Anti-cancer combination therapy targeting CD38 and TGF-β

Abstract

The present disclosure relates to a combination cancer therapy targeting CD38 and TGF-β, which uses antibodies specific for these targets. Compositions useful in the therapy are also provided.

Description

[0001] Cross - reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application 62 / 696,198, filed on July 10, 2018. The disclosure of this application is hereby incorporated by reference in its entirety.

[0003] Sequence Listing

[0004] This application contains a Sequence Listing that has been electronically submitted in ASCII format and is hereby incorporated by reference in its entirety. The electronic copy of the Sequence Listing created on July 9, 2019, is named 022548_WO025_SL.txt and is 52,428 bytes in size. Background of the Invention

[0005] Transforming growth factor - β (TGF - β) is a cytokine that regulates multiple biological processes such as extracellular matrix formation, wound healing, embryonic development, skeletal development, hematopoiesis, immune and inflammatory responses, and malignant transformation. Dysregulation of TGF - β leads to pathological conditions such as birth defects, cancer, chronic inflammation, and autoimmune and fibrotic diseases.

[0006] TGF - β has three known isoforms: TGF - β1, 2, and 3. These three isoforms are pleiotropic in their functions and are expressed in different patterns between cell and tissue types. They have similar in vitro activities, but single knockouts in specific cell types indicate that although they share the ability to bind to the same receptors, they have different roles in vivo (Akhurst et al., Nat Rev Drug Discov 11(10):790 - 811(2012)).

[0007] After TGF - β binds to TGFβRII, the constitutive kinase activity of the receptor phosphorylates and activates TGFβRI, thereby phosphorylating SMAD2 / 3, which allows SMAD2 / 3 to associate with SMAD4 and then localize to the nucleus and initiate transcription of TGF - β responsive genes. Ibid. In addition to this classical signaling cascade, non - classical pathways also transmit signals through other factors including p38 MAPK, PI3K, AKT, JUN, JNK, and NF - κB. TGF - β signaling is also regulated by other pathways including WNT, Hedgehog, Notch, INF, TNF, and RAS. Thus, the end result of TGF - β signaling is the crosstalk of all these signaling pathways, which integrates the state and environment of the cell. Ibid.

[0008] CD38 is a 45kD type II transmembrane glycoprotein with a long C-terminal extracellular domain and a short N-terminal cytoplasmic domain. The CD38 protein is a bifunctional extracellular enzyme that catalyzes the conversion of NAD + to cyclic ADP-ribose (cADPR) and the hydrolysis of cADPR to ADP-ribose. During ontogeny, CD38 appears on CD34-positive committed stem cells and lineage-committed progenitors of lymphocytes, erythroid cells, and myeloid cells. CD38 expression persists mainly in the lymphoid lineage and has different expression levels at different stages of T cell and B cell development.

[0009] CD38 is upregulated in many hematopoietic malignancies and cell lines derived from various hematopoietic malignancies, including non-Hodgkin lymphoma (NHL), Burkitt lymphoma (BL), multiple myeloma (MM), B chronic lymphocytic leukemia (B-CLL), B and T acute lymphocytic leukemia (ALL), T cell lymphoma (TCL), acute myeloid leukemia (AML), hairy cell leukemia (HCL), Hodgkin lymphoma (HL), and chronic myeloid leukemia (CML). On the other hand, the most primitive pluripotent stem cells of the hematopoietic system are CD38-negative. The CD38 expression in hematopoietic malignancies and its correlation with disease progression make CD38 an attractive target for antibody therapy. SUMMARY OF THE INVENTION

[0010] The present invention provides a combination therapy targeting CD38 and TGF-β. The inventors have found that anti-TGF-β antibodies can block the ability of TGF-β to attenuate the anti-tumor effects of anti-CD38 antibodies (e.g., TGF-β can inhibit the NK cell-mediated ADCC of anti-CD38 antibodies). Compared with currently available cancer treatments, including antibody therapy, the combination therapy provided herein can offer excellent clinical efficacy.

[0011] Accordingly, the present disclosure provides a method for enhancing the efficacy of an agent that specifically binds to CD38 with an agent that specifically binds to TGF-β. In some embodiments, the present disclosure provides a method for treating cancer in a patient (e.g., a human patient) by administering to the patient an agent that specifically binds to human CD38 and an agent that specifically binds to human TGF-β. In some embodiments, the agent that specifically binds to human CD38 is an anti-CD38 antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-CD38 antibody is capable of killing CD38-positive cells by apoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC), and / or complement-dependent cytotoxicity (CDC), wherein killing of the CD38-positive cells by apoptosis can occur in the absence of stromal cells or stromal-derived cytokines. In some embodiments, the agent that specifically binds to human TGF-β is a pan-specific anti-TGF-β antibody or an antigen-binding fragment thereof.

[0012] In some embodiments, the present disclosure provides a method for treating cancer in a human patient in need thereof, the method comprising administering to the patient an anti-CD38 antibody and an anti-TGF-β antibody or an antigen fragment thereof.

[0013] In some embodiments, the anti-CD38 antibody:

[0014] a) has HCDR1-3 and LCDR1-3 having amino acid sequences comprising SEQ ID NOs: 15-20, respectively;

[0015] b) has a heavy chain variable domain and a light chain variable domain having amino acid sequences comprising SEQ ID NOs: 13 and 14, respectively; or

[0016] c) has a heavy chain having an amino acid sequence comprising SEQ ID NO: 11 and a light chain having an amino acid sequence comprising SEQ ID NO: 12.

[0017] In some embodiments, the anti-TGF-β antibody:

[0018] a) has HCDR1-3 and LCDR1-3 having amino acid sequences comprising SEQ ID NOs: 5-10, respectively;

[0019] b) has a heavy chain variable domain and a light chain variable domain having amino acid sequences comprising SEQ ID NOs: 3 and 4, respectively; or

[0020] c) has a heavy chain having an amino acid sequence comprising SEQ ID NO: 1 and a light chain having an amino acid sequence comprising SEQ ID NO: 2.

[0021] In some embodiments, the anti-CD38 antibody has heavy chain complementarity-determining region 1 (HCDR1), HCDR2, HCDR3, light chain complementarity-determining region 1 (LCDR1), LCDR2, and LCDR3 that respectively comprise the amino acid sequences of SEQ ID NOs: 15-20, and the anti-TGF-β antibody or antigen-binding fragment thereof has HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 that respectively comprise the amino acid sequences of SEQ ID NOs: 5-10. In certain embodiments, the anti-CD38 antibody comprises a heavy chain variable domain (V H ) and a light chain variable domain (V L ) that respectively comprise the amino acid sequences of SEQ ID NOs: 13 and 14; and the anti-TGF-β antibody comprises V H and V L that respectively comprise the amino acid sequences of SEQ ID NOs: 3 and 4. In specific embodiments, the anti-CD38 antibody has a heavy chain (HC) and a light chain (LC) that respectively comprise the amino acid sequences of SEQ ID NOs: 11 and 12; and the anti-TGF-β antibody has a heavy chain and a light chain that respectively comprise the amino acid sequences of SEQ ID NOs: 1 and 2.

[0022] In any of the embodiments provided herein, the anti-CD38 antibody can comprise a human IgG1 Fc region, and the anti-TGF-β antibody can comprise a human IgG4 Fc region.

[0023] Also provided herein is a method of treating multiple myeloma in a human patient in need thereof, the method comprising administering to the patient an anti-CD38 antibody and an anti-TGF-β antibody, the anti-CD38 antibody comprising the heavy chain variable domain amino acid sequence of SEQ ID NO: 13 and the light chain variable domain amino acid sequence of SEQ ID NO: 14, and the anti-TGF-β antibody comprising the heavy chain variable domain amino acid sequence of SEQ ID NO: 3 and the light chain variable domain amino acid sequence of SEQ ID NO: 4.

[0024] Also provided herein is a method of treating multiple myeloma in a human patient in need thereof, the method comprising administering to the patient an anti-CD38 antibody and an anti-TGF-β antibody, the anti-CD38 antibody comprising the heavy chain amino acid sequence of SEQ ID NO: 11 and the light chain amino acid sequence of SEQ ID NO: 12, and the anti-TGF-β antibody comprising the heavy chain amino acid sequence of SEQ ID NO: 1 and the light chain amino acid sequence of SEQ ID NO: 2.

[0025] In any of the embodiments provided herein, the anti-CD38 antibody and the anti-TGF-β antibody or fragment can be administered sequentially to the patient.

[0026] Also provided herein is an anti-CD38 antibody for use in combination with an anti-TGF-β antibody for treating cancer in a human patient in need thereof, and the use of an anti-CD38 antibody in combination with an anti-TGF-β antibody for the manufacture of a medicament for treating cancer in a human patient in need thereof. In some embodiments, the anti-CD38 antibody

[0027] a) has HCDR1-3 and LCDR1-3 respectively comprising the amino acid sequences of SEQ ID NOs: 15-20;

[0028] b) has a heavy chain variable domain and a light chain variable domain respectively comprising the amino acid sequences of SEQ ID NOs: 13 and 14; or

[0029] c) has a heavy chain comprising the amino acid sequence of SEQ ID NO: 11 and a light chain comprising the amino acid sequence of SEQ ID NO: 12.

[0030] Also provided herein is an anti-TGF-β antibody for use in combination with an anti-CD38 antibody for treating cancer in a human patient in need thereof, and the use of an anti-TGF-β antibody in combination with an anti-CD38 antibody for the manufacture of a medicament for treating cancer in a human patient in need thereof. In some embodiments, the anti-TGF-β antibody

[0031] a) has HCDR1-3 and LCDR1-3 respectively comprising the amino acid sequences of SEQ ID NOs: 5-10;

[0032] b) has a heavy chain variable domain and a light chain variable domain respectively comprising the amino acid sequences of SEQ ID NOs: 3 and 4; or

[0033] c) has a heavy chain comprising the amino acid sequence of SEQ ID NO: 1 and a light chain comprising the amino acid sequence of SEQ ID NO: 2.

[0034] In some embodiments of the methods, antibodies for the uses, and uses of the antibodies described herein, the cancer is CD38 positive.

[0035] In some embodiments of the methods, antibodies for the uses, and uses of antibodies described herein, the cancer is selected from multiple myeloma, non-Hodgkin lymphoma, Hodgkin lymphoma, diffuse large B-cell lymphoma, peripheral T-cell lymphoma, hairy cell leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, acute lymphocytic leukemia, melanoma, glioblastoma, lung cancer, cutaneous squamous cell carcinoma, colorectal cancer, breast cancer, ovarian cancer, head and neck cancer, hepatocellular carcinoma, urothelial carcinoma, and renal cell carcinoma.

[0036] In some embodiments of the methods, antibodies for the uses, and uses of antibodies described herein, the cancer is a hematological malignancy.

[0037] In some embodiments of the methods, antibodies for the uses, and uses of antibodies described herein, the cancer is multiple myeloma. In certain embodiments, in patients with multiple myeloma, the treatment described herein results in less bone destruction and / or enhanced bone formation compared to treatment with an anti-CD38 antibody alone, leading to bone remodeling and / or bone healing.

[0038] Thus, in some embodiments, provided herein is a method for reducing bone destruction in a human patient with multiple myeloma, the method comprising administering to the patient an agent that specifically binds to human CD38 as described herein (e.g., an anti-CD38 antibody) and an agent that specifically binds to human TGF-β as described herein (e.g., an anti-TGF-β antibody). Also provided herein is an anti-TGF-β antibody as described herein for use in combination with an anti-CD38 antibody for reducing bone destruction in a human patient with multiple myeloma, and the use of an anti-TGF-β antibody in combination with an anti-CD38 antibody for the manufacture of a medicament for reducing bone destruction in a human patient with multiple myeloma.

[0039] Also provided herein is a method for enhancing bone formation in a patient with multiple myeloma, leading to bone remodeling and / or bone healing, the method comprising administering to the patient an agent that specifically binds to human CD38 as described herein (e.g., an anti-CD38 antibody) and an agent that specifically binds to human TGF-β as described herein (e.g., an anti-TGF-β antibody). Also provided herein is an anti-TGF-β antibody as described herein for use in combination with an anti-CD38 antibody for enhancing bone formation in a human patient with multiple myeloma, and the use of an anti-TGF-β antibody in combination with an anti-CD38 antibody for the manufacture of a medicament for enhancing bone formation in a human patient with multiple myeloma.

[0040] In some embodiments of the methods, antibodies for the uses, and uses of antibodies described herein, the cancer is refractory to treatment with Ab2 or with daratumumab, or is refractory to both treatments. In some embodiments, the patient has relapsed or refractory multiple myeloma and has received at least one prior therapy or at least two prior therapies.

[0041] Also provided herein are articles comprising an anti-CD38 antibody and an anti-TGF-β antibody, wherein the article is suitable for treating cancer in a patient, for example, in the treatment methods described herein. In some embodiments, the anti-CD38 antibody

[0042] a) has HCDR1-3 and LCDR1-3 respectively comprising the amino acid sequences of SEQ ID NOs: 15-20;

[0043] b) has a heavy chain variable domain and a light chain variable domain respectively comprising the amino acid sequences of SEQ ID NOs: 13 and 14; or

[0044] c) has a heavy chain comprising the amino acid sequence of SEQ ID NO: 11 and a light chain comprising the amino acid sequence of SEQ ID NO: 12; and

[0045] the anti-TGF-β antibody

[0046] a) has HCDR1-3 and LCDR1-3 respectively comprising the amino acid sequences of SEQ ID NOs: 5-10;

[0047] b) has a heavy chain variable domain and a light chain variable domain respectively comprising the amino acid sequences of SEQ ID NOs: 3 and 4; or

[0048] c) has a heavy chain comprising the amino acid sequence of SEQ ID NO: 1 and a light chain comprising the amino acid sequence of SEQ ID NO: 2. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] FIG. 1 is a graph showing TGF-β released by multiple myeloma cell lines at 1, 2, or 3 days as quantified by ELISA.

[0050] FIG. 2 is a graph showing the calcein fluorescence levels (lysis %) from MOLP8 target cells, which were incubated with Ab2 at concentrations of 0, 0.001, 0.1, or 1 μg / mL and lysed for 1 hour by human NK cells incubated overnight, 24 hours, 48 hours, or 72 hours with 10 ng / mL TGF-β.

[0051] Figure 3 is a graph showing the calcein fluorescence levels (percent lysis) from MOLP8 target cells that were incubated with Ab2 or IgG1 control at concentrations of 0, 0.001, 0.01, 0.1, or 1 μg / mL and lysed for 1 hour by human NK cells incubated with 10 ng / mL TGF-β and 50 μg / mL Ab1 for 90 hours.

