CD38 antibody and its use

Abstract

The present invention relates to an anti-CD38 antibody containing one or more mutations in the Fc region, and to the use of such an antibody in the treatment of a disease in a target, such as hematological malignancies.

Description

[Technical Field]

[0001] Field of Invention This invention relates to an anti-CD38 antibody containing one or more mutations in the Fc region, and to the use of such an antibody in the treatment of a disease in a subject. [Background technology]

[0002] Background of the Invention CD38 is a type II transmembrane glycoprotein typically found in hematopoietic cells and present at low levels in certain solid tissues. CD38 expression in hematopoietic cells is determined by the differentiation and activation state of the cells. Hematopoietic cells that have undergone differentiation lineage determination express the protein, but this is lost in mature cells and re-expressed in activated lymphocytes. CD38 is also expressed in B cells, with plasma cells expressing particularly high levels. Approximately 80% of quiescent NK cells and monocytes express CD38 at low levels, as do various other blood cell types, including lymph node germinal center lymphoblasts, intrafollicular cells, dendritic cells, erythrocytes, and platelets (Lee and Aarhus 1993; Zocchi, Franco et al. 1993; Malavasi, Funaro et al. 1994; Ramaschi, Torti et al. 1996). Regarding solid tissues, CD38 is expressed in the intestines by intraepithelial cells and lamina propria lymphocytes, in the brain by Purkinje cells and neurofibrillary tangles, in the prostate by epithelial cells, in the pancreas by β-cells, in bone by osteoclasts, in the eye by retinal cells, and in the muscular sheaths of smooth and striated muscle.

[0003] CD38 is expressed in a number of hematological malignancies. Expression has been observed in malignant cells, particularly in multiple myeloma (MM) (Lin, Owens et al. 2004) and chronic lymphocytic leukemia (CLL) (Damle 1999), and has also been reported in Waldenström macroglobulinemia (Konoplev, Medeiros et al. 2005), primary systemic amyloidosis (Perfetti, Bellotti et al. 1994), mantle cell lymphoma (Parry-Jones, Matutes et al. 2007), acute lymphoblastic leukemia (Keyhani, Huh et al. 2000), acute myeloid leukemia (Marinov, Koubek et al. 1993; Keyhani, Huh et al. 2000), NK cell leukemia (Suzuki, Suzumiya et al. 2004), NK / T cell lymphoma (Wang, Wang et al. 2015), and plasma cell leukemia (van de Donk, Lokhorst et al. 2012).

[0004] Other diseases in which CD38 expression may be involved include, for example, bronchial epithelial cancer of the lung, breast cancer (arising from malignant proliferation of the duct and lobular epithelium of the breast), pancreatic tumors (insulinomas) arising from β cells, tumors arising from the epithelium of the intestine (e.g., adenocarcinoma and squamous cell carcinoma), prostate cancer tumors, testicular seminomas, ovarian cancer, and neuroblastoma. Other disclosures also suggest a role for CD38 in autoimmune diseases such as Graves' disease, thyroiditis (Antonelli, Fallahi et al. 2001), type 1 and type 2 diabetes (Mallone and Perin 2006), and airway smooth muscle cell inflammation in asthma (Deshpande, White et al. 2005). Furthermore, CD38 expression is also associated with HIV infection (Kestens, Vanham et al. 1992; Ho, Hultin et al. 1993).

[0005] CD38 is a multifunctional protein. The functions attributed to CD38 include both receptor-mediated and (ecto)enzymatic activities in adhesion and signaling events. As an ectoenzyme, CD38 uses NAD + as a substrate for the formation of cyclic ADP-ribose (cADPR) and ADP-ribose (ADPR), but also for the formation of nicotinamide and nicotinic acid-adenine dinucleotide phosphate (NAADP). cADPR has been shown to function as a second messenger for Ca 2+ mobilization from the endoplasmic reticulum.

[0006] Several anti-CD38 antibodies have been described in the literature, for example, those described in WO 2006 / 099875 A1, WO2008037257 A2, WO 2011 / 154453 A1, WO 2007 / 042309 A1, WO 2008 / 047242 A1, WO2012 / 092612 A1; Cotner, Hemler et al. 1981; Ausiello, Urbani et al. 2000; Lande, Urbani et al. 2002; de Weers, Tai et al. 2011; Deckert, Wetzel et al. 2014; Raab, Goldschmidt et al. 2015; Eissler, Filosto et al. 2018; Roepcke, Plock et al. 2018; and Schooten 2018.

[0007] CD38 antibodies may affect CD38-expressing tumor cells through one or more of the following mechanisms: complement-dependent cell-mediated cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), programmed cell death, trogocytosis, elimination of immune suppressor cells, and regulation of enzyme activity (van de Donk, Janmaat et al. 2016; Krejcik, Casneuf et al. 2016; Krejcik, Frerichs et al. 2017; Chatterjee, Daenthanasanmak et al. 2018; van de Donk 2018). However, it was shown in 2014 that no CD38 antibody has been described that can not only effectively induce CDC, ADCC, and ADCP but also effectively inhibit CD38 enzyme activity (Lammerts van Bueren, Jakobs et al. 2014).

[0008] Optimizing effector function can improve the effectiveness of therapeutic antibodies for treating cancer or other diseases, for example, by improving the ability of antibodies to elicit an immune response against antigen-expressing cells. Such efforts are described, for example, in: WO 2013 / 004842 A2; WO 2014 / 108198 A1; WO 2018 / 031258 A1; Dall'Acqua, Cook et al. 2006; Moore, Chen et al. 2010; Desjarlais and Lazar 2011; Kaneko and Niwa 2011; Song, Myojo et al. 2014; Brezski and Georgiou 2016; Sondermann and Szymkowski 2016; Zhang, Armstrong et al. 2017; Wang, Mathieu et al. 2018.

[0009] WO 2020 / 012036, WO 2020 / 012038, and WO 2021 / 144457 describe advantageous anti-CD38 antibody variants with modified potency, as well as their uses and formulations (all are incorporated herein by reference).

[0010] While treatment regimens are currently available for CD38+ cancers such as MM, there is still a need for further treatment options because some patients are relapsed or refractory to currently available therapies.

[0011] Therefore, one object of the present invention is to provide means and methods for treating cancer by using the anti-CD38 antibody described herein, and to provide the anti-CD38 antibody described herein for use in the treatment of cancer, more specifically, hematological cancers such as MM.

[0012] For example, specific doses, ranges, and / or dosing regimens are provided that are advantageous with respect to such methods or uses for cancers that are known or identified as CD38-positive, such as MM. More specifically, specific doses, ranges, and / or dosing regimens that are advantageous with respect to the treatment of patients suffering from (relapsed or refractory) MM are provided herein. The dose ranges and / or dosing regimens provided herein have been evaluated as safe in human use and / or shown to be effective in the treatment of MM. [Overview of the Initiative]

[0013] The present invention relates to an anti-CD38 antibody having one or more mutations in the Fc region, particularly antibody C, and its use in the treatment of hematological malignancies such as MM. At least one of these mutations is in a residue corresponding to E430, E345, or S440 (amino acid residues are numbered according to the EU index) in the human IgG monohelic acid chain.

[0014] Therefore, in one aspect, the present invention provides a method for treating or preventing hematological malignancies, preferably multiple myeloma (MM), in subjects that need such treatment, preferably human subjects, comprising the step of administering an antibody that binds to human CD38 to the subject in a therapeutically effective amount. The antibody, a. An antigen-binding region comprising VH CDR1 having the sequence described in SEQ ID NO:2, VH CDR2 having the sequence described in SEQ ID NO:3, VH CDR3 having the sequence described in SEQ ID NO:4, VL CDR1 having the sequence described in SEQ ID NO:6, VL CDR2 having sequence AAS, and VL CDR3 having the sequence described in SEQ ID NO:7, and b. Fc region containing mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 (amino acid residues are numbered according to the EU index) in the human IgG monohelic acid. This includes methods.

[0015] In another aspect, the present invention provides a method for treating or preventing hematological malignancies, preferably multiple myeloma (MM), in subjects in need, preferably human subjects, comprising the step of administering an antibody or a pharmaceutical composition containing the antibody to the subject in a therapeutically effective amount. The antibody, a. A heavy chain comprising a VH region containing VH CDR1 having the sequence described in SEQ ID NO:2, VH CDR2 having the sequence described in SEQ ID NO:3, and VH CDR3 having the sequence described in SEQ ID NO:4, and a human IgG1 CH region having mutations in one or more of E430, E345, and S440 (amino acid residues are numbered according to the EU index); and b. A light chain comprising a VL region including VL CDR1 having the sequence described in SEQ ID NO:6, VL CDR2 having sequence AAS, and VL CDR3 having the sequence described in SEQ ID NO:7. This includes methods.

[0016] In another aspect, the present invention provides a method for treating or preventing hematological malignancies, preferably multiple myeloma (MM), in subjects in need, preferably human subjects, comprising the step of administering an antibody that binds to human CD38 to the subject in a therapeutically effective amount. The antibody, (a) A heavy chain comprising a VH region containing SEQ ID NO:1 and a human IgG1 CH region having mutations in one or more of E430, E345 and S440 (amino acid residues are numbered according to the EU index), and (b) Light chain containing VL including SEQ ID NO: 5 This includes methods.

[0017] In one embodiment, the mutation includes or consists of a mutation at position E430, preferably E430G.

[0018] In one embodiment, the antibody is administered in a dose of at least (approximately) 4 mg / kg body weight, for example, 4 mg / kg to (approximately) 24 mg / kg body weight.

[0019] In one embodiment, the antibody is administered in a dose of approximately 8 mg / kg to approximately 16 mg / kg body weight.

[0020] In one embodiment, the antibody is administered at a dose of approximately 16 mg / kg body weight.

[0021] In one embodiment, the antibody is administered in a 28-day (4-week) cycle, with weekly administration in cycles 1 and 2 (Q1W), bi-weekly administration in cycles 3-6 (Q2W), and monthly administration from cycle 7 onward (Q4W), with the initial dose being a divided dose.

[0022] In one embodiment, hematological malignancies are cancers that are refractory to previous therapies, such as those involving anti-CD38 antibodies, such as daratumumab or isatuximab.

[0023] In one embodiment, the hematological malignancy is (relapsed or refractory) multiple myeloma or (relapsed or refractory) DLBCL.

[0024] In one embodiment, administration of the antibody induces one or more therapeutic effects (compared to baseline). In another embodiment, administration of the antibody improves one or more therapeutic effects (compared to baseline).

[0025] The aforementioned one or more therapeutic effects may be the overall response rate, duration of response, and time to response.

[0026] The one or more therapeutic effects described above may be a strict complete response, a complete response, a best partial response, a partial response, a minimal response, or a stable state.

[0027] In one embodiment, the one or more therapeutic effects are improved compared to those of a reference antibody.

[0028] In one embodiment, the subject may be treated for the management of neutropenia or an infusion reaction.

[0029] In one embodiment, the target exhibits faster clearance of the antibody compared to the reference antibody, or the antibody exhibits faster clearance compared to the reference antibody.

[0030] In one embodiment, the administration of the antibody has the following effects: a. Activation of the complement system in the aforementioned object; b. Depletion of peripheral blood NK cells in the subject; c. Expansion and proliferation of peripheral blood T cells in the aforementioned subjects It may have one or more of these.

[0031] In one embodiment, the induction of complement activation is preferably greater than that of a reference antibody.

[0032] In one embodiment, the administration of the antibody does not result in a (substantial) dose-dependent increase in plasma levels of pro-inflammatory cytokines.

[0033] In one embodiment, administration of the antibody does not induce a dose-dependent reduction of B cells, T cells, monocytes, and / or NKT-like cells.

[0034] In one embodiment, the reference antibody does not contain mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440, and preferably the reference antibody is an IgG1 antibody, such as a wild-type IgG1 antibody. The reference antibody may be daratumumab or isatuximab.

[0035] In one embodiment, the antibody is a) The antibody at a concentration of 1 to 200 mg / mL, optionally; b) 5-40 mM histidine or acetate; c) 100-400 mM sorbitol or sucrose; and d) Surfactants It is included in compositions containing the following:

[0036] In one embodiment, the antibody is It has a pH of approximately 6, and Optionally, in aqueous solution state, a) The antibody at approximately 20 mg / mL, b) Histidine at approximately 20 mM, c) Approximately 250 mM sorbitol, and d) Polysorbate 80 at approximately 0.04% w / v To include, consist of, or essentially be made from It is included in compositions containing the following:

[0037] In one aspect, the present invention relates to an anti-CD38 antibody, or a composition comprising such antibody, in any aspect or embodiment as described herein, for use in treating or preventing hematological malignancies involving cells expressing CD38 as described herein.

[0038] In one aspect, the present invention relates to an anti-CD38 antibody, or a composition comprising such antibody, in any aspect or embodiment as described herein, for use in treating or preventing hematological malignancies in a subject comprising cells expressing human CD38 as described herein.

[0039] In one aspect, the present invention provides an anti-CD38 antibody, or a composition comprising the antibody, in any of the embodiments and aspects described herein, for use in the prevention or treatment of hematological malignancies described herein.

[0040] In one aspect, the present invention relates to an anti-CD38 antibody, or a composition comprising the antibody, in any aspect or embodiment described herein, for use as a pharmaceutical for treating or preventing hematological malignancies as described herein.

[0041] In one aspect, the present invention provides an anti-CD38 antibody, or a composition comprising the antibody, in any of the embodiments and aspects described herein, for the manufacture of a pharmacopoeia for the prevention or treatment of hematological malignancies described herein.

[0042] These and other aspects and embodiments of the present invention will be described in more detail below. [Brief explanation of the drawing]

[0043] [Figure 1]The amino acid sequence alignments using Clustal 2.1 software are shown for the human IgG1m(a), IgG1m(f), IgG2, IgG3, and IgG4 Fc segments corresponding to residues P247-K447 in the human IgG1 heavy chain, where amino acid residues are numbered according to the EU index described in Kabat. The amino acid sequences shown correspond to residues 130-330 of the heavy chain constant region of human IgG1 allotype variants designated as IgG1m(za) (SEQ ID NO:64; UniProt accession number P01857), IgG1m(f) (SEQ ID NO:65), IgG1m(z) (SEQ ID NO:66), IgG1m(a) (SEQ ID NO:67), and IgG1m(x) (SEQ ID NO:68); residues 126-326 of the IgG2 heavy chain constant region (SEQ ID NO:79; UniProt accession number P01859); residues 177-377 of the IgG3 heavy chain constant region (SEQ ID NO:80; UniProt accession number P01860); and residues 127-327 of the IgG4 heavy chain constant region (SEQ ID NO:81; UniProt accession number P01861). [Figure 2] A schematic diagram of the IgG-C-E430G clinical trial design is shown. DL = dose level; IA = interim analysis; IV = intravenous administration; MTD = maximum tolerated dose; RP2D = recommended dose for phase 2; RRMM = relapsed or refractory multiple myeloma; R / R DLBCL = relapsed or refractory diffuse large B-cell lymphoma. [Figure 3] This graph shows the absolute NK cell count (CD3- / CD56+ / CD16+ cells / μL) in the peripheral blood of RRMM patients treated with IgG1-C-E430G during the dose-escalation phase of the IgG-C-E430G clinical trial. NK cell counts are shown over time and grouped by dose cohort (0.2 / 0.6 mg / kg, 2 mg / kg, 4 mg / kg, 8 mg / kg, 16 mg / kg, and 24 mg / kg). Horizontal dashed lines marked with an asterisk (*) indicate 5 cells / μL, and vertical dotted lines at C1D1, C1D8, C1D15, and C1D21 indicate administration of IgG1-C-E430G. [Figure 4]This graph shows the relative change in NK cell count (% CD3- / CD56+ / CD16+ cells) from C1D1 in peripheral blood patients with RRMM treated with IgG1-C-E430G during the dose-escalation phase of the IgG-C-E430G clinical trial. The relative change in NK cell count is shown over time and grouped by dose cohort (0.2 / 0.6 mg / kg, 2 mg / kg, 4 mg / kg, 8 mg / kg, 16 mg / kg, and 24 mg / kg). The horizontal dashed line marked with an asterisk (*) indicates the relative NK cell count at C1D1 (baseline level), and the vertical dotted lines at C1D1, C1D8, C1D15, and C1D21 indicate administration of IgG1-C-E430G. [Figure 5] This graph shows the absolute T cell count (CD3+ cells / μL) in the peripheral blood of RRMM patients treated with IgG1-C-E430G during the dose-escalation phase of the IgG-C-E430G clinical trial. T cell counts are shown over time and grouped by dose cohort (0.2 / 0.6 mg / kg, 2 mg / kg, 4 mg / kg, 8 mg / kg, 16 mg / kg, and 24 mg / kg). Horizontal dashed lines marked with an asterisk (*) indicate 5 cells / μL, and vertical dotted lines at C1D1, C1D8, C1D15, and C1D21 indicate administration of IgG1-C-E430G. [Figure 6] This graph shows the relative change in T cell count (% CD3+ cells) from C1D1 in peripheral blood of RRMM patients treated with IgG1-C-E430G during the dose-escalation phase of the IgG-C-E430G clinical trial. The relative change in T cell count is shown over time and grouped by dose cohort (0.2 / 0.6 mg / kg, 2 mg / kg, 4 mg / kg, 8 mg / kg, 16 mg / kg, and 24 mg / kg). The horizontal dashed line marked with an asterisk (*) indicates the relative T cell count at C1D1 (baseline level), and the vertical dotted lines at C1D1, C1D8, C1D15, and C1D21 indicate IgG1-C-E430G administration. [Figure 7]This graph shows the absolute cytokine concentrations in the peripheral blood of RRMM patients treated with IgG1-C-E430G during the dose-escalation phase of the IgG-C-E430G clinical trial. Levels of interferon-gamma (IFNγ), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and tumor necrosis factor α (TNFα) are shown over time and grouped by dose cohort (0.2 / 0.6 mg / kg, 2 mg / kg, 4 mg / kg, 8 mg / kg, 16 mg / kg, and 24 mg / kg). The vertical dotted lines at C1D1, C1D8, C1D15, and C1D21 indicate administration of IgG1-C-E430G. Subjects 0016, 0018, and 0024 had out-of-scale values ​​for parameter IL-8, and subject 0019 had out-of-scale values ​​for parameter INFγ. [Figure 8] A schematic diagram of the bioanalysis method is shown. IgG1-C-E430G (intermediate gray antibody) was captured using an anti-idiotype antibody (dark gray - lower antibody) as a coating reagent. The captured drug was detected with a sulfo-tagged anti-idiotype secondary antibody (light gray - upper antibody). The signal was quantified by electrochemiluminescence. [Figure 9] This graph shows serum concentrations of IgG1-C-E430G (μg / mL) grouped by dose level, plotted against time (days) on a semi-logarithmic scale. Symbols represent individual observed serum concentrations; lines connect the observed values ​​for each patient. The actual time of sample collection was used for pre-treatment samples. Vertical dashed lines represent the planned time of drug administration. Horizontal lines represent the limit of quantification in the 0.05 μg / mL bioanalysis assay. Observed values ​​below the limit of quantification were plotted at half the limit of quantification (0.025 μg / mL). [Figure 10]The AUC0-t(d*μg / mL) and CL(L / d / kg) relative to dose (mg / kg) are shown for each patient, calculated where possible, on a log-log scale from the PK profiles collected per dose on days 1 / 2 of cycle 1, days 8 / 9 of cycle 1 (patient E only), and day 1 of cycle 2. The symbols and error bars represent the mean and standard deviation for each dose and PK profile. [Figure 11A] This panel shows absolute and relative CD4+ T cell counts (CD3+ / CD4+ cells) in peripheral blood of RRMM patients treated with IgG1-C-E430G during the dose-escalation phase of the first-in-human clinical trial (data cutoff: August 14, 2023). Panel A shows absolute CD4+ T cell counts (CD3+ / CD4+ cells / μL). Panel B shows the relative change in CD4+ T cell counts (% CD3+ / CD4+ cells) compared to CD4+ T cell counts in peripheral blood C1D1. CD4+ T cell counts and relative changes in CD4+ T cell counts are shown over time and grouped by dose cohort (0.2 / 0.6 mg / kg, 2 mg / kg, 4 mg / kg, 8 mg / kg, 16 mg / kg, and 24 mg / kg). The horizontal dashed line in Panel B indicates a 50% increase in CD3+ / CD4+ cells compared to baseline. The maximum value for the y-axis of panel B is set to 150%. Values ​​exceeding this limit are permitted. [Figure 11B] Refer to the explanation in Figure 11A. [Figure 12A]This panel shows absolute and relative CD8+ T cell counts (CD3+ / CD8+ cells) in peripheral blood of RRMM patients treated with IgG1-C-E430G during the dose-escalation phase of the first-in-human clinical trial (data cutoff: August 14, 2023). Panel A shows absolute CD8+ T cell counts (CD3+ / CD8+ cells / μL). Panel B shows the relative change in CD8+ T cell counts (% CD3+ / CD8+ cells) compared to CD8+ T cell counts in peripheral blood C1D1. CD8+ T cell counts and relative changes in CD8+ T cell counts are shown over time and grouped by dose cohort (0.2 / 0.6 mg / kg, 2 mg / kg, 4 mg / kg, 8 mg / kg, 16 mg / kg, and 24 mg / kg). The horizontal dashed line in Panel B indicates a 50% increase in CD3+CD8+ cells compared to baseline. The maximum value for the y-axis of panel B is set to 150%. Values ​​exceeding this limit are permitted. [Figure 12B] Refer to the explanation in Figure 12A. [Figure 13A] This panel shows the absolute and relative NK cell counts (CD3- / CD56+ / CD16+) over time in peripheral blood patients with RRMM who received 16 mg / kg of IgG1-C-E430G during Expansion Part A of the First-in-Human Clinical Trial. Panel A shows the absolute NK cell count (CD3- / CD56+ / CD16+ cells / μL). Panel B shows the relative change in NK cell count (% CD3- / CD56+ / CD16+ cells) compared to the NK cell count at C1D1 in peripheral blood. The maximum value on the y-axis of Panel B is set to 150%. Values ​​exceeding this limit may exist. [Figure 13B] Refer to the explanation in Figure 13A. [Figure 14A]This panel shows the absolute and relative CD4+ T cell counts (CD3+ / CD4+ cells) in peripheral blood of RRMM patients administered 16 mg / kg IgG1-C-E430G during Expansion Part A of the First-in-Human Clinical Trial (data cutoff: August 14, 2023). Panel A shows the absolute CD4+ T cell count (CD3+ / CD4+ cells / μL). Panel B shows the relative change in CD4+ T cell count (% CD3+ / CD4+ cells) compared to the CD4+ T cell count at C1D1 in peripheral blood. The horizontal dashed line in Panel B indicates a 50% increase in CD3+ / CD4+ cells compared to baseline. The maximum value on the y-axis in Panel B is set at 150%. Values ​​may exist beyond this limit. [Figure 14B] Refer to the explanation in Figure 14A. [Figure 15A] This panel shows the absolute and relative CD8+ T cell counts (CD3+ / CD8+ cells) in peripheral blood of RRMM patients administered 16 mg / kg IgG1-C-E430G during Expansion Part A of the First-in-Human Clinical Trial (data cutoff: August 14, 2023). Panel A shows the absolute CD8+ T cell count (CD3+ / CD8+ cells / μL). Panel B shows the relative change in CD8+ T cell count (% CD3+ / CD8+ cells) compared to the CD8+ T cell count at C1D1 in peripheral blood. The horizontal dashed line in Panel B indicates a 50% increase in CD3+ / CD8+ cells compared to baseline. The maximum value on the y-axis in Panel B is set at 150%. Values ​​may exist beyond this limit. [Figure 15B] Refer to the explanation in Figure 15A. [Figure 16] This shows the absolute cytokine concentrations in the peripheral blood of RRMM patients who received 16 mg / kg of IgG1-C-E430G during the expanded Part A of the First-in-Human Clinical Trial (data cutoff: August 14, 2023). It also shows the levels of interferon-gamma (IFNγ), interleukin-10 (IL-10), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNFα) over time. [Figure 17]This shows a comparison of serum concentrations of IgG1-C-E430G in RRMM patients (A) who received a dose of 16 mg / kg during the dose escalation study of the First-in-Human Clinical Trial, and in RRMM patients (B) who received a dose of 16 mg / kg during the expanded Part A. [Modes for carrying out the invention]

