Immune modulation and treatment of solid tumors with antibodies that specifically bind CD38

Abstract

To provide methods of immunomodulation and treating patients having solid tumors with antibodies that specifically bind CD38.SOLUTION: The present invention provides a method of treating a patient having a solid tumor, including administering to a patient in need thereof a therapeutically effective amount of an antibody that specifically binds CD38 for a time sufficient to treat the solid tumor.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a method for immunomodulation and treatment of solid tumors using an antibody that specifically binds CD38. Regarding. [Background technology]

[0002] The immune system is tightly regulated by a network of costimulatory and co-inhibitory ligands and receptors. These molecules provide secondary signals for T cell activation, both positive and negative. This leads to a balanced network of signaling that enhances immunity against infection and tumors. Maximizing the immune response while limiting immunity to self (Wang et al., (Epub Mar.7,2011)J Exp Med 208(3):577~92 , Lepenies et al., (2008) Endocr Metab Immu ne Disord Drug Targets 8:279~288).

[0003] Immune checkpoint therapy for treating solid tumors promotes antitumor immune responses Y, an anti-CTLA-4 and anti-PD-1 antibody that targets co-inhibitory pathways in T cells ERVOY® (ipilimumab), KEYTRUDA® (pembro The approval of OPDIVO® (nivolumab) and OPDIVO® (pembrolizumab) has improved the clinical outcomes of patients with cancer. Anti-PD-1 / PD-L1 antibodies have been used to treat patients with multiple solid tumors. Although favorable clinical responses have been observed in patients with schizophrenia, the response rate remains quite low. Approximately 15%–20% in previously treated patients (Swaika et al., (2015) Mol Immunol doi:10.1016 / j.molimm.2 015.02.009).

[0004] Natural killer cells (NK), dendritic cells (DC) and effector T cells are potent anti- Although tumor cells can induce tumor responses, they often possess an immunosuppressive microenvironment. Myeloid-derived suppressor cells (MDSCs), regulatory T cells (Tregs), or regulatory B cells favors the development of immunosuppressive populations of immune cells, such as Bregs, thus promoting tumor immune tolerance and contribute to the failure of immunotherapy regimens in cancer patients and experimental tumor models. Summary of the Invention [Problem to be solved by the invention]

[0005] Thus, they can be used to induce adaptive immune responses against tumors or target immunosuppressive immune cells. There remains a need to develop new cancer immunotherapies that [Means for solving the problem]

[0006] The present invention provides a therapeutically effective amount of an antibody that specifically binds CD38, and The present invention provides a method of treating a patient having a solid tumor, comprising administering to the patient:

[0007] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 to a subject in need thereof. and administering to a patient suffering from a regulatory T cell (Treg) mediated disorder. Methods of treating the condition are also provided.

[0008] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 to a subject in need thereof.

[0023] The present invention relates to a method for treating a myeloid-derived suppressor cell (MDSC)-mediated disease, comprising administering the method to a patient suffering from the disease. Also provided are methods of treating patients with

[0009] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 to a subject in need thereof. and administering to a patient suffering from a regulatory B cell (Breg) mediated disease. Methods of treating the condition are also provided.

[0010] The present invention also relates to a method for the treatment of neurodegenerative diseases by contacting regulatory T cells (Treg) with an antibody that specifically binds CD38. Also provided is a method for suppressing Treg activity, comprising:

[0011] The present invention also relates to a method for treating myeloid-derived suppressor cells (MDSCs) that specifically bind CD38. Also provided is a method of inhibiting the activity of MDSCs comprising contacting with the antibody.

[0012] The present invention also relates to a method for the treatment of inflammatory bowel disease by contacting regulatory B cells (Bregs) with an antibody that specifically binds CD38. Also provided is a method for inhibiting Breg activity, comprising:

[0013] The present invention also provides a method for treating a patient comprising administering to the patient an antibody that specifically binds CD38. Also provided are methods for enhancing the immune response in a subject.

[0014] The present invention also provides a method for the treatment of a patient's inflammatory bowel disease by administering to the patient an antibody that specifically binds CD38. US20100232633A1 - Method for treating a patient with a solid tumor, comprising reducing the number of Treg cells in the patient - Google Patents Also provided.

[0015] The present invention also provides a method for the treatment of a patient's inflammatory bowel disease by administering to the patient an antibody that specifically binds CD38. and reducing myeloid-derived suppressor cell (MDSC) numbers in patients with solid tumors. Also provided are methods of treating a patient having

[0016] The present invention also relates to a method for producing an immune suppressing cell that specifically binds CD38. Also provided is a method for inhibiting the activity of immune suppressor cells, comprising contacting the immune suppressor cells with an antibody that inhibits the activity of immune suppressor cells. do.

[0017] The present invention also relates to a method for administering an antibody that specifically binds CD38 to a patient in need thereof. Also provided is a method of treating a patient having a viral infection, comprising: [Brief description of the drawings]

[0018] [Figure 1] Median lymphocyte counts increased over time in patients treated with DARZALEX™ (daratumumab) at a dose of 8 mg / kg (upper line) or 16 mg / kg (lower line), showing lymphocyte counts returning to baseline after treatment ended. Study name: SIRIUS. X-axis shows treatment cycle and time as dosing day within each treatment cycle (C1D1: cycle 1, day 1; C1D4: cycle 1, day 4, etc.). SCR: baseline, EOT: end of treatment, WK: week, POST-WK: indicated week after treatment, post-PD FU: follow-up after progression. Grey shaded highlighted areas show 25-27% interquartile range (IQR) for data points at each visit for responders. [Diagram 2] Percent change (%) from baseline in absolute CD3+ T cell counts in peripheral blood in patients treated with DARZALEX™ (daratumumab) for individual patients (light grey line). Study name: SIRIUS (MMY2002). X-axis indicates treatment cycle and time as days of administration within each treatment cycle (C1D1: cycle 1, day 1; C1D4: cycle 1, day 4, etc.). WK: weeks; POST-WK: indicated weeks after treatment; POST-PD FU: follow-up after progression. Black line indicates median % change across all patients. [Diagram 3]Percent change (%) from baseline in absolute CD4+ T cell counts in peripheral blood in patients treated with DARZALEX™ (daratumumab) for individual patients (light grey line). Study name: SIRIUS. X-axis shows treatment cycle and time as days of dosing within each treatment cycle (C1D1: cycle 1, day 1; C1D4: cycle 1, day 4, etc.). WK: weeks; POST-TMT: post-treatment. Black line shows median % change across all patients. [Figure 4] Percent change (%) from baseline in absolute CD8+ T cell counts in peripheral blood in patients treated with DARZALEX™ (daratumumab) for individual patients (light grey line). Study name: SIRIUS. X-axis shows treatment cycle and time as days of administration within each treatment cycle (C1D1: cycle 1, day 1; C1D4: cycle 1, day 4, etc.). WK: weeks; Pre-PD FU: follow-up before progression; Post-PD FU: follow-up after progression. Black line shows median % change across all patients. [Diagram 5] Figure 1 shows that CD45+CD3+ cell counts (measured as percentage of lymphocytes) in bone marrow aspirates increased over time during DARZALEX™ (daratumumab) treatment at doses of 8 mg / kg or 16 mg / kg. As indicated, the graph includes both responders and non-responders. Study name: SIRIUS. X-axis shows treatment cycle and time as dosing day within each treatment cycle (C2D22: cycle 2, day 22, etc.). SCR: baseline, Post-PD FU1: follow-up after progression. Areas highlighted in grey shades show the 25-27% interquartile range (IQR) for data points at each visit for 8 mg / kg dose non-responders, 16 mg / kg dose responders, or 16 mg / kg dose non-responders, respectively. NR: non-responders, R: responders. [Figure 6]Figure 1 shows that CD45+CD3+CD8+ cell counts (measured as percentage of lymphocytes) in bone marrow aspirates increased over time during DARZALEX™ (daratumumab) treatment at doses of 8 mg / kg or 16 mg / kg. As indicated, the graph includes both responders and non-responders. Study name: SIRIUS. X-axis shows time in terms of treatment cycle and dosing day within each treatment cycle (C2D22: cycle 2, day 22, etc.). SCR: baseline, Post-PD FU1: follow-up after progression. Areas highlighted in grey shades show the 25-27% interquartile range (IQR) for data points at each visit for 8 mg / kg dose non-responders, 16 mg / kg dose responders, or 16 mg / kg dose non-responders, respectively. NR: non-responders, R: responders. [Figure 7A] Figure 1 shows that CD8+ / Treg and CD8+ / CD4+ cell ratios in peripheral blood, expressed as median across all treated patients, increased over time during DARZALEX™ (daratumumab) treatment. Time points: C1D1: cycle 1 day; C3D1: cycle 3 day 1; C4D1: cycle 4 day 1. Study name: SIRIUS. SCR: baseline. [Figure 7B] 14. CD8+ / Treg cell ratios in bone marrow aspirates, expressed as median across all treated patients, increased over time during DARZALEX™ (daratumumab) treatment. Time points: C1D1: Cycle 1, day; C3D1: Cycle 3, day 1; C4D1: Cycle 4, day 1. Study name: SIRIUS. [Figure 8A] Demonstrating increased CD8+ T cell clonality in responders compared to non-responders as measured using % change in specific clonal cell abundance (CIA). Study name: GEN501, subset of 17 patients. [Figure 8B]Fold change in CD8+ T cell clonality of individual patients pre- and post-DARZALEX™ (daratumumab) treatment. Responders are indicated with an asterisk. Clonality was measured by fold change in abundance (CIA) of specific clonal cells. Study name: GEN501, subset of 17 patients. [Figure 8C] Compared to non-responders (Group B), responders (Group A) showed greater total expansion of the TCR repertoire as measured by CIA (change in abundance). P=0.037. Study name: GEN501, subset of 17 patients. [Figure 8D] For each expanded T cell clone, the sum of absolute change in abundance (CIA) in responders and non-responders is shown. P=0.035 between responders (group A) and non-responders (group B). Study name: GEN501, subset of 17 patients. [Figure 8E] Maximal CIA of a single T cell clone in responders (group A) and non-responders (group B). Study name: GEN501, subset of 17 patients. [Figure 8F] Compared to non-responders (group B), responders (group A) had greater maximum single clone expansion as measured by maximum CIA%. P=0.0477. Study name: GEN501, subset of 17 patients. [Figure 9A] Percentage of CD8+ naive cells in peripheral blood of non-responders (NR, black squares) and at least minimally responding patients (MR, white squares) to DARZALEX™ (daratumumab) at baseline, or at weeks 2, 4, or 8 of treatment, or after relapse. Study name: GEN501, subset of 17 patients. **p=1.82×10-4. [Figure 9B]Percentage of CD8+ central memory cells (Tem) in peripheral blood of non-responders (NR, black squares) and at least minimally responding patients (MR, white squares) to DARZALEX™ (daratumumab) at baseline, or at weeks 2, 4, or 8 of treatment, or after relapse. Study name: GEN501, subset of 17 patients. *p=4.88×10-2. [Figure 9C] Percentage increase in HLA class I restricted CD8+ T cells in peripheral blood at baseline, or at weeks 1, 4 or 8 of treatment, or after relapse. Study name: GEN501, subset of 17 patients. [Figure 9D] Low levels of CD38 expression are shown on peripheral blood CD8+ naive T cells and CD8+ central memory (Tem) cells at baseline or on treatment. Study name: GEN501, subset of 17 patients. MFI: Mean Fluorescence Intensity. [Figure 10A] Histograms of FACS analysis showing the frequency of Tregs (CD3+CD3+CD4+CD25+CD127dim (upper histogram, P4 cell population) and the frequency of CD38+ Tregs within the Treg population (lower histogram, P5 cell population) in baseline multiple myeloma patients. Study name: GEN501, subset of 17 patients. [Figure 10B] Histograms of FACS analysis showing the frequency of Tregs (CD3+CD3+CD4+CD25+CD127dim (top histogram, P4 cell population) and the frequency of CD38+ Tregs within the Treg population (bottom histogram, P5 cell population) in multiple myeloma patients following DARZALEX™ (daratumumab) treatment. DARZALEX™ (daratumumab) treatment depleted CD38+ Tregs. Study name: GEN501, subset of 17 patients. [Figure 10C]indicates frequency of CD38highCD3+CD4+CD25+CD127dim Tregs in patients treated with DARZALEX™ (daratumumab) at baseline or at end of treatment (EOT) at weeks 1, 4, 8, post-relapse, or 6 months. Frequency of CD38high Tregs decreased with DARZALEX™ (daratumumab) treatment and returned to baseline at EOT. Y-axis: % of CD38highCD3+CD4+CD25+CD127dim Tregs (derived from CD3+ T cells). Study name: GEN501, subset of 17 patients. [Figure 10D] CD8+ / Treg cell ratios in responders and non-responders at baseline, 1st, 4th and 8th weeks of treatment are shown. CD8+ / Treg cell ratios were significantly higher in responders vs. non-responders at 8th week of treatment (p=0.00955). Study name: GEN501, subset of 17 patients. [Figure 10E] Figure 1 shows that effector cell proliferation is more effectively inhibited in the presence of CD38+ Tregs compared to CD38- Tregs or negative controls. Error bars represent standard error. Asterisks indicate significant changes. Samples were obtained from multiple healthy donors. Cell proliferation was assessed by dilution of carboxyfluorescein succinimidyl ester (CFSE). [Figure 11] Myeloid-derived suppressor cells (MDSCs) were present in multiple myeloma patients (top graph, boxed cells) and approximately half of the cells expressed CD38 (middle graph, boxed cells). The CD38high MDSC population was depleted in patients treated with a single infusion of DARZALEX™ (daratumumab) (bottom graph, boxed cells). Study name: GEN501, subset of 17 patients. [Figure 12]Figure 1 shows that CD38high MDSC (CD11b+HLADR-CD14-CD33+CD15+) counts were decreased in patients at 1, 4 or 8 weeks of DARZALEX™ (daratumumab) treatment compared to baseline, returning to near baseline at end of treatment (EOT). Patients who relapsed still showed decreased CD38high MDSC. Black squares: non-responders; open squares: patients with at least a minimal response to DARZALEX™ (daratumumab) treatment. Vertical lines indicate median values ​​in each group. Patients 2, 4, 15, 16 and 17 showed initially high CD38high MDSC populations. Study name: GEN501, subset of 17 patients. [Figure 13] Patients with the highest CD38high MDSCs (patients 2, 4, 15, 16 and 17) had the longest progression-free survival (PFS). These patients had either a partial response (PR) or minimal response (MR) to DARZALEX™ (daratumumab) treatment. SD: stable disease; PD: progressive disease. The x-axis shows the PFS for each numbered patient. [Figure 14] This shows that MDSCs are sensitive to DARZALEX™ (daratumumab)-induced ADCC. Daudi cells were used as a positive control of target cells in the assay. The % cell lysis was measured. [Figure 15A] 1 shows that CD38+ Bregs were depleted in patients treated with DARZALEX™ (daratumumab) at weeks 1, 4, and 8 of treatment. [Figure 15B] Figure 2 shows that CD38+ Bregs secrete IL-10 upon stimulation. [Figure 16A]Figure 1 shows antiviral responses as measured by CMV, EBV, and influenza virus-specific (CEF) IFN-γ production in PBMCs from patients treated with DARZALEX™ (daratumumab) who were VGPR during treatment, at baseline, and at the indicated time points. OD: optical density. White bars: negative control; black bars: CEF added; shaded bars: allogeneic PBMCs only. Asterisks indicate statistically significant changes. Pre 4, 8, 10 = 4, 8, or 10 weeks of treatment. [Figure 16B] Figure 1 shows antiviral responses as measured by CMV, EBV, and influenza virus-specific (CEF) IFN-γ production in PBMCs from patients treated with DARZALEX™ (daratumumab) who were in CR during treatment, at baseline, and at the indicated time points. OD: optical density. White bars: negative control; black bars: CEF added; shaded bars: allogeneic PBMCs only. Asterisks indicate statistically significant changes. Pre 4, 8, 10 = 4, 8, or 10 weeks of treatment. [Figure 16C] Figure 1 shows antiviral responses as measured by CMV, EBV, and influenza virus-specific (CEF) IFN-γ production in PBMCs from patients treated with DARZALEX™ (daratumumab) who were in PD during treatment, at baseline, and at the indicated time points. OD: optical density. White bars: negative control; black bars: CEF added; shaded bars: allogeneic PBMCs only. Ns: not significant. Pre 4, 8 = 4 or 8 weeks of treatment. [Figure 16D] Figure 1 shows antiviral responses as measured by CMV, EBV, and influenza virus-specific (CEF) IFN-γ production in PBMCs from patients treated with DARZALEX™ (daratumumab) who were in MR during treatment, at baseline, and at the indicated time points. OD: optical density. White bars: negative control; black bars: CEF added; shaded bars: allogeneic PBMCs only. Ns: not significant. Pre 4, 8 = 4th or 8th week of treatment. [Figure 16E]Shown are the percentages of proliferating virus-reactive T cells in PBMCs from patients treated with DARZALEX™ (daratumumab) who were in VGPR during treatment, at baseline, and at the indicated time points. White bars: negative control; black bars: CEF added. Asterisks indicate statistically significant changes. Pre 4, 8, 10 = 4, 8, or 10 weeks of treatment. [Figure 16F] Shown are the percentages of proliferating virus-reactive T cells in PBMCs from patients treated with DARZALEX™ (daratumumab) who were in CR at baseline and the indicated time points during treatment. White bars: negative control; black bars: CEF added. Asterisks indicate statistically significant changes. Pre 4, 8, 10 = weeks 4, 8, or 10 of treatment. [Figure 17A] 1 shows histograms of FACS analysis showing expression levels of CD38 in natural killer cells (NK), monocytes, B cells and T cells from healthy donors. [Figure 17B] 1 shows histograms of FACS analysis indicating the expression levels of CD38 in plasma cells, natural killer cells (NK), monocytes, B cells, and T cells from multiple myeloma patients. [Figure 17C] Comparison of CD38 mean fluorescence intensity (MFI) in CD38+ Treg, Breg, NK, B cells, and T cells from relapsed and refractory multiple myeloma patients. CD38 was expressed at lower levels in B cells and T cells compared to CD38+ Treg, Breg, and NK cells. [Figure 18] Figure 2 shows that PD-L1 protein is downregulated in PBMC samples from responders (R) and upregulated in non-responders (NR) over time. SD: stable. C1D1: cycle 1, day 1; C3D1, cycle 3, day 1. Y-axis shows log2 protein concentration values. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] As used in this specification and the appended claims, the singular forms "a," "a "n" and "the" refer to plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a cell" includes two or more cells. and the like.

[0020] "CD38" refers to the human CD38 protein (synonym: ADP-ribosyl cyclase 1, c ADPr hydrolase 1, cyclic ADP ribose hydrolase 1). Human CD38 is The amine shown in GenBank accession number NP 001766 and in SEQ ID NO:1 is CD38 has the amino acid sequence 1-21, which represents the cytoplasmic domain, and the transmembrane domain. The amino acid sequence 22-42 represents the domain, and residues 43-44 represent the extracellular domain of CD38. It is known that it is a single-pass type II membrane protein having a .DELTA..times ...

[0021] SEQ ID NO:1 MANCEFSPVSGDKPCCRLSRRAQLCLGVSILVLILVVVL AVVVPRWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRH VDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPC NKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDLTW CGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRFAEA ACDVVHVMLNGSRSKIFDKNSTFGSVEVHNLQPEKVQTLE AWVIHGGREDSRDLCQDPTIKELESIISKRNIQFSCKNIY RPDKFLQCVKNPEDSSCTSEI

[0022] As used herein, "antibody" is intended in a broad sense and includes murine, human, humanized, and and monoclonal antibodies, antibody fragments, bispecific antibodies or multispecific antibodies, including chimeric monoclonal antibodies. Specific antibodies, dimeric, tetrameric or multimeric antibodies, single chain antibodies, domain antibodies, and Any other modified form of an immunoglobulin molecule that contains an antigen-binding site of the required specificity. The compositions also include immunoglobulin molecules.

[0023] Immunoglobulins are divided into five major classes, all of which are based on the heavy chain constant domain amino acid sequence. IgA and IgD can be assigned to IgE, IgG, and IgM. gG is a subset of the isotypes IgA1, IgA2, IgG1, IgG2, IgG3, and IgG The antibody light chains of any vertebrate species are classified into four subtypes: Based on the amino acid sequence, two distinct types are classified: kappa (κ) and lambda (λ). ) can be classified into one of two types.

[0024] An "antibody fragment" refers to a fragment of an antibody that contains a heavy and / or light chain antigen-binding site, e.g., a heavy chain complementarity determining region ( HCDR) 1, 2, and 3, light chain complementarity determining region (LCDR) 1, 2, and 3, heavy chain variable The term "variable domain" refers to a portion of an immunoglobulin molecule that retains the variable domain (VH) or the variable light domain (VL). The antibody fragment is a Fab fragment, which is a monovalent fragment consisting of the VL, VH, CL, and CH1 domains. and two Fab fragments linked by a disulfide bridge at the hinge region. F(ab)2 fragment, which is a fragment of an antibody, Fd fragment consisting of VH and CH1 domains, and An Fv fragment consisting of the VL and VH domains of one arm and a domain consisting of the VH domain Deoxyribonucleic acid (dAb) fragments (Ward et al., Nature 341:544-6, 1989), in which the VH and VL domains are engineered and linked via a synthetic linker. and linked together to form a variety of single chain antibody designs, where VH / V The L domain may pair intramolecularly, or the VH and VL domains may form separate single chains. When expressed by an antibody construct, the molecules are paired together to form single chain Fv (scFv) or These form monovalent antigen-binding sites, such as diabodies. Nos. 98 / 44001, 1988 / 01649, 1994 / 13804, and These antibody fragments can be produced by techniques well known to those skilled in the art. These fragments are obtained using techniques and can be screened for utility in the same manner as full length antibodies. It is cleaned.

[0025] An "isolated antibody" is an antibody or antibodies that are substantially free of other antibodies having different antigenic specificities. (e.g., an isolated antibody that specifically binds CD38 refers to a fragment other than human CD38. However, it is not substantially free of antibodies that specifically bind to human CD38. The isolated antibody binds to Macaca fascicularis (cynomolgus monkey) CD3 8, which may have cross-reactivity to other antigens, such as orthologues of human CD38. In the case of bispecific antibodies, the bispecific antibody specifically binds to two different antigens of interest. The antibody is substantially free of antibodies that specifically bind to antigens other than these two types of target antigens. An isolated antibody may be substantially free of other cellular material and / or chemicals. "Antibody" refers to an antibody isolated with high purity, e.g., 80%, 81%, 82%, 83%, 84%, or 85% purity. %, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94 %, 95%, 96%, 97%, 98%, 99% or 100%.

[0026] "Specific binding" or "specifically binds" or "binds" means an antigen that binds to a substance with a higher degree than other antigens. An antibody that binds to an antigen or an epitope within an antigen with affinity. Typically, an antibody binds to an antigen at equilibrium. Dissociation constant (K D ) is about 1×10 -8 M or less, for example, about 1×10 -9 M or less, approximately 1×10 - 10 M or less, approximately 1×10 -11 M or less, or about 1 x 10 -12 M or less, antigen or intra-antigen Binds to the epitope, typically this K D Non-specific antigens (e.g., BSA, casein) K for binding to D The dissociation constant is at least 100 times smaller than However, antibodies that specifically bind to an antigen or an epitope within an antigen can be used to measure the level of specificity. The body may be, for example, a human or monkey, for example, Macaca fascicularis. monkey, cyno), Pan troglodytes (chimpanzee, chimp) or Callithrix jacchus (common marmoset, marmoset) etc. may have cross-reactivity to the same antigen (homolog) from other species. The body specifically binds to one antigen or epitope, but bispecific antibodies It specifically binds to two different antigens or two different epitopes.

[0027] The antibody variable region consists of a "framework" region separated by three "antigen binding sites". The antigen-binding site is defined using various terms: there are three in the VH (HCDR1, HC HCDR2, HCDR3) and three in VL (LCDR1, LCDR2, LCDR3) Complementarity determining regions (CDRs) are based on sequence variability (Wu and Kabat (1970 )J Exp Med 132:211~50;Kabatet al Sequenc es of Proteins of Immunological Interest ,5th Ed,Public Health Service,National I Institutes of Health, Bethesda, Md., 1991), V There are three "hypervariable" regions in H (H1, H2, and H3) and three in VL (L1, L2, and L3). "HVR" or "HV" refers to the Chothia and Lesk by ia and Lesk (1987) MolBiol 196:901-17 It refers to the regions of an antibody variable domain that are structurally hypervariable as defined. , “IMGT-CDR” (Lefranc et al., (2003) Dev Com parat Immunol 27:55~77) and “specificity-determining residue use” (SDR U)(Almagro(2004)Mol Recognit 17:132~43) International ImMunoGeneTics (IMGT) The database (http: / / www_imgt_org) is a database of standardized numbers for antigen-binding sites. The symbology and definitions are provided. The correspondence between the CDR, HV, and IMGT notations is given in , Lefrancet al., (2003) Dev Comparat Immuno It is described in l 27:55-77.