[0052] Figure 4 is a graph showing the effect of 0.1, 1, and 10 ng / mL TGF-β on Ab2-mediated ADCC of NCI-H929 cells.

[0053] Figure 5 is a graph showing Ab2-mediated ADCC of NCI-H929 cells treated with 100 μg / mL IgG4, 100 μg / mL Ab1, 10 ng / mL TGF-β, TGF-β + IgG4, or TGF-β + Ab1. * = p < 0.05, compared to NT; ** = p < 0.005, compared to NT.

[0054] Figure 6 is a graph showing Ab2-mediated ADCC of NCI-H929 cells treated with 200 μg / mL IgG4, 200 μg / mL Ab1, 10 ng / mL TGF-β, TGF-β + IgG4, or TGF-β + Ab1. * = p < 0.05, compared to NT; ** = p < 0.005, compared to NT.

[0055] Figure 7 is a graph showing the calcein fluorescence levels (percent lysis) from K562 target cells that were lysed by the cytotoxic activity of human NK cells incubated with JJN3 cells releasing endogenous TGF-β and 100 μg / mL Ab1 or isotype control for 90 hours.

[0056] Figure 8 is a graph showing Ab2-mediated ADCC of RPMI8226 target cells that were incubated with Ab2 at concentrations of 0, 0.01, or 0.1 μg / mL or with a control Ab2 mutant at a concentration of 0.1 μg / mL and lysed for 1 hour by human NK cells incubated with JJN3 cells releasing endogenous TGF-β and 100 μg / mL Ab1 or isotype control for 90 hours. Detailed Description

[0057] The present disclosure provides novel combination therapies targeting human TGF-β and human CD38, such as by using antibodies that bind to these targets. The combination therapies can be used to treat conditions such as cancer. Unless otherwise specified, "TGF-β" herein refers to human TGF-β. The polypeptide sequences of the three isoforms of human TGF-β (TGF-β1, TGF-β2, and TGF-β3) are available under SwissProt accession numbers P01137, P08112, and P10600, respectively, and are shown herein as SEQ ID NO: 21-23. Unless otherwise specified, "CD38" herein refers to human CD38. The human CD38 polypeptide sequence is available under Genbank accession number NP_001766 and is shown herein as SEQ ID NO: 24.

[0058] As used herein, the term "antibody" (Ab) or "immunoglobulin" (Ig) refers to a tetrameric protein comprising two heavy (H) chains (about 50-70 kDa) and two light (L) chains (about 25 kDa) that are interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable domain (V H ) and a heavy chain constant region (C H ). Each light chain consists of a light chain variable domain (V L ) and a light chain constant region (C L ). The V H and V L domains can be further subdivided into regions of high variability called "complementary determining regions" (CDRs), interspersed with more conserved regions called "framework regions" (FRs). Each V H or V L is composed of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The assignment of amino acids in each region can be based on the definition (Lefranc et al., Dev Comp Immunol 27(1):55-77 (2003)); or the following definition: Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Maryland (1987 and 1991)); or Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987); or Chothia et al., Nature 342:878-883 (1989).

[0059] The term "affinity" refers to a measure of the attraction between an antigen and an antibody. The inherent attraction of an antibody for an antigen is usually expressed as the binding affinity equilibrium constant (K D ) of a particular antibody-antigen interaction. As determined, for example, by surface plasmon resonance or biolayer interferometry, when K D is less than 10 -7 M (such as less than 10 -8 M (e.g., 1 - 9 nM)), the antibody is said to specifically bind to the antigen.

[0060] The term "k off " refers to the dissociation rate constant of a particular antibody-antigen interaction. The k off dissociation rate constant can be measured, for example, by biolayer interferometry.

[0061] As used herein, the term "epitope" refers to the part (determinant) of an antigen that specifically binds to an antibody. Epitope determinants are usually composed of the chemically reactive surface groups of a molecule (such as amino acids or carbohydrates or sugar side chains) and usually have specific three-dimensional structural features, as well as specific charge characteristics. Epitopes can be "linear" or "conformational". In a linear epitope, all the interaction points between a protein (e.g., an antigen) and an interacting molecule (e.g., an antibody) are linearly present along the primary amino acid sequence of the protein. In a conformational epitope, the interaction points span amino acid residues on the protein that are separated from each other in the primary amino acid sequence. Once the desired epitope on an antigen is identified, antibodies specific for that epitope can be generated using techniques well known in the art. For example, antibodies specific for a linear epitope can be generated, for example, by immunizing an animal with a peptide having the amino acid residues of the linear epitope. Antibodies specific for a conformational epitope can be generated, for example, by immunizing an animal with a microdomain containing the relevant amino acid residues of the conformational epitope. Antibodies specific for a particular epitope can also be generated, for example, by immunizing an animal with the target molecule of interest or a relevant portion thereof and then screening for binding to the epitope.

[0062] Antibodies can be determined to bind the same epitope as an antibody described herein or to compete for binding with an antibody described herein by using methods known in the art, including but not limited to competitive assays, epitope binning, and alanine scanning. In some embodiments, the test antibody and an antibody described herein bind to at least one common residue (e.g., at least two, three, four, five, six, seven, eight, nine, or ten residues) on the target protein (i.e., TGF-β or CD38). In further embodiments, the contacting residues on the target protein are identical between the test antibody and an antibody described herein. In one embodiment, an antibody described herein is bound to the target protein under saturating conditions, and then the ability of the test antibody to bind to the target protein is measured. If the test antibody can bind the target protein simultaneously with the reference antibody, the test antibody binds a different epitope from the reference antibody. However, if the test antibody cannot bind the target protein simultaneously, the test antibody binds the same epitope, an overlapping epitope, or an epitope very close to the epitope bound by an antibody described herein. The experiment can be performed using, for example, ELISA, RIA, BIACORE TM , SPR, biolayer interferometry, or flow cytometry. To test whether one antibody cross-competes with another, the above-described competitive methods can be used in both directions, i.e., to determine whether a known antibody blocks the test antibody and vice versa.

[0063] The term "humanized" refers to the fact that an antibody that is wholly or partially of non-human origin (e.g., a murine antibody obtained by immunizing a mouse with an antigen of interest, or a chimeric antibody based on such a murine antibody) can have certain amino acids replaced, particularly in the framework and constant regions of the heavy and light chains, to avoid or minimize the immune response in humans. Although it is not possible to precisely predict the immunogenicity of a particular antibody and thus the human anti-antibody response, non-human antibodies tend to be more immunogenic in humans than human antibodies. Chimeric antibodies in which the foreign (e.g., rodent) constant regions have been replaced with sequences of human origin have been shown to generally have lower immunogenicity than antibodies of completely foreign origin, and therapeutic antibodies tend to be humanized or fully human antibodies. Chimeric antibodies or other antibodies of non-human origin can thus be humanized to reduce the risk of human anti-antibody responses.

[0064] For chimeric antibodies, humanization typically involves modification of the framework regions of the variable domain sequences. Amino acid residues that are part of the complementarity determining regions (CDRs) are generally not altered with respect to humanization, but in some cases it may be desirable to alter individual CDR amino acid residues, for example to remove glycosylation sites, deamidation sites, aspartic acid isomerization sites or unwanted cysteine or methionine residues. N-linked glycosylation occurs by attachment of an oligosaccharide chain to an asparagine residue in the tripeptide sequence Asn-X-Ser or Asn-X-Thr, where X can be any amino acid other than Pro. Removal of N-glycosylation sites can be achieved by mutating the Asn or Ser / Thr residue to a different residue, preferably by conservative substitution. Deamidation of asparagine and glutamine residues can occur depending on factors such as pH and surface exposure. Asparagine residues are particularly prone to deamidation, mainly when present in the Asn-Gly sequence, and to a lesser extent in other dipeptide sequences such as Asn-Ala. When such deamidation sites, particularly Asn-Gly, are present in the CDR sequence, it may therefore be desirable to remove the site, usually by conservative substitution to remove one of the residues involved.

[0065] Many methods for humanizing antibody sequences are known in the art; see, for example, the review by Almagro and Fransson, Front Biosci. 13:1619-1633 (2008). One commonly used method is CDR grafting, which for example for a murine-derived chimeric antibody involves identifying the human germline gene counterparts of the murine variable domain genes and grafting the murine CDR sequences into that framework. The specificity of the interaction of an antibody with its target antigen lies mainly in the amino acid residues in the six CDRs of the heavy and light chains. Thus, the amino acid sequence within the CDRs is much more variable between individual antibodies than the sequences outside the CDRs. Since the CDR sequences are responsible for most antibody-antigen interactions, recombinant antibodies can be expressed that mimic the properties of naturally occurring specific antibodies, or more generally any specific antibody with a given amino acid sequence, for example by constructing an expression vector that expresses CDR sequences from a specific antibody grafted into the framework sequences from different antibodies. Thus, non-human antibodies can be "humanized" and still substantially retain the binding specificity and affinity of the original antibody. CDR grafting can be based on the Kabat CDR definitions, but more recent publications (Magdelaine-Beuzelin et al., Crit Rev Oncol Hematol. 64:210-225 (2007)) have proposed definitions (the international ImMunoGeneTics information www.imgt.org) can improve the results of humanization (see Lefranc et al., Dev. Comp Immunol. 27:55-77 (2003)).

[0066] In some cases, compared to the parental antibody from which the CDR is obtained, CDR grafting may reduce the binding specificity and affinity of the non-human antibody with the CDR grafted, and thus reduce its biological activity. Back mutations (sometimes called "framework repair") can be introduced at selected positions (usually in the framework regions) in the antibody with the CDR grafted to re-establish the binding specificity and affinity of the parental antibody. Information available in the literature and antibody databases can be used to identify the positions of possible back mutations. Amino acid residues that are candidates for back mutations are usually those located on the surface of the antibody molecule, while residues that are buried or have a low degree of surface exposure are generally not altered.

[0067] An alternative humanization technique for CDR grafting and back mutations is surface reshaping, in which non-surface-exposed residues of non-human origin are retained while surface residues are changed to human residues.

[0068] In some cases, it may be desirable to alter one or more CDR amino acid residues to improve the binding affinity with the target epitope. This is referred to as "affinity maturation". Various affinity maturation methods are known in the art, for example, the in vitro scanning saturation mutagenesis method described by Burks et al., Proc Natl Acad Sci USA, 94:412–417 (1997), and the stepwise in vitro affinity maturation method of Wu et al., Proc Natl Acad Sci USA 95:6037–6042 (1998).

[0069] The term "human antibody" refers to an antibody in which the variable domain and constant region sequences are derived from human sequences. The term encompasses antibodies having sequences derived from human genes, but those sequences have been modified, for example, to reduce immunogenicity, increase affinity, and increase stability. The term encompasses antibodies recombinantly produced in non-human cells, which may confer atypical glycosylation to human cells. The term also encompasses antibodies produced in transgenic non-human organisms having human antibody genes.

[0070] The "antigen-binding portion" or "antigen-binding fragment" of an antibody refers to the portion or fragment of the antibody that retains the ability to specifically bind to an antigen. In some embodiments, the antigen-binding fragment of the present disclosure is a Fab, Fab’, F(ab’)2, Fv or scFv fragment. In certain embodiments, the antigen-binding fragment of the present disclosure is an F(ab')2 fragment, which is a bivalent fragment comprising two Fab fragments linked in the hinge region by a disulfide bridge (Fab is composed of VL 、V H 、C L and C H1 domain - containing monovalent antibody fragments). In some embodiments, the antigen - binding fragments of the present disclosure may also contain C H2 or C H3 domains. Antibody fragments can be prepared using conventional techniques such as papain or pepsin digestion of intact antibodies. In addition, antibodies, antibody fragments, and immunoadhesion molecules can be obtained using, for example, standard recombinant DNA techniques as described herein.

[0071] The antibodies and antigen - binding fragments described herein can be isolated. The terms "isolated protein", "isolated polypeptide", or "isolated antibody" refer to such a protein, polypeptide, or antibody that, based on its origin or source of derivation, (1) is not associated with the natural associated components that accompany it in its natural state; (2) is substantially free of other proteins from the same species; (3) is expressed by cells from a different species; or (4) does not exist in nature. Thus, a polypeptide synthesized chemically or in a cell system different from the cell of its natural origin will be "isolated" from its natural associated components. By using protein purification techniques well - known in the art for isolation, a protein can also be made substantially free of natural associated components.

[0072] The class (isotype) and subclass of the antibodies described herein can be determined by any method known in the art. Generally, antibodies specific for a particular class and subclass of antibodies can be used to determine the class and subclass of an antibody. Such antibodies are commercially available. The class and subclass can be determined by ELISA, Western blotting, and other techniques. Alternatively, the class and subclass of an antibody can be determined by sequencing all or part of the constant region of the heavy and / or light chains of the antibody, comparing its amino acid sequence with the known amino acid sequences of the various classes and subclasses of immunoglobulins, and determining the class and subclass of the antibody. The preferred isotype of the present disclosure is the IgG isotype.

[0073] Agents that inhibit TGF - β

[0074] In some embodiments, the agent that specifically binds to TGF-β used in the combination therapies described herein is an anti-TGF-β antibody or an antigen-binding fragment thereof (e.g., Ab1, Fresolimumab, XOMA 089 / NIS793 (Gramont et al., Oncoimmunology 6(1):e1257453 (2017)), SRK-181 (Scholar Rock), ABBV-151 (AbbVie), lerdelimumab, or metelimumab) or a TGF-β trap molecule (e.g., M7824 (Knudson et al., Oncoimmunology 7(5):e1426519 (2018)) or AVID200 (Thwaites et al., Blood 130:2532 (2017))). In certain embodiments, the anti-TGF-β antibody is a human monoclonal antibody. In some embodiments, the anti-TGF-β antibody is a pan-TGF-β specific monoclonal antibody that is less prone to form half antibodies compared to previously known antibodies such as Fresolimumab. In some embodiments, the anti-TGF-β antibody has superior pharmacokinetic profiles, such as higher exposure in vivo, compared to Fresolimumab.

[0075] In some embodiments, the anti-TGF-β antibody is the antibody described in PCT patent publication WO 2018 / 134681, PCT patent publication WO 2006 / 086469, PCT patent publication WO 2014 / 153435, U.S. Patent 8,569,462, or U.S. Patent 7,527,791, which are incorporated herein by reference in their entirety. In certain embodiments, the anti-TGF-β antibody is antibody Ab1 or a variant thereof, where the variant may contain, for example, certain minimal amino acid changes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid changes, which may be in the framework region, for example) relative to Ab1 without loss of the antigen-binding specificity of the antibody.