[0044] Detailed description of the invention Certain terms are used for clarity when describing aspects of the present invention. However, the present invention is not intended to be limited to the specific terms thus selected, and it is understood that each specific term includes all technical equivalents that operate in a similar manner to achieve a similar purpose.

[0045] definition As used herein, the term "CD38" generally refers to human CD38 having the sequence described in SEQ ID NO:38 (UniProtKB - P28907 (CD38_HUMAN)), but may also refer to its variants, isoforms, and orthologues unless inconsistent with the context. Variants of human CD38 with S274, Q272R, T237A, or D202G mutations are described in WO 2006 / 099875 A1 and WO 2011 / 154453 A1.

[0046] The term "immunoglobulin" refers to a class of structurally related glycoproteins consisting of two pairs of polypeptide chains: one pair of low-molecular-weight light (L) chains and one pair of heavy (H) chains, all four chains potentially interconnected by disulfide bonds. The structure of immunoglobulins is well characterized; see, for example, Chapter 7 of Fundamental Immunology (Paul, W., ed., 2nd edition, Raven Press, NY (1989)). Briefly, each heavy chain typically consists of a variable (VH) region and a constant (CH) region. The CH region typically consists of three domains: CH1, CH2, and CH3. The heavy chains are typically interconnected via disulfide bonds in a so-called "hinge region." Each light chain typically consists of a variable (VL) region and a constant region, the latter typically consisting of one domain, CL. The VH and VL regions can be further subdivided into hypervariable regions, also called complementarity-determining regions (CDRs), which are sandwiched between more conserved regions called framework regions (FRs). Each VH and VL region typically consists of three CDRs and four FRs, arranged in the following order from amino-terminus to carboxy-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 (see also Chothia and Lesk J. Mol. Biol. 196, 901 917 (1987)).

[0047] Unless otherwise specified or unless inconsistent with the context, CDR sequences herein are identified according to the IMGT rules using DomainGapAlign (Lefranc MP., Nucleic Acids Research 1999;27:209-212 and Ehrenmann F., Kaas Q. and Lefranc M.-P. Nucleic Acids Res., 38, D301-307 (2010); see also internet address www.imgt.org / ).

[0048] Unless otherwise specified or inconsistent with the context, references to amino acid positions of CH or Fc regions / domains in this invention follow EU numbering (Edelman et al., Proc Natl Acad Sci US A. 1969 May;63(1):78-85; Kabat et al., Sequences of proteins of immunological interest. 5th edition - 1991 NIH publication number 91-3242). However, amino acid residues of CH of isotypes other than human IgG1 may alternatively be referred to by the corresponding amino acid positions in the wild-type human IgG1 heavy chain, where the amino acid residues are numbered according to the EU index. Specifically, the corresponding amino acid positions can be identified as shown in Figure 1, i.e., by (a) aligning the amino acid sequence of a non-IgG1 constant region (or segment) with the amino acid sequence of a human IgG1 heavy chain (or segment), where the amino acid residues are numbered according to the EU index, and (b) identifying which amino acid position in the IgG1 heavy chain that the amino acid residue aligns to. Therefore, the location of such an amino acid residue may be referred to herein as “the amino acid residue at the corresponding position,” followed by the amino acid position of the wild-type human IgG1 heavy chain, numbered according to the EU index. When referring to one or more of several different amino acid positions, this may be referred herein as “a mutation of one or more amino acid residues at a position selected from the group consisting of the corresponding positions,” “a mutation of one or more amino acid residues at the corresponding position,” or simply “a mutation of one or more amino acid residues selected from the group consisting of the corresponding positions,” followed by two or more amino acid positions of the human wild-type IgG1 heavy chain (e.g., E430, E345, and S440), where the amino acid residues are numbered according to the EU index.

[0049] As used herein, the term “hinge region” is intended to refer to the hinge region of an immunoglobulin heavy chain. Therefore, for example, the hinge region of a human IgG1 antibody corresponds to amino acids 216-230 according to EU numbering.

[0050] As used herein, the terms “CH2 region” or “CH2 domain” are intended to refer to the CH2 region of an immunoglobulin heavy chain. Therefore, for example, the CH2 region of a human IgG1 antibody corresponds to amino acids 231-340 according to EU numbering. However, the CH2 region may also be any of the other subtypes described herein.

[0051] As used herein, the terms “CH3 region” or “CH3 domain” are intended to refer to the CH3 region of an immunoglobulin heavy chain. Therefore, for example, the CH3 region of a human IgG1 antibody corresponds to amino acids 341-447 according to EU numbering. However, the CH3 region may also be any of the other subtypes described herein.

[0052] 2 (WO2008 / 003116), and dual scFv-fusions. The term antibody should be understood to include, unless otherwise specified: monoclonal antibodies (e.g., human monoclonal antibodies), polyclonal antibodies, chimeric antibodies, humanized antibodies, monospecific antibodies (e.g., bivalent monospecific antibodies), bispecific antibodies, antibodies of any isotype and / or allotype; antibody mixtures (recombinant polyclonals), multimeric Fc proteins as described in WO2015 / 158867, and fusion proteins as described in WO2014 / 031646, produced by techniques (Oligoclonics) developed by Symphogen and Merus. While these various antibody fragments and formats are generally included in the meaning of antibody, they are collectively and independently unique features of the present invention, exhibiting different biological properties and utility.

[0053] The terms "CD38 antibody" or "anti-CD38 antibody" used herein refer to antibodies that specifically bind to the antigen CD38.

[0054] As used herein, the term "human antibody" is intended to include antibodies having a variable region and a constant region derived from a human germline immunoglobulin sequence. The human antibodies of the present invention may include amino acid residues not encoded by the human germline immunoglobulin sequence (e.g., mutations, insertions, or deletions introduced by random or site-directed mutagenesis in vitro, or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which a CDR sequence derived from the germline of another mammalian species, such as mouse, is grafted onto a human framework sequence.

[0055] As used herein, terms such as "monoclonal antibody," "monoclonal Ab," "monoclonal antibody composition," and "mAb" refer to preparations of a single-molecule Ab molecule. Monoclonal antibody compositions exhibit a single binding specificity and affinity for a specific epitope. Therefore, the term "human monoclonal antibody" refers to an Ab exhibiting single binding specificity, having a variable region and a constant region derived from a human germline immunoglobulin sequence. Human mAbs can be produced from hybridomas, which include B cells obtained from transgenic or transchromosomal non-human animals, such as transgenic mice, having a genome containing a human heavy-chain transgene repertoire and a light-chain transgene repertoire rearranged to produce functional human antibodies, fused to immortalized cells.

[0056] As used herein, “isotype” refers to an immunoglobulin class encoded by a heavy chain constant region gene, including, for example, IgG1, IgG2, IgG3, IgG4, IgD, IgA1, IgA2, IgE, and IgM, as well as any allotypes thereof, such as IgG1m(z), IgG1m(a), IgG1m(x), IgG1m(f), etc., and mixed allotypes thereof, such as IgG1m(za), IgG1m(zax), IgG1m(fa), etc. (see, for example, de Lange, Experimental and Clinical Immunogenetics 1989;6(1):7-17).

[0057] Furthermore, each heavy chain isotype can be combined with either a kappa(k) or lambda(l) light chain. As used herein, the term “mixed isotype” refers to the Fc region of an immunoglobulin produced by combining the structural features of one isotype with a similar region from another isotype, thereby generating a hybrid isotype. A mixed isotype includes an Fc region having a sequence composed of two or more isotypes selected from the following: IgG1, IgG2, IgG3, IgG4, IgD, IgA1, IgGA2, IgE, or IgM, thereby generating combinations such as IgG1 / IgG3, IgG1 / IgG4, IgG2 / IgG3, IgG2 / IgG4, or IgG1 / IgA.

[0058] As used herein, the term "full-length antibody" refers to an antibody (e.g., parent antibody or variant antibody) that contains all constant and variable heavy and light chain domains, corresponding to those typically found in the wild-type antibody of the isotype.

[0059] As used herein, "full-length bivalent monospecific monoclonal antibody" refers to a bivalent monospecific antibody (e.g., parent antibody or variant antibody) formed by one pair of identical HCs and one pair of identical LCs, where the constant domain and variable domain correspond to those typically found in antibodies of a particular isotype.

[0060] As used herein, the terms “antigen-binding region,” “binding region,” or “antigen-binding domain” refer to a region of an antibody that can bind to an antigen. This binding region is typically defined by the VH and VL domains of an antibody, which can be further subdivided into hypervariable regions, also called complementarity-determining regions (CDRs), sandwiched between more conserved regions called framework regions (FRs). These hypervariable regions may have hypervariability in the morphology of sequenced and / or structurally defined loops. The antigen can be any molecule, such as a polypeptide present on a cell.

[0061] As used herein, the term "target" refers to the molecule to which the antigen-binding domain of an antibody binds. A target includes any antigen against the produced antibody. The terms "antigen" and "target" are interchangeable in relation to the antibody and may constitute the same meaning and purpose with respect to any aspect or embodiment of the present invention.

[0062] The term "epitope" refers to a protein determinant that can specifically bind to an antibody variable domain. Epitopes typically consist of surface groupings of molecules, such as amino acids, sugar side chains, or combinations thereof, and usually possess specific three-dimensional structural and charge properties. Constructive and non-constructive epitopes are distinguished in that binding to the former is lost in the presence of a denaturing solvent, while binding to the latter is not. Epitopes may contain amino acid residues directly involved in binding (also called the immunodominant component of the epitope) and other amino acid residues that are not directly involved in binding.

[0063] As used herein, “variant” refers to a protein sequence or polypeptide sequence in which one or more amino acid residues differ from the parent sequence or reference sequence. A variant may have, for example, at least 80%, for example, 90%, 95%, 97%, 98%, or 99% sequence identity with respect to the parent sequence or reference sequence. Furthermore or alternatively, a variant may differ from the parent sequence or reference sequence by mutations, e.g., substitutions, insertions, or deletions of 12 or fewer amino acid residues, for example, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue. Accordingly, “variant antibody” or “antibody variant” is used interchangeably herein and refers to an antibody in which one or more amino acid residues differ from the parent antibody or reference antibody, for example, in the antigen-binding region, the Fc region, or both. Similarly, a “variant Fc region” or “Fc region variant” refers to an Fc region that differs from the parent or reference Fc region amino acid sequence by one or more amino acid residues, optionally 12 or fewer, e.g., 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue mutations, e.g., substitutions, insertions, or deletions. The parent or reference Fc region is typically the Fc region of a human wild-type antibody and, depending on the context, may be a specific isotype. In dimerized form, a variant Fc region can be homodimer or heterodimer, for example, one amino acid sequence of the dimerized Fc region contains mutations while the other is identical to the parent or reference wild-type amino acid sequence. Examples of wild-type (typically parent or reference) IgG CH and variant IgG constant region amino acid sequences containing Fc region amino acid sequences are shown in Table 4.

[0064] In the context of the present invention, a conservative substitution may be defined as a substitution within the following class of amino acids: - Acidic residues: Asp(D) and Glu(E) - Basic residues: Lys(K), Arg(R), and His(H) - Hydrophilic uncharged residues: Ser(S), Thr(T), Asn(N), and Gln(Q) - Aliphatic uncharged residues: Gly(G), Ala(A), Val(V), Leu(L), and Ile(I) - Nonpolar, uncharged residues: Cys(C), Met(M), and Pro(P) - Aromatic residues: Phe(F), Tyr(Y), and Trp(W).

[0065] Alternative conserved amino acid residue substitution classes: 1. AST 2. DE 3. NQ 4. RK 5. ILM 6. FYW.

[0066] Physical and functional classification of amino acid residue alternatives: - Alcohol group-containing residues: S and T - Aliphatic residues: I, L, V, and M - Cyclo-alkenyl-related residues: F, H, W, and Y - Hydrophobic residues: A, C, F, G, H, I, L, M, R, T, V, W, and Y - Negatively charged residues: D and E - Polar residues: C, D, E, H, K, N, Q, R, S, and T - Positively charged residues: H, K, and R - Small residues: A, C, D, G, N, P, S, T, and V - Very small residues: A, G, and S - Residues involved in turn formation: A, C, D, E, G, H, K, N, Q, R, S, P, and T - Movable residues: Q, T, K, S, G, N, D, E, and R.

[0067] As used herein, “sequence identity” refers to the percentage identity between two sequences as a function of the number of identical positions shared by the two sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap (i.e., homology percentage = number of identical positions / total number of positions × 100). The percentage identity between two nucleotide or amino acid sequences can be determined, for example, using the algorithm of E. Meyers and W. Miller, Comput. Appl. Biosci 4, 11-17 (1988), which is incorporated into the ALIGN program (version 2.0), using a PAM 120 weight residue table, gap length penalty 12, and gap penalty 4. Furthermore, the percentage identity between two amino acid sequences can be determined using the algorithm of Needleman and Wunsch, J. Mol. Biol. 48, 444-453 (1970). Other tools for sequence alignment are available on the internet and include, but are not limited to, Clustal Omega and EMBOSS Needle on the EMBL-EBI website www.ebi.ac.uk. Typically, default settings may be used.

[0068] In the context of this invention, unless otherwise specified, the following notation is used to describe mutations: the name of the mutated amino acid, followed by the position number of the mutation, and then the content that the mutation encompasses. Therefore, if the mutation is a substitution, the name of the amino acid that replaces the previous amino acid is included, and if an amino acid is deleted, " * This is indicated by ", and if the mutation is addition, the added amino acid is placed after the original amino acid. The amino acid name may be a one-letter code or a three-letter code. Therefore, for example, substitution of glutamic acid at position 430 with glycine is indicated as E430G, substitution of glutamic acid at position 430 with any amino acid is indicated as E430X, and deletion of glutamic acid at position 430 is indicated as E430 *It is shown as, and the addition of proline after the glutamic acid at position E430 is shown as E430EP.

[0069] As used herein, "immune-suppressive cells" refers to immune cells that can suppress the immune response in a subject, for example, by suppressing the activity of effector T cells and / or inhibiting the proliferation of T cells. Examples of such immune-suppressive cells include, but are not limited to, regulatory T cells (Tregs), regulatory B cells (Bregs), and myeloid-derived suppressor cells (MDSCs). Furthermore, immunosuppressive NK cells, NKT cells, macrophages, and antigen-presenting cells (APCs) also exist. An example of the phenotype of immunosuppressive NK cells is CD56 bright CD16 - and.

[0070] "Regulatory T cells" or "Tregs" or "Treg" refers to T lymphocytes that regulate the activity of other T cells and / or other immune cells, usually by suppressing their activity. An example of the Treg phenotype is CD3 + CD4 + CD25 + CD127 dim and. Tregs may further express Foxp3. It should be understood that Tregs cannot be completely limited to this phenotype.

[0071] "Effector T cells" or "Teffs" or "Teff" refers to T lymphocytes that perform the function of an immune response, for example, killing tumor cells and / or activating an anti-tumor immune response that results in the clearance of tumor cells from the body. Examples of the Teff phenotype include CD3 + CD4 + and CD3 + CD8 + are included. Teffs may secrete, contain, or express markers such as IFNγ, granzyme B, ICOS, etc. It should be understood that Teffs cannot be completely limited to these phenotypes.