[0028] As used herein, "Chotia residue" refers to the residue of Al-Lazikani (Al -Lazikani et al.,(1997)J MolBiol 273:927 48).

[0029] "Framework" or "framework sequence" refers to a region defined as an antigen-binding site. The remaining sequence of the variable region, excluding the nucleotide sequence of ... The exact amino acid sequence of the framework can be determined by how the antigen-binding site is defined. It depends on how it is defined.

[0030] A "humanized antibody" refers to an antibody in which the antigen-binding site is derived from a species other than human and the variable region framework A humanized antibody refers to an antibody in which the amino acid sequence within the framework region is derived from a human immunoglobulin sequence. Since the framework may contain substitutions in the expressed human immunoglobulin It is not necessary for the gene to be an exact copy of the clone or germline gene sequence.

[0031] A "human antibody" is an antibody in which both the framework and the antigen-binding sites are derived from sequences of human origin. When the antibody contains a constant region, it is called a constant The regions are also derived from sequences of human origin.

[0032] Human antibodies are antibodies whose variable regions are derived from human germline immunoglobulins or rearranged immunoglobulins. Heavy chain variable regions "derived" from sequences of human origin when obtained from a system using the purine gene Such a system comprises human immunoglobulins displayed on phages, which contain either the light or dark chain variable regions. and transgenic mice carrying human immunoglobulin loci. "Human antibodies" include antibodies derived from, for example, naturally occurring somatic mutations, or for the introduction of targeted substitutions in the framework or antigen-binding site, or both. compared to human germline immunoglobulins or rearranged immunoglobulin genes Typically, a "human antibody" is a human antibody that is a recombinant human antibody. or an amino acid sequence encoded by a rearranged immunoglobulin gene, At least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical. In some cases, a "human antibody" is, e.g., For example, Knappik et al., (2000) J Mol Biol 296:57 The consensus framework obtained from the analysis of human framework sequences described in or, for example, Shi et al., (2010) J MolBiol 397: 385-96 and WO 2009 / 085462, The present invention may include a synthetic HCDR3 integrated into a library of human immunoglobulin genes. .

[0033] Human antibodies derived from human immunoglobulin sequences can be generated by phage display incorporating synthetic CDRs and and / or synthetic frameworks, or other systems to improve antibody properties. can undergo in vitro mutagenesis to produce human antibodies in vivo This results in antibodies that do not naturally occur within the germline repertoire.

[0034] Antibodies in which the antigen binding site is derived from a species other than human are not included in the definition of a human antibody.

[0035] "Recombinant antibodies" refer to antibodies that are derived from transgenic or chromosomal introductions of human immunoglobulin genes. The input animal (e.g., mouse or rat) or a hybridoma prepared therefrom (hereinafter ), an antibody isolated from a host cell transformed to express the antibody. Antibodies isolated from recombinant combinatorial antibody libraries, as well as human immunodeficiency virus (HIV)-associated antibodies By any other means involving splicing globulin gene sequences to other DNA sequences. The antibodies prepared, expressed, generated, or isolated by the present invention or by Fab arm exchange are used in Antibodies produced in vitro, e.g., bispecific antibodies, prepared, expressed, or engineered by recombinant means. The term "antibody" includes all antibodies that are prepared or isolated.

[0036] The term "monoclonal antibody" refers to a preparation of antibody molecules of single molecular composition. A homogeneous antibody composition exhibits a single binding specificity and affinity for a particular epitope and In the case of bispecific monoclonal antibodies, they have dual binding specificities for two distinct epitopes. Thus, a "monoclonal antibody" is one that has been modified by the removal of a C-terminal lysine from the antibody heavy chain. Except for potential known alternatives such as, for example, a single A monoclonal antibody refers to an antibody population with an amino acid composition. Monoclonal antibodies may be monospecific or polyspecific, or monovalent, bivalent, Bivalent antibodies are included in the term monoclonal antibody.

[0037] "Epitope" means the part of an antigen to which an antibody specifically binds. , chemically active (polar, non-polar or hydrophobic, etc.) moieties such as amino acid or polysaccharide side chains. ) surface groups and may have specific three-dimensional structural characteristics and specific charge characteristics. , may consist of adjacent and / or non-adjacent amino acids forming a conformational space unit. For non-contiguous epitopes, amino acids from different parts of the linear sequence of the antigen are Through the folding of protein molecules, they come into close proximity in three-dimensional space.

[0038] A "variant" is a variant of a reference gene by one or more modifications, e.g., substitutions, insertions, or deletions. A polypeptide or polynucleotide that differs from a polypeptide or reference polynucleotide Refers to...

[0039] "In combination with" means that two or more therapeutic agents are administered to a subject together in a mixture. Administered simultaneously as single agents or sequentially in any order as single agents Generally, each drug will be administered at a dose and / or schedule determined for that drug. It is administered at a fixed rate.

[0040] "Treating" or "treatment" refers to a condition in which the purpose is to prevent or reduce an undesirable physiological change or progression of a disease. Slow (reduce) the development or spread of tumors or tumor cells, for example or by providing a beneficial or desirable clinical outcome during treatment. A beneficial or desired clinical outcome is one that is detectable or Relief of symptoms, reduction in the extent of disease, stable disease (i.e., no detectable disease), whether present or undetectable (i.e., no worsening of) the disease state, delay or slowing of disease progression, absence of metastasis, improvement or "Treatment" includes alleviation, palliative care, and remission (whether partial or total). Also, prolonging survival compared to the expected survival if the subject did not receive treatment A subject in need of treatment may be a person suffering from an undesired physiological change or disease. This includes subjects who already have a disease, as well as subjects who are predisposed to having a physiological change or disease. can be.

[0041] A "therapeutically effective amount" refers to a compound, at dosages and for periods of time necessary, to achieve a desired therapeutic result. The therapeutically effective amount refers to an amount that is effective for the treatment of a disease. The therapeutically effective amount may be determined by factors such as the disease state, age, sex, and weight of the individual, and by the ability of a therapeutic agent or combination of therapeutic agents to elicit a desired response in an individual. Examples of indicators of an effective therapeutic agent or combination of therapeutic agents include, for example: For example, improving the patient's health status, reducing tumor burden, stopping or slowing tumor growth, and / or and the absence of metastasis of cancer cells to other locations in the body.

[0042] "Inhibits proliferation" (e.g., with respect to tumor cells) refers to a treatment or combination of treatments that The reduction or slowing of proliferation of tumor cells or tumor tissue in the absence of treatment or therapy compared to the reduction or slowing of proliferation of the same tumor cells or tumor tissue in the absence of treatment or therapy or combination of therapeutic agents in vitro or in vivo It refers to a measurable decrease or delay in the proliferation of tumor cells or tumor tissue. Inhibition of tumor cell or tumor tissue proliferation in vivo is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100% It is possible.

[0043] "Regulatory T cells" or "Tregs" are cells that regulate the activity of other T cells and / or other immune cells. Treg refers to T lymphocytes that normally regulate the immune system by suppressing its activity. + CD4 + CD25 +CD127 dim T cells are fully capable of this phenotype. It is recognized that the host cell may express, but is not limited to, Foxp3.

[0044] "Effector T cells" or "Teff" are involved in tumor cell killing and / or Functions of the immune response, such as activation of antitumor immune responses that can lead to the elimination of tumor cells from the body Teff refers to T lymphocytes that play a key role in + CD4 + or CD8 + may be accompanied by. Teff secretes, contains, or expresses markers such as IFN-γ, granzyme B, and ICOS. It is recognized that Teffs may not be entirely restricted to these phenotypes. do.

[0045] "Treg function" or "Treg function" refers to the regulation of the host immune response and / or the inhibition of autoimmunity. The function of Treg is to suppress the immune response to CD8 + T cells, natural NK cells,

[0046]

number

[0047] "Inhibition of Treg function" or "inhibition of Treg function" refers to the in vivo administration of Tregs to animals or human subjects. Measured in vitro or in vivo by conventional techniques known in the art. The level of Treg function that can be detected by Tregs is decreased. For example, at least about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, The decrease in Treg function may be 70%, 80%, 90%, 95%, 99% or 100%. "Inhibition" includes, for example, inhibition via antibody effector functions such as antibody-dependent cellular cytotoxicity (ADCC). These include reducing the number of Tregs by killing them with immunization.

[0048] "Myeloid-derived suppressor cells" or "MDSCs" are hematopoietic cells that are involved in the development of macrophages / The monocyte marker CD11b and the granulocyte marker Gr-1 / Ly-6G were The phenotype of MDSCs refers to a specialized cell population that expresses CD11b + HLA -DR - CD14 - CD33 + CD15 + MDSCs can be classified as mature antigen-presenting cells. The expression of the markers MHC class II and F480 is low or undetectable. DSCs are immature cells of the myeloid lineage and include macrophages, neutrophils, dendritic cells, monocytes, or granulocytes. MDSCs are found in normal bone marrow of adults and animals, and can further differentiate into several cell types, including erythrocytes. or may be found naturally at normal hematopoietic sites such as the spleen.

[0049] "Inhibition of MDSC function" or "inhibition of MDSC function" refers to the in vivo Measured in vitro or in vivo by conventional techniques known in the art. This refers to a decreased level of MDSC function that can be achieved by For example, at least about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, The function of MDSCs may be reduced by 70%, 80%, 90%, 95%, 99% or 100%. Inhibition can involve, for example, killing MDSCs via antibody effector functions such as ADCC. These include reducing the number of MDSCs by inhibiting reactive oxygen species (ROS), peroxygenases, and oxidative stress. Nitrite generation, improved arginase metabolism due to high levels of arginase, and Various mechanisms, such as increased nitric oxide synthase, may lead to proliferation, clonal expansion, or cyclin synthesis. MDSCs can suppress T cell responses, such as IFN-γ and IL-4 production. IFN-γ is a nitric oxide-dependent cytokine that stimulates the immune system to produce inflammatory cytokines. It can activate MDSCs by inducing the activity of NOS2. Th2 cytokines such as leukin-4 (IL-4) and IL-13 inhibit arginase- It can activate MDSCs, which can lead to the induction of NOS2 or AR Metabolism of L-arginine by either G1 or G2 may result in inhibition of T cell proliferation. The activity of both enzymes is thought to mediate T cell apoptosis through the production of reactive nitric oxide species. This may cause.

[0050] "Treg-associated disease" refers to a disease or disorder associated with T regulatory cells (Treg). Treg-associated diseases may involve a loss of Treg function, e.g., suppression of anti-tumor responses or inhibition of effector T The Treg-mediated disease may be cancer. " and "Treg-mediated disease" are used interchangeably herein.

[0051] "Enhanced effector T cell responses" or "enhanced T cell responses" refers to the administration of Maintain or enhance biological functions in vitro or in vivo. or as an effector for regenerating or reactivating exhausted or inactive T cells. This refers to the enhancement or stimulation of T-cells. Typical T-cell responses include proliferation, production of gamma interferon, and CD8 + This enhancement can be measured by T cell secretion, antigen responsiveness, or clonal expansion. Methods are known to those skilled in the art.

[0052] "MDSC-associated disease" refers to a disease or condition associated with myeloid-derived suppressor cells (MDSCs). MDSC-associated diseases refer to disorders that impair MDSC function, such as anti-tumor response or effector function. The MDSC-mediated disease may be cancer. The terms "disease" and "MDSC-mediated disease" are used interchangeably herein.

[0053] "Regulatory B cells" or "Bregs" refer to B lymphocytes that suppress immune responses. g, CD19 + CD24 + CD38 + cells and IL secreted by Bregs It may suppress immune responses by inhibiting Br-10-mediated T cell proliferation. There exists a subset of these, e.g., Ding et al., (2015) Human I It is recognized that this is described in Immunology 76:615-621.

[0054] "Breg-associated disease" refers to a disease or disorder associated with regulatory B cells. The disease may result, for example, from Breg-mediated suppression of anti-tumor responses or effector T cell proliferation. The Breg-mediated disease may be cancer. The terms "disease" and "disease" are used interchangeably herein.

[0055] A "patient" includes any human or non-human animal. A "non-human animal" includes any vertebrate animal. , e.g., non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles "Patient" and "subject" are used interchangeably herein. will be done.

[0056] The present invention specifically binds CD38 regardless of whether or not the tumor cell expresses CD38. The present invention further provides a method of treating a patient with a solid tumor using an antibody that inhibits the Regulatory T cells (Treg), myeloid-derived suppressor cells (MDSC) or regulatory B cells (B The present invention further provides a method for treating a patient having a Treg-mediated disease. Regulating MDSC or Breg activity and CD38 positivity and / or high levels of these immunosuppressants Methods for treating solid tumors associated with sex cells are provided.

[0057] The present invention is based, at least in part, on the anti-CD38 antibody DARZALEX™ (Darzalex). Tumumab) has immunomodulatory activity in patients and inhibits immune suppressive Tregs, MDSCs and reduce Breg counts and CD8 + T cell count and CD8 + Increase the ratio of Tregs to Se, CD8 + Promotes the formation of central memory cells and increases T cell clonality. It is based on the discovery that

[0058] DARZALEX™ (daratumumab) and other anti-CD38 antibodies are CD38 positivity due to antibody effector functions such as DC, ACDP and apoptosis Its cell elimination ability makes it an ideal candidate for the treatment of hematological malignancies and plasma cell disorders such as multiple myeloma. The efficacy of these drugs is being evaluated in clinical trials for their immunomodulatory activity in promoting adaptive immune responses. Other immunomodulatory antibodies (anti-PD1, anti-CTLA4) have not been shown to induce antitumor responses. They work by targeting components of the immune system that they suppress. For example, anti-PD1 antibodies , increase T cell proliferation, stimulate antigen-specific memory responses, and It has been shown to partially relieve Treg-mediated suppression of effector T cells (e.g. (See, for example, U.S. Patent No. 8,779,105.) Two types of anti-PD-L1 antibodies are available for the treatment of melanoma. -1 antibody, OPDIVO® (nivolumab) and KEYTRUDA® (pembrolizumab) are currently approved, and these antibodies have been used to treat a variety of solid tumors, e.g. Non-small cell lung cancer, prostate, head and neck, gastrointestinal, stomach, prostate, fallopian tube, ovarian, pancreatic, breast and brain cancer, It is currently in clinical development for kidney, bladder, urethral, ​​esophageal and colorectal cancer. Its new drug, YERVOY® (ipilimumab), is approved for the treatment of melanoma. YERVOY® (ipilimumab) and another anti-CTLA-4 antibody, thrombin timepoint Remelimumab is also being used in the treatment of prostate, non-small cell lung, ovarian, gastrointestinal, gastric, colorectal, renal, esophageal, and It is in development for genitourinary cancers.

[0059] Without wishing to be bound by any particular theory, it is believed that the DARZA Based on the immunomodulatory effects observed with LEX™ (daratumumab), DAR ZALEX™ (daratumumab) and other anti-CD38 antibodies are effective in treating solid tumors This may be the case in patients treated with DARZALEX™ (daratumumab). Due to the general activation of immune responses, patients with CD38-negative solid tumors may be able to tolerate anti-C They may respond similarly to D38 antibody therapy.

[0060] The present invention relates to a method for treating solid tumors by administering a therapeutically effective amount of an antibody that specifically binds CD38 to a patient. Treating a patient with a solid tumor, including administering to a patient in need thereof for a sufficient period of time. The present invention provides a method for

[0061] The present invention also relates to a method for treating a patient with a disease characterized by the initiation of treatment with a therapeutically effective amount of an antibody that specifically binds CD38. to a patient in need thereof for a period of time sufficient to treat a Treg-mediated disease. Also provided are methods of treating patients with Treg-mediated disorders, including Treg-associated disorders.

[0062] The present invention also relates to a method for treating bone marrow-derived suppositories comprising administering to the bone marrow a therapeutically effective amount of an antibody that specifically binds CD38. Administered to patients in need for a period of time sufficient to treat lesser cell (MDSC)-mediated diseases Also provided is a method of treating a patient having an MDSC-mediated disease comprising administering to the patient a therapeutically effective amount of the ...

[0063] The present invention also provides a method for administering a therapeutically effective amount of an antibody that specifically binds CD38 to a regulatory B cell. This includes administering it to patients in need of it for a period of time sufficient to treat Breg-mediated diseases. Also provided are methods of treating patients with Breg-mediated diseases, including

[0064] The present invention also relates to a method for the treatment of neurodegenerative diseases by contacting regulatory T cells (Treg) with an antibody that specifically binds CD38. Also provided is a method for suppressing the activity of regulatory T cells, comprising:

[0065] The present invention also relates to a method for treating myeloid-derived suppressor cells (MDSCs) that specifically bind CD38. Also provided is a method of inhibiting the activity of MDSCs comprising contacting with the antibody.

[0066] The present invention also relates to a method for the treatment of inflammatory bowel disease by contacting regulatory B cells (Bregs) with an antibody that specifically binds CD38. Also provided is a method for inhibiting Breg activity, comprising:

[0067] The present invention also provides a method for the treatment of a patient's inflammatory bowel disease by administering to the patient an antibody that specifically binds CD38. and reducing the number of regulatory T cells (Tregs) in patients with solid tumors. Methods of treatment are also provided.

[0068] The present invention also provides a method for the treatment of a patient's inflammatory bowel disease by administering to the patient an antibody that specifically binds CD38. and reducing myeloid-derived suppressor cell (MDSC) numbers in patients with solid tumors. Also provided are methods of treating a patient having

[0069] The present invention also provides a method for the treatment of a patient's inflammatory bowel disease by administering to the patient an antibody that specifically binds CD38. and reducing the number of regulatory B cells (Bregs) in patients with solid tumors. Methods of treatment are also provided.

[0070] The present invention also provides a method for administering an antibody that specifically binds CD38 to a patient for a period of time sufficient to enhance an immune response. Also provided is a method of enhancing an immune response in a patient comprising administering to a patient in need thereof.

[0071] In some embodiments, the patient has a viral infection.

[0072] The present invention also provides a method for administering an antibody that specifically binds CD38 to a patient for a period of time sufficient to treat a viral infection. Also provided is a method of treating a viral infection in a patient, comprising administering the same to a patient in need thereof. Provide.

[0073] In some embodiments, the immune response is an effector T cell (Teff) response.

[0074] In some embodiments, the Teff response is CD4 +T cells or CD8 + T cells It is mediated through.

[0075] In some embodiments, the Teff response is CD4 + Mediated by T cells.

[0076] In some embodiments, the Teff response is CD8 + Mediated by T cells.

[0077] In some embodiments, the Teff response is CD8 + Increased T cell count, CD8 + T cells Increased proliferation, increased T cell clonal expansion, CD8 + Increased memory cell formation, antigen-dependent Increased antibody production, or increased cytokine, chemokine, or interleukin production It is.

[0078] T cell proliferation can be measured, for example, by measuring the rate of DNA synthesis using tritiated thymidine. or measurement of in vitro production of interferon-γ (IFN-γ) or By measuring the absolute number or percentage of T cells in a cell population derived from a patient sample using known methods. This can be evaluated as follows.

[0079] Clonal expansion can be determined, for example, by sequencing TCRs from a pool of T cells using known methods. This can be evaluated as follows.

[0080] Memory cell generation can be achieved by, for example, isolating naive T cells (CD45RO - / C D62L + ) memory T cells (CD45RO + / CD62L high ) as a percentage It can be evaluated by measuring.

[0081] Cytokine, chemokine or interleukin production, e.g., interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), IL-1, IL-2, IL-3, I L-4, IL-6, IL-8, IL-10, IL-12, IL-13, IL-16, IL The production of E-18, IL-23, MIP-1α, MIP-1β, RANTES, and CCL4 was This can be assessed using standard methods such as LISA or ELLISPOT assays.

[0082] Antigen-specific antibody production is measured from patient samples using ELISA or radioimmunoassay. It can be evaluated using standard methods such as radioimmunoassay (RIA).

[0083] The meaning of "increasing" or "increasing" various Teff responses is readily understood. In a test sample or subject, as compared to a control, e.g., treated with an anti-CD38 antibody, In patients who received anti-CD38 antibody therapy, the response was compared to that of the same patients before treatment, or in a patient or group of patients compared with a patient or group of patients who were non-responsive to the same treatment At least about 5%, at least about 10%, 25%, 50%, 51%, 52%, 53%, %, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63% %, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73% %, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83% %, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% %, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 110%, 120% , 130%, 140%, 150%, 200%, 250%, 300%, 350%, 400% or more. Typically, the increase is statistically significant.

[0084] Similarly, the numbers of Tregs, MDSCs and / or Bregs are "decreased" or "reduced" The meaning of "reduction" or "reducing" is readily understood. or in subjects, e.g., in patients treated with an anti-CD38 antibody, compared to a control. When compared with the same patient before treatment, or in patients who responded to anti-CD38 antibody treatment or In a patient group, at least Approximately 10%, 25%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57% %, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67 %, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77 %, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87 %, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% %, 98%, 99%, 100%, 110%, 120%, 130%, 140%, 150%, There may be a decrease of 200%, 250%, 300%, 350%, 400% or more. Typically, this reduction is statistically significant.

[0085] In some embodiments, an antibody that specifically binds CD38 is an immune suppressor cell. Inhibits the function of.

[0086] In some embodiments, the immune suppressor cells are regulatory T cells (Tregs), bone marrow These are myeloblast-derived suppressor cells (MDSC) or regulatory B cells (Breg).

[0087] In some embodiments, Tregs are CD3 + CD4 + CD25 + CD127 dim T cells.

[0088] In some embodiments, CD3 + CD4 + CD25 + CD127 dim The cells are Expresses xp3.

[0089] In some embodiments, CD3 + CD4 + CD25 + CD127 dim T cells are It expresses CD38.

[0090] Treg function, such as the suppressive ability of Teff cells, is e.g. The inhibitory effect of a Teff cell on T cell proliferation in a multiple receptor tyrosine kinase (MLR) response can be assessed using known methods.

[0091] Treg function can be measured, for example, by measuring the relative number of Tregs compared to Teff (e.g., CD8 + / Increase in the Treg cell ratio) can be determined by measuring Treg or a subpopulation of Treg, e.g., CD38 + Treg This can be inhibited by reducing the production of β-lactamase by direct killing.

[0092] In some embodiments, Treg function is inhibited by killing of Treg cells.

[0093] In some embodiments, killing of Tregs is mediated by antibodies that specifically bind CD38. Antibody-induced antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis induced by antibodies These may be mediated by complement-dependent cytotoxicity (ADCP), complement-dependent cytotoxicity (CDC) or apoptosis.

[0094] In some embodiments, the killing of Tregs is mediated by ADCC.

[0095] In some embodiments, CD38 + Tregs are killed.

[0096] In some embodiments, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58% or 60% Tregs are killed.

[0097] CD38 is expressed only on a subset of Tregs and MDSCs, making it a promising candidate for treatment of solid tumor patients. Treatment with this agent does not result in systemic depletion of Tregs and MDSCs, potentially improving safety. It is expected.

[0098] In some embodiments, the MDSCs are CD11b + HLA-DR - CD14 - CD3 3 + CD15 + It is a cell.

[0099] In some embodiments, CD11b + HLA-DR - CD14 - CD33 + CD15 + MDSCs express CD38.

[0100] MDSC function can be inhibited, for example, by reducing MDSC numbers through direct cell killing. can be inhibited.

[0101] In some embodiments, MDSC function is determined by CD38 + Inhibited by killing of MDSCs can be.

[0102] In some embodiments, the killing of MDSCs is by an antibody that specifically binds CD38. Induced by antibody-induced antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis ( The induction of cytotoxicity is mediated by complement-dependent cytotoxicity (ADCP), complement-dependent cytotoxicity (CDC) or apoptosis.

[0103] In some embodiments, the killing of MDSCs is mediated by ADCC.

[0104] In some embodiments, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58% or 60% of MDSCs are killed.

[0105] In some embodiments, Bregs are CD19 + CD24 + CD38 + It is a cell.

[0106] Breg function can be reduced by, for example, directly killing Bregs, thereby decreasing their numbers. Thus, it can be inhibited.

[0107] In some embodiments, Breg function is determined by CD38 + Inhibited by killing of Breg can be.

[0108] In some embodiments, killing of Bregs is by an antibody that specifically binds CD38. Induced by antibody-induced antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis ( The induction of cytotoxicity is mediated by complement-dependent cytotoxicity (ADCP), complement-dependent cytotoxicity (CDC) or apoptosis.

[0109] In some embodiments, Breg killing is mediated by ADCC.

[0110] In some embodiments, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58% or 60% Breg is killed.

[0111] Tregs play an important role in maintaining peripheral self-tolerance. 4 + CD25 hi Tregs are produced in the thymus and are essential for establishing the identity and suppressive function of the Treg lineage. Tregs express Foxp3, a transcription factor required for the establishment and maintenance of immune cells. For example, they can accumulate in tumor tissues (e.g., within tumors) where they suppress the effector functions of tumor antigen-specific T cells. This leads to insufficient antitumor responses. + Treg Increased density is associated with poor prognosis in various solid tumors, including pancreatic, ovarian, and hepatocellular carcinoma. In mouse models, depletion of Tregs improved antitumor immunity and tumor rejection. However, it can also lead to the development of autoimmune diseases.

