumab, and the anti-PD-L1 antibody is atezolizumab, avelumab or durvalumab.

[0055] The present invention further provides the use of anti-CEACAM6 antibody TPP-3310 (component A) in combination with anti-PD-1 antibody or anti-PD-L1 antibody (component B) simultaneously, separately or sequentially for the preparation of a medicament for treating cancer.

[0056] The present invention further provides the use of anti-CEACAM6 antibody TPP-3310 (component A) and anti-PD-1 antibody or anti-PD-L1 antibody (component B) for the preparation of a medicament for treating cancer simultaneously, separately or sequentially, wherein the anti-PD-1 antibody is nivolumab or pembrolizumab, and the anti-PD-L1 antibody is atezolizumab, avelumab or durvalumab. These components can be administered independently of each other by oral, intravenous, topical, local device, intraperitoneal or nasal routes.

[0057] According to another aspect, the present invention encompasses a combination as described above for use in the treatment or prevention of cancer.

[0058] According to another aspect, the present invention encompasses the use of such a combination as described above for the preparation of a medicament for the treatment or prevention of cancer.

[0059] Combination Component A

[0060] Component A is the anti-CEACAM6 antibody TPP-3310 disclosed in WO 2016 / 150899 A2. Other anti-CECAM6 antibodies disclosed in WO 2016150899A2 are, for example, TPP-3714, TPP-3820, TPP-3821, TPP-3707 and TPP-3705. These antibodies are human or humanized antibodies that bind to human CEACAM6 with high affinity, cross-react with monkey CEACAM6, do not bind to any paralogs (paralog) (particularly CEACAM1, CEACAM3 and CEACAM5), and can relieve CEACAM6-mediated immunosuppression.

[0061] The term "anti-CEACAM6 antibody" relates to an antibody that specifically binds to the cancer target molecule CEACAM6, preferably with sufficient affinity for diagnostic and / or therapeutic applications. In certain embodiments, the anti-CEACAM6 antibody binds to an epitope that is conserved between different species.

[0062] TPP-3310 is an antibody comprising an H-CDR1 comprising the amino acid sequence of SEQ ID NO: 12, an H-CDR2 comprising the amino acid sequence of SEQ ID NO: 13, an H-CDR3 comprising the amino acid sequence of SEQ ID NO: 14, an L-CDR1 comprising the amino acid sequence of SEQ ID NO: 16, an L-CDR2 comprising the amino acid sequence of SEQ ID NO: 17, and an L-CDR3 comprising the amino acid sequence of SEQ ID NO: 18.

[0063] Preferably, TPP-3310 is an antibody comprising the variable heavy chain sequence (VH) of SEQ ID NO:11 and the variable light chain sequence (VL) of SEQ ID NO:15.

[0064] Highly preferably, TPP-3310 is an antibody comprising the heavy chain region (HC) of SEQ ID NO:19 and the light chain region (LC) of SEQ ID NO:20.

[0065] Combination Component B

[0066] Component B is an antibody or an antigen-binding portion thereof that specifically binds to the programmed death-1 (PD-1) receptor and inhibits PD-1 activity ("anti-PD-1 antibody"); or an antibody or an antigen-binding portion thereof that specifically binds to the programmed death ligand-1 (PD-L1) and inhibits PD-L1 activity ("anti-PD-L1 antibody").

[0067] Anti-PD-1 antibodies

[0068] In certain embodiments, the anti-PD-1 antibody or its antigen-binding portion is nivolumab or has the same CDR region as nivolumab. Nivolumab (trade name "OPDIVO"; formerly named 5C4, BMS-936558, MDX-1106 or ONO-4538) is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor antibody that selectively prevents interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking the downregulation of anti-tumor T cell function (U.S. Patent No. 8,008,449). In another embodiment, the anti-PD-1 antibody or its fragment cross-competes with nivolumab.

[0069] TPP-2596 is an anti-PD-1 human IgG4 (S228P) antibody cloned using the variable domain of nivolumab.

[0070] In other embodiments, the anti-PD-1 antibody or antigen-binding portion thereof is pembrolizumab or has the same CDR regions as pembrolizumab. Pembrolizumab (trade name "KEYTRUDA", also known as lambrolizumab and MK-3475) is a humanized monoclonal IgG4 antibody against the human cell surface receptor PD-1. For example, pembrolizumab is described in U.S. Pat. No. 8,900,587.

[0071] In other embodiments, the anti-PD-1 antibody or antigen binding portion thereof is MEDI0608 (formerly AMP-514) or has the same CDR regions as MEDI0608. MEDI0608 is a monoclonal antibody directed against the PD-1 receptor. MEDI0608 is described, for example, in U.S. Pat. No. 8,609,089, B2.

[0072] In other embodiments, the anti-PD-1 antibody, or antigen binding portion thereof, is BGB-A317 or has the same CDR regions as BGB-A317. BGB-A317 is a humanized monoclonal antibody described in US Publication No. 2015 / 0079109.

[0073] In certain embodiments, the anti-PD-1 antibody comprises:

[0074] (i) an H-CDR1 comprising the amino acid sequence of SEQ ID NO: 2, an H-CDR2 comprising the amino acid sequence of SEQ ID NO: 3, an H-CDR3 comprising the amino acid sequence of SEQ ID NO: 4, an L-CDR1 comprising the amino acid sequence of SEQ ID NO: 6, an L-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and an L-CDR3 comprising the amino acid sequence of SEQ ID NO: 8, or

[0075] (ii) H-CDR1 comprising the amino acid sequence of SEQ ID NO:32, H-CDR2 comprising the amino acid sequence of SEQ ID NO:33, H-CDR3 comprising the amino acid sequence of SEQ ID NO:34, L-CDR1 comprising the amino acid sequence of SEQ ID NO:36, L-CDR2 comprising the amino acid sequence of SEQ ID NO:37, and L-CDR3 comprising the amino acid sequence of SEQ ID NO:38.

[0076] In certain embodiments, the anti-PD-1 antibody comprises:

[0077] (i) the variable heavy chain sequence (VH) of SEQ ID NO: 1 and the variable light chain sequence (VL) of SEQ ID NO: 5, or

[0078] (ii) the variable heavy chain sequence (VH) of SEQ ID NO:31 and the variable light chain sequence (VL) of SEQ ID NO:35.

[0079] In certain embodiments, the anti-PD-1 antibody comprises:

[0080] (i) the heavy chain region (HC) of SEQ ID NO:9 and the light chain region (LC) of SEQ ID NO:10, or

[0081] (ii) the heavy chain region (HC) of SEQ ID NO:39 and the light chain region (LC) of SEQ ID NO:40.

[0082] Anti-PD-L1 antibodies

[0083] In certain embodiments, the anti-PD-L1 antibody or antigen-binding portion thereof is atezolizumab or has the same CDR regions as atezolizumab. Atezolizumab (trade name "TECENTRIQ"), also known as MPDL3280A, RG7446) is described in U.S. Pat. No. 8,217,149.

[0084] TPP-3615 is an anti-PD-L1 huIgG2 antibody cloned using the variable domain of atezolizumab.

[0085] In other embodiments, the anti-PD-L1 antibody or antigen binding portion thereof is avelumab or has the same CDR regions as avelumab. Avelumab (trade name "BAVENCIO"), also known as MSB0010718C, is described in US 2014 / 0341917.

[0086] In other embodiments, the anti-PD-L1 antibody or antigen-binding portion thereof is durvalumab or has the same CDR regions as durvalumab.Durvalumab (trade name "IMFINZI"), also known as MEDI4736, is described in U.S. Pat. No. 8,779,108 or U.S. 2014 / 0356353.