[0072] "Bone marrow-derived suppressor cells," or "MDSCs," or simply "MDSCs" refers to a specific population of hematopoietic cells that express the macrophage / monocyte marker CD11b and the granulocyte marker Gr-1 / Ly-6G. An example of an MDSC phenotype is CD11b + HLA-DR - CD14 - CD33 + CD15 + MDSCs also typically exhibit low or undetectable expression of the mature antigen-presenting cell markers MHC class II and F480. MDSCs are immature cells of the myeloid lineage and can further differentiate into other cell types such as macrophages, neutrophils, dendritic cells, monocytes, and granulocytes. MDSCs can be found spontaneously in the normal adult bone marrow of humans and animals, or in normal hematopoietic sites such as the spleen.

[0073] "Regulatory B cells," or "Bregs," refer to B lymphocytes that suppress the immune response. An example of the Breg phenotype is CD19 + CD24 + CD38 + Breg can suppress the immune response by inhibiting T cell proliferation mediated by IL-10 secreted by Breg. It should be understood that other Breg subsets exist, for example, as described in Ding et al., (2015) Human Immunology 76: 615-621.

[0074] As used herein, the term “effector cell” refers to an immune cell involved in the effector phase of the immune response. Exemplary immune cells include cells of myeloid or lymphoid origin, such as lymphocytes (including B cells and T cells, including cytolytic T cells (CTLs)), killer cells, natural killer cells, macrophages, monocytes, eosinophils, polymorphonuclear cells, such as neutrophils, granulocytes, mast cells, and basophils. Some effector cells express Fc receptors (FcRs) or complement receptors to perform specific immune functions. In some embodiments, effector cells, such as natural killer cells, can induce ADCC. For example, monocytes, macrophages, neutrophils, dendritic cells, and Kupffer cells expressing FcRs are involved in the specific killing of target cells and / or the presentation of antigens to other components of the immune system, or their binding to antigen-presenting cells. In some embodiments, ADCC can be further enhanced by antibody-driven classical complement activation, which leads to the attachment of activated C3 fragments to target cells. C3 cleavage products are ligands for complement receptors (CRs), e.g., CR3, expressed on myeloid cells. Recognition of complement fragments by CRs on effector cells may promote the enhancement of Fc receptor-mediated ADCC. In some embodiments, antibody-driven classical complement activation leads to C3 fragments on target cells. These C3 cleavage products may promote direct complement-dependent cell-mediated cytotoxicity (CDCC). In some embodiments, effector cells can phagocytose target antigens, target particles, or target cells, which may be antibody-binding-dependent and mediated by FcγRs expressed by effector cells. The expression of specific FcRs or complement receptors on effector cells may be regulated by humoral factors such as cytokines. For example, FcγRI expression is known to be upregulated by interferon-gamma (IFNγ) and / or G-CSF. This enhanced expression increases the cytotoxicity of FcγRI-containing cells against the target. Effector cells can phagocytose the target antigen or phagocytose or lyse target cells.In some embodiments, antibody-driven classical complement activation introduces C3 fragments onto target cells. These C3 cleavage products can either directly promote phagocytosis by effector cells or indirectly enhance antibody-mediated phagocytosis.

[0075] As used herein, the term “Fc effector function” is intended to refer to a function resulting from the binding of a polypeptide or antibody to its target on the cell membrane, e.g., an antigen, where the Fc effector function is attributed to the Fc region of the polypeptide or antibody. Examples of Fc effector functions include: (i) C1q binding, (ii) complement activation, (iii) complement-dependent cell-mediated injury (CDC), (iv) antibody-dependent cell-mediated injury (ADCC), (v) Fcγ receptor binding, (vi) antibody-dependent cell phagocytosis (ADCP), (vii) complement-dependent cell-mediated injury (CDCC), (viii) complement-enhancing cell-mediated injury, (ix) antibody-mediated binding of opsonized antibodies to complement receptors, (x) opsonization, (xi) trogocytosis, and (xii) any combination of (i) to (xi).

[0076] As used herein, the term “complement activation” or “complement system activation” refers to the activation of the classical complement pathway initiated by a large macromolecular complex called C1 that binds to an antibody-antigen complex on a surface. C1 is a complex consisting of six recognition proteins C1q and a serine protease heterotetramer C1r2C1s2. C1 is the first protein complex in the initial events of the classical complement cascade, involving a series of cleavage reactions beginning with the cleavage of C4 into C4a and C4b, and C2 into C2a and C2b. C4b attaches and, together with C2a, forms an enzymatically active convertase called C3 convertase; C3 convertase cleaves complement component C3 into C3b and C3a to form C5 convertase. This C5 convertase splits C5 into C5a and C5b, with the last component adhering to the membrane; this then triggers a late complement activation event, in which terminal complement components C5b, C6, C7, C8, and C9 assemble to form the membrane attack complex (MAC). This complement cascade leads to pore formation in the cell membrane and causes cell lysis, also known as complement-dependent cell injury (CDC). Complement activation can be evaluated using the potency of C1q, the CDC kinetics assay (described in WO2013 / 004842, WO2014 / 108198), or by the method of cellular deposition of C3b and C4b described in Beurskens et al., J Immunol April 1, 2012 vol. 188 no. 7, 3532-3541.

[0077] Complement activation in a subject can be measured by determining, for example, the levels of C2 or CH50 according to any method known in the art. For example, C2 levels can be determined in plasma using a radioimmunodiffusion (RID) assay, and CH50 (complement lysis activity) can be measured in serum using a spectrophotometric assay with Autokit CH50.

[0078] As used herein, the term “complement-dependent cell injury” (CDC) is intended to refer to the process of antibody-mediated complement activation that results in the lysis of antibody-bound cells, a process that, while not theoretically bound, is considered to be a result of membrane pores formed by the assembly of so-called membrane invasion complexes (MACs). Appropriate assays for evaluating CDC are known in the art, including, for example, in vitro assays using normal human serum as a complement source, as described in Example 3. Non-limiting examples of assays for determining the maximum lysis, or EC50 value, of CD38-expressing cells mediated by a CD38 antibody may include the following steps: (a) Placing approximately 100,000 CD38-expressing cells per well in 40 μL of culture medium supplemented with 0.2% BSA in a multi-well plate; (b) Pre-incubating cells with 40 μL serially diluted CD38 antibody (0.0002–10 μg / mL) for 20 minutes; (c) Incubate each well with 20% pooled normal human serum at 37°C for 45 minutes; (d) Add a viability-determinating dye and measure the percentage of cell lysis using a flow cytometer; (e) A step in which the maximum dissolution is determined and / or the EC50 value is calculated using nonlinear regression.

[0079] As used herein, the term “antibody-dependent cell-mediated cytotoxicity” (“ADCC”) refers to the mechanism by which cells expressing an Fc receptor that recognizes the constant region of a bound antibody kill antibody-coated target cells. Suitable assays for evaluating ADCC are known in the Art, including, for example, the assay described in Example 4. Non-limiting examples of assays for determining ADCC in CD38-expressing cells mediated by a CD38 antibody are described below. 51 This may include a Cr release assay or a reporter assay step.

[0080] 51 ADCC by Cr release assay (a) In a multi-well plate, approximately 5,000 cells are placed in 50 μL of culture medium with 0.2% BSA added per well. 51 The step of plating Cr-labeled CD38-expressing cells (e.g., Daudi cells); (b) Pre-incubating cells for 15 minutes with 50 μL serially diluted CD38 antibody (0.0002-10 μg / mL); (c) Incubate each well with 500,000 newly isolated peripheral blood mononuclear cells (PBMCs) per well at 37°C for 4 hours; (d) Gamma counter of 75 μL of supernatant 51 A step in which the amount of Cr released is measured; (e) The step of calculating the cell lysis rate as (cpm sample - cpm spontaneous lysis) / (cpm maximum lysis - cpm spontaneous lysis), where cpm is the counts per minute.

[0081] ADCC by reporter assay (a) Plating approximately 5,000 CD38-expressing cells (e.g., Daudi cells) in 10 μL of standard medium (e.g., RPMI 1640) supplemented with 25% low-IgG serum in a multi-well plate suitable for optical reading (e.g., 384-well OptiPlates from PerkinElmer Inc.); (b) Incubate each well at 37°C for 6 hours with 10 μL of genetically modified Jurkat cells that stably express the NFAT response element driving the expression of FcγRIIIa receptor, V158 (high affinity) variant, and firefly luciferase as effector cells, and 10 μL of serially diluted CD38 antibody (0.0002-10 μg / mL); (c) Each well is incubated with 30 μL of luciferase substrate at room temperature for 5 minutes, and the luminescence is measured.

[0082] As used herein, the term “antibody-dependent cell phagocytosis” (“ADCP”) refers to a mechanism by which antibody-coated target cells are eliminated by internalization by phagocytic cells. Internalized antibody-coated target cells are contained within vesicles called phagosomes, which subsequently fuse with one or more lysosomes to form phagolysosomes. Appropriate assays for evaluating ADCP are known in the art, including, for example, in vitro cytotoxicity assays using macrophages as effector cells, video microscopy as described by van Bij et al. in the Journal of Hepatology Volume 53, Issue 4, October 2010, Pages 677-685, and the in vitro cytotoxicity assay described in Example 5. Non-limiting examples of assays for determining ADCP in CD38-expressing cells mediated by CD38 antibodies may include the following steps: (a) A step in which newly isolated monocytes are differentiated into macrophages by incubation in GM-CSF-containing medium for 5 days; (b) Placing approximately 100,000 macrophages per well in a multi-well plate onto GM-CSF-containing dendritic cell medium; (c) Adding 20,000 CD38 antibody-opsonized CD38-expressing cells (e.g., Daudi cells) labeled with a common fluorescent film dye per well at 37°C for 45 minutes; (d) A step in which the percentage of CD14-positive, CD19-negative, and membrane-dye-positive macrophages is measured using a flow cytometer.

[0083] As used herein, “trogocytosis” refers to a process characterized by the transfer of cell surface molecules from donor cells to acceptor cells, such as effector cells. Typical acceptor cells include T cells, B cells, monocytes / macrophages, dendritic cells, neutrophils, and NK cells. The trogocytosis-mediated transfer of cell surface molecules, such as CD38, from donor cells to acceptor cells can also result in the transfer of antibody-antigen complexes from donor cells to acceptor cells, i.e., the transfer of antibody-antigen complexes in which antibodies are bound to cell surface molecules. In particular, a special form of trogocytosis may occur when the acceptor cells are effector cells expressing the Fcγ receptor (FcγR); such acceptor cells can internalize donor cell-associated immune complexes, which typically consist of specific antibodies bound to a target antigen on the donor cell after FcγR has bound to the Fc region of the antibody. Appropriate assays for evaluating trogocytosis are known in the art, including, for example, the assay in Example 8. Non-limiting examples of assays for determining trogocytosis of CD38-expressing cells mediated by CD38 antibodies include:

[0084] Trogocytosis (Daudi cells): (a') The step of differentiating newly isolated monocytes into macrophages using GM-CSF for 5 days; (b') The step of plating approximately 100,000 macrophages per well into GM-CSF-containing dendritic cell medium; (c') Add approximately 20,000 CD38 antibody-opsonized Daudi cells labeled with a common fluorescent film dye to each well and incubate at 37°C for 45 minutes; (d') Steps to measure CD38 expression on Daudi cells using a flow cytometer, where a decrease in CD38 on CD38 antibody-opsonized Daudi cells compared to the control indicates trogocytosis.

[0085] Trogocytosis (Treg): (a) Placing approximately 500,000 newly isolated PBMCs per well into cell culture medium and leaving it overnight (O / N) at 37°C; (b) Adding approximately 100,000 CD38 antibody-opsonized Treg cells labeled with a common fluorescent intracellular amine dye per well and leaving them overnight (O / N) at 37°C; and (c) Steps to measure CD38 expression on Tregs using a flow cytometer, wherein a decrease in CD38 on CD38 antibody-opsonized Tregs compared to a control indicates trogocytosis.

[0086] A control may be selected by a person skilled in the art based on the specific purpose of the study or assay. Non-limiting examples of a control include (i) the absence of an antibody and (ii) an isotype control antibody. An example of an isotype control antibody is antibody b12, which has the VH and VL sequences listed in Table 4. In some embodiments where it is desired to evaluate the trogocytotic activity of the antibodies described herein, the control may be (iii) a parent antibody or reference antibody having different antigen-binding regions and / or different Fc regions.

[0087] In some embodiments, in step (b), the Tregs are labeled with a common fluorescent membrane dye in addition to, or instead of, an intracellular fluorescent amine dye.

[0088] In some embodiments, the reduction of CD38 antibody on donor cells can also be measured in steps (d') and (c) of the trogocytosis assay outlined above. For example, if the CD38 antibody is a human IgG (huIgG) antibody, huIgG can be detected using a secondary antibody.

[0089] In addition to Daudi cells (ATCC CCL-213), tumor cells suitable for the first assay include, but are not limited to, those listed in Table 2 of WO 2020 / 012036 A1 (incorporated herein by reference), particularly those with high CD38 expression.

[0090] In addition to Treg cells, CD38-expressing cells suitable for the second assay include, for example, immune cells such as NK cells, B cells, T cells, and monocytes, as well as tumor cells, particularly those with low CD38 expression levels, as listed in Table 2 of WO 2020 / 012036 A1.

[0091] As used herein, the term “vector” is intended to refer to a nucleic acid molecule capable of inducing the transcription of a nucleic acid segment ligated within the vector. One type of vector is a “plasmid,” which takes the form of a circular double-stranded DNA loop. Another type of vector is a viral vector, in which case the nucleic acid segment may be ligated to a viral genome. Certain types of vectors have the ability to autonomously replicate within the host cell into which they are introduced (e.g., bacterial vectors with bacterial origins of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) can be integrated into the host cell's genome upon introduction into the host cell and thereby replicate together with the host genome. Furthermore, certain types of vectors have the ability to induce the expression of a gene functionally ligated to them. Such vectors are referred to herein as “recombinant expression vectors” (or simply “expression vectors”). Generally, expression vectors useful in recombinant DNA technology are often in the form of plasmids. Hereinafter, “plasmid” and “vector” may be used interchangeably, as plasmids are the most commonly used form of vector. However, the present invention is intended to include other forms of expression vectors that perform equivalent functions, such as viral vectors (e.g., replication-deficient retroviruses, adenoviruses, and adeno-associated viruses).

[0092] As used herein, the term “recombinant host cell” (or simply “host cell”) is intended to refer to a cell into which one or more expression vectors have been introduced. For example, the HC and LC of the antibodies described herein may both be encoded by the same expression vector, and the host cell is transfected with that expression vector. Alternatively, the HC and LC of the antibodies described herein may be encoded by different expression vectors, and the host cell is cotransfected with those expression vectors. It should also be understood that the term “host cell” is intended to refer not only to a specific target cell but also to the offspring of such a cell. Because certain modifications may occur in subsequent generations due to mutation or environmental influences, such offspring may not actually be identical to the parent cell, but are nevertheless included within the scope of the term “host cell” as used herein. Recombinant host cells include, for example, transfectomas such as CHO cells, HEK-293 cells, PER.C6 cells, NS0 cells, lymphocytes, prokaryotic cells such as Escherichia coli (E. coli), and other eukaryotic cell hosts such as plant cells and fungi.

[0093] As used herein, the term "transfectoma" includes recombinant eukaryotic host cells expressing Ab or a target antigen, such as CHO cells, PER.C6 cells, NS0 cells, HEK-293 cells, plant cells, or fungi such as yeast cells.

[0094] The term "treatment" refers to administering an effective amount of the therapeutically active antibody of the present invention with the aim of alleviating, improving, preventing, or eradicating (curing) a symptom or condition.

[0095] The term "effective dose" or "therapeutic effective dose" refers to the effective amount of an antibody administered over the required dosage and duration to achieve the desired therapeutic outcome. The therapeutic effective dose of an antibody can vary depending on factors such as the individual's medical condition, age, sex, and weight, as well as the antibody's ability to elicit the desired response in the individual. The therapeutic effective dose is also the amount of an antibody whose therapeutically beneficial effects outweigh its toxicity or adverse effects.

[0096] As used herein, the terms “to bind” or “can bind” typically refer to the binding of an antibody to a given antigen or epitope, when determined by biolayer interferometry (BLI), or, for example, by surface plasmon resonance (SPR) technique using a BIAcore 3000 instrument with the antigen as the ligand and the antibody as the analyte, approximately 10 -7 M or less, for example, about 10 -8 M or less, for example, about 10 -9 M or less, about 10 -10 M or less, or about 10 -11 M or even lower K D This is binding at an affinity corresponding to the antibody. The antibody binds to nonspecific antigens other than the given antigen or closely related antigens (e.g., BSA, casein) at an affinity corresponding to the antibody's K D K that is at least 10 times lower, for example, at least 100 times lower, for example, at least 1,000 times lower, for example, at least 10,000 times lower, for example, at least 100,000 times lower D It binds to a predetermined antigen with an affinity corresponding to the K of binding. D The amount that becomes lower is the K of the antibody D Because it depends on the K of the antibody D If the K of antigen binding is very low, D However, the K of binding to nonspecific antigens D The reduction in specificity can be at least 10,000 times (i.e., the antibody is highly specific).

[0097] The term "k" as used herein d (sec 1 ) refers to the dissociation rate constant of a specific antibody-antigen interaction. The value is k off Also called a value.

[0098] The term "K" as used in this specification D (M) refers to the dissociation equilibrium constant of a specific antibody-antigen interaction. Affinity and K as used herein D They are inversely correlated, meaning that an increase in affinity is K D This is intended to refer to a decrease in K DThis is intended to refer to an increase in [a certain value].

[0099] In this specification, a "treatment cycle" is defined as the period within which the effects of individual antibody administrations are exerted by their pharmacodynamic action, or in other words, the period during which the administered antibody is essentially eliminated from or in the process of being eliminated from the subject's body. Short timeframes; for example, within 2 to 24 hours, for example, within 2 to 12 hours, or multiple small doses on the same day may be equivalent to more single doses.

[0100] The efficacy of the treatment for multiple myeloma (MM) can be evaluated according to the unified efficacy criteria for response and minimal residual disease assessment in multiple myeloma (Kumar et al., 2016) established by the International Myeloma Working Group (IMWG) in 2016, as shown in Table 1 below.