[0076] Antibody Ab1 has an estimated molecular weight of 144 kD when unglycosylated. The amino acid sequences of its heavy and light chains are SEQ ID NO:1 and 2, respectively. These two sequences are shown below. The variable domains are italicized. The CDRs are boxed. The glycosylation site in the constant domain of the heavy chain is indicated in bold and lowercase (N297). Ab1 has a human IgG4 constant region, in which residue 228 (EU numbering) in the hinge region has been mutated from serine to proline. P228 is shown in boxed bold in the sequence of SEQ ID NO:1 below.

[0077]

[0078] In some embodiments, the anti-TGF-β antibodies or antigen-binding fragments described herein compete with Ab1 for binding to TGF-β or bind to the same epitope on TGF-β as Ab1.

[0079] In some embodiments, the anti-TGF-β antibodies described herein have a heavy chain comprising:

[0080] a) Heavy chain CDRs 1-3 (HCDR1-3) comprising the amino acid sequences of SEQ ID NOs: 5-7, respectively;

[0081] b) A heavy chain variable domain (V H ) that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO: 3;

[0082] c) A V H comprising the amino acid sequence of SEQ ID NO: 3; or

[0083] d) The amino acid sequence of SEQ ID NO: 1.

[0084] In some embodiments, the anti-TGF-β antibodies described herein have a light chain comprising:

[0085] a) Light chain CDRs 1-3 (LCDR1-3) comprising the amino acid sequences of SEQ ID NOs: 8-10, respectively;

[0086] b) A light chain variable domain (V L ) that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO: 4;

[0087] c) A V L comprising the amino acid sequence of SEQ ID NO: 4; or

[0088] d) The amino acid sequence of SEQ ID NO: 2.

[0089] In some embodiments, the anti-TGF-β antibodies described herein comprise any combination of any of the above heavy chains and any of the above light chains.

[0090] In some embodiments, the anti-TGF-β antibodies described herein comprise:

[0091] a) HCDR1-3 and LCDR1-3 comprising the amino acid sequences of SEQ ID NO:5-10, respectively;

[0092] b) a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO:3 H and a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO:4 L ;

[0093] c) a V comprising the amino acid sequence of SEQ ID NO:3 H and a V comprising the amino acid sequence of SEQ ID NO:4 L ; and

[0094] d) an HC comprising the amino acid sequence of SEQ ID NO:1 and an LC comprising the amino acid sequence of SEQ ID NO:2.

[0095] In some embodiments, the anti-TGF-β antibodies described herein (e.g., anti-TGF-β antibodies having the six CDRs of Ab1 or V H and V L ) have a human IgG4 constant region. In certain embodiments, residue 228 (EU numbering) in the hinge region of the IgG4 constant region has been mutated from serine to proline. The constant domains of the anti-TGF-β antibodies described herein can also be modified, for example, at Kabat residue L248 (e.g., by introducing the mutation L248E), to reduce any undesired effector functions of the molecule.

[0096] In some embodiments, the anti-TGF-β antibodies described herein have a human immunoglobulin κ light chain region.

[0097] In some embodiments, the anti-TGF-β antibodies or antigen-binding fragments described herein specifically bind to human TGF-β1, -β2, and -β3.

[0098] In some embodiments, the anti-TGF-β antibodies or antigen-binding fragments described herein bind at 1 x 10 -8 M or lower (e.g., 9 x 10 -9 M, 8 x 10 -9 M, 7 x 10 -9 M, 6 x 10 -9 M, 5 x 10 -9 M, 4 x 10 -9M, 3 x 10 -9 M, 2 x 10 -9 M or 1 x 10 -9 M) of K D in combination with TGF-β1, -β2 and / or -β3 (e.g., TGF-β1, -β2 and -β3). In certain embodiments, the anti-TGF-β antibody or antigen-binding fragment has a K of 3 x 10 -9 M or lower for binding D to TGF-β1, -β2 and -β3.

[0099] In some embodiments, the anti-TGF-β antibody or antigen-binding fragment described herein has one or more of the following properties:

[0100] a) inhibits TGF-β signal transduction;

[0101] b) neutralizes TGF-β when assayed in a mink lung epithelial cell assay;

[0102] c) has an EC50 of about 0.05 to 1 μg / ml as assayed in an A549 cell IL-11 induction assay;

[0103] d) inhibits the differentiation of CD4-positive T cells into inducible regulatory T cells (iTreg);

[0104] e) alleviates the immunosuppressive tumor microenvironment;

[0105] f) increases the MIP2 level in a patient (e.g., in the tumor tissue of a patient);

[0106] g) increases the KC / GRO level in a patient (e.g., in the tumor tissue of a patient);

[0107] h) promotes the activation or infiltration of tumor tissue by CD8-positive T cells (such as INF-γ-positive CD8-positive T cells);

[0108] i) increases the accumulation of natural killer (NK) cells in a patient (e.g., in the tumor tissue of a patient); and

[0109] j) restores the cytolytic activity of NK-92 cells after incubation with human recombinant TGF-β.

[0110] In some embodiments, the anti-TGF-β antibody or antigen-binding fragment described herein has 1, 2, 3, 4, 5, 6, 7, 8, 9 or all of the properties.

[0111] In some embodiments, compared to isatuximab, the anti-TGF-β antibodies or antigen-binding fragments described herein have an increased half-life, increased exposure, or both. For example, the increase is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% or more. The exposure of a drug (such as the antibodies or fragments described herein) is a function of the concentration of the drug in the body over time. The concentration of the drug in the body is typically indicated by the drug level in blood, plasma, or serum. The half-life and exposure (biological exposure) of the drug can be measured by well-known methods (e.g., as described in PCT patent publication WO 2018 / 134681).

[0112] An agent that inhibits CD38

[0113] In some embodiments, the agent that specifically binds to CD38 and is used in the combination therapies described herein is an anti-CD38 antibody or an antigen-binding fragment thereof. In certain embodiments, the anti-CD38 antibody is a humanized monoclonal antibody. In some embodiments, the anti-CD38 antibody is the antibody described in U.S. Patent 8,153,765, which is incorporated herein by reference in its entirety. In some embodiments, the anti-CD38 antibody is produced by a hybridoma cell line deposited at the American Type Culture Collection under accession number PTA-7670. In some embodiments, the anti-CD38 antibody is Ab2, Ab3, Ab4, daratumumab, MOR202 (Raab et al., Blood 128:1152 (2016)), TAK-079 (Roepcke et al., Pharmacol Res Perspect 6(3):e00402 (2018)), TAK-573 (Takeda), TAK-169 (Takeda), (Genmab / Janssen), anti-CD38 SIFbody (Momenta), or TSK011010 (CASI).

[0114] In certain embodiments, the anti-CD38 antibody is antibody Ab2 or a variant thereof, wherein the variant may contain, for example, certain minimal amino acid changes relative to Ab2 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid changes, which may be, for example, in the framework region) without loss of the antigen-binding specificity of the antibody.

[0115] The amino acid sequences of the heavy and light chains of Ab2 are SEQ ID NO:11 and 12, respectively. These two sequences are shown below. The variable domains are in italics. The CDRs are boxed. Ab2 has a human IgG1 constant region.

[0116]

[0117]

[0118] In some embodiments, the anti-CD38 antibody or antigen-binding fragment competes with Ab2 for binding to CD38 or binds to the same epitope on CD38 as Ab2.

[0119] In some embodiments, the anti-CD38 antibody has a heavy chain that comprises:

[0120] a) heavy chain CDR1-3 (HCDR1-3) containing the amino acid sequences of SEQ ID NO:15-17, respectively;

[0121] b) a heavy chain variable domain (V H ) that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO:13;

[0122] c) a V H containing the amino acid sequence of SEQ ID NO:13; or

[0123] d) the amino acid sequence of SEQ ID NO:11.

[0124] In some embodiments, the anti-CD38 antibody has a light chain that comprises:

[0125] a) light chain CDR1-3 (LCDR1-3) containing the amino acid sequences of SEQ ID NO:18-20, respectively;

[0126] b) a light chain variable domain (V L ) that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO:14;

[0127] c) a V L containing the amino acid sequence of SEQ ID NO:14; or

[0128] d) the amino acid sequence of SEQ ID NO:12.

[0129] In some embodiments, the anti-CD38 antibodies described herein comprise a combination of any of the above heavy chains and any of the above light chains.

[0130] In some embodiments, the anti-CD38 antibodies described herein comprise:

[0131] a) HCDR1-3 and LCDR1-3 containing the amino acid sequences of SEQ ID NOs: 15-20, respectively;

[0132] b) a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO: 13 H and a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO: 14 L ;

[0133] c) a V containing the amino acid sequence of SEQ ID NO: 13 H and a V containing the amino acid sequence of SEQ ID NO: 14 L ; and

[0134] d) an HC containing the amino acid sequence of SEQ ID NO: 11 and an LC containing the amino acid sequence of SEQ ID NO: 12.

[0135] In certain embodiments, the anti-CD38 antibody is antibody Ab3 or a variant thereof, wherein the variant may contain, for example, certain minimal amino acid changes relative to Ab3 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid changes, which may be, for example, in the framework regions) without loss of the antigen-binding specificity of the antibody.

[0136] The heavy and light chain amino acid sequences of Ab3 are SEQ ID NOs: 25 and 26, respectively. These two sequences are shown below. The variable domains are in italics. The CDRs are boxed.

[0137]

[0138] In some embodiments, the anti-CD38 antibody or antigen-binding fragment competes with Ab3 for binding to CD38 or binds to the same epitope on CD38 as Ab3.

[0139] In some embodiments, the anti-CD38 antibody has a heavy chain that comprises:

[0140] a) Heavy chain CDR1-3 (HCDR1-3) containing the amino acid sequences of SEQ ID NO: 29-31 respectively;

[0141] b) A heavy chain variable domain (V H ) that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 27;

[0142] c) V containing the amino acid sequence of SEQ ID NO: 27 H ; or

[0143] d) The amino acid sequence of SEQ ID NO: 25.

[0144] In some embodiments, the anti-CD38 antibody has a light chain, and the light chain comprises:

[0145] a) Light chain CDR1-3 (LCDR1-3) containing the amino acid sequences of SEQ ID NO: 32-34 respectively;

[0146] b) A light chain variable domain (V L ) that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 28;

[0147] c) V containing the amino acid sequence of SEQ ID NO: 28 L ; or

[0148] d) The amino acid sequence of SEQ ID NO: 26.

[0149] In some embodiments, the anti-CD38 antibody described herein comprises any combination of any of the above heavy chains and any of the above light chains.

[0150] In some embodiments, the anti-CD38 antibody described herein comprises:

[0151] a) HCDR1-3 and LCDR1-3 containing the amino acid sequences of SEQ ID NO: 29-34 respectively;

[0152] b) A V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO: 27 Hand a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:28 over the sequence L ;

[0153] c) a V comprising the amino acid sequence of SEQ ID NO:27 H and a V comprising the amino acid sequence of SEQ ID NO:28 L ; and

[0154] d) an HC comprising the amino acid sequence of SEQ ID NO:25 and an LC comprising the amino acid sequence of SEQ ID NO:26.

[0155] In certain embodiments, the anti-CD38 antibody is antibody Ab4 or a variant thereof, where the variant may contain, for example, certain minimal amino acid changes relative to Ab4 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid changes, which may be, for example, in the framework regions) without loss of the antigen-binding specificity of the antibody.

[0156] The heavy and light chain amino acid sequences of Ab4 are SEQ ID NO:35 and 36, respectively. These two sequences are shown below. The variable domains are in italics. The CDRs are boxed.

[0157]

[0158] In some embodiments, the anti-CD38 antibody or antigen-binding fragment competes with Ab4 for binding to CD38 or binds to the same epitope on CD38 as Ab4.

[0159] In some embodiments, the anti-CD38 antibody has a heavy chain that comprises:

[0160] a) heavy chain CDR1-3 (HCDR1-3) comprising the amino acid sequences of SEQ ID NOs: 39-41, respectively;

[0161] b) a heavy chain variable domain (V H ) that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of SEQ ID NO:37 over the sequence

[0162] c) a V comprising the amino acid sequence of SEQ ID NO:37 H ; or

[0163] d) The amino acid sequence of SEQ ID NO:35.

[0164] In some embodiments, the anti-CD38 antibody has a light chain, which comprises:

[0165] a) Light chain CDR1-3 (LCDR1-3) containing the amino acid sequences of SEQ ID NOs: 42-44, respectively;

[0166] b) A light chain variable domain (V L ) that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO:38;

[0167] c) V L containing the amino acid sequence of SEQ ID NO:38; or

[0168] d) The amino acid sequence of SEQ ID NO:36.

[0169] In some embodiments, the anti-CD38 antibody described herein comprises any combination of any of the above heavy chains and any of the above light chains.

[0170] In some embodiments, the anti-CD38 antibody described herein comprises:

[0171] a) HCDR1-3 and LCDR1-3 containing the amino acid sequences of SEQ ID NOs: 39-44, respectively;

[0172] b) A V H that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO:37 and a V L that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical in sequence to the amino acid sequence of SEQ ID NO:38;

[0173] c) V H containing the amino acid sequence of SEQ ID NO:37 and V L containing the amino acid sequence of SEQ ID NO:38; and

[0174] d) HC containing the amino acid sequence of SEQ ID NO:35 and LC containing the amino acid sequence of SEQ ID NO:36.

[0175] In some embodiments, the anti-CD38 antibodies described herein have a human IgG1 constant region.

[0176] In some embodiments, the anti-CD38 antibodies described herein have a human immunoglobulin kappa light chain region.

[0177] In some embodiments, the anti-CD38 antibody or antigen-binding fragment binds to CD38 with a K -8 of 1x10 -9 M or lower (e.g., 9x10 -9 M, 8x10 -9 M, 7x10 -9 M, 6x10 -9 M, 5x10 -9 M, 4x10 -9 M, 3x10 -9 M, or 1x10 -9 M). In certain embodiments, the anti-CD38 antibody or antigen-binding fragment binds to CD38 with a K D of 3x10 -9 M or lower. D

[0178] In some embodiments, the anti-CD38 antibody or antigen-binding fragment kills CD38-positive cells by apoptosis, ADCC, and CDC; in certain embodiments, killing of the CD38-positive cells by apoptosis can occur in the absence of stromal cells or stroma-derived cytokines. In some embodiments, the CD38-positive cells are malignant cells. In some embodiments, the CD38-positive cells are B cells. In certain embodiments, the CD38-positive cells are tumor cells derived from hematological malignancies. In more preferred embodiments, the CD38-positive cells are lymphoma cells, leukemia cells, or multiple myeloma cells. In another preferred embodiment, the CD38-positive cells are NHL, BL, MM, B-CLL, ALL, TCL, AML, HCL, HL, or CML cells.