[0087] In other embodiments, the anti-PD-L1 antibody or antigen binding portion thereof is BMS-936559 or has the same CDR regions as BMS-936559. BMS-936559 (formerly 12A4 or MDX-1105) is a fully human IgG4 monoclonal antibody targeting the PD-1 ligand PD-L1 and is described in U.S. Pat. No. 7,943,743 or WO 2013 / 173223.

[0088] In certain embodiments, the anti-PD-L1 antibody comprises:

[0089] (iii) an H-CDR1 comprising the amino acid sequence of SEQ ID NO:42, an H-CDR2 comprising the amino acid sequence of SEQ ID NO:43, an H-CDR3 comprising the amino acid sequence of SEQ ID NO:44, an L-CDR1 comprising the amino acid sequence of SEQ ID NO:46, an L-CDR2 comprising the amino acid sequence of SEQ ID NO:47, and an L-CDR3 comprising the amino acid sequence of SEQ ID NO:48, or

[0090] (iv) an H-CDR1 comprising the amino acid sequence of SEQ ID NO:52, an H-CDR2 comprising the amino acid sequence of SEQ ID NO:53, an H-CDR3 comprising the amino acid sequence of SEQ ID NO:54, an L-CDR1 comprising the amino acid sequence of SEQ ID NO:56, an L-CDR2 comprising the amino acid sequence of SEQ ID NO:57, and an L-CDR3 comprising the amino acid sequence of SEQ ID NO:58, or

[0091] (v) an H-CDR1 comprising the amino acid sequence of SEQ ID NO: 62, an H-CDR2 comprising the amino acid sequence of SEQ ID NO: 63, an H-CDR3 comprising the amino acid sequence of SEQ ID NO: 64, an L-CDR1 comprising the amino acid sequence of SEQ ID NO: 66, an L-CDR2 comprising the amino acid sequence of SEQ ID NO: 67, and an L-CDR3 comprising the amino acid sequence of SEQ ID NO: 68, or

[0092] (vi) H-CDR1 comprising the amino acid sequence of SEQ ID NO:22, H-CDR2 comprising the amino acid sequence of SEQ ID NO:23, H-CDR3 comprising the amino acid sequence of SEQ ID NO:24, L-CDR1 comprising the amino acid sequence of SEQ ID NO:26, L-CDR2 comprising the amino acid sequence of SEQ ID NO:27, and L-CDR3 comprising the amino acid sequence of SEQ ID NO:28.

[0093] In certain embodiments, the anti-PD-L1 antibody comprises:

[0094] (iii) the variable heavy chain sequence (VH) of SEQ ID NO:41 and the variable light chain sequence (VL) of SEQ ID NO:45, or

[0095] (iv) the variable heavy chain sequence (VH) of SEQ ID NO:51 and the variable light chain sequence (VL) of SEQ ID NO:55, or

[0096] (v) the variable heavy chain sequence (VH) of SEQ ID NO:61 and the variable light chain sequence (VL) of SEQ ID NO:65, or

[0097] (vi) the variable heavy chain sequence (VH) of SEQ ID NO:21 and the variable light chain sequence (VL) of SEQ ID NO:25.

[0098] In certain embodiments, the anti-PD-L1 antibody comprises:

[0099] (iii) the heavy chain region (HC) of SEQ ID NO:49 and the light chain region (LC) of SEQ ID NO:50, or

[0100] (iv) the heavy chain region (HC) of SEQ ID NO:59 and the light chain region (LC) of SEQ ID NO:60, or

[0101] (v) the heavy chain region (HC) of SEQ ID NO:69 and the light chain region (LC) of SEQ ID NO:70, or

[0102] (vi) the heavy chain region (HC) of SEQ ID NO:29 and the light chain region (LC) of SEQ ID NO:30.

[0103] Antibody production

[0104] Antibodies or antigen-binding antibody fragments binding to target molecules can be prepared by those of ordinary skill in the art using known methods (e.g., chemical synthesis or recombinant expression). Binding agents for cancer target molecules are commercially available or can be prepared by those of ordinary skill in the art using known methods (e.g., chemical synthesis or recombinant expression). Other methods for preparing antibodies or antigen-binding antibody fragments are described in WO 2007 / 070538 (see page 22, "antibodies"). Those skilled in the art know how to compile, for example, phage display libraries (e.g., Morphosys HuCAL Gold), and for discovering the process of antibodies or antigen-binding antibody fragments (see WO 2007 / 070538, page 24 and subsequent pages, and AK Example 1 on page 70, AK Example 2 on page 72). For example, other methods for preparing antibodies using the DNA library of B cells are described on page 26 (WO 2007 / 070538). Methods for humanizing antibodies are described on pages 30-32 of WO2007070538 and described in detail in Queen et al., Pros. Natl. Acad. Sci. USA 8610029-10033, 1989 or WO 90 / 0786.In addition, methods for recombinant expression of proteins are generally known to those skilled in the art, and methods for recombinant expression of antibodies are particularly known (see, for example, Berger and Kimmel (Guide to Molecular Cloning Techniques, Methods in Enzymology, Vol. 152, Academic Press, Inc.); Sambrook et al., (Molecular Cloning A Laboratory Manual, (2nd Edition, Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY; 1989) Vols. 1-3); Current Protocols in Molecular Biology, (FM Ausabel et al. [Eds.], Current Protocols, Green Publishing Associates, Inc. / John Wiley & Sons, Inc.); Harlow et al., (Monoclonal Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory Press (19881, Paul [Ed.]); Fundamental Immunology, (Lippincott Williams & Wilkins (1998)); and Harlow et al., (Using Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory Press (1998)). The corresponding vectors, promoters and signal peptides are known to those skilled in the art, which are necessary for protein / antibody expression. Common methods are also described in WO 2007 / 070538, pages 41-45. For example, methods for preparing IgG1 antibodies are described in Example 6 on pages 74 and thereafter of WO 2007 / 070538. Technicians are known, and methods that allow determination of the internalization of antibodies after the antibodies bind to their antigens are described, for example, on page 80 of WO 2007 / 070538. Those skilled in the art can use the methods described in WO 2007 / 070538, which have been used to prepare carbonic anhydrase IX (Mn) antibodies, similar to antibodies used to prepare antibodies with different target molecule specificities.

[0105] Bacterial expression

[0106] Those skilled in the art are aware of the ways in which antibodies, antigen-binding fragments thereof, or variants thereof can be produced via bacterial expression.

[0107] By inserting a DNA sequence and suitable translation initiation and translation termination signals and a functional promoter, a suitable expression vector for bacterial expression of the desired protein is constructed, the DNA sequence encoding the desired protein in a functional reading frame. The vector comprises one or more phenotypic selectable markers and a replication origin, so that the vector can be retained, and if necessary, amplified in the host. Suitable prokaryotic hosts for conversion include but are not limited to Escherichia coli (E.coli), Bacillus subtilis (Bacillus subtilis), Salmonella typhimurium (Salmonella typhimurium) and various species from Pseudomonas (Pseudomonas), Streptomyces (Streptomyces) and Staphylococcus (Staphylococcus). Bacterial vectors can be based on, for example, phages, plasmids or phagemids. These vectors can contain selectable markers and bacterial replication origins, which are derived from commercially available plasmids. Many commercially available plasmids usually contain elements of the well-known cloning vector pBR322 (ATCC 37017). In bacterial systems, many favorable expression vectors can be selected based on the intended use of the protein to be expressed.

[0108] After transformation of a suitable host strain and growth of the host strain to an appropriate cell density, the selected promoter is de-reprimed / induced by a suitable method (e.g., temperature shift or chemical induction), and the cells are cultured for an additional period of time. The cells are typically collected by centrifugation and, if necessary, physically or chemically digested, and the resulting crude extract is retained for further purification.