[0101] (Table 1) Unified response criteria for evaluating response and minimal residual disease in multiple myeloma, as established by the International Myeloma Working Group (IMWG) in 2016 (Kumar et al., 2016) TIFF2026513656000001.tif166167TIFF2026513656000002.tif186167TIFF2026513656000003.tif193167CR = Complete Response;CRAB = Elevated Calcium, Renal Failure, Anemia, and Bone Lesions;DFS = Disease-Free Survival Period;CT = Computed Tomography;FCM = Flow Cytometry;FDG = Fluorodeoxyglucose;FLC = Free Light Chain;h = Time;IFE = Immunofixation;IHC = Immunohistochemical Testing mAb = monoclonal antibody; MM = multiple myeloma; MR = minimal response; MRD = minimal residual disease; NGF = next-generation flow sequencing; NGS = next-generation sequencing; MFC = multi-parameter flow cytometry; PD = progression; PET = positron emission tomography; PFS = progression-free survival; PR = partial response; sCR = severe complete response; SD = stable; SPD = sum of the products of the largest orthogonal diameters of the lesions measured; SUV = standardized uptake; SUVmax = maximum standardized uptake; VGPR = best partial response

[0102] 1. All efficacy categories require two consecutive evaluations performed at any point before the initiation of any new treatment; for MRD, two consecutive evaluations are not required, but information on MRD after each treatment phase is recommended (e.g., after induction, high-dose treatment / ASCT, consolidation, maintenance); MRD testing should only be initiated when CR is suspected. All categories of response and MRD require no known evidence of progression or new bone lesions if radiographic imaging is performed. However, if radiographic imaging reports a negative MRD status, radiographic imaging is not required to meet these requirements for FDG PET. 2. If persistent MRD negativity is reported, the method used should also be noted (e.g., persistent flow MRD negativity, persistent sequence MRD negativity). 3. Bone marrow MFCs should follow NGF guidelines. The reference NGF method is an 8-color, 2-tube approach and is widely validated. The 2-tube approach improves reliability, consistency, and sensitivity by obtaining a larger number of cells. 8-color technology is widely available worldwide, and the NGF method is already adopted by many flora laboratories around the world. The complete 8-color method is the most efficient method using lyophilized antibody mixtures, reducing errors, time, and costs. 5 million cells should be evaluated. The FCM method to be used is 10 5 It should have a detection sensitivity of at least one per plasma cell. 4. DNA sequencing assays of bone marrow aspirates should use validated assays such as LymphoSIGHT (Sequenta). 5. Criteria used by Zamagni et al. and the expert committee (IMPetUs; Italian Myeloma Criteria for PET Use). Baseline-positive lesions were identified by the presence of an increased uptake area within the bone, regardless of the presence of any underlying lesions identified by CT and present on at least two consecutive sections. Alternatively, SUVmax=2.5 was considered positive within the range of osteolytic CT area >1 cm size, or SUVmax=1.5 within the range of osteolytic CT area ≤1 cm size. If MRD is determined to be negative by MFC or NGS, imaging studies should be performed. 6. Based on internationally unified criteria for evaluating response. The definitions and classifications of lower-level responses are based on Rajkumar et al. When the only method for measuring the disease is serum FLC levels: CR can be defined as a normal FLC ratio of 0.26 to 1.65, in addition to the aforementioned CR criteria. VGPR in such patients requires a reduction of ≥90% in the difference between involved and uninvolved FLC levels. All response categories require two consecutive evaluations performed at any point before the initiation of any new treatment; all categories also require no known evidence of progression or new bone lesions or extramedullary plasmacytoma, if radiographic imaging is performed. Radiographic imaging is not required to meet these response requirements. Confirmation of bone marrow evaluation is not required. Each category except SD is considered unconfirmed until a definitive test is performed. For time-dependent outcome evaluations such as duration of response, the date of the first test is considered the response date. 7. All recommendations regarding clinical use related to serum FLC levels and FLC ratios are based on the results obtained from the validated Freelight trial (Binding Site, Birmingham, UK). 8. The presence or absence of clonal cells in IHC is based on the κ / λ / L ratio. An abnormal κ / λ ratio detected by IHC requires a minimum of 100 plasma cells for analysis. Abnormal ratios reflecting the presence of abnormal clones are κ / λ >4:1 or <1:2. 9. Particular attention should be paid to the appearance of different monoclonal proteins post-treatment, often associated with oligoclonal rearrangement of the immune system, especially in patients who have achieved conventional complete response (CR). These bands typically disappear over time and have been associated with better outcomes in some studies. Furthermore, the appearance of monoclonal IgGκ in patients treated with mAbs should be distinguished from that of therapeutic antibodies. 10. Measurements of plasmacytoma should be obtained from the CT portion of PET / CT, or from an MRI scan, or a dedicated CT scan if available. In patients with only skin lesions, the skin lesions should be measured with a ruler. Tumor size measurements are determined by SPD. 11. In patients previously classified as having achieved CR, if only IFE is positive, it is not considered progression. For the purpose of calculating progression-free survival and PFS, patients who have achieved CR and are MRD-negative should be evaluated using the criteria listed under PD. The criteria for relapse from CR or relapse from MRD should only be used when calculating disease-free survival. 12. If a value is perceived by a physician as an erroneous result (e.g., a possible test error), that value will not be considered when determining the minimum value.

[0103] As used herein, “Best Overall Response” (BOR) refers to the best effect recorded during the course of treatment in a clinical trial. Subjects with sCR, CR, VGPR, or PR are considered to have an objective response.

[0104] As used herein, "objective response rate (ORR)" refers to the percentage of subjects who achieve a partial response or better (e.g., PR, VGPR, CR, or sCR).

[0105] As used herein, the "clinical benefit rate" (CBR) is defined as the proportion of subjects who have at least stability.

[0106] As used herein, “Duration of Response (DOR)” applies only to subjects with a confirmed best overall response of PR or better (e.g., PR, VGPR, CR, or sCR), and is defined as the time from the first record of objective tumor response (e.g., PR, VGPR, CR, or sCR) to the date of MR, SD, PD, or death from the underlying cancer.

[0107] As used herein, "Time to Response (TTR)" is defined as the time from C1D1 to the occurrence of a response (PR or better). The TTR is descriptively summarized and presented for subjects with a response (PR or better).

[0108] Progression-free survival (PFS) is defined as the time, e.g., days, from day 1 of cycle 1 (C1D1) to the first recorded death due to progression or any other cause.

[0109] As used herein, "overall survival (OS)" is defined as the time, e.g., in days, from day 1 of cycle 1 (C1D1) to death from any cause. If it is not known that a subject has died, OS is terminated on the latest day (before the cutoff date) on which it is known that the subject was alive.

[0110] As used herein, “adverse event (AE)” refers to any undesirable medical event in a patient or clinical trial subject that is time-related to the use of a drug, whether or not it is related to the drug. Therefore, an AE may be any undesirable and unintended sign (including abnormal clinical laboratory values), symptom, or disease (new or worsening) that is time-related to the use of a drug. The severity of AEs is described according to the Common Terminology Criteria for Adverse Events (CTCAE), v5.0 (available at https: / / ctep.cancer.gov / protocolDevelopment / electronic_applications / docs / CTCAE_v5_Quick_Reference_8.5x11.pdf), with the exception of TLS (tumor lysis syndrome), which is classified according to Cairo-Bishop et al (Coiffier, B., Altman, A., Pui, CH, Younes, A., and Cairo, MS (2008). Guidelines for the management of pediatric and adult tumor lysis syndrome: an evidence-based review. J Clin Oncol 26, 2767-2778). AEs include only TEAEs, i.e., AEs that began during the treatment period or pre-existing AEs that worsened during the treatment period.

[0111] As used herein, AEs include "Serious Adverse Events (SAEs)" which are defined as AEs that meet at least one of the following criteria: • Anything that is deadly or life-threatening • Those that result in permanent or significant impairment / dysfunction. • Constituent elements of congenital anomalies / congenital defects • Defined as an event that is medically significant, i.e., one that endangers the subject or may require medical or surgical intervention to prevent one of the above outcomes. • Those requiring hospitalization or extension of existing hospitalization.

[0112] In the definition of SAE, the term "life-threatening" refers to an event in which there was a risk of death for the subject at the time of the event; it does not refer to an event that, if more severe, could hypothetically have resulted in death.

[0113] Furthermore, according to the definition of SAE, hospitalization for the following reasons should not be reported as an SAE: • Routine treatment or monitoring of the underlying condition • Caused solely by the progression of the underlying cancer. • Elective or pre-planned treatment for pre-existing conditions that are unrelated to the underlying disease and have not worsened since the signing of informed consent. • No deterioration in the subject's overall condition, social reasons, and respite care. Treatment in the emergency room for events that do not require hospitalization and do not fall under any of the above definitions of SAE is not considered SAE.

[0114] AEs also include “Adverse Events of Special Interest (AESIs),” which are defined as events (serious or non-serious) that are scientific and medical concerns specific to the sponsor’s product or program and may warrant continued monitoring and prompt communication by the principal investigator. Such events may require further investigation to characterize and understand them. Infusion reactions (IRRs) are considered AESIs. Other AESIs are defined based on the ongoing review of safety data.

[0115] As used herein, “infusion reaction” (IRR) is defined as any AE that occurs during infusion or begins within 24 hours after the completion of infusion. In an IRR, the causal relationship of the event should be determined to be “related” by the principal investigator.

[0116] As used herein, "tumor lysis syndrome" (TLS) is characterized by a series of metabolic abnormalities resulting from the rapid lysis of malignant cells followed by the massive and rapid release of cellular components into the bloodstream, and is defined according to the Cairo-Bishop classification (Coiffier et al., 2008, above).

[0117] As used herein, “dose-limiting toxicity” (DLT) refers to any adverse event of a specified grade, except those that are clearly and unquestionably due to an underlying disease or external cause. In this context, the DLT evaluation period is defined as the first 28 days of treatment (i.e., cycle 1). Toxicity, with the exception of TLS, is classified for severity according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI-CTCAE), v5.0. A subject is said to be “DLT evaluable” after meeting the minimum exposure criteria (i.e., receiving all four pre-planned doses during the DLT evaluation period; note that the first two divided doses on days 1 and 2 are equivalent to one pre-planned dose) and having a sufficient safety assessment (i.e., completing the DLT evaluation period) or after experiencing DLT during the first 28 days of administration (i.e., cycle 1). A subject that does not meet the “DLT evaluable” criteria is considered “DLT unevaluable.”

[0118] As used herein, “Maximum Tolerable Dose” (MTD) refers to the highest dose level (DL) that produces a tolerable level of toxicity (i.e., within the target toxicity range) based on the mBOIN algorithm.

[0119] As used herein, “previously untreated” refers to subjects who have never received prior anti-cancer treatment (for certain hematological malignancies), i.e., are naive to a particular treatment. For example, a subject who has never been treated with any anti-CD38 antibody is anti-CD38 naive, a subject who has never been treated with daratumumab is daratumumab naive ("dara naive"), and a subject who has never been treated with isatuximab is isatuximab naive ("isa naive"). A subject who is anti-CD38 naive may or may not have received prior anti-cancer treatment with a drug other than an anti-CD38 antibody, but has never received prior treatment with an anti-CD38 antibody. A subject who is dara naive may or may not have received prior anti-cancer treatment, but has never received prior treatment with an anti-CD38 antibody. Similarly, a subject who is isa naive may or may not have received prior anti-cancer treatment, but has never received prior treatment with an anti-CD38 antibody.

[0120] In the context of this invention, the term "treatment regimen" refers to a systematic treatment plan designed to improve and maintain health.

[0121] The terms "pharmaceutical composition" and "pharmaceutical preparation" are used interchangeably in this specification.

[0122] "Approximately" means that a particular value is within the tolerance range determined by those skilled in the art, and this depends in part on the method of measuring or determining the value, i.e., the limits of the measuring system. In the context of a particular assay, result, or embodiment, unless otherwise specified within the scope of the examples or elsewhere in the specification, "approximately" means a range of up to 5%.

[0123] Specific aspects of the present invention Various aspects of the present invention are described below herein. The present invention is based on the observation that an anti-CD38 antibody containing a mutation in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 in the human IgG1 heavy chain, for example E430G, and particularly the antibody IgG1-C-E430G, is safe and well-tolerated when used in humans, particularly in the treatment of hematological malignancies such as multiple myeloma (MM), and exhibits biological activity and efficacy at a dose level of 4 mg / kg body weight.

[0124] As established during dose escalation, no dose-level toxicity (DLT) was observed at dose levels ranging from 0.2 / 0.6 mg / kg to 16 mg / kg. Tumor lysis syndrome (TLS) or cytokine release syndrome (CRS) was not observed up to a dose level of 24 mg / kg. The most commonly observed adverse events (AEs) under the study treatment were infusion reactions (IRR; 75.0%), neutropenia (62.5%), anemia (41.7%), diarrhea (41.7%), COVID-19 (25.0%), fever (25.0%), thrombocytopenia (20.8%), and blurred vision (20.8%). Most IRRs were low-grade (G; G1-2: 58.3%, G3: 16.7%, G4: none), occurred mainly at the initial infusion, and were manageable.

[0125] In 19 patients (n=5, naive to anti-CD38 mAb; n=16, refractory to anti-CD38 mAb) who were evaluable with dose escalation, preliminary antitumor activity was observed. In patients naive to anti-CD38 mAb, two patients achieved complete response (CR; one each at 4 mg / kg and 24 mg / kg), and one achieved minimal response (MR; 16 mg / kg). In patients refractory to anti-CD38 mAb, one patient achieved partial response (PR) (16 mg / kg), and two patients achieved MR (one each at 8 mg / kg and 16 mg / kg).

[0126] The biological activity of the antibody was confirmed at all dose levels evaluated during dose escalation. A rapid and sustained decrease in peripheral blood natural killer cells was observed at all doses in all patients (median peak decrease from baseline [BL] 97%; range 66–100%, n=21, patients who only received BL evaluation were excluded from analysis). T cells transiently decreased after administration of the initial dose ≥4 mg / kg, particularly in patients naive to anti-CD38 mAb, and then expanded and proliferated (≥100% increase from BL). Transient reduction of complement component C2 (median peak reduction from BL 64%; range 6–78%, n=18; patients with BL values ​​below LLOQ; patients with no BL value or only BL value were excluded from this analysis) and total complement lysis activity (median peak reduction from BL 53% at ≥8 mg / kg; range 2–92%, n=11; patients with BL values ​​below LLOQ; patients with no BL value or only BL value were excluded from this analysis) were induced at all evaluable doses, suggesting complement-mediated response (CDC). Complement parameters rapidly recovered to BL levels, indicating that the treatment did not deplete complement. Plasma cytokine (IL-2, IL-6, IL-8, IL-10, IFNγ, and TNFα) levels generally remained low post-treatment. In particular, complement activation appeared to be enhanced compared to previous CD38 antibodies at similar dose levels (Nijhof et al., Blood 2016 Aug 18;128(7):959-70).

[0127] Surprisingly, no dose-dependent decrease was observed in CD38-expressing non-tumor cells (other than NK cells), such as B cells, T cells, monocytes, and / or NKT-like cells. PK data from dose escalation in 22 subjects administered doses of 0.2–24 mg / kg showed that the PK level was lower at the maximum concentration (C max This indicates that the AUC (AUC) increased in approximately proportion to the dose. 0-tThe drug concentration (DL) increased more than proportionally at doses up to 4 mg / kg, but increased roughly proportionally at higher dose levels. No increase in mean AUC was observed at 16 mg / kg–24 mg / kg. These observations suggest clear target-mediated drug elimination at DL levels below 4 mg / kg, and a high degree of target saturation over a one-week administration period at approximately 4 mg / kg and above. At 16 mg / kg, the PK profile was more consistent across subjects, and exposure was better maintained with bi-weekly administration compared to lower dose levels. Accumulation of peak concentrations after the initial full dose was limited in all DL levels, suggesting relatively rapid linear clearance. Signs of increased trough concentrations between weekly administrations were observed in the PK profile, indicating reduced clearance, possibly due to a decrease in CD38 due to target cell depletion.

[0128] In particular, the clearance of this antibody at doses of 4–24 mg / kg in the DL was observed to be faster than that observed with the previous CD38 antibodies daratumumab (Clemens et al., Immunomodulatory Drug Treatment. Clin Pharmacokinet. 2017 Aug;56(8):915-924) and isatuximab (Martin et al., Blood Cancer J. 2019 Mar 29;9(4):41) in similar dose ranges. This was especially surprising, as the clearance of other IgGs with similar hexamerization-promoting mutations had previously been observed to be similar to that of normal IgG in mice (De Jong et al., PLoS Biol. 2016 Jan 6;14(1):e1002344).

[0129] These results were confirmed in expanded Part A of the clinical trial, where subjects with RRMM who were anti-CD38 mAb naive were treated with RP2D (16 mg / kg), which was identified for RRMM in the dose-escalation part of the clinical trial.

[0130] The best overall response among the 11 subjects was complete response in one subject (9.1%), best partial response in two subjects (18.2%), partial response in three subjects (27.3%), minimal response in two subjects (18.2%), and stable response in one subject (9.1%). Two subjects could not be evaluated.

[0131] The most common adverse events (TEAEs) observed in the expanded phase were neutropenia (6 subjects; 54.5%), as well as anemia, headache, IRR, thrombocytopenia, and upper respiratory tract infections (3 subjects per event; 27.3%).

[0132] IRR (27.3%) was manageable at Grade 2, and treatment was not discontinued. No events of cytokine release syndrome were reported.

[0133] Treatment was accompanied by a transient reduction in complement component C2 and overall complement-mediated lysis activity in the majority of patients, suggesting CDC activity. Transient T cell decrease and T cell proliferation (≥50% increase from baseline over ≥2 visits) were observed in 4 out of 10 evaluable patients after the initial dose in all patients.

[0134] Furthermore, PK data from the expansion phase were confirmed to be those observed during dose escalation. The PK profiles at 16 mg / kg were similar between the two cohorts. Peak and pre-administration concentrations were comparable between the cohorts.

[0135] Therefore, when the antibodies of the present invention are used in humans, for example, in the treatment of hematological malignancies such as MM, they surprisingly exhibit low, manageable side effects while simultaneously showing signs of enhanced antitumor activity (e.g., response and complement activation).

[0136] Dosage and treatment regimen As described above, the present invention relates to an anti-CD38 antibody, in particular an anti-CD38 antibody comprising an Fc region containing a mutation in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 in the human IgG monohepatic chain. In particular, such use of such an antibody for the treatment of one or more hematological malignancies (e.g., MM) in a subject is described herein.

[0137] Therefore, in the first aspect, the present invention provides a method for treating or preventing hematological malignancies, preferably multiple myeloma (MM), in subjects in need, preferably human subjects, comprising the step of administering an antibody or a pharmaceutical composition containing the antibody to the subject in a therapeutically effective amount. The antibody, a. An antigen-binding region comprising VH CDR1 having the sequence described in SEQ ID NO:2, VH CDR2 having the sequence described in SEQ ID NO:3, VH CDR3 having the sequence described in SEQ ID NO:4, VL CDR1 having the sequence described in SEQ ID NO:6, VL CDR2 having sequence AAS, and VL CDR3 having the sequence described in SEQ ID NO:7, and a. Fc region containing mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 (amino acid residues are numbered according to the EU index) in the human IgG monohelic acid. This provides a method that includes [something].

[0138] In another aspect, the present invention provides a method for treating or preventing hematological malignancies, preferably multiple myeloma (MM), in subjects in need, preferably human subjects, comprising the step of administering an antibody or a pharmaceutical composition containing the antibody to the subject in a therapeutically effective amount. The antibody, a. A heavy chain comprising a VH region containing VH CDR1 having the sequence described in SEQ ID NO:2, VH CDR2 having the sequence described in SEQ ID NO:3, and VH CDR3 having the sequence described in SEQ ID NO:4, and a human IgG1 CH region having mutations in one or more of E430, E345, and S440 (amino acid residues are numbered according to the EU index); and b. A light chain comprising a VL region including VL CDR1 having the sequence described in SEQ ID NO:6, VL CDR2 having sequence AAS, and VL CDR3 having the sequence described in SEQ ID NO:7. This provides a method that includes [something].

[0139] In a preferred embodiment, the amount of antibody administered (at each dose and / or at each treatment cycle) is at least (approximately) 4 mg / kg body weight, for example, the antibody is administered in doses of at least (approximately) 4 mg / kg body weight to (approximately) 24 mg / kg body weight.

[0140] In one embodiment, the dosage is in the range of approximately 4 mg / kg to approximately 24 mg / kg body weight.

[0141] In one embodiment, the dose is in the range of approximately 4 mg / kg to approximately 20 mg / kg body weight.

[0142] In one embodiment, the dosage is in the range of approximately 4 mg / kg to approximately 16 mg / kg body weight.

[0143] In one embodiment, the dose is in the range of approximately 4 mg / kg to approximately 12 mg / kg body weight.

[0144] In one embodiment, the dose is in the range of approximately 4 mg / kg to approximately 8 mg / kg body weight.

[0145] In one embodiment, the dosage is in the range of approximately 8 mg / kg to approximately 24 mg / kg body weight.

[0146] In one embodiment, the dose is in the range of approximately 8 mg / kg to approximately 20 mg / kg body weight.

[0147] In one embodiment, the dose is in the range of approximately 8 mg / kg to approximately 16 mg / kg body weight.

[0148] In one embodiment, the dose is in the range of approximately 12 mg / kg to approximately 24 mg / kg body weight.

[0149] In one embodiment, the dose is in the range of approximately 12 mg / kg to approximately 20 mg / kg body weight.

[0150] In one embodiment, the dose is in the range of approximately 12 mg / kg to approximately 16 mg / kg body weight.

[0151] In one embodiment, the dose is approximately 4 mg / kg body weight.

[0152] In one embodiment, the dose is approximately 6 mg / kg body weight.

[0153] In one embodiment, the dose is approximately 8 mg / kg body weight.

