CD38 antibodies for treatment of multiple myeloma

Abstract

Isolated human monoclonal antibodies and related antibody-based compositions and molecules that bind to human CD38 are described. Pharmaceutical compositions comprising the human antibodies and therapeutic and diagnostic methods of using the human antibodies are also described.

Description

[0001] This application is a divisional application of the invention patent application with application number 202011391749.5, filed on March 23, 2006, entitled “CD38 Antibody for Treating Multiple Myeloma.” The Chinese invention patent application with application number 202011391749.5 is a divisional application of the invention patent application with application number 201710164800.0. The Chinese invention patent application with application number 201710164800.0 is a divisional application of the invention patent application with application number 200680017915.7. Technical Field

[0002] The present invention relates to antibodies that bind to CD38, have specific characteristics, and are furthermore useful for treating multiple myeloma. Background Art

[0003] Multiple myeloma is a B-cell malignancy characterized by the latent accumulation of secretory plasma cells with a low proliferation rate and prolonged life span in the bone marrow. The disease ultimately attacks the bones and bone marrow, causing multiple tumors and malignancies throughout the skeletal system.

[0004] Multiple myeloma (MM) accounts for approximately 1% of all cancers and slightly more than 10% of all hematological malignancies. The incidence of MM increases in the elderly, with an average age of diagnosis of approximately 61 years.

[0005] Currently available treatments for multiple myeloma include chemotherapy, stem cell transplantation, (thalidomide), (bortezomib), (pamidronate) and (Zolephosphamide). Current treatment regimens, which include combinations of chemotherapy agents such as vincristine, BCNU, melphalan, cyclophosphamide, daunorubicin, and prednisone or dexamethasone, result in complete remission rates of only approximately 5%, with a median survival of approximately 36-48 months from the time of diagnosis. Recent advances in the use of high-dose chemotherapy followed by autologous bone marrow or peripheral blood mononuclear cell transplantation have improved complete remission rates and duration of remission. However, overall survival has been only slightly prolonged, and there is no evidence of a cure. Ultimately, all MM patients will relapse, even with maintenance therapy with interferon-α (IFN-α) alone or in combination with steroids.

[0006] The efficacy of available MM chemotherapy is limited by the low cell proliferation rate and the development of multidrug resistance. For over 90% of MM patients, the disease is resistant to chemotherapy. Therefore, alternative treatment options are exploring immunotherapies that target surface antigens on plasma cells.

[0007] CD38 is an example of an antigen expressed on such malignant plasma cells and is expressed in a variety of hematological malignancies, including but not limited to multiple myeloma, B-cell chronic lymphocytic leukemia, B-cell acute lymphocytic leukemia, Waldenstrom's macroglobulinemia, primary systemic amyloidosis, mantle cell lymphoma, prolymphocytic / granulocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, follicular lymphoma, NK cell leukemia, and plasma cell leukemia. CD38 expression has been described in epithelial / endothelial cells of various origins, including glandular epithelial cells of the prostate, pancreatic islet cells, bile duct epithelial cells of glands including the parotid gland, bronchial epithelial cells, cells in the testis and ovary, and cancerous epithelial cells in colorectal adenocarcinoma. Diseases in which CD38 expression is involved include, but are not limited to, bronchial epithelial carcinoma of the lung, breast cancer (malignant proliferation of the epithelial surface of the ducts or lobules of the breast), pancreatic tumors (islet tumors) derived from B cells, and tumors derived from intestinal epithelium (e.g., adenocarcinoma and squamous cell carcinoma). In the CNS, neuroblastoma expresses CD38. Other diseases include prostate cancer, seminoma in testicular and ovarian cancer.

[0008] Normally, CD38 is expressed by hematopoietic cells in solid tissues. Among hematopoietic cells, most medullary thymocytes are CD38 + , resting and circulating T and B cells are CD38 - , the activated cells are CD38 + CD38 is also expressed in about 80% of dormant NK cells and monocytes, as well as in lymphoblasts, plasma B cells, and some follicular cells in the germinal centers of lymph nodes. CD38 is also expressed by dendritic cells. Most normal bone marrow cells, especially precursor cells, express CD38. In addition, 50-80% of umbilical cord blood cells are CD38. + , and can survive in human blood for 2 to 3 years. In addition to lymphoid precursor cells, CD38 is also expressed on red blood cells and platelets.

[0009] With respect to solid tissues, CD38 is expressed by intraepithelial cells and lamina propria lymphocytes in the gut, by Purkinje cells and neurofibrillary tangles in the brain, by epithelial cells in the prostate, by beta cells in the pancreas, by osteoclasts in bone, by retinal cells in the eye, and by the sarcolemma in smooth and striated muscle.

[0010] The functions of CD38 include receptor mediation in adhesion and signal transduction events and extracellular enzyme activity. As an extracellular enzyme, CD38 uses NAD +As substrates, it forms cyclic ADP-ribose (cADPR) and ADPR, as well as nicotine and nicotinic acid adenine dinucleotide phosphate (NAADP). It has been shown that cADPR can act as a Ca 2+ Secondary messengers that migrate from the endoplasmic reticulum. In addition to Ca 2+ In addition to signaling through IgG1, CD38 signaling also occurs through the interaction of antigens with receptor complexes on T and B cells or other types of complexes such as MHC molecules, and in this way participates in several cellular responses and can initiate the conversion and secretion of IgG1.

[0011] Anti-CD38 antibodies have been reported in the literature, for example, by Lande R, et al., Cell Immunol. 220(1), 30-8 (2002), Ausiello CM, et al., Tissue Antigens. 56(6), 539-47 (2000), and Cotner T, et al., Int J Immunopharmacol. 3(3), 255-68 (1981). CD38 has multiple functions and may or may not be activated by molecules that bind to CD38. For example, the mouse anti-CD38 antibody IB4 has antagonistic properties against CD38. IB4 has been shown to induce T cell activation in Jurkat cells (Zubiaur M, et al., J Immunol. 159(1), 193-205 (1997), to induce significant proliferation of peripheral blood mononuclear cells (PBMCs), to induce release of significant IL-6 levels and to induce release of detectable IFN-γ levels (Lande, Zubiaur Morra, Ansiello, supra). Summary of the Invention

[0012] The present invention provides an antibody that binds to human CD38 encoded by:

[0013] (i) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 1 and SEQ ID No: 6, respectively, in their variable regions;

[0014] (ii) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 11 and SEQ ID No: 16, respectively, in their variable regions;

[0015] (iii) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 21 and SEQ ID No: 26, respectively, in their variable regions; or

[0016] (iv) Human light chain and human heavy chain nucleic acids containing nucleotide sequences in their variable regions that contain conservative sequence changes listed in (i), (ii) or (iii).

[0017] The present invention provides a V comprising a sequence as shown in SEQ ID No: 10. H Antibodies that bind to human CD38.

[0018] The present invention provides a V comprising a sequence as shown in SEQ ID No: 5. L The sequence of CDR3 and V having the sequence in SEQ ID No: 10 H Antibodies that bind to human CD38.

[0019] The present invention provides an antibody that binds to human CD38, comprising a human light chain and a human heavy chain variable region, wherein the light chain variable region comprises a V having a sequence as shown in SEQ ID No: 3. L CDR1, V having the sequence in SEQ ID No: 4 L CDR2 and V having the sequence as in SEQ ID No: 5 L CDR3, the heavy chain variable region comprises a V having a sequence as in SEQ ID No: 8 H CDR1, V having the sequence in SEQ ID No: 9 H CDR2 and V having the sequence in SEQ ID No: 10 H CDR3.

[0020] The present invention provides an antibody that binds to human CD38, which comprises a V having an amino acid sequence as shown in SEQ ID No: 2. L district.

[0021] The present invention provides an antibody that binds to human CD38, comprising a V sequence having at least about 90%, such as at least about 95%, amino acid sequence identity to the sequence in SEQ ID No: 2. L district.

[0022] The present invention provides an antibody that binds to human CD38, comprising a V having an amino acid sequence as shown in SEQ ID No: 7. H district.

[0023] The present invention provides an antibody that binds to human CD38, comprising a V sequence spanning SEQ ID No: 7.H CDR1-V H The amino acid sequence of the CDR3 region is V H district.

[0024] The present invention provides an antibody that binds to human CD38, which comprises a sequence as in SEQ ID No: 7 or a sequence spanning V as in SEQ ID No: 7. H CDR1-V H The region of CDR3 has at least about 90%, such as at least about 95%, amino acid sequence identity to V H district.

[0025] The present invention provides an antibody that binds to human CD38, which comprises a sequence as in SEQ ID No: 7 or a sequence spanning V as in SEQ ID No: 7. H CDR1-V H The CDR3 region has 1-5, for example 1-3 amino acid substitutions, deletions or additions. H district.

[0026] The present invention provides an antibody that binds to human CD38, which comprises V as defined above. L region and V as defined above H district.

[0027] The present invention provides an antibody that binds to human CD38, comprising a V having a sequence as shown in SEQ ID No: 20. H CDR3.

[0028] The present invention provides an antibody that binds to human CD38, comprising a V having a sequence as shown in SEQ ID No: 15. L CDR3 and V having the sequence in SEQ ID No: 20 H CDR3.

[0029] The present invention provides an antibody that binds to human CD38, comprising a human light chain and a human heavy chain variable region, wherein the light chain variable region comprises a V having a sequence as shown in SEQ ID No: 13. L CDR1, V having the sequence in SEQ ID No: 14 L CDR2 and V having the sequence in SEQ ID No: 15 L CDR3, the heavy chain variable region comprises a V having a sequence as in SEQ ID No: 18 H CDR1, V having the sequence in SEQ ID No: 19 H CDR2 and V having the sequence in SEQ ID No: 20 H CDR3.

[0030] The present invention provides an antibody that binds to human CD38, which comprises a V having an amino acid sequence as shown in SEQ ID No: 12. L district.

[0031] The present invention provides an antibody that binds to human CD38, comprising a V sequence having at least about 90%, such as at least about 95%, amino acid sequence identity to the sequence in SEQ ID No: 12. L district.

[0032] The present invention provides an antibody that binds to human CD38, which comprises a V having an amino acid sequence as shown in SEQ ID No: 17. H district.

[0033] The present invention provides an antibody that binds to human CD38, comprising a V sequence spanning SEQ ID No: 17. H CDR1-V H The amino acid sequence of the CDR3 region is V H district.

[0034] The present invention provides an antibody that binds to human CD38, which comprises a sequence as in SEQ ID No: 17 or a sequence spanning V as in SEQ ID No: 17. H CDR1-V H The region of CDR3 has at least about 90%, such as at least about 95%, amino acid sequence identity to V H district.

[0035] The present invention provides an antibody that binds to human CD38, which comprises a sequence as in SEQ ID No: 17 or a sequence spanning V as in SEQ ID No: 17. H CDR1-V H The CDR3 region has 1-5, for example 1-3 amino acid substitutions, deletions or additions. H district.

[0036] The present invention provides an antibody that binds to human CD38, which comprises V as defined above. L region and V as defined above H district.

[0037] The present invention provides an antibody that binds to human CD38, comprising a V having a sequence as shown in SEQ ID No: 30. H CDR3.

[0038] The present invention provides an antibody that binds to human CD38, comprising a V having a sequence as shown in SEQ ID No: 25. LCDR3 and V having the sequence as in SEQ ID No: 30 H CDR3.

[0039] The present invention provides an antibody that binds to human CD38, comprising a human light chain and a human heavy chain variable region, wherein the light chain variable region comprises a V having a sequence as shown in SEQ ID No: 23. L CDR1, V having the sequence in SEQ ID No: 24 L CDR2 and V having the sequence in SEQ ID No: 25 L CDR3, the heavy chain variable region comprises a V having a sequence as in SEQ ID No: 28 H CDR1, V having the sequence in SEQ ID No: 29 H CDR2 and V having the sequence in SEQ ID No: 30 H CDR3.

[0040] The present invention provides an antibody that binds to human CD38, comprising a V having an amino acid sequence as shown in SEQ ID No: 22. L district.

[0041] The present invention provides an antibody that binds to human CD38, comprising a V sequence having at least about 90%, such as at least about 95%, amino acid sequence identity to the sequence in SEQ ID No: 22. L district.

[0042] The present invention provides an antibody that binds to human CD38, comprising a V having an amino acid sequence as shown in SEQ ID No: 27. H district.

[0043] The present invention provides an antibody that binds to human CD38, comprising a V sequence spanning SEQ ID No: 27. H CDR1-V H The amino acid sequence of the CDR3 region is V H district.

[0044] The present invention provides an antibody that binds to human CD38, which comprises a sequence as in SEQ ID No: 27 or a sequence spanning V as in SEQ ID No: 27. H CDR1-V H The region of CDR3 has at least about 90%, such as at least about 95%, amino acid sequence identity to V H district.

[0045] The present invention provides an antibody that binds to human CD38, which comprises a sequence as in SEQ ID No: 27 or a sequence spanning V as in SEQ ID No: 27. H CDR1-V H The CDR3 region has 1-5, for example 1-3 amino acid substitutions, deletions or additions. H district.

[0046] The present invention provides an antibody that binds to human CD38, which comprises V as defined above. L region and V as defined above H district.

[0047] The present invention provides a peptide that binds to human CD38 (SEQ ID No: 31), and the extent of its binding to mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is substituted with a phenylalanine residue is different from the extent of its binding to human CD38 (SEQ ID No: 31).

[0048] The present invention provides a peptide as defined above, wherein the peptide binds to an EC of a mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is substituted with a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 50%, such as less than 10%, less than 5% or less than 1%.

[0049] The present invention provides a peptide that binds to human CD38 (SEQ ID No: 31), and the extent of its binding to mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is substituted with an arginine residue is different from the extent of its binding to human CD38 (SEQ ID No: 31).

[0050] The present invention provides a peptide as defined above, wherein the peptide binds to an EC of a mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 50%, such as less than 10%, less than 5% or less than 1%.

[0051] The present invention provides a peptide as defined above, wherein the peptide binds to a mutant human CD38 (SEQ ID No: 32) in which the threonine residue at position 237 is replaced by an alanine residue to the same extent as it binds to human CD38 (SEQ ID No: 31).

[0052] The present invention provides a peptide as defined above, wherein the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 EC equivalent to peptide binding to human CD38 (SEQ ID No: 31) 50 75%-125% of.

[0053] The present invention provides a peptide that binds to human CD38 (SEQ ID No: 31), wherein the peptide has the following binding properties: (i) binds to a mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is substituted with a phenylalanine residue to the same extent as its binding to human CD38 (SEQ ID No: 31), (ii) binds to a mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is substituted with an arginine residue to the same extent as its binding to human CD38 (SEQ ID No: 31), and (iii) binds to a mutant human CD38 (SEQ ID No: 32) in which the threonine residue at position 237 is substituted with an alanine residue to the same extent as its binding to human CD38 (SEQ ID No: 31).

[0054] The present invention provides a peptide that binds to human CD38 (SEQ ID No: 31), wherein the peptide has the following binding properties: (i) it binds to a mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is substituted by a phenylalanine residue to a degree different from its binding to human CD38 (SEQ ID No: 31), (ii) it binds to a mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is substituted by an arginine residue to a degree different from its binding to human CD38 (SEQ ID No: 31), and (iii) it binds to a mutant human CD38 (SEQ ID No: 32) in which the threonine residue at position 237 is substituted by an alanine residue to the same degree as its binding to human CD38 (SEQ ID No: 31).

[0055] The present invention provides a peptide as defined above, wherein EC 50 The assay was performed by ELISA as described in Example 17 of the specification.

[0056] The present invention provides peptides that compete for binding to CD38 with the antibody described in technical solution (i) above. In one technical solution, the competition is determined by the ELISA described in Example 8 or 9 of the specification, wherein competition is defined as at least 90% of the signal measured by absorbance, or by the cross-blocking assay described in Example 7 of the specification, wherein competition is defined as at least 90% of the signal measured by fluorescence.

[0057] The present invention provides peptides that specifically bind to a CD38 antigenic determinant, which antigenic determinant also specifically binds to an antibody as defined above.

[0058] The present invention provides a peptide that specifically binds to the SKRNIQFSCKNIYR region and the EKVQTLEAWVIHGG region of human CD38 (SEQ ID No: 31).

[0059] The present invention provides peptides having substantially the same specific binding properties as the antibodies defined above with respect to binding to human CD38.

[0060] The present invention provides peptides that bind to human CD38, wherein the antibodies have one or more of the following characteristics:

[0061] (i) as an antagonist of CD38;

[0062] (ii) does not induce significant proliferation of peripheral blood mononuclear cells when measured by the method described in Example 18 of the specification;

[0063] (iii) does not induce significant release of IL-6 from human monocytes or peripheral blood mononuclear cells when measured by the method described in Example 19 of the specification;

[0064] (iv) does not induce detectable IFN-γ release from human T cells or peripheral blood mononuclear cells when measured by the method described in Example 20 of the specification;

[0065] (v) internalized by CD38-expressing cells; for example, CHO-CD38 can be internalized by maintaining at 37° C. for 5 to 15 minutes by the method described in Example 12 of the specification;

[0066] (vi) Induction of ADCC; for example, EC in Daudi-luc cells 50 Values ​​below 15 ng / ml; for example, below 10 ng / ml, such as in MM cells EC 50 The value is less than 75 ng / ml, less than 50 ng / ml, 30 ng / ml or 10 ng / ml, as determined by the method described in Example 5 of the specification;

[0067] (vii) Induction of CDC in the presence of complement; for example, EC in daudi-luc or CD38-CHO cells 50 The value is less than 5 μg / ml, for example less than 1 μg / ml, as determined by the method described in Example 6 of the specification;

[0068] (viii) inhibiting the synthesis of cGDPR;

[0069] (ix) inhibiting the synthesis of cADPR;

[0070] (x) Binding affinity to human CD38 (K D ) is less than 10 -8 M, for example, the range is 10 -8 M to 10 -11 M, 7×10 -9 M to 10 -10 M, measured by surface plasmon resonance as described in Example 20 of the specification.

[0071] The present invention provides a peptide as defined above, which can inhibit the synthesis of cGDPR by at least 25%, such as at least 30%, at a concentration of 3 μg / ml in 90 minutes, as determined photometrically as described in Example 24 of the specification.

[0072] The present invention provides a peptide as defined above, which can inhibit the synthesis of cADPR by at least 25%, such as at least 30%, at a concentration of 3 μg / ml in 90 minutes, as determined by the HPLC method described in Munshi et al., J. Biol. Chem. 275, 21566-21571 (2000).

[0073] In one embodiment, the peptide as defined above is a human monoclonal antibody.

[0074] The present invention provides an antibody as defined above, characterized in that it is a full-length IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE or IgM antibody, such as an IgG1 antibody, preferably an IgG1, kappa antibody, or an IgM antibody, preferably an IgM, kappa antibody.

[0075] The present invention provides isolated human monoclonal antibodies comprising

[0076] (i) Derived from human Hv1263 / 3M28 (V H I) a heavy chain variable region amino acid sequence that is a germline sequence and a light chain variable region amino acid sequence that is derived from a human L15 (VκI) germline sequence, wherein the human antibody binds to human CD38; or

[0077] (ii) Derived from human V H 3-DP-47 / V3-23(V H III) a heavy chain variable region amino acid sequence of a germline sequence and a light chain variable region amino acid sequence derived from a human L6 (VκI) germline sequence, wherein the human antibody binds to human CD38.

[0078] The present invention provides a peptide as defined above, wherein the peptide is glycosylated in a eukaryotic cell.

[0079] In one embodiment, the antibody of the present invention is an antibody fragment or a single-chain antibody.

[0080] The present invention provides a peptide as defined above, further comprising a chelator linking region for attaching a radioisotope.

[0081] The present invention provides a peptide as defined above, in substantially isolated form.

[0082] The present invention provides an isolated nucleic acid encoding a peptide as defined above.

[0083] The present invention provides an expression vector comprising a nucleic acid sequence encoding a peptide as defined above.

[0084] The present invention provides an expression vector comprising

[0085] (i) V in SEQ ID No: 1 L Nucleotide sequence;

[0086] (ii) V in SEQ ID No: 6 H Nucleotide sequence;

[0087] (iii) V in SEQ ID No: 1 L Nucleotide sequence and V in SEQ ID No: 6 H Nucleotide sequence;

[0088] (iv) V in SEQ ID No: 11 L Nucleotide sequence;

[0089] (v) V in SEQ ID No: 16 H Nucleotide sequence;

[0090] (vi) V in SEQ ID No: 11 L Nucleotide sequence and V in SEQ ID No: 16 H Nucleotide sequence;

[0091] (vii) V in SEQ ID No: 21 L Nucleotide sequence;

[0092] (viii) V in SEQ ID No: 26 H nucleotide sequence; or

[0093] (ix) V in SEQ ID No: 21 L Nucleotide sequence and V in SEQ ID No: 26 H Nucleotide sequence.

[0094] The present invention provides an expression vector as defined above, further comprising a nucleotide sequence encoding the constant region of the light chain, heavy chain, or light and heavy chains of a human antibody.

[0095] The present invention provides an expression vector as defined above, wherein the nucleotide sequence encoding the constant region of the light chain, heavy chain, or light and heavy chains of a human antibody encodes an IgG1 antibody.

[0096] The present invention provides a hybridoma that produces a human monoclonal anti-CD38 antibody encoded by human light chain and human heavy chain nucleic acids, comprising

[0097] (i) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 1 and SEQ ID No: 6, respectively, in their variable regions;

[0098] (ii) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 11 and SEQ ID No: 16, respectively, in their variable regions;

[0099] (iii) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 21 and SEQ ID No: 26, respectively, in their variable regions; or

[0100] (iv) Human light chain and human heavy chain nucleic acids containing in their variable regions the conservative sequence changes listed in (i), (ii) or (iii).

[0101] The present invention provides hybridomas that produce human monoclonal anti-CD38 antibodies having human heavy and light chain variable regions comprising the following sequences:

[0102] (i) the human light chain variable amino acid sequence in SEQ ID No: 2 and the human heavy chain variable amino acid sequence in SEQ ID No: 7;

[0103] (ii) a human light chain variable amino acid sequence as in SEQ ID No: 12 and a human heavy chain variable amino acid sequence as in SEQ ID No: 17;

[0104] (iii) a human light chain variable amino acid sequence as in SEQ ID No: 22 and a human heavy chain variable amino acid sequence as in SEQ ID No: 27; or

[0105] (iv) human light chain and human heavy chain variable amino acid sequences with conservative sequence variations listed in (i), (ii) or (iii).

[0106] The present invention provides a transfectoma that produces a human monoclonal anti-CD38 antibody encoded by a human light chain and a human heavy chain nucleic acid comprising the following sequences:

[0107] (i) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 1 and SEQ ID No: 6, respectively, in their variable regions;

[0108] (ii) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 11 and SEQ ID No: 16, respectively, in their variable regions;

[0109] (iii) human light chain and human heavy chain nucleic acids comprising the nucleotide sequences shown in SEQ ID No: 21 and SEQ ID No: 26, respectively, in their variable regions; or

[0110] (iv) Human light chain and human heavy chain nucleic acids containing in their variable regions the conservative sequence changes listed in (i), (ii) or (iii).

[0111] The present invention provides a transfectoma that produces a human monoclonal anti-CD38 antibody having human heavy and light chain variable regions comprising the following sequences:

[0112] (i) the human light chain variable amino acid sequence in SEQ ID No: 2 and the human heavy chain variable amino acid sequence in SEQ ID No: 7;

[0113] (ii) a human light chain variable amino acid sequence as in SEQ ID No: 12 and a human heavy chain variable amino acid sequence as in SEQ ID No: 17;

[0114] (iii) a human light chain variable amino acid sequence as in SEQ ID No: 22 and a human heavy chain variable amino acid sequence as in SEQ ID No: 27; or

[0115] (iv) human light chain and human heavy chain variable amino acid sequences with conservative sequence variations listed in (i), (ii) or (iii).

[0116] The present invention provides eukaryotic or prokaryotic host cells capable of producing the peptides defined above.

[0117] The present invention provides a eukaryotic or prokaryotic host cell containing an expression vector as defined above.

[0118] The present invention provides a transgenic non-human animal or plant comprising a nucleic acid encoding a human heavy chain and a human light chain, wherein the animal or plant produces a detectable amount of a peptide as defined above.

[0119] The present invention provides an immunoconjugate comprising a peptide as defined above linked to a cytotoxic agent, a radioisotope or a drug.

[0120] The present invention provides an immunoconjugate comprising a peptide as defined above, wherein the peptide is a monovalent IgM antibody linked to a cytotoxic agent, a radioisotope or a drug.

[0121] The present invention provides bispecific or multispecific molecules comprising a peptide as defined above and having binding specificity for human effector cells.

[0122] The present invention provides bispecific or multispecific molecules comprising a peptide as defined above and having binding specificity for CD3, CD4, CD138, IL-15R, membrane-bound or receptor-bound TNF-α, human Fc receptor, or membrane-bound or receptor-bound IL-15.

[0123] The present invention provides a pharmaceutical composition comprising the peptide as defined above or the immunoconjugate as defined above and a pharmaceutically acceptable carrier.

[0124] The present invention provides a pharmaceutical composition as defined above containing one or more additional therapeutic agents.

[0125] The present invention provides a method for inhibiting the growth and / or proliferation of cells expressing CD38, comprising administering the peptide defined above, the immunoconjugate defined above, the pharmaceutical composition defined above, or the expression vector defined above, thereby inhibiting the growth and / or proliferation of cells.

[0126] The present invention provides a method for treating a disease or condition in a patient in which cells expressing CD38 are involved, the method comprising administering to a patient in need of treatment a peptide as defined above, an immunoconjugate as defined above, a pharmaceutical composition as defined above, or an expression vector as defined above.

[0127] The present invention provides a method for inhibiting a disease or symptom in a patient in which cells expressing CD38 participate, which comprises administering to a patient in need of treatment a peptide as defined above, an immunoconjugate as defined above, a pharmaceutical composition as defined above, or an expression vector as defined above.

[0128] In one embodiment, the disease or condition is rheumatoid arthritis.

[0129] In one embodiment, the disease or condition is multiple myeloma.

[0130] In one embodiment, the method includes administering to the patient one or more additional therapeutic agents.

[0131] In one embodiment, the one or more other therapeutic agents are selected from chemotherapeutic agents, anti-inflammatory agents, or immunosuppressive and / or immunomodulatory agents.

[0132] In one embodiment, the one or more other therapeutic agents are selected from platinum, gefitinib, cetuximab, rituximab, bevacizumab, erlotinib, bortezomib, thalidomide, pamidronate, zoledronic acid, clodronate, risedronate, ibandronate, etidronate, alendronate, tiludronate, arsenic trioxide, lenalidomide, filgrastim, pegfilgrastim, sargramostim, N-suberoylanilide hydroxamic acid and SCIO-469.

[0133] The present invention provides an in vitro method for detecting the presence of CD38 antigen or cells expressing CD38 in a sample, comprising: a) contacting the sample with a peptide as defined above under conditions such that the peptide and CD38 can form a complex; and b) detecting the formation of the complex.

[0134] The present invention provides a kit for detecting the presence of CD38 antigen or cells expressing CD38 in a sample containing the peptide defined above.

[0135] The present invention provides an in vivo method for detecting CD38 antigen or cells expressing CD38 in a patient, comprising:

[0136] a) administering a peptide as claimed in any one of claims 1 to 55 under conditions in which the peptide and CD38 can form a complex; and

[0137] b) Detecting the formed complex.

[0138] The present invention provides anti-idiotypic antibodies that bind to an antibody as defined above.

[0139] In one embodiment, an anti-idiotypic antibody is used to detect the level of an antibody as defined above in a sample.

[0140] In one embodiment, an anti-idiotypic antibody is used to detect the level of human monoclonal antibodies against CD38 in a sample. BRIEF DESCRIPTION OF THE DRAWINGS

[0141] Figure 1Figure A shows the binding of -003, -005 and isotype control antibody HuMab-KLH to CD38 transfected CHO (CHO-CD38) cells as determined by flow cytometry. The experimental setup is described in Example 4.

[0142] Figure 1 B shows the binding of HuMab-024 and HuMab-KLH to CD38-transfected CHO (CHO-CD38) cells as determined by flow cytometry. The experimental setup is described in Example 4.

[0143] Figure 2 Figure A shows the binding of -003, -005 and isotype control antibody HuMab-KLH to Daudi cells as determined by flow cytometry. The experimental setup is described in Example 4.

[0144] Figure 2 B shows the binding of -024 and HuMab-KLH to Daudi cells as determined by flow cytometry. The experimental setup is described in Example 4.

[0145] Figure 3 Shown is the binding of -003, -005, -024 and HuMab-KLH to multiple myeloma cells. The experimental setup is described in Example 4.

[0146] Figure 4 A shows the ability of -003 and -005 to induce Daudi cell lysis by ADCC compared to rituximab and HuMab-KLH. The experimental setup is described in Example 5.

[0147] Figure 4 Shown in B is the ability of -024 to induce Daudi cell lysis by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

[0148] Figure 5 A shows the ability of -003, -005 and -024 to induce lysis of fresh multiple myeloma cells by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

[0149] Figure 5 Shown in B is the ability of -003, -005, and -024 to induce lysis of fresh plasma cell leukemia tumor cells by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

[0150] Figure 6Shown is the ability of -003 and -005 to induce lysis of JK6L (a multiple myeloma cell line) by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

[0151] Figure 7 Shown is the ability of -003 and -005 to induce lysis of AMO-1 (a multiple myeloma cell line) by ADCC compared to HuMab-KLH. The experimental setup is described in Example 5.

[0152] Figure 8 Shown is the CDC-mediated lysis of Daudi-luc cells induced by -003 and -005 compared to HuMab-KLH. The experimental setup is described in Example 6.

[0153] Figure 9 A shows CDC-mediated lysis of CHO-CD38 cells induced by -003 and -005 compared to HuMab-KLH. The experimental setup is described in Example 6.

[0154] Figure 9 B shows CDC-mediated lysis of CHO-CD38 cells induced by -024 compared to HuMab-KLH. The experimental setup is described in Example 6.

[0155] Figure 10 A shows CDC-mediated lysis of 3% resistant tumor cells in the presence of -003, -005 and HuMab-KLH. The experimental setup is described in Example 6.

[0156] Figure 10 B shows CDC-mediated lysis of 9% resistant tumor cells in the presence of -003, -005 and HuMab-KLH. The experimental setup is described in Example 6.

[0157] Figure 10 C shows CDC-mediated lysis of 30-40% resistant tumor cells in the presence of -003, -005 and HuMab-KLH. The experimental setup is described in Example 6.