[0109] Mammalian cell expression

[0110] Those skilled in the art are aware of the ways in which antibodies, antigen-binding fragments thereof, or variants thereof can be produced via mammalian cell expression.

[0111] Preferred regulatory sequences for expression in mammalian cell hosts include viral elements that result in high expression in mammalian cells, such as promoters and / or expression amplifiers derived from cytomegalovirus (CMV) (e.g., CMV promoter / enhancer), simian virus 40 (SV40) (e.g., SV40 promoter / enhancer), adenovirus (e.g., adenovirus major late promoter (AdMLP)), and polyomavirus. Expression of the antibody can be constitutive or regulated (e.g., induced by addition or removal of small molecule inducers that bind to the Tet system (e.g., tetracycline)).

[0112] To further describe viral regulatory elements and sequences thereof, reference is made to, for example, Stinski's U.S. 5,168,062, Bell et al.'s U.S. 4,510,245 and Schaffner et al.'s U.S. 4,968,615. Recombinant expression vectors may also include an origin of replication and a selectable marker (see, for example, U.S. 4,399,216, 4,634,665 and U.S. 5,179,017). When the vector is introduced into a cell, suitable selectable markers include genes that confer resistance to substances (e.g., G418, puromycin, hygromycin, blasticidin, zeocin / bleomycin or methotrexate) or selectable markers that cause auxotrophic defects in host cells, such as glutamine synthetase (Bebbington et al., Biotechnology (NY). 1992 Feb; 10 (2): 169-75).

[0113] For example, the dihydrofolate reductase (DHFR) gene confers resistance to methotrexate, the neo gene confers resistance to G418, the bsd gene of Aspergillus terreus confers resistance to blasticidin, puromycin N-acetyltransferase confers resistance to puromycin, the Sh ble gene product confers resistance to bleomycin, and the Escherichia coli hygromycin resistance gene (hyg or hph) confers resistance to hygromycin. Selectable markers (e.g., DHFR or glutamine synthetase) also facilitate amplification techniques used in conjunction with MTX and MSX.

[0114] Expression vectors can be transfected into host cells by standard techniques including electroporation, nucleofection, calcium phosphate precipitation, lipofection, polycation-based transfection such as polyethyleneimine (PEI)-based transfection, and DEAE-dextran transfection.

[0115] Suitable mammalian host cells for expressing antibodies, antigen-binding fragments thereof, or variants thereof include Chinese hamster ovary (CHO) cells, such as CHO-K1, CHO-S, CHO-K1SV [including DHFR-CHO cells, described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220 and Urlaub et al., Cell. 1983 Jun; 33(2):405-12, used with a DHFR-selectable marker, such as described in RJ Kaufman and Sharp (1982) Mol. Biol. 159:601-621, and other knockout cells, such as Fan et al., Biotechnol Bioeng. 2012 Apr; 109(4): 1007-15), NS0 myeloma cells, COS cells, HEK293 cells, HKB11 cells, BHK21 cells, CAP cells, EB66 cells and SP2 cells.

[0116] The expression of the antibody, antigen-binding fragment thereof or variant thereof can also be affected in a transient or semi-stable manner in an expression system, such as HEK293, HEK293T, HEK293-EBNA, HEK293E, HEK293-6E, HEK293Freestyle, HKB11, Expi293F, 293EBNALT75, CHO Freestyle, CHO-S, CHO-K1, CHO-K1SV, CHOEBNALT85, CHOS-XE, CHO-3E7 or CAP-T cells (e.g. Durocher et al., Nucleic Acids Res. 2002 Jan 15; 30(2):E9).

[0117] In some embodiments, the expression vector is constructed in such a way that the protein to be expressed is secreted into the cell culture medium in which the host cells are grown. The antibody, antigen-binding fragment thereof, or variant thereof can be obtained from the cell culture medium by protein purification methods known to those skilled in the art.

[0118] purification

[0119] Antibodies, antigen-binding fragments thereof, or variants thereof can be obtained and purified from recombinant cell cultures by well-known methods, examples of which include ammonium sulfate or ethanol precipitation, acid extraction, protein A chromatography, protein G chromatography, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography (HIC), affinity chromatography, hydroxyapatite chromatography, and lectin chromatography. High performance liquid chromatography ("HPLC") can also be used for purification. See, e.g., Colligan, Current Protocols in Immunology, or Current Protocols in Protein Science, John Wiley & Sons, NY, NY, (1997-2001), e.g., Chapters 1, 4, 6, 8, 9, 10.

[0120] The antibodies or antigen-binding fragments thereof or variants thereof of the present invention include naturally purified products, products from chemical synthesis methods, and products produced in prokaryotic or eukaryotic host cells by means of recombinant technology. Eukaryotic hosts include, for example, yeast cells, higher plant cells, insect cells, and mammalian cells. Depending on the host cell selected for recombinant expression, the expressed protein can be in glycosylated or non-glycosylated form.

[0121] In a preferred embodiment, the antibody is purified (1) to a degree greater than 95% by weight, for example, as measured by the Lowry method, by UV-visible spectroscopy or by SDS capillary gel electrophoresis (e.g., with a Caliper LabChip GXII, GX 90 or Biorad Bioanalyzer instrument), in a more preferred embodiment, greater than 99% by weight, (2) to a degree suitable for determination of at least 15 residues of N-terminal or internal amino acid sequence, or (3) to homogeneity as determined by SDS-PAGE under reducing or non-reducing conditions with the aid of Coomassie blue or, preferably, silver staining.

[0122] Ordinarily, isolated antibodies are obtained with the aid of at least one protein purification step.

[0123] Table 1: Protein sequences of preferred antibodies

[0124]

[0125] Table 2: Sequences of preferred antibodies

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134] Methods of treating cancer

[0135] In the context of the present invention, the term "cancer" includes, but is not limited to, breast cancer, lung cancer, brain cancer, reproductive organ cancer, digestive tract cancer, urinary tract cancer, liver cancer, eye cancer, skin cancer, head and neck cancer, thyroid cancer, parathyroid cancer and distant metastasis thereof. Those diseases also include multiple myeloma, lymphoma, sarcoma and leukemia.

[0136] Examples of breast cancer include, but are not limited to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ.

[0137] Examples of respiratory tract cancers include, but are not limited to, lung cancer, particularly small cell and non-small cell lung cancer, as well as bronchial adenoma and pleuropulmonary blastoma.

[0138] Examples of brain cancers include, but are not limited to, brainstem and pituitary gliomas, cerebellar and cerebral astrocytomas, medulloblastomas, ependymomas, and neuroectodermal and pineal tumors.

[0139] Tumors of male reproductive organs include, but are not limited to, prostate cancer and testicular cancer. Tumors of female reproductive organs include, but are not limited to, endometrial cancer, cervical cancer, ovarian cancer, vaginal cancer, vulvar cancer, and uterine sarcoma.

[0140] Digestive tract tumors include, but are not limited to, anal cancer, colon cancer, colorectal cancer, esophageal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small intestine cancer, and salivary gland cancer.

[0141] Urinary tract tumors include, but are not limited to, bladder cancer, penile cancer, kidney cancer, renal pelvis cancer, ureter cancer, urethral cancer, and human papillary renal carcinoma.

[0142] Eye cancers include, but are not limited to, intraocular melanoma and retinoblastoma.

[0143] Examples of liver cancer include, but are not limited to, hepatocellular carcinoma (liver cell carcinoma with or without fibrolamellar variant), cholangiocarcinoma (intrahepatic bile duct carcinoma), and mixed hepatocellular cholangiocarcinoma.