[0154] In one embodiment, the dose is approximately 10 mg / kg body weight.

[0155] In one embodiment, the dose is approximately 12 mg / kg body weight.

[0156] In one embodiment, the dose is approximately 14 mg / kg body weight.

[0157] In one embodiment, the dose is approximately 16 mg / kg body weight.

[0158] In one embodiment, the dose is approximately 18 mg / kg body weight.

[0159] In one embodiment, the dose is approximately 20 mg / kg body weight.

[0160] In one embodiment, the dose is approximately 22 mg / kg body weight. In another embodiment, the dose is approximately 24 mg / kg body weight.

[0161] In one embodiment, the dose is in the range of 4 mg / kg to 24 mg / kg body weight.

[0162] In one embodiment, the dose is in the range of 4 mg / kg to 20 mg / kg body weight.

[0163] In one embodiment, the dose is in the range of 4 mg / kg to 16 mg / kg body weight.

[0164] In one embodiment, the dose is in the range of 4 mg / kg to 12 mg / kg body weight.

[0165] In one embodiment, the dose is in the range of 4 mg / kg to 8 mg / kg body weight.

[0166] In one embodiment, the dose is in the range of 8 mg / kg to 24 mg / kg body weight.

[0167] In one embodiment, the dose is in the range of 8 mg / kg to 20 mg / kg body weight.

[0168] In one embodiment, the dose is in the range of 8 mg / kg to 16 mg / kg body weight.

[0169] In one embodiment, the dose is in the range of 12 mg / kg to 24 mg / kg body weight.

[0170] In one embodiment, the dose is in the range of 12 mg / kg to 20 mg / kg body weight.

[0171] In one embodiment, the dose is in the range of 12 mg / kg to 16 mg / kg body weight.

[0172] In one embodiment, the dose is 4 mg / kg body weight.

[0173] In one embodiment, the dose is 6 mg / kg body weight.

[0174] In one embodiment, the dose is 8 mg / kg body weight.

[0175] In one embodiment, the dose is 10 mg / kg body weight.

[0176] In one embodiment, the dose is 12 mg / kg body weight.

[0177] In one embodiment, the dose is 14 mg / kg body weight.

[0178] In one embodiment, the dose is 16 mg / kg body weight.

[0179] In one embodiment, the dose is 18 mg / kg body weight.

[0180] In one embodiment, the dose is 20 mg / kg body weight.

[0181] In one embodiment, the dose is 22 mg / kg body weight.

[0182] In one embodiment, the dose is 24 mg / kg body weight.

[0183] At higher DL (≥8 mg / kg), the PK profile (exposure) was observed to be more consistent among subjects. Therefore, in one embodiment, the antibody is administered at doses of approximately 8 mg / kg body weight to approximately 24 mg / kg body weight, for example, 8–24 mg / kg body weight.

[0184] Furthermore, at 16-24 mg / kg, exposure (AUC 0-t No increase in ) was observed. Therefore, in one embodiment, the antibody is administered in doses of approximately 8 mg / kg body weight to approximately 16 mg / kg body weight, for example, 8 to 16 mg / kg body weight. For example, the antibody is administered in doses of (approximately) 8 mg / kg body weight, or (approximately) 10 mg / kg body weight, or (approximately) 12 mg / kg body weight, or (approximately) 14 mg / kg body weight, or (approximately) 16 mg / kg body weight.

[0185] In a preferred embodiment, the antibody is administered at a dose of approximately 16 mg / kg body weight or 16 mg / kg body weight. This dose was found to be remarkably optimal compared to typical IgG1 antibodies or prior art CD38 antibodies, given, on the one hand, the relatively high binding affinity and CDC activity observed in vitro and the complement activation observed in vivo, and on the other hand, the unexpectedly high clearance of the antibody across all dose levels. Simultaneously, no tumor lysis syndrome (TLS) or cytokine release syndrome (CRS) was observed at any dose level, including 16 mg / kg, and adverse events (EA) were manageable. Furthermore, exposure was found to be better maintained at 16 mg / kg compared to 8 mg / kg, especially when the dosing interval was shortened, for example, every other week.

[0186] In any embodiment described herein, the dose, defined in mg / kg body weight, is preferably based on the subject's body weight measured at the time of administration of each dose, preferably within 72 hours prior to administration.

[0187] According to any of these embodiments, the dose defined in mg / kg may be converted to a fixed dose based on the median body weight of the subject to whom the binder is administered being 70 kg, and vice versa. Thus, the antibody may be administered in doses of approximately 250 to 2000 mg, for example, approximately 280 to 1700 mg. Therefore, any of the above doses per kg of body weight can thus be converted to a uniform dose.

[0188] Each treatment cycle is preferably about 4 weeks, or about 28 days.

[0189] In one embodiment, the dose is administered weekly (Q1W), preferably at least once, for example, two, three, four, five, six, seven, and more preferably eight times. Such weekly doses correspond to four doses in each cycle of about four weeks / about 28 days. Therefore, preferably, the weekly administration is carried out over two cycles of about four weeks / about 28 days.

[0190] In one embodiment, the weekly administration, or administration interval, may be shortened to one administration every two weeks (Q2W), also known as bi-weekly administration. Such bi-weekly administration may be performed at least once, for example, two, three, four, five, six, seven, and more preferably eight times. Such bi-weekly doses correspond to two doses in each cycle of approximately 4 weeks / approximately 28 days. Therefore, preferably, the bi-weekly administration is performed over four cycles of approximately 4 weeks / approximately 28 days, i.e., eight times (after the weekly administration).

[0191] After the aforementioned bi-weekly administration, the interval may be further shortened to once every four weeks (Q4W), i.e., once every (approximately) 28 days. Such four-weekly administrations may be carried out over a long period, preferably at least one, two, three, four, five, six, seven, eight, nine, ten, or more times. Such four-weekly doses correspond to one dose in each (approximately) four-week / (approximately) 28-day cycle. Therefore, preferably, the four-weekly administrations are carried out over a period of at least one, two, three, four, five, six, or more cycles, for example, eight, ten, twelve, fourteen, sixteen, eighteen, twenty, twenty-four, or more cycles (after the aforementioned weekly administrations, preferably after the aforementioned bi-weekly administrations).

[0192] In one embodiment, the initial dose (or starting dose) of the antibody is administered as a divided dose over two days, preferably over two consecutive days, meaning that one portion of the initial dose is administered on one day and the remainder on another day, preferably on day 1 (C1D1) and day 2 (C1D2) of the first cycle. Preferably, the divided doses are (approximately) equal. For example, if the dose is (approximately) 4 mg / kg body weight, the initial dose is administered to the subject as (approximately) 2 mg / kg on day 1 (C1D1) of cycle 1 and as (approximately) 2 mg / kg on the immediately following day, preferably the next day (i.e., day 2 of cycle 1: C1D2). Similarly, if the dose is (approximately) 8 mg / kg body weight, the initial dose is administered to the subject as (approximately) 4 mg / kg on day 1 (C1D1) of cycle 1 and as (approximately) 4 mg / kg on the immediately following day, preferably the next day (C1D2). Similarly, if the dose is approximately 16 mg / kg body weight, the initial dose is administered to the subject at approximately 8 mg / kg on day 1 of cycle 1 (C1D1) and then at approximately 8 mg / kg on the following day, preferably the next day (C1D2). Similarly, if the dose is approximately 24 mg / kg body weight, the initial dose is administered to the subject at approximately 12 mg / kg on day 1 of cycle 1 (C1D1) and then at approximately 12 mg / kg on the following day, preferably the next day (C1D2). Therefore, preferably, there is approximately 24 hours between the first portion of the initial (divided) dose and the second portion of the initial (divided) dose.

[0193] In a preferred embodiment, the antibody is administered in a cycle of approximately 28 days, or approximately 4 weeks, where the antibody is administered weekly (Q1W) in cycles 1 and 2, every 2 weeks (Q2W) in cycles 3-6, and every 4 weeks (Q4W) from cycle 7 onward, preferably the initial dose is administered as (equal) divided doses over the first two days.

[0194] In one embodiment, the antibody is administered for a sufficient amount of time to treat the hematological malignancy.

[0195] In another embodiment, antibodies are administered until the disease progresses or the patient no longer receives any benefit.

[0196] In one embodiment, the antibody is administered to the subject by intravenous injection or infusion.

[0197] In one embodiment, the antibody is administered by intravenous injection or infusion in a volume of approximately 100 ml to approximately 500 ml per dose, for example, approximately 100 ml or approximately 500 ml per dose.

[0198] In one embodiment, the antibody is administered by intravenous injection or infusion over a period of approximately 1 to 11 hours, or approximately 1 to 10 hours, or approximately 3 to 10 hours, for example, approximately 1 to 8 hours, or approximately 1 to 5 hours, for example, approximately 4 hours, per dose.

[0199] Disease and previous treatments The antibody of the present invention, or a pharmaceutical composition containing the antibody, may be used for the treatment or prevention of hematological malignancies or diseases or disorders. As used herein, “hematological malignancies” refers to cancers that originate in hematopoietic tissue, such as bone marrow, or in cells of the immune system. Examples of hematological cancers include leukemia, lymphoma, and multiple myeloma. Hematological malignancies are often also called blood cancers.

[0200] In one embodiment, the disease or disorder may be any hematological disorder or disorder described herein that involves cells expressing CD38. In other words, hematological malignancies may be CD38-positive hematological malignancies, i.e., hematological malignancies including leukemia, lymphoma and myeloma, or hematological malignancies known to express CD38, characterized by the presence of at least a portion of tumor cells expressing CD38. Examples of such CD38-positive hematological malignancies or hematological malignancies known to express CD38 include progenitor B-cell lymphoblastic leukemia / lymphoma and B-cell non-Hodgkin lymphoma, acute promyelocytic leukemia, acute lymphoblastic leukemia, and mature B-cell neoplasms, such as B-cell chronic lymphocytic leukemia (CLL) / small lymphocytic lymphoma (SLL), B-cell acute lymphoblastic leukemia, B-cell prolymphoblastic leukemia, lymphoplasmacytic lymphoma, mantle cell lymphoma (MCL), and follicular lymphoma. These include follicular lymphomas (FL), such as low-grade, intermediate-grade, and high-grade FL, cutaneous follicular lymphoma, marginal zone B-cell lymphoma (MALT type, lymph node and splenic type), hairy cell leukemia, diffuse large B-cell lymphoma (DLBCL), Burkitt lymphoma (BL), plasmacytoma, multiple myeloma, plasmacytoma, post-transplant lymphoproliferative disorders, light chain amyloidosis, Waldenström macroglobulinemia, plasmacytoma, and anaplastic large cell lymphoma (ALCL).

[0201] The antibodies of the present invention may be used for the treatment or prevention of hematological malignancies or disorders in subjects who have previously received at least one prior treatment for the same disease or disorder with one or more compounds different from the antibodies of the present invention.

[0202] For example, in some embodiments, the antibodies of the present invention may be for use in the treatment or prevention of a disease or disorder in subjects who have previously received treatment with proteasome inhibitors (PIs) and / or immunomodulatory agents (IMiDs). Examples of proteasome inhibitors include, but are not limited to, bortezomib, carfilzomib, and ixazomib. Examples of IMiDs include, but are not limited to, thalidomide, lenalidomide, and pomalidomide. In further embodiments, the disease or disorder may be cancer or tumor, for example, multiple myeloma, mantle cell lymphoma, or myelodysplastic syndrome (MDS). Therefore, the subjects may be cancer patients, for example, patients with multiple myeloma, mantle cell lymphoma, or myelodysplastic syndrome (MDS).

[0203] The antibodies of the present invention may be for use in the treatment or prevention of disease or impairment in subjects who have not received any prior treatment with anti-CD38 antibodies. Typically, such subjects or patients are referred to as anti-CD38 antibody-naive patients. In one embodiment, the anti-CD38 antibody is daratumumab and / or isatuximab; that is, the subject or patient has not received any prior treatment with daratumumab and / or isatuximab. Thus, in one embodiment, the subject or patient is a daratumumab-naive subject / patient or an isatuximab-naive subject / patient.

[0204] The present invention also provides antibodies according to the present invention for use in the treatment or prevention of hematological disorders in subjects who have received at least one prior treatment comprising a CD38 antibody. Such prior treatments may be one or more cycles of a planned treatment program comprising a CD38 antibody, e.g., one or more planned cycles of a CD38 antibody as monotherapy or in combination therapy, as well as a series of treatments administered in a planned manner. In one embodiment, the prior treatment was CD38 antibody monotherapy. In one embodiment, the prior treatment was a combination therapy comprising a CD38 antibody. For example, the prior treatment may have been a CD38 antibody in combination with a proteasome inhibitor (PI) and an immunomodulator. In some embodiments, the CD38 antibody is daratumumab or isatuximab.

[0205] In some cases, the target population may also be patients for whom daratumumab and / or isatuximab as monotherapy have limited efficacy.

[0206] In some aspects, hematological malignancies can be characterized as cancers that are “refractory” or “recurrent” to prior treatment. In further aspects, prior treatment may include one or more of PIs, IMiDs, and CD38 antibodies, for example, the CD38 antibody being daratumumab or isatuximab. Typically, this indicates that prior treatment did not achieve a complete response (CR), e.g., the cancer was unresponsive to CD38 antibody monotherapy or combination therapy, or the cancer progressed within a predetermined period after the completion of CD38 antibody therapy. Examples of such combination therapies include, but are not limited to, combinations of CD38 antibodies with PIs or IMiDs, or combinations of PIs and IMiDs. Similarly, it may indicate that prior treatment did not achieve a complete response (CR), e.g., the cancer was unresponsive to a PI, an IMiD, or a combination of them, or the cancer progressed within a predetermined period after the completion of such treatment. A person skilled in the art can determine whether a cancer is refractory to previous treatments based on what is known in the art, including the available guidelines for each type of cancer.

[0207] For example, in multiple myeloma, refractory and relapsing disease can be identified according to the guidelines published by Rajkumar, Harousseau, et al. on behalf of the International Myeloma Workshop Consensus Panel, Consensus recommendations for the uniform reporting of clinical trials: report of the International Myeloma Workshop Consensus Panel, Blood 2011;117:4691-4695.

[0208] Refractory myeloma can be defined as a disease that is unresponsive during first-line therapy or salvage therapy, or a disease that progresses within 60 days of the last treatment. Unresponsiveness is defined as failure to achieve minimal response during treatment or the occurrence of disease progression (PD). There may be two categories of refractory myeloma: relapsed / refractory myeloma and primary refractory myeloma.

[0209] Relapsed or refractory myeloma can be defined as a disease that is unresponsive to salvage therapy, or a disease that progresses within 60 days of the last treatment in patients who previously achieved minimum response (MR) or better at some point in time but have since progressed during the course of the disease.

[0210] Primary refractory myeloma can be defined as a disease that is unresponsive in patients who have never achieved a minimum response or better with any treatment. This includes patients who have never achieved a minimum response or better, who have no significant changes in M ​​protein and no evidence of clinical progression, and primary refractory PD in patients who meet the criteria for true PD. When reporting the treatment efficacy in patients with primary refractory myeloma, the efficacy in these two subgroups ("unresponsive / non-progressive" and "progressive") should be specified separately.

[0211] Relapsed myeloma can be defined as a myeloma that has been treated previously, has progressed, and requires the initiation of salvage therapy, but does not meet the criteria for either "primary refractory myeloma" or "relapsed / refractory myeloma."

[0212] For details on specific responses (CR, PR, etc.) in multiple myeloma and the methods used to evaluate them, please refer to Rajkumar, Harousseau et al., 2011 (cited above).

[0213] Thus, in some embodiments, an antibody or a pharmaceutical composition comprising the antibody according to any aspect or embodiment herein is for use in treating cancer refractory to a previous treatment comprising one or more of a PI, an IMiD, and a CD38 antibody. In one embodiment, the previous treatment comprises a CD38 antibody. In certain embodiments, the cancer is identified as a refractory cancer prior to use.

[0214] In another embodiment, provided herein is a method for treating cancer in a subject, comprising the following steps: 1. identifying the subject as refractory to a previous treatment comprising one or more of a PI, an IMiD, and a CD38 antibody, and 2. administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising an antibody or an antibody variant according to any aspect or embodiment herein.

[0215] In one embodiment, the previous treatment comprises a CD38 antibody.

[0216] In another embodiment, provided is a method for treating cancer refractory to a previous treatment comprising one or more of a PI, an IMiD, and a CD38 antibody in a subject, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising an antibody or the antibody according to any aspect or embodiment herein. In one embodiment, the previous treatment comprises a CD38 antibody.

[0217] In some embodiments, the PI is selected from the group consisting of bortezomib, carfilzomib, and ixazomib.

[0218] In some embodiments, the IMiD is selected from the group consisting of thalidomide, lenalidomide, and pomalidomide.

[0219] In some embodiments, the CD38 antibody is daratumumab. In some embodiments, the CD38 antibody is isatuximab. In some embodiments, the CD38 antibody may be felzartamab or mezartamab.

[0220] In some embodiments, an antibody or a pharmaceutical composition comprising the antibody according to any aspect or embodiment herein is for use in treating cancer that has recurred after a previous treatment comprising one or more of a PI, an IMiD, and a CD38 antibody. In one embodiment, the previous treatment comprises a CD38 antibody. In certain embodiments, the cancer is identified as recurrent prior to use.

[0221] In another embodiment, provided herein is a method for treating cancer in a subject, comprising the following steps: 1. Identifying a subject as having recurred after a previous treatment comprising one or more of a PI, an IMiD, and a CD38 antibody, and 2. Administering to the subject a therapeutically effective amount of an antibody or a pharmaceutical composition comprising the antibody according to any aspect or embodiment herein.

[0222] In one embodiment, the previous treatment comprises a CD38 antibody.

[0223] In another embodiment, provided herein is a method for treating cancer that has recurred after a previous treatment comprising one or more of a PI, an IMiD, and a CD38 antibody in a subject, comprising the step of administering to the subject a therapeutically effective amount of an antibody or a pharmaceutical composition comprising the antibody according to any aspect or embodiment herein. In one embodiment, the previous treatment comprises a CD38 antibody.

[0224] In some embodiments, the PI is selected from the group consisting of bortezomib, carfilzomib, and ixazomib.

[0225] In some embodiments, the IMiD is selected from the group consisting of thalidomide, lenalidomide, and pomalidomide.

[0226] In some embodiments, the CD38 antibody is daratumumab. In some embodiments, the CD38 antibody is isatuximab. In some embodiments, the CD38 antibody may be felzartamab or mezartamab.

[0227] In certain embodiments, the antibody according to the present invention is administered in a therapeutically effective dose and / or for a sufficient duration to treat refractory or recurrent cancer.

[0228] In some aspects of the methods and uses described herein, the refractory or recurrent cancer is a hematological cancer.

[0229] In some embodiments, refractory or recurrent cancers are selected from the group consisting of multiple myeloma (MM), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), mantle cell lymphoma (MCL), follicular lymphoma (FL), and diffuse large B-cell lymphoma (DLBCL).

[0230] In some embodiments, refractory or recurrent cancers are selected from the group consisting of multiple myeloma (MM), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), and follicular lymphoma (FL).

[0231] In some cases, refractory or recurrent cancer is chronic lymphocytic leukemia (CLL).

[0232] In some cases, refractory or recurrent cancer is mantle cell lymphoma (MCL).

[0233] In some cases, refractory or recurrent cancer is diffuse large B-cell lymphoma (DLBCL).

[0234] In some cases, refractory or recurrent cancer is follicular lymphoma (FL).

[0235] In some particularly preferred embodiments, the refractory or recurrent cancer is multiple myeloma (MM).