[0158] Figure 10 D shows CDC-mediated lysis of 70% resistant tumor cells in the presence of -003, -005 and HuMab-KLH. The experimental setup is described in Example 6.

[0159] Figure 10 E shows CDC-mediated lysis of multiple myeloma cells in the presence of -024 and HuMab-KLH. The experimental setup is described in Example 6.

[0160] Figure 11 It is shown that -003 and -005 do not cross-inhibit binding to CD38. The experimental setup is described in Example 7.

[0161] Figure 12A Shown are immunohistochemical staining of macrophages, lymphocytes and plasma B cells containing -003. The experimental setup is described in Example 10.

[0162] Figure 12B Shown is immunohistochemical staining of bronchial epithelial cells containing -003. The experimental setup is described in Example 10.

[0163] Figure 12C Shown is immunohistochemical staining of myocytes containing -003. The experimental setup is described in Example 10.

[0164] Figure 12D Shown is immunohistochemical staining of macaque lymphoid tissue containing -003. The experimental setup is described in Example 10.

[0165] Figure 13A Shown are immunohistochemical staining of macrophages, lymphocytes, and plasma B cells containing -005. The experimental setup is described in Example 10.

[0166] Figure 13B Shown is immunohistochemical staining of bronchial epithelial cells containing -005. The experimental setup is described in Example 10.

[0167] Figure 13C Shown is immunohistochemical staining of myocytes containing -005. The experimental setup is described in Example 10.

[0168] Figure 13D Shown is immunohistochemical staining of macaque lymphoid tissue containing -005. The experimental setup is described in Example 10.

[0169] Figure 14 A shows immunohistochemical staining of liver endothelial cells containing CD31. The experimental setup is described in Example 10.

[0170] Figure 14 B shows immunohistochemical staining of liver endothelial cells containing vWF. The experimental setup is described in Example 10.

[0171] Figure 14 C shows immunohistochemical staining of liver endothelial cells with anti-KLH. The experimental setup is described in Example 10.

[0172] Figure 14D shows immunohistochemical staining of liver endothelial cells containing -003. The experimental setup is described in Example 10.

[0173] Figure 14 E shows immunohistochemical staining of liver endothelial cells in the presence of -005. The experimental setup is described in Example 10.

[0174] Figure 15A Shown is the cross-reactivity of -003 and -005 compared to HuMab-KLH on macaque lymphocytes as determined by flow cytometry. The experimental setup is described in Example 11.

[0175] Figure 15B Shown is the cross-reactivity of -003 and -005 compared to HuMab-KLH on macaque monocytes as determined by flow cytometry. The experimental setup is described in Example 11.

[0176] Figure 15C Shown is the cross-reactivity of -003 and -005 compared to HuMab-KLH on rhesus monkey PBMCs as determined by flow cytometry. The experimental setup is described in Example 11.

[0177] Figure 16A Shown is the internalization of -003 as determined by EtBr quenching. The experimental setup is described in Example 12.

[0178] Figure 16B Shown is the internalization of -005 as determined by EtBr quenching. The experimental setup is described in Example 12.

[0179] Figure 17A Shown are the inhibitory effects of -003 and -005 on tumor cell growth in a prophylactic setting compared to anti-CD20 monoclonal antibody (rituximab) and HuMab-KLH as measured by in vivo SCID fluorescence imaging. The experimental setup is described in Example 13.

[0180] Figure 17B Shown are the inhibitory effects of -003 and -005 on tumor cell growth compared to anti-CD20 monoclonal antibody (rituximab) and HuMab-KLH in therapeutic setting I as measured by in vivo SCID fluorescence imaging. The experimental setup is described in Example 13.

[0181] Figure 17C Shown are the inhibitory effects of -003 and -005 on tumor cell growth in therapeutic setting II compared to anti-CD20 monoclonal antibody (rituximab) and HuMab-KLH as measured by in vivo SCID fluorescence imaging. The experimental setup is described in Example 13.

[0182] Figure 17D Shown are the inhibitory effects of -003 and -024 on tumor cell growth compared to HuMab-KLH in therapeutic setting III as measured by in vivo SCID fluorescence imaging. The experimental setup is described in Example 13.

[0183] Figure 18 Shown are the induction of apoptosis by -003 and -024 when uncrosslinked or crosslinked compared to anti-CD20 monoclonal antibody (rituximab) and HuMab-KLH. The experimental setup is described in Example 14.

[0184] Figure 19 Shown are the histological scores of CD38-positive cells in xenografts transplanted into RA-SCID mice 14 days after treatment with anti-KLH (HuMab-KLH) or -005. The methods are described in Example 15.

[0185] Figure 20 Shown are histological scores of CD38-positive cells in xenografts transplanted into RA-SCID mice 14 days after treatment with anti-KLH or -005. Methods are described in Example 15.

[0186] Figure 21 Shown are CD38 staining of B cells from xenografts before transplantation (A) or after treatment with anti-KLH (B) or -005 (C). Methods are described in Example 15.

[0187] Figure 22 Shown are CD138 staining of B cells from xenografts before transplantation (A) or after treatment with anti-KLH (B) or -005 (C). Methods are described in Example 15.

[0188] Figure 23 Shown is the binding of -003 and -005 to wild-type and mutant human CD38 as determined by ELISA.

[0189] 23A: Binding of -003 and -005 to T237A mutant human CD38.

[0190] 23B: Binding of -003 and -005 to Q272R mutant human CD38.

[0191] 23C: Binding of -003 and -005 to S274F mutant human CD38. Methods are described in Example 17.

[0192] Figure 24Shown are the effects of -003 and -005 on proliferation (A), IL-6 production (B), and IFN-γ production (C) of human PBMCs compared to HuMab-KLH. Methods are as described in Examples 18, 19, and 20, respectively.

[0193] Figure 25 Shown is the enzymatic production of cGDP ribose in the presence of various concentrations of -003 (B), -005 (C), -024 (D) or anti-KLH (A). The method is described in Example 23.

[0194] Figure 26 Shown is a comparison of -003, -005, and Morphosys antibody TH-3079 in CDC of CHO-CD38 cells (26A), CDC of Daudi cells (26B), and ADCC of Daudi cells (26C).

[0195] The sequences of the present invention are shown in the attached sequence listing.

[0196] SEQ ID No: 1 is antibody-003V L The nucleotide sequence of the region.

[0197] SEQ ID No: 2 is antibody-003V L The amino acid sequence of the region.

[0198] SEQ ID No: 3 is antibody-003V L The amino acid sequence of CDR1 includes amino acids 24-34 of SEQ ID No: 2.

[0199] SEQ ID No: 4 is antibody-003V L The amino acid sequence of CDR2 includes amino acids 50-56 of SEQ ID No: 2.

[0200] SEQ ID No: 5 is antibody-003V L The amino acid sequence of CDR3 includes amino acids 89-97 of SEQ ID No: 2.

[0201] SEQ ID No: 6 is antibody-003V H The nucleotide sequence of the region.

[0202] SEQ ID No: 7 is antibody-003V H The amino acid sequence of the region.

[0203] SEQ ID No: 8 is antibody-003V HThe amino acid sequence of CDR1 includes amino acids 31-35 of SEQ ID No: 7.

[0204] SEQ ID No: 9 is antibody-003V H The amino acid sequence of CDR2 includes amino acids 50-66 of SEQ ID No: 7.

[0205] SEQ ID No: 10 is antibody-003V H The amino acid sequence of CDR3 includes amino acids 99-109 of SEQ ID No: 7.

[0206] SEQ ID No: 11 is antibody-005V L The nucleotide sequence of the region.

[0207] SEQ ID No: 12 is antibody-005V L The amino acid sequence of the region.

[0208] SEQ ID No: 13 is antibody-005V L The amino acid sequence of CDR1 includes amino acids 24-34 of SEQ ID No: 12.

[0209] SEQ ID No: 14 is antibody-005V L The amino acid sequence of CDR2 includes amino acids 50-56 of SEQ ID No: 12.

[0210] SEQ ID No: 15 is antibody-005V L The amino acid sequence of CDR3 includes amino acids 89-97 of SEQ ID No: 12.

[0211] SEQ ID No: 16 is antibody-005V H The nucleotide sequence of the region.

[0212] SEQ ID No: 17 is antibody-005V H The amino acid sequence of the region.

[0213] SEQ ID No: 18 is antibody-005V H The amino acid sequence of CDR1 includes amino acids 31-35 of SEQ ID No: 17.

[0214] SEQ ID No: 19 is antibody-005V H The amino acid sequence of CDR2 includes amino acids 50-66 of SEQ ID No: 17.

[0215] SEQ ID No: 20 is antibody-005V H The amino acid sequence of CDR3 includes amino acids 99-111 of SEQ ID No: 17.

[0216] SEQ ID No: 21 is antibody-024V L The nucleotide sequence of the region.

[0217] SEQ ID No: 22 is antibody-024V L The amino acid sequence of the region.

[0218] SEQ ID No: 23 is antibody-024V L The amino acid sequence of CDR1 includes amino acids 24-34 of SEQ ID No: 22.

[0219] SEQ ID No: 24 is antibody-024V L The amino acid sequence of CDR2 includes amino acids 50-66 of SEQ ID No: 22.

[0220] SEQ ID No: 25 is antibody-024V L The amino acid sequence of CDR3 includes amino acids 89-97 of SEQ ID No: 22.

[0221] SEQ ID No: 26 is antibody-024V H The nucleotide sequence of the region.

[0222] SEQ ID No: 27 is antibody-024V H The amino acid sequence of the region.

[0223] SEQ ID No: 28 is antibody-024V H The amino acid sequence of CDR1 includes amino acids 31-35 of SEQ ID No: 27.

[0224] SEQ ID No: 29 is antibody-024V H The amino acid sequence of CDR2 includes amino acids 50-66 of SEQ ID No: 27.

[0225] SEQ ID No: 30 is antibody-024V H The amino acid sequence of CDR3 includes amino acids 99-111 of SEQ ID No: 27.

[0226] SEQ ID No: 31 is the sequence of human CD38.

[0227] SEQ ID No: 32 is a mutant human CD38 sequence, wherein the threonine at position 237 is replaced by alanine.

[0228] SEQ ID No: 33 is a mutant human CD38 sequence, wherein glutamic acid at position 272 is replaced by arginine.

[0229] SEQ ID No: 34 is a mutant human CD38 sequence, wherein the serine at position 274 is replaced by phenylalanine. Specific implementation plan

[0230] The present invention provides peptides that bind to CD38 ("CD38BPs"), which are useful for treating, diagnosing, and preventing various diseases in which cells expressing CD38 are involved, such as multiple myeloma.

[0231] In one embodiment, the CD38BP of the present invention is antibody -003. -003 is a human monoclonal IgG1 antibody having a V sequence including SEQ ID No: 2. L Region and V comprising the sequence of SEQ ID No: 7 H district.

[0232] In one embodiment, the CD38BP of the present invention is antibody -005. -005 is a human monoclonal IgG1 antibody having a V sequence including SEQ ID No: 12. L region and a V region comprising the sequence of SEQ ID No: 17 H district.

[0233] In one embodiment, the CD38BP of the present invention is antibody -024. -024 is a human monoclonal IgG1 antibody having a V sequence including SEQ ID No: 22. L region and a V region comprising the sequence of SEQ ID No: 27 H district.

[0234] Antibodies interact with target antigens primarily through the amino acid residues located in the six heavy and light chain complementary determining regions (CDRs). Therefore, between individual antibodies, the amino acid sequences in the CDRs vary more than the sequences outside the CDRs. Since the CDR sequences are responsible for most antibody-antigen interactions, recombinant antibodies that can mimic the properties of specific naturally occurring antibodies can be expressed by transplanting the CDR sequences from specific naturally occurring antibodies into expression vectors constructed from the same frame sequences of different antibodies with different properties (see, for example, Riechmann, L. et al., Nature 332, 323-327 (1998), Jones, P. et al., Nature 321, 522-525 (1986) and Queen, C. et al., PNAS USA 86, 10029-10033 (1989)).

[0235] Since it is known in the art that the antibody heavy chain CDR3 domain plays a very important role in the binding specificity / affinity of the antibody to the antigen (Ditzel HJ, et al., J Immunol. 157(2), 739-49 (1996), Barbas SM et al., J. Am. Chem. Soc. 116, 2161-2162 (1994), and Barbas SM et al., Proc Natl Acad Sci USA 92(7), 2529-33 (1995)), the CD38BPs of the present invention may include the heavy chain CDR3s of -003, -005, or -024. The CD38BPs of the present invention also include the light chain CDR3s of -003, -005, or -024. The CD38BPs of the present invention further include the heavy chain CDR2s of -003, -005, or -024, respectively. The CD38BPs of the present invention further include the heavy chain CDR1s of -003, -005, or -024, respectively.

[0236] The present invention provides CD38BPs that can compete with -003 for binding to CD38.

[0237] The present invention provides CD38BPs that can compete with -005 for binding to CD38.

[0238] The present invention provides CD38BPs that can compete with -024 for binding to CD38.

[0239] In one embodiment, competition is determined by ELISA as described in the Examples section.

[0240] In one embodiment, competition is determined by FACS as described in the Examples section.

[0241] The present invention provides a CD38BP that binds to a CD38 antigenic determinant that also specifically binds to -003, -005, or -024.

[0242] The present invention provides a CD38BP having substantially the same specific binding properties for human CD38 as -003, -005, or -024.

[0243] The present invention provides a V comprising mainly consisting of SEQ ID No: 3 L CD38BP of CDR1.

[0244] The present invention provides a V comprising mainly consisting of SEQ ID No: 4 L CD38BP of CDR2.

[0245] The present invention provides a V comprising mainly consisting of SEQ ID No: 5 L CD38BP of CDR3.

[0246] The present invention provides a V comprising mainly consisting of SEQ ID No: 5 L CDR3 and V consisting mainly of SEQ ID No: 3 L CD38BP of CDR1.

[0247] The present invention provides a V comprising mainly consisting of SEQ ID No: 5 L CDR3 and V consisting mainly of SEQ ID No: 4 L CD38BP of CDR2.

[0248] The present invention provides a V comprising mainly consisting of SEQ ID No: 5 L CDR3 and V consisting mainly of SEQ ID No: 4 L CDR2 and V consisting mainly of SEQ ID No: 3 L CD38BP of CDR1.

[0249] The present invention provides a V comprising mainly consisting of SEQ ID No: 8 H CD38BP of CDR1.

[0250] The present invention provides a V comprising mainly consisting of SEQ ID No: 9 H CD38BP of CDR2.

[0251] The present invention provides a V comprising mainly consisting of SEQ ID No: 10 H CD38BP of CDR3.

[0252] The present invention provides a V comprising mainly consisting of SEQ ID No: 10 H CDR3 and V consisting mainly of SEQ ID No: 8 H CD38BP of CDR1.

[0253] The present invention provides a V comprising mainly consisting of SEQ ID No: 10 H CDR3 and V consisting mainly of SEQ ID No: 9 H CD38BP of CDR2.

[0254] The present invention provides a V comprising mainly consisting of SEQ ID No: 10 H CDR3 and V consisting mainly of SEQ ID No: 9 H CDR2 and V consisting mainly of SEQ ID No: 8 H CD38BP of CDR1.

[0255] The present invention provides a V comprising mainly consisting of SEQ ID No: 13 L CD38BP of CDR1.

[0256] The present invention provides a V comprising mainly consisting of SEQ ID No: 14 L CD38BP of CDR2.

[0257] The present invention provides a V comprising mainly consisting of SEQ ID No: 15 L CD38BP of CDR3.

[0258] The present invention provides a V comprising mainly consisting of SEQ ID No: 15 L CDR3 and V consisting mainly of SEQ ID No: 13 L CD38BP of CDR1.

[0259] The present invention provides a V comprising mainly consisting of SEQ ID No: 15 L CDR3 and V consisting mainly of SEQ ID No: 14 L CD38BP of CDR2.

[0260] The present invention provides a V comprising mainly consisting of SEQ ID No: 15 L CDR3 and V consisting mainly of SEQ ID No: 14 L CDR2 and V consisting mainly of SEQ ID No: 13 L CD38BP of CDR1.

[0261] The present invention provides a V comprising mainly consisting of SEQ ID No: 18 H CD38BP of CDR1.

[0262] The present invention provides a V comprising mainly consisting of SEQ ID No: 19 H CD38BP of CDR2.

[0263] The present invention provides a V comprising mainly consisting of SEQ ID No: 20 H CD38BP of CDR3.

[0264] The present invention provides a V comprising mainly consisting of SEQ ID No: 20 H CDR3 and V consisting mainly of SEQ ID No: 18 H CD38BP of CDR1.

[0265] The present invention provides a V comprising mainly consisting of SEQ ID No: 20 H CDR3 and V consisting mainly of SEQ ID No: 19 H CD38BP of CDR2.

[0266] The present invention provides a V comprising mainly consisting of SEQ ID No: 20 H CDR3 and V consisting mainly of SEQ ID No: 19 H CDR2 and V consisting mainly of SEQ ID No: 18 H CD38BP of CDR1.

[0267] The present invention provides a V comprising mainly consisting of SEQ ID No: 23 L CD38BP of CDR1.

[0268] The present invention provides a V comprising mainly consisting of SEQ ID No: 24 L CD38BP of CDR2.

[0269] The present invention provides a V comprising mainly consisting of SEQ ID No: 25 L CD38BP of CDR3.

[0270] The present invention provides a V comprising mainly consisting of SEQ ID No: 25 L CDR3 and V consisting mainly of SEQ ID No: 23 L CD38BP of CDR1.

[0271] The present invention provides a V comprising mainly consisting of SEQ ID No: 25 LCDR3 and V consisting mainly of SEQ ID No: 24 L CD38BP of CDR2.

[0272] The present invention provides a V comprising mainly consisting of SEQ ID No: 25 L CDR3 and V consisting mainly of SEQ ID No: 24 L CDR2 and V consisting mainly of SEQ ID No: 23 L CD38BP of CDR1.

[0273] The present invention provides a V comprising mainly consisting of SEQ ID No: 28 H CD38BP of CDR1.

[0274] The present invention provides a V comprising mainly consisting of SEQ ID No: 29 H CD38BP of CDR2.

[0275] The present invention provides a V comprising mainly consisting of SEQ ID No: 30 H CD38BP of CDR3.

[0276] The present invention provides a V comprising mainly consisting of SEQ ID No: 30 H CDR3 and V consisting mainly of SEQ ID No: 28 H CD38BP of CDR1.

[0277] The present invention provides a V comprising mainly consisting of SEQ ID No: 30 H CDR3 and V consisting mainly of SEQ ID No: 29 H CD38BP of CDR2.

[0278] The present invention provides a V comprising mainly consisting of SEQ ID No: 30 H CDR3 and V consisting mainly of SEQ ID No: 29 H CDR2 and V consisting mainly of SEQ ID No: 28 H CD38BP of CDR1.

[0279] The present invention provides CD38BP, comprising

[0280] (a) Contains 3 V L The first V of CDRs L regions, each of which consists essentially of SEQ ID No: 3, SEQ ID No: 4, and SEQ ID No: 5; and

[0281] (b) Contains 3 V H CDRS's first V H regions, each of which consists essentially of SEQ ID No: 8, SEQ ID No: 9, and SEQ ID No: 10.

[0282] The present invention provides CD38BP, comprising

[0283] (a) Contains 3 V L The first V of CDRs L regions, each of which consists essentially of SEQ ID No: 13, SEQ ID No: 14, and SEQ ID No: 15; and

[0284] (b) Contains 3 V H The first V of CDRs H regions, each of which consists essentially of SEQ ID No: 18, SEQ ID No: 19, and SEQ ID No: 20.

[0285] The present invention provides CD38BP, comprising

[0286] (a) Contains 3 V L The first V of CDRs L regions, each of which consists essentially of SEQ ID No: 23, SEQ ID No: 24, and SEQ ID No: 25; and

[0287] (b) Contains 3 V H CDRS's first V H regions, each of which consists essentially of SEQ ID No: 28, SEQ ID No: 29, and SEQ ID No: 30.

[0288] In one technical solution, V L Area and V H The regions are located on the same chain of the peptide.

[0289] In another technical solution, V L Area and V H The regions are separated by flexible connectors.

[0290] In one technical solution, V L Area and V H The regions are located on separate chains of the peptide.

[0291] In another technical solution, V L Area and V H The regions are located on different chains of a peptide that is an immunoglobulin fold protein.

[0292] In one technical solution, the first V L Area and the first V H region is oriented so that V L The three CDRs and V H The three CDRs in the region cooperate to selectively and / or specifically bind to an antigenic determinant on human CD38.

[0293] In another embodiment, the peptide comprises a second V L Same V L Area and the second and first V H Same V H Area, where the second V L Area and the second V H The regions are used together to selectively and / or specifically bind to antigenic determinants of human CD38.

[0294] The present invention provides a compound containing V as -003, -005 or -024. L V L CD38BP in the region.

[0295] In one embodiment, V of CD38BP L The region consists essentially of a sequence that has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% amino acid identity to the sequence set forth in SEQ ID No: 2, SEQ ID No: 12, or SEQ ID No: 22, respectively. In one embodiment, the CD38BP has at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% of the antigenic determinant binding properties of -003, -005, or -024, respectively.

[0296] The present invention provides a compound containing V as -003, -005 or -024. H V H CD38BP in the region.

[0297] In one embodiment, V of CD38BP HThe region consists essentially of a sequence that has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% amino acid identity to the sequence set forth in SEQ ID No: 7, SEQ ID No: 17, or SEQ ID No: 27, respectively. In one embodiment, the CD38BP has at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% of the antigenic determinant binding properties of -003, -005, or -024, respectively.

[0298] The present invention provides CD38BP comprising at least one CDR that is a functional variant of the CDR of -003 or -005 or -024.

[0299] In one embodiment, the CDRs of at least one peptide consist essentially of a sequence that is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% identical to the amino acid sequence of SEQ ID No: 3, SEQ ID No: 4, SEQ ID No: 5, SEQ ID No: 8, SEQ ID No: 9 or SEQ ID No: 10, or SEQ ID No: 13, SEQ ID No: 14, SEQ ID No: 15, SEQ ID No: 18, SEQ ID No: 19, or SEQ ID No: 20, or SEQ ID No: 23, SEQ ID No: 24, SEQ ID No: 25, SEQ ID No: 28, SEQ ID No: 29 or SEQ ID No: 30, respectively. In one embodiment, CD38BP has at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or at least about 95% of the antigenic determinant binding properties of -003 or -005 or -024, respectively.

[0300] In one embodiment, CD38BP has at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or at least about 95% of the affinity, potency or specificity of -003 or -005 or -024, respectively.

[0301] In one embodiment, CD38BP competes with -003 or -005 or -024 for binding to CD38.

[0302] In another embodiment, the competition is determined by ELISA as described in the Examples section. In another embodiment, the competition is determined by FACS as described in the Examples section.

[0303] In one technical solution, CD38BP specifically binds to a CD38 antigenic determinant, and its antigenic determinant also specifically binds to -003, -005, or -024.

[0304] In one embodiment, the affinity of CD38BP for binding to human CD38 is greater than that of -003, -005, or -024.

[0305] In one embodiment, CD38BP has substantially the same property of specifically binding to CD38 as -003, -005, or -024.

[0306] In one embodiment, the CD38BP is substantially free of other CD38 binding peptides.

[0307] In one embodiment, the CD38BP of the present invention is an antibody. In another embodiment, the CD38BP is a human antibody. In another embodiment, the CD38BP is a humanized antibody. In another embodiment, the CD38BP is a chimeric antibody.

[0308] In one embodiment, the antibody of the present invention is a monoclonal antibody.

[0309] In one embodiment, the antibody of the present invention is an IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM antibody. In another embodiment, the antibody is an IgG1 antibody. In another embodiment, the antibody is an IgG1κ antibody. In another embodiment, the antibody is an IgM antibody. In another embodiment, the antibody is an IgM1κ antibody.

[0310] In one embodiment, the antibody of the present invention is an antibody fragment or a single-chain antibody.

[0311] In one embodiment, CD38BP is glycosylated in eukaryotic cells.

[0312] In one embodiment, CD38BP further comprises a chelator linker for attaching a radioisotope.

[0313] In one embodiment, CD38BP is in substantially isolated form.

[0314] The present invention provides isolated nucleic acids encoding the CD38BP of the present invention.

[0315] The present invention provides an expression vector comprising a nucleic acid sequence encoding the CD38BP of the present invention.

[0316] In one technical solution, the expression vector contains V having sequence number SEQ ID No: 1. LNucleotide sequence, sequence number is SEQ ID No: 6 V H Nucleotide sequence, or V sequence number SEQ ID No: 1 L The nucleotide sequence and sequence number are SEQ ID No: 6 V H Nucleotide sequence.

[0317] In one technical solution, the expression vector contains V having a sequence number of SEQ ID No: 11. L Nucleotide sequence, sequence number is SEQ ID No: 16 V H Nucleotide sequence, or V sequence numbered SEQ ID No: 11 L The nucleotide sequence and sequence number are SEQ ID No: 16 V H Nucleotide sequence.

[0318] In one technical solution, the expression vector contains V having sequence number SEQ ID No: 21. L Nucleotide sequence, sequence number is SEQ ID No: 26 V H Nucleotide sequence, or V sequence numbered SEQ ID No: 21 L The nucleotide sequence and sequence number are SEQ ID No: 26 H Nucleotide sequence.

[0319] In another technical embodiment, the expression vector further contains a nucleotide sequence encoding the light chain, heavy chain, or light chain and heavy chain constant regions of a human antibody.

[0320] In another technical embodiment, the nucleotide sequence encoding the light chain, heavy chain, or light chain and heavy chain constant regions of a human antibody encodes an IgG1 antibody.

[0321] The present invention provides hybridomas capable of producing human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, wherein these sequences comprise the nucleotide sequence of the variable light chain region as set forth in SEQ ID No: 1, or conservative sequence variations thereof, and the nucleotide sequence of the variable heavy chain region as set forth in SEQ ID No: 6, or conservative sequence variations thereof. In one embodiment, the human light chain nucleic acid comprises the nucleotide sequence set forth in SEQ ID No: 1, and the human heavy chain nucleic acid comprises the nucleotide sequence set forth in SEQ ID No: 6.

[0322] The present invention provides a hybridoma capable of producing a human monoclonal anti-CD38 antibody having human light and heavy chain variable regions, comprising a human light chain variable amino acid sequence as set forth in SEQ ID No: 2 or a conservative sequence variation thereof, and a human heavy chain variable amino acid sequence as set forth in SEQ ID No: 7 or a conservative sequence variation thereof. In one embodiment, the human light chain variable region comprises the amino acid sequence as set forth in SEQ ID No: 2, and the human heavy chain variable region comprises the amino acid sequence as set forth in SEQ ID No: 7.

[0323] The present invention provides a transfectoma capable of producing a human monoclonal anti-CD38 antibody encoded by a nucleic acid encoding a human light chain variable region as in SEQ ID No: 1 or a conservative sequence variation thereof and a nucleic acid encoding a human heavy chain as in SEQ ID No: 6 or a conservative sequence variation thereof. In one embodiment, the human monoclonal anti-CD38 antibody is encoded by a nucleic acid encoding a human light chain variable region as in SEQ ID No: 1 and a nucleic acid encoding a human heavy chain as in SEQ ID No: 6.

[0324] The present invention provides a transfectoma capable of producing a human monoclonal anti-CD38 antibody having human light and heavy chain variable regions, wherein the variable regions comprise the amino acid sequence of the human light chain variable region as set forth in SEQ ID No: 2 or conservative sequence variations thereof, and the amino acid sequence of the human heavy chain variable region as set forth in SEQ ID No: 7 or conservative sequence variations thereof. In one embodiment, the human light chain comprises the amino acid sequence of the human light chain variable region as set forth in SEQ ID No: 2, and the human heavy chain comprises the amino acid sequence of the human heavy chain variable region as set forth in SEQ ID No: 7.

[0325] The present invention provides hybridomas capable of producing human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, wherein these sequences comprise the nucleotide sequence of the variable light chain region as set forth in SEQ ID No: 11, or conservative sequence variations thereof, and the nucleotide sequence of the variable heavy chain region as set forth in SEQ ID No: 16, or conservative sequence variations thereof. In one embodiment, the human light chain nucleic acid comprises the nucleotide sequence set forth in SEQ ID No: 11, and the human heavy chain nucleic acid comprises the nucleotide sequence set forth in SEQ ID No: 16.

[0326] The present invention provides hybridomas capable of producing human monoclonal anti-CD38 antibodies having human heavy and light chain variable regions, wherein these variable regions comprise the amino acid sequence of a human light chain variable region as set forth in SEQ ID No: 12, or conservative sequence variations thereof, and the amino acid sequence of a human heavy chain variable region as set forth in SEQ ID No: 17, or conservative sequence variations thereof. In one embodiment, the human light chain comprises the amino acid sequence of a human light chain variable region as set forth in SEQ ID No: 12, and the human heavy chain comprises the amino acid sequence of a human heavy chain variable region as set forth in SEQ ID No: 17.

[0327] The present invention provides a transfectoma capable of producing a human monoclonal anti-CD38 antibody encoded by a nucleic acid encoding a human light chain variable region as set forth in SEQ ID No: 11 or a conservative sequence variation thereof and a nucleic acid encoding a human heavy chain as set forth in SEQ ID No: 16 or a conservative sequence variation thereof. In one embodiment, the human monoclonal anti-CD38 antibody is encoded by a nucleic acid encoding a human light chain variable region as set forth in SEQ ID No: 11 and a nucleic acid encoding a human heavy chain as set forth in SEQ ID No: 16.

[0328] The present invention provides a transfectoma capable of producing a human monoclonal anti-CD38 antibody having human light and heavy chain variable regions, wherein the variable regions comprise the amino acid sequence of a human light chain variable region as set forth in SEQ ID No: 12 or conservative sequence variations thereof, and the amino acid sequence of a human heavy chain variable region as set forth in SEQ ID No: 17 or conservative sequence variations thereof. In one embodiment, the human light chain comprises the amino acid sequence of a human light chain variable region as set forth in SEQ ID No: 12, and the human heavy chain comprises the amino acid sequence of a human heavy chain variable region as set forth in SEQ ID No: 17.

[0329] The present invention provides hybridomas capable of producing human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, wherein these sequences comprise the nucleotide sequence of the variable light chain region as set forth in SEQ ID No: 21, or conservative sequence variations thereof, and the nucleotide sequence of the variable heavy chain region as set forth in SEQ ID No: 26, or conservative sequence variations thereof. In one embodiment, the human light chain nucleic acid comprises the nucleotide sequence set forth in SEQ ID No: 21, and the human heavy chain nucleic acid comprises the nucleotide sequence set forth in SEQ ID No: 26.