[0144] Skin cancers include, but are not limited to, squamous cell carcinoma, Kaposi's sarcoma, melanoma, particularly malignant melanoma, Merkel cell skin cancer, and non-melanoma skin cancer.

[0145] Head and neck cancers include, but are not limited to, cancers of the larynx, hypopharynx, nasopharynx, oropharynx, lip and oral cavity, and squamous cell carcinoma.

[0146] Lymphomas include, but are not limited to, AIDS-related lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Burkitt's lymphoma, Hodgkin's disease, and central nervous system lymphoma.

[0147] Sarcomas include, but are not limited to, soft tissue sarcomas, osteosarcomas, malignant fibrous histiocytomas, lymphosarcoma, and rhabdomyosarcomas.

[0148] Leukemias include, but are not limited to, acute myeloid leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, and hairy cell leukemia.

[0149] The present invention relates to a method for treating or preventing cancer using the combination of the present invention, in particular (but not limited to) colorectal cancer, lung cancer, pancreatic cancer, breast cancer, prostate cancer, bladder cancer, gastric cancer, head and neck cancer, liver cancer, brain cancer, melanoma, endometrial cancer, lymphoma, leukemia, etc. The combination can be used to inhibit, block, reduce, decrease (etc.) cell proliferation and / or cell division, and / or induce apoptosis in the treatment or prevention of cancer, in particular (but not limited to) colorectal cancer, lung cancer, breast cancer, prostate cancer, bladder cancer, gastric cancer, head and neck cancer, liver cancer, brain cancer, melanoma, endometrial cancer, lymphoma, leukemia, etc. The method comprises administering to a mammal (including a human) in need thereof a certain amount of the combination of the present invention or a pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate, solvate or ester thereof; etc., which is effective for treating or preventing cancer, particularly (but not limited to) colorectal cancer, lung cancer, pancreatic cancer, breast cancer, prostate cancer, bladder cancer, gastric cancer, head and neck cancer, liver cancer, brain cancer, melanoma, endometrial cancer, lymphoma, leukemia, etc.

[0150] The terms "treating" or "treatment" as described herein are generally used, for example, to manage or care for a subject for the purpose of combating, alleviating, reducing, alleviating, improving the condition of a disease or disorder (eg, cancer), and the like.

[0151] In a preferred embodiment, the cancer is lung cancer, in particular non-small cell lung cancer (NSCLC), ovarian cancer, mesothelioma, pancreatic cancer or gastric cancer, colorectal cancer, head and neck cancer, bladder cancer, bile duct cancer, breast cancer, cervical cancer, esophageal cancer.

[0152] Dosage and administration

[0153] Based on known standard laboratory techniques for evaluating compounds useful for treating or preventing cancer, particularly (but not limited to) colorectal cancer, lung cancer, pancreatic cancer, breast cancer, prostate cancer, bladder cancer, gastric cancer, head and neck cancer, liver cancer, brain cancer, melanoma, endometrial cancer, lymphoma, leukemia, etc., by standard toxicity tests and standard pharmacological tests to determine the treatment of the above-identified conditions in mammals, and by comparing these results with the results of known drugs for treating these conditions, the effective dosage of the combination of the present invention can be readily determined for the treatment of the indicated condition. Depending on such considerations as the particular combination and dosage unit used, the mode of administration, the duration of treatment, the age, weight and sex of the patient being treated, and the nature and extent of the condition being treated, the amount of active ingredient administered in treating the condition can vary widely.

[0154] The total amount of active ingredient to be administered will generally be from about 0.001 mg / kg body weight to about 200 mg / kg body weight per day, and preferably from about 0.01 mg / kg body weight to about 30 mg / kg body weight per day. Clinically useful dosing schedules range from one to three times a day to once every four weeks. In addition, a "drug holiday", where patients are not given medication for a period of time, may be beneficial to the overall balance between pharmacological action and tolerance. A unit dose may contain about 0.5 mg to about 2,500 mg of active ingredient, and may be administered once or more or less than once a day. The average dose administered by injection (including intravenous, intramuscular, subcutaneous and parenteral injection) and the average dose administered using an infusion technique are preferably 0.01 to 200 mg / kg total body weight.

[0155] Of course, the specific initial and continued dosing regimen for each patient will vary depending on the nature and severity of the condition as determined by the attending diagnostician, the activity of the specific combination employed, the age, weight and general condition of the patient, the time of administration, the route of administration, the rate of drug excretion, the drug combination, etc. The desired treatment pattern and number of doses of the combination of the present invention, or a pharmaceutically acceptable salt or ester or composition thereof, can be determined by those skilled in the art using routine therapeutic testing.

[0156] A therapy using a combination of component A as described above, component B as described above and component C: one or more other pharmaceutical agents.

[0157] The combination of component A and component B of the present invention can be administered as a sole pharmaceutical preparation or in combination with one or more other pharmaceutical preparations, wherein the resulting combination of components A, B and C does not cause unacceptable adverse effects. For example, the combination of components A and B of the present invention can be combined with component C (i.e., one or more other pharmaceutical preparations), such as known anti-angiogenic, anti-hyperproliferative, anti-inflammatory, analgesic, immunomodulatory, diuretic, anti-arrhythmic, anti-hypercholesterolemia, anti-dyslipidemia, anti-diabetic or antiviral preparations, and mixtures and combinations thereof.