[0236] In some embodiments, relapsed or refractory multiple myeloma is characterized by evidence of disease progression in the subject to the most recent prior treatment regimen, based on the IMWG 2016 criteria with measurable lesions. The criteria may be selected from the following group: a. A previous record of monoclonal plasma cells in ≥10% of the bone marrow, or the presence of a plasmacytoma confirmed by biopsy; b. IgG, IgA, IgD, or IgM myeloma: Serum M protein level ≥ 0.5 g / dL (≥ 5 g / L) or urinary M protein level ≥ 200 mg / 24 hours; c. Light chain myeloma: Serum Ig free light chain (FLC) ≥ 10 mg / dL and abnormal serum IgκλFLC ratio.

[0237] In one embodiment, the criteria for judgment are a combination of a. and b. or c. as described above.

[0238] In some preferred embodiments, by any of the methods and uses herein, the hematological malignancy is diffuse large B-cell lymphoma (DLBLC), such as relapsed or refractory DLBCL. Thus, in another aspect, the method according to the invention provided in any of the embodiments described herein treats DLBCL, such as refractory or relapsed DLBLC, such as DLBCL that is refractory or relapsed to previous treatment (e.g., daratumumab or isatuximab) against CD38, in a (human) subject that needs it. The dosage and treatment cycles can be as described elsewhere herein. Thus, in any of these embodiments, the amount of antibody administered (per dose and / or per treatment cycle) is at least (about) 4 mg / kg body weight, such as 4 mg / kg to 24 mg / kg body weight. Preferably, the antibody is administered at a dose of (about) 8 mg / kg body weight to (about) 24 mg / kg, or at a dose of (about) 8 mg / kg body weight to (about) 16 mg / kg body weight. In one embodiment of the treatment of DLBCL, the antibody is administered at a dose of (about) 4 mg / kg body weight. In another embodiment, the antibody is administered at a dose of (about) 8 mg / kg body weight. In yet another embodiment, the antibody is administered at a dose of (about) 16 mg / kg body weight.

[0239] In one embodiment, relapsed or refractory DLBCL includes both de novo DLBCL or histologically transformed DLBCL. A relapsed disease can be defined as the reappearance or growth of lymphoma after at least a 6-month duration of response (DOR). A refractory disease can be defined as the failure to achieve a response after at least 2 cycles of therapy, or as a reappearance after <6 months of DOR.

[0240] The evaluation of the effectiveness of the treatment against DLBCL can be carried out according to the Lugano response criteria for lymphoma (Cheson et al., 2014), as shown in Tables 2 and 3 below.

[0241] (Table 2) Lugano response criteria for DLBCL - CT / MRI scan TIFF2026513656000004.tif112161

[0242] (Table 3) Lugano efficacy evaluation criteria DLBCL-PET / CT scan TIFF2026513656000005.tif85167a. PET 5PS: 1. No uptake exceeding background; 2. Uptake ≤ mediastinum; 3. Uptake > mediastinum, but ≤ liver; 4. Uptake slightly higher than in the liver; 5. Uptake significantly higher than in the liver and / or new lesions; X. New uptake sites unlikely to be associated with lymphoma.

[0243] Therapeutic effect effect In any of the methods and uses described herein, administration of the antibody according to the present invention may induce and / or improve one or more therapeutic effects in a treated subject compared to baseline.

[0244] Therefore, in some embodiments, the one or more therapeutic effects induced and / or improved are selected from the group consisting of overall response rate, duration of response, and time to response.

[0245] In some embodiments, the therapeutic effect is defined as a strict complete response, complete response, best partial response, partial response, minimal response, or stable state. Optionally, treatment may be continued until the disease progresses or the patient no longer receives any benefit.

[0246] In one embodiment, the therapeutic effect is a strict complete response.

[0247] In one embodiment, the therapeutic effect is a complete response.

[0248] In one embodiment, the therapeutic effect is the best partial response.

[0249] In one embodiment, the therapeutic effect is a partial response.

[0250] In one embodiment, the therapeutic effect is minimal response.

[0251] In one embodiment, the therapeutic effect remains stable.

[0252] In one embodiment, the hematological malignancy is preferably (relapsed or refractory) multiple myeloma, the therapeutic effect is an overall response rate of at least (about) 14% of the treated subjects, for example, at least (about) 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 35%, 40%, or higher, and optionally, the antibody is administered in a dose of at least (about) 4 mg / kg, for example, at about 4-24 mg / kg.

[0253] In some embodiments, the hematological malignancy is a cancer that has not been previously treated with a prior treatment comprising a CD38 antibody, preferably daratumumab or isatuximab, preferably (relapsed or refractory) multiple myeloma, the therapeutic effect is an overall response rate of at least (about) 20% of the treated subjects, e.g., at least (about) 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or higher, and optionally, the antibody is administered in a dose of at least (about) 4 mg / kg, e.g., at least 4 mg / kg, e.g., 4 to 24 mg / kg, e.g., e.g., 16 mg / kg.

[0254] In some embodiments, the hematological malignancies are cancers that are relapsed or refractory to prior anticancer therapies, e.g., cancers that are relapsed or refractory to prior therapies, e.g., cancers that are (relapsed or refractory) multiple myeloma, e.g., (relapsed or refractory) multiple myeloma, and the therapeutic effect is an overall response rate of at least (about) 6% of the treated subjects, e.g., at least (about) 8%, 10%, 12%, 14%, 16%, 18%, 20%, 25%, 30%, 35%, 40%, or higher, and optionally, the antibody is administered in a dose of at least (about) 4 mg / kg, e.g., at least (about) 4 mg / kg, e.g., at least (about) 4 to 24 mg / kg, e.g., at least (about) 16 mg / kg.

[0255] In a particular preferred embodiment, the hematological malignancy is a cancer that has not been previously treated with a prior treatment comprising a CD38 antibody, preferably daratumumab or isatuximab, preferably (relapsed or refractory) multiple myeloma, and the therapeutic effect is at least (about) 25 best partial responses (VGPRs) in the treated subjects, e.g., at least (about) 30%, 35%, 40%, 45%, 50%, 60%, 70% or higher VGPRs in the treated subjects, and optionally, the dose is at least (about) 4 mg / kg body weight, or alternatively at least (about) 8 mg / kg body weight, or at least (about) 16 mg / kg body weight, or at least (about) 24 mg / kg body weight. In certain embodiments, the therapeutic effect is a complete response (CR) of at least 9% in the treated subjects, e.g., at least 10%, 15%, 20%, 24%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, or higher, and optionally, the dose is at least 4 mg / kg body weight, or alternatively at least 8 mg / kg body weight, or at least 16 mg / kg body weight, or at least 24 mg / kg body weight.

[0256] In some embodiments, the hematological malignancy is a cancer that is recurrent or refractory to prior anticancer therapies, e.g., prior therapies comprising a CD38 antibody, preferably daratumumab or isatuximab, and the therapeutic effect is a partial response (PR) of at least 6% of the treated subjects, e.g., at least 8%, 10%, 12%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25% or higher PR in the treated subjects, and optionally, the dose is at least about 4 mg / kg body weight, or alternatively at least about 8 mg / kg body weight, or at least about 16 mg / kg body weight, or at least about 24 mg / kg body weight, preferably at least 16 mg / kg body weight.

[0257] In one embodiment, the antibody, when used in any aspect or embodiment disclosed herein, improves the one or more therapeutic effects in the subject compared to treatment with a control antibody (in a similar, equivalent, or equivalent dose). The control may be a reference antibody having an amino acid sequence (typically heavy and light chain amino acid sequences) identical to that of the antibody of the present invention, except for one or more mutations at E430, E345, and / or S440. In a preferred embodiment, the antibody of the present invention contains a mutation at position E430, preferably E340G (in the human IgG1 heavy chain), and the reference antibody does not contain the mutation at position E430 (i.e., is wild-type at that position), and preferably contains a wild-type CH3 / Fc region (in the human IgG1 heavy chain).

[0258] In another embodiment, the control is a reference antibody having the same amino acid sequence (typically heavy and light chain amino acid sequences) as the antibody of the present invention, except for different VH and VL sequences.

[0259] In one embodiment, the antibody, when used in any aspect or embodiment disclosed herein, improves the one or more therapeutic effects in the subject compared to a reference antibody, wherein the reference antibody comprises the VH and VL region sequences of antibody C, i.e., SEQ ID NO:1 and SEQ ID NO:5, respectively, and the CH and CL region sequences, which are identical to the antibody of the present invention except for one or more mutations at E430, E345, and / or S440. In a preferred embodiment, the antibody of the present invention comprises a mutation at position E430 (in the human IgG1 heavy chain), preferably E340G, and the reference antibody does not comprise the mutation at position E430 (i.e., is wild-type at that position), preferably comprising a wild-type CH3 / Fc region (in the human IgG1 heavy chain).

[0260] In some embodiments, the antibody, when used in accordance with any aspect or embodiment disclosed herein, improves one or more therapeutic effects in the subject compared to a reference antibody, wherein the reference antibody comprises the VH and VL region sequences of antibody B, i.e., SEQ ID NO:8 and SEQ ID NO:9, respectively, and the CH and CL region sequences which are identical to those of the antibody of the present invention.

[0261] In one embodiment, the antibody, when used in any aspect or embodiment disclosed herein, improves the one or more therapeutic effects in the subject compared to a reference antibody, wherein the reference antibody comprises the VH and VL region sequences of antibody B, i.e., SEQ ID NO:8 and SEQ ID NO:9, respectively, and the CH and CL region sequences of SEQ ID NO:20(IgGm(f)) and SEQ ID NO:37(κ), respectively, or the reference antibody is antibody B, daratumumab, or isatuximab. In a preferred embodiment, the antibody of the present invention comprises a mutation at position E430 (in the human IgG monohelic acid), preferably E340G.

[0262] The reference antibody may be ferzaltamab or mezaditamab.

[0263] In one embodiment, the one or more therapeutic effects can be achieved when the antibody is administered in a dose of at least (about) 4 mg / kg body weight, for example, 4 mg to (about) 24 mg / kg body weight, or at any intermediate dose level or range described herein.

[0264] In one embodiment, the one or more therapeutic effects can be achieved with a dose of at least (about) 8 mg / kg body weight, for example, (about) 8 to (about) 24 mg / kg body weight or (about) 8 to (about) 16 mg / kg body weight.

[0265] In one embodiment, the one or more therapeutic effects can be achieved with a dose of approximately 4 mg / kg body weight.

[0266] In one embodiment, the one or more therapeutic effects can be achieved with a dose of approximately 8 mg / kg body weight.

[0267] In one embodiment, the one or more therapeutic effects can be achieved with a dose of approximately 16 mg / kg body weight.

[0268] In one embodiment, the one or more therapeutic effects can be achieved with a dose of approximately 24 mg / kg body weight.

[0269] In one embodiment, the one or more therapeutic effects may be achieved when the antibody is administered to a subject who has not previously been treated with an anti-CD38 antibody, such as daratumumab, isatuximab, ferzaltamab, and mezaditamab (an anti-CD38 mAb naive subject).

[0270] In one embodiment, the one or more therapeutic effects may be achieved when the antibody is administered to a subject who has previously been treated with an anti-CD38 antibody, for example, a subject with hematological malignancies whose cancer has relapsed or become refractory after prior treatment including an anti-CD38 antibody such as daratumumab, isatuximab, ferzaltamab, and mezaditamab (a subject with prior anti-CD38 treatment).

[0271] Management of side effects In some embodiments of any of the methods and uses of the present invention, the subject may be treated for the management of infusion reactions (IRRs). For this purpose, pre-infusion drug therapy (e.g., corticosteroids, antipyretics, antihistamines, leukotriene receptor antagonists) and / or post-infusion drug therapy (e.g., corticosteroids) may be administered to reduce the risk of IRRs, particularly grade 2 or higher IRRs, or for the treatment of IRRs. The pre-infusion drug therapy may be administered about 1 to 3 hours before the administration of the antibody, and / or the post-infusion drug therapy may be administered 2 days after the administration of the antibody.

[0272] Pre-infusion drug therapy may include corticosteroids (e.g., methylprednisolone, betamethasone, dexamethasone, triamcinolone, prednisone and / or prednisolone), antihistamines (e.g., diphenhydramine), antipyretics (e.g., paracetamol), and / or leukotriene receptor antagonists (e.g., montelukast), optionally, a. The corticosteroid is administered in an amount of approximately 60-100 mg of methylprednisolone or an equivalent dose; b. The diphenhydramine is administered in doses of approximately 25-50 mg; c. The paracetamol is administered in doses of approximately 650–1000 mg; and / or d. The montelukast is administered in a dose of approximately 10 mg.

[0273] Post-infusion drug therapy may include corticosteroids, such as methylprednisolone, betamethasone, dexamethasone, triamcinolone, prednisone, and / or prednisolone, and optionally, the corticosteroid is administered in a dose of 20 mg methylprednisolone or an equivalent dose.

[0274] In some embodiments of any of the methods and uses of the present invention, the subject may be treated for the management of cytopenia, e.g., neutropenia or thrombocytopenia, particularly grade 3 or 4 neutropenia or thrombocytopenia. Such treatment may be by transfusion of granulocyte colony-stimulating factor (G-CSF) and / or other blood growth factors (e.g., erythropoietin) and / or blood products (e.g., transfusion of red blood cells and platelets), preferably by G-CSF. In one embodiment, G-CSF is administered when the antibody is administered at a dose level of 4 mg / kg body weight or higher, for example, when grade 3 or 4 neutropenia is observed.

[0275] Pharmacokinetics In one embodiment, the antibody exhibits higher clearance than the reference antibody when used in accordance with the methods and uses described herein.

[0276] In one embodiment, the reference antibody is a normal IgG antibody, for example, an IgG1 antibody that does not contain mutations at one or more amino acid residues selected from the group corresponding to E430, E345, and S440 in the human IgG1 heavy chain. In a preferred embodiment, the antibody of the present invention contains a mutation at position E430, preferably E340G, and the reference antibody does not contain the mutation at position E430 (i.e., is wild-type at that position).

[0277] Accordingly, in one embodiment, the antibody exhibits higher clearance than the reference antibody when used in accordance with the methods and uses described herein, wherein the reference antibody comprises the VH and VL region sequences of antibody C, i.e., SEQ ID NO:1 and SEQ ID NO:5, respectively, and the CH and CL regions which are identical except for one or more mutations at E430, E345 and / or S440. In a preferred embodiment, the antibody of the present invention comprises a mutation at position E430 (in the human IgG1 heavy chain), preferably E340G, and the reference antibody does not contain the mutation at position E430 (i.e., is wild-type at the position), preferably comprising a wild-type CH3 / Fc region (in the human IgG1 heavy chain).

[0278] In one embodiment, the antibody exhibits higher clearance than the reference antibody when used in accordance with the methods and uses described herein, wherein the reference antibody is another anti-CD38 antibody that does not contain mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 in the human IgG1 heavy chain. In a preferred embodiment, the antibody of the present invention contains a mutation at position E430 (in the human IgG1 heavy chain), preferably E340G, and the reference antibody does not contain the mutation at position E430 (i.e., is wild-type at the position), preferably containing a wild-type CH3 / Fc region (in the human IgG1 heavy chain).

[0279] Therefore, in one embodiment, the antibody exhibits higher clearance than the reference antibody when used according to the methods and uses described herein, wherein the reference antibody contains the VH and VL region sequences of antibody B, i.e., SEQ ID NO:8 and SEQ ID NO:9, respectively, and the CH and CL region sequences of SEQ ID NO:20(IgGm(f)) and SEQ ID NO:37(κ), respectively, or the reference antibody is antibody B, or the reference antibody is daratumumab or isatuximab. In a preferred embodiment, the antibody of the present invention contains a mutation at position E430 (in the human IgG monohelic acid), preferably E340G.

[0280] The reference antibody may be ferzaltamab or mezaditamab.

[0281] In particular, higher clearance than that of the reference antibody may occur when the antibody is administered at doses of at least (approximately) 4 mg / kg body weight, e.g., (approximately) 6 mg / kg body weight, e.g., (approximately) 8 mg / kg body weight, e.g., (approximately) 10 mg / kg body weight, e.g., (approximately) 12 mg / kg body weight, e.g., (approximately) 14 mg / kg body weight, e.g., (approximately) 16 mg / kg body weight, e.g., (approximately) 20 mg / kg body weight, e.g., (approximately) 22 mg / kg body weight, e.g., (approximately) 24 mg / kg body weight. At such high doses, target-mediated drug elimination appeared to be saturated.

[0282] In one embodiment, the clearance is a non-target-mediated clearance.

[0283] In one embodiment, the higher clearance is FcRn-dependent.

[0284] As used herein, clearance is defined as the dose divided by the estimated area under the serum concentration-time curve from the start of administration to indefinite time.

[0285] Pharmacodynamics When used in accordance with the methods and uses described herein, antibodies may typically have one or more of the following effects on a subject: activation of the complement system, depletion of peripheral blood NK cells, expansion and proliferation of peripheral blood NK cells, or any combination thereof.

[0286] When used in accordance with the methods and uses described herein, antibodies typically do not have, or substantially do not have, one or more of the following effects in a subject: a dose-dependent increase in plasma levels of pro-inflammatory cytokines, or a dose-dependent reduction of certain non-tumor cells other than NK cells (B cells, T cells, monocytes and / or NKT-like cells), or any combination thereof.

[0287] In one embodiment, the antibody, when used in accordance with the methods and uses described herein, induces complement system activation in a subject. This complement system activation may be, for example, a reduction in complement component C2, as measured in peripheral blood. The reduction in C2 indicates the CDC activity of the antibody of the present invention in vivo. The reduction in C2 may be transient after each dose, typically returning to baseline within 8 days post-dose, for example, before the next dose. In one embodiment, the C2 level may decrease by at least 30% from baseline (mean peak or median peak), for example, at least (about) 35%, 40%, 45%, 50%, 55%, 58%, 60%, 64%, 70%, 75%, 80%, or more.

[0288] Peripheral blood C2 levels can be measured, for example, by any suitable method known in the art, essentially by the method described in Example 6.

[0289] The activation of the complement system may also be a reduction in the total complement lysis activity (CH50), as measured, for example, in peripheral blood. A reduction in CH50 indicates the CDC activity of the antibody of the present invention in vivo. The reduction in CH50 may be transient after each dose and return to baseline, for example, before the next dose, typically within 8 days post-dose. In one embodiment, the CH50 level may decrease by more than 20% from baseline (mean peak or median peak), for example, by (about) 25%, 30%, 32%, 35%, 40%, 45%, 48%, 50%, 53%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or even more.

[0290] The CH50 level in peripheral blood can be measured, for example, by any suitable method known in the art, essentially by the method described in Example 7.

[0291] In some embodiments, the complement system activation of the antibody of the present invention, e.g., reduction of C2 and / or CH50, is greater than that of the control antibody. The control may be a reference antibody having the same amino acid sequence (typically heavy and light chain amino acid sequences) as the antibody, except for one or more mutations at, for example, E430, E345, and / or S440. In a preferred embodiment, the antibody of the present invention contains a mutation at position E430 (in the human IgG1 heavy chain), preferably E340G, and the reference antibody does not contain the mutation at position E430 (i.e., is wild-type at that position), preferably containing a wild-type CH3 / Fc region (in the human IgG1 heavy chain). Alternatively, the control may be a reference antibody having the same amino acid sequence (typically heavy and light chain amino acid sequences) as the antibody of the present invention, except for different VH and VL sequences. Such a reference antibody may instead have the VH and VL sequences of antibody B or A shown in Table 4, for example. Preferably, the VH and VL sequences of the reference antibody are those of antibody B. Alternatively, the reference antibody may be an antibody that binds to the same target but has a different sequence.

[0292] Accordingly, in one embodiment, the antibody, when used in any aspect or embodiment disclosed herein, induces a higher complement system activation than the reference antibody, for example, a significant reduction in C2 and / or CH50, wherein the reference antibody comprises the same CH and CL region sequences as the antibody of the present invention, except for one or more mutations at E430, E345 and / or S440, i.e., SEQ ID NO:1 and SEQ ID NO:5, respectively. In a preferred embodiment, the antibody of the present invention comprises a mutation at position E430 (in the human IgG1 heavy chain), preferably E340G, wherein the reference antibody does not contain the mutation at position E430 (i.e., is wild-type at that position), and preferably comprises a wild-type CH3 / Fc region (in the human IgG1 heavy chain).