[0112] Myeloid-derived suppressor cells (MDSCs) are early myeloid progenitors at various stages of differentiation. These are heterogeneous populations of cells, immature granulocytes, macrophages, and dendritic cells. It accumulates in large amounts in patients with chronic kidney disease and has a strong immunosuppressive function, inhibiting natural killer (NK) and nasal lymphocytes. Cytotoxic activity of local killer T cells (NKT) and CD8 + Mediated by T cells The mechanism by which NK cells are inhibited is currently poorly understood. However, it increases the production of arginase 1 / ARG1 and nitric oxide synthase 2 (NOS2) Multiple pathways, including regulation of T cell proliferation and regulating IL-1 expression, are responsible for MDSC-mediated T cell suppression. NOS1 and NOS2 metabolize L-arginine and together or separately inhibit the translation of the T cell CD3 ζ chain. In addition, MDSCs suppress immune responses, inhibit T cell proliferation, and promote T cell apoptosis. It secretes immunosuppressive cytokines and induces the development of regulatory T cells.

[0113] MDSCs are induced by inflammatory cytokines and are abundant in infectious and inflammatory conditions. These accumulate in the blood, bone marrow, and secondary lymphoid organs of tumor-bearing mice. Their presence in the tumor microenvironment may play a causative role in promoting tumor-associated immunosuppression. It is said to fulfill this.

[0114] MDSC includes colon cancer, melanoma, hepatocellular carcinoma, head and neck squamous cell carcinoma, non-small cell lung cancer, and renal cell carcinoma. , has been described in patients with pancreatic adenocarcinoma and breast cancer (Mandruzzato et al. l.,(2009)J Immunol 182:6562~6568;Liu et al. al.,(2009)J Cancer Res Clin Oncol 136:35 ~45;Ko et al.,(2009)Clin Cancer Res 15:2 148~2157;Morse et al.,(2009)Expert OpinB iol Ther 9:331~339;Diaz-Montero et al.,( 2009) Cancer Immunol Immunother 58:49~59; Corzo et al.,(2009)J Immunol 182:5693~57 In cancer patients, Diaz-Montero et al. 009) Cancer Immunol Immunother 58:49~59) , have proposed that accumulation of MDSCs correlates with further disease progression and poor prognosis.

[0115] Tumor-infiltrating Bregs have been identified in solid tumors and act through a variety of mechanisms. , e.g., CD8 + By suppressing the antitumor activity of T cells and NK cells, may promote cell proliferation and metastasis (see, e.g., Ding et al., (2015) Human (Described in Immunology 76:615-62).

[0116] “Antibody-dependent cytotoxicity”, “antibody-dependent cell-mediated cytotoxicity”, or “ADCC” Binding of antibody-coated target cells via Fc gamma receptors (FcγR) expressed on effector cells It has lytic activity against cells such as natural killer cells, monocytes, macrophages, and neutrophils. It is a mechanism for inducing cell death that depends on interaction with effector cells that are involved in the cell death process. For example, NK cells express FcγRIIIa, whereas monocytes express FcγRI, FcγRII, and FcγRIIIa expression. Killing of antibody-coated target cells, such as CD38-expressing cells, occurs via membrane pores. As a result of effector cell activity through the secretion of morphogenetic proteins and proteases To assess the ADCC activity of antibodies that specifically bind CD38, the antibodies were CD38 expressing cells may be added in combination with effector cells, which may act as antigen-antibody. When activated by the cytoplasmic complex, it can result in cytolysis of the target cell. Labeling (e.g., radioactive substrates, fluorescent dyes, or natural intracellular proteins) from lysed cells is usually performed. Effector cells for such assays include peripheral blood cells. Representative target cells include peripheral blood mononuclear cells (PBMCs) and NK cells. In an exemplary assay, the target cells include Tregs or MDSCs expressing , 20 μCurie 51 The cells were labeled with Cr for 2 hours and washed extensively. The cell concentration of target cells was 1 x 10 6 cells / mL, but anti-CD38 antibodies vary The assay was performed with effector:target cells at a ratio of 40:1. After 3 hours of incubation at 37°C, the assay was From lysed cells arrested by isolation 51 The release of Cr was measured in a scintillation counter. The percentage of cytotoxicity is measured by adding 3% perchloric acid to the target cells. The lysis can be calculated as the percentage of maximum lysis that can be induced by

[0117] "Antibody-dependent cellular phagocytosis" ("ADCP") refers to the process by which the phagocytosis of cells, e.g., macrophages or dendritic cells, This indicates the mechanism of killing of antibody-coated target cells through internalization by phagocytes such as vesicles. ADCPs are genetically engineered to express GFP or other tagged molecules. This can be evaluated by using CD38-expressing Tregs or MDSCs as target cells. The effector to target cell ratio can be, for example, 4:1. The effector cells can be Incubation with target cells for 4 hours with or without anti-CD38 antibody After incubation, the cells can be detached using Accutase. Macrophages were identified using fluorescently labeled anti-CD11b and anti-CD14 antibodies. Percent phagocytosis can be determined using standard methods, e.g., by measuring CD11 + and CD14 + It can be determined based on the percent GFP fluorescence in the macrophages.

[0118] "Complement dependent cytotoxicity" or "CDC" is a process that inhibits the binding of target-binding antibodies to Fc effector domains. binds to and activates complement component C1q, which then activates the complement cascade. Complement refers to a mechanism for inducing cell death that activates complement, resulting in the death of the target cell. Activation of complement can also result in the deposition of complement components on the target cell surface, which in turn activates leukocytes. Binding of complement receptors (eg, CR3) to the antibody facilitates ADCC.

[0119] The ability of monoclonal antibodies to induce ADCC is dependent on their oligosaccharide moieties. Human IgG1 or IgG3 can be enhanced by genetic engineering. It is N-glycosylated at sn297, where the majority of the glycans are N-glycosylated with the known dimers. It is in the form of branched G0, G0F, G1, G1F, G2, or G2F. Antibodies produced by CHO cells that do not have glycan fragments typically have at least about 85% glycan fragments. Core fucose content from biantennary complex oligosaccharides attached to the Fc region. Removal of FcγRIIIa resulted in improved FcγRIIIa binding without altering antigen binding or CDC activity. Such mAbs enhance ADCC of antibodies through binding to the osmolarity of the medium. (Konno et al.,(2012)Cytotechnology 64:24 9-65), and application of the mutant CHO line Lec13 as a host cell line (Shields et al. et al., (2002) J Biol Chem 277:26733~26740) , application of the mutant CHO line EB66 as a host cell line (Olivier et al., (2010)MAbs 2(4),Epub ahead of print;PMID 20562582), and its suitability as a host cell line for the rat hybridoma cell line YB2 / 0. (Shinkawa et al., (2003) J Biol Chem 278: 3466-3473), α1,6-fucosyltransferase (FUT8) gene Introduction of specific small interfering RNA (Mori et al., (2004) Biol. Echnol Bioeng 88:901-908) or β-1,4-N-acetylglucosamine glucosaminyltransferase III and Golgi α-mannosidase II or Co-expression of kifunensine, a potent α-mannosidase I inhibitor (Ferrara et al. t al.,(2006)J Biol Chem 281:5032~5036;Fe rrara et al.,(2006)Biotechnol Bioeng 93: 851~861;Xhou et al.,(2008)Biotechnol Bio eng 99:652-65) and other defucosylated Fc oligosaccharides with biantennary complex structure. This has been achieved using a variety of methods that have been reported to result in relatively high expression of cloning antibodies. In the method of the present invention, and in the numbered embodiments listed below, A1 induced by anti-CD38 antibodies used in some embodiments DCC may also be enhanced by certain substitutions in the antibody Fc. The substitution can be performed, for example, by inserting a nucleotide at amino acid position 25 as described in U.S. Pat. No. 6,737,056. 6, 290, 298, 312, 356, 330, 333, 334, 360, 378, or The substitution is at 430 (residue numbering according to the EU index).

[0120] In some embodiments, an antibody that specifically binds CD38 comprises a substitution in the antibody Fc. Includes.

[0121] In some embodiments, an antibody that specifically binds CD38 comprises 256, 260, 270, 280, 290, 300, 310, 320, 335, 340, 356, 360, 370, 380, 390, 395, 400, 410, 420, 430, 440, 450, 46 290, 298, 312, 356, 330, 333, 334, 360, 378 or 430 (Residue numbering is according to the EU index).

[0122] In some embodiments, the antibody that specifically binds CD38 has a fucose content of about 0% ~About 15%, e.g. 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7 %, 6%, 5%, 4%, 3%, 2%, 1% or 0% of the biantennary glycan structures .

[0123] In some embodiments, an antibody that specifically binds CD38 has a fucose content of about 50 %, 40%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 14%, 13 %, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or 0% have biantennary glycan structures.

[0124] Substitutions and reduced fucose content in Fc allow the ADC of antibodies that specifically bind CD38. C activity can be enhanced.

[0125] "Fucose content" refers to the amount of fucose monosaccharide in the glycan at Asn297 The relative amount of fucose is the ratio of fucose-containing structures to the total number of sugar structures. The construction can be carried out in a number of ways, for example: 1) as described in WO 2008 / 077546; Next, MALDI-TOF of the N-glycosidase F treated sample (e.g., complex, hybridization) was performed. 2) the use of enzymatic release of the Asn297 glycan; extraction, followed by derivatization and HPLC with fluorescence detection (UPLC) and / or HPLC- 3) detection / quantification by MS (UPLC-MS) between the first and second GlcNAc monosaccharides by Endo S or other enzymes that cleave and attach a fucose to the first GlcNAc Immunoprecipitation of native or reduced mAb with or without processing of the glycan at Asn297. 4) Enzymatic digestion (e.g., trypsin or endopeptide digestion) The mAb was digested into its component peptides by the enzyme Lys-C, followed by HPLC-MS (UPL C-MC) or 5) PNGase F at Asn297. Deglycosylation of mAb oligosaccharides to mAb proteins through specific enzymatic deglycosylation using The released oligosaccharides can be characterized and quantified by separation from the fluorophore. The glycans are labeled with carboxyl groups and then subjected to a variety of complementary techniques that allow for detailed characterization of the glycan structures. These can be separated and identified by comparing the experimental masses with the theoretical masses. Matrix-assisted laser desorption / ionization (MALDI) mass spectrometry and ion-exchange HPLC ( Determination of the degree of sialylation by normal phase HPLC (GlycoSep C) Separation and quantification of oligosaccharide types according to hydrophilicity criteria using high performance capillary Separation and quantification of oligosaccharides by HPCE-LIF do.

[0126] As used herein, "low fucose" or "low fucose content" refers to an antibody that has about 0% It refers to having a fucose content of ~15%.

[0127] As used herein, "normal fucose" or "normal fucose content" refers to More than about 50% of the antibody is present, usually more than about 60%, 70%, 80%, or more than 85%. It refers to having fucose content.

[0128] In some embodiments, antibodies that specifically bind CD38 inhibit apoptosis. , can induce killing of Tregs, MDSCs and / or Bregs. Methods for this purpose are well known and include, for example, staining with Annexin IV using standard methods. The anti-CD38 antibody used in the method of the present invention can inhibit approximately 20%, 25%, 30%, or 40% of the cells. %, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80 %, 85%, 90%, 95%, or 100% apoptosis can be induced.

[0129] In some embodiments, Teff or immune suppressor cells are present in the bone marrow or peripheral blood. exist.

[0130] In some embodiments, the Teff or immune suppressor cells are present in the bone marrow.

[0131] In some embodiments, the Teff or immune suppressor cells are present in peripheral blood. .

[0132] In some embodiments, an antibody that specifically binds CD38 is + T cell Tr Increase the ratio to eg.

[0133] In some embodiments, an antibody that specifically binds CD38 is + Central Molly cell CD8 + Increases the ratio of CD8 to naive cells. + Central Memory The cells are CD45RO + / CD62L +high These cells can be identified as CD8 + Nai The CD45RO / CD62L + Can be identified as a cell.

[0134] In some embodiments, the antibody that specifically binds CD38 is a non-agonistic antibody. do.

[0135] Nonagonistic antibodies that specifically bind CD38 have in v Proliferation of peripheral blood mononuclear cell samples in vitro was compared with that of isotype control antibodies or medium alone. The term "antibody" refers to an antibody that does not induce proliferation significantly compared to the proliferation induced by the antibody.

[0136] In some embodiments, non-agonistic antibodies that specifically bind CD38 are statistically Induces peripheral blood mononuclear cell (PBMC) proliferation in a non-significant manner. PBMC proliferation is PBMCs were isolated from healthy donors and incubated in 200 μL of RPMI in the presence or absence of the test antibody. 1 x 10 cells in a flat-bottom 96-well plate 5 By culturing cells / well After incubation at 37°C for 4 days, 30 μL of 3 H-thymidine (16. 7 μCi / mL) and incubation may be continued overnight. 3 H-thymidine incorporation is Packard Cobra Gamma Counter (Packard Instruments) The data can be evaluated using a Meriden, DT, USA) according to the manufacturer's instructions. It can be calculated as the mean cpm (± SEM) of PBMC obtained from several donors. Statistical significance or non-significance between samples cultured in the presence or absence of the antibody was determined using standard methods. Calculate using the method.

[0137] An exemplary anti-CD38 antibody that can be used in the methods of the present invention is DARZALEX™ (DARZALEX®). DARZALEX™ (daratumumab) is represented by the sequence ID No. 4 and 5, the amino acid sequences of the heavy chain variable region (VH) and the light chain variable region (VL), Heavy chain complementarity determining region 1 (HCDR1), HCDR2 and H CDR3, and light chain complementarity determining region 1 (LCDR) of SEQ ID NOs: 9, 10 and 11, respectively 1) LCDR2 and LCDR3 are included, and daratumumab is of the IgG1 / κ subtype. and is described in U.S. Patent No. 7,829,693. DARZALEX™ (Daratumumab) The amino acid sequence of the heavy chain is shown in SEQ ID NO: 12 and the amino acid sequence of the light chain is , as shown in SEQ ID NO:13.

[0138] In some embodiments, an antibody that specifically binds CD38 has a heavy chain variable domain of SEQ ID NO:4. and an antibody comprising a variable domain (VH) and a light chain variable domain (VL) of SEQ ID NO:5, which binds to CD38. compete regarding.

[0139] In some embodiments, the antibody that specifically binds CD38 is human CD38 (SEQ ID NO: No. 1) the region SKRNIQFSCKNIYR (SEQ ID NO: 2) and the region EKVQTLEA It binds at least to WVIHGG (SEQ ID NO:3).

[0140] SEQ ID NO:1 MANCEFSPVSGDKPCCRLSRRAQLCLGVSILVLILVVVL AVVVPRWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRH VDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPC NKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDLTW CGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRFAEA ACDVVHVMLNGSRSKIFDKNSTFGSVEVHNLQPEKVQTLE AWVIHGGREDSRDLCQDPTIKELESIISKRNIQFSCKNIY RPDKFLQCVKNPEDSSCTSEI

[0141] SEQ ID NO:2 SKRNIQFSCKNIYR

[0142] SEQ ID NO:3 EKVQTLEAWVIHGG

[0143] SEQ ID NO:4 EVQLLESGGGLVQPGGSLRLSCAVSGFTFNSFAMSWVRQ APGKGLEWVSAISGSGGGTYYADSVKGRFTISRDNSKNTL YLQMNSLRAEDTAVYFCAKDKILWFGEPVFDYWGQGTLVT VSS

[0144] SEQ ID NO:5 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQK PGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLE PEDFAVYYCQQRSNWPPTFGQGTKVEIK

[0145] SEQ ID NO:6 SFAMS

[0146] SEQ ID NO:7 AISGSGGGTYYADSVKG

[0147] SEQ ID NO:8 DKILWFGEPVFDY

[0148] SEQ ID NO:9 RASQSVSSYLA

[0149] SEQ ID NO:10 DASNRAT

[0150] SEQ ID NO:11 QQRSNWPPTF

[0151] SEQ ID NO:12 EVQLLESGGGLVQPGGSLRLSCAVSGFTFNSFAMSWVRQ APGKGLEWVSAISGSGGGTYYADSVKGRFTISRDNSKNTL YLQMNSLRAEDTAVYFCAKDKILWFGEPVFDYWGQGTLVT VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG TQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEL LGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPS REEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGK

[0152] SEQ ID NO:13 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQK PGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLE PEDFAVYYCQQRSNWPPTFGQGTKVEIKRTVAAPSVFIFP PSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC

[0153] The antibody was identified as having a binding affinity to CD38 using well-known in vitro methods. and DARZALEX™ (daratumumab) having a VH of SEQ ID NO:5 and a VL of SEQ ID NO:6. Competition with a reference antibody can be assessed. In a typical method, CD38 is recombinantly expressed CHO cells were incubated with unlabeled reference antibody for 15 min at 4°C, and then incubated with excess May be incubated with fluorescently labeled test antibody for 45 minutes at 4°C in PBS / BSA After washing with , fluorescence can be measured by flow cytometry using standard methods. In another exemplary method, the extracellular portion of human CD38 is attached to the surface of an ELISA plate. An excess of unlabeled reference antibody can be added for about 15 minutes, followed by biotin. After washing in PBS / Tween, the biotinylated test antibody can be added. Antibody binding was confirmed by horseradish peroxidase (HRP)-conjugated streptavidin and standard In a competitive assay, the signal detected by the control can be detected using a conventional method. It is readily apparent that the control antibody may be labeled and the test antibody unlabeled. The reference antibody inhibits the binding of the test antibody or the test antibody inhibits the binding of the reference antibody to CD38. At least 80% of the cases, for example, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 9 ...100%, 101%, 102%, 103%, 1 7%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 9 A test antibody competes with a reference antibody when it inhibits by 7%, 98%, 99%, or 100%. The epitope of the test antibody can be determined, for example, by peptide mapping or using known methods. The peptide may be further defined by hydrogen / deuterium protection assays using NMR or by crystal structure determination.

[0154] The region SKRNIQFSCKNIYR (SEQ ID NO: 2) of human CD38 (SEQ ID NO: 1) and Antibodies that bind to the region EKVQTLEAWVIHGG (SEQ ID NO: 3) can be isolated, for example, using standard methods and and those described herein, mice were bred to have the amino acid sequences shown in SEQ ID NOs: 2 and 3. and then is immunized with a peptide having the sequence and the peptide is identified using, for example, ELISA or mutagenesis studies. The antibody can be purified by characterizing the antibodies obtained for binding to the tide.

[0155] The present invention also relates to the region SKRNIQFSCKNIYR (sequence number 1) of human CD38. Lane number 2) and anti-CD38 binding to the region EKVQTLEAWVIHGG (SEQ ID NO: 3) Treating a patient with a solid tumor, comprising administering the antibody to a patient in need thereof. The epitope of the antibody used in the method of the invention is SEQ ID NO: 2 or SEQ ID NO: In some embodiments, the antibody comprises some or all of the residues having the sequence shown in SEQ ID NO:3. The epitope is a region of human CD38 (SEQ ID NO: 1) called SKRNIQFSCKNIYR (sequence At least one amino acid in the region EKVQTLEAWVIHGG (sequence no. 2) and at least one amino acid in sequence number 3). The epitope is a region of human CD38 (SEQ ID NO: 1) called SKRNIQFSCKNIYR (sequence At least two amino acids in the region EKVQTLEAWVIHGG (sequence no. 2) and at least two amino acids in sequence number 3). The epitope is a region of human CD38 (SEQ ID NO: 1) called SKRNIQFSCKNIYR (sequence At least three amino acids in the region EKVQTLEAWVIHGG (sequence no. 2) and at least three amino acids in sequence number 3).

[0156] In some embodiments, an antibody that specifically binds CD38 is selected from SEQ ID NOs: 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 7 and 8, including the amino acid sequences of HCDR1, HCDR2 and HCDR3.

[0157] In some embodiments, antibodies that specifically bind CD38 are those having SEQ ID NO:9, 10 and 11, comprising the amino acid sequences of LCDR1, LCDR2 and LCDR3.

[0158] In some embodiments, an antibody that specifically binds CD38 is selected from SEQ ID NOs: 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 7, 8, 9, 10 and 11 HCDR1, HCDR2, HCDR3, LCDR1, LCD It includes the amino acid sequences of LCDR2 and LCDR3.

[0159] In some embodiments, an antibody that specifically binds CD38 has a sequence similar to SEQ ID NO: 4, 95% VH that is 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 5 and SEQ ID NO: 95 %, 96%, 97%, 98%, 99% or 100% identical to the VL.

[0160] In some embodiments, an antibody that specifically binds CD38 has the VH of SEQ ID NO:4 and Contains the VL of SEQ ID NO:5.

[0161] In some embodiments, an antibody that specifically binds CD38 has a heavy chain of SEQ ID NO: 12 and and a light chain of SEQ ID NO:13.

[0162] Other exemplary anti-CD38 antibodies that can be used in any embodiment of the invention are: .

[0163] VH of SEQ ID NOs: 14 and 15, respectively, as described in U.S. Pat. No. 7,829,693 The VH and VL of mAb003 are identical to those of IgG1 / κ. It can be expressed as:

[0164] SEQ ID NO:14 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAFSWVRQ APGQGLEWMGRVIPFLGIANSAQKFQGRVTITADKSTSTA YMDLSSLRSEDTAVYYCARDDIAALGPFDYWGQGTLVTVS SAS

[0165] SEQ ID NO:15 DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQK PEKAPKSLIYAASSLQSGVPSRFSGSGGSGTDFTLTISSLQ PEDFATYYCQQYNSYPRTFGQGTKVEIK

[0166] VH of SEQ ID NOs: 16 and 17, respectively, as described in U.S. Pat. No. 7,829,693 The VH and VL of mAb024 are identical to those of IgG1 / κ. It can be expressed as:

[0167] SEQ ID NO:16 EVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQ MPGKGLEWMGIIYPHSDARYSPSFQGQVTFSADKSISTA YLQWSSLKASDTAMYYCARHVGWGSRYWYFDLWGRGTLVT VSS

[0168] SEQ ID NO:17 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQK PGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLE P EDFAVYYCQQRSNWPPTFGQGTKVEIK;

[0169] VH of SEQ ID NOs: 18 and 19, respectively, as described in U.S. Pat. No. 8,088,896 MOR-202 (MOR-03087) including the VH and VL sequences. and VL may be expressed as IgG1 / κ.

[0170] SEQ ID NO:18 QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYYMNWVRQ APGKGLEWVSGISGDPSNTYYADSVKGRFTISRDNSKNTL YLQMNSLRAEDTAVYYCARDLPLVYTGFAYWGQGTLVTVS S

[0171] SEQ ID NO:19 DIELTQPPSVSVAPGQTARISCSGDNLRHYYVYWYQQKP GQAPVLVIYGDSKRPSGIPERFSGSNSGNTATLTISGTQA EDEADYYCQTYTGGASLVFGGGGTKLTVLGQ

[0172] VH of SEQ ID NOs: 20 and 21, respectively, as described in U.S. Pat. No. 8,153,765 Isatuximab, including the VH and VL sequences. It can be expressed as κ.

[0173] SEQ ID NO:20: QVQLVQSGAEVAKPGTSVKLSCKASGYTFTDYWMQWVKQ RPGQGLEWIGTIYPGDGDTGYAQKFQGKATLTADKSSKTV YMHLSSLASEDSAVYYCARGDYYGSNSLDYWGQGTSVTVS S

[0174] SEQ ID NO:21 DIVMTQSHLSMSSTSLGDPVSITCKASQDVSTVVAWYQQK PGQSPRRLIYSASYRYIGVPDRFTGSGAGTDFTFTISSVQ AEDLAVYYCQQHYSPPYTFGGGTKLEIK

[0175] Other exemplary anti-CD38 antibodies that can be used in the methods of the invention include: International Publication Nos. 05 / 103083, 06 / 125640, and 07 / 042309 No. 08 / 047242 or No. 14 / 178820 Some examples include:

[0176] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 will be delivered to patients in need for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor comprising administering

[0177] The present invention also includes a therapeutically effective amount of a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15. The antibody that specifically binds CD38 is administered to a patient in need thereof for a period of time sufficient for the treatment of solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to a subject a therapeutically effective amount of a compound comprising administering to said subject a therapeutically effective amount of a compound comprising a compound selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,

[0178] The present invention also includes a therapeutically effective amount of a VH of SEQ ID NO: 16 and a VL of SEQ ID NO: 17. The antibody that specifically binds CD38 is administered to a patient in need thereof for a period of time sufficient for the treatment of solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to a subject a therapeutically effective amount of a compound comprising administering to said subject a therapeutically effective amount of a compound comprising a compound selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,

[0179] The present invention also includes a therapeutically effective amount of a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19. The antibody that specifically binds CD38 is administered to a patient in need thereof for a period of time sufficient for the treatment of solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to a subject a therapeutically effective amount of a compound comprising administering to said subject a therapeutically effective amount of a compound comprising a compound selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,

[0180] The present invention also includes a therapeutically effective amount of a VH of SEQ ID NO:20 and a VL of SEQ ID NO:21. The antibody that specifically binds CD38 is administered to a patient in need thereof for a period of time sufficient for the treatment of solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to a subject a therapeutically effective amount of a compound comprising administering to said subject a therapeutically effective amount of a compound comprising a compound selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,

[0181] In some embodiments, the solid tumor is a melanoma.