[0158] Component C can be one or more pharmaceutical preparations, such as 131I-chTNT, abarelix, abiraterone, aclarubicin, adalimumab, ado-trastuzumab emtansine, afatinib, aflibercept, aldesleukin, alectinib, alemtuzumab, alendronic acid, acid, alitretinoin, altretamine, amifostine, aminoglutethimide, hexylaminolevulinate, amrubicin, amsacrine, anastrozole, ancestim, anethole dithiolethione, anetumab ravtansine, angiotensin II, antithrombin III, aprepitant, arcitumomab, arglabin, arsenic trioxide trioxide), asparaginase, atezolizumab, axitinib, azacitidine, basiliximab, belotecan, bendamustine, besilesomab, belinostat, bevacizumab, bexarotene, bicalutamide, bisantrene, bleomycin, blinatumomab, bortezomib, buserelin, bosutinib, brentuximabvedotin, busulfan, cabazitaxel, cabozantinib, calcitonine, calcium folinate, calcium folinate levofolinate, capecitabine, capromab, carbamazepine, carboplatin, carboquone, carfilzomib, carmofur, carmustine, catumaxomab, celecoxib, celmoleukin, ceritinib, cetuximab, chlorambucil, chlormadinone, chlormethine, cidofovir, cinacalcet, cisplatin, cladribine, clodronic acid acid, clofarabine, cobimetinib, copanlisib, crisantaspase, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daratumumab, darbepoetin alfaalfa), dabrafenib, dasatinib, daunorubicin, decitabine, degarelix, denileukindiftitox, denosumab, depreotide, deslorelin, dianhydrogalactitol, dexrazoxane, dibrospidiumchlor ide), dianhydrogalactitol, diclofenac, dinutuximab, docetaxel, dolasetron, doxifluridine, doxorubicin, doxorubicin + estrone, dronabinol, eculizumab, edrecolomab, elliptinium acetate, elotuzumab, eltrombopag, endostatin, enocitabine, enzalutamide, epirubicin, epitiostanol, epoetin alfa, epoetin beta, epoetin zeta zeta), eptaplatin, eribulin, erlotinib, esomeprazole, estradiol, estramustine, ethinylestradiol, etoposide, everolimus, exemestane, fadrozole, fentanyl, filgrastim, fluoxymesterone, floxuridine, fludarabine, fluorouracil, flutamide, folinic acidacid, formestane, fosaprepitant, fotemustine, fulvestrant, gadobutrol, gadoteridol, gadotericacid meglumine, gadooversetamide, gadoxetic acid, gallium nitrate, ganirelix, gefitinib, gemcitabine, gemtuzumab, Glucarpidase, glutoxim, GM-CSF, goserelin, granisetron, granulocyte colony stimulating factor factor), histamine dihydrochloride, histrelin, hydroxycarbamide, I-125 seed, lansoprazole, ibandronic acid, ibritumomab tiuxetan, ibrutinib, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, indisetron, incadronic acid, ingenol mebutate, interferon alfa, interferon beta, interferon gammagamma), iobitridol, iobenguane (123I), iomeprol, ipilimumab, irinotecan, itraconazole, ixabepilone, ixazomib, lanreotide, lansoprazole, lapatinib, lasocholine, lenalidomide, lenvatinib, lenograstim, lentinan, letrozole, leuprorelin, levamisole, levonorgestrel, levothyroxine sodium), lisuride, lobaplatin, lomustine, lonidamine, masoprocol, medroxyprogesterone, megestrol, melarsoprol, melphalan, mepitiostane, mercaptopurine, mesna, methadone, methotrexate, methoxsalen, methylaminolevulinate ), methylprednisolone, methyltestosterone, metirosine, mifamurtide, miltefosine, miriplatin, mitobronitol, mitoguazone, mitolactol, mitomycin, mitotane, mitoxantrone, mogamulizumab, molgramostim, mopidamol, morphine hydrochloride, morphine sulfatesulfate), nabilone, nabiximols, nafarelin, naloxone + pentazocine, naltrexone, nartograstim, necitumumab, nedaplatin, nelarabine, neridronic acid, netupitant / palonosetron, nivolumab, pentreotide tetreotide, nilotinib, nilutamide, nimorazole, nimotuzumab, nimustine, nintedanib, nitracrine, nivolumab, obinutuzumab, octreotide, ofatumumab, olaparib, olaratumab, omacetaxine mepesuccinate, omeprazole, ondansetron, oprelvekin, orgotein, orilotimod, osimertinib, oxaliplatin, oxycodone, oxymetholone, ozogamicine, p53 gene therapy, paclitaxel, palbociclib, palifermin, palladium-103seed, palonosetron, pamidronate acid), panitumumab, panobinostat, pantoprazole, pazopanib, pegaspargase, PEG-epoetin beta (methoxy PEG-epoetin beta)beta), pembrolizumab, pegfilgrastim, peginterferon alpha-2b alfa-2b), pembrolizumab, pemetrexed, pentazocine, pentostatin, peplomycin, perflubutane, perfosfamide, pertuzumab, picibanil, pilocarpine, pirarubicin, pixantrone, plerixafor, plicamycin, poliglusam, polyestradiolphosphate, polyvinylpyrrolidone + sodium hyaluronate, polysaccharide-K, pomalidomide, ponatinib, porfimer sodium sodium), pralatrexate, prednimustine, prednisone, procarbazine, procodazole, propranolol, quinagolide, rabeprazole, racotumomab, radium-223 chloride, radotinib, raloxifene, raltitrexed, ramosetron, ramucirumab, ranimustine, rasburicase, razoxane, refametinib, regorafenib, risedronic acid,acid), rhenium-186 etidronate, rituximab, rolapitant, romidepsin, romiplostim, romurtide, roniciclib, rucaparib, samarium (153Sm) lexidronam, sargramostim, satumomab, secretin, siltuximab, sipuleucel-T, sizofiran, sobuzoxane, sodium glycidazole glycididazole), sonidegib, sorafenib, stanozolol, streptozocin, sunitinib, talaporfin, talimogene laherparepvec, tamibarotene, tamoxifen, tapentadol, tasonermin, teceleukin, technetium (99mTc) nofetumomab merpentan), 99mTc-HYNIC-[Tyr3]-octreotide, tegafur, tegafur + gimeracil + oteracil, temoporfin, temozolomide, temsirolimus, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thyrotropin αalfa), tioguanine, tocilizumab, topotecan, toremifene, tositumomab, trabectedin, trametinib, tramadol, trastuzumab, trastuzumab-emtansine emtansine, treosulfan, tretinoin, trifluridine + tipiracil, trilostane, triptorelin, trametinib, trofosfamide, thrombopoietin, tryptophan, ubenimex, valatinib, valrubicin, vandetanib nib), vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vismodegib, vorinostat, vorozole, yttrium-90 glass microspheres, zinostatin, zinostatin stimalamer, zoledronic acid, zorubicin.

[0159] Typically, using component C in conjunction with the combination of components A and B of the present invention will help:

[0160] (1) Compared with either agent given alone, it produces better efficacy in reducing tumor growth or even eliminating tumors,

[0161] (2) provide for administration of a smaller amount of the chemotherapeutic agent being administered,

[0162] (3) provide patients with chemotherapy treatment that is well tolerated and has fewer adverse pharmacologic complications than those observed with single-agent chemotherapy and certain other combination therapies,

[0163] (4) provide treatment for a wider spectrum of different cancer types in mammals, especially humans,

[0164] (5) provide higher response rates among treated patients,

[0165] (6) provide longer survival times in treated patients compared to standard chemotherapy treatment,

[0166] (7) provide a longer time for tumor progression, and / or

[0167] (8) produce efficacy and tolerability results that are at least as good as those of the agents alone, compared with known situations where the combination produces antagonistic effects with other cancer agents.

[0168] The following examples illustrate the feasibility of the present invention, but the present invention is not limited to these examples.

[0169] Example

[0170] Effect of combination therapy of TPP-3310 antibody against human CEACAM6 with antibodies against PD-L1 or PD-1 on activation of PD-1-positive viral peptide-specific T cells

[0171] Since CEACAM6 is not expressed in rodents (no rodent ortholog), in vivo efficacy studies are not possible and preclinical in vivo combination studies cannot be performed to evaluate the therapeutic potential of drug combinations.

[0172] Alternatively, an in vitro cellular assay system was established to test the in vitro efficacy and therapeutic potential of combinations of antibodies against CEACAM6 and PD-1 or PD-L1.

[0173] In this cell test system, PD-1 positive FluM1 viral peptide specific T cells were used as effector T cells. They were co-cultured with PD-L1 and CEACAM6 positive and FLuM1 peptide loaded cancer cells HCC2935 for 24-48 hours in the presence of checkpoint inhibitory antibodies against CEACAM6, PD-1 or PD-L1 (as single agents or in combination). The induction of pro-inflammatory cytokines (IFNg) was measured as a readout of efficacy.

[0174] Antibody

[0175] The antibodies used are TPP-3310 (anti-CEACAM6), a huIgG2 antibody against the immune checkpoint molecule CEACAM6 overexpressed on cancer cells and myeloid cells; TPP-3615, an anti-PD-L1 huIgG2 antibody cloned using the variable domain of atezolizumab; and TPP-2596, an anti-PD-1 human IgG4 (S228P) antibody cloned using the variable domain of nivolumab. TPP-1238 (huIgG2) and TPP1240 (huIgG4) have been used as isotype control antibodies.

[0176] Cell lines and culture

[0177] HCC2935 cancer cells (ATCC-CRL-2869, lung adenocarcinoma) were cultured in RPMI-1640, 10% FCS, 5% CO2. CEACAM6 and PD-L1 expression were confirmed by FACS analysis. For co-culture experiments with viral peptide-specific T cells, cancer cells were pulsed with 0.2 μg / ml or as indicated viral FluM1 peptide.