[0293] In another embodiment, the antibody, when used in any aspect or embodiment disclosed herein, induces a higher complement system activation than the reference antibody, e.g., a greater reduction of C2 and / or CH50, wherein the reference antibody comprises the VH and VL region sequences of antibody C, i.e., SEQ ID NO:1 and SEQ ID NO:5, respectively, and the CH and CL region sequences of SEQ ID NO:20(IgGm(f)) and SEQ ID NO:37(κ), respectively.

[0294] In another embodiment, when the antibody is used according to any aspect or embodiment disclosed herein, it induces a higher complement system activation than the reference antibody, e.g., a greater reduction of C2 and / or CH50, wherein the reference antibody includes the VH and VL region sequences of antibody B, i.e., SEQ ID NO:8 and SEQ ID NO:9, respectively, as well as the same CH and CL region sequences as the antibody of the present invention.

[0295] In one embodiment, the antibody, when used according to any aspect or embodiment disclosed herein, induces a higher complement system activation than the reference antibody, e.g., a greater reduction of C2 and / or CH50, wherein the reference antibody includes the VH and VL region sequences of antibody B, i.e., SEQ ID NO:8 and SEQ ID NO:9, respectively, and the CH and CL region sequences of SEQ ID NO:20(IgGm(f)) and SEQ ID NO:37(κ), respectively, or the reference antibody is antibody B. In a preferred embodiment, the antibody of the present invention includes a mutation at position E430 (in the human IgG1 heavy chain), preferably E340G.

[0296] In one embodiment, the antibody does not deplete the complement system when used in accordance with the methods and uses described herein. Therefore, the reduction in C2 and / or CH50 is preferably transient and returns to baseline after a certain period of time, preferably before the next dose is administered, for example, within (about) 8 days after administration. Preferably, the complement system is not depleted at any dose level between 4 and 24 mg / kg body weight, for example, when the antibody is administered at doses of (about) 4 mg / kg body weight, (about) 6 mg / kg body weight, (about) 8 mg / kg body weight, (about) 10 mg / kg body weight, (about) 12 mg / kg body weight, (about) 14 mg / kg body weight, (about) 16 mg / kg body weight, (about) 20 mg / kg body weight, (about) 22 mg / kg body weight, and (about) 24 mg / kg body weight. Therefore, at any of these dose levels, the CH2 and / or CH50 levels preferably return to baseline after a few days, for example, within (about) 8 days.

[0297] In another embodiment, the antibody induces depletion of peripheral blood NK cells in a subject when used in accordance with the methods and uses described herein. NK cell depletion indicates the ADCC activity of the antibody of the present invention in vivo. In one embodiment, NK cell deficiency is induced when the antibody is administered at dose levels of at least 0.2 mg / kg body weight, e.g., 0.2 to 24 mg / kg, e.g., approximately 4 mg / kg body weight, approximately 6 mg / kg body weight, approximately 8 mg / kg body weight, approximately 10 mg / kg body weight, approximately 12 mg / kg body weight, approximately 14 mg / kg body weight, approximately 16 mg / kg body weight, approximately 20 mg / kg body weight, approximately 22 mg / kg body weight, and approximately 24 mg / kg body weight. In one embodiment, NK cells remained reduced or substantially reduced throughout the entire treatment, for example, throughout Cycle 1, or additionally throughout Cycle 2, or additionally throughout Cycle 3, or additionally throughout Cycle 4, or additionally throughout Cycle 5, and optionally thereafter.

[0298] The number of peripheral blood NK cells can be determined, for example, by any suitable method known in the art, for example, essentially by the method described in Example 5.

[0299] In another embodiment, the antibody, when used in accordance with the methods and uses described herein, induces the expansion and proliferation of peripheral blood T cells in a subject. This T cell expansion and proliferation demonstrates the immunomodulatory activity of the antibody of the present invention in vivo (e.g., inhibition of CD38 cyclase activity or depletion of regulatory cells such as Tregs). In one embodiment, the subject has not received prior treatment including a CD38 antibody (e.g., daratumumab and / or isatuximab).

[0300] The number of peripheral blood T cells can be determined, for example, by any suitable method known in the art, for example, essentially by the method described in Example 5.

[0301] In another embodiment, when the antibody is used in accordance with the methods and uses described herein, it does not induce or substantially induces a dose-dependent increase in plasma levels of pro-inflammatory cytokines, such as IL-2, IL-6, IL-8, IL-10, IFNγ, and / or TNFα, in the subject. In another embodiment, when the antibody is used in accordance with the methods and uses described herein, it does not induce or substantially induces a dose-dependent increase in plasma levels of pro-inflammatory cytokines, such as IL-2, IL-6, IL-8, IL-10, and / or IFNγ, in the subject.

[0302] In another embodiment, when the antibody is used in accordance with the methods and uses described herein, it does not induce or substantially induces a dose-dependent reduction of certain CD38-expressing non-tumor cells (other than NK cells), the CD38-expressing non-tumor cells being selected from the group consisting of B cells, T cells, monocytes and / or NKT-like cells in the subject.

[0303] The number of B cells, T cells, monocytes, and NKT-like cells can be determined, for example, by any suitable method known in the art, for example, essentially by the method described in Example 5.

[0304] In one embodiment, when the antibody is used in accordance with the methods and uses described herein, a. The subject has an inhibitory effect on CD38 cyclase activity; b. In the subjects described above, induce complement-dependent cell-mediated cytotoxicity (CDC) in cells expressing human CD38; c. In the subjects described above, antibody-dependent cell-mediated injury (ADCC) is induced in cells expressing human CD38; d. In the subjects described above, induce antibody-dependent cell phagocytosis (ADCP) in cells expressing human CD38; e. In the subject, induce apoptosis in the presence of FcgR-containing cells; f. Induce trogocytosis in cells expressing human CD38; or Any combination of ga~f.

[0305] In some embodiments, one or all of a., b., and f. are higher (when administered at similar, equivalent, or comparable doses) compared to a reference antibody that does not contain mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 (amino acid residues are numbered according to the EU index) in the human IgG monohelic acid single heavy chain.

[0306] Appropriate assays for evaluating CD38 cyclase activity, CDC, ADCC, ADCP, trogocytosis, and apoptosis are known in the art and are described, for example, in WO 2020 / 012036 A1 and WO 2020 / 012038 A1.

[0307] In one embodiment, the antibody, when used in accordance with the methods and uses described herein, induces trogocytosis in the subject, for example, trogocytosis of CD38 from donor CD38-expressing cells to acceptor cells. Typical acceptor cells include T cells and B cells, monocytes / macrophages, dendritic cells, neutrophils, and NK cells. Preferably, the acceptor cells are lymphocytes expressing the Fc gamma (Fcγ) receptor, such as macrophages or PBMCs. In particular, the antibody of the present invention may mediate an increase in trogocytosis compared to a control. The control may be a reference antibody having the same amino acid sequence (typically heavy and light chain amino acid sequences) as the antibody of the present invention, except for one or more mutations at E430, E345, and / or S440. In a preferred embodiment, the antibody of the present invention comprises a mutation at position E430, preferably E340G (in the human IgG1 heavy chain), and the reference antibody does not comprise the mutation at position E430 (i.e., is wild-type at that position), and preferably comprises a wild-type CH3 / Fc region (in the human IgG1 heavy chain).

[0308] In another embodiment, the control is a reference antibody having an amino acid sequence (typically heavy and light chain amino acid sequences) identical to that of the antibody of the present invention, except for different VH and VL sequences.

[0309] Appropriate assays for evaluating trogocytosis are known in the art and are described, for example, in WO 2020 / 012036 A1 (Genmab A / S) and WO 2020 / 012038 A1 (Genmab A / S).

[0310] In one embodiment, the antibody, when used according to any aspect or embodiment disclosed herein, induces trogocytosis of target cells expressing a higher level of CD38 than the reference antibody, wherein the reference antibody comprises the VH and VL region sequences of antibody C, i.e., SEQ ID NO:1 and SEQ ID NO:5, respectively, and the same CH and CL region sequences as the antibody of the present invention, except for one or more mutations at E430, E345, and / or S440. In a preferred embodiment, the antibody of the present invention comprises a mutation at position E430 (in the human IgG1 heavy chain), preferably E340G, and the reference antibody does not comprise the mutation at position E430 (i.e., is wild-type at that position), preferably comprising a wild-type CH3 / Fc region (in the human IgG1 heavy chain).

[0311] In some embodiments, the antibody, when used according to any aspect or embodiment disclosed herein, induces trogocytosis of target cells expressing a higher level of CD38 than the reference antibody, wherein the reference antibody comprises the VH and VL region sequences of antibody B, i.e., SEQ ID NO:8 and SEQ ID NO:9, respectively, as well as the CH and CL region sequences identical to those of the antibody of the present invention.

[0312] In one embodiment, the antibody, when used according to any aspect or embodiment disclosed herein, induces trogocytosis of target cells expressing a higher level of CD38 than the reference antibody, wherein the reference antibody comprises the VH and VL region sequences of antibody B, i.e., SEQ ID NO:8 and SEQ ID NO:9, respectively, and the CH and CL region sequences of SEQ ID NO:20(IgGm(f)) and SEQ ID NO:37(κ), respectively, or the reference antibody is antibody B. In a preferred embodiment, the antibody of the present invention comprises a mutation at position E430 (in the human IgG1 heavy chain), preferably E340G.

[0313] In one embodiment, the antibody, when used in any aspect or embodiment disclosed herein, induces a reduction in the trogocytosis-mediated activity of CD38 on CD38-expressing tumor cells in the subject. In one embodiment, the antibody, when used in any aspect or embodiment disclosed herein, induces a reduction in the trogocytosis-mediated activity of CD38 on CD38-expressing immune cells in the subject.

[0314] In one embodiment, the CD38-expressing immune cells are cells, and are CD38-expressing immunosuppressive cells.

[0315] In one embodiment, reducing the trogocytosis-mediated activity of CD38 on CD38-expressing immunosuppressive cells reduces their immunosuppressive activity.

[0316] In one embodiment, the CD38-expressing immunosuppressive cells include regulatory T cells (Treg), regulatory B cells (Breg), myeloid-derived suppressor cells (MDSC), immunosuppressive NK cells, immunosuppressive NKT cells, immunosuppressive antigen-expressing cells (APC), immunosuppressive macrophages, or any combination of two or more thereof.

[0317] In one embodiment, the immune cells are Tregs, and therefore the antibody induces a reduction in the trogocytosis-mediated activity of CD38 on CD38-expressing Tregs.

[0318] antigen-binding region and variable region In the methods and uses of the present invention, the antigen-binding region of the antibody includes one or more antibody-variable domains, e.g., a VH region and a VL region, that enable specific binding to CD38. Similarly, the heavy chain and light chain each include a VH region and a VL region. Hereinafter, references to sequences within the antigen-binding region may also apply to sequences of the heavy chain and / or light chain of the antibody according to the present invention. Advantageously, the CDR, VH region and / or VL region are similar to or identical to those of antibody C described in Table 4.

[0319] In one preferred embodiment, the antigen-binding region, and / or the heavy and / or light chain, comprises the CDR of antibody C as described in SEQ ID NO:2(VH-3003-C_CDR1), SEQ ID NO:3(VH-3003-C_CDR2), SEQ ID NO:4(VH-3003-C_CDR3), SEQ ID NO:6(VL-3003-C_CDR1), AAS(VL-3003-C_CDR2), and SEQ ID NO:7(VL-3003-C_CDR3). In another preferred embodiment, the VH and VL sequences are those of antibody C, i.e., the VH region comprises the sequence of SEQ ID NO:1 (VH-3003-C) and the VL region comprises the sequence of SEQ ID NO:5 (VL-3003-C).

[0320] However, it is well known in the art that mutations can be made in the VH and VL regions of an antibody, for example, to increase the antibody's affinity for its target antigen, to decrease its potential immunogenicity, and / or to increase the yield of the antibody expressed by host cells. Accordingly, in some embodiments, antibodies are also intended to include variants of the CDR, VH, and / or VL sequences of antibody C, in particular functional variants of the VL and / or VH regions of antibody C. The functional variant may differ from the parent VH and / or VL sequence by, for example, one or more amino acids in one or more CDRs, but the antigen-binding region still retains at least a significant percentage (at least about 50%, 60%, 70%, 80%, 90%, or 95% or more) of the affinity and / or specificity of the parent antibody. Typically, such functional variants retain significant sequence identity with respect to the parent sequence. Exemplary variants include variants that differ from their respective parent VH or VL regions by mutations in 12 or fewer amino acid residues, e.g., 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue, e.g., substitution, insertion, or deletion. Exemplary variants include variants that differ from the VH and / or VL and / or CDR regions of the parent sequence mainly by conservative amino acid substitutions; for example, 12 of the amino acid substitutions in the variant, e.g., 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 may be conservative. In some cases, antibodies containing variants of antibody C's VH and / or VL may be associated with higher affinity and / or specificity than the parent antibody. For the purposes of the present invention, VH and / or VL variants that enable the retention or improvement of the antibody's affinity and specificity in its binding to CD38 are particularly preferred.

[0321] For example, WO 2011 / 154453 A1 discloses CD38 antibodies containing appropriate variant CDR, VH, and VL region amino acid sequences; where amino acid residues at specific positions differ from those in the CDR, VH, and VL of antibody C shown in Table 4. These positions thus represent candidate sites where mutations in the CDR, VH, and VL sequences can be made while maintaining or improving the antibody's affinity and specificity in its binding to CD38. In particular, the positions in the VH and VL CDR that can be mutated in the functional variants of antibody C are shown in SEQ ID NO: 40-43.

[0322] Therefore, in some embodiments, one or more specific mutations occur in the CDRs described in SEQ ID NO:40-43, i.e., any functional variant of the VH and / or VL regions includes a mutation in one or more of the CDRs described in SEQ ID NO:40(VH CDR1), SEQ ID NO:41(VH CDR2), SEQ ID NO:42(VH CDR3), and SEQ ID NO:44(VL CDR3). The VH and VL regions of such antibody variants may optionally retain the original framework regions of antibody C. In one particular embodiment, the antigen-binding region includes the CDRs described in SEQ ID NO:40(VH CDR1) where X1 is S, SEQ ID NO:41(VH CDR2) where X1 is R, X2 is K, and X3 is A, SEQ ID NO:42(VH CDR3) where X1 is A, X2 is D, and X3 is V, SEQ ID NO:43(VL CDR1), AAS(VL CDR2), and SEQ ID NO:44(VL CDR3) where X1 is S. In one particular embodiment, the antigen-binding region includes the CDRs described in SEQ ID NO:40(VH CDR1) where X1 is R, SEQ ID NO:41(VH CDR2) where X1 is V, X2 is K, and X3 is T, SEQ ID NO:42(VH CDR3) where X1 is T, X2 is A, and X3 is F, SEQ ID NO:43(VL CDR1), AAS(VL CDR2), and SEQ ID NO:44(VL CDR3) where X1 is N. In one particular embodiment, the antigen-binding region includes the CDRs described in SEQ ID NO:40(VH CDR1) where X1 is S, SEQ ID NO:41(VH CDR2) where X1 is R, X2 is K, and X3 is T, SEQ ID NO:42(VH CDR3) where X1 is A, X2 is D, and X3 is V, SEQ ID NO:43(VL CDR1), AAS(VL CDR2), and SEQ ID NO:44(VL CDR3) where X1 is S.In one particular embodiment, the antigen-binding region includes the CDRs described in SEQ ID NO:40(VH CDR1) where X1 is R, SEQ ID NO:41(VH CDR2) where X1 is V, X2 is K, and X3 is V, SEQ ID NO:42(VH CDR3) where X1 is T, X2 is A, and X3 is F, SEQ ID NO:43(VL CDR1), AAS(VL CDR2), and SEQ ID NO:44(VL CDR3) where X1 is N.

[0323] In some embodiments, no mutation occurs in the CDR; that is, the functional variants in the VH and / or VL regions each retain the CDR sequences described in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:6, AAS, SEQ ID NO:7, which represent the VH CDR1-3 or VL CDR1-3 sequences of antibody C, respectively.

[0324] In one embodiment, the VH region includes an amino acid sequence having at least 80% identity with SEQ ID NO:1, or, for example, 90%, 95%, 97%, 98%, or 99% identity with SEQ ID NO:1. For example, VH may differ from SEQ ID NO:1 by mutations of 12 or fewer amino acid residues, for example, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue, for example, substitution, insertion, or deletion. In one embodiment, the VH region differs from SEQ ID NO:1 by only 12 or fewer amino acid substitutions, for example, 5 or fewer, for example, 5, 4, 3, 2, or 1 amino acid substitution. The amino acid substitutions may be, for example, conserved amino acid substitutions as described elsewhere in this specification. In certain embodiments, no mutations occur in the VH CDR, i.e., all variant VHs retain the C CDR sequences described in SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4.

[0325] In one embodiment, the VL region includes an amino acid sequence having at least 80% identity with SEQ ID NO:5, or, for example, 90%, 95%, 97%, 98%, or 99% identity with SEQ ID NO:5. For example, VL may differ from SEQ ID NO:5 by mutations of 12 or fewer amino acid residues, for example, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue, for example, substitution, insertion, or deletion. In one embodiment, the VL region differs from SEQ ID NO:5 by only 12 or fewer amino acid substitutions, for example, 5 or fewer, for example, 5, 4, 3, 2, or 1 amino acid substitution. The amino acid substitutions may be, for example, conserved amino acid substitutions as described elsewhere in this specification. In certain embodiments, no mutations occur in the VL CDR, i.e., all variant VHs retain the C CDR sequences described in SEQ ID NO:6, AAS, and SEQ ID NO:7.

[0326] In one embodiment, the antibody includes a VH region containing the sequence of SEQ ID NO:1 and a VL region containing the sequence of SEQ ID NO:5.

[0327] Fc region and CH region Mutations in amino acid residues at positions corresponding to E430, E345, and S440 (amino acid residues are numbered according to the EU index) in the human IgG1 heavy chain can enhance the ability of antibodies to induce CDC (see, for example, Example 3). While not theoretically bound, it is thought that substituting one or more amino acids at these positions can stimulate antibody oligomerization, thereby modulating effector function to enhance other relevant functions that may result in, for example, C1q binding, complement activation, CDC, ADCP, internalization, or in vivo efficacy.

[0328] The method and use of the present invention relate to an antibody comprising an antigen-binding region and an Fc region containing any of the above mutations.

[0329] In certain embodiments, an antibody that binds to human CD38 by any of the methods and uses described herein is (a) A heavy chain comprising a VH region including VH CDR1 having the sequence described in SEQ ID NO:2, VH CDR2 having the sequence described in SEQ ID NO:3, and VH CDR3 having the sequence described in SEQ ID NO:4, and a human IgG1 CH region having mutations in one or more of E430, E345, and S440 (amino acid residues are numbered according to the EU index); and (b) A light chain comprising a VL region including VL CDR1 having the sequence described in SEQ ID NO:6, VL CDR2 having the sequence AAS, and VL CDR3 having the sequence described in SEQ ID NO:7. Includes.

[0330] In other specific embodiments, an antibody that binds to human CD38 by any of the methods and uses described herein is (a) A heavy chain comprising a VH region containing SEQ ID NO:1 and a human IgG1 CH region having mutations in one or more of E430, E345 and S440 (amino acid residues are numbered according to the EU index); and (b) Light chain containing the VL region including SEQ ID NO: 5 Includes.

[0331] The antibody of the present invention comprises an Fc region or a human IgG1 CH region containing mutations in one or more of E430, E345, and S440. In the following, references to mutations in the Fc region may also apply to mutations in the human IgG1 CH region.