[0330] The present invention provides a transfectoma capable of producing a human monoclonal anti-CD38 antibody having human heavy and light chain variable regions, wherein the variable regions comprise the amino acid sequence of a human light chain variable region as set forth in SEQ ID No: 22, or conservative sequence variations thereof, and the amino acid sequence of a human heavy chain variable region as set forth in SEQ ID No: 27, or conservative sequence variations thereof. In one embodiment, the human light chain variable region comprises the amino acid sequence of a human light chain variable region as set forth in SEQ ID No: 22, and the human heavy chain variable region comprises the amino acid sequence of a human heavy chain variable region as set forth in SEQ ID No: 27.

[0331] The present invention provides a transfectoma capable of producing a human monoclonal anti-CD38 antibody encoded by a nucleic acid encoding a human light chain variable region as in SEQ ID No: 21 or a conservative sequence variation thereof and a nucleic acid encoding a human heavy chain as in SEQ ID No: 26 or a conservative sequence variation thereof. In one embodiment, the human monoclonal anti-CD38 antibody is encoded by a nucleic acid encoding a human light chain variable region as in SEQ ID No: 21 and a nucleic acid encoding a human heavy chain as in SEQ ID No: 26.

[0332] The present invention provides a transfectoma capable of producing a human monoclonal anti-CD38 antibody having human light and heavy chain variable regions, wherein the variable regions comprise the amino acid sequence of a human light chain variable region as set forth in SEQ ID No: 22 or conservative sequence variations thereof, and the amino acid sequence of a human heavy chain variable region as set forth in SEQ ID No: 27 or conservative sequence variations thereof. In one embodiment, the human light chain comprises the amino acid sequence of a human light chain variable region as set forth in SEQ ID No: 22, and the human heavy chain comprises the amino acid sequence of a human heavy chain variable region as set forth in SEQ ID No: 27.

[0333] The present invention provides eukaryotic or prokaryotic host cells that can produce the CD38BP of the present invention.

[0334] The present invention provides a eukaryotic or prokaryotic host cell containing the expression vector of the present invention.

[0335] The present invention provides transgenic animals or plants containing nucleic acids encoding human heavy chains and human light chains, wherein the animals or plants produce detectable amounts of the CD38BP of the present invention.

[0336] The present invention provides immunoconjugates of the CD38BP of the present invention linked to an adenoviral agent, a radioisotope, or a drug. In one embodiment, the peptide is a monovalent IgM antibody linked to an adenoviral agent, a radioisotope, or a drug.

[0337] The present invention provides bispecific or multispecific molecules comprising the CD38BP of the present invention and having binding specificity for human effector cells. In one embodiment, the binding specificity for human effector cells refers to binding specificity for CD3, CD4, CD138, IL-15R, membrane-bound or receptor-bound TNF-α, human Fc receptor, or membrane-bound or receptor-bound IL-15.

[0338] The present invention provides an anti-idiotypic antibody that binds to the CD38BP of the present invention. The present invention provides use of the anti-idiotypic antibody of the present invention in detecting the level of human monoclonal antibodies against CD38 in a sample.

[0339] The following is a list of technical solutions selected by the present invention.

[0340] Technical Solution 1: An antibody that binds to human CD38 is encoded by a human light chain and a human heavy chain nucleic acid whose variable regions contain the nucleotide sequences shown in SEQ ID No: 1 and SEQ ID No: 6, respectively, or conservative sequence changes thereof.

[0341] Technical Solution 2: An antibody that binds to human CD38, encoded by human light chain and human heavy chain nucleic acids whose variable regions contain the nucleotide sequences shown in SEQ ID No: 1 and SEQ ID No: 6, respectively.

[0342] Technical Solution 3: An antibody that binds to human CD38, encoded by a human light chain and a human heavy chain nucleic acid whose variable regions contain the nucleotide sequences shown in SEQ ID No: 11 and SEQ ID No: 16, respectively, or conservative sequence changes thereof.

[0343] Technical Solution 4: An antibody that binds to human CD38, encoded by human light chain and human heavy chain nucleic acids whose variable regions contain the nucleotide sequences shown in SEQ ID No: 11 and SEQ ID No: 26, respectively.

[0344] Technical Solution 5: An antibody that binds to human CD38, encoded by a human light chain and a human heavy chain nucleic acid whose variable regions contain the nucleotide sequences shown in SEQ ID No: 21 and SEQ ID No: 26, respectively, or conservative sequence changes thereof.

[0345] Technical Solution 6: An antibody that binds to human CD38, encoded by human light chain and human heavy chain nucleic acids whose variable regions contain the nucleotide sequences shown in SEQ ID No: 21 and SEQ ID No: 26, respectively.

[0346] Technical Solution 7: A peptide that competes with the antibody described in Technical Solution 2 for binding to CD38.

[0347] Technical Solution 8: The peptide as described in Technical Solution 7, wherein the competition is determined by ELISA described in Example 8 or 9 of the specification.

[0348] Technical Solution 9: The peptide as described in Technical Solution 7, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

[0349] Technical Solution 10: A peptide that specifically binds to a CD38 antigenic determinant, wherein the antigenic determinant can also be specifically bound by the antibody described in Technical Solution 2.

[0350] Technical Solution 11: A peptide having substantially the same binding properties for specifically binding to human CD38 as the antibody described in Technical Solution 2.

[0351] Technical solution 12: containing V mainly composed of SEQ ID No: 3 L CDR1 peptide.

[0352] Technical solution 13: containing V mainly composed of SEQ ID No: 4 L CDR2 peptide.

[0353] Technical solution 14: containing V mainly composed of SEQ ID No: 5 L CDR3 peptides.

[0354] Technical Solution 15: The peptide as described in Technical Solution 14, which also contains V mainly composed of SEQ ID No: 3 L CDR1.

[0355] Technical Solution 16: The peptide as described in Technical Solution 14, which also contains V mainly composed of SEQ ID No: 4 L CDR2.

[0356] Technical Solution 17: The peptide as described in Technical Solution 16, which also contains V mainly composed of SEQ ID No: 3 L CDR1.

[0357] Technical solution 18: containing V mainly composed of SEQ ID No: 8 H CDR1 peptide.

[0358] Technical solution 19: containing V mainly composed of SEQ ID No: 9 H CDR2 peptide.

[0359] Technical solution 20: containing V mainly composed of SEQ ID No: 10 H CDR3 peptides.

[0360] Technical Solution 21: The peptide according to Technical Solution 20, which also contains V mainly composed of SEQ ID No: 8 H CDR1.

[0361] Technical Solution 22: The peptide as described in Technical Solution 20, which also contains V mainly composed of SEQ ID No: 9 H CDR2.

[0362] Technical Solution 23: The peptide as described in Technical Solution 22, which also contains V mainly composed of SEQ ID No: 8 H CDR1.

[0363] Technical Solution 24: Peptide containing

[0364] (a) Contains 3 V L The first V of CDRs L regions, each of which consists essentially of SEQ ID No: 3, SEQ ID No: 4, and SEQ ID No: 5; and

[0365] (b) Contains 3 V H The first V of CDRsH regions, each of which consists essentially of SEQ ID No: 8, SEQ ID No: 9, and SEQ ID No: 10.

[0366] Technical Solution 25: A peptide as described in Technical Solution 24, wherein V L Area and V H The regions are located on the same chain of the peptide.

[0367] Technical Solution 26: The peptide as described in Technical Solution 25, wherein V L Area and V H The zones are separated by flexible connectors.

[0368] Technical Solution 27: A peptide as described in Technical Solution 24, wherein V L Area and V H The regions are located on different chains of the peptide.

[0369] Technical Solution 28: The peptide as described in Technical Solution 27, wherein V L Area and V H The regions are located on different chains of a peptide that is an immunoglobulin fold protein.

[0370] Technical Solution 29: A peptide as described in any one of Technical Solutions 24 to 28, wherein the first V L Area and the first V H region is oriented so that V L The three CDRs and V H The three CDRs in the region cooperate to selectively and / or specifically bind to an antigenic determinant on human CD38.

[0371] Technical Solution 30: A peptide as described in Technical Solution 29, wherein the peptide contains a second V L Same V L Area and the second and first V H Same V H Area, where the second V L Area and the second V H The two regions act synergistically to selectively and / or specifically bind to antigenic determinants on human CD38.

[0372] Technical solution 31: containing the antibody V as technical solution 2 L V-domain functional variants L region of peptides.

[0373] Technical Solution 32: A peptide as described in Technical Solution 31, wherein V LThe region consists essentially of a sequence that is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% identical to the amino acid sequence set forth in SEQ ID No: 2.

[0374] Technical solution 33: containing the antibody V as technical solution 2 H V-domain functional variants H region of peptides.

[0375] Technical Solution 34: A peptide as described in Technical Solution 33, wherein V of the peptide H The region consists essentially of a sequence that is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% identical to the amino acid sequence set forth in SEQ ID No:7.

[0376] Technical Solution 35: A peptide containing at least one CDR that is a functional variant of the antibody CDR in Technical Solution 2.

[0377] Technical Solution 36: A peptide as described in Technical Solution 35, wherein the CDRs of at least one peptide are mainly composed of a sequence that has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% amino acid identity with the sequence described in SEQ ID No: 3, SEQ ID No: 4, SEQ ID No: 5, SEQ ID No: 8, SEQ ID No: 9 or SEQ ID No: 10.

[0378] Technical Solution 37: A peptide as described in any one of Technical Solutions 31 to 36, wherein the peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% of the antigen determinant binding properties of the antibody in Technical Solution 2.

[0379] Technical Solution 38: A peptide as described in any one of Technical Solutions 31 to 36, wherein the peptide has an affinity, strength or specificity of at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% of the antibody in Technical Solution 2.

[0380] Technical Solution 39: The peptide as described in any one of Technical Solutions 12 to 38, which specifically binds to human CD38.

[0381] Technical Solution 40: The peptide according to any one of Technical Solutions 12 to 39, which competes with the antibody according to Technical Solution 2 for binding to CD38.

[0382] Technical Solution 41: A peptide as described in Technical Solution 40, wherein the competition is determined by ELISA described in Example 8 or 9 of the specification.

[0383] Technical Solution 42: The peptide as described in Technical Solution 7, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

[0384] Technical Solution 43: The peptide as described in Technical Solution 39, which specifically binds to the CD38 antigenic determinant, and the antigenic determinant also specifically binds to the antibody as described in Technical Solution 2.

[0385] Technical Solution 44: The peptide as described in any one of Technical Solutions 39 to 43, wherein the binding affinity of the peptide to human CD38 is greater than that of the antibody as described in Technical Solution 2.

[0386] Technical Solution 45: The peptide as described in any one of Technical Solutions 39 to 43, wherein the peptide has substantially the same specific CD38 binding properties as the antibody as described in Technical Solution 2.

[0387] Technical Solution 46: The peptide according to any one of Technical Solutions 39 to 45, wherein the CD38-binding peptide does not substantially contain other CD38-binding peptides.

[0388] Technical Solution 47: A peptide that binds to human CD38 (SEQ ID No: 31), the degree of binding of which to a mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue is different from the degree of binding of the peptide to human CD38 (SEQ ID No: 31).

[0389] Technical solution 48: The peptide as described in technical solution 47, wherein the peptide binds to an EC in which the mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 50% of.

[0390] Technical solution 49: The peptide as described in technical solution 48, wherein the peptide binds to an EC in which the mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 10% of.

[0391] Technical solution 50: The peptide as described in technical solution 49, wherein the peptide binds to an EC in which the mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 5% of.

[0392] Technical solution 51: The peptide as described in technical solution 50, wherein the peptide binds to an EC in which the mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 1%.

[0393] Technical Solution 52: A peptide that binds to human CD38 (SEQ ID No: 31), the degree of binding of which to a mutated human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue is different from the degree of binding of the peptide to human CD38 (SEQ ID No: 31).

[0394] Technical solution 53: The peptide as described in technical solution 52, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 50% of.

[0395] Technical solution 54: The peptide as described in technical solution 53, wherein the peptide binds to an EC in which the mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 10% of.

[0396] Technical Solution 55: The degree of binding of the peptide in any one of Technical Solutions 47 to 51 to a mutated human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue is different from the degree of binding to human CD38 (SEQ ID No: 31).

[0397] Technical solution 56: The peptide as described in technical solution 55, wherein the peptide binds to an EC in which the mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50The peptide binds to human CD38 (SEQ ID No: 31) 50 50% of.

[0398] Technical solution 57: The peptide as described in technical solution 56, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) is substituted. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 10% of.

[0399] Technical Solution 58: A peptide as described in any one of Technical Solutions 47 to 57, wherein the peptide binds to a mutated human CD38 (SEQ ID No: 32) in which the threonine residue at position 237 is replaced by an alanine residue, and the degree of binding is the same as its binding to human CD38 (SEQ ID No: 31).

[0400] Technical solution 59: The peptide as described in technical solution 58, wherein the peptide binds to the EC of human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 75% of.

[0401] Technical solution 60: The peptide as described in technical solution 59, wherein the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 85% of.

[0402] Technical solution 61: A peptide as described in technical solution 60, wherein the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 90% of.

[0403] Technical solution 62: The peptide as described in technical solution 61, wherein the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 95% of.

[0404] Technical Solution 63: A peptide that competes with the antibody as described in Technical Solution 4 for binding to CD38.

[0405] Technical Solution 64: A peptide as described in Technical Solution 63, wherein the competition is determined by ELISA described in Example 8 or 9 of the specification.

[0406] Technical Solution 65: The peptide as described in Technical Solution 63, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

[0407] Technical Solution 66: A peptide that specifically binds to a CD38 antigenic determinant, which also specifically binds to the antibody described in Technical Solution 4.

[0408] Technical Solution 67: A peptide having substantially the same specific binding properties for binding to human CD38 as the antibody described in Technical Solution 4.

[0409] Technical solution 68: containing V mainly composed of SEQ ID No: 13 L CDR1 peptide.

[0410] Technical solution 69: containing V mainly composed of SEQ ID No: 14 L CDR2 peptide.

[0411] Technical solution 70: containing V mainly composed of SEQ ID No: 15 L CDR3 peptides.

[0412] Technical solution 71: The peptide as described in technical solution 70, which also contains V mainly composed of SEQ ID No: 13 L CDR1.

[0413] Technical solution 72: The peptide as described in technical solution 70, which also contains V mainly composed of SEQ ID No: 14 L CDR2.

[0414] Technical solution 73: The peptide as described in technical solution 72, which also contains V mainly composed of SEQ ID No: 13 L CDR1.

[0415] Technical solution 74: containing V mainly composed of SEQ ID No: 18 H CDR1 peptide.

[0416] Technical solution 75: containing V mainly composed of SEQ ID No: 19 H CDR2 peptide.

[0417] Technical solution 76: containing V mainly composed of SEQ ID No: 20 H CDR3 peptides.

[0418] Technical solution 77: A peptide as described in technical solution 76, which also contains V mainly composed of SEQ ID No: 18 H CDR1.

[0419] Technical solution 78: The peptide as described in technical solution 76, which also contains V mainly composed of SEQ ID No: 19 H CDR2.

[0420] Technical solution 79: The peptide as described in technical solution 78, which also contains V mainly composed of SEQ ID No: 18 H CDR1.

[0421] Technical Solution 80: Peptide, containing

[0422] (a) Contains 3 V L The first V of CDRs L regions, each of which consists essentially of SEQ ID No: 13, SEQ ID No: 14, and SEQ ID No: 15; and

[0423] (b) Contains 3 V H The first V of CDRs H regions, each of which consists essentially of SEQ ID No: 18, SEQ ID No: 19, and SEQ ID No: 20.

[0424] Technical Solution 81: A peptide as described in Technical Solution 80, wherein V L Area and V H The regions are located on the same chain of the peptide.

[0425] Technical Solution 82: A peptide as described in Technical Solution 81, wherein V L Area and V H The zones are separated by flexible connectors.

[0426] Technical solution 83: A peptide as described in technical solution 80, wherein V L Area and V H The regions are located on different chains of the peptide.

[0427] Technical solution 84: A peptide as described in technical solution 83, wherein V L Area and V H The regions are located on different chains of a peptide that is an immunoglobulin fold protein.

[0428] Technical solution 85: A peptide as described in any one of technical solutions 80 to 84, wherein the first V L Area and the first V H region is oriented so that V L The three CDRs and V H The three CDRs in the region cooperate to selectively and / or specifically bind to an antigenic determinant on human CD38.

[0429] Technical Solution 86: A peptide as described in Technical Solution 85, wherein the peptide contains a second V L Same V L Area and the second and first V H Same V H Area, where the second V L Area and the second V H The two regions act synergistically to selectively and / or specifically bind to antigenic determinants on human CD38.

[0430] Technical solution 87: Containing the antibody V as technical solution 4 L V-domain functional variants L region of peptides.

[0431] Technical solution 88: A peptide as described in technical solution 87, wherein V of the peptide L The region consists essentially of a sequence that is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% identical to the amino acid sequence set forth in SEQ ID No: 12.

[0432] Technical solution 89: containing the antibody V as technical solution 4 H V-domain functional variants H region of peptides.

[0433] Technical Solution 90: A peptide as described in Technical Solution 89, wherein V H The region consists essentially of a sequence that is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% identical to the amino acid sequence set forth in SEQ ID No: 17.

[0434] Technical Solution 91: A peptide comprising at least one CDR that is a functional variant of the antibody CDR of Technical Solution 4.

[0435] Technical Solution 92: A peptide as described in Technical Solution 91, wherein the CDRs of at least one peptide are mainly composed of a sequence that has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% amino acid identity with the sequence described in SEQ ID No: 13, SEQ ID No: 14, SEQ ID No: 15, SEQ ID No: 18, SEQ ID No: 19 or SEQ ID No: 20.

[0436] Technical Solution 93: A peptide as described in any one of Technical Solutions 87 to 92, wherein the peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% of the antigen determinant binding properties of the antibody in Technical Solution 4.

[0437] Technical Solution 94: A peptide as described in any one of Technical Solutions 87 to 92, wherein the peptide has an affinity, strength or specificity of at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% of the antibody in Technical Solution 4.

[0438] Technical Solution 95: The peptide as described in any one of Technical Solutions 68 to 94, which specifically binds to human CD38.

[0439] Technical Solution 96: A peptide as described in any one of Technical Solutions 68 to 95, which competes with the antibody as described in Technical Solution 4 for binding to CD38.

[0440] Technical Solution 97: A peptide as described in Technical Solution 96, wherein the competition is determined by the ELISA described in Example 8 or 9 of the specification.

[0441] Technical Solution 98: A peptide as described in Technical Solution 96, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

[0442] Technical Solution 99: The peptide as described in Technical Solution 95, which specifically binds to the CD38 antigenic determinant, and the antigenic determinant also specifically binds to the antibody as described in Technical Solution 4.

[0443] Technical Solution 100: The peptide as described in any one of Technical Solutions 95 to 99, wherein the binding affinity of the peptide to human CD38 is greater than that of the antibody as described in Technical Solution 4.

[0444] Technical Solution 101: The peptide as described in any one of Technical Solutions 95 to 99, wherein the peptide has substantially the same specific CD38 binding properties as the antibody as described in Technical Solution 4.

[0445] Technical Solution 102: The peptide as described in any one of Technical Solutions 95 to 101, wherein the CD38-binding peptide does not substantially contain other CD38-binding peptides.

[0446] Technical Solution 103: A peptide as described in any one of Technical Solutions 63 to 102, which binds to human CD38 (SEQ ID No: 31), but does not bind to a mutated human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue to the same extent as it binds to human CD38 (SEQ ID No: 31).

[0447] Technical solution 104: The peptide as described in technical solution 103, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 50% of.

[0448] Technical solution 105: The peptide according to technical solution 104, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) is substituted. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 10% of.

[0449] Technical solution 106: The peptide according to technical solution 105, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) is substituted. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 5% of.

[0450] Technical solution 107: The peptide according to technical solution 106, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) is substituted. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 1%.

[0451] Technical Solution 108: A peptide that binds to human CD38 (SEQ ID No: 31), the degree of binding of which to a mutated human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue is different from the degree of binding of the peptide to human CD38 (SEQ ID No: 31).

[0452] Technical solution 109: The peptide according to technical solution 108, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) is substituted. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 50% of.

[0453] Technical solution 110: The peptide according to technical solution 109, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) is substituted. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 10% of.

[0454] Technical Solution 111: The degree of binding of the peptide in any one of Technical Solutions 103 to 107 to a mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue is different from the degree of binding to human CD38 (SEQ ID No: 31).

[0455] Technical solution 112: The peptide as described in technical solution 111, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) is substituted. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 50% of.

[0456] Technical solution 113: The peptide according to technical solution 112, wherein the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and the mutant human CD38 (SEQ ID No: 34) is substituted. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 10% of.

[0457] Technical Solution 114: A peptide as described in any one of Technical Solutions 103 to 113, wherein the peptide binds to a mutated human CD38 (SEQ ID No: 32) in which the threonine residue at position 237 is replaced by an alanine residue, and the degree of binding is the same as its binding to human CD38 (SEQ ID No: 31).

[0458] Technical solution 115: A peptide as described in technical solution 114, wherein the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 75% of.

[0459] Technical solution 116: The peptide according to technical solution 115, wherein the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 85% of.

[0460] Technical solution 117: The peptide according to technical solution 116, wherein the peptide binds to an EC of a mutated human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 90% of.

[0461] Technical solution 118: The peptide as described in technical solution 117, wherein the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 95% of.

[0462] Technical Solution 119: A peptide that competes with the antibody as described in Technical Solution 6 for binding to CD38.

[0463] Technical solution 120: A peptide as described in technical solution 119, wherein the competition is determined by ELISA described in Example 8 or 9 of the specification.

[0464] Technical Solution 121: A peptide as described in Technical Solution 119, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

[0465] Technical Solution 122: A peptide that specifically binds to a CD38 antigenic determinant, which also specifically binds to the antibody described in Technical Solution 6.

[0466] Technical Solution 123: A peptide having substantially the same specific binding properties for binding to human CD38 as the antibody according to Technical Solution 6.

[0467] Technical solution 124: containing V mainly composed of SEQ ID No: 23 L CDR1 peptide.

[0468] Technical solution 125: containing V mainly composed of SEQ ID No: 24 L CDR2 peptide.

[0469] Technical solution 126: containing V mainly composed of SEQ ID No: 25 L CDR3 peptides.

[0470] Technical solution 127: The peptide according to technical solution 126, which also contains V mainly composed of SEQ ID No: 23 L CDR1.

[0471] Technical solution 128: The peptide according to technical solution 126, which also contains V mainly composed of SEQ ID No: 24 L CDR2.

[0472] Technical solution 129: The peptide according to technical solution 128, which also contains V mainly composed of SEQ ID No: 23 L CDR1.

[0473] Technical solution 130: containing V mainly composed of SEQ ID No: 28 H CDR1 peptide.

[0474] Technical solution 131: containing V mainly composed of SEQ ID No: 29 H CDR2 peptide.

[0475] Technical solution 132: containing V mainly composed of SEQ ID No: 30 H CDR3 peptides.

[0476] Technical Solution 133: The peptide according to Technical Solution 132, which also contains V mainly composed of SEQ ID No: 28 H CDR1.

[0477] Technical solution 134: The peptide according to technical solution 132, which also contains V mainly composed of SEQ ID No: 29 H CDR2.

[0478] Technical solution 135: The peptide according to technical solution 134, which also contains V mainly composed of SEQ ID No: 28 H CDR1.

[0479] Technical Solution 136: Peptide, containing

[0480] (a) Contains 3 V L The first V of CDRs L regions, each of which consists essentially of SEQ ID No: 23, SEQ ID No: 24, and SEQ ID No: 25; and

[0481] (b) Contains 3 V H The first V of CDRs H regions, each of which consists essentially of SEQ ID No: 28, SEQ ID No: 29, and SEQ ID No: 30.

[0482] Technical Solution 137: A peptide as described in Technical Solution 136, wherein V L Area and V H The regions are located on the same chain of the peptide.

[0483] Technical Solution 138: A peptide as described in Technical Solution 137, wherein V L Area and V H The zones are separated by flexible connectors.

[0484] Technical Solution 139: A peptide as described in Technical Solution 136, wherein V L Area and V H The regions are located on different chains of the peptide.

[0485] Technical Solution 140: A peptide as described in Technical Solution 139, wherein V L Area and V H The regions are located on different chains of a peptide that is an immunoglobulin fold protein.

[0486] Technical Solution 141: A peptide as described in any one of Technical Solutions 136 to 140, wherein the first V L Area and the first V H region is oriented so that V L The three CDRs and V H The three CDRs in the region cooperate to selectively and / or specifically bind to an antigenic determinant on human CD38.

[0487] Technical Solution 142: A peptide as described in Technical Solution 141, wherein the peptide contains a second V L Same V L Area and the second and first V H Same V H Area, where the second V L Area and the second V H The two regions act synergistically to selectively and / or specifically bind to antigenic determinants on human CD38.

[0488] Technical solution 143: containing the antibody V as technical solution 6 L V-domain functional variants L region of peptides.

[0489] Technical Solution 144: A peptide as described in Technical Solution 143, wherein V of the peptide L The region consists essentially of a sequence that is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% identical to the amino acid sequence set forth in SEQ ID No: 22.

[0490] Technical solution 145: containing the antibody V as technical solution 6 H V-domain functional variants H region of peptides.

[0491] Technical Solution 146: A peptide as described in Technical Solution 145, wherein V of the peptide H The region consists essentially of a sequence that is at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% identical to the amino acid sequence set forth in SEQ ID No: 27.

[0492] Technical Solution 147: A peptide comprising at least one CDR that is a functional variant of the antibody CDR of Technical Solution 6.

[0493] Technical Solution 148: A peptide as described in Technical Solution 147, wherein the CDRs of at least one peptide are mainly composed of a sequence that has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% amino acid identity with the sequence described in SEQID No: 23, SEQ ID No: 24, SEQ ID No: 25, SEQ ID No: 28, SEQ ID No: 29 or SEQ ID No: 30.

[0494] Technical Solution 149: A peptide as described in any one of Technical Solutions 143 to 148, wherein the peptide has at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% of the antigen determinant binding properties of the antibody in Technical Solution 6.

[0495] Technical solution 150: A peptide as described in any one of technical solutions 143 to 148, wherein the peptide has an affinity, strength or specificity of at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or at least about 95% of the antibody in technical solution 6.

[0496] Technical Solution 151: The peptide as described in any one of Technical Solutions 124 to 150, which specifically binds to human CD38.

[0497] Technical Solution 152: A peptide as described in any one of Technical Solutions 124 to 151, which competes with the antibody as described in Technical Solution 6 for binding to CD38.

[0498] Technical Solution 153: A peptide as described in Technical Solution 152, wherein the competition is determined by ELISA described in Example 8 or 9 of the specification.

[0499] Technical solution 154: The peptide as described in technical solution 152, wherein the competition is determined by the cross-inhibition method described in Example 7 of the specification.

[0500] Technical Solution 155: The peptide as described in Technical Solution 151, which specifically binds to the CD38 antigenic determinant, and the antigenic determinant also specifically binds to the antibody as described in Technical Solution 6.

[0501] Technical Solution 156: The peptide according to any one of Technical Solutions 151 to 155, wherein the binding affinity of the peptide to human CD38 is greater than that of the antibody according to Technical Solution 6.

[0502] Technical Solution 157: The peptide as described in any one of Technical Solutions 151 to 155, wherein the peptide has substantially the same specific CD38 binding properties as the antibody as described in Technical Solution 6.

[0503] Technical solution 158: The peptide according to any one of technical solutions 151 to 157, wherein the CD38-binding peptide does not substantially contain other CD38-binding peptides.

[0504] Technical Solution 159: The peptide as described in any one of Technical Solutions 1 to 158, wherein the peptide is not an antagonist of CD38.

[0505] Technical Solution 160: The peptide as described in any one of Technical Solutions 1 to 159, wherein the peptide does not induce significant proliferation of peripheral blood mononuclear cells.

[0506] Technical Solution 161: The peptide as described in any one of Technical Solutions 1 to 160, wherein the peptide does not induce significant release of IL-6 from human monocytes or peripheral blood mononuclear cells.

[0507] Technical Solution 162: The peptide as described in any one of Technical Solutions 1 to 161, wherein the peptide does not induce human T cells or peripheral blood mononuclear cells to release detectable IFN-γ.

[0508] Technical Solution 163: The peptide as described in any one of Technical Solutions 1 to 162, wherein the peptide is an antibody.

[0509] Technical solution 164: The antibody as described in any one of technical solutions 1 to 6 or 163, wherein the antibody is a human antibody.

[0510] Technical Solution 165: The antibody as described in any one of Technical Solutions 1 to 6 or 163, wherein the antibody is a humanized antibody.

[0511] Technical Solution 166: The antibody as described in any one of Technical Solutions 1 to 6 or 163, wherein the antibody is a chimeric antibody.

[0512] Technical Solution 167: The antibody as described in any one of Technical Solutions 1 to 6 or 163, wherein the antibody is a monoclonal antibody.

[0513] Technical Solution 168: The antibody as described in any one of Technical Solutions 1 to 6 or 163, characterized in that it is an IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE or IgM antibody.

[0514] Technical Solution 169: The antibody as described in Technical Solution 168 is characterized in that it is an IgG1 antibody.

[0515] Technical Solution 170: The antibody as described in Technical Solution 169 is characterized in that it is an IgG1,κ antibody.

[0516] Technical Solution 171: The antibody as described in Technical Solution 168 is characterized in that it is an IgM antibody.

[0517] Technical solution 172: The antibody as described in technical solution 171, wherein the antibody is an IgM1κ antibody.

[0518] Technical Solution 173: The peptide as described in any one of Technical Solutions 2 to 172, wherein the peptide is glycosylated in eukaryotic cells.

[0519] Technical solution 174: The antibody as described in any one of technical solutions 1 to 6 or 163, which is an antibody fragment or a single-chain antibody.

[0520] Technical Solution 175: The peptide or antibody as described in any one of Technical Solutions 1 to 174, further comprising a chelate linker for linking a radioactive isotope.

[0521] Technical Solution 176: The peptide as described in any one of Technical Solutions 1 to 175, which is in substantially isolated form.

[0522] Technical solution 177: An isolated nucleic acid encoding the peptide as described in any one of Technical Solutions 1 to 175.

[0523] Technical solution 178: An expression vector containing a nucleic acid encoding the peptide as described in any one of Technical Solutions 1 to 175.