[0178] Generation and cell culture of FluM1 viral peptide-specific T cells

[0179] PD-1 expressing viral (influenza) peptide-specific T cells from HLA-A*0201 + Initial PBMC generation from healthy donors obtained by Ficoll density centrifugation of buffy coats (Deutsches Rotes Kreuz, Mannheim). CD8 + T cells. CD8 negative cells were irradiated (35Gy) and pulsed with 1 μg / ml of influenza HLA-A*0201 epitope GILGFVFTL (ProImmune) for 1.5 hours at 37°C in X-Vivo-20 medium (chemically defined serum-free hematopoietic cell culture medium, Lonza, #BE04-448Q), followed by washing. Cells were restimulated with irradiated T2 cells and pulsed with 1 μg / ml of its related GILGFVFTL peptide on the 7th day. On the 14th day, aliquots were frozen. The samples were thawed and washed immediately, and then used for functional tests. Before the co-culture experiment on the 14th day, the suitability of viral peptide-specific T cells was confirmed with tetramer (F391-4A-E, ProImmune) staining and FACS analysis.

[0180] In vitro assay: Analysis of the efficacy of combination antibodies in co-culture of T cells and cancer cells

[0181] For co-culture, cancer cells were non-enzymatically detached with PBS-EDTA for 5-15 minutes, centrifuged at 1,400 rpm for 5 minutes, washed and counted. Cancer cells were diluted to 1×10 in X-Vivo-20 (Lonza, #BE04-448Q). 5 cells / ml, pretreated with TPP-3310, aPD-L1 and / or isotype control antibodies for 10 min on ice. After incubation, 10,000 target cancer cells were seeded in triplicate into 96-well ELISA U plates.

[0182] Viral peptide-specific T cells were collected, washed with X-Vivo-20, and diluted to 2x10 in X-Vivo-20. 5 cells / ml and pretreated with anti-PD-1 or isotype control antibodies for 10 minutes on ice. All antibodies were applied at a final concentration of 30 μg / ml. For combination treatment, TPP-3310 was applied at approximately its half-maximal effective concentration (EC50) of 1 μg / ml to ensure the effect of other antibodies on T cell activation. Pretreated T cells were seeded onto target cancer cells at 20,000 cells / well.

[0183] The co-culture of cancer cells and effector T cells with antibodies was incubated at 37°C, 5% CO2 for approximately 20 hours.

[0184] The supernatant was then collected and the co-culture plate was centrifuged at 1400 rpm for 3 minutes. The IFN-γ level in the supernatant was measured by ELISA (human IFN-γ-ELISA device, BD, #555142) according to the manufacturer's instructions. The optical density of the ELISA plate was measured using a Tecan Infinite M200 microplate reader.

[0185] Data were statistically analyzed using paired or unpaired two-tailed Student's t-test using Microsoft Excel 2010 and GraphPad Prism 6. Results with p < 0.05 were considered significant. Cytokine concentrations were calculated using standard curves. Factors or ratios were calculated by dividing the value of TPP-3310 or a given combination by the value of the respective isotype control.

[0186] result

[0187] Before the experiment, HCC2935 cancer cells loaded with FLuM1 peptide were co-incubated with FluM1 viral peptide-specific T cells. Only in the presence of homologous viral peptides, IFN-γ secretion of T cells increased. This increase is dose-dependent. In the presence of anti-CEACAM6 antibody TPP-3310, anti-PD-L1 antibody TPP-3615 or in the presence of anti-PD-1 antibody TPP-2596, IFN-γ secretion in co-culture (p<0.05 to 0.0001) was further enhanced. All were administered as a single preparation. These data confirm that the newly established cell test system consisting of PD-1-positive FluM1 viral peptide-specific T cells and HCC2935 cancer cells loaded with peptides is suitable for testing the efficacy of anti-CEACAM6, anti-PD-1 and anti-PD-L1 antibodies in benchmarking and combination experiments.

[0188] Table 3: Peptide specificity of viral peptide-specific T cell activation measured by IFNg secretion in co-culture experiments with HCC2935 cancer cells loaded with viral peptides with or without immune checkpoint blocking antibodies against CEACAM6, PD-1, or PD-L1.

[0189]

[0190] Instructions: Viral peptide-specific T cells (TC) were stimulated with HCC2935 lung cancer cells (HCC) pulsed with serially diluted viral peptides. 30 μg / ml of antibody was added. The concentration of secreted IFN-γ was determined by ELISA. Data are absolute amounts of IFN-γ in pg / ml. TPP-3310, aCEACAM6; TPP-3615, aPD-L1; TPP-2596, aPD1; TPP-1238, isotype control for TPP-3310 and TPP3615, aPD-1; TPP-1240, isotype control for TPP-2596.

[0191] In 7 independent co-culture experiments (n=7), the effects of the combination of the anti-CEACAM6 antibody TPP-3310 and the antibody against PD-L1 were determined overall. In the presence of the antibodies, we consistently found that IFNg secretion (absolute mean) increased when administered as a single formulation or in combination. In the presence of the PD-L1 antibody, total IFNg increased by 39.6pg / ml, in the presence of the anti-CEACAM6 antibody TPP-3310, it increased by 196.6pg / ml, and when administered in combination, it increased by 279.9pg / ml. This result shows that when the PD-L1 antibody is combined with the CEACAM6 antibody, IFNg secretion is further enhanced, and the effect on IFNg secretion is greater than the additive effect.

[0192] Table 4: Total IFNg secretion in co-culture experiments of FluM1 viral peptide-specific T cells and HCC2935 cells loaded with FluM1 peptide in the presence of anti-CEACAM6 and anti-PD-L1 antibodies (as single agents or in combination) (n=7)

[0193]

[0194] Description table: HCC2935 lung cancer cells (HCC) were pulsed with 0.2 μg / ml FluM1 peptide to stimulate viral peptide-specific T cells (TC) in co-culture. 30 μg / ml of antibody was applied. For combination treatment (n=7), 1 μg / ml of TPP-3310 was added. The concentration of secreted IFN-γ was determined by ELISA, and the data are isotype-corrected values ​​and expressed in pg / ml. TPP-3310, aCEACAM6; TPP-3615, aPD-L1; T-test of means, p-values ​​(<0.05): aPD-L1 vs. CEACAM6, p=0.0439; aPD-L1 vs. combination, p=0.001; aCEACAM6 vs. combination, p=0.16

[0195] In another study, we determined the effect of the combination of the anti-CEACAM6 antibody TPP-3310 and an antibody against PD-1 in a total of 7 independent co-culture experiments (n=7). In the presence of the antibodies, we consistently found an increase in IFNg secretion (absolute mean) when administered as a single formulation or in combination.

[0196] In the presence of PD-1 antibody, the average total IFNg increased by 76.1 pg / ml, in the presence of anti-CEACAM6 antibody TPP-3310, it increased by 166.8 pg / ml, and when administered in combination, it increased by 317.9 pg / ml. This result shows that when PD-1 antibody is combined with CEACMA6 antibody, IFNg secretion is further enhanced, and the effect on IFNg secretion is greater than the additive effect.

[0197] Table 5: Total IFNg secretion in co-culture experiments of FluM1 viral peptide-specific T cells and HCC2935 cells loaded with FluM1 peptide in the presence of anti-CEACAM6 and anti-PD-1 antibodies (as single agents or in combination) (n=7)

[0198]

[0199] Description table:HCC2935 lung cancer cells (HCC) were pulsed with 0.2 μg / ml FluM1 peptide to stimulate viral peptide-specific T cells (TC) in co-culture. 30 μg / ml of antibody was applied. For combination treatment (n=7), 1 μg / ml of TPP-3310 was added. The concentration of secreted IFN-γ was determined by ELISA, and the data are isotype-corrected values ​​and expressed in pg / ml. TPP-3310, aCEACAM6; TPP-2596, aPD-1.