[0179] In some embodiments, the anti-CD38 antibody or antigen-binding fragment is capable of killing at least 10%, 15%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, or 40% (e.g., at least 24%) of Daudi lymphoma cells in vitro in the absence of stromal cells or stroma-derived cytokines.

[0180] ​In some embodiments, the anti-CD38 antibodies or antigen-binding fragments described herein are capable of killing at least 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% (e.g., at least 7%) of Ramos lymphoma cells in vitro in the absence of stromal cells or stroma-derived cytokines.

[0181] In some embodiments, the anti-CD38 antibodies or antigen-binding fragments described herein are capable of killing at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% (e.g., at least 11%) of MOLP-8 multiple myeloma cells in vitro in the absence of stromal cells or stroma-derived cytokines.

[0182] In some embodiments, the anti-CD38 antibodies or antigen-binding fragments described herein are capable of killing at least 25%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 45%, or 50% (e.g., at least 36%) of SU-DHL-8 lymphoma cells in vitro in the absence of stromal cells or stroma-derived cytokines.

[0183] In some embodiments, the anti-CD38 antibodies or antigen-binding fragments described herein are capable of killing at least 20%, 25%, 26%, 27%, 28%, 29%, 30%, or 35% (e.g., at least 27%) of NU-DUL-1 lymphoma cells in vitro in the absence of stromal cells or stroma-derived cytokines.

[0184] In some embodiments, the anti-CD38 antibodies or antigen-binding fragments described herein are capable of killing at least 50%, 55%, 60%, 61%, 62%, 63%, 64%, 65%, 70%, or 75% (e.g., at least 62%) of DND-41 leukemia cells in vitro in the absence of stromal cells or stroma-derived cytokines.

[0185] In some embodiments, the anti-CD38 antibodies or antigen-binding fragments described herein are capable of killing at least 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, or 13% (e.g., at least 9%) of JVM-13 leukemia cells in vitro in the absence of stromal cells or stroma-derived cytokines.

[0186] In some embodiments, the anti-CD38 antibodies or antigen-binding fragments described herein are capable of killing at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, or 8% (e.g., at least 4%) of HC-1 leukemia cells in vitro in the absence of stromal cells or stroma-derived cytokines.

[0187] In some embodiments, the agent that specifically binds to CD38 is a conjugate comprising an anti-CD38 antibody or antigen-binding fragment described herein linked to a cytotoxic agent. The cytotoxic agent can be selected from, for example, maytansine alkaloids, small molecules, tomaymycin derivatives, leptomycin derivatives, prodrugs, taxanes, CC-1065 and CC-1065 analogs, or any cytotoxic agent described in U.S. Patent 8,153,765, which is incorporated herein by reference in its entirety.

[0188] The class of the anti-TGF-β or anti-CD38 antibodies described herein can be altered or converted into another class or subclass. In one aspect, nucleic acid molecules encoding V L or V H are isolated using methods well known in the art such that they do not contain nucleic acid sequences encoding CL or CH. The nucleic acid molecules encoding V L or V H are then operably linked to nucleic acid sequences encoding CL or CH from immunoglobulin molecules of different classes. As described above, this can be achieved using a vector or nucleic acid molecule comprising the CL or CH chain. For example, an antibody initially of IgM can be class-converted to IgG. Additionally, the class switch can be used to convert one IgG subclass to another, e.g., from IgG1 to IgG2. The κ light chain constant region can be altered to the λ light chain constant region, for example. Preferred methods for generating antibodies as described herein with a desired Ig isotype include the steps of isolating a nucleic acid molecule encoding the heavy chain of the antibody and a nucleic acid molecule encoding the light chain of the antibody, obtaining the variable domain of the heavy chain, linking the variable domain of the heavy chain to the constant region of the heavy chain of the desired isotype, expressing the light chain and the linked heavy chain in a cell, and collecting the antibody having the desired isotype.

[0189] The antibodies described herein can be IgG, IgM, IgE, IgA, or IgD molecules, but typically belong to the IgG isotype, e.g., to the IgG subclasses IgG1, IgG 2a or IgG 2b , IgG3, or IgG4.

[0190] In one embodiment, the antibody can comprise at least one mutation in the Fc region. Many different Fc mutations are known, and these mutations provide altered effector functions. For example, in many cases, it will be desirable to reduce or eliminate effector function, such as in cases where ligand / receptor interactions are not desired or in the case of antibody-drug conjugates.

[0191] In some embodiments, the antibodies described herein (such as anti-TGF-β and anti-CD38 antibodies) do not have a C-terminal lysine in the heavy chain. The C-terminal lysine can be removed during the manufacturing process or by recombinant techniques (i.e., the coding sequence of the heavy chain does not contain a codon for the C-terminal lysine). Accordingly, antibodies comprising the heavy chain amino acid sequence SEQ ID NO:1 or 11 that do not have a C-terminal lysine are also contemplated within the present disclosure.

[0192] Combination therapy

[0193] The present disclosure provides a combination therapy comprising an agent that specifically binds to human TGF-β and an agent that specifically binds to human CD38. In some embodiments, the agent that specifically binds to TGF-β can be any anti-TGF-β antibody or antigen-binding fragment thereof described herein. In some embodiments, the agent that specifically binds to CD38 can be any anti-CD38 antibody or antigen-binding fragment thereof described herein. The present disclosure also contemplates combination therapies having one or more other agents that inhibit TGF-β (e.g., galunisertib, LY3200882, PF-06952229 (Pfizer), GFH-018 (GenFleet), and / or vactosertib) and / or one or more other agents that inhibit CD38. The combination therapies described herein can take the form of a method of treatment using the agents or pharmaceutical compositions comprising the agents.

[0194] The present disclosure also contemplates a combination therapy comprising an agent that specifically binds to human CD38 (e.g., an anti-CD38 antibody or antigen-binding fragment thereof, such as one described herein) and an agent that specifically binds to LAP (e.g., an anti-LAP antibody).

[0195] In some embodiments, the combination therapies of the present disclosure use an anti-TGF-β antibody Ab1 and an anti-CD38 antibody Ab2. In some embodiments, the combination therapies of the present disclosure use an antibody or antigen-binding fragment thereof that competes with Ab1 for binding to TGF-β or binds to the same epitope of TGF-β as Ab1 and an antibody that competes with Ab2 for binding to CD38 or binds to the same epitope of CD38 as Ab2. In certain embodiments, the Ab1 and Ab2 agents are used in separate compositions (e.g., administered sequentially). In certain embodiments, the Ab1 and Ab2 agents are used in a single composition.

[0196] In certain embodiments, the combination therapies of the present disclosure use:

[0197] - an anti-TGF-β antibody or antigen-binding fragment thereof comprising HCDR1-3 and LCDR1-3 having the amino acid sequences of SEQ ID NOs: 5-10, respectively; and an anti-CD38 antibody comprising HCDR1-3 and LCDR1-3 having the amino acid sequences of SEQ ID NOs: 15-20, respectively;

[0198] - an anti-TGF-β antibody or antigen-binding fragment thereof comprising: a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in sequence to the amino acid sequence of SEQ ID NO:3 H and a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in sequence to the amino acid sequence of SEQ ID NO:4; and an anti-CD38 antibody comprising: a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in sequence to the amino acid sequence of SEQ ID NO:13 L and a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in sequence to the amino acid sequence of SEQ ID NO:14; H and a V that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical in sequence to the amino acid sequence of SEQ ID NO:14; L ;

[0199] - an anti-TGF-β antibody or antigen-binding fragment thereof comprising V having the amino acid sequences of SEQ ID NOs: 3 and 4, respectively H and V L and an anti-CD38 antibody comprising V having the amino acid sequences of SEQ ID NOs: 13 and 14, respectively H and VL an anti-CD38 antibody; or

[0200] - an anti-TGF-β antibody comprising an HC and an LC having the amino acid sequences of SEQ ID NOs: 1 and 2, respectively; and an anti-CD38 antibody comprising an HC and an LC having the amino acid sequences of SEQ ID NOs: 11 and 12, respectively.

[0201] In some embodiments, the anti-TGF-β antibody and the anti-CD38 antibody used in the combination therapy have subtypes IgG4 and IgG1, respectively. In certain embodiments, the anti-TGF-β antibody has a human IgG4 constant region, wherein residue 228 (EU numbering) in the hinge region has been mutated from serine to proline.

[0202] In some embodiments, the anti-TGF-β antibody or an antigen-binding fragment thereof described herein alleviates the immunosuppressive effect of TGF-β on NK cell-mediated ADCC by the anti-CD38 antibody described herein, such that the combination therapy of the present disclosure is more effective than treatment with the anti-CD38 antibody alone.

[0203] In some embodiments, a combination therapy comprising an agent that specifically binds to human TGF-β and an agent that specifically binds to human CD38 (e.g., an anti-TGF-β antibody and an anti-CD38 antibody, such as Ab1 and Ab2, respectively) may further comprise an additional agent or therapy. In certain embodiments, the additional agent or therapy may be, for example, lenalidomide, pomalidomide, bortezomib, methylprednisolone, dexamethasone, prednisone, melphalan, bortezomib, ixazomib, carfilzomib, thalidomide, cyclophosphamide, pembrolizumab, a pan-histone deacetylase inhibitor (e.g., Panobinostat), retinoic acid (e.g., all-trans retinoic acid), radioimmunotherapy, chemotherapy, etc. In certain embodiments, the additional agent or therapy is a combination of lenalidomide and dexamethasone. In certain embodiments, the additional agent or therapy is a combination of bortezomib, melphalan, and prednisone. In certain embodiments, the additional agent or therapy is a combination of bortezomib and dexamethasone. In certain embodiments, the additional agent or therapy is a combination of pomalidomide and dexamethasone. In certain embodiments, the additional agent or therapy is dexamethasone. In some embodiments, the additional agent or therapy is treatment for the disorder targeted by the combination therapy of the present disclosure. For example, when the disorder is myeloma (e.g., multiple myeloma), the treatment may be, for example, elotuzumab.

[0204] In some embodiments, the agents in the combination therapies of the present disclosure are administered in more than one composition. In certain embodiments, each agent is provided in a separate composition. In the case where there is more than one composition, the compositions can be administered simultaneously, sequentially, or separately. In other embodiments, the agents are administered in a single composition. For example, a combination therapy comprising an anti-TGF-β antibody and an anti-CD38 antibody can involve administering a single composition comprising both antibodies, or separate compositions for each antibody (where the separate compositions can be administered sequentially or in parallel).

[0205] Therapeutic use of the combination therapies of the present disclosure

[0206] In one aspect, the combination therapies of the present disclosure are used to treat a disorder that is dependent on CD38 expression. In some embodiments, the combination therapies of the present disclosure are used to treat a hyperproliferative disorder, an inflammatory disease, an autoimmune disease, or a fibrotic disorder. In certain embodiments, the combination therapies of the present disclosure are used to treat cancer.

[0207] In some embodiments, the combination therapies of the present disclosure target CD38-positive cells (e.g., CD38-positive cancer cells, such as malignant B cells). Cells can be identified as CD38-positive by any suitable method for determining gene or protein expression, such as by histology, flow cytometry, RT-PCR, or RNA-Seq. The cancer cells for determination can be obtained by tumor biopsy or by collecting circulating tumor cells. Without wishing to be bound by theory, it is contemplated that an agent that specifically binds to CD38 will bind to CD38-positive cells and mediate ADCC / CDC on said cells, and an agent that specifically binds to TGF-β will reduce the immunosuppressive effect of TGF-β, thereby enhancing the efficacy of the cancer therapy.

[0208] In some embodiments, the combination therapies of the present disclosure are used to treat myeloma, such as multiple myeloma (e.g., relapsed and / or refractory multiple myeloma, newly diagnosed multiple myeloma (optionally not suitable for transplantation), smoldering multiple myeloma, light chain myeloma, non-secretory myeloma, immunoglobulin D myeloma, or immunoglobulin E myeloma). In certain embodiments, compared to a therapy using only an agent that specifically binds to CD38 (e.g., an anti-CD38 antibody), the combination therapies of the present disclosure result in less bone destruction in myeloma bone lesions. In specific embodiments, the combination therapies of the present disclosure can result in improved healing of myeloma bone lesions. Without wishing to be bound by theory, it is contemplated that an agent that specifically binds to TGF-β (e.g., an anti-TGF-β antibody) will inhibit the inhibitory activity of TGF-β on osteoblast differentiation and matrix mineralization, thereby enhancing bone formation and resulting in bone remodeling and bone healing. Additionally, because mature osteoblasts enhance apoptosis and cell cycle arrest of multiple myeloma cells, an agent that specifically binds to TGF-β can also inhibit multiple myeloma cell growth.

[0209] In some embodiments, the combination therapies of the present disclosure are used to treat, for example, amyloidosis (such as relapsed or refractory primary amyloidosis or light chain amyloidosis), myelodysplastic syndromes (MDS), monoclonal gammopathy, solitary plasmacytoma, extramedullary plasmacytoma, or Waldenström macroglobulinemia).

[0210] In some embodiments, the combination therapies of the present disclosure are used to treat hematological malignancies, such as leukemia or lymphoma. For example, the malignancies can be chronic lymphocytic leukemia, B and T acute lymphocytic leukemia, acute lymphocytic leukemia (e.g., B cell acute lymphocytic leukemia or B cell or T cell precursor acute lymphocytic leukemia), chronic lymphocytic acute myeloid leukemia, chronic myeloid leukemia, acute myeloid leukemia, chronic myeloid leukemia, promyelocytic leukemia, and hairy cell leukemia), non-Hodgkin lymphoma, Hodgkin lymphoma, Burkitt lymphoma, B cell lymphoma (such as diffuse large B cell lymphoma or germinal center B cell lymphoma), T cell lymphoma (such as peripheral T cell lymphoma), natural killer / T cell lymphoma (e.g., nasal type), lymphoblastic lymphoma, mantle cell lymphoma, and follicular lymphoma. In certain embodiments, the combination therapies of the present disclosure are used to treat lymphoblastic leukemia.