[0524] Technical solution 179: containing V in SEQ ID No: 1 L Nucleotide sequence, V in SEQ ID No: 6 H Nucleotide sequence or V in SEQ ID No: 1 L Nucleotide sequence and V in SEQ ID No: 6 H Nucleotide sequence expression vector.

[0525] Technical solution 180: containing V in SEQ ID No: 11 L Nucleotide sequence, V in SEQ ID No: 16 H Nucleotide sequence or V in SEQ ID No: 11 L Nucleotide sequence and V in SEQ ID No: 16 H Nucleotide sequence expression vector.

[0526] Technical Solution 181: The expression vector as described in Technical Solution 179 or Technical Solution 180 further includes a nucleotide sequence encoding the constant region of the light chain, heavy chain, or light chain and heavy chain of a human antibody.

[0527] Technical Solution 182: An expression vector as described in Technical Solution 181, wherein the nucleotide sequence encoding the light chain, heavy chain, or the constant region of the light chain and heavy chain of a human antibody encodes an IgG1 antibody.

[0528] Technical Solution 183: A hybridoma that can produce human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, wherein these sequences include the nucleotide sequence of the variable light chain region as in SEQ ID No: 1 or its conservative sequence changes, and the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 6 or its conservative sequence changes.

[0529] Technical Solution 184: The hybridoma as described in Technical Solution 183, wherein the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 1, and the human heavy chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 6.

[0530] Technical Solution 185: A hybridoma that can produce human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, these sequences include the nucleotide sequence of the variable light chain region as in SEQ ID No: 2 or its conservative sequence changes, and the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 7 or its conservative sequence changes.

[0531] Technical Solution 186: The hybridoma as described in Technical Solution 185, wherein the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 2, and the human heavy chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 7.

[0532] Technical Solution 187: A transfectoma that can produce a human monoclonal anti-CD38 antibody encoded by a nucleic acid of a human light chain variable region as in SEQ ID No: 1 or a conservative sequence change thereof and a nucleic acid of a human heavy chain as in SEQ ID No: 6 or a conservative sequence change thereof.

[0533] Technical Solution 188: The transfectoma as described in Technical Solution 187, wherein the human monoclonal anti-CD38 antibody is encoded by the human light chain variable region nucleic acid as in SEQ ID No: 1 and the human heavy chain nucleic acid as in SEQ ID No: 6.

[0534] Technical Solution 189: A transfectoma capable of producing a human monoclonal anti-CD38 antibody having human light chain and heavy chain variable regions, wherein these variable regions include the amino acid sequence of the human light chain variable region as in SEQ ID No: 2 or a conservative sequence change thereof, and the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 7 or a conservative sequence change thereof.

[0535] Technical Solution 190: In the transfectoma as described in Technical Solution 189, the human light chain contains the human light chain variable region amino acid sequence as in SEQ ID No: 2, and the human heavy chain contains the human heavy chain variable region amino acid sequence as in SEQ ID No: 7.

[0536] Technical Solution 191: A hybridoma that can produce human monoclonal anti-CD38 antibodies encoded by human light chain and human heavy chain nucleic acids, wherein these sequences include the nucleotide sequence of the variable light chain region as in SEQ ID No: 11 or its conservative sequence changes, and the nucleotide sequence of the variable heavy chain region as in SEQ ID No: 16 or its conservative sequence changes.

[0537] Technical Solution 192: The hybridoma as described in Technical Solution 191, wherein the human light chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 11, and the human heavy chain nucleic acid contains the nucleotide sequence shown in SEQ ID No: 16.

[0538] Technical Solution 193: A hybridoma capable of producing human monoclonal anti-CD38 antibodies having human heavy and light chain variable regions, wherein these variable regions include the amino acid sequence of the human light chain variable region as in SEQ ID No: 12 or a conservative sequence change thereof, and the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 17 or a conservative sequence change thereof.

[0539] Technical Solution 194: The hybridoma as described in Technical Solution 193, wherein the human light chain variable region contains the amino acid sequence as in SEQ ID No: 12, and the human heavy chain variable region contains the amino acid sequence as in SEQ ID No: 17.

[0540] Technical Solution 195: A transfectoma that can produce a human monoclonal anti-CD38 antibody encoded by a nucleic acid of a human light chain variable region as in SEQ ID No: 11 or a conservative sequence change thereof and a nucleic acid of a human heavy chain as in SEQ ID No: 16 or a conservative sequence change thereof.

[0541] Technical Solution 196: The hybridoma as described in Technical Solution 195, wherein the human monoclonal anti-CD38 antibody is encoded by the human light chain variable region nucleic acid as in SEQ ID No: 11 and the human heavy chain nucleic acid as in SEQ ID No: 16.

[0542] Technical Solution 197: A transfectoma that can produce a human monoclonal anti-CD38 antibody having human light chain and heavy chain variable regions, wherein these variable regions include the amino acid sequence of the human light chain variable region as in SEQ ID No: 12 or a conservative sequence change thereof, and the amino acid sequence of the human heavy chain variable region as in SEQ ID No: 17 or a conservative sequence change thereof.

[0543] Technical Solution 198: The hybridoma as described in Technical Solution 197, wherein the human light chain contains the human light chain variable region amino acid sequence as in SEQ ID No: 12, and the human heavy chain contains the human heavy chain variable region amino acid sequence as in SEQ ID No: 17.

[0544] Technical Solution 199: A eukaryotic or prokaryotic host cell that can produce the peptide as described in any one of Technical Solutions 1 to 175.

[0545] Technical Solution 200: A eukaryotic or prokaryotic host cell containing the expression vector as described in Technical Solution 178.

[0546] Technical Solution 201: A transgenic non-human animal or plant containing nucleic acids encoding human heavy chains and human light chains, wherein the animal or plant produces a detectable amount of a peptide as described in any one of Technical Solutions 1 to 175.

[0547] Technical solution 202: An immunoconjugate comprising a peptide as described in any one of technical solutions 1 to 174 linked to a cytotoxic agent, a radioisotope or a drug.

[0548] Technical Solution 203: An immunoconjugate comprising a peptide as described in any one of Technical Solutions 1 to 55 or Technical Solutions 171 to 174 linked to a cytotoxic agent, radioisotope or drug, wherein the peptide is a monovalent IgM antibody linked to a cytotoxic agent, radioisotope or drug.

[0549] Technical Solution 204: A bispecific or multispecific molecule comprising a peptide as described in any one of Technical Solutions 1 to 175 and having binding specificity to human effector cells.

[0550] Technical Solution 205: A bispecific or multispecific molecule containing a peptide as described in any one of Technical Solutions 1 to 175 and having binding specificity to CD3, CD4, CD138, IL-15R, membrane-bound or receptor-bound TNF-α, human Fc receptor, or membrane-bound or receptor-bound IL-15.

[0551] Technical Solution 206: A pharmaceutical composition comprising a peptide as described in any one of Technical Solutions 1 to 176 or an immunoconjugate as described in Technical Solution 202 or 205 and a pharmaceutically acceptable carrier.

[0552] Technical Solution 207: The pharmaceutical composition described in Technical Solution 206 contains one or more other therapeutic agents.

[0553] Technical Solution 208: A method for inhibiting the growth and / or proliferation of cells expressing CD38, comprising administering a peptide as described in any one of Technical Solutions 1 to 176, an immunoconjugate as described in Technical Solutions 202 or 205, a pharmaceutical composition as described in Technical Solutions 206 or 207, or an expression vector as described in any one of Technical Solutions 178 to 182, thereby inhibiting the growth and / or proliferation of cells.

[0554] Technical Solution 209: A method for treating a disease or symptom in a patient involved in cells expressing CD38, the method comprising administering to a patient in need of treatment a peptide as described in any one of Technical Solutions 1 to 176, an immunoconjugate as described in Technical Solution 202 or 205, a pharmaceutical composition as described in Technical Solution 206 or 207, or an expression vector as described in any one of Technical Solutions 178 to 182.

[0555] Technical Solution 210: A method for inhibiting a disease or symptom in a patient caused by cells expressing CD38, the method comprising administering to a patient in need of treatment a peptide as described in any one of Technical Solutions 1 to 176, an immunoconjugate as described in Technical Solution 202 or 205, a pharmaceutical composition as described in Technical Solution 206 or 207, or an expression vector as described in any one of Technical Solutions 178 to 182.

[0556] Technical Solution 211: A method as described in Technical Solution 209 or 210, wherein the disease or symptom is rheumatoid arthritis.

[0557] Technical Solution 212: A method as described in Technical Solution 209 or 210, wherein the disease or symptom is multiple myeloma.

[0558] Technical Solution 213: A method as described in any one of Technical Solutions 209 to 212, wherein the method includes administering one or more other therapeutic agents to the patient.

[0559] Technical Solution 214: A method as described in Technical Solution 213, wherein the one or more other therapeutic agents are selected from chemotherapeutic agents, anti-inflammatory agents, or immunosuppressive and / or immunomodulatory agents.

[0560] Technical Solution 215: A method as described in Technical Solution 213, wherein the one or more other therapeutic agents are selected from platinum, gefitinib, cetuximab, rituximab, bevacizumab, erlotinib, bortezomib, thalidomide, pamidronate, zoledronic acid, clodronate, risedronate, ibandronate, etidronate, alendronate, tiludronate, arsenic trioxide, lenalidomide, filgrastim, pegylated filgrastim, sargramostim, suberoylanilide hydroxamic acid and SCIO-469.

[0561] Technical Solution 216: An in vitro method for detecting the presence of CD38 antigen or cells expressing CD38 in a sample, comprising: a) contacting the sample with a peptide as described in any one of Technical Solutions 1 to 176 under conditions where the peptide and CD38 can form a complex; and b) detecting the formation of the complex.

[0562] Technical Solution 217: An in vitro method as described in Technical Solution 216, wherein the peptide is an antibody.

[0563] Technical Solution 218: A kit for detecting the presence of CD38 antigen or cells expressing CD38 in a sample containing the peptide as described in any one of Technical Solutions 1 to 176.

[0564] Technical Solution 219: An in vivo method for detecting the presence of CD38 antigen or cells expressing CD38 in a patient, comprising: a) administering a peptide as described in any one of Technical Solutions 1 to 176 under conditions where the peptide and CD38 can form a complex; and b) detecting the formed complex.

[0565] Technical Solution 220: The in vivo method as described in Technical Solution 219, wherein the peptide is an antibody.

[0566] Technical solution 221: An anti-idiotypic antibody that binds to a peptide as described in any one of technical solutions 2, 4, or 163 to 174.

[0567] Technical Solution 222: Use of the anti-idiotypic antibody as described in Technical Solution 221 in detecting the level of a peptide as described in any one of Technical Solutions 2, 4 or 163 to 174 in a sample.

[0568] Technical Solution 223: Use of the anti-idiotypic antibody as described in Technical Solution 221 in detecting the level of human monoclonal antibodies against CD38 in a sample.

[0569] The terms "CD38" and "CD38 antigen" are used interchangeably herein and include any variant, isoform, and species homolog of human CD38 that is naturally expressed by cells or expressed in cells transfected with the CD38 gene. Synonyms for CD38 in the art include ADP-ribosyl cyclase 1, cADPr hydrolase 1, Cd38-rs1, cyclo-ADP-ribose hydrolase 1, 1-19, and NIM-R5 antigen.

[0570] The term peptide as used herein refers to both CD38-binding peptides and non-CD38 peptides, including any suitable peptide, and unless otherwise specified herein, can be used interchangeably with the terms polypeptide and protein; as long as the reader recognizes that each molecule comprising various amino acid polymers is significantly different and thus forms a single technical solution of the present invention (for example, a peptide comprising multiple polypeptide chains, such as an antibody, is significantly different from a single-chain antibody, a peptide immunoadhesive, or a single-chain immunopeptide). Therefore, the term peptide herein can be understood to refer to any suitable peptide of any suitable size and composition (including the number of amino acids in the protein molecule and the number of interacting chains).

[0571] In addition, unless otherwise indicated herein, the peptides of the inventive methods and compositions described herein may include non-naturally occurring and / or non-L amino acid residues. Unless otherwise indicated herein, the term peptide further (e.g., the term polypeptide and / or protein in the individual embodiments) also includes derivatized peptide molecules. In short, in the context of the present invention, a derivative is a peptide in which one or more amino acid residues in the peptide are chemically modified (e.g., alkylated, acylated, esterified or aminated), or connected to one or more non-amino acid organic and / or inorganic atoms or molecular substituents (e.g., polyethylene glycol (PEG) groups, lipophilic substituents (optionally, they can be connected to the amino acid sequence of the peptide via spacer residues or groups, such as β-alanine, γ-aminobutyric acid (GABA), UD-glutamic acid, succinic acid, etc.), fluorescent, biotin, radioactive nuclei, etc.), and may also or optionally contain non-essential, non-naturally occurring and / or non-L amino acid residues, unless otherwise specified in the text (but it should be recognized again that such derivatives themselves and the molecules they include can be regarded as independent features of the present invention, and the inclusion of such molecules in the concept of peptides is for the purpose of more convenient description of the present invention, rather than indicating any equivalence relationship between naked peptides and such derivatives). Non-limiting examples of such amino acid residues include, for example, 2-amino fatty acids, 3-amino fatty acids, β-alanine, β-aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminohexanoic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminocyclohexanoic acid, 2,4-diaminobutyric acid, desmosine, 2,2'-diaminocyclohexanoic acid, 2,3-diaminopropionic acid, N-ethylglycine, N-ethylaspartic acid, hydroxylysine, isohydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, isomeric isoleucine, N-methylglycine, 6-N-methyllysine, norvaline, norleucine, ornithine, and stedine-halogenated amino acids.

[0572] An antigen-binding peptide refers to any peptide that specifically binds to a portion of a given antigen under cellular and / or physiological conditions, thereby inducing, promoting, enhancing and / or regulating a physiological function associated with the given antigen for a sufficient period of time; it can be detected by ELISA, Western blotting or other similar suitable protein binding techniques described herein and / or known in the art and / or for a relevant period of time (e.g., at least about 15 minutes, at least about 30 minutes, at least about 45 minutes, at least about 1 hour, at least about 2 hours, at least about 4 hours, at least about 6 hours, at least about 12 hours, about 1-24 hours, about 1-36 hours, about 1-48 hours, about 1-72 hours, about 1 week or longer).

[0573] CD38 binding peptide, or CD38BP, is an antigen binding peptide that specifically binds to the antigen CD38. In one embodiment, the binding of CD38BP to CD38 is determined using the method described in Example 4.

[0574] The term immunoglobulin refers to a class of structurally related glycoproteins composed of two pairs of polypeptide chains, one pair of light (L) low molecular weight chains and one pair of heavy (H) chains, all four of which are interconnected by disulfide bonds. The structure of immunoglobulins has been described in detail. See, for example, Fundamental Immunology, Chapter 7 (Paul, W., ed., 2nd ed. Raven Press, NY (1989)). Briefly, each heavy chain typically includes a heavy chain variable region (abbreviated herein as V H ) and the heavy chain constant region. The heavy chain constant region typically includes three domains, C H 1. C H 2 and C H 3. Each light chain typically includes a light chain variable region (abbreviated herein as V L ) and a light chain constant region. The light chain constant region typically includes a domain C L . V H and V L The regions can be further subdivided into hypervariable regions (or structurally defined loops that can vary highly in sequence and / or form), also called complementarity determining regions (CDRs), interspersed with regions that are more conserved, called framework regions (FRs).

[0575] Each V H and V L Typically, it comprises three CDRs and four FRs, arranged from amino terminus to carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 (see also Chothia and Lesk J. MoI. Biol. 196, 901-917 (1987)). Typically, the number of amino acid residues in this region is determined by the method described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. (1991) (Phrases such as "variable domain residues numbered as in Kabat or according to Kabat herein" refer to the numbering system used for the heavy chain variable domain or the light chain variable domain). Using this numbering system, the actual linear amino acid sequence of a peptide may contain fewer or additional amino acids corresponding to a shortening or insertion of a FR or CDR of the variable domain. For example, the heavy chain variable domain may include at V HThere is a single amino acid insertion after residue 52 of CDR2 (residue 52a according to Kabat) and there are residue insertions after residue 82 of the heavy chain FR (e.g., residues 82a, 82b, and 82c according to Kabat). For a given antibody, the Kabat numbering of residues can be determined by aligning homologous regions of the antibody sequence with the "standard" Kabat numbering sequence.

[0576] The antibody (Ab) in the present invention refers to an immunoglobulin molecule, a portion of an immunoglobulin molecule or a derivative thereof, which has the ability to specifically bind to an antigen under typical physiological conditions for a sufficient time, such as at least about 30 minutes, at least about 45 minutes, at least about 1 hour, at least about 2 hours, at least about 4 hours, at least about 8 hours, at least about 12 hours, at least about 24 hours or more, at least about 48 hours or more, at least about 3, 4, 5, 6, 7 or more days, etc., or any other relevant functionally determined time period (e.g., sufficient to induce, promote, enhance and / or modulate a physiological response associated with antibody binding to an antigen).

[0577] The variable regions of the heavy and light chains of immunoglobulin molecules contain binding domains that interact with antigens. The constant regions of antibodies (Abs) mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classic complement system.

[0578] The anti-CD38 antibody can be a bispecific antibody, a bifunctional antibody, or a similar molecule (see, for example, the bifunctional antibody described in PNAS USA 90 (14), 6444-8 (1993)). In fact, the bispecific antibodies, bifunctional antibodies, etc. provided by the present invention can bind to any other suitable target besides the CD38 portion.

[0579] As mentioned above, unless otherwise indicated or clearly contradicted by the context, the term antibody herein includes portions of antibodies that have the ability to specifically bind to an antigen. It has been shown that the antigen binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments included in the term "antibody" include (i) Fab fragments, which contain V L 、V H 、C L and C H 1 domain; (ii) F(ab)2 and F(ab')2 fragments, bivalent fragments containing two Fab fragments connected by a disulfide bond in the hinge region; (iii) mainly composed of V H and C H (iv) an Fd fragment consisting mainly of a single arm of the antibody L and V HFv fragment composed of structural domains; (v) dAb fragment (Ward et al., Nature 341, 544-546 (1989)), which is mainly composed of V H domains; (vi) independent complementarity determining regions (CDRs) 1, and (vii) combinations of two or a single independent CDRs, optionally linked by a synthetic linker. L and V H The two domains are encoded by different genes, but they can be recombinantly synthesized by L and V H The present invention relates to a single-chain antibody or single-chain Fv (scFv) formed by connecting two or more domains to form a monovalent molecule (known as a single-chain antibody or single-chain Fv (scFv), see, for example, Bird et al., Science 242, 423-426 (1988) and Hustone et al., PNAS USA 85, 5879-5883 (1988)). Such single-chain antibodies are within the scope of the term antibody, unless otherwise specified or clearly indicated in the text. Other types of single-chain antibodies, such as bifunctional antibodies, are included within the scope of the term antibody. Although such fragments are generally included in the meaning of antibodies, they are all unique features of the present invention, exhibiting different biological properties and uses. These and other useful antibody fragments are further discussed herein.

[0580] It should also be understood that the term antibody also generally includes polyclonal antibodies, monoclonal antibodies (mAbs), antibody-like polypeptides, such as chimeric antibodies and humanized antibodies prepared by any known technology, such as enzymatic cleavage, peptide synthesis and recombinant technology, anti-idiotypic (anti-Id) antibodies of antibodies and antibody fragments with the ability to specifically bind to an antigen (antigen-binding fragments). The antibodies produced have any isotype.

[0581] The anti-CD38 antibody is an antibody as described above, which can specifically bind to the antigen CD38.

[0582] The term "antigenic determinant" refers to a protein-determining region that can specifically bind to an antibody. Antigenic determinants are usually composed of chemically active surface groups of molecules, such as amino acids or sugar side chains, and usually have specific three-dimensional structural features, as well as specific charge characteristics. Conformational and non-conformational antigenic determinants can be distinguished because the binding of the former is abolished in the presence of denaturing solvents, while the binding of the latter is not abolished. An antigenic determinant may contain amino acid residues that are directly involved in binding (also called the immunodominant component of the antigenic determinant) and other amino acid residues that are not directly involved in binding, such as amino acid residues that are effectively blocked by a specific antigen-binding peptide (i.e., amino acid residues in the footprint of a specific antigen-binding peptide).

[0583] The term "bispecific molecule" includes any agent, such as a protein, peptide, and a complex of a protein or peptide, that has two different binding specificities. For example, the molecule can bind to or interact with (a) an antigen on the surface of a cell and (b) an Fc receptor encoded by an effector cell. The term "multispecific molecule" includes any agent, such as a protein, peptide, and a complex of a protein or peptide, that has more than two different binding specificities. For example, the molecule can bind to or interact with (a) an antigen on the surface of a cell, (b) an Fc receptor encoded by an effector cell, and (c) at least one other component. Thus, the present invention includes, but is not limited to, bispecific, trispecific, tetraspecific, and other multispecific molecules directed against CD38 and other cell surface antigens or targets, such as Fc receptors of effector cells.

[0584] The term "bispecific antibody" includes any anti-CD38 antibody that is a bispecific molecule. The term "bispecific antibody" also includes bifunctional antibodies. Bifunctional antibodies are bivalent bispecific antibodies in which V H and V L The domains are expressed on a single polypeptide chain, but are connected by a short linker that prevents pairing between the two domains on the same chain, allowing the domains to pair with the complementary domains of another chain to create two antigen-binding sites (see, e.g., Holliger, P. et al., PNAS USA 90, 6444-6448 (1993); Poljak, RJ. et al., Structure 2, 1121-1123 (1994)).

[0585] As used herein, the term "effector cell" refers to an immune cell that participates in the effector phase of an immune response, corresponding to the cognitive and activation phase of an immune response. Exemplary immune cells include cells of bone marrow or lymphoid origin, for example, lymphocytes (e.g., B cells and T cells, including cytolytic T cells (CTLs)), killer cells, natural killer cells, macrophages, monocytes, eosinophils, neutrophils, segmented leukocytes, granulocytes, mast cells, and basophils. Certain effector cells express specific Fc receptors and have specific immune functions. In certain technical solutions, effector cells can induce antibody-dependent cytotoxicity (ADCC), for example, neutrophils can induce ADCC. For example, monocytes and macrophages expressing FcRs participate in specific target cell killing and present antigens to other components of the immune system, or bind to cells that present antigens. In certain technical solutions, effector cells can phagocytose target antigens, target cells, or microorganisms. The expression of specific FcRs on effector cells can be regulated by humoral factors, such as cytokines. For example, it has been found that FcγRI expression can be upregulated by interferon gamma (IFN-γ) and / or G-CSF. This enhanced expression can enhance the cytotoxic activity of FcγRI-containing cells against targets. Effector cells can phagocytose or lyse target antigens or target cells.

[0586] The term "human antibody" as used herein includes antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the present invention also include amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by in vivo cell mutation). However, the term "human antibody" as used herein does not include antibodies derived from other mammalian species, such as mice, whose CDR sequences are transplanted into human reading frame sequences. The human antibodies used herein are "derived from" specific germline sequences. If the antibodies are obtained from a system using human immunoglobulin sequences, such as by immunizing transgenic mice carrying human immunoglobulin genes, or by screening a human immunoglobulin gene library, the amino acid sequence of the screened human antibodies will be identical to that of the antibodies encoded by the germline V H or V L The amino acid sequence identity of the variable region gene segment is at least 90%, such as at least 95%, such as at least 96%, such as at least 97%, such as at least 98%, or such as at least 99%. Typically, the variable region gene segment is derived from a specific human germline V H or V L The variable region gene segment sequences of human antibodies have no more than 10 amino acid differences, such as no more than 5, for example no more than 4, 3, 2 or 1 amino acid differences, compared to the amino acid sequences encoded by the germline immunoglobulin genes.

[0587] Chimeric antibodies are antibodies that contain one or more regions derived from one antibody and one or more regions derived from one or more other antibodies from another species. A monovalent chimeric antibody is a dimer (HL) formed by connecting a chimeric H chain to a chimeric L chain via a disulfide bond. A bivalent chimeric antibody is a tetramer (H2L2) formed by two HL dimers connected by at least one disulfide bond. Multivalent chimeric antibodies can also be produced, for example, by oligomerizing the CH region (e.g., derived from an IgM H chain or μ chain). Typically, a chimeric antibody refers to an antibody whose heavy and / or light chain portions are identical or homologous to corresponding sequences of antibodies derived from a specific species or belong to a specific antibody type or subclass, while the remainder of the chain is identical or homologous to corresponding sequences of antibodies from another species or belongs to another antibody type or subclass, and fragments of such antibodies, as long as they have the desired biological activity (see, for example, US 4,816,567 and Morrison et al., PNAS USA 81., 6851-6855 (1984)). Chimeric antibodies are produced by recombinant procedures known in the art (see, e.g., Cabilly et al., PNAS USA 81, 3273-3277 (1984), Morrison et al., PNAS USA 8JL 6851-6855 (1984), Boulianne et al., Nature 312, 643-646 (1984), EP 125023, Neuberger et al., Nature 314, 268-270 (1985), EP 171496, EP 173494, WO 86 / 01533, EP 184187, Sahagan et al., J. Immunol. 137, 1066-1074 (1986), WO 87 / 02671, Liu et al., PNAS USA 84, 3439-3443 (1987), Sun et al., PNAS USA 84, 214-218 (1987), Better et al., Science 240, 1041-1043 (1988) and Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, (1988)).

[0588] Humanized antibodies are antibodies derived from non-human species in which specific amino acid mutations are made to the reading frame and the constant regions of the heavy and light chains to avoid or eliminate immune responses in humans. Humanized forms of non-human (e.g., murine) antibodies are chimeric antibodies containing minimal sequence derived from non-human immunoglobulins. For the most part, humanized antibodies are human immunoglobulins (recipient antibodies) in which the hypervariable region residues of the recipient are replaced by hypervariable region residues of a non-human species (donor antibody), such as mouse, rat, rabbit, or non-human primate, with the desired antigen-binding properties, such as specificity and affinity. In some cases, the Fv reading frame region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. In addition, humanized antibodies may contain residues that are not present in the recipient antibody or the donor antibody. These modifications can further optimize the function of the antibody. Typically, humanized antibodies contain at least one, typically two, variable domains in which all or substantially all of the loops in the hypervariable region correspond to non-human immunoglobulins, or all or substantially all of the FR regions are sequences of human immunoglobulins. The humanized antibody optionally also contains at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For details, see Jones et al., Nature 321, 522-525 (1986), Riechmann et al., Nature 332, 323-329 (1988), and Presta, Curr. Op. Struct. Biol. 2, 593-596 (1992).

[0589] As used herein, "monoclonal antibody" or "monoclonal antibody composition" refers to the preparation of an antibody molecule from a single molecular composition. A monoclonal antibody composition has a single binding specificity and affinity for a particular antigenic determinant. Thus, the term "human monoclonal antibody" refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences and having a single binding specificity. Human monoclonal antibodies can be produced by hybridomas, including B cells obtained from transgenic or transchromosomal non-human animals, such as transgenic mice, having a genome containing human heavy and light chain transgenes, and fused to immortalized cells. Monoclonal antibodies can be abbreviated as mAb.

[0590] The term "recombinant human antibody" as used herein includes all human antibodies prepared, expressed, generated or isolated by recombinant methods, such as (a) antibodies isolated from transgenic or transchromosomal animals (such as mice) containing human immunoglobulin genes or hybridomas prepared (described elsewhere herein), (b) antibodies isolated from host cells transformed to express antibodies, such as transfectomas, (c) antibodies isolated from recombinant, combinatorial human antibody libraries, and (d) antibodies prepared, expressed, generated or isolated by any other method involving splicing human immunoglobulin gene sequences with other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in some technical solutions, such recombinant human antibodies can be mutated in vitro (or in vivo somatic mutation when animal transgenes are used for human Ig sequences), so that the V H and V L The amino acid sequences of the regions, when they are derived from human germline V H and V L Sequences, when related thereto, may be sequences that are not naturally present in the germline repertoire of human antibodies in vivo.

[0591] As used herein, "heterologous antibody" is defined as an antibody produced by a transgenic non-human organism. The term refers to an antibody having an amino acid sequence corresponding to an antibody found in an organism other than a non-human animal, typically in a species other than the transgenic non-human animal.

[0592] As used herein, an "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds CD38 is substantially free of antibodies that specifically bind antigens other than CD38). However, an isolated antibody that specifically binds to an antigenic determinant, isoform, or variant of human CD38 may have cross-reactivity with other related antigens, such as those derived from other species (e.g., CD38 species homologs). Furthermore, the isolated antibody is substantially free of other cellular material and / or chemicals. In one embodiment of the present invention, a combination of "isolated" monoclonal antibodies having different specificities may comprise distinct components.

[0593] As used herein, "specific binding" refers to the binding of an antigen-binding peptide, such as an antibody, to a predetermined antigen. Typically, when measured by surface plasmon resonance (SPR) on a BIAcore 3000 instrument using recombinant CD38 as a ligand and an antibody as an analyte, the binding affinity of an antigen-binding peptide, such as an antibody, is equivalent to K D is 10 -7 M or less, for example about 10 -8 M or less, for example about 10 -9 M or less, about 10 -10 M or less, about 10-11 M or less. The antigen-binding peptide can bind to the predetermined antigen with a K D , binds with an affinity that is at least 10-fold lower, such as at least 100-fold lower, such as at least 1000-fold lower, such as at least 10,000-fold lower, such as at least 1,000,000-fold lower, than its binding affinity to a non-specific antigen other than the predetermined protein or a closely related antigen (e.g., BSA, casein). The amount of lower affinity depends on the K of the antigen-binding peptide. D , so when the K of the antigen-binding peptide D When the affinity is very low (i.e., the antigen-binding peptide is highly specific), the amount of affinity for the antigen is at least 10,000 times lower than the affinity for the non-specific antigen. As used herein, the phrases "antigen-binding peptide that recognizes an antigen" and "antigen-specific antigen-binding peptide" can be used interchangeably with the term "antigen-binding peptide that specifically binds to an antigen." Similarly, as used herein, the phrases "antibody that recognizes an antigen" and "antigen-specific antibody" can be used interchangeably with the term "antibody that specifically binds to an antigen."

[0594] The term "k d ”(sec -1 ) refers to the dissociation equilibrium rate constant for a specific antibody-antigen interaction. The value is also referred to as k off value.

[0595] The term "k a ”(M -1 ×sec -1 ) refers to the binding equilibrium rate constant for a specific antibody-antigen interaction.