[0182] In some embodiments, the solid tumor is lung cancer.

[0183] In some embodiments, the solid tumor is non-small cell lung cancer (NSCLC).

[0184] In some embodiments, the solid tumor is non-squamous NSCLC.

[0185] In some embodiments, the solid tumor is a lung adenocarcinoma.

[0186] In some embodiments, the solid tumor is renal cell carcinoma (RCC) (e.g., renal clear cell carcinoma or renal papillary cell carcinoma) or its metastasis.

[0187] In some embodiments, the solid tumor is mesothelioma.

[0188] In some embodiments, the solid tumor is nasopharyngeal carcinoma (NPC).

[0189] In some embodiments, the solid tumor is colorectal cancer.

[0190] In some embodiments, the solid tumor is prostate cancer or castration-resistant prostate cancer.

[0191] In some embodiments, the solid tumor is stomach cancer.

[0192] In some embodiments, the solid tumor is ovarian cancer.

[0193] In some embodiments, the solid tumor is gastric cancer.

[0194] In some embodiments, the solid tumor is hepatocellular carcinoma.

[0195] In some embodiments, the solid tumor is pancreatic cancer.

[0196] In some embodiments, the solid tumor is thyroid cancer.

[0197] In some embodiments, the solid tumor is squamous cell carcinoma of the head and neck.

[0198] In some embodiments, the solid tumor is a carcinoma of the esophagus or gastrointestinal tract.

[0199] In some embodiments, the solid tumor is breast cancer.

[0200] In some embodiments, the solid tumor is fallopian tube cancer.

[0201] In some embodiments, the solid tumor is brain cancer.

[0202] In some embodiments, the solid tumor is a urethral cancer.

[0203] In some embodiments, the solid tumor is a genitourinary cancer.

[0204] In some embodiments, the solid tumor is endometriosis.

[0205] In some embodiments, the solid tumor is cervical cancer.

[0206] In some embodiments, the solid tumor is a metastasis of a cancer.

[0207] In some embodiments, the solid tumor lacks detectable CD38 expression.

[0208] Using well-known methods, compare expression to a control, e.g., expression detected with an anti-CD38 antibody versus an isoform The expression of the antibody in solid tumor tissue or isolated from solid tumors was compared to that detected by a control antibody of the same type. Solid tumors lack detectable CD3 expression when CD38 expression on the transplanted cells is not statistically significant. 8 expression is absent.

[0209] The anti-CD38 antibodies used in the methods of the present invention may be derived, for example, from a phage display library. Alternatively, the phage may be selected to encode a human immunoglobulin or a portion thereof (e.g., F ab, single chain antibody (scFv), or unpaired or paired antibody variable regions) It has been genetically engineered to express the Mol Biol 296:57~86;Krebs et al.,(2001)J Immunol Meth 254:67~84;Vaughan et al.,(1 996)Nature Biotechnology 14:309~314;Shee ts et al.,(1998) PITAS(USA)95:6157~6162;H oogenboom and Winter,(1991)J Mol Biol 22 7:381;Marks et al.,(1991)J Mol Biol 222: 581). The CD38-binding variable domain is described, for example, in Shi et al., (2010) J Mol Biol 397:385-96 and International Publication No. 09 / 085462 The antibody is loaded onto the phage as a fusion protein with the bacteriophage pIX coat protein. The antibody library may be isolated from a phage display library expressing the variable regions of the light and dark chains. The antibodies were screened for binding to the human CD38 extracellular domain, and the positive clones obtained The clones were further characterized, and Fabs were isolated from clone lysates and subsequently synthesized as full-length antibodies. Such phage display methods for isolating human antibodies are well known in the art. See, e.g., U.S. Pat. Nos. 5,223,409 and 5,403, , No. 484, No. 5,571,698, No. 5,427,908, No. 5,580, No. 717, No. 5,969,108, No. 6,172,197, No. 5,885,7 No. 93, No. 6,521,404, No. 6,544,731, No. 6,555,31 See US Pat. Nos. 3, 6,582,915, and 6,593,081.

[0210] In some embodiments, the anti-CD38 antibody is an IgG1, IgG2, IgG3, or I It is the gG4 isotype.

[0211] The Fc portion of antibodies is responsible for antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent cell phagocytosis. These mediate the effector functions of antibodies, such as complement-dependent cytotoxicity (ADCP) or complement-dependent cytotoxicity (CDC). Such a function may mediate the phagocytic activity of the FC effector domain(s). by binding to Fc receptors on immune cells that have cytotoxic or lytic activity, or by Fc effector This can be mediated by binding of the complement domain(s) to components of the complement system. The effect(s) mediated by the complement component or the complement component may be expressed by target cells, e.g., CD38 expression. The human IgG isotypes, IgG1, I IgG2, IgG3, and IgG4 exhibit unique capabilities with regard to effector function. ADCC is mediated by IgG1 and IgG3, whereas ADCP is mediated by IgG1, IgG2 , IgG3, and IgG4, whereas CDC is mediated by IgG1 and IgG3. It can be mediated.

[0212] An antibody that is substantially identical to an antibody comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5 is disclosed in the present invention. As used herein, the term "substantially identical" means The amino acid sequences of the two antibodies VH or VL to be compared are identical or have only slight similarities. A negligible difference means that the antibody has a specific function that does not adversely affect the antibody's properties. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 1 in the antibody heavy or light chain The percent identity can be calculated using, for example, Vector or A of NTI v.9.0.0 (Invitrogen, Carlsbad, CA) This was determined by pairwise alignment using the default settings of the lignX module. The protein sequences of the present invention can be used as query sequences to search public or patent databases. A search against a database may be performed to identify, for example, related sequences. An exemplary program used to run the program is XBLAST, using the default settings. Or use the BLASTP program (http: / / www.ncbi.nlm / nih. gov), or GenomeQuest™ (GenomeQuest, Westb orough, MA) suite, which is performed against antibodies that specifically bind CD38. Exemplary substitutions that may be made include, for example, substitutions with amino acids having similar charge, hydrophobicity, or stereochemical properties. Conservative substitutions are also used to improve antibody properties, such as stability or affinity. It may also be performed to enhance the activity of antibodies or to improve antibody effector function. 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids Acid substitutions can be made, for example, to the heavy or light chain of the anti-CD38 antibody. Scanning mutagenesis has been previously described (MacLenna n et al.,Acta Physiol.Scand.Suppl.643:55 ~67,1998;Sasaki et al.,Adv.Biophys.35:1~ 24, 1998), any naturally occurring residue in VH or VL can also be substituted with alanine. Desired amino acid substitutions can be determined by one of skill in the art at the time such substitutions are desired. Acid substitutions can be performed, for example, by PCR mutagenesis (U.S. Patent No. 4,683,195). Libraries of mutants can be generated using well-known methods, e.g., random codon (NNK) or non-random codons, such as the 11 amino acids (Ala, Cys, Asp, DV coding for Glu, Gly, Lys, Asn, Arg, Ser, Tyr, and Trp K codon and screen the library for mutants with the desired properties. The generated mutants may be generated by binding to CD38 in vitro. binding to, or inducing ADCC, ADCP, or apoptosis, or CD38 enzymatic activity These can be tested for their ability to modulate the Cut.

[0213] In some embodiments, antibodies that specifically bind CD38 have broad affinity (K D ) In one embodiment according to the present invention, the numbering scheme listed below can be used to bind human CD38. In some of the various embodiments described above, the present invention may be implemented by those skilled in the art. CD38 specific as determined by surface plasmon resonance or Kinexa assays Antibodies that bind heterologously with high affinity, e.g., K D About 10 -7 M or less, for example, these but is not limited to 1 to 9.9 (i.e. 1, 2, 3, 4, 5, 6, 7, 8, or 9, etc.) (any range or value within that range) x 10 -8 M, 10 -9 M, 10 -10 M, 10 -1 1 M, 10 -12 M, 10 -13 M, 10 -14 M, 10 -15 M, or any range Binds to CD38 at or near the range of affinities. An example of an affinity is 1 × 10 -8 M or less Another example of affinity is 1×10 -9 It is below M.

[0214] In some embodiments, the antibody that specifically binds CD38 is a bispecific antibody. The VL and / or VH regions of an existing anti-CD38 antibody or those described herein may be used as a The newly identified VL and VH regions were then genetically engineered into bispecific full-length antibodies. Such bispecific antibodies may be used as disclosed in U.S. Pat. No. 7,695,936, International Publication No. No. 04 / 111233, U.S. Patent Application Publication No. 2010 / 0015133, U.S. Patent No. Publication No. 2007 / 0287170, International Publication No. 2008 / 119353, U.S. Pat. Publication No. 2009 / 0182127, U.S. Patent Application Publication No. 2010 / 0286374 No. 2011 / 0123532, and WO 2011 / 13174. No. 6, International Publication No. 2011 / 143545, or U.S. Patent Application Publication No. 2012 / 01 To form bispecific antibodies, monospecific antibodies can be synthesized using techniques such as those described in US Pat. No. 4,9876. The antibody of the present invention can be produced by adjusting the CH3 interaction between the heavy chains of the heterologous antibodies. Additional bispecific structures into which the VL and / or VH regions can be incorporated include, for example, bivariate domain immunoglobulin (WO 2009 / 134776) or leucine The antibody has two arms with specificity, such as a collagen zipper or collagen dimerization domain. Structures containing various dimerization domains for binding (WO 2012 / 022811 , U.S. Patent No. 5,932,448, and U.S. Patent No. 6,833,441).

[0215] For example, bispecific antibodies can be produced according to the methods described in WO 2011 / 131746. In vitro, in a cell-free environment, the C Asymmetric mutations were introduced into the H3 region, and the disulfide bonds were isomerized under reducing conditions. by forming a bispecific heterodimeric antibody from two parent monospecific homodimeric antibodies. In this method, a first monospecific bivalent antibody (e.g., anti-CD41, 38 antibody) and a second monospecific bivalent antibody, which binds the CH3 domain to promote heterodimer stability. These antibodies are genetically engineered to have specific substitutions in the hinge region. Under reducing conditions sufficient for cysteines in the These are then incubated together, which results in the generation of bispecific antibodies via Fab arm exchange. Incubation conditions may optimally be returned to non-reducing conditions. Exemplary reducing agents include 2-mercaptoethylamine (2-MEA), dithiothreitol, (DTT), dithioerythritol (DTE), glutathione, tris(2-carboxylate) Triethyl)phosphine (TCEP), L-cysteine, and beta-mercaptoethanol , preferably 2-mercaptoethylamine, dithiothreitol, and tris(2-mercaptoethylamine). For example, the reducing agent is selected from the group consisting of at least At a temperature of 20° C., in the presence of at least 25 mM 2-MEA or at least 0. In the presence of 5 mM dithiothreitol, pH 5 to 8, for example pH 7.0 or pH 7. In 4, an incubation of at least 90 minutes may be used.

[0216] Exemplary CH3 mutations that can be used in the first and second heavy chains of a bispecific antibody include: K409R and / or F405L.

[0217] The methods of the present invention can be used to treat any category of animal subject. Examples of such animals include mammals such as humans, rodents, dogs, cats, and livestock. can be.

[0218] Administration / Pharmaceutical Compositions In the method of the present invention, the antibody that specifically binds CD38 is The antibody may be provided in a suitable pharmaceutical composition comprising an antibody that binds to the antibody and a pharma- ceutically acceptable carrier. A carrier is a diluent, adjuvant, or excipient that is administered with an antibody that specifically binds CD38. Such vehicles may be peanut oil, soybean oil, mineral oil, sesame oil, etc. Liquids such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as oils. For example, 0.4% saline and 0.3% glycine may be used. The fluids are sterile and generally free of particulate matter. They are sterilized using conventional and well known sterilization techniques (e.g. The composition can be sterilized by, for example, filtration. Pharmaceutically acceptable auxiliary substances required for the preparation of the composition, such as pH adjusting and buffering agents, stabilizers, The formulation may contain thickeners, lubricants, colorants, etc. The concentration of the antibody that specifically binds 8 may vary widely, i.e., from about 0.5% by weight. from less than about 1% by weight, usually up to about 15% or 20% by weight, up to about 25% by weight, up to 30%, 35%, 40%, 45%, or 50% by weight; Also, depending on the particular method of administration selected, the required dose, fluid volume, viscosity, etc. Suitable vehicles and formulations (other human proteins, e.g. human serum) are selected based on: (including albumin) is described, for example, in Remington: The Science and Practice of Pharmacy, 21 st Edition, Troy, DBed., Lipincott Williams and Wilkins, P. hiladelphia,PA 2006,Part 5,Pharmaceutica Manufacturing pp 691-1092, especially pp.95 Please refer to 8~989.

[0219] The method of administering an antibody that specifically binds CD38 in the method of the present invention can be performed using methods known in the art. As is well known in the art, parenteral administration, for example, intradermal, intramuscular, intraperitoneal, intravenous or subcutaneous, , intrapulmonary, transmucosal (oral, intranasal, intravaginal, rectal), or other means understood by those of skill in the art. Antibodies that specifically bind CD38 can be isolated using known methods. The drug may be administered intratumorally to lymph node drainage sites for localized delivery within the tumor.

[0220] Antibodies that specifically bind CD38 can be administered, for example, by intravenous (iv) infusion or bolus injection. Parenterally, by injection, by any suitable route, such as intramuscularly, subcutaneously, or intraperitoneally, Intravenous infusion can be administered for example, 15, 30, 60, 90, 120, 18 0, or 240 minutes, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 This may be carried out for 1 or 12 hours.

[0221] The dose administered to the patient is sufficient to alleviate or at least partially halt the disease being treated. (a "therapeutically effective amount"), and sometimes from 0.005 mg to about 100 mg / kg. kg, for example, about 0.05 mg to about 30 mg / kg, or about 5 mg to about 25 mg / kg, or about 4 mg / kg, about 8 mg / kg, about 16 mg / kg or about 24 mg / kg, or may be, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 mg / kg. However, even higher amounts, for example about 15, 16, 17, 18, 19, 20, 21, 22, 23 , 24, 25, 30, 40, 50, 60, 70, 80, 90 or 100 mg / kg It's fine.

[0222] For example, fixed unit doses of 50, 100, 200, 500, or 1000 mg may be given. or based on the surface area of ​​the patient, e.g., 500, 400, 300, 250, 200, or 100 mg / m 2 Usually, 1 to 8 doses (e.g., 1 , 2, 3, 4, 5, 6, 7, or 8) may be administered, but 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or higher doses may be administered. It is possible.

[0223] In the method of the present invention, the administration of an antibody that specifically binds CD38 is performed for one day, two days, three days, or 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 5 weeks, 6 weeks, 7 weeks, 2 months Repeat the course of treatment after 3, 4, 5, 6 or more months. Repeated administration is possible, as is long-term administration. For example, the CD38 specific antibody in the methods of the present invention may be administered at the same or different doses. The antibody binding to for 8 weeks, followed by 8 mg / kg or 16 mg / kg every 2 weeks for an additional 16 weeks. Then, patients were administered 8 mg / kg or 16 mg / kg every 4 weeks. It can be done.

[0224] In the method of the present invention, an antibody that specifically binds CD38 is administered, for example, once a week for six months. The above may be administered as a maintenance therapy.

[0225] For example, the antibody that specifically binds CD38 in the method of the present invention may be 24, 12, 8, Using single or divided doses every 6, 4, or 2 hours, or a combination of these , as a daily dose in an amount of about 0.1 to 100 mg / kg, e.g., 0.5, 0. 9, 1.0, 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 1 3, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 , 27, 28, 29, 30, 40, 45, 50, 60, 70, 80, 90 or 100 mg / kg, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 , 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or on at least one of the 40 days, or on the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th , 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, or 20th week For at least one week of the week, or any combination thereof, the method may be provided.

[0226] The antibody that specifically binds CD38 in the method of the present invention also reduces the risk of cancer progression. delaying the onset of events in the progression of cancer and / or causing cancer remission It may also be administered prophylactically to reduce the risk of recurrence. This may be due to other biological factors. In patients where it is difficult to identify the location of a tumor known to be present due to may be particularly useful.

[0227] The antibodies that specifically bind CD38 in the methods of the present invention can be lyophilized for storage and use. The protein can be reconstituted in a suitable carrier prior to use. This technique is not applicable to conventional protein preparations. It has been shown to be effective and can be prepared using well-known freeze-drying and reconstitution techniques. do.

[0228] In the methods of the invention, the antibody that specifically binds CD38 is administered in combination with a second therapeutic agent. It may be given.

[0229] In the methods of the present invention, an antibody that specifically binds CD38 is administered in the presence of a chemotherapeutic agent known to those of skill in the art. or other anti-cancer therapies. Compounds useful in the treatment of cancer, including antiproliferative or other cytotoxic agents, alkylating agents, substituted agents, Metabolite antagonists, anti-microtubule inhibitors, topoisomerase inhibitors, receptor tyrosine kinase inhibitors Examples of chemotherapy drugs include thiotepa and cyclophosphamide. Alkylating agents, such as cyclosphosphamide (CYTOXAN®); Alkyl sulfonates such as sulfane, improsulfane and piposulfane; benzo Benzodopa, Carboquone, Meturedopa, and Uredopa Aziridines such as altretamine, triethylenemelamine, triethylenephosphoramide (trietylenephosphoramide), triethylenethiophosphoramide (triethylenethiophos Ethyleneimine, including phaoramide and trimethylolmelamine and methylamelamines; chlorambucil, chlornaphazine, Chlorophosphamide, estramustine, ifosfamide, meclo Retamine, mechlorethamine oxide hydrochloride, melphalan, nobuenbikin, fenestrone Nitrogen such as phosphorus, prednimustine, trofosfamide, and uracil mustard Mustard; Carmustine, Chlorozotocin, Fotemustine, Lomustine, Nimustine, Nitrosoureas such as ranimustine; aclacinomysins, actinomycins mycin, anthramycin, azaserine, bleomycin, cactino calicheamicin, carabicin, carminomycin, carzino Filin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-di Azo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, Idarubicin, Marcelomycin, Mitomycin, Mycophenolic acid, Nogalamycin , olivomycin, peplomycin, porfiromycin, puromycin Isin, Querramycin, Rodrubicin, Streptonigrin, Streptozocin, Tuberculin Antibiotics such as lucidin, ubenimex, zinostatin, zorubicin; methotrexate and antimetabolites such as 5-FU; denopterin, methotrexate, pteropterin, Folic acid analogs such as rimetrexate; fludarabine, 6-mercaptopurine, thiamperidone Purine analogs such as phosphatidylcholine and thioguanine; ancitabine, azacitidine, 6-azaurylamide Zin, Carmofur, Cytarabine, Dideoxyuridine, Doxifluridine, Enocitabin pyrimidine analogs such as floxuridine and thrombin; calsterone and dromos propionate androgens such as tanolone, epithiostanol, mepitiostane, and testolactone; Anti-adrenal drugs such as aminoglutethimide, mitotane, and trilostane; folinic acid folic acid supplements such as aceglatone, aldophosphamide glycosides, aminolevulinic acid nicotinic acid; amsacrine; bestrabsil; bisantrene; edatrexate ); defofamine; demecolcine; diaziquone; eflornithine fornithine); elliptinium acetate; etoglucide; gallium nitrate; hydroxyurea; Intinane;Lonidamine;Mitoguazone;Mitoxantrone;Mopidamol;Nitracrine ;Pentostatin;Phenamet;Pirarubicin;Podophyllic acid;2-Ethylhydrazine do; procarbazine; PSK (registered trademark); razoxane; schizophyllan; spirogermani 2,2',2”-Trichlorotriethylamine;Um;Tenuazonic acid;Triazicon;2,2',2”-Trichlorotriethylamine;Um Retan; Vindesine; Dacarbazine; Mannomustine; Mitobronitrol; Mitolactone ru; pipobroman; gacytosine; arabinoside ("Ara-C"); Rofosfamide; thiotepa; paclitaxel (TAXOL®), docetaxel Taxoids or taxanes, such as (TAXOTERE®) and their analogs family;Chlorambucil;Gemcitabine;6-Thioguanine;Mercaptopyridine methotrexate; platinum analogs such as cisplatin and carboplatin; vincristine; stine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxin Santron; Vincristine; Vinorelbine; Navelbine; Novantrone; Teniposide; Daunomycin; Aminopterin; Xeloda; Ibandronate; CPT-11; Topoisomerase Retinolase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids esperamicin; capecitabine; NEXAVAR® (sorafenib), SUTENT® (sunitinib), VOTRIENT™ (pazopanib), PALLADIA™ (torceranib), ZACTIMA™ (vandetanib), RECENTIN® (cediranib), regorafenib (BAY 73-450 6), axitinib (AG013736), lestaurtinib (CEP-701), TA RCEVA® (erlotinib), IRESSA™ (gefitinib), G ilotrif® (afatinib), TYKERB® (lapatinib) ), receptor tyrosine kinase and / or angiogenesis inhibitors, including neratinib, and Examples of compounds that may be used include pharma- ceutically acceptable salts, acids, or derivatives of any of the above. Also included within the definition are antibodies that act to regulate or block the action of hormones in tumors. Hormonal drugs, such as tamoxifen, raloxifene, and aromatase inhibitors )-imidazole, 4-hydroxytamoxifen, trioxyphene, ketoxyphene , LY 117018, onapristone, and FARESTON® (Tremifu) antiestrogens such as flutamide, nilutamide, bicalutamide, antiandrogens, such as bromoprolide, and goserelin; and pharma- ceutically acceptable salts thereof; acid, or a derivative of any of the above. Wiemann et al., 198 5, Medical Oncology (Calabresi et aL, eds.) , Chapter 10, McMillan Publishing Any other conventional cytotoxic compound is applicable to the method of the present invention.

[0230] Representative antibodies that can be used in combination with antibodies that specifically bind CD38 in the methods of the present invention The drugs include tyrosine kinase inhibitors, as well as IRESSA (gefitinib) and and Tarceva® (erlotinib), as well as other HER2-related targeted therapies. Representative examples of HER2 include antagonists of HER2, HER3, HER4, or VEGF. Antagonists include CP-724-714, HERCEPTIN (Trademark) (pertuzumab), OMNITARG™ (pertuzumab), TAK-165, TYKERB (Lapatinib) (EGFR and HER2 inhibitor), and GW-28297 Representative HER3 antagonists include anti-Her3 antibodies (e.g., Representative HER4 antigens include those described in Japanese Patent Application Publication No. 2004 / 0197332. As antagonists, anti-HER4 siRNA (e.g., Maatta et al., Mol Biol Cell 17:67-79, 2006) are representative. A useful VEGF antagonist is Avastin™ (bevacizumab).

[0231] Representative antibodies that can be used in combination with antibodies that specifically bind CD38 in the methods of the present invention Drugs may include standard of care treatments for solid tumors or immune checkpoint inhibitors.

[0232] The second therapeutic agent in the methods of the invention may be an immune checkpoint inhibitor.

[0233] In some embodiments, the immune checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD- L1 antibody, anti-PD-L2 antibody, anti-LAG3 antibody, anti-TIM3 antibody, or anti-CTLA-4 antibody It is a body.

[0234] In some embodiments, the immune checkpoint inhibitor is an antagonist anti-PD-1 Antibody, antagonist anti-PD-L1 antibody, antagonist anti-PD-L2 antibody, antagonist The antibody is an antagonistic anti-LAG3 antibody, or an antagonistic anti-TIM3 antibody.

[0235] In some embodiments, the immune checkpoint inhibitor is an anti-PD-1 antibody.

[0236] In some embodiments, the immune checkpoint inhibitor is an anti-PD-L1 antibody.

[0237] In some embodiments, the immune checkpoint inhibitor is an anti-PD-L2 antibody.

[0238] In some embodiments, the immune checkpoint inhibitor is an anti-LAG3 antibody.

[0239] In some embodiments, the immune checkpoint inhibitor is an anti-TIM3 antibody.

[0240] In some embodiments, the immune checkpoint inhibitor is an anti-CTLA-4 antibody.

[0241] Any antagonist anti-PD-1 antibody may be used in the methods of the invention. Representative anti-PD-1 antibodies include OPVIDO® (nivolumab) and KEYTRUD A® (pembrolizumab). OPVIDO® (nivolumab) is described, for example, in U.S. Pat. No. 8,008,449 (antibody 5C4) and is represented by SEQ ID NO: 24 and VL of SEQ ID NO: 25. Zumab) is described, for example, in U.S. Pat. No. 8,354,509 and has the structure of SEQ ID NO: 22. The VH and VL of SEQ ID NO: 23. The amino acid sequences of nivolumab and pembrolizumab are also , available through the CAS Registry. Additional PD-1 antibodies that can be used are described in U.S. Pat. ,332,582, U.S. Patent Application Publication No. 2014 / 0044738, International Publication No. No. 14 / 17966 and U.S. Patent Application Publication No. 2014 / 0356363. do.