[0200] T-test of the means, p-value (<0.05): aPD-1 vs. CEACAM6, p=0.13; aPD-L1 vs. combination, p=0.0034; aCEACAM6 vs. combination, p=0.0011 Sequence Listing <110> Bayer AG <120> Drug combinations used to treat cancer <130> BHC181044 <160> 70 <170> PatentIn version 3.5 <210> 1 <211> 113 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 1 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 <210> 2 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 2 Asn Ser Gly Met His 1 5 <210> 3 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 3 Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 4 <211> 4 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 4 Asn Asp Asp Tyr 1 <210> 5 <211> 107 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 5 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 100 105 <210> 6 <211> 11 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 6 Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala 1 5 10 <210> 7 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 7 Asp Ala Ser Asn Arg Ala Thr 1 5 <210> 8 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 8 Gln Gln Ser Ser Asn Trp Pro Arg Thr 1 5 <210> 9 <211> 440 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 9 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 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 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> 10 <211> 214 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 10 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 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> 11 <211> 118 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 11 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Tyr 20 25 30 Gly Ile Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala His Ile Trp Trp Asn Asp Asn Lys Tyr Tyr Ser Thr Ser 50 55 60 Leu Lys Thr Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ile Ser Leu Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser 115 <210> 12 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Antibody Sequence <400> 12 Thr Tyr Gly Ile Gly Val Gly 1 5 <210> 13 <211> 16 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 13 His Ile Trp Trp Asn Asp Asn Lys Tyr Tyr Ser Thr Ser Leu Lys Thr 1 5 10 15 <210> 14 <211> 8 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 14 Ile Ser Leu Pro Tyr Phe Asp Tyr 1 5 <210> 15 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 15 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Asn Arg Tyr 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 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 16 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 16 Lys Ala Ser Gln Asn Val Gly Thr Ala Val Ala 1 5 10 <210> 17 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 17 Ser Ala Ser Asn Arg Tyr Thr 1 5 <210> 18 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 18 Gln Gln Tyr Ser Ser Tyr Pro Leu Thr 1 5 <210> 19 <211> 443 <212> PRT <213> Artificial sequence <220> <223> Antibody sequence <400> 19 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Tyr 20 25 30 Gly Ile Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala His Ile Trp Trp Asn Asp Asn Lys Tyr Tyr Ser Thr Ser 50 55 60 Leu Lys Thr Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ile Ser Leu Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys 210 215 220 Pro Pro Cys Pro Ala Pro Pro Val Ala 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 His 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 Phe Arg Val Val Ser Val Leu Thr 290 295 300 Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 305 310 315 320 Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr 325 330 335 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 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 Met Leu Asp Ser Asp Gly Ser 385 390 395 400 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 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 Pro Gly 435 440 <210> 20 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> antibody sequence <400> 20 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Asn Arg Tyr 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 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser 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 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> 21 <211> 118 <212> PRT <213> Artificial sequence <220> <223> Antibody sequence <400> 21 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 Asp Ser 20 25 30 Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala 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 His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala 115 <210> twenty two <211> 5 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> twenty two Asp Ser Trp Ile His 1 5 <210> twenty three <211> 17 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> twenty three Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> twenty four <211> 11 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> twenty four Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr 1 5 10 <210> 25 <211> 107 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 25 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 Asp Val Ser Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Phe Leu Tyr 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 Leu Tyr His Pro Ala 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 26 <211> 11 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 26 Arg Ala Ser Gln Asp Val Ser Thr Ala Val Ala 1 5 10 <210> 27 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 27 Serve me, I'll be there soon 1 5 <210> 28 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 28 Gln Gln Tyr Leu Tyr His Pro Ala Thr 1 5 <210> 29 <211> 445 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 29 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 Asp Ser 20 25 30 Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala 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 His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ala 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 Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val 210 215 220 Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala 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 Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val 290 295 300 Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Thr 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 Met 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 435 440 445 <210> 30 <211> 214 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 30 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 Asp Val Ser Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Phe Leu Tyr 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 Leu Tyr His Pro Ala 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 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> 31 <211> 120 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 31 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 115 120 <210> 32 <211> 5 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 32 Asn Tyr Tyr Met Tyr 1 5 <210> 33 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 33 Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe Lys 1 5 10 15 Asn <210> 34 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 34 Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr 1 5 10 <210> 35 <211> 111 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 35 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 100 105 110 <210> 36 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 36 Arg Ala Ser Lys Gly Val Ser Thr Ser Gly Tyr Ser Tyr Leu His 1 5 10 15 <210> 37 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 37 Leu Ala Ser Tyr Leu Glu Ser 1 5 <210> 38 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 38 Gln His Ser Arg Asp Leu Pro Leu Thr 1 5 <210> 39 <211> 447 <212> PRT <213> Artificial Sequence <220> <223> Antibody Sequence <400> 39 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> 40 <211> 218 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 40 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> 41 <211> 118 <212> PRT <213> Synthetic Sequence <220> <223> Antibody Sequence <400> 41 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 Asp Ser 20 25 30 Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala 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 His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <210> 42 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 42 Asp Ser Trp Ile His 1 5 <210> 43 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 43 Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 44 <211> 9 <212> PRT <213> Artificial sequence <220> <223> antibody sequence <400> 44 Arg His Trp Pro Gly Gly Phe Asp Tyr 1 5 <210> 45 <211> 107 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 45 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 Asp Val Ser Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Phe Leu Tyr 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 Leu Tyr His Pro Ala 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 46 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 46 Arg Ala Ser Gln Asp Val Ser Thr Ala Val Ala 1 5 10 <210> 47 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 47 Serve me, I'll be there soon 1 5 <210> 48 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 48 Gln Gln Tyr Leu Tyr His Pro Ala Thr 1 5 <210> 49 <211> 448 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 49 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 Asp Ser 20 25 30 Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala 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 His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 50 <211> 214 <212> PRT <213> Synthetic Sequence <220> <223> Antibody Sequence <400> 50 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 Asp Val Ser Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Phe Leu Tyr 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 Leu Tyr His Pro Ala 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 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> 51 <211> 120 <212> PRT <213> Artificial Sequence <220> <223> Antibody Sequence <400> 51 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 Thr 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 Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 52 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 52 Ser Tyr Ile Met 1 5 <210> 53 <211> 17 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 53 Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr Val Lys 1 5 10 15 Gly <210> 54 <211> 11 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 54 Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr 1 5 10 <210> 55 <211> 110 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 55 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> 56 <211> 14 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 56 Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser 1 5 10 <210> 57 <211> 7 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 57 Asp Val Ser Asn Arg Pro Ser 1 5 <210> 58 <211> 10 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 58 Ser Ser Tyr Thr Ser Ser Ser Thr Arg Val 1 5 10 <210> 59 <211> 450 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 59 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 Thr 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 Ile Lys Leu Gly Thr Val Thr Thr Val 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 Ser Ser Lys Ser Thr Ser Gly Gly 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 Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Light 450 <210> 60 <211> 216 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 60 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 Gly Gln 100 105 110 Pro Lys Ala Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser Glu Glu 115 120 125 Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr 130 135 140 Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro Val Lys 145 150 155 160 Ala Gly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn Lys Tyr 165 170 175 Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His 180 185 190 Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys 195 200 205 Thr Val Ala Pro Thr Glu Cys Ser 210 215 <210> 61 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> Antibody Sequence <400> 61 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Gly Gly Trp Phe Gly Glu Leu Ala Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 62 <211> 5 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 62 Arg Tyr Trp Met Ser 1 5 <210> 63 <211> 17 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 63 Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val Lys 1 5 10 15 Gly <210> 64 <211> 12 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 64 Glu Gly Gly Trp Phe Gly Glu Leu Ala Phe Asp Tyr 1 5 10 <210> 65 <211> 108 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 65 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Arg Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Asp Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Leu Pro 85 90 95 Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 66 <211> 12 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 66 Arg Ala Ser Gln Arg Val Ser Ser Ser Tyr Leu Ala 1 5 10 <210> 67 <211> 7 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 67 Asp Ala Ser Ser Arg Ala Thr 1 5 <210> 68 <211> 9 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 68 Gln Gln Tyr Gly Ser Leu Pro Trp Thr 1 5 <210> 69 <211> 451 <212> PRT <213> Artificial sequence <220> <223> Antibody sequences <400> 69 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30 Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Gly Gly Trp Phe Gly Glu Leu Ala Phe Asp Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 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 Phe Glu 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 Ser 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 Light 450 <210> 70 <211> 215 <212> PRT <213> artificial sequence <220> <223> antibody sequence <400> 70 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Arg Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Asp Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Leu Pro 85 90 95 Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro 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