[0596] The term "k D "(M) refers to the dissociation equilibrium constant of a specific antibody-antigen interaction.

[0597] The term "K A ”(M -1 ) refers to the binding equilibrium constant of a specific antibody-antigen interaction, expressed as k a Divide by k d And get.

[0598] As used herein, "isotype" refers to the antibody class (e.g., IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM) encoded by the heavy chain constant region genes.

[0599] As used herein, "isotype switching" refers to the phenomenon in which the class or isotype of an antibody changes from one immunoglobulin class to another immunoglobulin class.

[0600] As used herein, "non-switched isotype" refers to the isotype type of heavy chain produced when no isotype switching occurs; the CH gene encoding the non-switched isotype is typically the first CH gene immediately downstream of the functionally recombined VDJ gene. Isotype switching is categorized as either typical or atypical. Typical isotype switching occurs through a recombination event involving at least one switch sequence region of a transgene. Atypical isotype switching can occur, for example, by homologous recombination between human σμ and human ∑μ (δ-linked deletion). Alternative atypical switching mechanisms, such as intratransgenic and / or intrachromosomal recombination, can occur and achieve isotype switching.

[0601] As used herein, the term "switch sequence" refers to those DNA sequences responsible for switch recombination. The "switch donor" sequence is typically the μ switch region, located 5' (i.e., upstream) of the constant region that is deleted during switch recombination. The "switch acceptor" region is located between the deleted constant region and the replacement constant region (e.g., γ, ε, etc.). Because there is no specific site where recombination occurs, the final gene sequence is typically not predictable from the construct.

[0602] As used herein, "glycosylation pattern" is defined as the pattern of carbohydrate units covalently attached to a protein, more specifically an immunoglobulin (antibody) protein. The glycosylation pattern of the heterologous antibody is characterized by being substantially similar to the glycosylation pattern naturally occurring on antibodies produced by the non-human transgenic animal species. One of ordinary skill in the art will recognize that the glycosylation pattern of the heterologous antibody is more similar to the glycosylation pattern of the non-human transgenic animal species than to the species from which the transgenic CH gene is derived.

[0603] As used herein, the term "naturally occurring" when applied to a subject means that the subject is found in nature. For example, a polypeptide or polynucleotide sequence found in an organism (including viruses) that can be isolated from a source in nature and has not been artificially altered in the laboratory is naturally occurring.

[0604] As used herein, the term "rearranged" refers to a conformation of a heavy or light chain immunoglobulin locus in which the sequences encoding the complete V H or V L The domains are arranged in a configuration in which the V segments are positioned immediately adjacent to the DJ or J segments. Recombined immunoglobulin (antibody) gene loci can be identified by comparison to germline DNA; recombined loci have at least one heptamer / nonamer homology element.

[0605] As used herein, the term "unrearranged" or "germline conformation" with reference to a V segment refers to a conformation in which the V segment does not recombine to be in close proximity to a D or J segment.

[0606] As used herein, the term "nucleic acid molecule" refers to DNA molecules and RNA molecules. Nucleic acid molecules can be single-stranded or double-stranded, but are preferably double-stranded DNA. Nucleic acids can be present in whole cells, cell lysates, or in partially purified or substantially purified form.

[0607] A nucleic acid is "isolated" or "substantially purified" when it is purified from other cellular components or other contaminants such as other cellular nucleic acids or proteins by standard techniques, including alkaline / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other techniques well known in the art. See F. Ausubel et al., ed. Current Protocols in Molecular Biology, Greene Publishing and Wiley InterScience New York (1987).

[0608] A nucleic acid is "operably linked" when it is functionally linked to another nucleic acid sequence. For example, a promoter or enhancer is operably linked to a coding sequence when it affects the transcription of that sequence. For the transcription of a regulatory sequence, operably linked means that the linked DNA sequences are contiguous across the region connecting the two protein coding regions and that the reading frames are also continuous. For a switch sequence, operably linked means that the sequence can affect switch recombination.

[0609] As used herein, the term "inhibit growth" (e.g., when referring to cells) refers to any measurable decrease in cell growth when contacted with CD38BP, e.g., an anti-CD38 antibody, as compared to when not contacted with CD38BP, e.g., an anti-CD38 antibody, e.g., inhibition of cell culture growth by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99% or 100%.

[0610] As used herein, the terms "inhibit binding" and "block binding" (e.g., when referring to inhibition / blocking of binding of a CD38 binder to CD38) are used interchangeably and encompass both partial and complete inhibition / blocking. Inhibition / blocking of binding of a CD38 binder to CD38 can reduce or alter the normal level or type of cell signaling that occurs when binding of the CD38 binder to CD38 is not inhibited or blocked. Inhibition and blocking also refer to any measurable decrease in the binding affinity of a CD38 binder to CD38 when contacted with a CD38BP, such as an anti-CD38 antibody, compared to a ligand that has not been contacted with the CD38BP, such as an anti-CD38 antibody, e.g., inhibition of binding of a CD38 binder to CD38 by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100%.

[0611] "Target cell" refers to any undesired cell in a subject (e.g., a human or animal) that can be targeted by a composition of the invention (including, for example, a CD38BP such as a human monoclonal anti-CD38 antibody and / or a bispecific or multispecific molecule directed against CD38). In certain embodiments, a target cell is a cell that expresses or overexpresses CD38. Cells expressing CD38 typically include hematopoietic cells such as bone marrow thymocytes, activated T and B cells, the remaining 80% of NK cells and monocytes, lymphoblasts in lymph node germinal centers, plasma B cells and certain follicular cells, dendritic cells, normal bone marrow cells, specific precursor cells, 50-80% of umbilical cord blood cells, red blood cells, and platelets. CD38 is also expressed by non-hematopoietic cells such as epithelial cells and lamina propria lymphocytes in the gut, Purkinje cells and neurofibrillary tangles in the brain, epithelial cells of the prostate, beta cells in the pancreas, osteoclasts in bone, retinal cells in the eye, and sarcolemma in smooth and striated muscle. In malignant cells, CD38 is also expressed in various hematological malignancies, including but not limited to multiple myeloma, primary and secondary plasma cell leukemia, B-cell chronic lymphocytic leukemia, B-cell acute lymphoblastic leukemia, Waldenstrom macroglobulinemia, primary systemic amyloidosis, mantle cell lymphoma, prolymphocytic / myeloid leukemia, acute myeloid leukemia, chronic myeloid leukemia, follicular lymphoma, and NK cell leukemia.

[0612] As used herein, the term "vector" refers to a nucleic acid molecule that can transport another nucleic acid to which it is linked. One type of vector is a "plasmid," which refers to a circular double-stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, in which additional DNA segments can be ligated into the viral genome. Certain vectors can replicate autonomously in the host cell into which they are introduced (e.g., bacterial vectors with a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can integrate into the host cell's genome after introduction and then replicate along with the host genome. In addition, certain vectors can direct the expression of genes to which they are operably linked. Such vectors are referred to herein as "recombinant expression vectors" (or simply, "expression vectors"). In general, expression vectors used in recombinant DNA techniques are typically in the form of plasmids. In this specification, "plasmid" and "vector" are used interchangeably, as plasmids are the most commonly used form of vector. However, the present invention encompasses other forms of expression vectors, such as viral vectors (e.g., replication-defective retroviruses, adenoviruses, and adeno-associated viruses), which can perform the same function.

[0613] As used herein, the term "recombinant host cell" (or simply, "host cell") refers to a cell into which a recombinant expression vector has been introduced. It should be understood that the term refers not only to the specific subject cell, but also to the progeny of such a cell. Because certain changes may occur in progeny due to mutations or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term "host cell" as used herein. Recombinant host cells include, for example, transfectomas such as CHO cells, NS / 0 cells, and lymphocytes.

[0614] The term "regulatory sequence" includes promoters, enhancers, and other expression control elements (e.g., polyadenylation signals) that control the transcription or translation of the antibody chain genes. Such regulatory sequences can be described, for example, in Goeddel, Gene Expression Technology. Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990). One skilled in the art can design expression vectors, including selecting regulatory sequences based on factors such as the choice of host cell to be transformed and the level of expression of the desired protein. Examples of regulatory sequences for mammalian host cell expression include viral elements that direct high-level protein expression in mammals, such as promoters and / or enhancers derived from cytomegalovirus (CMV), simian virus 40 (SV40), adenovirus (e.g., adenovirus major late promoter (AdMLP)), and polyoma virus. Alternatively, non-viral regulatory sequences can be used, such as the ubiquitin promoter or the β-globin promoter.

[0615] The term "subject" as used herein includes any human and non-human animal. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0616] The term "transfection" in its various forms encompasses a wide variety of techniques commonly used to introduce exogenous DNA into prokaryotic or eukaryotic cells, such as electroporation, calcium phosphate precipitation, DEAE-dextran transfection, lipofection, and the like.

[0617] As used herein, the term "transfectoma" includes recombinant eukaryotic host cells that express the antibody, such as CHO cells, NS / 0 cells, HEK293 cells, plant cells, or fungi, including yeast cells.

[0618] The term "non-human animals" includes all vertebrates, for example, mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc. The term "non-human animals" includes all vertebrates, for example, mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc. The term "non-human animals" includes all vertebrates, for example, mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0619] The term "transgenic non-human animal" refers to a non-human animal that has a genome containing one or more human heavy and / or light chain transgenes or transchromosomes (integrated or not integrated into the animal's native genomic DNA) and is capable of fully expressing human antibodies. For example, a transgenic mouse may harbor a human light chain transgene or a human heavy chain transchromosome, thereby producing human CD38 antibodies when immunized with the CD38 antigen and / or cells expressing CD38. The human heavy chain transgene may be integrated into the mouse's chromosomal DNA, in which case the transgenic mouse is, for example, a HuMAb mouse such as the HCo7 or HCo12 mouse, or the human heavy chain transgene may be maintained extrachromosomally, in which case the transchromosomal KM mouse is described in WO 02 / 43478. Such transgenic and transchromosomal mice (collectively referred to herein as "transgenic mice") can produce a variety of human monoclonal antibody isotypes (e.g., IgG, IgA, IgM, IgD, and / or IgE) against a given antigen through VDJ recombination and isotype switching. Transgenic non-human animals can also be used to produce antibodies against specific antigens by introducing genes encoding specific antibodies, for example, by operatively linking the genes to genes expressed in the animal's milk.

[0620] As used herein, the term specificity refers to the ability of a CD38-binding peptide, such as an anti-CD38 antibody, to recognize a CD38 antigenic determinant while having minimal or no detectable reactivity toward other portions of CD38 (including other antigenic determinants that can be bound by other CD38BPs, such as anti-CD38 antibodies). Specificity can be measured using competition assays as described herein. Specificity can be more specifically measured using any of the antigenic determinant identification / delineation techniques described herein or equivalent techniques known in the art.

[0621] However, antibodies specific for particular antigenic determinants can cross-react with other molecules in certain biological samples where CD38 is present. More typically, CD38BPs, such as anti-CD38 antibodies, can cross-react with CD38 homologs from other species. In either or both of these cases, these cross-reactive antibodies are typically selected against structural and / or environmental factors associated with human CD38.

[0622] As used herein, the term "selective" refers to the preferential binding of a CD38BP, such as an anti-CD38 antibody, to a specific region, target, or peptide; typically, a region or antigenic determinant on CD38 relative to one or more other biomolecules, structures, cells, tissues, etc. In one embodiment, a CD38BP, such as an anti-CD38 antibody, of the present invention is selective for the CD38 portion of the environment of colorectal cancer cells (i.e., the anti-CD38 antibody selectively binds to the CD38 portion over other components of colorectal cancer cells).

[0623] The CD38BPs of the present invention are typically used and provided in an at least substantially isolated form. A substantially isolated molecule is one that is the predominant species in a composition comprising the molecule and its class of molecules (i.e., it comprises at least about 50% of the molecular species in the composition, and typically comprises at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or more of the molecular species, e.g., peptides, in the composition (e.g., the composition is at least about 98%, 98%, or 99% homogeneous with respect to all peptide species present)).

[0624] An isolated molecule is one that is not significantly (e.g., greater than about 1%, greater than about 2%, greater than about 3%, or greater than about 5%) associated with any exogenous and non-essential physiological factors, such as non-CD38 binding biomolecules present in the cell or animal producing CD38BP (or CD38 binding molecules that may interfere with CD38BP binding and / or activity of the present invention). An isolated molecule may also refer to any molecule obtained through a purification stage involving human intervention (whether automated, manual, or both). In the various compositions provided herein, for example, compositions containing one or more pharmaceutically acceptable carriers (e.g., compositions containing a large amount of pharmaceutically acceptable carriers, stabilizers, and / or preservatives), CD38BP may be present in relatively small amounts relative to the total number of molecular species in the composition. In some cases, additional peptides, such as BSA, may also be included in such compositions along with previously purified CD38BP. However, if the additional composition of such a composition is acceptable for the intended use of CD38BP, such a composition may still be described as comprising isolated CD38BP.

[0625] Typically, the CD38BPs of the present invention are substantially free of other CD38BPs, such as CD38BPs with different antigenic specificities. However, the present invention also provides compositions containing multiple CD38BPs with different specificities and characteristics (e.g., the present invention provides CD38BPs "cocktails" with different specificities and / or selectivity characteristics).

[0626] "Treatment" refers to the administration of an effective dose of a therapeutically active compound of the present invention to achieve the purpose of alleviating, reducing or eradicating (curing) symptoms or disease states.

[0627] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 2. L CD38BP.

[0628] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 6. H CD38BP.

[0629] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 2. L and V consisting essentially of the sequence of SEQ ID No: 6 H CD38BP.

[0630] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 3. L CD38BP of CDR1.

[0631] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 4. L CD38BP of CDR2.

[0632] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 5. L CD38BP of CDR3.

[0633] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 8. H CD38BP of CDR1.

[0634] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 9. H CD38BP of CDR2.

[0635] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 10. H CD38BP of CDR3.

[0636] In one embodiment, the present invention provides a V comprising a sequence consisting essentially of SEQ ID No: 3, SEQ ID No: 4 and SEQ ID No: 5.L CDRs(V L CDR1, CDR2 and CDR3) of CD38BP.

[0637] In one embodiment, the present invention provides a V comprising a sequence consisting essentially of SEQ ID No: 8, SEQ ID No: 9 and SEQ ID No: 10. H CDRs(V H CDR1, CDR2 and CDR3) of CD38BP.

[0638] In one technical solution, the present invention provides CD38BP, which comprises

[0639] (a) Three types of V L The CDRs consist essentially of SEQ ID No: 3, SEQ ID No: 4, and SEQ ID No: 5, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). L spacers between CDRs), and

[0640] (b) Three types of V H The CDRs consist essentially of SEQ ID No: 8, SEQ ID No: 9, and SEQ ID No: 10, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). H between the CDRs).

[0641] In a further technical solution, the present invention provides a V L Area and V H In another further technical solution, the present invention provides a CD38BP, wherein V L and V H The V domain is located on different chains of the immunoglobulin fold protein and is L CDR1, CDR2, CDR3 and V H CDR1, CDR2 and CDR3 are sequentially and directionally connected, thereby selectively and / or specifically binding to the antigenic determinant on CD38. In another further technical solution, the present invention provides a sequence comprising two sets of variable domains (V L and V H domain group), so that CD38BP contains two identical antigen determinant binding sites.

[0642] It is contemplated that any such CD38BPs described in this paragraph, at least in part, may be conjugated to a V having a sequence comprising SEQ ID No: 2. L Region and V containing SEQ ID No: 7 sequence H Antibodies targeting the same region have similar antigenic determinant specificity, selectivity and other characteristics and can therefore be used to treat multiple myeloma.

[0643] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 12. L CD38BP.

[0644] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 17. H CD38BP.

[0645] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 12. L and V consisting essentially of the sequence of SEQ ID No: 17 H CD38BP.

[0646] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 13. L CD38BP of CDR1.

[0647] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 14. L CD38BP of CDR2.

[0648] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 15. L CD38BP of CDR3.

[0649] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 18. H CD38BP of CDR1.

[0650] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 19. H CD38BP of CDR2.

[0651] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 20. H CD38BP of CDR3.

[0652] In one embodiment, the present invention provides a V comprising a sequence consisting essentially of SEQ ID No: 13, SEQ ID No: 14 and SEQ ID No: 15. L CDRs(V L CDR1, CDR2 and CDR3) of CD38BP.

[0653] In one embodiment, the present invention provides a V comprising a sequence consisting essentially of SEQ ID No: 18, SEQ ID No: 19 and SEQ ID No: 20. H CDRs(V H CDR1, CDR2 and CDR3) of CD38BP.

[0654] In one technical solution, the present invention provides CD38BP, which comprises

[0655] (a) Three types of V L The CDRs consist essentially of SEQ ID No: 13, SEQ ID No: 14, and SEQ ID No: 15, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). L spacers between CDRs), and

[0656] (b) Three types of V H The CDRs consist essentially of SEQ ID No: 18, SEQ ID No: 19, and SEQ ID No: 20, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). H between the CDRs).

[0657] In a further technical solution, the present invention provides a V L Area and V H In another further technical solution, the present invention provides a CD38BP, wherein V L and V H The V domain is located on different chains of the immunoglobulin fold protein and is L CDR1, CDR2, CDR3 and V H CDR1, CDR2 and CDR3 are sequentially and directionally connected, thereby selectively and / or specifically binding to the antigenic determinant on CD38. In another further technical solution, the present invention provides a sequence comprising two sets of variable domains (V L and V Hdomain group), so that CD38BP contains two identical antigen determinant binding sites.

[0658] It is contemplated that any such CD38BPs described in this paragraph, at least in part, may be conjugated to a V comprising the sequence of SEQ ID No: 12. L Region and V containing SEQ ID No: 17 sequence H Antibodies targeting the same region have similar antigenic determinant specificity, selectivity and other characteristics and can therefore be used to treat multiple myeloma.

[0659] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 22. L CD38BP.

[0660] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 27. H CD38BP.

[0661] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 22. L and V consisting essentially of the sequence of SEQ ID No: 27 H CD38BP.

[0662] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 23. L CD38BP of CDR1.

[0663] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 24. L CD38BP of CDR2.

[0664] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 25. L CD38BP of CDR3.

[0665] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 28. H CD38BP of CDR1.

[0666] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 29. H CD38BP of CDR2.

[0667] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 30. H CD38BP of CDR3.

[0668] In one embodiment, the present invention provides a V comprising a sequence consisting essentially of SEQ ID No: 23, SEQ ID No: 24 and SEQ ID No: 25. L CDRs(V L CDR1, CDR2 and CDR3) of CD38BP.

[0669] In one embodiment, the present invention provides a V comprising a sequence consisting essentially of SEQ ID No: 28, SEQ ID No: 29 and SEQ ID No: 30. H CDRs(V H CDR1, CDR2 and CDR3) of CD38BP.

[0670] In one technical solution, the present invention provides CD38BP, which comprises

[0671] (a) Three types of V L The CDRs consist essentially of SEQ ID No: 23, SEQ ID No: 24, and SEQ ID No: 25, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). L spacers between CDRs), and

[0672] (b) Three types of V H The CDRs consist essentially of SEQ ID No: 28, SEQ ID No: 29, and SEQ ID No: 30, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). H between the CDRs).

[0673] In a further technical solution, the present invention provides a V L Area and V H In another further technical solution, the present invention provides a CD38BP, wherein V L and V H The V domain is located on different chains of the immunoglobulin fold protein and is L CDR1, CDR2, CDR3 and V HCDR1, CDR2 and CDR3 are sequentially and directionally connected, thereby selectively and / or specifically binding to the antigenic determinant on CD38. In another further technical solution, the present invention provides a sequence comprising two sets of variable domains (V L and V H domain group), so that CD38BP contains two identical antigen determinant binding sites.

[0674] It is contemplated that any such CD38BPs described in this paragraph, at least in part, may be conjugated to a V comprising the sequence of SEQ ID No: 22. L Region and V containing SEQ ID No: 27 sequence H Antibodies targeting the same region have similar antigenic determinant specificity, selectivity and other characteristics and can therefore be used to treat multiple myeloma.

[0675] In one embodiment, the present invention provides a V sequence consisting essentially of SEQ ID No: 3 or SEQ ID No: 13 or SEQ ID No: 23. L CD38BP of CDR1, wherein the N-terminal residue and / or 1, 2 or 3 C-terminal amino acid residues are deleted.

[0676] In one embodiment, the present invention provides a V sequence consisting essentially of SEQ ID No: 4 or SEQ ID No: 14 or SEQ ID No: 24. L CD38BP of CDR2, wherein the N-terminal residue and / or 1, 2 or 3 C-terminal residues are deleted.

[0677] In one embodiment, the present invention provides a V sequence consisting essentially of SEQ ID No: 5 or SEQ ID No: 15 or SEQ ID No: 25. L CD38BP of CDR3, wherein the N-terminal residue and / or 1, 2, 3 or 4 C-terminal residues are deleted.

[0678] In one embodiment, the present invention provides a V sequence consisting essentially of SEQ ID No: 8 or SEQ ID No: 18 or SEQ ID No: 28. H CD38BP of CDR1, wherein 1, 2, 3 or 4 N-terminal residues and / or 1, 2, 3 or 4 C-terminal residues are deleted.

[0679] In one embodiment, the present invention provides a V sequence consisting essentially of SEQ ID No: 9 or SEQ ID No: 19 or SEQ ID No: 29. H CD38BP of CDR2, wherein 1, 2, 3, 4 or 5 N-terminal amino acids and / or 1, 2, 3, 4, 5 or 6 C-terminal amino acids are deleted.

[0680] In one embodiment, the present invention provides a V comprising a sequence substantially consisting of SEQ ID No: 10 or SEQ ID No: 20 or SEQ ID No: 30. H CD38BP of CDR3, wherein the N-terminal 1, 2 or 3 amino acid residues and / or the C-terminal 1, 2, 3 or 4 amino acid residues are deleted.

[0681] The present invention also provides CD38BPs in which these "spliced" CDR sequences are combined with each other and / or other CDR sequences described herein.

[0682] In one technical solution, the present invention provides CD38BP, comprising

[0683] (a) Three types of V L The CDRs consist essentially of SEQ ID No: 3, SEQ ID No: 4, and SEQ ID No: 5, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). L spacers between CDRs), and

[0684] (b) Three types of V H The CDRs consist essentially of SEQ ID No: 8, SEQ ID No: 9, and SEQ ID No: 10, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). H between the CDRs).

[0685] In a further technical solution, the present invention provides a V L Area and V H In another further technical solution, the present invention provides a CD38BP, wherein V L and V H The V domain is located on different chains of the immunoglobulin fold protein and is L CDR1, CDR2, CDR3 and V HCDR1, CDR2 and CDR3 are sequentially and directionally connected, thereby selectively and / or specifically binding to the antigenic determinant on CD38. In another further technical solution, the present invention provides a sequence comprising two sets of variable domains (V L and V H It is expected that any such CD38BPs described in this paragraph will, at least in part, be conjugated to a V domain comprising SEQ ID No: 2. L Region and V containing SEQ ID No: 7 sequence H Antibodies targeting the same region have similar antigenic determinant specificity, selectivity and other characteristics.

[0686] In one technical solution, the present invention provides CD38BP, comprising

[0687] (a) Three types of V L The CDRs consist essentially of SEQ ID No: 13, SEQ ID No: 14, and SEQ ID No: 15, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). L spacers between CDRs), and

[0688] (b) Three types of V H The CDRs consist essentially of SEQ ID No: 18, SEQ ID No: 19, and SEQ ID No: 20, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). H between the CDRs).

[0689] In a further technical solution, the present invention provides a V L Area and V H CD38BP contains a flexible linker between the domains.

[0690] In another further technical solution, the present invention provides a CD38BP, wherein V L and V H The V domain is located on different chains of the immunoglobulin fold protein and is L CDR1, CDR2, CDR3 and V H CDR1, CDR2 and CDR3 are sequentially and directionally connected, thereby selectively and / or specifically binding to the antigenic determinant on CD38. In another further technical solution, the present invention provides a sequence comprising two sets of variable domains (V Land V H It is expected that any of the CD38BPs described in this paragraph will, at least in part, be conjugated to a V domain comprising SEQ ID No: 12. L Region and V containing SEQ ID No: 17 sequence H Antibodies targeting the same region have similar antigenic determinant specificity, selectivity and other characteristics.

[0691] In one technical solution, the present invention provides CD38BP, comprising

[0692] (a) Three types of V L The CDRs consist essentially of SEQ ID No: 23, SEQ ID No: 24, and SEQ ID No: 25, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). L spacers between CDRs), and

[0693] (b) Three types of V H The CDRs consist essentially of SEQ ID No: 28, SEQ ID No: 29, and SEQ ID No: 30, respectively, and are located adjacent to each other in CD38BP (e.g., near V in wild-type anti-CD38 antibody). H between the CDRs).

[0694] In a further technical solution, the present invention provides a V L Area and V H CD38BP contains a flexible linker between the domains.

[0695] In another further technical solution, the present invention provides a CD38BP, wherein V L and V H The V domain is located on different chains of the immunoglobulin fold protein and is L CDR1, CDR2, CDR3 and V H CDR1, CDR2 and CDR3 are sequentially and directionally connected, thereby selectively and / or specifically binding to the antigenic determinant on CD38. In another further technical solution, the present invention provides a sequence comprising two sets of variable domains (V L and V H It is expected that any such CD38BPs described in this paragraph will, at least in part, be conjugated to a V domain comprising the sequence of SEQ ID No: 22.L Region and V containing SEQ ID No: 27 sequence H Antibodies targeting the same region have similar antigenic determinant specificity, selectivity and other characteristics.

[0696] The present invention also provides V comprising the antibody of the embodiment L Area V H CD38BPs with functional variants of the CDRs or one or more CDRs. L 、V H Functional variants of the CDRs or CDRs still allow CD38BP to retain at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the affinity / potency and / or specificity / selectivity of the parent antibody, and in some cases, such CD38BP can have higher affinity, selectivity and / or specificity than the parent antibody.

[0697] In one technical solution, the present invention provides a method comprising a variable V L CD38BP, the V L Substantially consisting of a sequence having about 50%, such as at least 60%, for example at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least 95% amino acid sequence identity to the sequence of SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of the antibody, the antibody having a variable V of SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, respectively. L Sequences, such as V having SEQ ID No: 2 L Sequence and SEQ ID No: 7 V H The antibody of sequence, and V respectively having SEQ ID No: 12 L Sequence and SEQ ID No: 17 V H The antibody of sequence, and V respectively having SEQ ID No: 22 L Sequence and SEQ ID No: 27 V H Sequence of antibodies.

[0698] In one technical solution, the present invention provides a method comprising a variable V L CDR1 of CD38BP, the V LThe CDR1 consists essentially of a sequence having about 50%, such as at least 60%, for example at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity to the sequence set forth in SEQ ID No: 3 or SEQ ID No: 13 or SEQ ID No: 23, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of an antibody having a variable V Fc of SEQ ID No: 3 or SEQ ID No: 13 or SEQ ID No: 23, respectively. L CDR1 sequence, such as an antibody having SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, such as an antibody having SEQ ID No: 2 or SEQ ID No: 22, respectively. L Sequence and SEQ ID No: 7 V H The antibody of sequence, and V respectively having SEQ ID No: 12 L Sequence and SEQ ID No: 17 V H and V having SEQ ID No: 22, respectively. L Sequence and SEQ ID No: 27 V H Sequence of antibodies.

[0699] In one technical solution, the present invention provides a method comprising a variable V L CDR2 of CD38BP, the V L The CDR2 consists essentially of a sequence having about 50%, such as at least 60%, for example at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity to the sequence set forth in SEQ ID No: 4 or SEQ ID No: 14 or SEQ ID No: 24, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of an antibody having a variable V Fc of SEQ ID No: 4 or SEQ ID No: 14 or SEQ ID No: 24, respectively. L CDR2 sequence, such as an antibody having SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, such as an antibody having SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, respectively. L Sequence and SEQ ID No: 7 V H The antibody of sequence, and V respectively having SEQ ID No: 12L Sequence and SEQ ID No: 17 V H and V having SEQ ID No: 22, respectively. L Sequence and SEQ ID No: 27 V H Sequence of antibodies.

[0700] In one technical solution, the present invention provides a method comprising a variable V L CDR3 of CD38BP, the V L The CDR3 consists essentially of a sequence having about 50%, such as at least 60%, for example at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity to the sequence set forth in SEQ ID No: 5 or SEQ ID No: 15 or SEQ ID No: 25, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of an antibody having a variable V L CDR3 sequences, such as antibodies having SEQ ID No: 2 or SEQ ID No: 12 or SEQ ID No: 22, respectively, such as antibodies having V sequences of SEQ ID No: 2, respectively. L Sequence and SEQ ID No: 7 V H The antibody of sequence, and V respectively having SEQ ID No: 12 L Sequence and SEQ ID No: 17 V H and V having SEQ ID No: 22, respectively. L Sequence and SEQ ID No: 27 V H Sequence of antibodies.

[0701] In one technical solution, the present invention provides a method comprising a variable V H CD38BP, the V HSubstantially consisting of a sequence having about 50%, such as at least 60%, for example at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity to the sequence set forth in SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of the antibody having a variable V of SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, respectively. L CDR3 sequences, such as V and V sequences having SEQ ID No: 7, H Sequence and SEQ ID No: 2 V L The antibody of sequence, and V respectively having SEQ ID No: 17 H Sequence and SEQ ID No: 12 V L and V having SEQ ID No: 27, respectively. H Sequence and SEQ ID No: 22 V L Sequence of antibodies.

[0702] In one technical solution, the present invention provides a method comprising a variable V H CDR1 of CD38BP, the V H The CDR1 consists essentially of a sequence having about 50%, such as at least 60%, for example at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity to the sequence set forth in SEQ ID No: 8 or SEQ ID No: 18 or SEQ ID No: 28, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of an antibody having a variable V Fc of SEQ ID No: 8 or SEQ ID No: 18 or SEQ ID No: 28, respectively. H CDR1 sequence, such as an antibody having SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, such as an antibody having SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, respectively. H Sequence and SEQ ID No: 2 V L and V sequences of SEQ ID No: 17. H Sequence and SEQ ID No: 12 V Land V having SEQ ID No: 27, respectively. H Sequence and SEQ ID No: 22 V L Sequence of antibodies.