[0242] An "antagonist" is an agent that, when bound to a cellular protein, inhibits the natural ligand of that protein. It refers to a molecule that inhibits at least one reaction or activity induced by at least one A small number of responses or activities that are inhibited in the absence of an antagonist (e.g., a negative control) At least about 30%, 40%, 45%, 50% greater than at least one reaction or activity , 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, young is 100% inhibited or the inhibition is compared to the inhibition in the absence of antagonist. If the difference is statistically significant, the molecule is an antagonist. Antagonists include antibodies, The ligand may be a soluble ligand, a small molecule, DNA or RNA such as siRNA. Typical responses or activities induced by PD-1 include those induced by its receptor PD-L1 or PD -L2 binds antigen-specific CD4 + or CD8 + Cell proliferation may be decreased, or is that interferon-γ (IFN-γ) production by T cells may be decreased, e.g. For example, it suppresses the immune response to tumors. The response or activity is mediated by binding to its receptor, e.g., galectin-9, and is mediated by antigen-specific CD4 + or CD8 + Cell proliferation may be decreased, IFN-γ production by T cells may be decreased, and surface expression of CD137 (CD4 + or CD8 + The fact that the amount of cellular This results in, for example, suppression of the immune response to tumors. Specific for antagonist PD-1, antagonist PD-L2, and TIM-3 Binding antagonist antibodies induce an immune response by inhibiting the inhibitory pathway.

[0243] SEQ ID NO:22 QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQ APGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTA YMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVS S

[0244] SEQ ID NO:23 EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHW YQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDFTLTI SSLEPEDFAVYYCQHSRDLPLTFGGGGTKVEIK

[0245] SEQ ID NO:24 QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQ APGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTL FLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSS

[0246] SEQ ID NO:25 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQK PGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLE PEDFAVYYCQQSSNWPRTFGQGTKVEIK

[0247] Anti-PD-L1 antibodies that enhance the immune response may be used in the methods of the invention (e.g., The representative anti-PD-L1 antibody that can be used is Durbarma. and, for example, U.S. Patent Application Publication No. 2009 / 003666. No. 055944, U.S. Pat. Nos. 8,552,154, 8,217,149, and The invention is described in US Pat. No. 8,779,108.

[0248] Durvalumab comprises a VH of SEQ ID NO:26 and a VL of SEQ ID NO:27.

[0249] Atezolizumab comprises a VH of SEQ ID NO:28 and a VL of SEQ ID NO:29.

[0250] Avelumab comprises a VH of SEQ ID NO:30 and a VL of SEQ ID NO:31.

[0251] SEQ ID NO:26 EVQLVESGGG LVQPGGSLRLSCAASGFTFSRYWMSWVR QAPGKGLEWVAN IKQDGSEKYYVDSVKGRFTISRDNAKN SLYLQMNSLRAEDTAVYYCAREG GWFGELAFDYWGQGTL VTVSS

[0252] SEQ ID NO:27 EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQ K PGQAPRLLIYDASSRATGIPDRFSGSGSGTDFTLTISR LEPEDFAVYYCQQYGSLPWTFGQGTKVEIK

[0253] SEQ ID NO:28 EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQ APGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTA YLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS

[0254] SEQ ID NO:29 DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQK PGKAPKLLIYSASFLYSGVPSRFSGSGSGTFTLTISSLQ PEDFATYYCQQYLYHPATFGQGTKVEIK

[0255] SEQ ID NO:30 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYIMMWVRQ APGKGLEWVSSIYPSGGITFYADTVKGRFTISRDNSKNTL YLQMNSLRAEDTAVYYCARIKLGTVTTVDYWGQGTLVTVS S

[0256] SEQ ID NO:31 QSALTQPASVGSGSPGQSITISCTGTSSDVGGYNYVSWYQ QHPGKAPKLMIYDVSNRPSGVSNRFSGSKSGNTASLTISG LQAEDEADYYCSSYTSSSTRRVFGTGTKVTVL

[0257] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P Administered in combination with D-1 antibody to patients in need thereof for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising:

[0258] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P Administered in combination with D-1 antibody to patients in need thereof for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising:

[0259] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P Administered in combination with the D-L1 antibody to patients in need thereof for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to the patient a therapeutically effective amount of a therapeutically effective amount of a compound selected from the group consisting of ribozyme, cyclosporine ...

[0260] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 is an anti-P Administered in combination with the D-L1 antibody to patients in need thereof for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to the patient a therapeutically effective amount of a therapeutically effective amount of a compound selected from the group consisting of ribozyme, cyclosporine ...

[0261] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P Administered in combination with the D-L1 antibody to patients in need thereof for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to the patient a therapeutically effective amount of a therapeutically effective amount of a compound selected from the group consisting of ribozyme, cyclosporine ...

[0262] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P Administered in combination with the D-1 antibody to patients in need thereof for a period of time sufficient to enhance the immune response Also provided is a method of enhancing an immune response in a patient, comprising:

[0263] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P Administered in combination with the D-1 antibody to patients in need thereof for a period of time sufficient to enhance the immune response Also provided is a method of enhancing an immune response in a patient, comprising:

[0264] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P It should be administered to a patient in need thereof in combination with the D-L1 antibody for a period of time sufficient to enhance the immune response. Also provided is a method of enhancing an immune response in a patient, comprising administering to the patient a therapeutically effective amount of the ...

[0265] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 is an anti-P It should be administered to a patient in need thereof in combination with the D-L1 antibody for a period of time sufficient to enhance the immune response. Also provided is a method of enhancing an immune response in a patient, comprising administering to the patient a therapeutically effective amount of the ...

[0266] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P It should be administered to a patient in need thereof in combination with the D-L1 antibody for a period of time sufficient to enhance the immune response. Also provided is a method of enhancing an immune response in a patient, comprising administering to the patient a therapeutically effective amount of the ...

[0267] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. In combination with anti-PD-1 antibodies, it will be used for a sufficient period of time to treat colorectal cancer in patients who need it. Also provided is a method of treating a patient with colorectal cancer comprising administering

[0268] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. In combination with anti-PD-L1 antibodies, for a sufficient period of time to treat colorectal cancer patients who need it Also provided is a method of treating a patient having colorectal cancer, comprising administering to

[0269] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. In combination with anti-PD-L2 antibodies, for a sufficient period of time to treat colorectal cancer, patients who need it Also provided is a method of treating a patient having colorectal cancer, comprising administering to

[0270] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. and administering it to patients in need of it for a period of time sufficient to treat lung cancer. Also provided is a method of treating a patient with lung cancer, comprising administering to the patient a therapeutically effective amount of ... pulmonary fibrosis inhibitor.

[0271] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. and administering it to patients in need of it for a period of time sufficient to treat lung cancer. Also provided is a method of treating a patient with lung cancer, comprising:

[0272] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. and administering it to patients in need of it for a period of time sufficient to treat lung cancer. Also provided is a method of treating a patient with lung cancer, comprising:

[0273] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. This drug will be administered in combination with an anti-PD-1 antibody for a period of time sufficient to treat prostate cancer in patients who need it. Also provided is a method of treating a patient with prostate cancer, comprising administering to the patient a therapeutically effective amount of

[0274] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. In combination with anti-PD-L1 antibodies, it will be used for a sufficient period of time to treat prostate cancer in patients who need it. Also provided is a method of treating a patient with prostate cancer comprising administering

[0275] The present invention also provides a therapeutically effective amount of an antibody that specifically binds CD38 as an antagonist. In combination with anti-PD-L2 antibodies, it will be used for a sufficient period of time to treat prostate cancer in patients who need it. Also provided is a method of treating a patient with prostate cancer comprising administering

[0276] Anti-LAG-3 antibodies that enhance the immune response may be used in the methods of the invention. Representative anti-LAG-3 antibodies are described, for example, in WO 2010 / 019570. It is something.

[0277] Anti-CTLA-4 antibodies that enhance the immune response may be used in the methods of the invention. A representative anti-CTLA-4 antibody is ipilimumab.

[0278] Anti-PD-1, anti-PD-L1, anti-PD-L2, anti-LAG3, anti- The TIM3 and anti-CTLA-4 antibodies were generated de novo using the methods described herein. This is also fine.

[0279] In some embodiments, an anti-PD1 antibody comprises a VH of SEQ ID NO: 32 and a VL of SEQ ID NO: 33. Antibodies may be used.

[0280] In some embodiments, an anti-PD1 antibody comprises a VH of SEQ ID NO: 34 and a VL of SEQ ID NO: 35. Antibodies may be used.

[0281] In some embodiments, an anti-TIM antibody comprises a VH of SEQ ID NO:36 and a VL of SEQ ID NO:37. -3 antibody may be used.

[0282] In some embodiments, an anti-TIM antibody comprises a VH of SEQ ID NO:38 and a VL of SEQ ID NO:39. -3 antibody may be used.

[0283] SEQ ID NO:32 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQ APGQGLEWMGGIIPIFDTANYAQKFQGRVTITADESTSTA YMELSSLRSEDTAVYYCARPGLAAAYDTGSLDYWGQGTLV TVSS

[0284] SEQ ID NO:33 EIVLTQSPATLSLSPGERATLSCRASQSVRSYLAWYQQK PGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLE PEDFAVYYCQQRNYWPLTFGQGTKVEIK

[0285] SEQ ID NO:34 EVQLVESGGGLVQPGGSLRLSCAASGFAFSRYDMSWVRQ APGKGLESVAYISGGGANTYYLDNVKGRFTISRDNAKNSL YLQMNSLRAEDTAVYYCASPYLSYFDVWGQGTLVTVSS

[0286] SEQ ID NO:35 EIVMTQSPATLSVSPGERATLSCRASQSLSDYLHWYQQK PGQAPRLLIKSASQSISGIPARFSGSGSGTEFTLTISSLQ SEDFAVYYCQNGHSFPYTFGQGTKLEIK

[0287] SEQ ID NO:36 EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQ APGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTL YLQMNSLRAEDTAVYYCAKSPYAPLDYWGQGTLVTVSS

[0288] SEQ ID NO:37 EIVLTQSPATLSLSPGERATLSCRASQSVNDYLAWYQQK PGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLE PEDFAVYYCQQGGHAPITFGQGTKVEIK

[0289] SEQ ID NO:38 EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWMQWVRQ MPGKGLEWMGAIYPGDGDIRYTQNFKGQVTISADKSISTA YLQWSSLKASDTAMYYCARWEKSTTVVQRNYFDYWGQGTT VTVSS

[0290] SEQ ID NO:39 DIQMTQSPSSLSASVGDRVTITCKASENVGTFVSWYQQK PGKAPKLLIYGASNRYTGVPSRFSGSGSGTFTLTISSLQ PEDFATYYCGQSYSYPTFGQGTKLEIK

[0291] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P Administered in combination with D-1 antibody to patients in need thereof for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising:

[0292] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-P Administered in combination with D-1 antibody to patients in need thereof for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising:

[0293] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 was an anti-T In combination with the IM-31 antibody, administered to patients in need for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to the patient a therapeutically effective amount of a therapeutically effective amount of a compound selected from the group consisting of ribozyme, ribozyme, cyclodextrin ...

[0294] The present invention also provides a therapeutically effective amount of a CD40 comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. The antibody that specifically binds 38 is an anti-T antibody comprising the VH of SEQ ID NO: 38 and the VL of SEQ ID NO: 39. In combination with the IM-31 antibody, administered to patients in need for a period of time sufficient to treat solid tumors. Also provided is a method of treating a patient having a solid tumor, comprising administering to the patient a therapeutically effective amount of a therapeutically effective amount of a compound selected from the group consisting of ribozyme, ribozyme, cyclodextrin ...

[0295] In the methods of the invention, the combination of an antibody that specifically binds CD38 and a second therapeutic agent is optionally For example, antibodies that specifically bind CD38 and The two therapeutic agents may be administered to a patient on the same day and in the same intravenous infusion, except that CD38 The antibody that specifically binds and the second therapeutic agent may also be administered every other day or every other week or every other month, etc. In some embodiments, an antibody that specifically binds CD38 and a second therapeutic agent that they are simultaneously present (e.g., in serum) at detectable concentrations in the treated patient. In some methods, the administration may be performed in a number of doses over a period of time. The entire course of treatment with an antibody that specifically binds CD38, consisting of multiple doses, This is followed or preceded by a course of treatment with a second therapeutic agent consisting of three or more A recovery period of 1, 2, or several days or weeks may be required to allow the cells to bind CD38 specifically. The administration of the antibody may be used between the administration of the second therapeutic agent and the administration of the antibody.

[0296] An antibody that specifically binds CD38 or an antibody that specifically binds CD38 and a second therapeutic agent Combination with any form of radiation therapy (external beam radiation therapy, intensity-modulated radiation therapy (IMRT) , focused radiation therapy, etc.), and any form of radiosurgery (Gamma Knife, Cyberknife, Linac, and interstitial radiation (e.g., radioactive seed implants, Glia Si te balloon), and / or in conjunction with surgery.

[0297] The focused radiation techniques that may be used include stereotactic radiosurgery, fractionated stereotactic radiosurgery, and focused radiosurgery. Modulated radiation therapy (IMRT) and stereotactic radiosurgery are used to treat tumors such as brain tumors. In this case, radiation is delivered precisely to the tumor-bearing tissue while avoiding the surrounding tumor-free normal tissue. It is clear that the dose of radiation delivered by stereotactic radiosurgery varies. Typically, the dose is from 1 Gy to about 30 Gy, for example, from 1 to 5, 10, 15, 20, 2 5, and may include intermediate ranges in dose, including up to 30 Gy. Thanks to the MRI scan, stereotactic radiation does not have to be delivered in one treatment. Treatment planning can be done day by day. Can be reliably replicated, thereby allowing for the delivery of multiple fractionated doses of radiation When radiosurgery is used to treat tumors over time, such radiation The treatment method is called "fractionated stereotactic radiosurgery," or FSR. In contrast, stereotactic radiosurgery refers to treatment in a single session. Fractionated stereotactic radiosurgery has a high cure rate, i.e. Resulting in a high rate of tumor cell killing and low impact on normal tissue Tumors and normal tissues are more sensitive to radiation, with one high dose being delivered versus multiple low doses. Each responds differently to multiple exposures. , potentially killing more normal tissue than multiple, lower doses of radiation. Therefore, multiple low doses of radiation can be used to treat more cancer while sparing healthy tissue. The dose of radiation delivered by fractionated stereotactic radiosurgery can kill many tumor cells. The amount can vary from 1 Gy to about 50 Gy, for example, 1 to 5, 10, 15, 20, 25 , 30, 40, and intermediate ranges in hypofractionated doses including up to 50 Gy. Intensity-modulated radiation therapy (IMRT) can also be used. IMRT uses high-precision, three-dimensional conformal irradiation. This is an advanced mode of 3D-CRT using a computer-controlled linear accelerator. , which delivers precise doses of radiation to a malignant tumor or to specific areas within a tumor. In a CRT, the profile of each radiation beam is determined by a multi-leaf collimator. (MLC) to fit the target profile from the beam direction image (BEV). The IMRT system reduces the intensity of the radiation beam by 30%. By modulating the radiation dose into multiple small doses, the radiation dose can be more precisely targeted to the three-dimensional (3D) shape of the tumor. Therefore, IMRT allows for close coverage of the ionizing radiation to nearby normal and vital structures. This allows higher radiation doses to be focused on areas within the tumor while minimizing the radiation dose to the tumor. IMRT is used, for example, when the tumor is near vulnerable structures such as the spinal cord, major organs, or blood vessels. When wrapped around the body, it improves the ability to adapt the treatment volume to the concave tumor shape.

[0298] Subcutaneous administration of a pharmaceutical composition comprising an antibody that specifically binds CD38 and hyaluronidase The antibody that specifically binds CD38 is an antibody that specifically binds CD38 and a hyaluronan antibody. The compound may be administered subcutaneously as a pharmaceutical composition comprising the agonist, agonist, and an agonist.

[0299] The concentration of the antibody that specifically binds CD38 in the pharmaceutical composition to be subcutaneously administered is about 20 mg / mL. / mL.

[0300] The pharmaceutical composition to be administered subcutaneously contains approximately 1,200 mg to 1,800 mg of CD38 specific The antibody may comprise an antibody that binds to

[0301] The pharmaceutical composition for subcutaneous administration contains approximately 1,200 mg of an antibody that specifically binds CD38. It may include.

[0302] The pharmaceutical composition for subcutaneous administration contains approximately 1,600 mg of an antibody that specifically binds CD38. It may include.

[0303] The pharmaceutical composition for subcutaneous administration contains approximately 1,800 mg of an antibody that specifically binds CD38. It may include.

[0304] The pharmaceutical composition to be administered subcutaneously contains about 30,000 U to 45,000 U of hyaluronidase. It may contain ze.

[0305] The pharmaceutical composition to be administered subcutaneously contains approximately 1,200 mg of an antibody that specifically binds CD38. , and about 30,000 U of hyaluronidase.

[0306] The pharmaceutical composition to be administered subcutaneously contains approximately 1,800 mg of an antibody that specifically binds CD38. , and about 45,000 U of hyaluronidase.

[0307] The pharmaceutical composition to be administered subcutaneously contains approximately 1,600 mg of an antibody that specifically binds CD38. , and about 30,000 U of hyaluronidase.

[0308] The pharmaceutical composition to be administered subcutaneously contains approximately 1,600 mg of an antibody that specifically binds CD38. , and about 45,000 U of hyaluronidase.

[0309] The pharmaceutical composition to be administered subcutaneously comprises a hyaluronidase having the amino acid sequence of SEQ ID NO:40. It may contain zerHuPH20.

[0310] rHuPH20 is a recombinant hyaluronidase (HYLENEX® recombinant). and is described in International Patent Publication No. WO 2004 / 078140.

[0311] Hyaluronidase breaks down hyaluronic acid (EC 3.2.1.35) and mediates the synthesis of extracellular matrix proteins. This reduces the viscosity of the hyaluronan in the tissue, thereby increasing tissue permeability.

[0312] SEQ ID NO:40 MGVLKFKHIFFRSFVKSSGVSQIVFTFLLIPCCLTLNFR APPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSP RINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKIS LQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWK PKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDF LVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCF NVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVR NRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQD ELVYTFGETVALGASGIVIWGTLSIMRSMKSCLLLDNYME TILNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLH LNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYS TLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQ IFYNASPSTLSATMFIVSILFLIISSVASL

[0313] The administration of a pharmaceutical composition comprising an antibody that specifically binds CD38 and hyaluronidase is Days, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks It may be repeated after 7 weeks, 2 months, 3 months, 4 months, 5 months, 6 months or more. Repeated courses of treatment are possible, as are long-term administrations. The doses may be the same or different. For example, A pharmaceutical composition containing an antibody binding to the antibody and hyaluronidase was administered once a week for 8 weeks, followed by administration twice a week for 2 weeks. The pharmaceutical composition may be administered once every 4 weeks for 16 weeks. 00 mg of an antibody that specifically binds CD38, and approximately 30,000 U of hyaluronidase. and the concentration of the antibody that specifically binds CD38 in the pharmaceutical composition may be about 20 mg / mL or less. The pharmaceutical composition to be administered contains approximately 1,800 mg of a compound that specifically binds CD38. and about 45,000 U of hyaluronidase. The product contains approximately 1,600 mg of an antibody that specifically binds CD38 and approximately 30,000 U of hemagglutinin. The pharmaceutical composition administered may comprise about 1,600 mg of CD38. and about 45,000 U of hyaluronidase.

[0314] A pharmaceutical composition comprising an antibody that specifically binds CD38 and hyaluronidase is administered into the abdomen. It may be administered subcutaneously.

[0315] A pharmaceutical composition comprising an antibody that specifically binds CD38 and hyaluronidase in a total amount of about It may be administered in 80 mL, 90 mL, 100 mL, 110 mL, or 120 mL.

[0316] For administration, 25 mM sodium acetate, 60 mM chloride were added prior to administration of the mixture to the subject. Sodium, 140 mM D-mannitol, 0.04% polysorbate 20, pH 5. A 20 mg / mL antibody that specifically binds CD38 was added to 10 mM L-histidine , 130 mM NaCl, 10 mM L-methionine, 0.02% polysorbate 80, Mix with 1.0 mg / mL (75-150 kU / mL) rHuPH20 at pH 6.5. That's fine.

[0317] Although the invention has been described in general terms, embodiments of the invention are set forth in the appended claims. This is further disclosed in the following examples, which should not be construed as limiting.