Claims

1. A combination of an anti-CEACAM6 antibody and an anti-PD-1 antibody or an anti-PD-L1 antibody, wherein: The anti-CEACAM6 antibody is used for treating cancer simultaneously, separately or sequentially in combination with the anti-PD-1 antibody or the anti-PD-L1 antibody, wherein the anti-CEACAM6 antibody comprises H-CDR1 as shown in the amino acid sequence of SEQ ID NO: 12, H-CDR2 as shown in the amino acid sequence of SEQ ID NO: 13, H-CDR3 as shown in the amino acid sequence of SEQ ID NO: 14, L-CDR1 as shown in the amino acid sequence of SEQ ID NO: 16, L-CDR2 as shown in the amino acid sequence of SEQ ID NO: 17 and L-CDR3 as shown in the amino acid sequence of SEQ ID NO:

18. 2 . The combination according to claim 1 , wherein the anti-CEACAM6 antibody comprises a variable heavy chain sequence of SEQ ID NO: 11 and a variable light chain sequence of SEQ ID NO:

15.

3. The combination of claim 1, wherein the anti-CEACAM6 antibody comprises a heavy chain region of SEQ ID NO: 19 and a light chain region of SEQ ID NO:

20.

4. The combination according to any one of claims 1-3, wherein the anti-PD-1 antibody is nivolumab or pembrolizumab, and the anti-PD-L1 antibody is atezolizumab, avelumab or durvalumab.

5. A combination according to any one of claims 1 to 3, The anti-PD-1 antibody comprises: (i) H-CDR1 as shown in the amino acid sequence of SEQ ID NO:2, H-CDR2 as shown in the amino acid sequence of SEQ ID NO:3, H-CDR3 as shown in the amino acid sequence of SEQ ID NO:4, L-CDR1 as shown in the amino acid sequence of SEQ ID NO:6, L-CDR2 as shown in the amino acid sequence of SEQ ID NO:7, and L-CDR3 as shown in the amino acid sequence of SEQ ID NO:8, or (ii) H-CDR1 as shown in the amino acid sequence of SEQ ID NO:32, H-CDR2 as shown in the amino acid sequence of SEQ ID NO:33, H-CDR3 as shown in the amino acid sequence of SEQ ID NO:34, L-CDR1 as shown in the amino acid sequence of SEQ ID NO:36, L-CDR2 as shown in the amino acid sequence of SEQ ID NO:37, and L-CDR3 as shown in the amino acid sequence of SEQ ID NO:38; and wherein the anti-PD-L1 antibody comprises: (iii) H-CDR1 as shown in the amino acid sequence of SEQ ID NO:42, H-CDR2 as shown in the amino acid sequence of SEQ ID NO:43, H-CDR3 as shown in the amino acid sequence of SEQ ID NO:44, L-CDR1 as shown in the amino acid sequence of SEQ ID NO:46, L-CDR2 as shown in the amino acid sequence of SEQ ID NO:47, and L-CDR3 as shown in the amino acid sequence of SEQ ID NO:48, or (iv) H-CDR1 as shown in the amino acid sequence of SEQ ID NO:52, H-CDR2 as shown in the amino acid sequence of SEQ ID NO:53, H-CDR3 as shown in the amino acid sequence of SEQ ID NO:54, L-CDR1 as shown in the amino acid sequence of SEQ ID NO:56, L-CDR2 as shown in the amino acid sequence of SEQ ID NO:57, and L-CDR3 as shown in the amino acid sequence of SEQ ID NO:58, or (v) H-CDR1 as shown in the amino acid sequence of SEQ ID NO:62, H-CDR2 as shown in the amino acid sequence of SEQ ID NO:63, H-CDR3 as shown in the amino acid sequence of SEQ ID NO:64, L-CDR1 as shown in the amino acid sequence of SEQ ID NO:66, L-CDR2 as shown in the amino acid sequence of SEQ ID NO:67, and L-CDR3 as shown in the amino acid sequence of SEQ ID NO:

68.

6. A combination according to any one of claims 1 to 3, The anti-PD-1 antibody comprises: (i) the variable heavy chain sequence (VH) of SEQ ID NO: 1 and the variable light chain sequence (VL) of SEQ ID NO: 5, or (ii) the variable heavy chain sequence (VH) of SEQ ID NO:31 and the variable light chain sequence (VL) of SEQ ID NO:35, and wherein the anti-PD-L1 antibody comprises: (iii) the variable heavy chain sequence (VH) of SEQ ID NO:41 and the variable light chain sequence (VL) of SEQ ID NO:45, or (iv) the variable heavy chain sequence (VH) of SEQ ID NO:51 and the variable light chain sequence (VL) of SEQ ID NO:55, or (v) the variable heavy chain sequence (VH) of SEQ ID NO:61 and the variable light chain sequence (VL) of SEQ ID NO:

65.

7. A combination according to any one of claims 1 to 3, The anti-PD-1 antibody comprises: (i) the heavy chain region (HC) of SEQ ID NO:9 and the light chain region (LC) of SEQ ID NO:10, or (ii) the heavy chain region (HC) of SEQ ID NO:39 and the light chain region (LC) of SEQ ID NO:40, and wherein the anti-PD-L1 antibody comprises: (iii) the heavy chain region (HC) of SEQ ID NO:49 and the light chain region (LC) of SEQ ID NO:50, or (iv) the heavy chain region (HC) of SEQ ID NO:59 and the light chain region (LC) of SEQ ID NO:60, or (v) the heavy chain region (HC) of SEQ ID NO:69 and the light chain region (LC) of SEQ ID NO:

70.

8. The combination according to any one of claims 1-7, wherein the cancer is lung cancer, ovarian cancer, mesothelioma, pancreatic cancer, gastric cancer, colorectal cancer, head and neck cancer, bladder cancer, bile duct cancer, breast cancer, cervical cancer or esophageal cancer.

9. The combination according to claim 8, wherein the cancer is non-small cell lung cancer.

10. The combination according to any one of claims 1-9, wherein at least one of the anti-CEACAM6 antibody, anti-PD-1 antibody or anti-PD-L1 antibody is administered simultaneously, separately or sequentially in combination with one or more pharmaceutical agents.

11. Use of a combination according to any one of claims 1 to 10 for the preparation of a medicament for the treatment of cancer, wherein the cancer is lung cancer, ovarian cancer, mesothelioma, pancreatic cancer, gastric cancer, colorectal cancer, head and neck cancer, bladder cancer, bile duct cancer, breast cancer, cervical cancer or esophageal cancer.

12. The use according to claim 11, wherein the cancer is non-small cell lung cancer.

13. The use according to claim 11, wherein at least one of the anti-CEACAM6 antibody, anti-PD-1 antibody or anti-PD-L1 antibody is administered simultaneously, separately or sequentially in combination with one or more pharmaceutical agents.

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