[0703] In one technical solution, the present invention provides a method comprising a variable V H CDR2 of CD38BP, the V H The CDR2 consists essentially of a sequence having about 50%, such as at least 60%, for example at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity to the sequence set forth in SEQ ID No: 9 or SEQ ID No: 19 or SEQ ID No: 29, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of an antibody having a variable V Fc of SEQ ID No: 9 or SEQ ID No: 19 or SEQ ID No: 29, respectively. H CDR2 sequence, such as an antibody having SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, such as an antibody having SEQ ID No: 7 or SEQ ID No: 17 or SEQ ID No: 27, respectively. H Sequence and SEQ ID No: 2 V L and V sequences of SEQ ID No: 17. H Sequence and SEQ ID No: 12 V L and V having SEQ ID No: 27, respectively. H Sequence and SEQ ID No: 22 V L Sequence of antibodies.

[0704] In one technical solution, the present invention provides a method comprising a variable V H CDR3 of CD38BP, the V HThe CDR3 consists essentially of a sequence having about 50%, such as at least 60%, for example at least about 70%, such as at least about 75%, such as at least about 80%, such as at least about 85%, such as at least about 90%, such as at least about 95% amino acid sequence identity to the sequence set forth in SEQ ID No: 10 or SEQ ID No: 20 or SEQ ID No: 30, wherein CD38BP has at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the antigenic determinant binding characteristics of an antibody having a variable V Fc of SEQ ID No: 10 or SEQ ID No: 20 or SEQ ID No: 30, respectively. H CDR3 sequences, such as antibodies having SEQ ID No: 7, SEQ ID No: 17, or SEQ ID No: 27, respectively, such as antibodies having V sequences of SEQ ID No: 7, respectively. H Sequence and SEQ ID No: 2 V L The antibody of sequence, and V respectively having SEQ ID No: 17 H Sequence and SEQ ID No: 12 V L and V having SEQ ID No: 27, respectively. H Sequence and SEQ ID No: 22 V L Sequence of antibodies.

[0705] The percent identity between two sequences is a function of the number of identical sites in the sequences (i.e., % homology = # identical sites / total # sites × 100). For optimal alignment of the two sequences, the number of gaps and the length of each gap need to be considered. A mathematical algorithm can be used to compare the sequences between two sequences and determine the percent identity, as described in the following non-limiting examples.

[0706] The percent identity between two nucleotide sequences can be determined using the GCG software package (available from http: / / www.gcg.comThe percent identity between two nucleotide or amino acid sequences can also be determined using the GAP program in (available from) using the NWSgapdna.CMP matrix and a gap weight of 40, 50, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. The percent identity between two nucleotide or amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller, Comput. Appl. Biosci 4, 11-17 (1988), which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Alternatively, the percent identity between two amino acid sequences can be determined using the algorithm of Needleman and Wunsch, J. MoI. Biol. 48, 444-453 (1970), which has been incorporated into the GCG software package (available from http: / / www.gcg.com The GAP program in (obtained) was used with the Blossum 62 matrix and the PAM250 matrix, gap weights of 16, 14, 12, 10, 8, 6, or 4, and length weights of 1, 2, 3, 4, 5, or 6.

[0707] The nucleic acid and protein sequences of the present invention can further be used as a "query sequence" to search public databases, for example to identify related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul et al., J. MoI. Biol. 215, 403-10 (1990). BLAST nucleotide searches can be performed using the NBLAST program with a score of 100 and a word length of 12 to obtain nucleotide sequences homologous to the nucleic acid molecules of the present invention. BLAST protein searches can be performed using the XBLAST program with a score of 50 and a word length of 3 to obtain amino acid sequences homologous to the protein molecules of the present invention. To obtain gapped alignments for comparison, Gapped BLAST can be used as described in Altschul et al., Nucleic Acids Res. 25 (17), 3389-3402 (1997). When using BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See http: / / www.ncbi.nlm.nih.gov .

[0708] The sequence of a CDR variant can differ from the CDR sequence of the parent antibody sequence by a majority of conservative substitutions; for example, at least about 35%, about 50% or more, about 60% or more, about 70% or more, about 75% or more, about 80% or more, about 85% or more, about 90% or more, about 95% or more (e.g., about 65-99%) of the substitutions in the variant are replacements of conservative amino acid residues. In the context of the present invention, conservative substitutions are defined as substitutions in the amino acid classes set forth in one or more of the following three tables:

[0709] Types of conservatively substituted amino acid residues

[0710] acidic residues Asp and Glu basic residues Lys, Arg, and His Hydrophilic uncharged residues Ser, Thr, Asn, and Gln Aliphatic uncharged residues Gly, Ala, Val, Leu, and Ile Nonpolar uncharged residues Cys, Met, and Pro Aromatic residues Phe, Tyr, and Trp

[0711] Types of conservative amino acid residue substitutions that can be selected

[0712] 1 Ala(A) Ser(S) Thr(T) 2 Asp(D) Glu(E) 3 Asp(N) Gln(Q) 4 Arg(R) Lys(K) 5 Ile(l) Leu(L) Met(M) 6 Phe(F) Tyr(Y) Trp(W)

[0713] Physical and functional classification of alternative amino acid residues

[0714] Residues containing hydroxyl groups S and T Aliphatic residues I, L, V, and M Residues connected to cycloolefin groups F, H, W, and Y hydrophobic residues A, C, F, G, H, I, L, M, R, T, V, W, and Y Negatively charged residues D and E Polar residues C, D, E, H, K, N, Q, R, S, and T Positively charged residues H, K, and R Small residues A, C, D, G, N, P, S, T, and V Very small residues A, G, and S Residues involved in turn formation A, C, D, E, G, H, K, N, Q, R, S, P, and T Flexible residues Q, T, K, S, G, P, D, E, and R

[0715] More conservative substitution groups include: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine and aspartic acid-glutamic acid. Other groups of amino acids can also be represented by the principles described in Creighton (1984) Proteins: Structure and Molecular Properties (2d Ed. 1993), WH Freeman and Company.

[0716] In one embodiment of the present invention, compared to the CDRs of the antibodies in the examples, the conservation of hydrophilic / hydrophobic properties and residue molecular weight / size in the variant CDRs is substantially retained (e.g., the molecular weight class, hydropathic score, or both of the sequence are retained by at least about 50%, at least about 60%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95% or more (e.g., 65-99%)). For example, substitution of conserved residues can also be selectively based on the strength or weakness of the molecular weight of the conserved group, as is known in the art.

[0717] Alternatively, the conservation of similar residues can also be determined by similarity scores, such as by using a BLAST program (e.g., BLAST 2.2.8 available through NCBI). Typically, suitable variants have at least about 45%, such as at least about 55%, at least about 65%, at least about 75%, at least about 85%, at least about 90%, at least about 95% or more (e.g., about 70-99%) similarity to the parent peptide.

[0718] Substantial changes in function can be produced by selecting substitutions that are less conservative than those in the above-defined groups. For example, non-conservative substitutions can be made that more significantly affect the structure of the peptide in the region of the change, such as an α-helix or a β-sheet; the charge or hydrophobicity of the molecule at the target site, or steric hindrance of the side chain. Substitutions generally expected to produce the greatest changes in the properties of the polypeptide are those where 1) a hydrophilic residue, such as seryl or threonyl, is replaced by (or is replaced by) a hydrophobic residue, such as leucyl, isoleucyl, phenylalanyl, valyl, or alanyl; 2) a cysteine ​​or proline residue is replaced by (or is replaced by) any other residue; 3) a residue with a positively charged side chain, such as lysyl, arginyl, or histidyl, is replaced by (or is replaced by) a negatively charged residue, such as glutamyl or aspartyl; or 4) a residue with a sterically hindered side chain, such as phenylalanine, is replaced by (or is replaced by) a residue without a side chain, such as glycine. Thus, these and other non-conservative substitutions may be introduced into peptide variants where significant changes in function / structure are desired and such changes that eliminate conservation of structure / function are desirable.

[0719] A simple way to generate alternative variants is to use affinity maturation techniques using phage using methods known in the art. In order to identify candidate hypervariable region sites for modification, alanine scanning mutagenesis can be performed to identify hypervariable regions that contribute significantly to antigen binding. Optionally or additionally, it is useful to analyze the crystal structure of the antigen-antibody complex to identify contact sites between the antigen and antibody. These contact residues and adjacent residues may be suitable candidates for substitution.

[0720] When creating variant antibodies by inserting hypervariable regions, the typical range of hypervariable region lengths studied in known antibodies should be considered. For example, for the first hypervariable region of the light chain variable domain, the length of the parent antibody V can be increased while maintaining substantial similarity and the expected appropriate size. L Insertions are introduced into CDR1, typically V according to Kabat et al., supra. L CDR1 has a total of about 9-20 (e.g., about 10-17) residues. L CDR2 has a total length of about 5-10 residues; typically V LCDR3 is approximately 7-20 residues long; typically V H CDR1 is approximately 10-15 residues long; typically V H CDR2 is approximately 15-20 residues long; typically V H CDR3 is about 6-30 residues long (e.g., 3-25 residues). Using the alignment and counting method described in Kabat, typically, H CDR3 generates a V H Typically, the insertion is near the C-terminus of the domain, e.g., in the parent V H CDR3 at about residues 97-102 (e.g., adjacent to, or C-terminally adjacent to, the parent V H (e.g., residue 100 of the CDR3 sequence). Antibody variants having inserted amino acid residues in their hypervariable regions can be randomly generated, particularly where the initial binding affinity of the parent antibody for the target antigen allows for easy screening of the randomly generated antibody variants. For example, phage display provides a method for screening such random variants.

[0721] When designing, constructing and / or evaluating CDR variants, it should be noted that the CDR region can be changed to better bind to the antigenic determinant. Typically, antibody CDRs can be manipulated by providing a complementary surface, which may include a finger-like structure that can extend to the surface of the antigenic protein or other antibody determinant structures that fix the antigenic determinant. If the antigenic determinant is not tightly fixed, the antibody will not exert maximum affinity. But for the antigenic determinant, in the antibody determinant structure, it is usually several key residues that are mainly responsible for this binding. Therefore, CDR sequences can have significant differences in length and composition between antibodies against the same peptide. Technicians know that residues such as tyrosine residues (for example, at V H Specific residues in the CDR3 sequence that typically contribute significantly to binding to this antigenic determinant are typically retained in the CDR variants.

[0722] Variants in the CDR regions may also increase the amino acid contacts between the antigen and the antibody variant by introducing one or more amino acid residues (by substitution or insertion) that increase the contacts or energetically favorable interactions between one or more amino acid residues present in the antigen and one or more amino acid residues present in the antibody, compared to the amino acid contacts between the antigen and the parent antibody. The amino acid interactions of interest may be selected from hydrophobic interactions, van der Waals interactions, and ionic interactions.

[0723] Those skilled in the art will be aware of additional principles in designing and screening CD38BP variants containing the CDRs of the antibodies of the present invention.

[0724] For CDR variants of the antibodies in the embodiments, particularly variant CDRs in anti-CD38 antibodies or fragments thereof, typically, residues required for supporting and / or orienting the CDR structural loop structure are retained; typically, residues located within about 10 angstroms of the CDR structural loop (but selectively only residues in the region having a water solvent contact surface of about 5 square angstroms or greater) are not altered or are altered only by conservative amino acid residue substitutions; and / or typically, the amino acid sequence has only a limited number of insertions and / or deletions (if any) so that the CDR structural loop-like structure is retained in the variant (for a description of the relevant techniques and related principles, see, for example, Schiweck et al., J MoI Biol. 268(5), 934-51 (1997); Morea, Biophys Chem. 68(1-3), 9-16 (1997); Shirai et al., FEBS Lett. 399(1-2), 1-8 (1996); Shirai et al., FEBS Lett. 455(1-2), 188-97 (1999), Reckzo et al., Protein Eng. 8(4), 389-95 (1995) and Eigenbrot et al., J MoI Biol. 229(4), 969-95 (1993). See also WO 03 / 048185, WO 03 / 070747 and WO 03 / 027246.

[0725] Other techniques can also be used to generate variant antibodies, including directed evolution and other variant generation techniques, for example, in US20040009498, Marks et al., Methods MoI Biol. 248, 327-43 (2004), Azriei-Rosenfeld et al., J MoI Biol. 335(1), 177-92 (2004), Park et al., Biochem Biophys Res Commun. 275(2). 553-7 (2000), Kang et al., Proc Natl Acad Sci US A. 88(24), 11120-3 (1991), Zahnd et al., J Biol Chem. 279(18), 18870-7 (2004), Xu et al., ChemBiol. 9(8), 933-42 (2002), Border et al., Proc As described in Natl Acad Sci US A. 97(20), 10701-5 (2000), Crameri et al., Nat Med. 2(1), 100-2 (1996), and more generally, for example, in WO 03 / 048185.

[0726] The resulting antibody variants can be used in any suitable screening technique, and antibodies that possess suitable and desired superior characteristics in one or more relevant assays can be screened for further investigation.

[0727] The CD38BPs containing CDR sequences as described above retain at least a substantial portion (at least about 50%, 60%, 70%, 80%, 90%, 95% or more) of the V sequences as in SEQ ID No: 2. L Sequence and V in SEQ ID No: 7 H Antibodies having sequences, and / or having V sequences as in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H Antibodies having sequences, and / or having V sequences as in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 H In the case of antibodies with different affinity / strength and / or specificity / selectivity, any suitable number and combination of such V sequences may be contained. L and V HCDRs, but may optionally differ in other properties such as immunogenicity in human patients, affinity for antigenic determinants, increased half-life, etc. In some cases, such CD38BPs may have higher affinity, selectivity and / or specificity than the parent antibody. In one embodiment, less than one complete set of V sequences may be present in the CD38BP of the present invention. L CDRs and / or V H CDRs. In one embodiment, all V L CDRs and V H CDRs.

[0728] Examples of other functional properties of antibodies that may be altered or retained in the variant CD38BPs of the present invention compared to -003, -005, and -024 include:

[0729] (1) High affinity binding to CD38;

[0730] (2) low dissociation rate from CD38;

[0731] (3) inhibiting or blocking the binding of CD38 to its target;

[0732] (4) elimination of CD38-expressing T cells or B cells;

[0733] (5) induce high levels of CDC in CD55 / 59-negative or CD55 / 59-positive cells;

[0734] (6) translocation to lipid rafts after binding to CD38;

[0735] (7) T cell tolerance;

[0736] (8) inhibiting the proliferation of T or B cells expressing CD38;

[0737] (9) CD38 internalization;

[0738] (10) inhibit or induce the enzymatic activity of CD38;

[0739] (11) inhibit or induce CD38-induced signal transduction;

[0740] (12) Induce or inhibit the production of cytokines;

[0741] (13) Induce or block the differentiation of T cells or B cells;

[0742] (14) Induction or recovery from apoptosis;

[0743] (15) Reduce or increase the lytic inducibility of NK cells;

[0744] (16) Induce or inhibit insulin production in β cells in the pancreas;

[0745] (17) prolonging the survival of patients with tumor cells expressing CD38; and / or

[0746] (18) When mixed with an appropriate amount of effector cells, they induce ADCC of CD38 targets. The present invention also provides CD38BPs, characterized in that they can be combined with V having SEQ ID No: 2 L Sequence and V in SEQ ID No: 7 H An antibody having a sequence (such as antibody-003), or having a V sequence as in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H An antibody having a sequence (such as antibody-005), or having a V sequence as in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 H Antibodies to the same sequence (such as Antibody-024) compete (competitively inhibit) or cross-compete (ie, relatively partially inhibit binding of an antigenic determinant) for binding to CD38.

[0747] For example, the CD38BP may be a Fab fragment derived from an antibody that binds to a protein having V as in SEQ ID No: 2. L Sequence and V in SEQ ID No: 7 H Antibodies having sequences, and / or having V sequences as in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H Antibodies having sequences, and / or having V sequences as in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 H The Fab fragments are small compared to mAb molecules and therefore cannot significantly compete with the antibodies for binding to CD38, even though the antibodies from which they are derived can do so. However, these CD38BPs also play a role in similar localization near the CD38 region (e.g., localization of cytotoxins, radionucleosides, etc., within the CD38BP of immunoconjugates). Therefore, these CD38BPs can be used in the methods of the present invention and are therefore provided herein.

[0748] Competition between two or more CD38BPs for binding to CD38 or a portion of CD38 can be determined by any suitable technique. In one embodiment, for example, competition is determined as described in Examples 7, 8, and 9.

[0749] Competition in the context of the present invention refers to any detectable, significant reduction in the binding affinity between a particular molecule and a particular binding partner in the presence of another molecule that binds to the binding partner. Typically, competition refers to a reduction of at least about 10%, such as at least about 15%, or at least about 20%, in the binding of CD38BP to another CD38BP, as determined by, for example, an ELISA assay or FACS assay (as described in the Examples) using sufficient amounts of two or more competing CD38BPs and CD38BPs molecules.

[0750] (a) CD38 form (e.g., “processed,” “mature,” “unprocessed,” “unprocessed,” or “immature” CD38);

[0751] (b) free CD38 forms (e.g., CD38 fragments generated by in vivo processing);

[0752] (c) a heterologous dipeptide consisting of another peptide related to CD38, such as CD31, and CD38;

[0753] (d) a complex consisting of CD38 and one or more substrates such as cAMP, NAD+ and / or cADPR;

[0754] (e) dimerized, associated and / or processed dimers of CD38 with a soluble ligand such as CD31; or

[0755] (f) Portions of CD38. For more than one CD38 and / or CD38 portion, for example, where the antibody binding properties of a specific region of CD38 are retained in the fragment, competition also occurs between CD38BPs. In this case, the fully presented linear antigenic determinant or conformational antigenic determinant located on multiple test fragments is presented on a sufficiently large CD38 fragment, as in CD38.

[0756] Typically, testing for competition involves assessing relative inhibition of binding using a first amount of a first molecule; a second amount of a second molecule; and a third amount of a third molecule (or a standard determined by binding studies that is more reasonably comparable to new binding data using the first and second molecules as surrogates for actual concurrent data), where the first, second, and third amounts are sufficient to allow comparison and thus provide information about the selectivity and / or specificity of the molecule in question relative to other available molecules. The first, second, and third amounts may vary depending on the nature of CD38BP and the potential target in question. For example, for an ELISA assay, similar to that described in the Examples section, approximately 5-50 μg (e.g., approximately 10-50 μg, approximately 20-50 μg, approximately 5-20 μg, approximately 10-20 μg, etc.) of CD38BP and / or CD38 target is required to test for competition. Conditions should also be suitable for binding. Typically, physiological or near-physiological conditions (e.g., temperature of approximately 20-40°C, pH of approximately 7-8, etc.) are suitable for CD38BP:CD38 binding.

[0757] Typically, competition is characterized by a relative inhibition significantly greater than about 5% as determined by ELISA and / or FACS analysis. A higher threshold for relative inhibition may be desirable as a standard / measure of an appropriate level of competition in a particular context (e.g., when the competition assay is used to select or screen for new antibodies designed to block the binding function of another peptide or molecule that binds to CD38 (e.g., a natural binding partner of CD38, such as CD31, also known as the CD31 antigen, EndoCAM, GPIIA1, PECAM-1, a platelet / endothelial cell adhesion molecule, or a naturally occurring anti-CD38 antibody). Thus, for example, a competition standard may be set whereby a relative inhibition of at least about 10% may be detected, a relative inhibition of at least about 15% may be detected, or a relative inhibition of at least about 20% may be detected before an antibody is considered sufficiently competitive. In cases where the antigenic determinant belonging to the competing antibody is in close proximity to the antigen, competition can be greater than 40% relative inhibition of CD38 binding (e.g., at least about 45% inhibition, such as about 50% inhibition, for example at least about 55% inhibition, such as about 60% inhibition, such as at least about 65% inhibition, such as about 70% inhibition, such as at least about 75% inhibition, such as about 80% inhibition, such as at least about 85% inhibition, such as about 90% inhibition, such as at least about 95% inhibition or higher levels of relative inhibition).

[0758] Competition can be considered the other side of cross-reactivity between a molecule and two potential binding partners. In certain embodiments, the CD38BP of the present invention specifically binds to one or more residues or regions of CD38, but does not cross-react with other peptides, peptide regions, or molecules. For example, the present invention provides an anti-CD38 antibody that does not cross-react with proteins homologous to CD38, such as BST-1 (bone marrow stromal cell antigen-1) and Mo5, also known as CD157; or an anti-CD38 antibody that does not cross-react with CD38 in normal tissues, such as tissues involved in multiple myeloma. Typically, lack of cross-reactivity means that the relative competitive inhibition between the molecules is less than 5% when assessed by ELISA and / or FACS analysis using sufficient amounts of the molecules under appropriate detection conditions.

[0759] In one embodiment, the present invention provides a method for combining with an antibody such as antibody-003 having a V sequence as in SEQ ID No: 2. L Sequence and V in SEQ ID No: 7 H Antibodies to the sequences compete for binding to CD38 or a portion thereof, CD38BP.

[0760] In one embodiment, the present invention provides a method for combining with an antibody such as antibody-005 having a V sequence as in SEQ ID No: 12. L Sequence and V in SEQ ID No: 17 H Antibodies to the sequences compete for binding to CD38 or a portion thereof, CD38BP.

[0761] In one embodiment, the present invention provides a method for combining with an antibody such as antibody-024 having a V sequence as in SEQ ID No: 22. L Sequence and V in SEQ ID No: 27 H Antibodies to the sequences compete for binding to CD38 or a portion thereof, CD38BP.

[0762] As described elsewhere herein, unless otherwise specified or clearly contradicted by the context, references to CD38BP binding to CD38 refer to binding in any suitable context, such as in the context of a conformational CD38 structure or in the context of a linear antigenic determinant. Clearly, binding in a limited subset of such contexts is an important property of any CD38BP provided herein.

[0763] Other methods for determining CD38BP specificity by competitive inhibition can be found, for example, in Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988), Colligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc, and Wiley InterScience NY, (1992, 1993), and Muller, Meth. Enzymol. 92, 589-601 (1983)).

[0764] Human CD38 contains multiple different antigenic determinants, including (1) peptide antigenic determinants contained within a single peptide chain of human CD38; (2) conformational antigenic determinants consisting of one or more non-contiguous amino acids on a particular chain and / or amino acids that are spatially contiguous but on different peptide chains (typically, where the respective amino acid sequences of the chains are non-contiguous in the human CD38 polypeptide sequence); (3) post-translational antigenic determinants consisting of molecular structures such as carbohydrate groups that are covalently attached in whole or in part to human CD38; or (4) combinations of (1)-(3).

[0765] Antigenic determinants in the present invention include any peptide or peptide-derived determinant that can specifically bind to an immunoglobulin. Antigenic determinants can comprise any suitable number or combination of amino acids (thus, at least partially charged) in any position (with respect to the linear sequence of CD38) or orientation (with respect to folded CD38 or fragments thereof).

[0766] Thus, for example, an antigenic determinant may comprise about 3-10 amino acids, typically 3-8 amino acids, at one or more consecutive or discontinuous positions in the primary sequence of CD38 (e.g., an antigenic determinant may consist essentially of 2, 3, 4, 5, 6, 7, or 8 amino acid residues distributed at 1, 2, 3, 4, or 5 discontinuous positions in CD38). Alternatively, for example, an antigenic determinant may be considered to be defined by a region of about 5-40 consecutive amino acid residues (e.g., about 7-30 amino acid residues, about 5-20 amino acid residues, or about 3-15 amino acid residues) on CD38 (either alone or in combination with a portion of an adjacent CD38 domain). In some antigenic determinants, only one or a few amino acid residues may be important for recognition by a CDR or CDR(s) (and therefore, most important for CD38BP:CD38 antigen affinity and strength). Thus, an antigenic determinant may be described based on one or more such key residues, while recognizing that other residues may also have smaller contributions to the antigenic determinant. In cases where the antigenic determinant is defined by an amino acid region, one or more amino acids in the region may contribute little or nothing to antibody binding, and thus such residues may be replaced with a suitable different residue without resulting in "loss" of the antigenic determinant for at least some CD38BPs specific for it.

[0767] In one embodiment, the present invention provides a CD38BP such as an anti-CD38 antibody, which can specifically bind to a CD38 antigenic determinant, which can also be represented by a V sequence as shown in SEQ ID No: 2. L Sequence and V in SEQ ID No: 7 H Antibodies with sequences such as Antibody-003, or antibodies with V sequences such as those in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H Antibodies with sequences such as Antibody-005, or antibodies with V sequences such as those in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 H The antibody (such as antibody-024) specifically binds to the sequence. L Sequence and V in SEQ ID No: 7 H CDRs of an antibody having the sequence of SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H CDRs of an antibody having the sequence of SEQ ID No: 22, or V L Sequence and V in SEQ ID No: 27H The CD38BPs of the CDRs of the antibody with the sequence of SEQ ID No: 2 can be specifically directed against the CDRs of the antibody with the sequence of SEQ ID No: 2. L Sequence and V in SEQ ID No: 7 H An antibody having a sequence as in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H An antibody having a sequence as in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 H In some cases, the CD38BP can recognize or be compared to a V sequence as in SEQ ID No: 2. L Sequence and V in SEQ ID No: 7 H An antibody having a sequence as in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H An antibody having a sequence as in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 H Sequence-specific antibodies are more specific / selective against a particular structure or region of the antigenic determinant.

[0768] Combined with V as in SEQ ID No: 2 L Sequence and V in SEQ ID No: 7 H Antibodies with sequences such as Antibody-003, or antibodies with V sequences such as those in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H Antibodies with sequences such as Antibody-005, or antibodies with V sequences such as those in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 HThe CD38 antigenic determinants of a sequenced antibody (e.g., Antibody-024) can be identified using standard mapping and characterization techniques. Further details can be identified using suitable techniques, and various examples of such techniques are available to the skilled artisan. These techniques can also be used to identify and / or characterize antigenic determinants against CD38BPs. In embodiments of such mapping / characterization methods, antigenic determinants for anti-CD38 antibodies can be determined by chemically modifying exposed amino / carboxyl groups in the CD38 protein using a "footprinting" of the antigenic determinant. A specific example of such a footprinting technique is the use of HXMS (hydrogen-deuterium exchange with mass spectrometry detection), in which hydrogen / deuterium exchange, binding, and back-exchange of amino protons of the receptor and ligand proteins occur, with backbone amino groups involved in protein binding protected during the back-exchange and therefore remaining deuterated. Relevant regions are then identified by peptide fragmentation, rapid-mixing high-performance liquid chromatography separation, and / or electrospray ionization mass spectrometry. See, for example, Ehring H, Analytical Biochemistry, 267 (2) 252-259 (1999) and / or Engen, JR and Smith, DL (2001) Anal. Chem. 73, 256A-265A. Another example of a suitable antigenic determinant identification technique is nuclear magnetic resonance antigenic determinant mapping (NMR), wherein typically, the positions of signals in a two-dimensional NMR spectrum of a free antigen and an antigen in complex with an antigen-binding peptide such as an antibody are compared. Typically, antigens are selectively detected using isotopes. 15 N labeling is used so that only signals associated with the antigen are visible on the NMR spectrum, while no signals are seen for the antigen bound to the peptide. Typically, compared to the spectrum of the free antigen, the antigen signals derived from amino acids and involved in the antigen-peptide interaction are shifted in the spectrum of the complex, allowing the amino acids involved in the binding to be identified. See, for example, Ernst Schering Res Found Workshop. (44), 149-67 (2004), Huang et al., Journal of Molecular Biology 281 (1), 61-67 (1998) and Saito and Patterson, Methods. 9 (3), 516-24 (1996).

[0769] Mass spectrometry can also be used for antigenic determinant mapping / delineation. See, for example, Downward, J Mass Spectrom. 35(4), 493-503 (2000) and Kiselar and Downard, Anal Chem. 71.(9), 1792-801 (1999).

[0770] Protease digestion technology can also be used for antigenic determinant mapping and identification. The region / sequence associated with the antigenic determinant can be determined by protease digestion, for example, by digestion with trypsin at a ratio of about 1:50 to CD38 at 37°C, pH 7-8, overnight (O / N), followed by mass spectrometry (MS) analysis for peptide identification. Peptides protected by CD38BP during trypsin cleavage can then be identified by comparing the trypsin-digested sample with an incubation with CD38BP followed by, for example, trypsin digestion (thereby indicating the footprint of the binding partner). Other enzymes such as chymosin, pepsin, etc. can also or alternatively be used as the same antigenic determinant description method. In these assays, peptides with V as in SEQ ID No: 2 are identified. L Sequence and V in SEQ ID No: 7 H Antibodies with sequences such as Antibody-003, or antibodies with V sequences such as those in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H Antibodies with sequences such as Antibody-005, or antibodies with V sequences such as those in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 H The CD38BP having the same antibody sequence as that in SEQ ID No: 2 (such as antibody-024) can be considered to be the same as that having V in SEQ ID No: 2. L Sequence and V in SEQ ID No: 7 H Antibodies with sequences such as Antibody-003, or antibodies with V sequences such as those in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H Antibodies with sequences such as Antibody-005, or antibodies with V sequences such as those in SEQ ID No: 22 L Sequence and V in SEQ ID No: 27 H Antibodies with the same sequence (e.g., Antibody-024) bind to the same antigenic determinant. For example, see Manca, Ann 1st Super Sanita. 27(1), 15-9 (1991) for a discussion of similar techniques. Another method for identifying regions of related antigenic determinants is to map the antigenic determinants obtained by competing for binding of two antibodies, one of which is biotinylated, to CD38.

[0771] Another method for identifying relevant antigenic determinant regions is to detect antibody binding to linear or cyclic CD38 peptides using a PEPSCAN-based enzyme-linked immunosorbent assay, see, for example, Slootstra-JW et al. Mol-Divers. 1, 87-96 (1996). Site-directed mutagenesis is another method for identifying relevant antigenic determinant regions, see, for example, Polyak and Deans, Blood 99, 3956-3962 (2002).

[0772] Various phage display technologies can also be used to identify antigenic determinants. See, for example, Wang and Yu, Curr Drug Targets. 5(1), 1-15 (2004), Burton, Immunotechnology. 1(2), 87-94 (1995 Aug), Cortese et al., Immunotechnology. 1(2), 87-94 (1995) and Irving et al., Curr Opin Chem Biol. 5(3), 314-24 (2001). Consistent antigenic determinants can also be identified by modified phage display-related technologies (see http: / / www.cs.montana.edu / ~mumey / papers / jcb03.pdf ).

[0773] Other methods for mapping antigenic determinants include crystallographic techniques, X-ray diffraction techniques (such as the X-ray diffraction / sequence analysis technique developed by Poljak and others in the 1970s-1980s), and the use of polynucleotide synthesis. Computer-based methods such as sequence analysis and three-dimensional structural analysis can also be used to identify antigenic determinants. For example, antigenic determinants can be determined by molecular modeling of the structure of CD38 and the structure of the individual monoclonal antibody Fab fragments that were entered. These and other mapping methods are discussed in Epitope Mapping A Practical Approach (Westwood and Hay Eds.) 2001 Oxford University Press.