[0318] Further embodiments of the present invention The following are further embodiments of the present invention, in accordance with the disclosure elsewhere herein. The features of the embodiments of the invention described above as relating to the invention disclosed in the specification are It also relates to each and every one of these further numbered embodiments. 1. An antibody that specifically binds CD38 for use in the treatment of patients with solid tumors. 2. CD38 for use in treating patients with Treg-mediated diseases An antibody that specifically binds. 3. Administering a therapeutically effective amount of an antibody that specifically binds CD38 to a subject. for use in treating patients with myeloid-derived suppressor cell (MDSC)-mediated diseases An antibody that specifically binds CD38. 4. Contacting regulatory T cells (Tregs) with an antibody that specifically binds CD38 An antibody which specifically binds CD38 for use in suppressing the activity of regulatory T cells, comprising: 5. Myeloid-derived suppressor cells (MDSCs) were treated with an antibody that specifically binds CD38. specifically binds CD38 for use in inhibiting the activity of MDSCs, including contacting the antibody. 6. By administering to a patient an antibody that specifically binds CD38, the control CD40 for use in treating patients with solid tumors, including reducing the number of regulatory T cells. An antibody that specifically binds 38. 7. By administering to a patient an antibody that specifically binds CD38, bone marrow tumors in the patient are reduced. The present invention relates to a method for treating patients with solid tumors, the method comprising: An antibody that specifically binds CD38 for use in the treatment of patients. 8. Use according to any of embodiments 1 to 7, wherein the antibody induces an immune response in the patient. An antibody that specifically binds CD38 to 9. Any of embodiments 1 to 8, wherein the immune response is an effector T cell (Teff) response. or an antibody that specifically binds CD38 for use according to. 10. Teff response is CD4 + T cells or CD8 + T cell-mediated embodiment 10. An antibody that specifically binds CD38 for use according to any one of aspects 1 to 9. 11. Teff response is CD8 + According to embodiment 9 or 10, the T cell-mediated An antibody that specifically binds CD38 for use in 12. Teff response is CD8 + Increased T cell count, CD8 + Increased T cell proliferation, T cells Increased clonal expansion, CD8+ Increased memory cell formation, increased antigen-dependent antibody production, or is an increase in cytokine, chemokine or interleukin production. 13. An antibody that specifically binds CD38 for use according to any of the methods set forth in claim 11 to 12. 13. Any of embodiments 1 to 13, wherein the antibody inhibits the function of immune suppressor cells. For use therefore an antibody which specifically binds CD38. 14. Immune suppressor cells are regulatory T cells (Treg) or myeloid-derived suppressors The CD38 cell (MDSC) for use according to any one of embodiments 1 to 14. An antibody that specifically binds. 15. Tregs are CD3 + CD4 + CD25 + CD127 dim T cells, 16. An antibody that specifically binds CD38 for use according to claim 15. 16. For use according to embodiment 14 or 15, in which Tregs express CD38 An antibody that specifically binds CD38. 17. The method according to any one of embodiments 13 to 16, wherein the function of Treg is inhibited by killing Treg. An antibody that specifically binds CD38 for use according to any of the following: 18. Treg killing is induced by antibodies specifically binding to CD38. body-dependent cell cytotoxicity (ADCC), antibody-dependent cell phagocytosis (ADCP), cell-dependent cell For use according to embodiment 17, the An antibody that specifically binds CD38 for the treatment of inflammatory bowel disease. 19. According to embodiment 17 or 18, the killing of Tregs is mediated by ADCC. An antibody that specifically binds CD38 for use. 20. Approximately 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58% or 60% of Tregs killed The method according to claims 17 to 19, 21. MDSCs are CD11b + HLA-DR - CD14 - CD33 + CD15 + cell 15. An antibody that specifically binds CD38 for use according to embodiment 14, which is 22.CD11b + HLA-DR - CD14 - CD33 + CD15 + Cells expressing CD38 22. An antibody that specifically binds CD38 for use according to embodiment 21, wherein the antibody specifically binds CD38. 23. The method of embodiment 21 or 22, in which the function of MDSC is inhibited by killing MDSC. For use therefore an antibody which specifically binds CD38. 24. MDSC killing is induced by antibodies specifically binding to CD38. body-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement-dependent cells For use according to embodiment 23, the An antibody that specifically binds CD38 for the treatment of inflammatory bowel disease. 25. The method according to embodiment 23 or 24, in which the killing of MDSCs is mediated by ADCC. An antibody that specifically binds CD38 for use. 26. Approximately 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, Killing of 52%, 53%, 54%, 55%, 56%, 57%, 58% or 60% of MDSCs 26. A method for the preparation of a medicament for use according to any one of embodiments 23 to 25, comprising: An antibody that 27. The method of embodiment 1, in which the Teff or immune suppressor cells are present in the bone marrow or peripheral blood An antibody that specifically binds CD38 for use according to any of claims 4 to 26. 28. Solid tumors include melanoma, lung cancer, squamous non-small cell lung cancer (NSCLC), non-squamous NSCLC, colorectal cancer, prostate cancer, castration-resistant prostate cancer, stomach cancer, egg Focal cancer, gastric cancer, liver cancer, pancreatic cancer, thyroid cancer, squamous cell carcinoma of the head and neck, esophagus or It is a cancer of the gastrointestinal tract, breast cancer, fallopian tube cancer, brain cancer, urethral cancer, genitourinary cancer, endometriosis, cervical cancer, or The CD38 for use according to any one of embodiments 14 to 27 is a metastatic lesion of cancer. Specific binding antibody. 29. The method according to any one of embodiments 1 to 28, wherein the solid tumor lacks CD38 expression. An antibody that specifically binds CD38 for use in immunohistochemistry. 30. The antibody comprises a heavy chain variable region (VH) of SEQ ID NO: 4 and a light chain variable region (VH) of SEQ ID NO: 5. L) for binding to CD38. For use therefore an antibody which specifically binds CD38. 31. The antibody comprises at least the region SKRNIQFSCK of human CD38 (SEQ ID NO: 1). NIYR (SEQ ID NO: 2) and the region EKVQTLEAWVIHGG (SEQ ID NO: 3) An antibody that specifically binds CD38 for use according to any one of embodiments 1 to 30. body. 32. The antibody comprises heavy chain complementarity determining region (HCDR) 1 of SEQ ID NO: 6, 7, and 8, respectively. (HCDR1), 2 (HCDR2) and 3 (HCDR3) sequences, 5. An antibody that specifically binds CD38 for use according to any of the preceding claims. 33. The antibody comprises a light chain complementarity determining region (LCDR) of SEQ ID NO: 9, 10, or 11, respectively. Embodiments 1 to 3, comprising 1 (LCDR1), 2 (LCDR2) and 3 (LCDR3) sequences 2. An antibody that specifically binds CD38 for use according to either of the methods. 34. The antibody comprises HCDR1, HCD1 and HCD2 of SEQ ID NOs: 6, 7, 8, 9, 10 and 11, respectively. R2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences. 33. An antibody that specifically binds CD38 for use according to any of the preceding claims. 35. The antibody is 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:4. and a VH that is 95%, 96%, 97%, 98%, 99% or 100% identical to SEQ ID NO:5. 35. CD38 for use according to any of embodiments 1 to 34, comprising a VL that is 100% identical to An antibody that specifically binds. 36. Any of embodiments 1 to 31, wherein the antibody comprises a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5. An antibody that specifically binds CD38 for use according to any of the following: 37.Antibodies, a. VH of SEQ ID NO: 14 and VL of SEQ ID NO: 15; b. VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c. VH of SEQ ID NO: 18 and VL of SEQ ID NO: 19, or d. HCDR1, HCDR2, HC 30. Any of embodiments 1 to 29, including LCDR1, LCDR2 and LCDR3. For use therefore an antibody which specifically binds CD38. 38.Anti-CD38 antibody, a. VH of SEQ ID NO: 14 and VL of SEQ ID NO: 15; b. VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c. VH of SEQ ID NO: 18 and VL of SEQ ID NO: 19, or d. Use according to embodiment 37, comprising a VH of SEQ ID NO: 20 and a VL of SEQ ID NO: 21 An antibody that specifically binds CD38 to 39. The method according to any one of embodiments 1 to 38, wherein the antibody is administered in combination with a second therapeutic agent. An antibody that specifically binds CD38 for use in 40. The second therapeutic agent is a chemotherapy drug, targeted anti-cancer therapy, 39. A standard of care drug for the treatment of solid tumors, or an immune checkpoint inhibitor. An antibody that specifically binds CD38 for use according to. 41. Immune checkpoint inhibitors include anti-PD-1 antibodies, anti-PD-L1 antibodies, and anti-PD- In embodiments, the antibody is an anti-L2 antibody, an anti-LAG3 antibody, an anti-TIM3 antibody, or an anti-CTLA-4 antibody. 40. An antibody that specifically binds CD38 for use according to 42. According to embodiment 39, the second therapeutic agent is administered simultaneously, sequentially or separately. An antibody that specifically binds CD38 for use. 43. According to any of embodiments 39 to 42, the patient is treated with radiation therapy. An antibody that specifically binds CD38 for use. 44. A VH of SEQ ID NO: 4 and a VH of SEQ ID NO: 5 for use in treating patients with solid tumors. An antibody that specifically binds CD38 comprising the VL of 45. The antibody is administered in combination with an anti-PD-1 antibody comprising a VH of SEQ ID NO: 22 and a VL of SEQ ID NO: 23. and VH of SEQ ID NO: 4 for use in the treatment of patients with solid tumors, An antibody that specifically binds CD38 comprising the VL of sequence number 5. 46. ​​The antibody is in combination with an anti-PD-1 antibody comprising a VH of SEQ ID NO: 24 and a VL of SEQ ID NO: 25. and VH of SEQ ID NO: 4 for use in the treatment of patients with solid tumors, An antibody that specifically binds CD38 comprising the VL of sequence number 5. 47. VH of SEQ ID NO: 4 and VL of SEQ ID NO: 5 for use in enhancing the immune response of a patient An antibody that specifically binds CD38. 48. The antibody is administered in combination with an anti-PD-1 antibody comprising a VH of SEQ ID NO: 22 and a VL of SEQ ID NO: 23. VH of SEQ ID NO: 4 and SEQ ID NO: 5 for use in enhancing the immune response of a patient, An antibody that specifically binds CD38 containing VL no. 5. 49. The antibody is in combination with an anti-PD-1 antibody comprising a VH of SEQ ID NO: 24 and a VL of SEQ ID NO: 25. VH of SEQ ID NO: 4 and SEQ ID NO: 5 for use in enhancing the immune response of a patient, An antibody that specifically binds CD38 containing VL no. 5. 50. The antibody comprises an anti-PD-L1 antibody comprising a VH of SEQ ID NO: 26 and a VL of SEQ ID NO: 27. VH and VLK of SEQ ID NO: 4 for use in the treatment of patients with solid tumors, and a VL of sequence number 5. 51. The antibody comprises an anti-PD-L1 antibody comprising a VH of SEQ ID NO: 28 and a VL of SEQ ID NO: 29. VH and VLK of SEQ ID NO: 4 for use in the treatment of patients with solid tumors, and a VL of sequence number 5. 52. The antibody comprises an anti-PD-L1 antibody comprising a VH of SEQ ID NO: 30 and a VL of SEQ ID NO: 31. VH and VLK of SEQ ID NO: 4 for use in the treatment of patients with solid tumors, and a VL of sequence number 5. 53. The antibody comprises an anti-PD-L1 antibody comprising a VH of SEQ ID NO: 26 and a VL of SEQ ID NO: 27. VH and sequence of SEQ ID NO: 4 for use in enhancing the immune response in a patient, administered in combination An antibody that specifically binds CD38 containing the VL of number 5. 54. The antibody comprises an anti-PD-L1 antibody comprising a VH of SEQ ID NO: 28 and a VL of SEQ ID NO: 29. VH and sequence of SEQ ID NO: 4 for use in enhancing the immune response in a patient, administered in combination An antibody that specifically binds CD38 containing the VL of number 5. 55. The antibody comprises an anti-PD-L1 antibody comprising a VH of SEQ ID NO: 30 and a VL of SEQ ID NO: 31. VH and sequence of SEQ ID NO: 4 for use in enhancing the immune response in a patient, administered in combination An antibody that specifically binds CD38 containing the VL of number 5. EXAMPLES

[0319] Example 1. General Materials and Methods Sample collection and processing Peripheral blood and bone marrow aspirates were collected at baseline immediately prior to the first infusion and at specific time points during treatment. The samples were collected in heparinized tubes. Upon arrival at the central laboratory 24–48 hours after collection, the samples were The majority of peripheral blood mononuclear cells (PBMs) were assessed using real-time flow cytometry. C) were obtained from whole blood, isolated by density gradient centrifugation, and stored frozen until analysis. Cell activation, clonality, and CD38 + For the Treg suppression assay, frozen PBMC samples were Using the pool, pre- and post-treatment samples were analyzed simultaneously.

[0320] NK, T, B, myeloma cells (CD138 + ) and CD38 expression were evaluated in advance. BARC global central la Samples were analyzed by flow cytometry at the laboratory. The fluid and bone marrow samples were subjected to the following multi-fluorochrome antibody panels: erCPCy5.5α-CD19(cloneHIB19;Becton Dickin son[BD]), APCα-CD24(SN3;eBioscience), PC7α -CD3(UCHT-1;Beckman Coulter), V500α-CD16( 3G8; BD), and PEα-CD56 (MY; BD); regulatory T cells (T reg ) Panel Le: APCα-CD25 (2A3; BD), PEα-CD127 (HIL-7R-M21 ;BD), APC-H7α-HLA-DR(G46-6;BD), and PerCPα-C D4 (L200; BD); Naive / Memory T cell panel: APC-H7α-CD4 ( RPA-T4;BD), PerCP-Cy5.5α-CD8(RPA-T4BD), PE α-CD62L (SK11; BD), and APCα-CD45RA (HI100; BD) CD38 expression was stained with the VH and VL sequences of SEQ ID NO: 14 and SEQ ID NO: 15. Alexa 647-labeled antibody mAb 00 described in U.S. Pat. No. 7,829,693 3. Blood samples were prepared using various Lyse-wash methods. Bone marrow aspirate samples were subjected to either membrane or intracellular staining with a variety of antibodies. The Becton Dickinson FACS Microsystem was used to lyse red blood cells in peripheral blood samples. Lysing solution was used for intracellular staining of bone marrow aspirate samples. Fix and Perm cell permeabilization reagent was used Stained samples were obtained on a FACS Canto II flow cytometer and analyzed by FACS Data were analyzed using sDiva software. The log count and percentage of lymphocytes in bone marrow samples were determined at all time points examined.

[0321] T cell receptor (TCR) sequencing T cell diversity was analyzed by deep sequencing of TCR rearrangements and PBM Genomic DNA from C samples was used to determine CD8 + T cell clonality was assessed. TCR sequencing is commercially available from Adaptive Biotechnologies. The analysis was performed using the Immunoseq™ assay, which is a pre-qualified multiplex assay. Rex Polymerase Chain Reaction (PCR) Assay (TR2015CRO-V-019) (Variable (V) genes (forward primer) and joining (J) genes (reverse primer) (consisting of forward and reverse primers that directly target the family of Each V and J gene primer was used to amplify somatically recombined TCR. Each primer acts as a priming pair for amplifying a specific universal DNA sequence. After the first round of PCR amplification, each amplicon contained DNA Forward and reverse primers containing universal and adapter sequences required for sequencing. A second round of amplification was performed using amplicons.

[0322] T cell responses to viral and alloantigens Patient PBMCs were seeded in 96-well plates (2 × 10 5 cells / well), human cells Viruses such as cytomegalovirus (CMV), Epstein-Barr virus (EBV), and influenza Restricted by 23 major histocompatibility complex (MHC) class I antigens derived from the virus Viral peptide cocktail (2 μg / mL; CEF peptide pool; PANATecs ( (registered trademark)) or the same number of allogeneic PBMCs from healthy donors irradiated at 25 Gy. The cells were stimulated for 1 day. Unstimulated PBMCs and anti-CD3 / CD28-coated beads were PBMCs stimulated with 100% DMSO were used as negative and positive controls, respectively. Interferon-γ (IFN-γ) in the supernatant was measured by sandwich enzyme-linked immunosorbent assay. (ELISA; Human IFNγ ELISA Ready-SET-Go; eBioscience nce) and was used as a surrogate marker of T cell activation.

[0323] Regulatory T cell (Treg) suppression of effector cell function: Carboxyfluorescein is a novel therapeutic agent for the treatment of pulmonary hypertension. Cuccinimidyl (CFSE) dilution assay PBMCs from healthy donors were cultured using PerCP-Cy5.5α-CD3 (SK7; BD) , KOα-CD45 (J33; Beckman Coulter), V450α-CD4 (SK3; BD), PEα-CD25 (M-A251, BD), PECy7α-CD12 7 (HIL-7R-M21; BD), and APCα-CD38 (HB-7; BD). The effector cells were sorted using a FACS Aria (BD). The cells were then incubated with carboxyfluorescein succinimidyl ester (CFSE; eBioscien e) and CD38 in RPMI + 10% fetal bovine serum. + Treg or CD38 - T Anti-CD3 / C in the presence or absence of Treg (Treg to effector cell ratio 1:1) After 72 hours, cells were stimulated with D28-coated beads. was performed and the dilution percentage of CFSE was used as a surrogate for T cell proliferation.

[0324] Phenotyping of Myeloid-Derived Suppressor Cells (MDSC) and DARZALEX™ Daratumumab-mediated ADCC PBMCs from three normal healthy donors were cultured with myeloma tumor cell lines (RPMI8226, U266, H929) for 6 days, and granulocytic MDSC (G-MDSC) (CD11 b + CD14 - HLA - DR - CD15 + CD33 + ) production was evaluated (Gor Gunn et al., Blood 121:2975-87, 2013). MDSCs were not present in normal healthy PBMCs, but were associated with all three myeloma cell lines. After coculture with PBMCs, G-MDSCs were present as 5–25% of the total PBMC population (data not shown). The gating strategy for flow cytometric evaluation of G-MDSCs was 1 Gate CD11b + followed by CD14 - and HLA -DR - Goethe ing, then CD15 + and CD33 + The gating of GM DSC correlates CD38 expression levels with DARZALEX™ (daratumumab)-mediated AD The cells were sorted and evaluated for their sensitivity to CC. To evaluate the effect of DARZALEX™ (daratumumab) on DCC / CDC Therefore, serum containing complement or an isotype control was included in the ADCC assay.

[0325] Naïve and memory T cell analysis Prior to each infusion of DARZALEX™ (daratumumab), a heparinized peripheral blood sample was Peripheral blood mononuclear cells (PBMCs) were isolated from patients using Ficoll-Hypaque density gradient centrifugation. The cells were isolated by separation and cryopreserved in liquid nitrogen in a medium (10% human serum and 10% dimethyl For FACS analysis, PBMCs were thawed and stored in RPMI supplemented with DMSO. , 2×10 6 Cells / panels were cultured in phosphate-buffered saline containing 0.05% azide and 0.1% HAS. The cells were resuspended in physiological saline (PBS).

[0326] 4. Data Analysis All data analyses and related graphs were performed using R software (R:A Languag e and Environment for Statistical Computing ing,R Development Core Team,R Foundation for Statistical Computing,Vienna,Austri a,2011,ISBN 3-900051-07-0 http_ / / _www_R- All responses that could be evaluated were All treated subjects were included in the data analysis. Consistently, responders were the most Good responses were defined as subjects with sCR, VGPR, and PR, and non-responders were Best response by IRC is defined as subjects with MR, SD, and PD.

[0327] Various statistical comparisons included: (i) baseline levels between responders and non-responders; (ii) baseline versus on-treatment for responders and non-responders; (iii) (iv) percent change during treatment versus baseline; For each comparison, we first performed the Shapiro-Wilk test (Royston (199 5)Remark AS R94:A remark on Algorithm AS 181:The W test for normality.Applied St Normality tests using statistical methods (44, 547-551) have been cited. As a test of the level of difference, we used a non-normal distribution test. Parametric Wilcoxon rank sum test (Hollander and Wolf e(1973), Nonparametric Statistical Method s.New York:John Wiley & Sons.Pages 27~33 (one-sample), 68~75 (two-sample) and Box-Cox variant Transformed t-test (Weisberg, S. (2014) Applied Linear R egression,Fourth Edition,Wiley Wiley,Cha pter 7). The Box-Cox transformation sets A decimal point (1e-07) was added to the new value. In all cases, the two tests were in agreement. The p-values ​​for the Wilcoxon rank sum test are shown in tables throughout the specification unless otherwise stated. Differences between baseline and on-treatment were examined for responders and non-responders. When this is the case, a two-group paired test is performed for the subject, and in all other cases, Independent testing was performed.

[0328] Samples for analysis of various lymphocyte populations were collected simultaneously for different dosing schedules. Population modeling was performed because data were not collected at a single time point. Population modeling for total and activated NK cells was performed using a broken-stick model. We also included fits to the Statistical Model (Lutz et al., "Statistical model el to estimate a threshold dose and its confidence limits for the analysis of su blinear dose-response relationships, exem plified for mutagenicity data.”Mutation Research / Genetic Toxicology and Environm ental Mutagenesis 678.2(2009):118~122). A linear mixed-effects model with intercept and slope was used to estimate the B cell, T cell subpopulation, and leukocyte , monocytes, neutrophils, and lymphocytes were fitted to patient population data (Bates et al. al., (2014). “lme4:Linear mixed-effects mo dels using Eigen and S4.”ArXiv e-print;J Posted in ournal of Statistical Software, http:_ / / _arxiv_org / abs / _1406.5823). Relative days after the start of treatment (A We performed this linear mixed modeling in the log-transformed response variable. We fitted log-scale mixed models. When the response variable value was equal to zero, we fitted the model on the log scale. All response variable values ​​were added with 0.1 to allow for ringing.

[0329] Example 2. Design of Study 54767414MMY2002 (SIRIUS) The target population in study 54767414MMY2002 (SIRIUS) was Patients have received at least three prior lines of therapy, including a PI and an IMiD. or patients with advanced multiple myeloma with dual resistance to a PI and an IMiD. The primary endpoint / final analysis was based on the 2011 IMWG guidelines. Based on Independent Review Committee (IRC) evaluation and computerized algorithms (Clinical Study Report: At least three prior therapies (including proteasome inhibitors and IMiDs) or multiple myeloma with dual resistance to proteasome inhibitors and IMiDs Efficacy and Clinical Trials of DARZALEX™ (Daratumumab) in Subjects with Multiple Myeloma An open-label, multicenter, phase 2 study to investigate efficacy and safety (EDMS-ERI-92399922; Weers et al.,(2011)J Immunol 186(3):184 0~1848).

[0330] These evaluations included overall response rate (ORR), duration of response, time to response and best response, clinical outcome, and Clinical benefit, time to progression (TTP), progression-free survival (PFS), and overall survival (OS ) was included.

[0331] A total of 124 subjects were randomized to receive DARZALEX™ (daratumumab) in this study. Treated (de Weers et al., (2011) J Immunol 186 (3):1840-1848). Eighteen subjects were treated with 8 mg / kg, and 106 subjects The subjects were treated with 16 mg / kg of 100 mg ...

[0332] Group A: DARZALEX™ (daratumumab) 16 mg / kg: 1 and 2 cycles Cycles 3-6, days 1 and 15 (every other week) , and day 1 of the 7th cycle and thereafter (every 4 weeks). Each cycle was 4 weeks long.

[0333] Group B: DARZALEX™ (daratumumab) 8 mg / kg: 1st cycle onwards, Day 1 (every 4 weeks).

[0334] The primary objective of this study is to determine whether patients with PT-IFN-γ-blockade have received at least three prior lines of therapy, including a PI and an IMiD, or Patients with multiple myeloma whose disease is dually refractory to both PIs and IMiDs In subjects, the efficacy and safety of DARZALEX (Darzalex) as measured by ORR (CR+PR) was The purpose of this study was to determine the efficacy of two treatment regimens (Clinical Study Report: Patients had at least three prior lines of therapy (including a proteasome inhibitor and an IMiD) or Patients with multiple myeloma dually refractory to roteasome inhibitors and IMiDs A non-clinical study to investigate the efficacy and safety of DARZALEX™ (daratumumab) in rats Blinded, multicenter, phase 2 study. EDMS-ERI-92399922).

[0335] Secondary objectives of this study are to evaluate the safety and tolerability of DARZALEX™ (daratumumab). Additional efficacy evaluation (e.g., clinical trials) involving evaluation of tolerability, pharmacokinetics, immunogenicity, and pharmacodynamics. Demonstration of clinical benefit, TTP, PFS, and OS, and the efficacy and safety of DARZALEX™ The aim of the study was to discover biomarkers that predict response to daratumumab. Further information related to the study should be provided in the study protocol (clinical study report: at least three treatments) History of chemotherapy (including proteasome inhibitors and IMiDs) or proteasome inhibitors and DARZA in subjects with multiple myeloma who are dually refractory to IMiDs An open-label, multicenter, Phase 2 study investigating the efficacy and safety of LEX™ (daratumumab). This can be confirmed at EDMS-ERI-92399922.

[0336] In stage 1 of part 1, one subject (6%) responded in the 8 mg / kg group and Five subjects (31%) responded in the 16 mg / kg group. It was expanded to Stage 2 and Part 2 of Part 1.

[0337] In the 16 mg / kg group, 31 subjects achieved a response of PR or better based on IRC assessment, The ORR was 29% (95%CI: 21%, 39%). Three subjects (3%) had an sCR. VGPR or better was achieved in 13 subjects (12%).

[0338] Example 3. Patients enrolled in the 54767414MMY2002 study (SIRIUS) DARZALEX™ (daratumumab) on T cell proliferation and activity in patients with Effect CD38 is expressed on a variety of immune and hematopoietic cells. We performed broad immune profiling using DARZALEX against immune cell subsets. (daratumumab) efficacy and clinical response at baseline levels of these cells T cells (CD3 + , CD4 + , CD8 + and regulatory T cells (Treg), B cells (CD19 + ), NK cells, monocytes (CD14 + ), white blood cells, and neutrophils. Cell populations were analyzed in the peripheral blood at baseline and after DARZALEX™ (daratumumab) treatment. Responders and non-responders were assessed by flow cytometry of blood and bone marrow aspirates. The changes in these cell populations in the underculture were monitored.

[0339] Lymphocytes, leukocytes, monocytes and neutrophils Leukocyte, lymphocyte, monocyte, and neutrophil counts were measured in the peripheral blood of responders and nonresponders. In responders, both the 8 mg / kg and 16 mg / kg doses of D ARZALEX™ (daratumumab) treatment was found to increase total lymphocytes. (Figure 1). Linear mixed-effects modeling revealed a 0. 8×10 6 An increase in cells / μL was noted (CI=0.06, 0.11). Monocytes and For white blood cells, the logarithmic scale was 0.03 × 10 6 cells / μ L (CI = 0.01, 0.04) and 0.03 × 10 6 cells / μL (CI=0.01 A slight increase was noted with a significant increase in the median neutrophil count (p < 0.05). The results were consistent with baseline values ​​and did not fluctuate significantly, but neutropenia was observed in some patients. .

[0340] The baseline levels of each of these cell populations were compared between the response groups. Using the signed rank test, no differences in any cell types were observed between response groups at baseline levels. (Table 1).

[0341] [Table 1]

[0342] NK cells Total NK cells (CD16 + CD56 + ) and activated NK cells (CD16 + CD56 dim ) decreased over time with DARZALEX™ (daratumumab) treatment (data not shown). (data not shown).

[0343] B cells B cells (CD45 + CD3 - CD19 + ) absolute number of responses DARZALEX™ (daratumumab) treatment in responders and non-responders B cells were measured over time during treatment. B cells were slightly increased in whole blood and maintained in bone marrow aspirates. Linear mixed modeling of peripheral blood B cell counts was performed on a logarithmic scale for each 100-day period. The minimum increase is 0.1 × 10 6 An increase in cells / μL was observed [CI=0.0 4, 0.16] during DARZALEX™ (daratumumab) treatment. Responders During daratumumab treatment, the B cell count in bone marrow aspirates (CD4 5+ CD3 - CD19 + The proportion of lymphocytes (cells per corpus callosum) did not change (p = 0.1 and p = 0.01, respectively). Furthermore, the difference in baseline B cell counts between responders and non-responders was No significant difference was observed (p=0.5).

[0344] T cells Although B cells showed only a marginal increase (see above), DARZALEX (commercial An increase in lymphocytes was confirmed following treatment with daratumumab (Figure 1). To investigate the role of T cell populations in both peripheral blood and bone marrow, we investigated the different T cell populations (CD3 + , CD4 + , CD8 + T cells, regulatory T cells).

[0345] CD3 + , CD4 + and CD8 + The T cells were treated with DARZALEX™ (daratumumab ) increased in peripheral blood (both absolute counts / μL and percentage of lymphocytes) after treatment. , peripheral blood CD3 in all patients + T cells (CD45 + CD3 + ) absolute number, base The black line in the figure represents the absolute number in all patients × 10 6 cell Median values ​​are shown per μL. Only visits with 3 or more observations were included in the figure. Peripheral blood CD4 + T cells (CD45 + CD3 + CD4 + ) absolute number The black line in the figure indicates the median value for all patients. 3 Only visits with ≥ 1 observation were included in the figure. Medium CD8 + T cells (CD45 + CD3 + CD8 + ) absolute number of changes from baseline over time The black line in the figure indicates the median value for all patients. Only clinic visits were included in the figure. Linear mixed modeling of absolute peripheral blood counts revealed that the mean total T cell counts were significantly higher than the mean total T cell counts. cells (CD45 + CD3 + ) for each 100-day period, on a logarithmic scale, 6 An increase in cells / μL (CI=0.1, 0.15) was observed (DARZALEX (compound Target (after daratumumab treatment). CD8 + T cells increased in logarithmic scale for each 100 days. 0.16×10 6 A significant increase in cells / μL was confirmed (CI=0.13, 0.19 ). CD4 + The cells were 0.11 × 10 on a logarithmic scale for each 100 days. 6 cells / μ A moderate increase in L was identified (CI = 0.09, 0.13).