[0211] Other cancers that can be treated by the combination therapies of the present disclosure can be solid tumors and can include, but are not limited to, skin cancer (e.g., melanoma (including unresectable or metastatic melanoma), cutaneous squamous cell carcinoma, xeroderma pigmentosum, and keratoacanthoma), thyroid cancer, lung cancer (e.g., non-small cell lung cancer), esophageal cancer, gastric cancer, colon cancer, colorectal cancer, pancreatic cancer, liver cancer (e.g., hepatocellular carcinoma), primary peritoneal cancer, bladder cancer, kidney cancer or renal carcinoma (e.g., renal cell carcinoma), urothelial cancer, breast cancer (e.g., Her2-positive breast cancer or triple-negative breast cancer), ovarian cancer, fallopian tube cancer, cervical cancer, uterine cancer, prostate cancer, testicular cancer, head and neck cancer (e.g., head and neck squamous cell carcinoma), brain cancer, neuroblastoma, glioblastoma, glioma, astrocytoma, schwannoma, mesothelioma, fibrosarcoma, rhabdomyosarcoma, osteosarcoma, Kaposi’s sarcoma, seminoma, and teratocarinoma. In some embodiments, the combination therapies of the present disclosure are used to treat non-small cell lung cancer, prostate cancer (e.g., prostate adenocarcinoma), glioblastoma, hepatocellular carcinoma, ovarian cancer, head and neck cancer, urothelial cancer, or colorectal cancer.

[0212] The combination therapies of the present disclosure can also be used to inhibit cyclosporine-mediated malignancy or cancer progression (e.g., metastasis).

[0213] In some embodiments, the combination therapies of the present disclosure can be used to treat cancer at an early, intermediate, advanced, or metastatic stage. In some embodiments, the combination therapy is used to treat patients who have received at least one prior therapy (e.g., patients with multiple myeloma). In some embodiments, the combination therapies can be used to treat recurrent or refractory cancer.

[0214] In some embodiments, the combination therapies of the present disclosure are used to treat any of the conditions described herein (e.g., the cancers described herein) in patients who have progressed during or after standard therapy for the condition or for whom there is no effective standard therapy for the condition.

[0215] In some embodiments, the combination therapies of the present disclosure are used to treat any of the conditions described herein (e.g., the cancers described herein) in patients who are resistant to one or more standard therapies for the condition. In certain embodiments, the patient may have a resistance to Ab2, Ab3, Ab4, carfilzomib, LY3200882, XOMA 089, daratumumab, MOR202, TAK-079, TAK-573, TAK-169, or any combination thereof is a refractory disorder (e.g., cancer such as multiple myeloma). The disorder may be refractory to treatment with an agent that specifically binds to CD38. In certain embodiments, the patient may have a disorder (e.g., cancer such as multiple myeloma) that is refractory to treatment with Ab2 or with daratumumab, or refractory to both treatments.

[0216] In some embodiments, the combination therapies of the present disclosure are used to treat the cancers described herein, wherein the cancers exhibit high levels of TGF-β expression. In certain embodiments, the cancers exhibit high levels of TGF-β expression and are resistant to treatment with an agent that specifically binds to CD38 (e.g., Ab2, Ab3, Ab4, daratumumab, MOR202, TAK-079, TAK-573, TAK-169, or any combination thereof). In certain embodiments, the cancer may be refractory to treatment with Ab2 or with daratumumab, or refractory to both treatments.

[0217] In some embodiments, the combination therapies of the present disclosure are used to treat patients with newly diagnosed multiple myeloma who are unable to receive a type of stem cell transplantation (autologous stem cell transplantation) using their own stem cells. Additionally or alternatively, the patient has received at least one prior drug for the treatment of multiple myeloma. In certain embodiments, the combination therapy further comprises:

[0218] a) lenalidomide and / or dexamethasone,

[0219] b) bortezomib, lenalidomide and / or dexamethasone,

[0220] c) bortezomib, melphalan and / or prednisone, or

[0221] d) bortezomib and / or dexamethasone.

[0222] In some embodiments, the combination therapies of the present disclosure are used to treat patients who have received at least two prior drugs for the treatment of multiple myeloma. In certain embodiments, the two prior drugs include lenalidomide and / or a proteasome inhibitor.

[0223] In some embodiments, the combination therapies of the present disclosure are used to treat patients who have received at least three prior drugs for the treatment of multiple myeloma. In certain embodiments, the three prior drugs include a proteasome inhibitor and / or an immunomodulatory agent.

[0224] In some embodiments, the combination therapies of the present disclosure are used to treat patients who are unresponsive to proteasome inhibitors and / or immunomodulators.

[0225] In some embodiments, the combination therapies of the present disclosure are used to treat patients with relapsed / refractory multiple myeloma in combination with pomalidomide and / or dexamethasone (e.g., low-dose dexamethasone). In certain embodiments, the patient has received at least two prior drugs for the treatment of multiple myeloma. In specific embodiments, the two prior drugs include lenalidomide and / or a proteasome inhibitor.

[0226] "Treat", "treating", and "treatment" refer to a method of alleviating or eliminating at least one of a biological disorder and / or its accompanying symptoms. As used herein, "alleviating" a disease, disorder, or condition means reducing the severity and / or frequency of the symptoms of the disease, disorder, or condition. In addition, references to "treatment" herein include references to curative, palliative, and prophylactic treatment. It should of course be understood that in the context of cancer therapy, "treatment" includes any medical intervention that results in a slowdown in cancer growth, a delay in cancer progression or recurrence, or a reduction in cancer metastasis, as well as a partial remission of cancer to extend the patient's life expectancy.

[0227] As used herein, the terms "co-administration" and "in combination with" refer to, but are not limited to, (i) administering such agents simultaneously to a patient in need of treatment when the therapeutic agents are formulated together as a single dosage form, (ii) administering such agents substantially simultaneously to a patient in need of treatment when such agents are formulated separately into separate dosage forms, and (iii) administering such agents sequentially to a patient in need of treatment when such agents are formulated separately into separate dosage forms to be taken by the patient at different times.

[0228] The ratio between an agent that specifically binds to TGF-β (e.g., an anti-TGF-β antibody) and an agent that specifically binds to CD38 (e.g., an anti-CD38 antibody) can be a ratio such that the agents are administered in equal amounts (i.e., a 1:1 ratio), but this is not necessarily the case. Depending on the characteristics of the individual agents, it may be desirable to use non-equal amounts of the agents.

[0229] It should be understood that the combination therapies of the present disclosure can be used in the treatment methods as described herein, can be used in the treatment as described herein, and / or can be used in the manufacture of a medicament for the treatment as described herein.

[0230] Dosing regimen

[0231] The combination therapies of the present disclosure will be administered in an effective amount for treating the disorder under discussion, i.e., the dosage and time period required to achieve the desired result. The therapeutically effective amount can vary depending on a variety of factors such as the particular disorder being treated, the age, sex, health status and weight of the patient, and whether the agent is administered as a single therapy or in combination with one or more additional anti-cancer therapies.

[0232] "Therapeutically effective amount" refers to the amount of a therapeutic agent that will, to some extent, alleviate one or more symptoms of the disorder being treated. This amount can be determined by healthcare professionals using well-established principles. A therapeutically effective amount of an anti-cancer therapeutic agent can, for example, result in tumor shrinkage, increased survival, elimination of cancer cells, reduced disease progression, reversal of metastasis, or other clinical endpoints desired by healthcare professionals.

[0233] In some embodiments, when treating with the combination therapies of the present disclosure, the cardiac and pulmonary side effects of the patient are monitored.

[0234] Host cells and methods for producing antibodies and antibody compositions

[0235] One aspect of the present disclosure relates to methods for producing antibodies for use in the combination therapies of the present invention. One embodiment relates to a method for producing an antibody as described herein, the method comprising providing a recombinant host cell capable of expressing the antibody, culturing the host cell under conditions suitable for antibody expression, and isolating the resulting antibody. The antibody produced by such expression in such recombinant host cells is referred to herein as a "recombinant antibody". Also described are progeny cells of such host cells, and antibodies produced by such progeny cells.

[0236] As used herein, the term "recombinant host cell" (or simply "host cell") means a cell into which a recombinant expression vector has been introduced. The host cell can comprise, for example, one or more of the vectors described herein. The host cell can comprise, for example, a nucleotide sequence encoding the heavy chain of an anti-TGF-β and / or anti-CD38 antibody or an antigen-binding fragment thereof as described herein, or an antigen-binding fragment of the heavy chain, a nucleotide sequence encoding the light chain of an anti-TGF-β and / or anti-CD38 antibody or an antigen-binding fragment thereof as described herein, or an antigen-binding fragment of the light chain, or both. It should be understood that "recombinant host cell" and "host cell" refer not only to a particular subject cell, but also to progeny of such a cell. Because certain modifications may occur in progeny due to mutation or environmental influences, such progeny may actually be different from the parental cell, but are still included within the scope of the term "host cell" as used herein.

[0237] In some embodiments, the host cells of the present disclosure comprise:

[0238] - A nucleotide sequence encoding a heavy chain of an anti-TGF-β antibody or an antigen-binding fragment thereof as described herein, a nucleotide sequence encoding a light chain of an anti-TGF-β antibody or an antigen-binding fragment thereof as described herein, or both, and

[0239] - A nucleotide sequence encoding a heavy chain of an anti-CD38 antibody or an antigen-binding fragment thereof as described herein, a nucleotide sequence encoding a light chain of an anti-CD38 antibody or an antigen-binding fragment thereof as described herein, or both.

[0240] Nucleic acid molecules encoding the amino acid sequences of the heavy and / or light chains of anti-TGF-β and / or anti-CD38 antibodies or antigen-binding fragments thereof may be included in an expression vector. Expression vectors in which the nucleic acid sequence of interest is linked to the necessary expression control sequences, such as transcriptional and translational control sequences, include plasmids, retroviruses, adenoviruses, adeno-associated viruses (AAV), plant viruses (such as cauliflower mosaic virus), tobacco mosaic virus, cosmids, YACs, EBV-derived episomes, etc. The antibody light chain coding sequence and the antibody heavy chain coding sequence may be inserted into separate vectors and may be operably linked to the same or different expression control sequences (e.g., promoters). In one embodiment, both coding sequences are inserted into the same expression vector and may be operably linked to the same expression control sequence (e.g., a common promoter), separate identical expression control sequences (e.g., multiple promoters), or different expression control sequences (e.g., multiple promoters). The antibody coding sequences are inserted into the expression vector by standard methods (e.g., ligating the antibody gene fragment to complementary restriction sites on the vector or blunt-end ligation when there are no restriction sites).

[0241] Expression vectors encoding the antibodies and antigen-binding fragments described herein can be introduced into host cells for expression. In some embodiments, an expression vector encoding an anti-TGF-β antibody and an expression vector encoding an anti-CD38 antibody are introduced into separate host cells. In other embodiments, the expression vectors are introduced into the same host cell. The host cells are cultured under conditions suitable for antibody expression and then harvested and isolated. Host cells include mammalian, plant, bacterial, or yeast host cells. Mammalian cell lines that are available as hosts for expression are well known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC). These include, in particular, Chinese hamster ovary (CHO) cells, NS0 cells, SP2 cells, HEK-293T cells, 293Freestyle cells (Invitrogen), NIH-3T3 cells, HeLa cells, baby hamster kidney (BHK) cells, African green monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, and many other cell lines. The cell line can be selected based on the expression level. Other cell lines that can be used are insect cell lines, such as Sf9 or Sf21 cells.

[0242] In addition, a variety of known techniques can be used to enhance antibody expression. For example, the glutamine synthetase gene expression system (GS system) is a commonly used method for enhancing expression under certain conditions.

[0243] The tissue culture medium for the host cells can contain or not contain animal-derived components (ADCs), such as bovine serum albumin. In some embodiments, a medium without ADCs is preferred for human safety. Tissue culture can be carried out using a fed-batch method, a continuous perfusion method, or any other method suitable for the host cells and the desired yield.

[0244] In some embodiments, the present disclosure relates to a method for producing an antibody composition comprising an anti-TGF-β antibody or an antigen-binding fragment thereof and an anti-CD38 antibody or an antigen-binding fragment thereof, the method comprising:

[0245] - providing a first host cell and a second host cell, wherein the first host cell is capable of expressing an anti-TGF-β antibody or an antigen-binding fragment thereof as described herein, and the second host cell is capable of expressing an anti-CD38 antibody or an antigen-binding fragment thereof as described herein,

[0246] - culturing the first host cell and the second host cell under conditions suitable for the expression of the anti-TGF-β antibody or an antigen-binding fragment thereof and the anti-CD38 antibody or an antigen-binding fragment thereof,

[0247] - isolating the resulting antibody or antigen-binding fragment, and

[0248] Optionally, combine the antibody or antigen-binding fragment to produce an antibody composition.

[0249] Pharmaceutical composition

[0250] Another aspect of the present disclosure is a pharmaceutical composition comprising an agent that specifically binds to human TGF-β (e.g., an anti-TGF-β antibody or an antigen-binding fragment thereof) and an agent that specifically binds to human CD38 (e.g., an anti-CD38 antibody or an antigen-binding fragment thereof) as active ingredients (e.g., as the sole active ingredient). The agent that specifically binds to TGF-β and the agent that specifically binds to CD38 can be co-formulated, e.g., mixed and provided in a single composition. The present disclosure also provides (1) a pharmaceutical composition comprising an agent that specifically binds to human TGF-β, and (2) a pharmaceutical composition comprising an agent that specifically binds to human CD38, wherein the pharmaceutical compositions are used in the same combination therapy.

[0251] In some embodiments, when administered in the combination therapy of the present disclosure, the pharmaceutical compositions described herein are intended for treating (e.g., ameliorating and / or preventing) a disorder, disease or condition, the treatment of which improves or slows the progression of the disorder, disease or condition by modulating the activity or expression of TGF-β and CD38. In certain embodiments, the pharmaceutical compositions are intended for treating (e.g., ameliorating and / or preventing) cancer. In a particular embodiment, the cancer is multiple myeloma.

[0252] In some embodiments, the pharmaceutical compositions of the present disclosure comprise the anti-TGF-β antibody described herein, which has less than 1% half-antibody. The formation of half-antibody can be determined by purity analysis of monoclonal antibody preparations, which is carried out by, for example, SDS-capillary electrophoresis or non-reducing SDS-PAGE analysis under non-reducing conditions, followed by densitometry or RP-HPLC (Angal et al., Mol Immunol 30(1):105-8 (1993); Bloom et al., Protein Science 6:407-415 (1997); Schuurman et al., 38(1):1-8 (2001); and Solanos et al., Anal Chem 78:6583-94 (2006)).

[0253] Generally, the pharmaceutical compositions described herein are suitable for administration as a formulation in combination with one or more pharmaceutically acceptable excipients (e.g., as described below).