[0774] In one technical solution, the present invention provides a method of producing a compound having one or more V selected from the group consisting of V in SEQ ID No: 2. L Sequence and V in SEQ ID No: 7 H Antibodies with sequences such as Antibody-003, or antibodies with V sequences such as those in SEQ ID No: 12 L Sequence and V in SEQ ID No: 17 H Antibodies with sequences such as Antibody-005, or antibodies with V sequences such as those in SEQ ID No: 22 LSequence and V in SEQ ID No: 27 H The mAbs of the same sequence (such as antibody-024) have the same specific CD38 binding characteristics as CD38BP.

[0775] Mapping studies have shown that several monoclonal antibodies that enhance human CD38 bind to an antigenic determinant (220-296) in the C-terminal region of CD38 (Hoshino et al. and Ferrero et al.). Three amino acid differences exist between the human and macaque CD38 sequences in this region: T237, Q272, and S274 in humans correspond to A238, R273, and F275 in macaques. -005 does not bind to macaque tissue (as shown in Examples 10 and 11). A limited number of amino acid differences exist between human and macaque CD38 sequences, for example, in the carboxyl-terminal portion of the protein. For example, the following three amino acid differences between human and macaque CD38 sequences: T237, Q272, and S274 in human CD38s correspond to A238, R273, and F275 in macaque CD38s (compare SEQ ID No. 21 and SEQ ID No. 22). The extent to which -005 binds to a mutant human CD38 protein in which the glutamine residue at position 272 of SEQ ID No: 31 is replaced with an arginine residue (Q272R), or a mutant human CD38 protein in which the serine residue at position 274 of SEQ ID No: 31 is replaced with a phenylalanine residue (S274F) (as described in Example 17), differs from its binding to wild-type human CD38. Specifically, binding to -005 was abolished by the amino acid substitution at position S274F.

[0776] Therefore, the present invention provides a peptide that binds to human CD38 (SEQ ID No: 31), and that binds to a mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is substituted with an arginine residue to a degree different from that of its binding to human CD38 (SEQ ID No: 31).

[0777] The present invention also provides a peptide that binds to human CD38 (SEQ ID No: 31), and the degree of binding to mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is substituted with a phenylalanine residue is different from the degree of binding to human CD38 (SEQ ID No: 31).

[0778] The term "to the same extent" should be understood as meaning that the binding of the peptide to the mutant human CD38 is significantly less than the binding of the peptide to wild-type human CD38. Binding of the peptide to CD38 molecules (wild-type and mutant) can be determined by various methods well known to those skilled in the art to determine whether binding to the mutant is "significantly less" than binding to the wild-type. A variety of techniques for determining binding of a peptide to another peptide can be used by those skilled in the art, such as ELISA, radioimmunoassay, BIAcore, or flow cytometry.

[0779] One method of determining binding is by measuring the EC of peptide binding to the mutant protein. 50 , and then comparing the values ​​obtained. Another method of measuring binding is by measuring the amount of binding at saturating concentrations (e.g., a plateau in the binding signal), or by measuring the kinetic rate constant k by, for example, BIAcore. on and k off .

[0780] In one embodiment, the binding of the peptide to CD38 protein (mutant or wild type) is determined by ELISA as described in Example 17.

[0781] In one embodiment, the peptide binds to an EC in which the serine residue at position 274 is replaced by a phenylalanine residue and a mutant human CD38 (SEQ ID No: 34) is present. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 In one embodiment, the peptide binds to an EC of a mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 In one embodiment, the peptide binds to an EC of a mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 In one embodiment, the peptide binds to an EC of a mutant human CD38 (SEQ ID No: 34) in which the serine residue at position 274 is replaced by a phenylalanine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 1%.

[0782] In one embodiment, the peptide binds to an EC in which the glutamine residue at position 272 is replaced by an arginine residue and the mutant human CD38 (SEQ ID No: 33) is substituted.50 The peptide binds to human CD38 (SEQ ID No: 31) 50 In one embodiment, the peptide binds to an EC of a mutant human CD38 (SEQ ID No: 33) in which the glutamine residue at position 272 is replaced by an arginine residue. 50 The peptide binds to human CD38 (SEQ ID No: 31) 50 10% of.

[0783] In one embodiment, the peptide binds to a mutant human CD38 (SEQ ID No: 32) in which the threonine residue at position 237 is replaced by an alanine residue, to the same extent as it binds to human CD38 (SEQ ID No: 31). In one embodiment, the peptide binds to a mutant human CD38 (SEQ ID No: 32) in which the threonine residue at position 237 is replaced by an alanine residue. 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 In one embodiment, the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 In one embodiment, the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 In one embodiment, the peptide binds to a mutant human CD38 in which the threonine residue at position 237 is replaced by an alanine residue (SEQ ID No: 32). 50 Greater than the EC of peptide binding to human CD38 (SEQ ID No: 31) 50 95% of.

[0784] To identify more specific potential antigenic determinant regions in CD38, various predictive analytical methods can be used. In the first analytical method, CD38 can be analyzed for (1) highly hydrophobic regions (using the Kyte-Doolittle method); (2) antigenicity as determined by the proto-convex area index method; (3) antigenicity as determined by the Parker method; (4) antigenicity as determined by the Hopp / Woods method; and (5) hydrophilicity as determined by the Goldman, Engleman, and Steitz method. Sequences of 10-40 amino acids in length can be selected based on their ability to possess one or more of these properties. The principle underlying this method is the general consensus that many ideal B cell antigenic determinants are hydrophilic, surface-oriented, and flexible sequences of 8-10 amino acids in length.

[0785] The present invention provides CD38BPs targeting the CD38 regions of CD38 identified in this manner. Furthermore, the termini of these sequences can be compared with regions predicted to contain antigenic determinants obtained through other analyses described herein, thereby providing additional specific regions that may contain antigenic determinants. Other similar comparisons can be performed to provide additional possible antigenic determinant regions, wherein CD38BPs that bind to these antigenic determinant regions can be considered another feature of the present invention.

[0786] In one embodiment, the CD38BP of the present invention is an antibody. Non-limiting examples of CD38-binding immunoglobulin molecules provided by the present invention include (a) a fully functional immunoglobulin molecule comprising: (i) two identical chimeric heavy chains comprising a variable region specific for a human B cell surface antigen and a human constant region, and (ii) two identical fully (i.e., non-chimeric) human light chains; (b) a fully functional immunoglobulin molecule comprising: (i) two chimeric heavy chains comprising the variable region and human constant region as described above, and (ii) two identical fully (i.e., non-chimeric) non-human light chains; and (c) a monovalent antibody, i.e., a fully functional immunoglobulin molecule comprising: (i) two chimeric heavy chains comprising the variable region and human constant region as described above, and (ii) two different light chains, only one of which has the same specificity as the heavy chain variable region. The resulting antibody molecule binds only to one end and is incapable of bivalent binding. As an example, the immunoglobulin-related peptides provided by the present invention may include the following components: (a) a whole immunoglobulin molecule; (b) scFv; (c) a monoclonal antibody; (d) a human antibody; (e) a chimeric antibody; (f) a humanized antibody; (g) a Fab fragment; (h) a Fab1 fragment; (i) a F(ab')2 fragment; (j) an Fv molecule; and (k) a disulfide-bonded Fv molecule.

[0787] In one embodiment, the CD38BP of the present invention is a polyclonal antibody. In one embodiment, the CD38BP of the present invention is a monoclonal antibody. In another embodiment, the CD38BP of the present invention is a human monoclonal antibody. In another embodiment, the CD38BP of the present invention is a humanized antibody. In another embodiment, the CD38BP of the present invention is a chimeric antibody. In another embodiment, the CD38BP of the present invention is a monoclonal antibody derived entirely from a mammalian species other than humans. In another embodiment, the CD38BP of the present invention is a completely murine monoclonal antibody.

[0788] A monoclonal antibody is a component of a homologous group of antibodies with identical structure and specificity. Typically, a monoclonal antibody is an antibody obtained from substantially homologous antibodies, i.e., a single antibody comprising identical group composition except for the presence of minor naturally occurring mutations. Monoclonal antibodies are highly specific, with each monoclonal antibody typically directed against a single antigenic determinant. In contrast, polyclonal antibody preparations typically include different antibodies directed against different antigenic determinants. The fact that an antibody is monoclonal does not necessarily mean that the antibody must be produced by any particular method. For example, the monoclonal antibodies of the present invention can be produced by the hybridization method described in Kohler et al., Nature 256, 495 (1975), or can be produced by recombinant DNA methods. Monoclonal antibodies can also be isolated from phage antibody libraries using techniques such as those described in Clackson et al., Nature 352, 624-628 (1991) and Marks et al., J. Mol. Biol. 222, 581-597 (1991).

[0789] Monoclonal antibodies can be obtained from any suitable source. Thus, for example, monoclonal antibodies can be obtained from hybridomas prepared from splenic B cells of mice immunized with the antigen of interest, e.g., in the form of cells expressing the antigen on their surface, or in the form of a nucleic acid encoding the antigen of interest. Monoclonal antibodies can also be obtained from hybridomas derived from antibody-expressing cells of immunized humans or non-human mammals such as mice, dogs, primates, etc.

[0790] Alternatively, cloned antibody genes can be expressed in other expression systems, including prokaryotes, such as microorganisms such as E. coli, algae, and insect cells, for the production of single-chain Fv antibodies. In addition, antibodies can be produced in transgenic non-human animals such as goat and rabbit milk or eggs, or in transgenic plants. For example, Verma, R., et al., J. Immunol. Meth. 216, 165-181 (1998); Pollock, et al., J. Immunol. Meth. 231, 147-157 (1999); and Fischer, R., et al., Biol. Chem. 380, 825-839 (1999).

[0791] In one embodiment, transgenic or transchromosomal mice carrying portions of the human immune system, rather than the murine system, can be used to generate human monoclonal antibodies directed against CD38. Such transgenic and transchromosomal mice include HuMAb mice and KM mice, respectively, referred to herein, and are collectively referred to herein as "transgenic mice." Human monoclonal antibodies generated in such mice are referred to as HuMabs.

[0792] HuMAb mice contain human immunoglobulins encoding unrearranged human heavy chain (μ and γ) and κ light chain immunoglobulin sequences with targeted mutations inactivating the mini-locus and the endogenous μ and κ chain gene loci (Lonberg, N. et al., Nature 368, 856-859 (1994)). As a result, the mice exhibit reduced expression of mouse IgM or κ during immune response, and the introduced human heavy and light chain transgenes undergo class switching and somatic mutation to generate high-affinity human IgG, κ monoclonal antibodies (Lonberg, N. et al. (1994), supra; reviewed in Lonberg, N. Handbook of Experimental Pharmacology 113, 49-101 (1994), Lonberg, N. and Huszar, D., Intern. Rev. Immunol. Vol. 1365-93 (1995) and Harding, F. and Lonberg, N. Ann. NY Acad. Sci 764536-546 (1995)). The preparation of HuMAb mice is described in detail in Taylor, L. et al., Nucleic Acids Research 20, 6287-6295 (1992), Chen, J. et al., International Immunology 5, 647-656 (1993), Tuaillon et al., J. Immunol. 152, 2912-2920 (1994), Taylor, L. et al., International Immunology 6, 579-591 (1994), and Fishwild, D. et al., Nature Biotechnology 14, 845-851 (1996). See also, US 5,545,806, US 5,569,825, US 5,625,126, US 5,633,425, US 5,789,650, US 5,877,397, US 5,661,016, US 5,814,318, US 5,874,299, US 5,770,429, US 5,545,807, WO 98 / 24884, WO94 / 25585, WO 93 / 1227, WO 92 / 22645, WO 92 / 03918 and WO 01 / 09187.

[0793] The HCo7 mouse has a JKD disruption in its endogenous light chain (κ) gene (as described by Chen et al., EMBO J. 12, 821-830 (1993)) and a CMD disruption in its endogenous heavy chain gene (as described in Example 1 of WO 01 / 14424). The KCo5 human κ light chain transgene is described by Fishwild et al., Nature Biotechnology 14, 845-851 (1996)), and the HCo7 human heavy chain transgene is described by US Pat. No. 5,770,429.

[0794] The Hco12 mouse has a JKD disruption in its endogenous light chain (κ) gene (as described by Chen et al., EMBO J. 12, 821-830 (1993)) and a CMD disruption in its endogenous heavy chain gene (as described in Example 1 of WO 01 / 14424). The KCo5 human κ light chain transgene is described by Fishwild et al., Nature Biotechnology 14, 845-851 (1996), and the HCo12 human heavy chain transgene is described in Example 2 of WO 01 / 14424. In the KM mouse strain, the mouse endogenous κ light chain gene has been homozygously disrupted as described by Chen et al., EMBO J. 12, 811-820 (1993), and the endogenous mouse heavy chain gene has been homozygously disrupted as described in Example 1 of WO 01 / 09187. This mouse strain carries the human kappa light chain transgene KCo5 as described in Fishwild et al., Nature Biotechnology 14, 845-851 (1996). This mouse strain also carries a human heavy chain transchromosome consisting of chromosome 14 fragment hCF (SC20) as described in WO 02 / 43478.

[0795] KM mice contain a human heavy chain transchromosome and a human kappa light chain transgene. In KM mice, the endogenous mouse heavy and light chain genes are also disrupted, so that immunization of the mice produces human, rather than mouse, immunoglobulins. The construction of KM mice and their use in producing human immunoglobulins are described in detail in WO 02 / 43478.

[0796] Splenocytes from these transgenic mice can be used to produce hybridomas that secrete human monoclonal antibodies according to well-known techniques. These transgenic mammals, mammals containing nucleic acid sequences encoding operable for expressing CD38BP, mammals stably transfected with one or more nucleic acid sequences encoding CD38, etc. are further features of the present invention.

[0797] Human monoclonal or polyclonal antibodies of the present invention, or antibodies derived from other species of the present invention, can also be produced by transgenic means in another non-human animal or plant offspring, wherein the non-human mammal or plant is genetically modified with the immunoglobulin heavy and light chain sequences of interest and can produce the antibody in a recyclable form. In connection with transgenic production in mammals, antibodies can be produced in a recyclable form in the milk of sheep, cattle, or other mammals. See, for example, US 5,827,690, US 5,756,687, US 5,750,172, and US 5,741,957.

[0798] In addition, human antibodies of the present invention or antibodies of the present invention derived from other species can be produced by display-based technologies using techniques well known in the art, including but not limited to phage display, retroviral display, ribosome display and other technologies, and the resulting molecules can be used for additional maturation such as affinity maturation. Such technologies are well known in the art (see, for example, Hoogenboom et al., J. Mol. Biol. 227, 381 (1991) (phage display), Vaughan et al., Nature Biotech 14, 309 (1996) (phage display), Hanes and Plucthau, PNAS USA 94, 4937-4942 (1997) (ribosome display), Parmley and Smith, Gene 73, 305-318 (1988) (phage display), Scott TIBS 17, 241-245 (1992), Cwirla et al., PNAS USA 87, 6378-6382 (1990), Russet et al., Nucl. Acids Research 21, 1081-1085 (1993), Hoogenboom et al., Immunol. Reviews 130, 43-68 (1992), Chiswell and McCafferty TIBTECH 10, 80-84 (1992), and US 5,733,743). If display technology is used to generate non-human antibodies, such antibodies can be humanized, for example, as described elsewhere herein.

[0799] Humanized monoclonal antibodies of the present invention can be produced by fusing the constant domains of human antibodies with the variable domains of non-human species. Examples of how to prepare humanized monoclonal antibodies can be found in, for example, US 6,054,297, US 5,886,152 and US 5,877,293. Humanized antibodies are designed to have higher homology with human immunoglobulin than monoclonal antibodies of animal origin. Non-human amino acid residues from "input" (animal) variable domains are typically transfected into people's "skeleton". In essence, humanization can be performed by replacing the complement determining regions ("CDRs") or CDR sequences of mice with the corresponding sequences of human antibodies according to the method of Winter and colleagues (Jones et al., Nature 321, 522-525 (1986), Riechmann et al., Nature 332, 323-327 (1988), Verhoeyen et al., Science 239, 1534-1536 (1988)). Thus, in such "humanized" antibodies, the CDR portion of the human variable domain has been replaced by the corresponding portion of a non-human species. Thus, humanized antibodies are typical human antibodies in which some CDR residues and possibly some framework residues are replaced by analogous sites in rodent antibodies. The choice of human heavy and light chain variable domains used to prepare humanized antibodies is crucial to reduce antigenicity. According to the so-called "best fit" method, the sequence of the variable domain of a rodent antibody is screened against a complete library of known human variable domain sequences. Then, the human sequence closest to the rodent sequence is used as the human framework (FR) of the humanized antibody (Sims et al., J. Immunol., 151, 2296 (1993), Chothia et al., J. Mol. Biol. 196, 901 (1987)). Another approach uses a specific framework derived from the consensus sequence of all human antibodies in a specific light chain or heavy chain subgroup. The same framework can be used for several different humanized antibodies (Carter et al., PNAS USA 89, 4285 (1992), Presta et al., J. Immunol. 151, 2623 (1993)).

[0800] Typically, it is also important that humanized antibodies maintain high affinity for antigens and other favorable biological properties. To achieve this goal, humanized antibodies can be prepared using a three-dimensional model of the parent sequence and the humanized sequence by analyzing the parent sequence and a variety of humanized products related thereto. Usually, a three-dimensional immunoglobulin model is available, and those skilled in the art are more aware of this. There are computer programs that can illustrate and display the possible three-dimensional conformational structures of selected candidate immunoglobulin sequences. Investigating these displayed structures can analyze the possible effects of specific residues in the function of the candidate immunoglobulin sequence, that is, analyzing the ability that can influence the candidate immunoglobulin and its antigen binding. Although CDR residues directly and substantially affect antigen binding, in this way, FR residues can be selected and combined from the receptor, and the sequence can be imported so that the desired antibody characteristics such as increasing affinity for the target antigen can be maximized.

[0801] Any suitable technique can be used to humanize or primatize murine antibodies or antibodies from other species, and a variety of suitable techniques are well known in the art (see, for example, Winter and Harris Immunol Today 14, 43-46 (1993) and Wright et al., Crit. Reviews in Immunol. 125-168 (1992)). The desired antibody can be engineered by recombinant DNA technology to replace the C sequence with the corresponding human sequence. H 1. C H 2. C H 3. Hinge domain and / or framework domain (see WO 92 / 02190 and US 5,530,101, US 5,585,089, US 5,693,761, US 5,693,792, US 5,714,350 and US 5,777,085).

[0802] Antibodies can also be humanized by replacing rodent CDRs and CDR sequences with corresponding human antibody sequences according to the method of Winter and colleagues (Jones et al., Nature 321, 522-525 (1986), Riechmann et al., Nature 332, 323-327 (1988), Verhoeyen et al., Science 239, 1534-1536 (1988)). Thus, such "humanized" antibodies are, in a sense, chimeric antibodies (US 4,816,567), in which substantially less than an intact human variable region domain has been replaced by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues, and possibly some FR residues, are substituted by residues from homologous sites in rodent antibodies.

[0803] In addition, the use of Ig cDNA for the construction of chimeric immunoglobulin genes is known in the art (see, for example, Liu et al., PNAS USA 84, 3439 (1987) and J. Immunol. 139, 3521 (1987)). mRNA is isolated from hybridomas or other antibody-producing cells and used to produce cDNA: the target cDNA can be amplified by polymerase chain reaction using specific primers (US 4,683,195 and US 4,683,202). Optionally, a library is prepared and screened to isolate the target sequence. The DNA sequence encoding the variable region is then fused to the human constant region sequence. Human constant region (and variable region) sequences can be found in Kabat et al., (1991), and more recent relevant data can be found in http: / / www.biochem.ucl.ac.uk / ~ martin / abs / Generallnfo.html Typically, the selection of an isotype is guided by the desired effector function, such as complement fixation or antibody-dependent cytotoxicity activity. Exemplary isotypes include IgG1, IgG2, IgG3, and IgG4. Either the human light chain constant region, kappa or lambda, can be used. The chimeric humanized antibody can then be expressed by conventional methods.

[0804] In terms of multimers, the CD38BPs of the present invention may be in any suitable form. Anti-CD38 antibodies and antibody fragments, if not in a higher multimeric form, such as antibodies associated with IgM antibodies, are at least heterotrimeric. In other embodiments, CD38BP may exist in dimer or monomeric form. For example, monomeric CD38BPs of the present invention may be modified by any suitable technique to form multimeric peptide components.

[0805] If desired, the class of the anti-CD38 antibodies of the present invention can be switched by known methods. For example, an antibody of the present invention that is initially IgM can be converted to an IgG antibody of the present invention by class switching. In addition, class switching techniques can be used to convert one IgG subclass to another, for example, from IgG1 to IgG2. Thus, the effector function of an antibody of the present invention can be altered by isotype switching to, for example, IgG1, IgG2, IgG3, IgG4, IgD, IgA, IgE, or IgM antibodies for various therapeutic applications.

[0806] In one embodiment, the antibody of the present invention is an IgG1 antibody, such as an IgG1, kappa or IgG1, lambda isotype. In another embodiment, the antibody of the present invention is an IgG3 antibody, such as an IgG3, kappa or IgG3, lambda isotype. In another embodiment, the antibody of the present invention is an IgG4 antibody, such as an IgG4, kappa or IgG4, lambda isotype. In another embodiment, the antibody of the present invention is an IgA1 or IgA2 antibody. In another embodiment, the antibody of the present invention is an IgM antibody.

[0807] Anti-CD38 antibodies can be obtained from recombinant combinatorial antibody libraries such as scFv phage display libraries, which are human V-terminal fragments prepared using mRNA derived from human lymphocytes. L and V HcDNAs. Methods for preparing and screening such libraries are known in the art. A variety of commercial kits are available for generating phage display libraries. Still other methods and reagents for generating and screening antibody display libraries are available (see, e.g., U.S. Pat. No. 5,223,409, WO 92 / 18619, WO 91 / 17271, WO 92 / 20791, WO 92 / 15679, WO 93 / 01288, WO 92 / 01047, WO 92 / 09690, Fuchs et al., Bio / Technology 9, 1370-1372 (1991), Hay et al., Hum. Antibod. Hybridomas 3, 81-85 (1992), Huse et al., Science 246, 1275-1281 (1989), McCafferty et al., Nature 348, 552-554 (1990), Griffiths et al., EMBO J 12, 725-734 (1993), Hawkins et al., J. MoI. Biol. 226, 889-896 (1992), Clackson et al., Nature 352, 624-628 (1991), Gram et al., PNAS USA 89, 3576-3580 (1992), Garrad et al., Bio / Technology 9, 1373-1377 (1991), Hoogenboom et al., Nuc Acid Res 19, 4133-4137 (1991) and Barbas et al., PNAS USA 88, 7978-7982 (1991). Any suitable method can be used to screen for suitable V L and V H For example, V can be screened by the antigenic determinant blotting method described in WO 93 / 06213. L and V HNucleic acid. Antibody libraries such as scFv libraries (using peptides containing human CD38 as antigens) can be prepared and screened using known suitable methods such as those described in, for example, WO92 / 01047, McCafferty et al., Nature 348, 552-554 (1990) and Griffiths et al., EMBO J 12, 725-734 (1993). Such antibody libraries and other combinations of CD38BPs (libraries, gene libraries, etc.) are a feature of the present invention and can be used for medical purposes to provide a more comprehensive immune response; in screening methods for immunogenic peptides, small molecules, other anti-CD38 antibodies (e.g., by competition assays); and / or as tools in diagnostic methods and components (e.g., immunoassay chips containing arrays of such antibodies that can be linked to other antibodies can be prepared by standard techniques). When the initial human V L and V H After fragmentation, a "mix and match" experiment can be performed in which the initially selected V fragments are screened for binding to the CD38-containing peptide. L and V H Different pairs of fragments to select the desired V L / V H Paired combinations. For example, the reactivity of the peptide can be determined by ELISA or other suitable antigenic determinant analysis methods (see, for example, Scott, JK and Smith, GPScience 249, 386-390 (1990), Cwirla et al., PNAS USA 87, 6378-6382 (1990), Felici et al., J. MoI. Biol. 222, 301-310 (1991) and Kuwabara et al., Nature Biotechnology 15, 74-78 (1997), which discuss these techniques and principles). Antibodies can be selected by their affinity for the antigen and / or their kinetics (dissociation rate) of dissociation from the antigen (see, for example, Hawkins et al., J. MoI. Biol. 226, 889-896 (1992)).

[0808] To further improve the quality and / or diversity of anti-CD38 antibodies, V L / V H Right V L and V H The fragment can be found in, for example, V H and / or V L Random mutations were performed in the CDR3 region of the antibody in a process similar to the in vivo somatic mutation process responsible for antibody affinity maturation in the natural immune response.H CDR3 or V L CDR3 complementary PCR primers amplify V H and V L This is achieved by using a primer that typically "introduces" a random mixture of four nucleotide bases at a specific site, so that the resulting PCR product is encoded in the V H and / or V L Random mutations were introduced into the CDR3 region. H and V L The randomly mutated V fragments can be screened again for binding to CD38-containing peptides. H and V L fragment.

[0809] After screening, nucleic acid encoding the selected antibody can be recovered from the display package (e.g., from the phage genome) and subcloned into a suitable vector using standard recombinant DNA techniques. If desired, the antibody-encoding nucleic acid can be further manipulated to generate other antibody forms and CD38BPs. To express the recombinant antibodies isolated by screening the combinatorial library, the nucleic acid containing the antibody-encoding sequence is typically cloned into a recombinant expression vector and transferred into a suitable host cell (mammalian cells, yeast cells, etc.) under conditions suitable for expression of the nucleic acid and production of the antibody.

[0810] High affinity antibody peptides such as human single-chain Fv (scFv) and Fab antibody fragments can also be isolated from such libraries using panning techniques in which the target antigen is immobilized on a solid surface such as a microtiter plate or beads (see, for example, Barbas and Burton, Trends. Biotechnol. 14, 230-234 (1996) and Aujame et al., Hum. Antibodies 8, 155-68 (1997)). Phage display of large natural libraries also makes it possible to directly isolate human antibodies without immunization (see, for example, de Haard et al., J. Biol. Chem. 274 (26), 18218-18230 (1999)).

[0811] In one embodiment, the present invention provides variant anti-CD38 antibodies. "Variant" anti-CD38 antibodies are antibodies that differ from the parent antibody (typically generated by immune reaction) in CDRs or other V H and / or V L Antibodies with one or more amino acid changes, substitutions, deletions, insertions, or additions to terminal sequences (even if such changes do not improve the antigenic determinant binding properties of the parent antibody, as long as a minimum substantial amount of the antigenic determinant binding properties of the parent antibody is retained).

[0812] Variant forms of antibody variants can be generated in each of the framework regions, constant domains and / or variable regions (or any one or more CDRs thereof) in a single variant antibody. Alternatively, mutations can be generated in only one of the antibody framework regions, variable regions (or a single CDR thereof), or constant domains. Alanine scanning mutagenesis techniques such as those described by Cunningham and Wells, Science 244, 1081-1085 (1989) can be used to generate variant V-containing antibodies. L 、V H Suitable residues for substitution or deletion can be identified in CD38BPs containing or replacing CDR sequences, although other suitable mutagenesis techniques can also be used. Single amino acid substitutions can also be generated and tested using known mutagenesis and screening methods such as those described in Reidhaar-Olson and Sauer, Science 241, 53-57 (1988) or Bowie and Sauer, PNAS USA 86, 2152-2156 (1989).

[0813] Thus, for example, in an antibody variant, one or a single amino acid residue can be introduced or inserted into, or immediately adjacent to, one or more hypervariable regions of the parent antibody, such as in one or more CDRs. Anti-CD38 antibody variants can contain any number of inserted amino acid residues, as long as the minimum substantial amount of the antigenic determinant binding characteristics of the parent antibody is retained. For example, an anti-CD38 antibody variant of the present invention can contain about 1-30 inserted amino acid residues, such as about 1-10, such as, for example, about 2-10, such as 2-5, or such as about 1-5 inserted amino acid residues. Similarly, for example, an anti-CD38 antibody variant of the present invention can contain about 1-30 deleted amino acid residues, such as about 1-10, such as, for example, about 2-10, such as 2-5, or such as about 1-5 deleted amino acid residues. Likewise, for example, anti-CD38 antibody variants of the present invention may contain about 1-30 substituted amino acid residues, such as about 1-10, such as about 2-10, such as 2-5, or such as about 1-5 substituted amino acid residues. Likewise, for example, anti-CD38 antibody variants of the present invention may contain about 1-30 additional terminal sequence amino acid residues, such as about 1-10, such as about 2-10, such as 2-5, or such as about 1-5 additional terminal sequence amino acid residues. Antibody variants of the present invention also include combinations of two or more such insertions, deletions, substitutions, and additional terminal sequence amino acid residues, as long as such variants possess at least a substantial portion of the affinity, specificity, and / or selectivity of the parent antibody for one or more CD38 antigenic determinants.

[0814] Considerations in selecting antibody variants (e.g., conservation of amino acid residue functional properties, conservation of amino acid residues based on hydropathic properties, and / or conservation of amino acid residues based on molecular weight / size) are described elsewhere herein. Typically, amino acid sequence changes such as conservative substitution variations are not expected to substantially alter the structural properties of the parent sequence (e.g., the substituted amino acids should not disrupt the secondary structure that is a functional characteristic of the parent sequence). Examples of known polypeptide secondary and tertiary structures are described in, for example, Proteins, Structures and Molecular Principles (Creighton, Ed., WH Freeman and Company, New York (1984)), Introduction to Protein Structure (C. Branden and J. Tooze, eds., Garland Publishing, New York, NY (1991)) and Thornton et al., Nature 354, 105 (1991). Further principles for the design and construction of peptide variants are discussed in, for example, Collinet et al., J Biol Chem 225(23), 17428-33 (2000).

[0815] Amino acid sequence variants of antibodies can be obtained by introducing appropriate nucleotide changes into the antibody encoding nucleic acid (e.g., by site-directed mutagenesis) or by chemical peptide synthesis. Such variants include, for example, deletions and / or insertions and / or substitutions and / or terminal sequence additions of residues in the antibody amino acid sequences exemplified herein. Any combination of deletions, insertions, and substitutions can be generated to obtain the desired variant, as long as the variant has a minimum substantial portion of the antigenic determinant binding properties of the parent antibody. Changes in the amino acid sequence relative to the parent antibody can also alter the post-translational processing of the variant antibody relative to the parent antibody, for example, by altering the number or position of glycosylation sites.