[0346] For each T cell subpopulation, responders had higher baseline values ​​than non-responders The maximum percentage change in absolute counts was observed for CD3 + p=3.2993e-05;CD4 + p=3.486e-05;CD8 + p=2.7172e-05; regulatory T cells p=0.0 02) Table 2 shows the percentage change in absolute numbers from baseline for responders and non-responders. Results of Wilcoxon signed rank test for comparison of peripheral blood T cell subpopulations between subjects show.

[0347] [Table 2]

[0348] Similarly, in the bone marrow, total T cells (CD45 + CD3 + , as a percentage of lymphocytes) and CD 8 + T cells (CD45 + CD3 + CD8 + , as a percentage of lymphocytes) were In both patients and non-responders during DARZALEX™ (daratumumab) treatment It was found that the CD3 + Responder p=3.8147e-06, No Responder p=9.8225e-05; CD8 + Responder p=3.8147e- 06, non-responder p=0.0003). In both bone marrow clinical response groups, CD 4 + The median T cell count did not change. Table 3 shows the various T cell counts as % lymphocytes in bone marrow. The results of the Wilcoxon signed rank test for DARZALEX (commercial CD45 over time during daratumumab treatment + CD3 + The percentage of cells is shown (%). (Both responders and non-responders are included in the graph.) Figure 6 shows the CD45 over time during (TM)(daratumumab) treatment + CD3 + CD8 + Percentage of cells shown (Both responders and non-responders are included in the graph).

[0349] [Table 3]

[0350] Both responder and non-responder groups showed increased T cells in peripheral blood and bone marrow However, responders had the greatest percentage change from baseline. In responders, CD3 + , CD4 + and CD8 + Identify differences in T cell levels To assess the efficacy and safety of daratumumab in patients with glaucoma, peripheral blood samples were collected from each subgroup prior to DARZALEX™ (daratumumab) treatment. Baseline measurements were compared.

[0351] Absolute T cell counts in peripheral blood (Table 4) or percentage of T cells among total lymphocytes in bone marrow There was no statistically significant difference between responders and non-responders in Table 5. (Wilcoxon signed-rank test).

[0352] [Table 4]

[0353] [Table 5]

[0354] T regulatory cells Treg cells are CD3 + CD4 + CD25 + CD127 dim Cell populations It was identified as CD8 + The ratio of T cells to Tregs was measured using DARZALEX (trademark ) (daratumumab) over time in peripheral blood and bone marrow of patients treated with daratumumab. The increase was seen both in the periphery and bone marrow. CD8 + / Treg and CD8+ / CD4 + Figure 7B shows the median cell ratio in bone marrow. CD8 of all patients at that time + / Treg and CD8 + / CD4 + Median T cell ratio CD8 + Tregs and CD8 + / CD4 + The absolute ratio of The results were significant in peripheral blood (Table 6) and bone marrow (Table 7) (Wilcoxon signed rank test). ).

[0355] A comprehensive data analysis of the SIRIUS and GEN501 studies (Example 6) showed that peripheral blood CD8 + / CD4 + and CD8 + The median ratio of Treg cells was 1.0 at week 8 (p=5.1 ×10 -5 (CD8 + / CD4 + ) and p = 1.8 × 10 -7 (CD8 + / Treg), And at week 16 (p=0.00017(CD8 + / CD4 + ) and p = 4.1 × 10 - 7 (CD8 + Similarly, in the bone marrow, CD8 + / CD4 + and CD8 + The median proportion of Treg cells was 0.01% during treatment (week 12 ± 1 week) compared to baseline. increased (p=0.00016(CD8 + / CD4 + and p = 2.8 × 10 - 7 (CD8 + No significant difference was observed between responders and non-responders. There wasn't.

[0356] [Table 6]

[0357] [Table 7]

[0358] Example 4. Study Design (GEN501) Study GEN501 (NCT00572488) is investigating the efficacy and safety of cyclophosphamide in patients with dual-refractory MM. DARZALEX™ (daratumumab) was evaluated as monotherapy. Sample isolation, The procedures and statistical analysis were as described in Examples 1 and 2. khorst et al.,N Eng J Med 373:1207~19,20 It is stated in 05.

[0359] Briefly, Study GEN501 is evaluating the efficacy and safety of DARZALEX ( This was the first-in-human clinical trial of daratumumab (Daratumumab) in two This is a Phase 1 / 2 dose-escalation safety study divided into two parts. Part 1 is an open-label dose-escalation study. Part 2 is a multi-cohort study based on the dose levels established in Part 1. This is an open-label, single-arm study.

[0360] In part 1, 0.005, 0.05, 0.10, 0.50, 1, 2, 4, 8, 16, Evaluation of DARZALEX™ (daratumumab) at 10 dose levels of 24 mg / kg and The lowest two dose cohorts were assigned one (+3) subject each, and the standard dose cohorts were assigned one (+3) subject each. The allocation of 3 (+3) subjects was applied to the remaining 8 dose cohorts. Part 2 This was an open-label, single-arm study involving two dose levels, 8 mg / kg and 16 mg / kg. Part 1 included 32 subjects and part 2 included 72 subjects.

[0361] Example 5. DARZALEX™ (daratumumab) treatment enhances T cell proliferation in patients Inducing loan-like In the MY2002 study, CD8 + T cell proliferation was confirmed With this in mind, we used the Immunoseq™ assay to identify T cell receptor (TCR) High-throughput next-generation sequencing was performed to identify CD8 + Increased T cells indicate an adaptive immune response The genotypes of subjects enrolled in the GEN501 study were determined to be clonality or non-clonality. A total of 17 patient samples were evaluated (n = 6 were responders, i.e., ≥ PR; n=11 were non-responders, i.e., MR, SD, PD).

[0362] TCR sequencing revealed that DARZALEX™ (daratumumab) treatment improved Figure 8A shows that DA significantly increased clonality across all subjects. Correlation between T cell clonality before versus after RZALEX™ (daratumumab) treatment (p=0.0056). Figure 8B shows the fold change in clonality in individual patients. Responders are marked with an asterisk. This data was obtained using DARZALEX™ (Daratum These results suggest that the T cell expansion observed with mAb treatment may be clonal in nature. There are.

[0363] Compared to non-responders, responders had a significantly altered TCR repertoire (abundance change). The total amount of proliferation (measured as CIA) was greater in each patient. Figure 8C shows the CIA% of each patient. Group A: responders, Group B: non-responders. There was a statistically significant difference between the two (p=0.037). Figure 8D shows the results of each expanded T cell. Absolute change in abundance (CIA) for clones in responders and non-responders The sum of the values ​​is shown. Figure 8E shows the maximum CIA% for individual patients. Group A: Responders, B: Group: Non-responders. No statistically significant difference was observed between responders and non-responders. (p=0.048). Figure 8F shows the relationship between responders (group A) and non-responders (group B). The maximum CIA of a single T cell clone in

[0364] Fisher's exact test (DeWitt et al. J. Virol. 2015 ) and summing the absolute abundance changes for each expanded clone to obtain two CIA was obtained by identifying significant differences in clonal abundance between samples.

[0365] Example 6. DARZALEX™ in Patients Enrolled in the GEN501 Trial Immunomodulatory Effects of Daratumumab In the responders and non-responders enrolled in GEN501, various T and B cell populations were assessed.

[0366] Lymphocytes As in the SIRIUS (MMY2002) study, lymphocytes were treated with DARZALEX ( During treatment with daratumumab, CD4+ increased in both peripheral blood and bone marrow. 4 + and CD8 + This was due to an increase in both cell numbers.

[0367] CD8 + Central Memory Cells In a subset of 17 patients enrolled in the GEN501 trial, CD8 + T cells Phenotypes are examined over time in patients treated with DARZALEX™ (daratumumab) Patient-derived CD8 + Cells were cultured as naïve (CD45RO- / C D62L + )(T N ) or Central Memory (T CM )(CD45RO + / CD62L +high ) cells.

[0368] Figure 9A shows CD8 + Naive cells % (CD8 + Figure 9B shows the percentage of CD8 + S The percentage of central memory cells was 100% with DARZALEX™ (daratumumab) treatment. , naïve CD8 + The amount of T cells was significantly decreased (p=1.82×10 -4 , 8th week) , CD8 + The amount of memory T cells was significantly increased (p=4.88×10 -2 , 8th week This is the process by which naive cytotoxic T cells are transformed into mem- ories that can be activated against specific antigens. White boxes indicate at least minimal response (≥ MR). Black boxes indicate patients with stable disease or progressive disease. + Naive T Thin A significantly greater reduction in cysts was evident in patients who responded to treatment (data not shown). Figure 9C shows that DARZALEX™ (daratumumab) treatment reduced virus-specific The proportion of HLA class I-restricted T cells that partially promote alloreactive T cell responses Figure 9D shows that effector mem- bers expressing low levels of CD38 were increased. These T cells respond to the viral peptides and alloantigens. It is important to note that the α-aminobutyric acid α-aminobutyric acid exhibits normal and even increased functional activity relative to the original (see Example 1). 8). These functional results suggest that There is an increase in or increased activity of antigen-experienced T cells against the virus and alloantigens. These data suggest that effector T cells, as distinct from regulatory cell subsets, These results suggest that cells do not require expression of CD38 to function and proliferate properly. .

[0369] CD38 positive regulatory T cells Recent publications indicate that several immunosuppressive cell subsets express CD38. In addition, the stable proliferation and enhanced activity of cytotoxic T cells were observed, suggesting that the control Regulatory T cells (Treg), myeloid-derived suppressor cells (MDSC), and Effects of DARZALEX™ (daratumumab) on regulatory B cells (Bregs) This prompted consideration of the following:

[0370] Regulatory T cells (Treg) (CD3 + CD4 + CD25 + CD127 dim ) to the standard Tregs were isolated using a quantitative method and the frequency of Tregs was analyzed using flow cytometry.

[0371] A subpopulation of peripheral Tregs (10% ± 10%) expresses high levels of CD3 The top panel of FIG. 10A shows the expression of CD3 + CD4 + cell collection The lower panel of FIG. 10A shows the frequency of Tregs in the P4 cell population. The following Treg subsets express CD38 + Tre g was highly responsive to DARZALEX™ (daratumumab) treatment and After the first dose of ALEX™ (daratumumab), a significant and nearly immediate reduction was observed ( n = 17 patients; P = 8.88 × 10 -16 , at week 1 relative to baseline). The frequency of Tregs after ALEX™ (daratumumab) treatment is shown in the top panel of Figure 10B. The lower panel of FIG. 10B shows CD38 high Treg(P5 thin were most significantly depleted after the first DARZALEX™ (daratumumab) infusion These CD38 + Tregs are The patients remained depleted during treatment with daratumumab (p=8.88×10 -16 , 1. 11×10 -15 , and 1.50×10 -11 , 1, 4, and 8, respectively, relative to the reference value. Figure 10C shows the results of baseline, week 1, week 4, week 8, relapse, and end of treatment (EO CD38 at 6 months after T high Treg% (total CD3 + (from cells) CD38 high Tregs had returned to baseline levels by that time point. + T The changes in reg were similar between responders and non-responders, whereas C D8 +T cell:Treg ratios in response to DARZALEX™ (daratumumab) The mean mean mean ≥ 10 days was significantly higher at week 8 in patients with ≥ 10 days ( P = 0.00955; Fig. 10D ).

[0372] CD38 + Considerations regarding Treg depletion and DARZALEX™ (daratumumab) treatment To assess the biological relevance of CD38 + Tregs vs. CD38 - Tregs, house CD3 + The suppressive ability of T cells was evaluated using samples from multiple healthy donors. In a series of experiments performed, CD38 + Tregs are CD38 - Treg (observed cells (53.2% of cell proliferation) or negative control (74.9% of cell proliferation observed). The cytoplasmic endothelial cells (C-T cells) were cultured in a 10-well platelet-free culture medium, and the ... C-T cells were cultured in a 10-well platelet-free culture medium, and the C-T cells were cultured in a 10-well platelet-free culture medium, and the C-T cells were cultured in

[0373] MDSCs were not readily detectable in frozen PBMC samples, therefore CD38 + Granulocytic MDSC (CD11b + CD14 - HLA-DR - CD15 + CD33 + )of, Patients at baseline and those who received one DARZALEX™ (daratumumab) infusion The identified MDs were produced in vitro from PBMCs isolated from the mice. Flow cytometry histograms of SCs are shown (Figure 11, upper histogram, boxed). Approximately half of the MDSCs expressed CD38 (Fig. 11, center graph; (P7 cell population circled). CD38 high MDSC is a wholly owned subsidiary of DARZALEX™ daratumumab) was almost completely depleted (Figure 11, bottom graph; circled P 7 cell population).

[0374] CD38 high System-nonspecific MDSCs are non-responders and at least In both patients who responded to DARZALEX™ and those who had minimal responders, (Daratumumab) treatment depleted CD38 over time. high MDSC The proportion of patients with refractory pulmonary edema decreased to nearly 0% in the first, fourth, or eighth week of treatment. CD38 high Non-systemic specific MDSCs recovered to baseline levels after treatment.

[0375] CD38 is most abundant in lineage-nonspecific MDSCs + Patients with the population were Patients with daratumumab showed the best and most durable response to daratumumab treatment. , the highest CD38 high MDSC percentage (shown in FIG. 11 ), and PR or M Patients 2, 4, 15, 16, and 17, classified as R patients, were progression-free for at least 8 months. The results show that the patients had a progression-free survival (PFS).

[0376] In addition, CD38 high Lineage-nonspecific MDSCs are The antibodies were also susceptible to in vitro ADCC induced by mAb. ADCC assay CD38 from two donors high MDSC and Dau as control target cells di cells were performed using an effector:target cell ratio of 50:1. The results of the donor experiments are shown. DARZALEX™ (daratumumab) inhibits MDSC cells. This induced the dissolution of

[0377] CD38 + Bregs were compared in patients treated with DARZALEX™ (daratumumab) n=16), CD38 + Like Tregs, DARZALEX( Depletion after the first dose of daratumumab (p=0.0018, 1% vs. baseline) (weekly time point; paired Wilcoxon rank test), remained low while patients were treated. FACS-sorted Bregs were stimulated to secrete IL-10 (Figure 15B).

[0378] Collectively, these observations support the notion that immunosuppressive CD38 + MDSCs, Bregs, and Tres g depletion enhances DARZALEX™ (Daratumma gracilis) effects on T cell populations and clonality These results suggest that this is the mechanism by which oxidative stress significantly induces induced changes.

[0379] Example 7.CD38 + MDSC cells are present in cancer patients MDSC (Lin - CD14 - HLADR low / - ) and their CD38 expression The present study examined peripheral blood samples from patients with NSCLC or prostate cancer using flow cytometry. Defeated.

[0380] The proportion of MDSCs was P The CD38 expression was approximately 10%-37% and approximately 10%-27% of BMC, respectively. Lin derived from human PBMCs - CD14 + HLADR - / low MDSC 80-100 % and 70-100% of MDSCs derived from PBMCs of prostate cancer patients.

[0381] Example 8. DARZALEX™ (daratumumab) enhances antiviral T cell responses To force Effects of DARZALEX™ (daratumumab) on T cell activation and functionality To further evaluate, IFN-γ production from peripheral T cells in response to viral and alloantigens. Patients treated with DARZALEX™ (daratumumab) with mixed clinical outcomes Patients with PR or better were treated with DARZALEX (Daratum After treatment with mAb, viral load and immunization were compared with baseline at at least one time point during treatment. showed a significant increase in IFN-γ secretion in response to alloantigens, and T cell function was improved in mice with low expression of CD38. This suggests that the TCR cloning was not impaired by expression (see Example 6, FIG. 9C). Similar to the efficacy data, this increase was consistent with the response to DARZALEX™ (daratumumab). The difference in the response rate was more pronounced in patients who responded than in those who did not. Figure 16A shows a representative Figure 16B shows the antiviral response of one representative CR patient. Figure 16C shows the antiviral response of one representative PD patient. D shows the antiviral response of one representative MR patient. In the figure, the error bars are doubled. The standard error of the mean of replicate cultures is shown. Asterisks indicate statistically significant differences between the comparisons shown. Means significant change. Best response as determined by an independent review committee. Consistent with these results Virus-reactive T cells in patients with VGPR (Figure 16E) or CR (Figure 16F) They demonstrated increased proliferative capacity during DARZALEX™ (daratumumab) treatment.

[0382] Example 9. DARZALEX™ (Daratumma) of CD38-Expressing Immune Cell Subtypes Mechanism of susceptibility to Data from both the GEN501 and SIRIUS trials support the finding that DARZALEX™ Tumor necrosis factor (TNF) therapy depletes some immune cells that express CD38 (NK cells, regulatory T cells (Treg), regulatory B cells (Breg), and myeloid-derived suppressor cells (MD SC), while the number of other cells expressing CD38 increases (cytotoxic and helper It has been shown that T cells

[0383] To address the susceptibility mechanism, healthy donors and GEN501 or SIRIUS trials were CD38 expression on various immune cell subpopulations in multiple myeloma patients enrolled in the study Figure 17A shows the levels of CD38 expression in immune cells from healthy donors. FIG. 17B shows histograms of CD38 expression in immune cells from multiple myeloma patients. Histograms of CD38 expression are shown. In healthy donors, CD38 expression was highest on NK cells, followed by In patients with multiple myeloma, CD38 expression was highest on plasma cells. The IL-1 expression level was also higher in subsets of B cells, NK cells, monocytes, B cells, and T cells. 7C is a quantitative analysis of NK cells, Tregs, Bregs, B and T cells from patients with relapsed and refractory myeloma. The mean fluorescence intensity (MFI) of CD38 in cells was compared, and NK cells were the first to express CD38 after plasma cells. They also express high levels of CD38, which subsequently mediates the expression of regulatory T cells (Treg) and regulatory B cells ( This indicates that the 1960s was a 1970s Breg.

[0384] In addition to CD38 expression, complement inhibitory proteins (CIPs; CD46, CD55, CD59 ) are involved in the binding of DARZALEX™ (daratumumab) to other cell surface proteins. These findings may contribute to the susceptibility or resistance of some immune cell subpopulations to CIP. By intravitro assay, NK cells express very low levels of CD59 and CD55. However, other T and B cell populations were found to express it at much higher levels. DARZALEX™ (daratumumab) is a multi-target agent that targets multiple immune cell subtypes. may contribute to susceptibility (data not shown).

[0385] Consideration The study is investigating the previously unknown DARZALEX™ (daratumumab ) of CD38 + Reduction of immunosuppressive cell populations and associated responses to viral peptides Induction of helper and cytotoxic T cell proliferation, IFN-γ production, and improved adaptation to Explains the immunomodulatory effects of TCR clonal expansion indicative of immune responses.

[0386] This study demonstrated that MDSCs and Bregs express CD38 and are associated with DARZALEX (commercially available). These cells were sensitive to treatment with daratumumab. They are present in the tumor boundary and contribute to tumor growth, immune evasion, angiogenesis, metastasis, and the production of inhibitory cytokines. These CD38 + In addition to the suppressor cell subset, A novel subpopulation that also expressed high levels of CD38 and exhibited superior suppressive ability against autologous T cells Regulatory T cells (CD4 + CD25 + CD127 dim ) were identified. The cells were also sensitive to DARZALEX™ (daratumumab) and were significantly more prevalent in treated patients. These CD38 + DARZALEX™ immunoregulatory cells Mumab-mediated killing reduces local immune suppression within the myeloma microenvironment and enhances positive immune effects. This could enable proliferation of tumor-specific cytokines (T-cells) and their contribution to the anti-tumor response.

[0387] In fact, CD4 + and CD8 + A significant increase in a broad range of T cell populations was observed, including both CD8 + The subpopulation is D ARZALEX™ (daratumumab) therapy, which includes naive T-cell and a significant decrease in effector memory CD8 + A significant increase in T cells was observed and antigen-experienced phenotype (able to retain immune memory and respond to tumor antigens) This shows the migration of effector T cells to CD8 + :CD4 + and CD8 + :Tre The ratio of g to β-glucose was also significantly increased by treatment, indicating a shift towards a positive for negative immune regulators. is.

[0388] Expanded CD4 + and CD8 + To assess whether T cells were clonal in nature To this end, the T cell repertoire was examined in a subset of patients. DARZALEX™ (daratumumab) is effective in patients with SD or progressive disease. Thus, the increase in T cell clonality was not simply due to a decrease in tumor burden. However, T cell clonal skewing is significant in patients with good clinical responses. , CD8 +The observed DARZALEX™ increase correlated with an increase in T cells This suggests that T cell proliferation due to daratumumab treatment was antigen-driven. This was especially evident in a patient population that had received a large number of prior treatments (median number of prior treatments, 5). In addition to the increase in TCR clonality, it is not expected that the tumor can initiate a strong antitumor immune response. In one study, patients who responded to DARZALEX™ (daratumumab) were randomized to receive 100 mg of pre-existing The results showed that the immune system was immunosuppressed due to increased T cell responses to alloantigens and alloantigens. This suggests that the patient recovered from the condition.

[0389] Treatment with DARZALEX™ (daratumumab) inhibits immunosuppressive MDSCs, as well as These reductions were associated with a decline in the number of CD4 + T-helper Cells and CD8 + This was accompanied by an expansion of cytotoxic T cells, as measured by IFN-γ production. Cellular clonality and functional antiviral responses were also assessed by DARZALEX™ (daratumumab ) treatment. These observations suggest that T cells, despite their low CD38 expression, This indicates that the increased T cell response could be due to depletion of regulatory cells. Furthermore, these changes in T cell proliferation, activity, and clonality are associated with DA Patients who responded compared to those who did not respond to RZALEX™ (daratumumab) Relapse from DARZALEX™ (daratumumab) therapy was more prominent. Treatment with cyclosporine was associated with reversal of many of these changes, which may have implications on clinical response and efficacy. Further, the potential benefits of DARZALEX™ (daratumumab) through immunomodulation may be This suggests a previously uncharacterized mechanism of action.

[0390] Recently, there has been mixed evidence on the effectiveness of antibodies to promote antitumor immune responses rather than directly targeting cancer. Antibodies that block CTLA-4 and PD-1 have been shown to suppress T cell proliferation. By promoting the expression of IL-1 and enhancing T cell activation, it is possible to treat advanced solid tumors and Hodgkin's lymphoma. It improves survival and delays disease recurrence in patients with any hematological malignancy. By boosting immunity, these immunomodulatory antibodies may be able to induce clinical responses. It can also prevent the recurrence of the disease.

[0391] Example 10.54767414MMY2002 (SIRIUS) Part 2 Clinical Trial and traded with DARZALEX™ (daratumumab) as monotherapy. Serum proteomic analysis of multiple myeloma patients Biomarker sample collection and processing Peripheral blood samples were collected in standard serum separator tubes (2.5 mL to 5 mL) and a portion of the serum was removed. were stored frozen in SomaLog for multi-analyte serum protein profiling. The document was sent to ic, Inc. (Boulder, CO).

[0392] Serum protein profiling utilizes affinity-based molecules called SOMAmers The pre-validated SOMAscan assay measures 1129 protein analytes using The SOMAmer reagent was a single-stranded DNA-based This assay is a protein affinity reagent. ) and protein signals are quantified by custom DNA microarrays. Convert to SOMAmer signal.

[0393] Each SOMAmer contains four functional moieties: 1. Unique protein recognition sequence 2. Capture biotin 3. Photocleavable Linkers 4. Fluorescent molecules for detection

[0394] The unique protein recognition sequence uses DNA and chemically modified nucleic acids that mimic amino acid side chains. Incorporating nucleotides to expand the diversity of standard aptamers and to identify protein-nucleic acid interactions This increases the specificity and affinity of action (Gold et al., PLoS One 5 :e15004,2010). Aptamers are selected by SELEX. SOMA The mer reagents are selected using the protein in its native conformation. MAmer reagents require native protein tertiary structure for binding. Undetected or denatured, presumably inactive proteins are not detected by the SOMAmer reagent. stomach.

[0395] SOMAmer reagent master mixes grouped by sample type and dilution Prior to sample incubation, the reagents are preloaded onto the streptavidin beads. Proteins in the sample are allowed to bind to their cognate SOMAmers during equilibration. After washing, incubation with NHS-biotin, washing, the beads were exposed to UV light. The eluate contains the photocleavable linker bound to the biotin-labeled protein. Contains SOMAmer reagent. Streptavidin capture and subsequent washing removes unbound Remove any SOMAmer reagents that may be present in the final eluate. The final eluate is then purified using a custom Agilent DN A microarray and the fluorophores derived from SOMAmer molecules (relative fluorescence units (RFU) ) and hybridize to 100 μl of ... .

[0396] Samples for the MMY2002 study were tested in two main batches. Batch 1 The 180 samples were assigned the corresponding cycle 1 day 1 (C1D1, reference value) and C Serum samples from 90 subjects on 3D1 (day 1 of cycle 3) were included. The samples from the second batch were analyzed together on three separate SomaScan plates. The samples included 50 C1D1 samples, including 35 duplicates from batch 1. Can be enjoyed.