[0254] As used herein, the term "excipient" or "carrier" describes any component other than one or more compounds of the present disclosure. The choice of excipient will depend to a large extent on various factors, such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form. "Pharmaceutically acceptable excipients" include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc., that are physiologically compatible. Some examples of pharmaceutically acceptable excipients are water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, etc., and combinations thereof. In some cases, an isotonic agent, such as sugar, polyols (e.g., mannitol, sorbitol) or sodium chloride, will be included in the composition. Additional examples of pharmaceutically acceptable substances are wetting agents or minor auxiliary substances (such as wetting agents or emulsifying agents, preservatives or buffers) that increase the shelf life or effectiveness of the antibody. In some embodiments, the pharmaceutical compositions of the present disclosure may comprise hyaluronidase (e.g., recombinant human hyaluronidase). In certain embodiments, pharmaceutical compositions comprising hyaluronidase may be used for subcutaneous administration.

[0255] The pharmaceutical compositions of the present disclosure may be prepared, packaged, or provided as a single unit dose or as multiple single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition that contains a predetermined amount of one or more active ingredients. The amount of each active ingredient is usually equal to the dose of the active ingredient to be administered to a subject or a convenient fraction of such a dose, such as one-half or one-third of such a dose.

[0256] The pharmaceutical compositions of the present disclosure are generally suitable for parenteral administration. As used herein, "parenteral administration" of a pharmaceutical composition includes any route of administration characterized by physically making an opening in the tissue of a subject and administering the pharmaceutical composition through the opening in the tissue, thus generally resulting in direct administration into the blood, muscle, or internal organ. Thus, parenteral administration includes, but is not limited to, administering the composition by injection, applying the composition through a surgical incision, applying the composition through a non-surgical wound that penetrates the tissue, etc. In particular, parenteral administration is contemplated to include, but is not limited to, subcutaneous, intraperitoneal, intramuscular, intrasternal, intravenous, intraarterial, intrathecal, intraventricular, intraurethral, intracranial, intratumoral, and intrasynovial injection or infusion; and kidney dialysis infusion techniques. Local perfusion is also contemplated. Preferred embodiments may include intravenous and subcutaneous routes.

[0257] Formulations of pharmaceutical compositions suitable for parenteral administration generally comprise one or more active ingredients in combination with a pharmaceutically acceptable carrier such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or presented in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or presented in unit dosage form, such as in an ampoule or in a multi-dose container containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and the like. Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending agents, stabilizers, or dispersing agents. In one embodiment of a formulation for parenteral administration, one or more active ingredients are provided in a dry (i.e., powder or granule) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) and then the reconstituted composition is administered parenterally. Parenteral formulations also include aqueous solutions that may contain excipients such as salts, carbohydrates, and buffers (e.g., having a pH of from 3 to 9), but for some applications they may be more suitably formulated as sterile non-aqueous solutions or in a dry (e.g., lyophilized) form to be combined with a suitable vehicle (such as sterile pyrogen-free water). Exemplary parenteral administration forms include solutions or suspensions in the form of sterile aqueous solutions (e.g., aqueous propylene glycol or glucose solutions). Such dosage forms may be buffered appropriately if desired. Other available parenteral administration formulations include those comprising one or more active ingredients in microcrystalline form or in liposomal formulations. Formulations for parenteral administration may be formulated for immediate release and / or modified release. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release, and programmed release.

[0258] Articles and kits

[0259] The present disclosure also provides an article comprising an agent that specifically binds to human TGF-β and an agent that specifically binds to human CD38. In some embodiments, the article of the present disclosure comprises an anti-TGF-β antibody or an antigen-binding fragment thereof as described herein, and an anti-CD38 antibody or an antigen-binding fragment thereof as described herein. In certain embodiments, the article comprises Ab1 and Ab2. The present disclosure further provides a method for manufacturing the article.

[0260] The present disclosure also provides a kit comprising an agent that specifically binds to human TGF-β and an agent that specifically binds to human CD38, and instructions for use in combination with the agents. In some embodiments, the kit of the present disclosure comprises an anti-TGF-β antibody or an antigen-binding fragment thereof as described herein, and an anti-CD38 antibody or an antigen-binding fragment thereof as described herein. In certain embodiments, the kit comprises Ab1 and Ab2.

[0261] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings commonly understood by one of ordinary skill in the art. Exemplary methods and materials are described below, but methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure. In case of conflict, the present specification, including definitions, will control.

[0262] Generally, the nomenclature and techniques used herein in connection with cell and tissue culture, molecular biology, immunology, microbiology, genetics, analytical chemistry, synthetic organic chemistry, medicine and pharmaceutical chemistry, and protein and nucleic acid chemistry and hybridization are those well known and commonly used in the art. Enzyme reactions and purification techniques are performed according to the manufacturer's instructions, as commonly practiced in the art or as described herein.

[0263] Furthermore, unless otherwise required by context, singular terms shall include the plural and plural terms shall include the singular. Throughout this specification and the embodiments, the words "have" and "comprise" or variations such as "has", "having", "comprises" or "comprising" shall be understood to imply the inclusion of the stated integer or group of integers, but not the exclusion of any other integer or group of integers.

[0264] All publications and other references mentioned herein are incorporated by reference in their entirety. Although many documents are cited herein, such citation does not mean that any of these documents constitutes a part of the common general knowledge in the art.

[0265] Examples

[0266] To better understand the present disclosure, the following examples are set forth. These examples are for illustrative purposes only and should not be construed in any way as limiting the scope of the present disclosure.

[0267] Example 1: Release of TGF-β by Multiple Myeloma and Lymphoma Cell Lines

[0268] JJN3, NCI-H929, RPMI8226, LP1, MOLP8, SUDHL-4, DAUDI, OCI-LY19, and SUDHL8 cells were cultured in RPMI 1640 (Invitrogen catalog number 22400-089) supplemented with 10% fetal bovine serum (Invitrogen catalog number 10082-147).

[0269] To quantify TGF-β released by MM and lymphoma cell lines, 1 x 106 Cells were plated in a volume of 100 μL of RPMI supplemented with 10% fetal bovine serum in a 96-well tissue culture-treated plate. The cells were incubated at 37 °C in a humidified incubator with 5% CO2 for 1 day, 2 days, or 3 days. After centrifugation at 250 g for 5 minutes, 40 μL of supernatant was collected from each well, and the total human TGF-β level was measured using the MSD 96-well multi-array human TGF-β1 assay kit (Meso Scale Discovery catalog number K151IUC-1) according to the manufacturer's instructions. Briefly, the samples were activated by acid treatment and then neutralized, after which they were incubated with the capture antibody. The antibody was then detected using an ELISA-based method, and the results were read on an MSD SECTOR imager.

[0270] Total TGF-β content was detected at levels exceeding 500 pg / mL in the supernatants of all MM and lymphoma cell lines, and accumulation over time was observed in most cell lines, particularly in JJN3, RPMI8226, and MOLP8 cells (Figure 1).

[0271] Example 2: TGF-β reduces the cytolytic activity of human NK cells and diminishes Ab2-mediated ADCC

[0272] Logarithmically growing NK92V cells (NK92-05 CD16 V / V; Conkwest) were suspended in growth medium (Middle MyeloCult H5100 (Stemcell Technologies catalog number 05150) supplemented with 100 units / mL of human interleukin-2 (R&D catalog number 202-IL-010 / CF)) and seeded into 10 cm culture dishes at a density of 0.8x10 5 cells / mL. The cells were treated with recombinant human TGF-β at a final concentration of 10 ng / mL at 37 °C for overnight, 24 hours, 48 hours, or 72 hours.

[0273] MOLP8 cells were cultured in RPMI1640 (Invitrogen catalog number 22400-089) supplemented with 20% fetal bovine serum (Invitrogen catalog number 10082-147).

[0274] Calcein (AM) (Invitrogen catalog number C3100MP)-labeled MOLP8 target cells and NK92V effector cells were plated at 4x10 4 target cells (MOLP8) and 1.2x10 5Effector cells (NK92V) were seeded into 96-well plates at a density of 0.8×10⁶ cells / mL. The cells were treated with Ab2 at concentrations of 0, 0.001, 0.1, or 1 μg / mL, or with IgG1 control (data not shown) at 37 °C for 1 hour. Calcein fluorescence released from lysed target cells in the supernatant was measured using a multimode plate reader (Envision). The intensity of the fluorescence signal is proportional to the number of lysed target cells.

[0275] Incubating human NK cells with TGF-β for up to 72 h resulted in a time-dependent decrease in Ab2-mediated ADCC (E:T = 3:1). Although NK cells incubated with TGF-β for 24 h retained their lytic activity, the lysis of NK cells incubated for 48 h or 72 h was reduced by approximately 25% - 60% (Figure 2).

[0276] Example 3: TGF-β neutralization restores NK cell lysis activity and Ab2-mediated ADCC

[0277] Logarithmically growing NK92V cells (NK92-05 CD16 V / V; Conkwest) were suspended in growth medium (Middle MyeloCult H5100 (Stemcell Technologies catalog number 05150) supplemented with 100 units / mL human interleukin-2 (R&D catalog number 202-IL-010 / CF)) and seeded into 10-cm culture dishes at a density of 0.8×10⁶ cells / mL. The cells were treated with recombinant human TGF-β at a final concentration of 10 ng / mL, and with 1D11 (murine substitute for Ab1) at a final concentration of 50 μg / mL or isotype control (13C4; data not shown) at 37 °C for 90 hours. 5 Calcein (AM) (Invitrogen catalog number C3100MP)-labeled MOLP8 target cells and NK92V effector cells were seeded into 96-well plates at a density of 4×10⁴ target cells (MOLP8) and 1.2×10⁴ effector cells (NK92V) per well. The cells were treated with 0, 0.001, 0.01, 0.1, or 1 μg / mL of Ab2, or with 1 μg / mL of IgG1 control at 37 °C for 1 hour. Calcein fluorescence released from lysed target cells in the supernatant was measured using a multimode plate reader (Envision). The intensity of the fluorescence signal is proportional to the number of lysed target cells.

[0278] 4 5

[0279] ​​​When TGF-β was neutralized by Ab1, the reduction in NK cell lysis activity induced by TGF-β and Ab2-mediated ADCC was completely restored, but this was not the case with the control antibody (Figure 3).

[0280] Example 4: TGF-β inhibits Ab2-mediated ADCC of NCI-H929 cells, and Ab1 blocks this inhibition

[0281] Human NK cells were isolated from normal human donors (Stem Cell Technologies) by negative selection. The enriched NK cells were cultured in a humidified incubator at 37 °C for 3 days in the presence of IL-2 (at 100 IU / mL), TGF-β (as indicated), and / or Ab1 or isotype control. At this time, exponentially growing NCI-H929 cells were labeled with calcein (AM) and incubated with 1 μg / mL of Ab2 for 30 minutes. Then the NK effector cells were resuspended and combined with the labeled target NCI-H929 cells in a humidified incubator for 1 hour. Ab2-mediated ADCC was quantified by measuring the level of target cells labeled with calcein (AM) using a spectrophotometer. Increasing doses of TGF-β were shown to inhibit the ability of normal human NK cells to kill Ab2-pretreated NCI-H929 cells, where 0.1 ng / mL had little effect on NK cell-mediated cytotoxicity, but 1 and 10 ng / mL blocked ADCC by approximately 50% (Figure 4). Next, ADCC was evaluated at 2 different Ab1 concentrations (100 and 200 μg / mL). Neither the isotype control Ab (IgG4) nor Ab1 had any effect on ADCC when added to the cultures in the absence of TGF-β (Figures 5 and 6). Ab1 was shown to block the ability of TGF-β to inhibit ADCC (Figures 5 and 6, p < 0.005, compared to untreated), while the IgG4 control was unable to block the action of TGF-β to the same extent. Thus, this data demonstrates that Ab1 is capable of alleviating the immunosuppressive effect of TGF-β on NK cell-mediated ADCC.

[0282] Example 5: Neutralization of endogenous TGF-β by Ab1 restores primary NK cell lytic activity and Ab2-mediated ADCC

[0283] JJN3, K562, and RPMI8226 cells were cultured in RPMI 1640 (Invitrogen catalog number 22400-089) supplemented with 10% fetal bovine serum (Invitrogen catalog number 10082-147).

[0284] Human primary NK cells were isolated from normal human PBMCs by negative selection according to the protocol recommended by the manufacturer (StemCell Technologies catalog number 17955RF). The isolated NK cells were cultured alone in a 6-transwell plate or co-cultured with JJN3 cells in the presence or absence of 100 mg / ml Ab1 or isotype control in a 5% CO2 incubator at 37 °C for 90 hours.

[0285] After 90 hours of co-culture, NK cells from the transwell were incubated with calcein (AM) (Invitrogen catalog number C3100MP)-labeled K562 cells at 37 °C for 2 hours for the detection of the cytolytic function of NK cells, or incubated with calcein (AM)-labeled RPMI8226 at 37 °C for 1 hour in the presence of 0.001 or 0.1 mg / ml of Ab2 or 0.1 mg / ml of control Ab2 mutant (Ab*) for the detection of Ab2-mediated ADCC. The calcein fluorescence released from the lysed target cells in the supernatant was measured using a multimode plate reader (Envision). The intensity of the fluorescence signal was proportional to the number of lysed target cells.

[0286] The increased cytolytic activity of primary NK cells (Figure 7) and the increased Ab2-mediated ADCC (Figure 8) demonstrated that the presence of Ab1 neutralized the endogenous TGFβ released by JJN3 cells during the 90-hour co-culture.

[0287] Example 6: Treatment with a combination of Ab1 and Ab2 in patients with multiple myeloma

[0288] The effect of treatment with a combination of antibodies Ab1 and Ab2 can be further evaluated in human patients, such as patients with multiple myeloma who are resistant to one or more other treatments targeting CD38 (e.g., isatuximab or daratumumab). Such patients with multiple myeloma may, for example, be non-responsive to the other treatment targeting CD38 or progress during the treatment with the other treatment targeting CD38.

[0289] The combination of Ab1 and Ab2 is expected to treat multiple myeloma more effectively than Ab2 alone. For example, compared with treatment with Ab2 alone, the combination treatment may result in improved symptoms, reduced bone destruction, enhanced bone formation, leading to bone remodeling and / or bone healing, delayed cancer progression or recurrence, or a longer life expectancy.

[0290] Ab1 and Ab2 can be administered at the following dosages (e.g., 1 - 20 mg / kg) for a certain period of time (e.g., as determined by a doctor) using the following dosing schedules (e.g., weekly, bi - weekly, tri - weekly, or monthly), and the dosing schedule, dosage, and period of time are those required to achieve the desired result. In some embodiments, either or both antibodies can be administered via intravenous infusion. In certain embodiments, during the course of treatment, Ab2 is administered weekly, bi - weekly, tri - weekly, or monthly or any combination thereof at 10 mg / kg or 16 mg / kg of actual body weight.