[0816] Variant antibodies of the present invention may comprise alterations in hypervariable regions such as CDRs. Examples of CD38BPs containing such CDR variants are described elsewhere herein, and as described above, such CD38BPs may be antibodies.

[0817] The variant antibodies of the present invention may comprise framework (FR) changes outside of the hypervariable region, for example, in the Fc region, which may be associated with advantageous properties such as altered antibody function or pharmacokinetic properties. For example, substitutions or other modifications (insertions, deletions, terminal sequence additions, or any combination thereof) in the framework region or constant domain are associated with increased half-life of the variant antibody relative to the parent antibody, or may be used to alter the immunogenicity of the variant antibody relative to the parent antibody, to provide a site for covalent or non-covalent binding to another molecule, or to alter properties such as complement binding, such as resulting in a decrease or increase in C1q binding and CDC or FcγR binding and antibody-dependent cytotoxicity (ADCC). For example, substitutions may be made at one or more amino acid residues at positions 234, 235, 236, 237, 297, 318, 320, and 322 of the heavy chain constant region, thereby resulting in changes in effector function while maintaining antigen binding compared to the unmodified antibody, see US 5,624,821 and US 5,648,260. Further references include WO 00 / 42072, which discloses that antibodies with altered Fc regions have increased ADCC, and WO 94 / 29351, which discloses that antibodies with altered Fc regions have increased ADCC. H Antibodies with mutations in the N-terminal region of the FcγRIII domain alter their ability to bind to FcRI, thereby reducing their ability to bind to C1q, which in turn reduces their ability to bind complement. Furthermore, Shields et al., J. Biol. Chem. 276, 6591-6604 (2001) describe combinatorial variants that improve FcγRIII binding to, for example, T256A / S298A, S298A / E333A, and S298A / E333A / K334A.

[0818] The in vitro half-life of an antibody can also be improved by altering the rescue receptor epitope of the Ig constant domain or Ig-like constant domain so that the molecule does not contain a complete C H 2 domains or a complete Ig Fc region, see US6,121,022 and US6,194,551. The in vitro half-life can be increased further by introducing mutations into the Fc region, such as substitution of threonine at position 252 for leucine, threonine at position 254 for serine, and threonine at position 256 for phenylalanine, see US6,277,375.

[0819] In one embodiment, the present invention provides variant anti-CD38 antibodies in which potential T cell antigenic determinants in the antibody are reduced or deleted by conventional design. Thus, for example, in one embodiment, the present invention provides "deimmunized" anti-CD38 antibodies in which potential T cell antigenic determinants are deleted. The design and construction of deimmunized anti-CD38 antibodies can be achieved by any suitable known technique (see, for example, WO9852976 for methods for preparing deimmunized antibodies). When such CD38BPs (e.g., anti-CD38 variant antibodies) as described herein are administered, it is expected that immunogenicity in humans will be eliminated or substantially reduced.

[0820] Other framework mutations include sequence changes that reduce susceptibility to proteolysis, reduce susceptibility to oxidation, and / or confer or alter other physiochemical or functional properties in the variant antibody. Alterations in the framework amino acid sequence can also result in altered glycosylation patterns in the variant antibody relative to the parent antibody. Alterations involve the deletion of one or more carbohydrate moieties found in the parent antibody and / or the addition of one or more glycosylation sites not present in the parent antibody. Antibody glycosylation is typically N-linked or O-linked. N-linked refers to the attachment of a carbohydrate moiety to the side chain of an aspartic acid residue. The tripeptide sequences aspartic acid-X-serine and aspartic acid-X-threonine are common recognition sequences for enzymatic attachment of a carbohydrate moiety to the aspartic acid side chain, where X is any amino acid except proline. Therefore, the presence of these two tripeptide sequences in a polypeptide represents a potential glycosylation site. O-linked glycosylation refers to the attachment of sugars such as N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, but 5-hydroxyproline or 5-hydroxylysine may also be used. Glycosylation sites can be added to the antibody by altering the amino acid sequence to contain one or more of the above-mentioned tripeptide sequences (for N-linked glycosylation sites). Alternatively, the original antibody sequence can be altered by adding or replacing one or more serine or threonine residues (for O-linked glycosylation).

[0821] Antibodies can also be expressed in transfectomas without the addition of the trehalose units typically attached to the carbohydrate linked to Asn at position 297 of the Fc to enhance the affinity of the Fc for FcγRIII, thereby resulting in enhanced ADCC of the antibody in the presence of NK cells, see Shield et al., J. Biol. Chem. 277, 26733 (2002). Other methods for altering glycosylation based on trehalose are described in WO 00 / 61739 by Kyowa. In addition, glycosylation can be altered to alter CDC. Further references include WO 99 / 54342 and Umana et al., Nat. Biotechnol. 17, 176 (1999), which describe the expression of monoclonal antibodies with altered glycoforms and improved ADCC activity, which were generated by constructing a CHO cell line expressing GntIII.

[0822] Other possible suitable techniques for preparing novel anti-CD38 antibodies include CDR walking mutagenesis, antibody chain replacement, "minimalist mutagenesis" (Balint and Larrick, Gene 137, 109-118 (1993)), and other affinity maturation techniques (see, e.g., Wu et al., PNAS USA 95, 6037-42 (1998)). Antibody library cloning techniques can also be used to generate variant antibodies (see, e.g., WO 96 / 33279).

[0823] There are a variety of techniques known for generating CDR variants, and any suitable technique or combination thereof can be used in the present invention to generate CDR variants of the antibody CDRs of the embodiments. Examples of such techniques include deletion of non-essential residues, as described in Studnicka et al., Protein Engineering 7, 805-814 (1994) (see also Soderlind et al., Immunotechnology. 4(3-4), 279-85 (1999), CDR walking mutagenesis and other artificial affinity maturation techniques (see, for example, Yang et al., Journal of Molecular Biology 254(3). 392-403 (1995)), CDR replacement techniques, in which CDRs are typically amplified from a different set of gene templates containing selectively synthesized oligonucleotides, and the V L 、V HThe constant regions and / or CDRs of the parental framework are then mixed (in single-stranded or double-stranded form) and assembled by polymerase chain reaction (PCR) to produce a set of antibody fragments encoded by gene products carrying the replaced CDRs introduced into the parental framework, the gene products being amplified with outer primers that anneal to sites outside the insertion restriction sites to ensure the production of full-length products, which are inserted into a vector selected from the group consisting of for expression of proteins containing variant CDRs. The appropriate structure can be determined by superimposing the structure of the variant / mock structure and the parental sequence, for example, by comparing NMR solution structures. Useful methods for the rational design of CDR sequence variants are described, for example, in WO 91 / 09967 and WO 93 / 16184. Other embodiments of these methods are provided elsewhere herein.

[0824] The present invention also provides fragments (CD38 binding fragments) of the antibodies of the present invention (including variant antibodies) that have the ability to bind to CD38. Therefore, CD38BPs include antibody-like molecules that contain less than the entire tetrameric structure of a natural antibody. Antibody fragments can be any peptide containing a portion of a full-length antibody, typically an antigen-binding or variable region portion thereof (this includes, for example, humanized antibody fragments containing the CDRs of the antibodies of the present invention, variants thereof, or other CDRs that allow the antigen fragment to compete with the antibodies of the present invention for binding to CD38). In one embodiment, an antibody fragment refers to a peptide consisting mainly of or only of a portion of an antibody molecule. In one embodiment, the present invention provides a peptide containing at least one or more V-terminal fragments of the present invention. H The heavy chain variable domain of the CDRs optionally also contains one or more V L In some embodiments, the antibody fragment comprises a portion of a light chain variable domain of CDRs, wherein the heavy chain variable domain and optionally the light chain variable domain are selectively fused to other portions, such as immunoglobulin constant domains. The constant domain sequence can be added to the heavy chain and / or light chain sequence to form a species with a partial length heavy chain and / or light chain. Any antibody isotype constant region or portion thereof, including the constant regions of IgG, IgM, IgA, IgD, and IgE, can be used to achieve this purpose.

[0825] Examples of CD38-binding antibody fragments include Fab, Fab', F(ab')2, and Fv fragments. Antibody fragments of the present invention also include peptides containing CDRs, etc. In one embodiment, the present invention provides a first polypeptide chain comprising any of the heavy chain CDRs described herein and a second polypeptide chain comprising any of the light chain CDRs described herein, wherein the two polypeptide chains are covalently linked by one or more intrachain disulfide bonds. In one embodiment, the present invention provides a two-chain antibody fragment having such characteristics, wherein the antibody fragment is selected from Fab, Fab', Fab'-SH, Fv, and / or F(ab')2 fragments. Antibodies can be fragmented by conventional methods, and the fragments can be screened using the same methods described above for whole antibodies. For example, F(ab')2 fragments can be produced by treating antibodies with pepsin. The resulting F(ab')2 fragments can be treated to remove disulfide bonds, thereby producing Fab1 fragments. Fab fragments can be obtained by treating IgG antibodies with papain; Fab' fragments can be obtained by digesting IgG antibodies with pepsin. Fab1 fragments can be produced by linking Fab1 via sulfhydryl groups or disulfide bonds as described below. Fab1 fragments are antibody fragments obtained by cleaving the disulfide bonds of the hinge region of F(ab')2. Fab' fragments can be obtained by treating F(ab')2 fragments with a reducing agent, such as dithiothreitol. Antibody fragment peptides can also be produced by expressing nucleic acids encoding such peptides in recombinant cells (see, for example, Evans et al., J. Immunol. Meth. 184, 123-38 (1995)). For example, a chimeric gene encoding a portion of a F(ab')2 fragment may include a C H 1 domain and the H chain hinge region, followed by a translation stop codon to produce this truncated antibody fragment molecule.

[0826] CD38BPs also include monovalent antibodies and single-chain antibodies. Single-chain antibodies are peptides in which the heavy and light chain Fv regions are linked. In one embodiment, the present invention provides single-chain Fvs (scFvs), in which the heavy and light chains in the Fv of the anti-CD38 antibody of the present invention are linked to a single peptide chain via a flexible peptide linker (typically about 10, 12, 15 or more amino acid residues). Methods for producing such antibodies are described, for example, in US 4,946,778, Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994), Bird et al., Science 242, 423-426 (1988), Huston et al., PNAS USA 85, 5879-5883 (1988), and McCafferty et al., Nature 348, 552-554 (1990). H and V L , then the single-chain antibody can be monovalent, if two V H and V L , it is bivalent. If more than two V H and V L , it is multivalent.

[0827] In one embodiment of the present invention, CD38BP can be derived from or linked to another functional molecule, such as another peptide or protein (e.g., a Fab' fragment), thereby generating a bispecific or multispecific molecule that binds to multiple binding sites or target antigenic determinants. For example, an antibody of the present invention can be functionally linked to one or more other binding molecules, such as another antibody, peptide, or binding analog (e.g., by chemical coupling, genetic fusion, non-covalent linkage, or other means). In one embodiment, CD38BP is an antibody of the present invention.

[0828] Thus, the present invention includes bispecific and multispecific molecules comprising at least one first binding specificity for CD38 and a second binding specificity for a second target antigenic determinant. In one embodiment of the present invention, the second target antigenic determinant is an Fc receptor, such as human FcγRI (CD64) or human Fcα receptor (CD89), or a T cell receptor, such as CD3. In one embodiment, the present invention provides bispecific and multispecific molecules that can bind to effector cells expressing FoγR, FcαR, or FcεR (e.g., monocytes, macrophages, or polymorphonuclear leukocytes (PMNs)) and act on cells expressing CD38. These bispecific and multispecific molecules localize CD38-expressing cells to effector cells and initiate Fc receptor-mediated effector cell activities such as phagocytosis of CD38-expressing cells, antibody-dependent cellular cytotoxicity (ADCC), cytokine release, or superoxide anion production.

[0829] In addition to the anti-Fc binding specificity and the anti-CD38 binding specificity, the bispecific and multispecific molecules of the present invention may further include a third binding specificity. In one embodiment, the third binding specificity is an anti-enhancement factor (EF) portion, for example, a molecule that binds to a surface protein and participates in cytotoxic action, thereby increasing the immune response to the target cell. The "anti-enhancement factor portion" can be an antibody, functional antibody fragment or ligand that binds to a given molecule, for example, an antigen or receptor, thereby resulting in an enhancement effect of the binding determinant on the Fc receptor or target cell antigen. The "anti-enhancement factor portion" can bind to an Fc receptor or a target cell antigen. Optionally, the anti-enhancement factor portion can be combined with an entity different from the entity that can bind to the first and second binding specificities. For example, the anti-enhancement factor portion can bind to a cytotoxic T cell (for example, through CD2, CD3, CD8, CD28, CD4, CD40, ICAM-1 or other immune cells that have a higher immune response against the target cell).

[0830] In one embodiment, the bispecific and multispecific molecules of the present invention comprise at least one other antibody having binding specificity, such as Fab, Fab1, F(ab')2, Fv, or scFv. The other antibody may also be a light chain or heavy chain dimer, or any minimal fragment such as an Fv or a single-chain construct as described by Ladner et al. in US 4,946,778. The antibody may also be a binding domain immunoglobulin fusion protein as described in US 2003 / 0118592 and US 2003 / 0133939.

[0831] In one embodiment, binding specificity for an Fc receptor is provided by a human monoclonal antibody whose binding is not blocked by human immunoglobulin G (IgG). The term "IgG receptor" as used herein refers to any of the eight gamma chain genes located on chromosome 1. These genes encode a total of 12 transmembrane or soluble receptor isoforms that can be classified into three Fc* receptor classes: FcγRI (CD64), FcγRII (CD32), and FcγRIII (CD16). In one embodiment, the Fcγ receptor is human high-affinity FcγRI. Fanger et al. describe the generation and characterization of these monoclonal antibodies in WO 88 / 00052 and US 4,954,617. These antibodies bind to antigenic determinants of FcγRI, FcγRII, or FcγRIII at a site distinct from the Fcγ binding site of the receptor, and therefore, their binding is not completely blocked by physiological levels of IgG. The specific anti-FcγRI antibodies used in the present invention include mAb 22, mAb 32, mAb 44, mAb 62, and mAb 197. In other embodiments, the anti-Fcγ receptor antibody is a humanized form of mAb 22 (H22). The production and properties of the H22 antibody are described in Graziano, RF et al., J. Immunol. 155(10), 4996-5002 (1995) and WO 94 / 10332. The cell line producing the H22 antibody was deposited with the American Type Culture Collection on November 4, 1992, under the designation HA022CL1 and has the sequence number CRL11177.

[0832] In one embodiment, Fc receptor binding specificity is provided by an antibody that binds to a human IgA receptor, such as the Fcα receptor (Fcα1 (CD89)), and in one embodiment, its binding is not blocked by human immunoglobulin A (IgA). The term "IgA receptor" refers to a gene product containing an α gene (FcαRI) located on chromosome 19. This gene is known to encode several alternatively spliced ​​transmembrane isoforms of 55 to 110 kDa. FcαRI (CD89) is constitutively expressed on monocytes / macrophages, eosinophils, and neutrophils, but not on non-effector cell populations. FcαRI has an intermediate affinity for IgA1 and IgA2, and its affinity is increased upon contact with cytokines such as G-CSF or GM-CSF (Morton, HC et al., Critical Reviews in Immunology 16, 423-440 (1996)). Four FcαRI-specific monoclonal antibodies, identified as A3, A59, A62, and A77, that bind FcαRI outside of the IgA ligand binding domain have been described (Monteiro, RC et al., J. Immunol. 148, 1764 (1992)).

[0833] FcαRI, FcγRI, FcγRII and FcγRIII, particularly FcγRII and FcγRIII, are examples of triggering receptors used in the present invention because they (1) are primarily expressed on immune effector cells, such as monocytes, PMNs, macrophages and dendritic cells; (2) are expressed at high levels (e.g., 5,000-100,000 per cell); (3) are mediators of cytotoxic effects (e.g., ADCC, phagocytosis); and (4) mediate enhanced antigen presentation to them, including self-antigens.

[0834] In one embodiment, the CD38BP of the present invention is a multispecific anti-CD38 antibody or antibody-like molecule, and a specific embodiment thereof comprises at least one pair of V domains specific for an antigenic determinant containing at least a portion of CD38. H Sequence and V L sequence chain, and at least one pair of V H and V L The V in bispecific antibodies H and V L The sequence may comprise a sequence corresponding to anti-CD38 V H and V L Region, variant V H and / or V L Sequence, or V Hand / or V L Complete V of the appropriate part of the zone H and V L Sequences, for example, CDR sequences and other sequences sufficient to provide binding to the desired antigenic determinant are appropriately combined.

[0835] Exemplary bispecific antibody molecules include (i) two antibodies linked to each other, one specific for CD38 and the second for a second target, (ii) a single antibody having one chain specific for CD38 and a second chain specific for a second molecule, and (iii) a single chain antibody specific for CD38 and a second molecule. Typically, the second target / second molecule is a molecule other than CD38. In one embodiment, the second molecule is a cancer antigen / tumor-associated antigen, such as carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), RAGE (renal antigen), α-fetoprotein, CAMEL (an antigen that recognizes CTLs on melanoma), CT antigens (such as MAGE-B5, -B6, -C2, -C3, and D; Mage-12; CT10; NY-ESO-1, SSX-2, GAGE, BAGE, MAGE, and SAGE), mucin antigens (such as MUC1, mucin CA125, etc.), ganglioside antigens, tyrosinase, gp75, C-myc, Marti, MelanA, MUM-1, MUM-2, MUM-3, HLA-B7, and Ep-CAM. In one embodiment, the second molecule is a cancer-associated integrin such as α5β3 integrin. In one embodiment, the second molecule is an angiogenic growth factor or other cancer-related growth factor, such as vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), epidermal growth factor receptor (EGFR), angiogenin and its receptor, particularly a receptor associated with cancer progression (e.g., one of the HER1-HER4 receptors). Other proteins associated with cancer progression described herein are also suitable second molecules. In one embodiment, the second molecule is a molecule expressed on the surface of multiple myeloma cells, such as CD138.

[0836] In one embodiment, the bispecific antibody of the present invention is a bifunctional antibody.

[0837] Bispecific antibodies also include cross-linked or "heterolinker" antibodies. For example, the antibody in the heterolinker can be coupled to avidin and other molecules directed against biotin. For example, it has been proposed to use such antibodies to allow immune system cells to be directed against other cells (see, for example, US 4,676,980). Any convenient cross-linking method can be used to generate heterolinker antibodies. Suitable peptide cross-linking reagents and techniques are well known in the art, and examples of such reagents and techniques are disclosed, for example, in US 4,676,980.

[0838] Therefore, although antibodies are discussed herein, it should be understood that, where appropriate, the details and features of antibodies are equally applicable to antibody fragments such as Fab fragments, Fab1 fragments, and scFv peptides, antibody-like peptides (CDR-containing peptides), bispecific and multispecific antibodies, and other CD38BPs, as long as the CD38BPs of the present invention retain at least a substantial portion of the antigen-binding properties of the corresponding intact antibody. In some cases, antibody fragments may have lower antigen-binding affinity but may provide other advantageous features that compensate for any such loss in affinity.

[0839] The CD38BPs of the present invention, particularly anti-CD38 antibodies, can be selected based on their ability or inability to perform complement fixation. There are a variety of antibody isotypes capable of complement fixation and CDC, including, but not limited to, mouse IgM, mouse IgG2a, mouse IgG2b, mouse IgG3, human IgM, human IgG1, and human IgG3. These do not include, nor are they limited to, the isotypes mouse-human IgG2 and human IgG4. Isotype assays and other methods for altering the complement fixation and CDC functional characteristics of antibodies are known in the art. The CD38BPs of the present invention also include immunoadhesins, which are molecules to which one or more CDRs of an anti-CD38 antibody are covalently or non-covalently attached. Immunoadhesins can incorporate the CDR(s) as part of a larger polypeptide chain, can covalently attach the CDR(s) to another polypeptide chain, or can be non-covalently incorporated into the CDR(s). The CDRs enable the immunoadhesin to specifically bind to CD38.

[0840] The present invention also provides CD38BP fusion proteins. The CD38BP fusion protein may comprise any suitable amino acid sequence or combination of sequences that specifically and / or selectively targets at least a portion of at least one domain contained in CD38 (e.g., anti-CD38 antibody V H domain, V LThe fusion protein comprises a secondary sequence that is a non-homologous, typically substantially non-identical, amino acid sequence (e.g., less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20% amino acid identity to the CD38-specific / selective sequence) that confers detectable biological functions and / or properties not conferred by the CD38-specific / selective sequence alone (e.g., increased in vivo half-life, fluorescence, increased activity on specific cell types, etc.). The functional sequences of such fusion proteins can be separated by a flexible linker. The secondary sequence can also be derived from a cytotoxic or apoptotic peptide. The secondary sequence can also provide diagnostic properties. Examples of such sequences include those of readily visualized enzymes such as horseradish peroxidase.

[0841] CD38BP fusion proteins can also be described by comparing their antigenic determinant tags. An antigenic determinant tag sequence is an amino acid sequence having sufficient residues to provide an antigenic determinant for antibodies to be generated, but in the case of CD38BP, this sequence is so short that it does not substantially interfere with the activity (selectivity, specificity, affinity, and / or biological activity) of CD38BP (compared to the parent CD38BP lacking the antigenic determinant tag). The desired antigenic determinant tag should be sufficiently specific so that anti-antigenic determinant tag antibodies do not substantially cross-react with other antigenic determinants. Suitable tag polypeptides generally have at least about 6 amino acid residues, typically 8-50 amino acid residues (e.g., about 9-30 residues). Examples of antigenic determinant tags include the flu HA tag polypeptide and its antibody 12CA5 (Field et al., MoI. Cell. Biol. 8, 2159-2165 (1988)); the myc tag and its antibodies 8F9, 3C7, 6E10, G4, B7 and 9E10 (Evan et al., MoI. Cell. Biol. 5 (12), 3610-3616 (1985)), and the herpes simplex virus glycoprotein D (gD) tag and its antibodies (Paborsky et al., Protein Engineering 3 (6), 547-553 (1990)). In a specific embodiment, the antigenic determinant tag is a "rescue receptor binding antigenic determinant". The term "rescue receptor binding antigenic determinant" as used herein refers to an antigenic determinant in the Fc region of an IgG molecule (IgG1, IgG2, IgG3, or gG4), which is responsible for increasing the serum half-life of the IgG molecule in vivo.

[0842] The CD38BPs of the present invention also include CD38BP derivatives. Derivatives are peptides in which one or more amino acid residues of the peptide are chemically modified (e.g., alkylated, acylated, esterified, or amino-formed) or covalently linked to one or more heterologous surrogates (e.g., lipophilic surrogates, PEG groups, peptide side chains linked to suitable organic group linkers, etc.). Peptides can also be linked to therapeutic groups such as cytotoxins, chemotherapeutic drugs, immunosuppressants, or radioisotopes (so-called immunoconjugates). Generally, the CD38BPs described herein can be modified with substances containing any suitable number of such modified amino acids and / or linked to such linker surrogates. In this context, suitability is generally determined by the ability to at least substantially retain the CD38 selectivity and / or specificity possessed by the non-derivatized parent CD38BP. The advantages of containing one or more modified amino acids are, for example, (a) increasing the serum half-life of the polypeptide, (b) reducing the antigenicity of the polypeptide, or (c) increasing the storage stability of the polypeptide. For example, amino acids can be modified during recombinant production, during translation, or post-translationally (e.g., N-linked glycosylation on NXS / T elements when expressed in mammalian cells), or by synthetic means. Non-limiting examples of modified amino acids include glycosylated amino acids, sulfated amino acids, prenlyated (e.g., farnesylated, geranylgeranylated) amino acids, acetylated amino acids, acylated amino acids, PEGylated amino acids, biotinylated amino acids, carboxylated amino acids, phosphorylated amino acids, and the like. References guiding the skilled artisan in amino acid modification are found throughout the literature. Experimental procedures can be found in Walker (1998) Protein Protocols On CD-ROM, Humana Press, Towata, NJ. Modified amino acids can be selected from glycosylated amino acids, PEGylated amino acids, farnesylated amino acids, acetylated amino acids, biotinylated amino acids, amino acids linked to a lipid group, and amino acids linked to an organic derivatizing agent.

[0843] In addition, antibodies can be chemically modified by being covalently linked to polymers, for example, to increase their circulation half-life. Illustrated polymers and methods for linking them to peptides are described in, for example, US 4,766,106, US 4,179,337, US 4,495,285, and US 4,609,546. Other illustrative polymers include polyoxyethylated polyols and polyethylene glycol (PEG) (e.g., having a molecular weight between about 1,000 and about 40,000, such as between about 2000 and about 20,000, for example, about 3,000-12,000 PEG).

[0844] In one embodiment, the present invention provides a CD38BP linked to a second molecule selected from a radionuclide, an enzyme, an enzyme substrate, a cofactor, a fluorescent marker, a chemiluminescent marker, a peptide tag, or a magnetic particle. In one embodiment, the CD38BP can be linked to one or more antibody fragments, nucleic acids (oligonucleotides), nucleases, hormones, immunomodulators, chelating agents, boron compounds, photosensitizers, dyes, and the like. These and other suitable agents can be coupled directly or indirectly to the CD38BPs of the present invention. An example of indirect coupling to a second agent is coupling via a spacer group. These spacers can be insoluble or soluble (see, for example, Diener et al., Science 231.148 (1986)), and the spacer can be selected so that the drug is released from the CD38BP at the target site and / or under specific conditions. Other examples of therapeutic agents that can be coupled to CD38BPs include lectins and fluorescent peptides.

[0845] In one embodiment, a CD38BP derivative containing one or more radiolabeled amino acids is provided. Radiolabeled CD38BP can be used for diagnostic and therapeutic purposes (coupling with a radioactive molecule is another possible feature). Non-limiting examples of labeled polypeptides include, but are not limited to, 3 H. 14 C. 15 N. 35 S. 90 Y. 99 Tc, 125 I. 131 I and 186 Re. Methods for preparing radiolabeled amino acids and related peptide derivatives are known in the art (see, for example, Junghans et al., in Cancer Chemotherapy and Biotherapy 655-686 (2d edition, Chafner and Longo, eds., Lippincott Raven (1996)) and US 4,681,581, US 4,735,210, US 5,101,827, US 5,102,990 (US RE 35,500), US 5,648,471 and US 5,697,902. For example, radioisotopes can be coupled by the chloramine T method.

[0846] Radionuclides useful in diagnosis are indium radioisotopes, while cytotoxic yttrium radioisotopes are used therapeutically. Radioisotopes that emit proton photons are generally useful in diagnostic (radioimmunoimaging (RIS)) methods. Auger electrons have a very short path length (5-10 nm) and must be internalized to produce cytotoxicity (see, for example, Adelstein et al., Nucl. Med. Biol. 14, 165-169 (1987)). Therefore, peptides conjugated to such radioisotopes can be used in diagnostic methods, but only internalized peptides are believed to release therapeutic Auger electrons. Alpha particles need to be in close proximity to cells (within 3-4 cell diameters) to be effective therapeutic agents (Vriesendorp et al., "Radioimmunoglobulin therapy," in High Dose Cancer Therapy, Armitage et al., (eds.) (Williams & Wilkins, Baltimore, Md. 1992)). Both Auger electrons and alpha reflectors can be considered highly selective due to their short range emission, typically sparing adjacent normal cells.

[0847] radio metals 111 In and 90 Y is a pure gamma-emitter and a pure beta-emitter, respectively. The most commonly used Auger electron emitter, iodine-125, has a half-life of approximately 60 days and is typically released in vivo by immunoconjugates (due to dehalogenation). The most commonly used alpha emitters, astatine-211 and bismuth-212, have relatively short half-lives (7.2 h and 1.0 h, respectively) and decay after the first alpha emission to a radioisotope that is not retained by the immunoconjugate (Wilbur, Antibiot. Immunoconjug. Radiopharm. 4, 5-97 (1991)). For diagnostic applications, CD38BPs labeled with indium-111 or technetium-99m can be used. Both of these radioisotopes emit gamma rays within a suitable energy range (100-250 keV) for imaging. Typically, energies below this range are insufficient to penetrate and reach external imaging equipment. Higher energy levels are difficult to calibrate and ...

Claims

1. An antibody that binds to human CD38 and is encoded by (i) to (iv) below (i) human light chain and human heavy chain nucleic acids containing the nucleotide sequences shown in SEQ ID No: 1 and SEQ ID No: 6, respectively, in their variable regions; (ii) human light chain and human heavy chain nucleic acids containing the nucleotide sequences shown in SEQ ID No: 11 and SEQ ID No: 16, respectively, in their variable regions; (iii) a human light chain and a human heavy chain nucleic acid comprising in its variable region the nucleotide sequences shown in SEQ ID No: 21 and SEQ ID No: 26, respectively; or (iv) Human light chain and human heavy chain nucleic acids containing in their variable regions a nucleotide sequence obtained by subjecting the sequence shown in (i), (ii) or (iii) above to conservative sequence modifications.

2. An antibody that binds to human CD38, comprising a V having an amino acid sequence as shown in SEQ ID No: 2 L district.

3. An antibody that binds to human CD38, comprising a V having an amino acid sequence as shown in SEQ ID No: 7 H district.

4. An antibody that binds to human CD38, comprising a V having a sequence as shown in SEQ ID No: 30 H CDR3.

5. An antibody that binds to human CD38, comprising a V having an amino acid sequence as shown in SEQ ID No: 22 L district.

6. An antibody that binds to human CD38, comprising a V having an amino acid sequence as shown in SEQ ID No: 27 H district.

7. An isolated human monoclonal antibody comprising (i) Derived from human Hv1263 / 3M28 (V H I) a heavy chain variable region amino acid sequence that is a germline sequence and a light chain variable region amino acid sequence that is derived from a human L15 (VκI) germline sequence, wherein the human antibody binds to human CD38; or (ii) Derived from human V H 3-DP-47 / V3-23(V H III) a heavy chain variable region amino acid sequence of a germline sequence and a light chain variable region amino acid sequence derived from a human L6 (VκI) germline sequence, wherein the human antibody binds to human CD38.

8. A pharmaceutical composition comprising the peptide as claimed in any one of claims 1 to 7 and a pharmaceutically acceptable carrier.

9. A method for treating a disease or condition in a patient in which cells expressing CD38 are involved, the method comprising administering a peptide as claimed in any one of claims 1 to 7 to a patient in need of treatment.

10. A kit for detecting the presence of CD38 antigen or cells expressing CD38 in a sample containing the peptide as claimed in any one of claims 1 to 7.

Images