[0397] 4. Data Analysis Input Data Set and Definitions Treated subjects with an evaluable response were included in the data analysis. The best overall response (by IRC in MMY2002) for responders was sCR, VGP Subjects were defined as R, PR, stable disease (SD), minimal response (MR) or SD. The overall best response (based on IRC in MMY2002) was defined as progression (P D) is defined as an object that is

[0398] Preprocessing the Somalogic data Batch Alignment Batches 1 and 2 of the MMY2002 samples were analyzed using two versions of SOMAscan The differences between the two versions are minor, and the differences between the versions are The sequence of the three modified SOMAmers was included (CTSE:3594-6_1 →3594-6_5, FCN1:3613-62_1→3613-62_5, BMPER :3654-27_1→3654-27_4). These were excluded from the analysis.

[0399] The measurements of the three batch 1 plates were performed using standard SomaLogic interplate calibration. Follow-through, seven-place, in-plate control standard measurements of master mixture-specific comprehensive standards The whole plate calibration fold for each SOMAmer was calculated by calculating the ratio of the mean The plate-specific magnification of each SOMAmer reagent was determined by The test was applied equally to each sample on the sheet.

[0400] Considering the different SOMAscan platform versions for batches 1 and 2, By utilizing replicate measurements of 35 samples between tests, SomaLogic A modified standard interplate calibration procedure was performed to correct for systematic batch-to-batch variability. For each SOMAmer, the ratio of the batch 1 post-calibration measurement to the batch 2 pre-calibration measurement was calculated for each of the 35 replicate samples (ri,j). The median value was used to determine the modified SOMAmer-specific calibration fold for the samples in batch 2. Ta

[0401]

number

[0402]

number

[0403] After calculating the modified calibration factor, the distribution of all factors for each analytical batch was plotted and the outliers were The presence or absence of values ​​was evaluated. Nine were with extremely large or small standards (>0.25 and <3). Two SOMAmers were excluded from the analysis due to poor reproducibility.

[0404] After completing batch alignment and SOMAmer selection for MMY2002, Apply a log2 transformation to all protein concentration values ​​in step 2 to make the data more consistent with a normal distribution. and improved the performance of parametric statistical tests.

[0405] Confounding variable adjustment Described by meta-variables (such as demographic factors, response class, and sampling time point) Estimating the fraction of dataset variation that will be affected and identifying possible confounding factors are key steps in centering and This was done by principal component analysis of the scaled and rescaled data set. A simple linear model was applied. We then performed a regression analysis to identify the highest-ranking PCs that were significantly associated with each variable of interest. The significance of these associations was determined using the Wald test and was explained by the model. The proportion of PC variability was estimated by the R2 of the fit. ID was found to be correlated with PC1 and explained the largest part of the dataset variability (≥7 .37%, p-value = 3.71 × 10-9). To reduce the impact, ComBat28 was used to correct for facility ID effects.

[0406] Integration of repeated samples Data for 35 replicate samples between batches 1 and 2 of MMY2002 were collected for each tank. Protein concentrations were integrated by calculating the mean value.

[0407] Protein concentration difference analysis Responders vs. Non-responders DARZALEX™ (daratumumab) in responders vs. non-responders Statistical comparison of protein concentration distributions was performed using two complementary methods: (i) individual SOMA Wilcoxon rank sum test for mer (Hollander and Wolfe ,Ninparametirc Statistical Methods.New Y ork:John Wiley & Sons.1973.27~33(one-sam ple), 68–75 (two-sample), and (ii) all SOMAmers Limma analysis for Nucleic Acids (Ritchie, ME, et al., Acids Res.2015;20:43(7):e47) was used simultaneously to measure the standard All p values ​​are based on Benjamin E. Correction was performed using the mini-Hochberg (BH) method (Benjamini and Hochberg, (1995) JRStatist.Soc.B.57:289 ~300;R:A Language and Environment for St aistical Computing,R Development Core T eam,R Foundation for Statistical Computing ng,Vienna,Austria.2011;ISBN 3-900051-07- 0). The null hypothesis of no differential expression was rejected when the adjusted p-value was <0.05.

[0408] On-treatment vs. baseline Baseline versus treatment protein concentrations were analyzed using three different statistical methods: (i) two-way regression analysis; (ii) Wilcoxon signed-rank test, and (iii) Friedman test. Test (Johnson et al., (2007) Biostatistics 8 (1):118-127). All p values ​​are based on B for multiple hypothesis correction. The H method was used to correct for the FDR of the controls (Benjamini and Hochber g, JRStatist.Soc.B.57:289~300,1995). Treatment In addition to significance, two-way repeated measures analysis of variance (Chambers et al., Anal ysis of variance;designed experiments:Ch apter 5.Statistical Models in S,Editors JM Chambers and TJ Hastie.Wadsworth & Brookes / Cole. 1992) were used to determine the significant time point: the correlation between response classes was This was also applied to determine whether MAmers occurred. To specifically determine whether responders showed a different treatment effect, all subjects were Differences between on-treatment and baseline protein concentration values ​​were calculated and Wilcoxon rank sum tests were performed. A modified Wilcoxon rank sum test was applied as a post hoc test. Values ​​were corrected using the BH method, and the null hypothesis was rejected when the corrected p value was <0.05.

[0409] classifier training Using baseline protein concentration data from MMY2002, we developed a classifier to predict response. Four types of machine learning machines, Support Vector Machine (SVM), Random Forest Nested loop stratification using Regression Tree (RF), Naive Bayes (NB), and j48 decision trees The 10-fold cross-validation method was repeated 30 times. For each learner, the training procedure was balanced by 10. We started by creating a dataset with well-spaced partitions (outer loop). Keep one as the test cohort, while the remaining nine are the training cohort in the inner loop. In the inner loop, the training cohort was again split into 10 evenly balanced groups, and the inner training and We created inner test sets. We trained a learner on each of these inner training sets. This process was repeated 30 times for each cohort in the outer loop. The accuracy of each inner loop learner in predicting the set of features is used to select features and refine the model parameters. After completing 30 inner loops for each training cohort, Evaluate the performance of the outer loop (using optimized parameters and features) on the test cohort. The entire outer loop was then repeated 30 times to estimate all samples in the data set. The looping method yielded 30 different response predictions. The sensitivity and specificity statistics are compared for the final model trained on the full original dataset. were similar in how well they performed on new test cases.

[0410] Results from the MMY2002 trial Various comparisons were performed, including treatment-induced response-dependent changes in protein expression. One of the proteins that showed a decrease in expression over time in PD-L1 was PD PD-L1 protein expression increased over time in non-responders. Binding on T cells leads to a decrease in T cell function and an increase in Treg development. Figure 18 are the first and second cycles in responders, non-responders, and stable patients. Shown is the protein expression profile of PD-L1 at cycle 3.

[0411] PD-L1 binding to its receptor PD-1 suppresses antitumor responses and leads to T cell anergy and exhaustion. Without being bound to any particular theory, downregulation of PD-L1 by CD38 therapy may also result in improved enhancement of antitumor immune responses in solid tumors. The present invention may include the following aspects. [1] 1. A method of treating a patient having a solid tumor, comprising administering to a patient in need thereof a therapeutically effective amount of an antibody that specifically binds CD38 for a period of time sufficient to treat the solid tumor. [2] The method according to [1] above, wherein the antibody that specifically binds CD38 induces an immune response in the patient. [3] The method according to [2] above, wherein the immune response is an effector T cell (Teff) response. [4] The method according to [3] above, wherein the Teff response is mediated by CD4+ T cells or CD8+ T cells. [5] The method according to [4] above, wherein the Teff response is mediated by the CD8+ T cells. [6] The method according to [3] above, wherein the Teff response is an increase in the number of CD8+ T cells, an increase in CD8+ T cell proliferation, an increase in T cell clonal proliferation, an increase in CD8+ memory cell formation, an increase in antigen-dependent antibody production, an increase in cytokine production, an increase in chemokine production, or an increase in interleukin production (by cells). [7] The method according to [1] above, wherein the antibody that specifically binds to CD38 inhibits the function of immune suppressor cells. [8] The method according to [7] above, wherein the immune suppressor cells are regulatory T cells (Treg). [9] The method according to [8] above, wherein the Tregs are CD3+CD4+CD25+CD127dim T cells.

[10] The method according to [9] above, wherein the Treg expresses CD38.

[11] The method according to

[10] above, wherein the function of the Treg is inhibited by killing the Treg.

[12] The method according to

[11] above, wherein killing the Tregs is mediated by antibody-dependent cellular cytotoxicity (ADCC).

[13] The method according to [7] above, wherein the immune suppressor cells are myeloid-derived suppressor cells (MDSCs).

[14] The method according to

[13] above, wherein the MDSC is a CD11b+HLADR-CD14-CD33+CD15+ cell.

[15] The method according to

[14] above, wherein the CD11b+HLADR-CD14-CD33+CD15+ cells express CD38.

[16] The method according to

[15] above, wherein the function of the MDSC is inhibited by killing the MDSC.

[17] The method according to

[16] above, wherein the killing of MDSC is mediated by ADCC.

[18] The method according to [7] above, wherein the immune suppressor cells are regulatory B cells (Bregs).

[19] The method according to

[18] above, wherein the Bregs are CD19+CD24+CD38+ cells.

[20] The method according to

[19] above, wherein the function of the Breg is inhibited by killing the Breg. [twenty one] The method according to

[20] above, wherein the killing of Bregs is mediated by ADCC. [twenty two] The method according to [7] above, wherein the immune suppressor cells are present in bone marrow or peripheral blood. [twenty three] The method according to any one of the above-mentioned [1] to

[22] , wherein the solid tumor is melanoma, lung cancer, squamous non-small cell lung cancer (NSCLC), non-squamous NSCLC, colorectal cancer, prostate cancer, castration-resistant prostate cancer, stomach cancer, ovarian cancer, gastric cancer, liver cancer, pancreatic cancer, thyroid cancer, squamous cell carcinoma of the head and neck, carcinoma of the esophagus or gastrointestinal tract, breast cancer, fallopian tube cancer, brain cancer, urethral cancer, genitourinary cancer, endometriosis, cervical cancer, or a metastasis of the above cancers. [twenty four] The method according to any one of the above [1] to

[23] , wherein the solid tumor lacks detectable CD38 expression. [twenty five] The method according to any one of the above-mentioned [1] to

[24] , wherein the antibody that specifically binds to CD38 is a non-agonistic antibody.

[26] The method according to

[25] above, wherein the non-agonistic antibody induces proliferation of peripheral blood mononuclear cell samples in vitro in a statistically insignificant manner.

[27] The method according to any of the above-mentioned [1] to

[26] , wherein the antibody that specifically binds CD38 competes with an antibody comprising a heavy chain variable region (VH) of SEQ ID NO: 4 and a light chain variable region (VL) of SEQ ID NO: 5 for binding to CD38.

[28] The method according to

[27] above, wherein the antibody that specifically binds CD38 binds to at least the region SKRNIQFSCKNIYR (sequence number 2) and the region EKVQTLEAWVIHGG (sequence number 3) of human CD38 (sequence number 1).

[29] The method according to any of the above-mentioned [1] to

[28] , wherein the antibody that specifically binds CD38 comprises the amino acid sequences of heavy chain complementarity determining region (HCDR) 1, HCDR2, HCDR3, light chain complementarity determining region (LCDR) 1, LCDR2 and LCDR3 of SEQ ID NOs: 6, 7, 8, 9, 10 and 11, respectively.

[30] The method according to any of the above-mentioned [1] to

[29] , wherein the antibody that specifically binds CD38 comprises a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5.

[31] The antibody that specifically binds CD38 is a) a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15; b) VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c) a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19, or d) The method according to any one of the above-mentioned [1] to

[26] , comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of VH of SEQ ID NO: 20 and VL of SEQ ID NO: 21.

[32] The antibody that specifically binds CD38 is a) a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15; b) VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c) a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19, or d) The method according to

[31] above, comprising a VH of sequence number 20 and a VL of sequence number 21.

[33] The method according to any of the above-mentioned [1] to

[32] , wherein the antibody that specifically binds CD38 is administered in combination with a second therapeutic agent.

[34] The method according to

[33] above, wherein the second therapeutic agent is a chemotherapeutic agent, a cancer targeted therapy, a standard therapeutic agent for the treatment of solid tumors, or an immune checkpoint inhibitor.

[35] The method according to

[34] above, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-PD-L2 antibody, an anti-LAG3 antibody, an anti-TIM3 antibody, or an anti-CTLA-4 antibody.

[36] The method according to

[35] above, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody.

[37] the anti-PD-1 antibody a) a VH of SEQ ID NO: 22 and a VL of SEQ ID NO: 23, b) a VH of SEQ ID NO: 24 and a VL of SEQ ID NO: 25; c) a VH of SEQ ID NO: 32 and a VL of SEQ ID NO: 33, or d) The method according to

[36] above, comprising a VH of sequence number 34 and a VL of sequence number 35.

[38] The method according to

[35] above, wherein the immune checkpoint inhibitor is an anti-PD-L1 antibody.

[39] the anti-PD-L1 antibody a) a VH of SEQ ID NO: 26 and a VL of SEQ ID NO: 27, b) a VH of SEQ ID NO: 28 and a VL of SEQ ID NO: 29, or c) The method described in

[38] above, comprising a VH of sequence number 30 and a VL of sequence number 31.

[40] The method according to

[35] above, wherein the immune checkpoint inhibitor is an anti-PD-L2 antibody.

[41] The method according to

[35] above, wherein the immune checkpoint inhibitor is an anti-LAG3 antibody.

[42] The method according to

[35] above, wherein the immune checkpoint inhibitor is an anti-TIM-3 antibody.

[43] the anti-TIM-3 antibody a) a VH of SEQ ID NO: 36 and a VL of SEQ ID NO: 37, or b) The method described in

[42] above, comprising a VH of sequence number 38 and a VL of sequence number 39.

[44] The method according to any one of the above

[33] to

[43] , wherein the second therapeutic agent is administered simultaneously, sequentially, or separately.

[45] The method according to any of the above-mentioned [1] to

[44] , wherein the antibody that specifically binds to CD38 is administered intravenously.

[46] The method according to any of the above-mentioned [1] to

[44] , wherein the antibody that specifically binds CD38 is administered subcutaneously in a pharmaceutical composition comprising the antibody that specifically binds CD38 and hyaluronidase.

[47] The method according to

[46] above, wherein the hyaluronidase is rHuPH20 of SEQ ID NO: 40.

[48] The method according to any one of the above [1] to

[47] , wherein the patient is being treated or has already been treated with radiation therapy.

[49] The method according to any one of the above [1] to

[47] , wherein the patient has undergone or will undergo later surgery.

[50] A method for inhibiting the activity of immune suppressor cells, the method comprising contacting said immune suppressor cells with an antibody that specifically binds CD38.

[51] The method according to

[50] above, wherein the immune suppressor cells are Tregs.

[52] The method according to

[51] above, wherein the Tregs are CD3+CD4+CD25+CD127dim T cells.

[53] The method according to

[50] above, wherein the immune suppressor cells are MDSCs.

[54] The method according to

[53] above, wherein the MDSC is a CD11b+HLADR-CD14-CD33+CD15+ cell.

[55] The method according to

[50] above, wherein the immune suppressor cells are Bregs.

[56] The method according to

[55] above, wherein the Bregs are CD19+CD24+CD38+ cells.

[57] The method according to any one of the above-mentioned

[50] to

[56] , wherein the antibody that specifically binds CD38 is a non-agonistic antibody.

[58] The method according to

[57] above, wherein the non-agonistic antibody induces proliferation of peripheral blood mononuclear cell samples in vitro in a statistically insignificant manner.

[59] The method according to any of the above-mentioned

[50] to

[58] , wherein the antibody that specifically binds CD38 competes with an antibody comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5 for binding to CD38.

[60] The method according to

[59] above, wherein the antibody that specifically binds CD38 binds to at least the region SKRNIQFSCKNIYR (sequence number 2) and the region EKVQTLEAWVIHGG (sequence number 3) of human CD38 (sequence number 1).

[61] The method according to any of the above-mentioned

[50] to

[60] , wherein the antibody that specifically binds CD38 comprises the amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of SEQ ID NOs: 6, 7, 8, 9, 10 and 11, respectively.

[62] The method according to

[61] above, wherein the antibody that specifically binds CD38 comprises a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5.

[63] The antibody that specifically binds CD38 is a) a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15; b) VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c) a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19, or d) The method according to any one of the above-mentioned

[50] to

[58] , comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of VH of SEQ ID NO: 20 and VL of SEQ ID NO: 21.

[64] The antibody that specifically binds CD38 is a) a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15; b) VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c) a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19, or d) The method according to

[63] above, comprising a VH of sequence number 20 and a VL of sequence number 21.

[65] 1. A method of enhancing an immune response in a patient, comprising administering to the patient an antibody that specifically binds CD38.

[66] The method according to

[65] above, wherein the patient has cancer or a viral infection.

[67] The method according to

[65] or

[66] above, wherein the antibody that specifically binds CD38 is a non-agonistic antibody.

[68] The method according to

[67] above, wherein the non-agonistic antibody induces proliferation of peripheral blood mononuclear cell samples in vitro in a statistically insignificant manner.

[69] The method according to any of the above-mentioned

[65] to

[68] , wherein the antibody that specifically binds CD38 competes with an antibody comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5 for binding to CD38.

[70] The method according to any of the above-mentioned

[65] to

[69] , wherein the antibody that specifically binds CD38 binds to at least the region SKRNIQFSCIYR (sequence number 2) and the region EKVQTLEAWVIHGG (sequence number 3) of human CD38 (sequence number 1).

[71] The method according to any of the above-mentioned

[65] to

[70] , wherein the antibody that specifically binds CD38 comprises the amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of SEQ ID NOs: 6, 7, 8, 9, 10 and 11, respectively.

[72] The method according to any of the above-mentioned

[65] to

[70] , wherein the antibody that specifically binds CD38 comprises a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5.

[73] The antibody that specifically binds CD38 is a) a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15; b) VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c) a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19, or d) The method according to any one of the above

[65] to

[68] , comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of VH of SEQ ID NO: 20 and VL of SEQ ID NO: 21.

[74] The antibody that specifically binds CD38 is a) a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15; b) VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c) a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19, or d) The method according to

[73] above, comprising a VH of sequence number 20 and a VL of sequence number 21.

[75] 1. A method of treating a patient having a viral infection, comprising administering to said patient an antibody that specifically binds CD38 for a period of time sufficient to treat said viral infection.

[76] The method according to

[75] above, wherein the antibody that specifically binds CD38 is a non-agonistic antibody.

[77] The method according to

[75] or

[76] above, wherein the non-agonistic antibody induces proliferation of a peripheral blood mononuclear cell sample in vitro in a statistically insignificant manner.

[78] The method according to any of the above-mentioned

[75] to

[77] , wherein the antibody that specifically binds CD38 competes with an antibody comprising a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5 for binding to CD38.

[79] The method according to any of the above-mentioned

[75] to

[78] , wherein the antibody that specifically binds CD38 binds to at least the region SKRNIQFSCIYR (sequence number 2) and the region EKVQTLEAWVIHGG (sequence number 3) of human CD38 (sequence number 1).

[80] The method according to any of the above-mentioned

[75] to

[79] , wherein the antibody that specifically binds CD38 comprises the amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of SEQ ID NOs: 6, 7, 8, 9, 10 and 11, respectively.

[81] The method according to any of the above-mentioned

[75] to

[80] , wherein the antibody that specifically binds CD38 comprises a VH of SEQ ID NO: 4 and a VL of SEQ ID NO: 5.

[82] The antibody that specifically binds CD38 is a) a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15; b) VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c) a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19, or d) The method according to any one of the above

[75] to

[77] , comprising HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of VH of SEQ ID NO: 20 and VL of SEQ ID NO: 21.

[83] The antibody that specifically binds CD38 is a) a VH of SEQ ID NO: 14 and a VL of SEQ ID NO: 15; b) VH of SEQ ID NO: 16 and VL of SEQ ID NO: 17; c) a VH of SEQ ID NO: 18 and a VL of SEQ ID NO: 19, or d) The method according to

[82] above, comprising a VH of sequence number 20 and a VL of sequence number 21.

Claims

1. A pharmaceutical composition for use in suppressing immune suppressor cell activity in a patient with cancer, comprising: the pharmaceutical composition comprises an antibody that specifically binds CD38, the antibody is of the IgG1 isotype, The antibody is a) the heavy chain variable region (VH) amino acid sequence of SEQ ID NO:4, and b) the light chain variable region (VL) amino acid sequence of SEQ ID NO:5 13. A pharmaceutical composition comprising:

2. The pharmaceutical composition described in claim 1, wherein the cancer is multiple myeloma.

3. The pharmaceutical composition of claim 1, wherein the cancer is a solid tumor.

4. The pharmaceutical composition of claim 3, wherein the solid tumor is melanoma, lung cancer, squamous non-small cell lung cancer (NSCLC), non-squamous NSCLC, colorectal cancer, prostate cancer, castration-resistant prostate cancer, stomach cancer, ovarian cancer, gastric cancer, liver cancer, pancreatic cancer, thyroid cancer, squamous cell carcinoma of the head and neck, carcinoma of the esophagus or gastrointestinal tract, breast cancer, fallopian tube cancer, brain cancer, urethral cancer, genitourinary cancer, endometriosis, cervical cancer, or a metastasis of any of the above cancers.

5. The pharmaceutical composition described in any one of claims 1 to 4, wherein the immune suppressor cells include regulatory T cells (Treg).

6. a) the Treg comprises CD3 + CD4 + CD25 + CD127 dim T cells; b) the Tregs express CD38; c) the function of said Treg is inhibited by killing said Treg; or any combination thereof.

7. The pharmaceutical composition of claim 6, wherein the function of the Treg is inhibited by killing the Treg, and the killing of the Treg is mediated by antibody-dependent cellular cytotoxicity (ADCC).

8. The pharmaceutical composition described in any one of claims 1 to 4, wherein the immune suppressor cells include myeloid-derived suppressor cells (MDSCs).

9. The method of claim 8, wherein the MDSCs comprise CD11b + HLADR − CD14 − CD33 + CD15 + cells. b) the MDSCs express CD38; c) the function of the MDSCs is inhibited by killing the MDSCs; or any combination thereof.

10. The pharmaceutical composition of claim 9, wherein the function of the MDSCs is inhibited by killing the MDSCs, and the killing of the MDSCs is mediated by ADCC.

11. The pharmaceutical composition of any one of claims 1 to 4, wherein the immune suppressor cells include regulatory B cells (Breg).

12. The method of claim 11, wherein the Bregs comprise CD19+CD24+CD38+ cells. b) the function of the Breg is inhibited by killing the Breg; Or both a) and b).

13. The pharmaceutical composition of claim 12, wherein the function of the Breg is inhibited by killing the Breg, and the killing of the Breg is mediated by ADCC.

14. The pharmaceutical composition described in any one of claims 1 to 13, wherein the immune suppressor cells are present in bone marrow or peripheral blood.

15. The pharmaceutical composition of any one of claims 1 to 14, wherein the pharmaceutical composition is formulated for intravenous administration.

16. The pharmaceutical composition of any one of claims 1 to 14, further comprising hyaluronidase and formulated for subcutaneous administration.

17. A pharmaceutical composition described in any one of claims 1 to 16, wherein the patient is treated with an immune checkpoint inhibitor.

18. The pharmaceutical composition of claim 17, wherein the immune checkpoint inhibitor comprises an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-PD-L2 antibody, an anti-LAG3 antibody, an anti-TIM3 antibody, or an anti-CTLA-4 antibody.

19. The pharmaceutical composition of claim 18, wherein the immune checkpoint inhibitor comprises an anti-PD-1 antibody.

20. The anti-PD-1 antibody, a) a VH of SEQ ID NO: 22 and a VL of SEQ ID NO: 23; b) a VH of SEQ ID NO: 24 and a VL of SEQ ID NO: 25; c) a VH of SEQ ID NO: 32 and a VL of SEQ ID NO: 33; or 20. The pharmaceutical composition of claim 19, comprising d) a VH of SEQ ID NO: 34 and a VL of SEQ ID NO:

35.

21. The pharmaceutical composition of claim 18, wherein the immune checkpoint inhibitor comprises an anti-PD-L1 antibody.

22. The anti-PD-L1 antibody, a) a VH of SEQ ID NO: 26 and a VL of SEQ ID NO: 27; b) a VH of SEQ ID NO: 28 and a VL of SEQ ID NO: 29; or c) The pharmaceutical composition of claim 21, comprising a VH of SEQ ID NO: 30 and a VL of SEQ ID NO:

31.

23. The pharmaceutical composition of claim 18, wherein the immune checkpoint inhibitor comprises an anti-PD-L2 antibody.

24. The pharmaceutical composition of claim 18, wherein the immune checkpoint inhibitor comprises an anti-LAG3 antibody.

25. The pharmaceutical composition of claim 18, wherein the immune checkpoint inhibitor comprises an anti-TIM-3 antibody.

26. The anti-TIM-3 antibody, a) a VH of SEQ ID NO: 36 and a VL of SEQ ID NO: 37, or The pharmaceutical composition of claim 25, comprising a VH of SEQ ID NO: 38 and a VL of SEQ ID NO:

39.

27. ​​A pharmaceutical composition described in any one of claims 1 to 26, wherein the antibody that specifically binds CD38 comprises a heavy chain of SEQ ID NO: 12 and a light chain of SEQ ID NO: 13.

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