[0291] Sequence Listing

[0292] SEQ ID NO:1 (Ab1 heavy chain)

[0293]

[0294] SEQ ID NO:2 (Ab1 light chain)

[0295]

[0296] SEQ ID NO:3 (Ab1 heavy chain variable domain)

[0297]

[0298] SEQ ID NO:4 (Ab1 light chain variable domain)

[0299]

[0300] SEQ ID NO:5 (Ab1 heavy chain CDR1)

[0301] SNVIS

[0302] SEQ ID NO:6 (Ab1 heavy chain CDR2)

[0303] GVIPIVDIAN Y

[0304] SEQ ID NO:7 (Ab1 heavy chain CDR3)

[0305] TLGLVLDAMD Y

[0306] SEQ ID NO:8 (Ab1 light chain CDR1)

[0307] RASQSLGSSY LA

[0308] SEQ ID NO:9 (Ab1 light chain CDR2)

[0309] GASSRAP

[0310] SEQ ID NO:10 (Ab1 Light Chain CDR3)

[0311] QQYADSPIT

[0312] SEQ ID NO:11 (Ab2 Heavy Chain)

[0313]

[0314] SEQ ID NO:12 (Ab2 Light Chain)

[0315]

[0316] SEQ ID NO:13 (Ab2 Heavy Chain Variable Domain)

[0317]

[0318] SEQ ID NO:14 (Ab2 Light Chain Variable Domain)

[0319]

[0320] SEQ ID NO:15 (Ab2 Heavy Chain CDR1)

[0321] DYWMQ

[0322] SEQ ID NO:16 (Ab2 Heavy Chain CDR2)

[0323] TIYPGDGDTG YAQKFQG

[0324] SEQ ID NO:17 (Ab2 Heavy Chain CDR3)

[0325] GDYYGSNSLD Y

[0326] SEQ ID NO:18 (Ab2 Light Chain CDR1)

[0327] KASQDVSTVV A

[0328] SEQ ID NO:19 (Ab2 Light Chain CDR2)

[0329] SASYRYI

[0330] SEQ ID NO:20 (Ab2 Light Chain CDR3)

[0331] QQHYSPPYT

[0332] SEQ ID NO:21 (Human TGF-β1): SwissProt P01137

[0333]

[0334] SEQ ID NO:22 (Human TGF-β2): SwissProt P08112

[0335]

[0336] SEQ ID NO:23 (Human TGF-β3): SwissProt P10600

[0337]

[0338] SEQ ID NO:24 (Human CD38): GenBank NP_001766

[0339]

[0340] SEQ ID NO:25 (Ab3 heavy chain)

[0341]

[0342]

[0343] SEQ ID NO:26 (Ab3 light chain)

[0344]

[0345] SEQ ID NO:27 (Ab3 heavy chain variable domain)

[0346]

[0347] SEQ ID NO:28 (Ab3 light chain variable domain)

[0348]

[0349] SEQ ID NO:29 (Ab3 heavy chain CDR1)

[0350] GYTFTSYA

[0351] SEQ ID NO:30 (Ab3 heavy chain CDR2)

[0352] IYPGQGGT

[0353] SEQ ID NO:31 (Ab3 heavy chain CDR3)

[0354] ARTGGLRRAY FTY

[0355] SEQ ID NO:32 (Ab3 light chain CDR1)

[0356] QSVSSYGQGF

[0357] SEQ ID NO:33 (Ab3 light chain CDR2)

[0358] GAS

[0359] SEQ ID NO:34 (Ab3 light chain CDR3)

[0360] QQNKEDPWT

[0361] SEQ ID NO:35 (Ab4 heavy chain)

[0362]

[0363] SEQ ID NO:36 (Ab4 light chain)

[0364]

[0365] SEQ ID NO:37 (Ab4 heavy chain variable domain)

[0366]

[0367] SEQ ID NO:38 (Ab4 light chain variable domain)

[0368]

[0369] SEQ ID NO:39 (Ab4 heavy chain CDR1)

[0370] GFTFSSYG

[0371] SEQ ID NO:40 (Ab4 heavy chain CDR2)

[0372] IWYDGSNK

[0373] SEQ ID NO:41 (Ab4 heavy chain CDR3)

[0374] ARMFRGAFDY

[0375] SEQ ID NO:42 (Ab4 light chain CDR1)

[0376] QGIRND

[0377] SEQ ID NO:43 (Ab4 light chain CDR2)

[0378] AAS

[0379] SEQ ID NO:44 (Ab4 light chain CDR3)

[0380] LQDYIYYPT。 Sequence Listing <110> Sanofi <120> Anti-cancer combination therapy targeting CD38 and TGF-β <130> 022548.WO025 <140> <141> <150> 62 / 696,198 <151> 2018-07-10 <160> 44 <170> PatentIn version 3.5 <210> 1 <211> 447 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note = "Description of artificial sequence: synthetic polypeptide" <400> 1 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn 20 25 30 Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro 210 215 220 Pro Cys Pro 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> 2 <211> 215 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / note="Description of artificial sequence: Synthetic polypeptide" <400> 2 Glu Thr Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Leu Gly Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ala Asp Ser Pro 85 90 95 Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 3 <211> 120 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Comment="Description of artificial sequence: synthetic polypeptide" <400> 3 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn 20 25 30 Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 4 <211> 108 <212> PRT <213> artificial sequence <220> <221> source <223> / Comment="Description of artificial sequence: synthetic polypeptide" <400> 4 Glu Thr Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Leu Gly Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Ala Asp Ser Pro 85 90 95 Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 <210> 5 <211> 5 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic peptide" <400> 5 Ser Asn Val Ile Ser 1 5 <210> 6 <211> 11 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 6 Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr 1 5 10 <210> 7 <211> 11 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 7 Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr 1 5 10 <210> 8 <211> 12 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 8 Arg Ala Ser Gln Ser Leu Gly Ser Ser Tyr Leu Ala 1 5 10 <210> 9 <211> 7 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 9 Gly Ala Ser Ser Arg Ala Pro 1 5 <210> 10 <211> 9 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic peptide" <400> 10 Gln Gln Tyr Ala Asp Ser Pro Ile Thr 1 5 <210> 11 <211> 450 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic polypeptide" <400> 11 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Ala Lys Pro Gly Thr 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Trp Met Gln Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Thr Ile Tyr Pro Gly Asp Gly Asp Thr Gly Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Lys Thr Val Tyr 65 70 75 80 Met His Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Tyr Gly Ser Asn Ser Leu Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser 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 Lys 450 <210> 12 <211> 214 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic polypeptide" <400> 12 Asp Ile Val Met Thr Gln Ser His Leu Ser Met Ser Thr Ser Leu Gly 1 5 10 15 Asp Pro Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Val 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Arg Arg Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Ile Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ala Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu 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> 13 <211> 120 <212> PRT <213> Artificial sequence <220> <221> Source <223> / note="Description of artificial sequence: Synthetic polypeptide" <400> 13 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Ala Lys Pro Gly Thr 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Trp Met Gln Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Thr Ile Tyr Pro Gly Asp Gly Asp Thr Gly Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Lys Thr Val Tyr 65 70 75 80 Met His Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Asp Tyr Tyr Gly Ser Asn Ser Leu Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 14 <211> 107 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Comment="Description of artificial sequence: Synthetic polypeptide" <400> 14 Asp Ile Val Met Thr Gln Ser His Leu Ser Met Ser Thr Ser Leu Gly 1 5 10 15 Asp Pro Val Ser Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Thr Val 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Arg Arg Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Ile Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ala Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Ser Pro Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 15 <211> 5 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Comment="Description of artificial sequence: Synthetic peptide" <400> 15 Asp Tyr Trp Met Gln 1 5 <210> 16 <211> 17 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 16 Thr Ile Tyr Pro Gly Asp Gly Asp Thr Gly Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <210> 17 <211> 11 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 17 Gly Asp Tyr Tyr Gly Ser Asn Ser Leu Asp Tyr 1 5 10 <210> 18 <211> 11 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 18 Lys Ala Ser Gln Asp Val Ser Thr Val Val Ala 1 5 10 <210> 19 <211> 7 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Annotation="Description of artificial sequence: Synthetic peptide" <400> 19 Ser Ala Ser Tyr Arg Tyr Ile 1 5 <210> 20 <211> 9 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Annotation="Description of artificial sequence: Synthetic peptide" <400> 20 Gln Gln His Tyr Ser Pro Pro Tyr Thr 1 5 <210> 21 <211> 390 <212> PRT <213> Homo sapiens <400> 21 Met Pro Pro Ser Gly Leu Arg Leu Leu Leu Leu Leu Leu Pro Leu Leu 1 5 10 15 Trp Leu Leu Val Leu Thr Pro Gly Arg Pro Ala Ala Gly Leu Ser Thr 20 25 30 Cys Lys Thr Ile Asp Met Glu Leu Val Lys Arg Lys Arg Ile Glu Ala 35 40 45 Ile Arg Gly Gln Ile Leu Ser Lys Leu Arg Leu Ala Ser Pro Pro Ser 50 55 60 Gln Gly Glu Val Pro Pro Gly Pro Leu Pro Glu Ala Val Leu Ala Leu 65 70 75 80 Tyr Asn Ser Thr Arg Asp Arg Val Ala Gly Glu Ser Ala Glu Pro Glu 85 90 95 Pro Glu Pro Glu Ala Asp Tyr Tyr Ala Lys Glu Val Thr Arg Val Leu 100 105 110 Met Val Glu Thr His Asn Glu Ile Tyr Asp Lys Phe Lys Gln Ser Thr 115 120 125 His Ser Ile Tyr Met Phe Phe Asn Thr Ser Glu Leu Arg Glu Ala Val 130 135 140 Pro Glu Pro Val Leu Leu Ser Arg Ala Glu Leu Arg Leu Leu Arg Leu 145 150 155 160 Lys Leu Lys Val Glu Gln His Val Glu Leu Tyr Gln Lys Tyr Ser Asn 165 170 175 Asn Ser Trp Arg Tyr Leu Ser Asn Arg Leu Leu Ala Pro Ser Asp Ser 180 185 190 Pro Glu Trp Leu Ser Phe Asp Val Thr Gly Val Val Arg Gln Trp Leu 195 200 205 Ser Arg Gly Gly Glu Ile Glu Gly Phe Arg Leu Ser Ala His Cys Ser 210 215 220 Cys Asp Ser Arg Asp Asn Thr Leu Gln Val Asp Ile Asn Gly Phe Thr 225 230 235 240 Thr Gly Arg Arg Gly Asp Leu Ala Thr Ile His Gly Met Asn Arg Pro 245 250 255 Phe Leu Leu Leu Met Ala Thr Pro Leu Glu Arg Ala Gln His Leu Gln 260 265 270 Ser Ser Arg His Arg Arg Ala Leu Asp Thr Asn Tyr Cys Phe Ser Ser 275 280 285 Thr Glu Lys Asn Cys Cys Val Arg Gln Leu Tyr Ile Asp Phe Arg Lys 290 295 300 Asp Leu Gly Trp Lys Trp Ile His Glu Pro Lys Gly Tyr His Ala Asn 305 310 315 320 Phe Cys Leu Gly Pro Cys Pro Tyr Ile Trp Ser Leu Asp Thr Gln Tyr 325 330 335 Ser Lys Val Leu Ala Leu Tyr Asn Gln His Asn Pro Gly Ala Ser Ala 340 345 350 Ala Pro Cys Cys Val Pro Gln Ala Leu Glu Pro Leu Pro Ile Val Tyr 355 360 365 Tyr Val Gly Arg Lys Pro Lys Val Glu Gln Leu Ser Asn Met Ile Val 370 375 380 Arg Ser Cys Lys Cys Ser 385 390 <210> 22 <211> 414 <212> PRT <213> Homo sapiens <400> 22 Met His Tyr Cys Val Leu Ser Ala Phe Leu Ile Leu His Leu Val Thr 1 5 10 15 Val Ala Leu Ser Leu Ser Thr Cys Ser Thr Leu Asp Met Asp Gln Phe 20 25 30 Met Arg Lys Arg Ile Glu Ala Ile Arg Gly Gln Ile Leu Ser Lys Leu 35 40 45 Lys Leu Thr Ser Pro Pro Glu Asp Tyr Pro Glu Pro Glu Glu Val Pro 50 55 60 Pro Glu Val Ile Ser Ile Tyr Asn Ser Thr Arg Asp Leu Leu Gln Glu 65 70 75 80 Lys Ala Ser Arg Arg Ala Ala Ala Cys Glu Arg Glu Arg Ser Asp Glu 85 90 95 Glu Tyr Tyr Ala Lys Glu Val Tyr Lys Ile Asp Met Pro Pro Phe Phe 100 105 110 Pro Ser Glu Asn Ala Ile Pro Pro Thr Phe Tyr Arg Pro Tyr Phe Arg 115 120 125 Ile Val Arg Phe Asp Val Ser Ala Met Glu Lys Asn Ala Ser Asn Leu 130 135 140 Val Lys Ala Glu Phe Arg Val Phe Arg Leu Gln Asn Pro Lys Ala Arg 145 150 155 160 Val Pro Glu Gln Arg Ile Glu Leu Tyr Gln Ile Leu Lys Ser Lys Asp 165 170 175 Leu Thr Ser Pro Thr Gln Arg Tyr Ile Asp Ser Lys Val Val Lys Thr 180 185 190 Arg Ala Glu Gly Glu Trp Leu Ser Phe Asp Val Thr Asp Ala Val His 195 200 205 Glu Trp Leu His His Lys Asp Arg Asn Leu Gly Phe Lys Ile Ser Leu 210 215 220 His Cys Pro Cys Cys Thr Phe Val Pro Ser Asn Asn Tyr Ile Ile Pro 225 230 235 240 Asn Lys Ser Glu Glu Leu Glu Ala Arg Phe Ala Gly Ile Asp Gly Thr 245 250 255 Ser Thr Tyr Thr Ser Gly Asp Gln Lys Thr Ile Lys Ser Thr Arg Lys 260 265 270 Lys Asn Ser Gly Lys Thr Pro His Leu Leu Leu Met Leu Leu Pro Ser 275 280 285 Tyr Arg Leu Glu Ser Gln Gln Thr Asn Arg Arg Lys Lys Arg Ala Leu 290 295 300 Asp Ala Ala Tyr Cys Phe Arg Asn Val Gln Asp Asn Cys Cys Leu Arg 305 310 315 320 Pro Leu Tyr Ile Asp Phe Lys Arg Asp Leu Gly Trp Lys Trp Ile His 325 330 335 Glu Pro Lys Gly Tyr Asn Ala Asn Phe Cys Ala Gly Ala Cys Pro Tyr 340 345 350 Leu Trp Ser Ser Asp Thr Gln His Ser Arg Val Leu Ser Leu Tyr Asn 355 360 365 Thr Ile Asn Pro Glu Ala Ser Ala Ser Pro Cys Cys Val Ser Gln Asp 370 375 380 Leu Glu Pro Leu Thr Ile Leu Tyr Tyr Ile Gly Lys Thr Pro Lys Ile 385 390 395 400 Glu Gln Leu Ser Asn Met Ile Val Lys Ser Cys Lys Cys Ser 405 410 <210> 23 <211> 412 <212> PRT <213> Homo sapiens <400> 23 Met Lys Met His Leu Gln Arg Ala Leu Val Val Leu Ala Leu Leu Asn 1 5 10 15 Phe Ala Thr Val Ser Leu Ser Leu Ser Thr Cys Thr Thr Leu Asp Phe 20 25 30 Gly His Ile Lys Lys Lys Arg Val Glu Ala Ile Arg Gly Gln Ile Leu 35 40 45 Ser Lys Leu Arg Leu Thr Ser Pro Pro Glu Pro Thr Val Met Thr His 50 55 60 Val Pro Tyr Gln Val Leu Ala Leu Tyr Asn Ser Thr Arg Glu Leu Leu 65 70 75 80 Glu Glu Met His Gly Glu Arg Glu Glu Gly Cys Thr Gln Glu Asn Thr 85 90 95 Glu Ser Glu Tyr Tyr Ala Lys Glu Ile His Lys Phe Asp Met Ile Gln 100 105 110 Gly Leu Ala Glu His Asn Glu Leu Ala Val Cys Pro Lys Gly Ile Thr 115 120 125 Ser Lys Val Phe Arg Phe Asn Val Ser Ser Val Glu Lys Asn Arg Thr 130 135 140 Asn Leu Phe Arg Ala Glu Phe Arg Val Leu Arg Val Pro Asn Pro Ser 145 150 155 160 Ser Lys Arg Asn Glu Gln Arg Ile Glu Leu Phe Gln Ile Leu Arg Pro 165 170 175 Asp Glu His Ile Ala Lys Gln Arg Tyr Ile Gly Gly Lys Asn Leu Pro 180 185 190 Thr Arg Gly Thr Ala Glu Trp Leu Ser Phe Asp Val Thr Asp Thr Val 195 200 205 Arg Glu Trp Leu Leu Arg Arg Glu Ser Asn Leu Gly Leu Glu Ile Ser 210 215 220 Ile His Cys Pro Cys His Thr Phe Gln Pro Asn Gly Asp Ile Leu Glu 225 230 235 240 Asn Ile His Glu Val Met Glu Ile Lys Phe Lys Gly Val Asp Asn Glu 245 250 255 Asp Asp His Gly Arg Gly Asp Leu Gly Arg Leu Lys Lys Gln Lys Asp 260 265 270 His His Asn Pro His Leu Ile Leu Met Met Ile Pro Pro His Arg Leu 275 280 285 Asp Asn Pro Gly Gln Gly Gly Gln Arg Lys Lys Arg Ala Leu Asp Thr 290 295 300 Asn Tyr Cys Phe Arg Asn Leu Glu Glu Asn Cys Cys Val Arg Pro Leu 305 310 315 320 Tyr Ile Asp Phe Arg Gln Asp Leu Gly Trp Lys Trp Val His Glu Pro 325 330 335 Lys Gly Tyr Tyr Ala Asn Phe Cys Ser Gly Pro Cys Pro Tyr Leu Arg 340 345 350 Ser Ala Asp Thr Thr His Ser Thr Val Leu Gly Leu Tyr Asn Thr Leu 355 360 365 Asn Pro Glu Ala Ser Ala Ser Pro Cys Cys Val Pro Gln Asp Leu Glu 370 375 380 Pro Leu Thr Ile Leu Tyr Tyr Val Gly Arg Thr Pro Lys Val Glu Gln 385 390 395 400 Leu Ser Asn Met Val Val Lys Ser Cys Lys Cys Ser 405 410 <210> 24 <211> 300 <212> PRT <213> Homo sapiens <400> 24 Met Ala Asn Cys Glu Phe Ser Pro Val Ser Gly Asp Lys Pro Cys Cys 1 5 10 15 Arg Leu Ser Arg Arg Ala Gln Leu Cys Leu Gly Val Ser Ile Leu Val 20 25 30 Leu Ile Leu Val Val Val Leu Ala Val Val Val Pro Arg Trp Arg Gln 35 40 45 Gln Trp Ser Gly Pro Gly Thr Thr Lys Arg Phe Pro Glu Thr Val Leu 50 55 60 Ala Arg Cys Val Lys Tyr Thr Glu Ile His Pro Glu Met Arg His Val 65 70 75 80 Asp Cys Gln Ser Val Trp Asp Ala Phe Lys Gly Ala Phe Ile Ser Lys 85 90 95 His Pro Cys Asn Ile Thr Glu Glu Asp Tyr Gln Pro Leu Met Lys Leu 100 105 110 Gly Thr Gln Thr Val Pro Cys Asn Lys Ile Leu Leu Trp Ser Arg Ile 115 120 125 Lys Asp Leu Ala His Gln Phe Thr Gln Val Gln Arg Asp Met Phe Thr 130 135 140 Leu Glu Asp Thr Leu Leu Gly Tyr Leu Ala Asp Asp Leu Thr Trp Cys 145 150 155 160 Gly Glu Phe Asn Thr Ser Lys Ile Asn Tyr Gln Ser Cys Pro Asp Trp 165 170 175 Arg Lys Asp Cys Ser Asn Asn Pro Val Ser Val Phe Trp Lys Thr Val 180 185 190 Ser Arg Arg Phe Ala Glu Ala Ala Cys Asp Val Val His Val Met Leu 195 200 205 Asn Gly Ser Arg Ser Lys Ile Phe Asp Lys Asn Ser Thr Phe Gly Ser 210 215 220 Val Glu Val His Asn Leu Gln Pro Glu Lys Val Gln Thr Leu Glu Ala 225 230 235 240 Trp Val Ile His Gly Gly Arg Glu Asp Ser Arg Asp Leu Cys Gln Asp 245 250 255 Pro Thr Ile Lys Glu Leu Glu Ser Ile Ile Ser Lys Arg Asn Ile Gln 260 265 270 Phe Ser Cys Lys Asn Ile Tyr Arg Pro Asp Lys Phe Leu Gln Cys Val 275 280 285 Lys Asn Pro Glu Asp Ser Ser Cys Thr Ser Glu Ile 290 295 300 <210> 25 <211> 449 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic polypeptide" <400> 25 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Met His Trp Val Lys Glu Ala Pro Gly Gln Arg Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Tyr Pro Gly Gln Gly Gly Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Ala Thr Leu Thr Ala Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Arg Thr Gly Gly Leu Arg Arg Ala Tyr Phe Thr 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 Ala Gly 225 230 235 240 Pro Asp 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 Leu Pro Glu 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 <210> 26 <211> 218 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / note="Description of artificial sequence: Synthetic polypeptide" <400> 26 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Ile Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 20 25 30 Gly Gln Gly Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile 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 Pro Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Asn Lys 85 90 95 Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu 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> 27 <211> 120 <212> PRT <213> Artificial sequence <220> <221> Source <223> / note="Description of artificial sequence: Synthetic polypeptide" <400> 27 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Ala Met His Trp Val Lys Glu Ala Pro Gly Gln Arg Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Tyr Pro Gly Gln Gly Gly Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Ala Thr Leu Thr Ala Asp Thr Ser Ala Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Arg Thr Gly Gly Leu Arg Arg Ala Tyr Phe Thr Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 28 <211> 111 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic polypeptide" <400> 28 Asp Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Ile Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 20 25 30 Gly Gln Gly Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile 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 Pro Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Asn Lys 85 90 95 Glu Asp Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 29 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic peptide" <400> 29 Gly Tyr Thr Phe Thr Ser Tyr Ala 1 5 <210> 30 <211> 8 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: synthetic peptide" <400> 30 Ile Tyr Pro Gly Gln Gly Gly Thr 1 5 <210> 31 <211> 13 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: synthetic peptide" <400> 31 Ala Arg Thr Gly Gly Leu Arg Arg Ala Tyr Phe Thr Tyr 1 5 10 <210> 32 <211> 10 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: synthetic peptide" <400> 32 Gln Ser Val Ser Ser Tyr Gly Gln Gly Phe 1 5 10 <210> 33 <211> 3 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic peptide" <400> 33 Gly Ala Ser 1 <210> 34 <211> 9 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic peptide" <400> 34 Gln Gln Asn Lys Glu Asp Pro Trp Thr 1 5 <210> 35 <211> 446 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic polypeptide" <400> 35 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 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 Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gly 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 Met Phe Arg Gly Ala Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Ala Gly Pro Asp 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 His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr 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 Ala Leu Pro Leu Pro Glu 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 Arg Asp Glu Leu 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 Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln 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 Pro Gly 435 440 445 <210> 36 <211> 214 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic polypeptide" <400> 36 Ala 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 Gly Ile Arg Asn Asp 20 25 30 Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Gly Leu Gln Pro 65 70 75 80 Glu Asp Ser Ala Thr Tyr Tyr Cys Leu Gln Asp Tyr Ile Tyr Tyr Pro 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> 37 <211> 117 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note="Description of artificial sequence: Synthetic polypeptide" <400> 37 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 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 Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gly 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 Met Phe Arg Gly Ala Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 38 <211> 107 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Comment="Description of artificial sequence: Synthetic polypeptide" <400> 38 Ala 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 Gly Ile Arg Asn Asp 20 25 30 Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Gly Leu Gln Pro 65 70 75 80 Glu Asp Ser Ala Thr Tyr Tyr Cys Leu Gln Asp Tyr Ile Tyr Tyr Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 39 <211> 8 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Comment="Description of artificial sequence: Synthetic peptide" <400> 39 Gly Phe Thr Phe Ser Ser Tyr Gly 1 5 <210> 40 <211> 8 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 40 Ile Trp Tyr Asp Gly Ser Asn Lys 1 5 <210> 41 <211> 10 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 41 Ala Arg Met Phe Arg Gly Ala Phe Asp Tyr 1 5 10 <210> 42 <211> 6 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 42 Gln Gly Ile Arg Asn Asp 1 5 <210> 43 <211> 3 <212> PRT <213> Artificial sequence <220> <221> Source <223> / Note = "Description of artificial sequence: Synthetic peptide" <400> 43 Ala Ala Ser 1 <210> 44 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> Source <223> / note="Description of artificial sequence: Synthetic peptide" <400> 44 Leu Gln Asp Tyr Ile Tyr Tyr Pro Thr 1 5

Claims

1. Use of an anti-CD38 antibody in the preparation of a medicament for the combined treatment of multiple myeloma in a human patient in need thereof with an anti-TGF-β antibody or an antigen-binding fragment thereof, wherein the anti-CD38 antibody has heavy chain CDR1 (HCDR1), HCDR2, HCDR3, light chain CDR1 (LCDR1), LCDR2 and LCDR3 respectively consisting of the amino acid sequences of SEQ ID NOs: 15-20, and the anti-TGF-β antibody or an antigen-binding fragment thereof has HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 respectively consisting of the amino acid sequences of SEQ ID NOs: 5-10, and the anti-CD38 antibody has a human IgG1 Fc region, and the anti-TGF-β antibody has a human IgG4 Fc region.

2. Use of an anti-TGF-β antibody or an antigen-binding fragment thereof in the preparation of a medicament for the combined treatment of multiple myeloma in a human patient in need thereof with an anti-CD38 antibody, wherein the anti-CD38 antibody has heavy chain CDR1 (HCDR1), HCDR2, HCDR3, light chain CDR1 (LCDR1), LCDR2 and LCDR3 respectively consisting of the amino acid sequences of SEQ ID NOs: 15-20, and the anti-TGF-β antibody or an antigen-binding fragment thereof has HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 respectively consisting of the amino acid sequences of SEQ ID NOs: 5-10, and the anti-CD38 antibody has a human IgG1 Fc region, and the anti-TGF-β antibody has a human IgG4 Fc region.

3. Use according to claim 1 or 2, wherein the anti-CD38 antibody has a heavy chain variable domain (V H ) and a light chain variable domain (V L ) containing the amino acid sequences set forth in SEQ ID NO: 13 and 14, respectively; and wherein the anti-TGF-β antibody has V H and V L containing the amino acid sequences set forth in SEQ ID NO: 3 and 4, respectively.

4. The use according to claim 3, wherein the anti-CD38 antibody has a heavy chain (HC) and a light chain (LC) respectively comprising the amino acid sequences of SEQ ID NOs: 11 and 12; and wherein the anti-TGF-β antibody has a HC and a LC respectively comprising the amino acid sequences of SEQ ID NOs: 1 and 2.

5. Use of an anti-CD38 antibody in the preparation of a medicament for the combined treatment of multiple myeloma in human patients in need thereof with an anti-TGF-β antibody, wherein the anti-CD38 antibody has a heavy chain variable domain (V H ) and a light chain variable domain (V L ) respectively comprising the amino acid sequences of SEQ ID NO: 13 and 14, and a human IgG1 Fc region; or having a heavy chain (HC) and a light chain (LC) respectively comprising the amino acid sequences of SEQ ID NOs: 11 and 12; and The anti-TGF-β antibody has V that respectively comprises the amino acid sequences of SEQ ID NO:3 and 4 H and V L , and a human IgG4 Fc region; or having a HC and a LC respectively comprising the amino acid sequences of SEQ ID NOs: 1 and 2.

6. Use of an anti-TGF-β antibody for the manufacture of a medicament for the combined treatment of multiple myeloma in a human patient in need thereof with an anti-CD38 antibody, wherein the anti-TGF-β antibody V having amino acid sequences comprising SEQ ID NO:3 and 4, respectively H and V L ; or has a HC and a LC respectively comprising the amino acid sequences of SEQ ID NOs: 1 and 2, and has a human IgG4 Fc region; and wherein the anti-CD38 antibody A heavy chain variable domain (V H ) and a light chain variable domain (V L ) having amino acid sequences respectively containing SEQ ID NO: 13 and 14, and a human IgG1 Fc region; or has a heavy chain (HC) and a light chain (LC) respectively comprising the amino acid sequences of SEQ ID NOs: 11 and 12.

7. The use according to claim 1 or 2, wherein the anti-CD38 antibody and the anti-TGF-β antibody are administered to the patient sequentially.

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