[0332] As described herein, the amino acid positions to be mutated in the Fc region may be provided in relation to (i.e., "correspondingly") their positions in the naturally occurring (wild-type) human IgG1 heavy chain, when numbered according to the EU index. Therefore, if the parent Fc region already contains one or more mutations, and / or if the parent Fc region is, for example, an IgG2, IgG3, or IgG4 Fc region, the amino acid position corresponding to the amino acid residue, e.g., E430, in the human IgG1 heavy chain numbered according to the EU index can be determined by alignment. Specifically, the parent Fc region is aligned with the wild-type human IgG1 heavy chain sequence to identify the residue at the position corresponding to E430 in the human IgG1 heavy chain sequence. Any wild-type human IgG1 constant region amino acid sequence containing any one of the various human IgG1 allotypes listed in Table 4 may be useful for this purpose. This is illustrated in Figure 1, which shows the alignment between two different human IgG1 allotypes—IgG1m(f) and IgG1m(a)—and wild-type human IgG2, IgG3, and IgG4, in particular the alignment of the segment corresponding to residues P247 to K447 in the human IgG1 heavy chain, where the amino acid residues are numbered according to the EU index.

[0333] Therefore, in the remaining paragraphs of this section and elsewhere in this specification, unless otherwise specified or inconsistent with the context, the amino acid positions mentioned correspond to amino acid residues in the wild-type human IgG heavy chain, and the amino acid residues are numbered according to the EU index.

[0334] In distinct and specific embodiments, the Fc region and / or human IgG1 CH region of the present invention contains mutations in only one of E430, E345, and S440; in both E430 and E345; in both E430 and S440; in both E345 and S440; or in all of E430, E345, and S440. In some embodiments, the Fc region and / or human IgG1 CH region of the present invention contains mutations in only one of E430, E345, and S440; in both E430 and E345; in both E430 and S440; in both E345 and S440; or in all of E430, E345, and S440, provided that any mutation in S440 is either S440W or S440Y. In other distinct and specific embodiments, the mutation is an amino acid substitution. In one embodiment, the mutation is an amino acid substitution in only one of E430X, E345X, and S440X; in both E430X and E345X; in both E430X and S440X; in both E345X and S440X; or in all of E430X, E345X, and S440X, wherein any mutation in S440X is preferably S440Y or S440W. More preferably, the E430X, E345X, and S440X mutations are separately selected from E430G, E345K, E430S, E430F, E430T, E345Q, E345R, E345Y, S440Y, and S440W.

[0335] In one embodiment, the mutation in one or more amino acid residues is selected from the group consisting of E430G, E345K, E430S, E430F, E430T, E345Q, E345R, E345Y, S440Y, and S440W.

[0336] In a preferred embodiment, the mutations in one or more amino acid residues are selected from the group corresponding to E430G, E345K, E430S, and E345Q.

[0337] In one embodiment, the mutation is an amino acid substitution E430X, selected from those corresponding to the amino acid residue E430, such as E430G, E430S, E430F, or E430T. In one preferred embodiment, the mutation in one or more amino acid residues includes E430G. In another preferred embodiment, the mutation in one or more amino acid residues includes E430S, and optionally, no mutation occurs in the amino acid residues corresponding to E345 and S440.

[0338] In certain preferred embodiments, the mutations in one or more amino acid residues consist of E430G, i.e., no mutations occur in the amino acid residues corresponding to E345 and S440.

[0339] In one embodiment, the mutation is an amino acid substitution E345X, selected from those corresponding to the amino acid residue E345, e.g., E345K, E345Q, E345R, and E345Y. In one preferred embodiment, the mutation in one or more amino acid residues includes E345K. In another preferred embodiment, the mutation in one or more amino acid residues includes E345Q, and optionally, no mutation occurs in the amino acid residues corresponding to E430 and S440. In a particular preferred embodiment, the mutation in one or more amino acid residues consists of E345K, i.e., no mutation occurs in the amino acid residues corresponding to E430 and S440.

[0340] In one embodiment, the mutation is an amino acid substitution S440X, typically selected from the amino acid residues corresponding to S440, such as S440Y and S440W. In one preferred embodiment, the mutation in one or more amino acid residues includes S440W, and optionally, no mutation occurs in the amino acid residues corresponding to E430 and E345. In one preferred embodiment, the mutation in one or more amino acid residues includes S440Y, and optionally, no mutation occurs in the amino acid residues corresponding to E430 and E345.

[0341] Preferably, the antibody comprises an Fc region according to any one of the sections described above, and this Fc region is a variant of a human IgG Fc region selected from the group consisting of human IgG1, IgG2, IgG3, and IgG4 Fc regions. That is, mutations in one or more amino acid residues corresponding to E430, E345, and S440 occur in the parent Fc region, which is a human IgG Fc region selected from the group consisting of IgG1, IgG2, IgG3, and IgG4 Fc regions. Preferably, the parent Fc region is a naturally occurring (wild-type) human IgG Fc region, e.g., a human wild-type IgG1, IgG2, IgG3, or IgG4 Fc region, or a mixed isotype thereof. Thus, the Fc region of the present invention may be a human IgG1, IgG2, IgG3, or IgG4 isotype, or a mixed isotype thereof, except for the described mutations (in one or more amino acid residues selected from the group corresponding to E430, E345, and S440).

[0342] In one embodiment, the parent Fc region and / or the human IgG1 CH region is the wild-type human IgG1 isotype.

[0343] Therefore, the Fc region of the present invention may be a human IgG1 Fc region, except for the described mutations (in one or more amino acid residues selected from the group corresponding to E430, E345, and S440).

[0344] In certain embodiments, the parent Fc region and / or the human IgG1 CH region are human wild-type IgG1m(f) isotypes.

[0345] In certain embodiments, the parent Fc region and / or the human IgG1 CH region are human wild-type IgG1 m(z) isotypes.

[0346] In certain embodiments, the parent Fc region and / or the human IgG1 CH region are human wild-type IgG1 m(a) isotypes.

[0347] In certain embodiments, the parent Fc region and / or the human IgG1 CH region are human wild-type IgG1 m(x) isotypes.

[0348] In certain embodiments, the parent Fc region and / or human IgG1 CH region are mixed allotype human wild-type IgG1, such as IgG1m(za), IgG1m(zax), or IgG1m(fa).

[0349] Therefore, the Fc region and / or human IgG1 CH region of the present invention may be human IgG1m(f), IgG1m(a), IgG1m(x), IgG1m(z) allotypes, or any two or more mixed allotypes thereof, except for the described mutations (in one or more amino acid residues selected from the group corresponding to E430, E345, and S440).

[0350] In certain embodiments, the parental Fc region and / or the human IgG1 CH region are human wild-type IgG1m(za) isotypes.

[0351] In certain embodiments, the parent Fc region is the human wild-type IgG2 isotype.

[0352] In certain embodiments, the parent Fc region is the human wild-type IgG3 isotype.

[0353] In certain embodiments, the parent Fc region is the human wild-type IgG4 isotype.

[0354] Table 4 shows the CH region amino acid sequences of specific examples of wild-type human IgG isotypes and IgG1 allotypes. In some embodiments, the parent Fc region contains CH2-CH3 or optionally, the hinge-CH2-CH3 segment of such a wild-type CH region amino acid sequence.

[0355] Therefore, in certain embodiments, the parent Fc region is a human wild-type IgG1 isotype containing amino acid residues corresponding to 231-447 in the human IgG1 heavy chain according to EU numbering. For example, the parent Fc region may contain amino acid residues 114-330 (directly numbered) of a sequence selected from the group consisting of SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, and SEQ ID NO:23. In certain embodiments, the parent Fc region is a human wild-type IgG1 isotype containing amino acid residues corresponding to 216-447 in the human IgG1 heavy chain according to EU numbering. For example, the parent Fc region may contain amino acid residues 99-330 (directly numbered) of a sequence selected from the group consisting of SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, and SEQ ID NO:23. As described elsewhere in this specification regarding the production of therapeutic antibodies, the C-terminal amino acid K447 may often be deleted or removed. Therefore, the parent Fc region may contain amino acid residues 114-329 (direct numbering) or amino acid residues 99-329 (direct numbering) of SEQ ID NO:45.

[0356] In certain embodiments, the Fc region of the present invention is a variant of the human wild-type IgG1 isotype containing amino acid residues corresponding to 231-447 in the human IgG1 heavy chain according to EU numbering. For example, the Fc region may contain amino acid residues 114-330 (directly numbered) of a sequence selected from the group consisting of SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33. In another embodiment, the Fc region may contain amino acid residues 114-329 (directly numbered) of SEQ ID NO:46.

[0357] In certain embodiments, the Fc region of the present invention is a variant of the human wild-type IgG1 isotype containing amino acid residues corresponding to 216-447 in the human IgG1 heavy chain according to EU numbering. For example, the Fc region may contain amino acid residues 99-330 (directly numbered) of a sequence selected from the group consisting of SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33. In another embodiment, the Fc region may contain amino acid residues 99-329 (directly numbered) of SEQ ID NO:46.

[0358] Therefore, the present invention can be applied to antibody molecules having a human IgG1 heavy chain, for example, a human IgG1 CH region amino acid sequence including SEQ ID NO:19 (IgGm(za)). Thus, the human IgG1 CH region may include the sequence of SEQ ID NO:19, excluding the described mutations.

[0359] The present invention can also be applied to antibody molecules having a human IgG1 heavy chain, for example, a human IgG1 CH region amino acid sequence including SEQ ID NO:20(IgGm(f)) or SEQ ID NO:45. Thus, the human IgG1 CH region may include the sequence of SEQ ID NO:20, excluding the described mutations. In another embodiment, the human IgG1 CH region may include the sequence of SEQ ID NO:45, excluding the described mutations.

[0360] The present invention can also be applied to antibody molecules having a human IgG1 heavy chain, for example, a human IgG1 CH region amino acid sequence including SEQ ID NO:21(IgGm(z)). Therefore, the human IgG1 CH region may include the sequence of SEQ ID NO:21, excluding the described mutations.

[0361] The present invention can also be applied to antibody molecules having a human IgG1 heavy chain, for example, a human IgG1 CH region amino acid sequence including SEQ ID NO:22 (IgGm(a)). Therefore, the human IgG1 CH region may include the sequence of SEQ ID NO:22, excluding the described mutations.

[0362] The present invention can also be applied to antibody molecules having a human IgG1 heavy chain, for example, a human IgG1 CH region amino acid sequence including SEQ ID NO:23(IgG1m(x)). Therefore, the human IgG1 CH region may include the sequence of SEQ ID NO:23, excluding the described mutations.

[0363] In other distinct and specific embodiments, the human IgG1 CH region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:24 to SEQ ID NO:33 and SEQ ID NO:45.

[0364] In certain embodiments, the human IgG1 CH region includes SEQ ID NO:24 (IgG1m(f)-E430G) or SEQ ID NO:46, and optionally, the light chain includes CL containing SEQ ID NO:37.

[0365] In certain embodiments, the antibody of the present invention is a monospecific antibody comprising two HCs and two LCs having identical amino acid sequences.

[0366] The present invention can also be applied to antibody molecules having a human IgG double chain, for example, a human IgG double chain containing the amino acid sequence of the human IgG2 CH region including SEQ ID NO:34.

[0367] The present invention can also be applied to antibody molecules having a human IgG triple chain, for example, a human IgG triple chain containing the human IgG3 CH region amino acid sequence including SEQ ID NO:35.

[0368] The present invention can also be applied to antibody molecules having a human IgG quadrichain, for example, a human IgG quadrichain containing the human IgG4 CH region amino acid sequence including SEQ ID NO:36.

[0369] However, Fc regions containing one or more further mutations, i.e., mutations in one or more other amino acid residues other than those corresponding to E430, E345, and S440 in the human IgG monohelic acid when numbered according to the EU index, are also intended in the antibodies disclosed herein. Furthermore or alternatively, the Fc region may be a mixed isotype, for example, in this case different CH regions originate from different IgG isotypes. Thus, as will be described in more detail below, the parental Fc region may already contain one or more further mutations compared to such a wild-type (naturally occurring) human IgG Fc region, or it may be a mixed isotype.

[0370] In one embodiment, a parent Fc region into which a mutation has been introduced in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 is, for example, a human IgG Fc region containing one or more further mutations compared to the wild-type human IgG1, IgG2, IgG3, and IgG4 Fc regions described in one of SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:34, SEQ ID NO:35, and SEQ ID NO:36. Expressed in another way, an Fc region containing mutations at E430, E345 and / or S440 may also differ from a reference Fc region, e.g., a reference wild-type human IgG1, IgG2, IgG3 and IgG4 Fc region, as described in, for example, one of SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36, by one or more further mutations. For example, an Fc region may differ from a wild-type Fc region by mutations of 12 or fewer amino acid residues, e.g., 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid residue, e.g., substitution, insertion or deletion. For example, the (Eu-numbered) C-terminal amino acid Lys(K) at position 447 may be deleted. Some host cells used for antibody production may contain enzymes that can remove Lys at position 447, and such removal may not be homogeneous. Therefore, to increase the homogeneity of the product, therapeutic antibodies may be produced without C-terminal Lys(K). Methods for producing antibodies without C-terminal Lys(K) are well known to those skilled in the art and include genetic manipulation of nucleic acids expressing the antibody, enzymatic methods, and the use of specific host cells. Therefore, for example, the parent Fc region may include the sequence described in SEQ ID NO:45.

[0371] Preferably, any one or more such further mutations do not reduce the ability of the antibody disclosed herein, i.e., the antibody containing a mutation in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 in the human IgG1 heavy chain, to induce CDC and / or ADCC. More preferably, any one or more such further mutations do not reduce the ability of the antibody to induce CDC. Most preferably, any one or more such further mutations do not reduce the ability of the antibody to induce either CDC or ADCC. Candidates for one or more further mutations can be tested in CDC or ADCC assays, for example, as disclosed herein, e.g., Examples 3 and 4. For example, the CDC of the antibody described herein, e.g., IgG1-C-E430G, can be tested with and without a specific candidate for one or more further mutations in the assay of Example 3 or the assays described in the following sections (or similar assays) to determine the effect of candidate further mutations on the antibody's ability to induce CDC. Similarly, the ADCC of the antibodies described herein, for example IgG1-C-E430G, can be tested with and without specific candidate further mutations in the assay of Example 4 or the assays described in the following sections (or similar assays) to determine the effect of further candidate mutations on the antibody's ability to induce ADCC.

[0372] Preferably, in the antibody of the present invention comprising two HCs and two LCs, the Fc regions in the first and second HCs are identical, and as a result, the Fc region is a homodimer in a dimerized form.

[0373] However, in some embodiments, in an antibody comprising two HCs and two LCs, the Fc region of the first HC may differ from the Fc region of the second HC by one or more amino acids, and as a result, the Fc region is heterodimer in a dimerized form. For example, a mutation in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 (amino acid residues are numbered according to the EU index) in the IgG1 heavy chain may be present in only one of the Fc regions. Therefore, in some embodiments, one Fc region may be SEQ ID NO:45 or a human wild-type IgG Fc region selected from SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36, and the other Fc region may be identical except for a mutation in one or more amino acid residues selected from the group corresponding to E430, E345 and S440 in the IgG1 heavy chain.

[0374] In one embodiment, the antibodies according to any aspect or embodiment of this specification are human antibodies, except for the described mutations.

[0375] In one embodiment, the antibody according to any aspect or embodiment of this specification is a full-length antibody, such as a human full-length antibody, with the exception of the described mutations.

[0376] In one embodiment, the antibody according to any aspect or embodiment of this specification is a bivalent antibody, such as a human bivalent antibody, such as a human bivalent full-length antibody, with the exception of the described mutations.

[0377] In one embodiment, the antibody according to any aspect or embodiment of this specification is a monoclonal antibody, such as a human monoclonal antibody, such as a human bivalent monoclonal antibody, such as a human bivalent full-length monoclonal antibody, with the exception of the described mutations.

[0378] In preferred embodiments, the antibody according to any aspect or embodiment of this specification is, with the exception of the described mutations, an IgG1 antibody, e.g., a full-length IgG1 antibody, e.g., a human full-length IgG1 antibody, and optionally, a human monoclonal full-length bivalent IgG1,κ antibody, e.g., a human monoclonal full-length bivalent IgG1m(f),κ antibody.

[0379] The antibodies according to the present invention are advantageously in a bivalent, monospecific format, comprising two antigen-binding regions that bind to the same epitope. However, a bispecific format in which one of the antigen-binding regions binds to a different epitope is also intended. Thus, antibodies according to any aspect or embodiment of this specification may be either monospecific or bispecific antibodies, as long as it is not inconsistent with the context.

[0380] Therefore, in one embodiment, the antibody according to any aspect or embodiment of this specification is a monospecific antibody, e.g., a human monospecific antibody, e.g., a full-length human monospecific antibody, e.g., a full-length human monospecific bivalent monoclonal antibody, e.g., a full-length human bivalent monospecific monoclonal antibody, e.g., a full-length human bivalent monospecific antibody, except for the described mutations.

[0381] In another aspect, the antibody according to any aspect or aspect of this specification is a bispecific antibody, e.g., a full-length bispecific antibody, and optionally, a full-length bispecific divalent IgG1,κ antibody, except for the mutations described herein.

[0382] formulation In one aspect, the antibodies of the present invention are included in pharmaceutical compositions such as those described in WO 2021 / 144457 A1.

[0383] Therefore, in one aspect, the antibody of the present invention is included in a pharmaceutical composition, and the pharmaceutical composition is (a) Antibodies; (b) 5-40 mM histidine or acetic acid; (c) 100-400 mM sorbitol or sucrose; and (d) Surfactants It includes, consists of, or essentially consists of.

[0384] In one aspect, the antibody of the present invention is contained in a pharmaceutical composition, and the pharmaceutical composition is in an aqueous solution state. a) Antibodies; b) 5-40 mM histidine or acetic acid; c) 100-400 mM sorbitol or sucrose; and d) Surfactants It consists of.

[0385] In some embodiments, the pharmaceutical composition contains a) 1-80 mg / mL, e.g., 1-60 mg / mL, 1-40 mg / mL, 1-30 mg / mL, or 1-25 mg / mL; 2-80 mg / mL, e.g., 2-40 mg / mL, or 2-30 mg / mL; or 10-80 mg / mL, e.g., 10-40 mg / mL, or 10-30 mg / mL; or 15-80 mg / mL, e.g., 15-40 mg / mL, e.g., 15-25 mg / mL, e.g., 2 mg / mL, 4 mg / mL, 6 mg / mL, 8 mg / mL, 10 mg / mL, 12 mg / mL, 14 mg / mL, 16 mg / mL, 18 mg / mL, 20 mg / mL, 22 mg / mL, 24 mg / mL, 26 mg / mL, 28 mg / mL, 30 mg / mL, 32 mg / mL, 34 mg / mL, 36 mg / mL, 38 mg / mL The antibody may be mg / mL, 40 mg / mL, or 50 mg / mL. In one embodiment, a) is an antibody of approximately 20 mg / mL. In a particularly intended embodiment, a) is an antibody of 20 mg / mL.

[0386] In some embodiments, b) in the pharmaceutical composition may be histidine or acetic acid in a concentration of 5–30 mM, e.g., 5–25 mM, e.g., 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, or 30 mM. In one embodiment, b) is histidine or acetic acid in a concentration of about 20 mM, e.g., 20 mM. In one particular embodiment, b) is acetic acid. In an embodiment particularly intended, b) is histidine.

[0387] In some embodiments, c) in the pharmaceutical composition may be sorbitol or sucrose in an amount of 100-350 mM, e.g., 100-300 mM, 100-260 mM, 100-200 mM, 150-350 mM, 200-300 mM, 200-260 mM, 200-350 mM, 200-300 mM, 200-260 mM, 230-350 mM, 230-300 mM, 230-260 mM, or 240-260 mM; e.g., 245 mM, 246 mM, 247 mM, 248 mM, 249 mM, 250 mM, 251 mM, 252 mM, 253 mM, 254 mM, or 255 mM. In one embodiment, c) is sorbitol or sucrose in an amount of about 250 mM, e.g., 250 mM. In one embodiment, c) is sucrose. In a particularly intended embodiment, c) is sorbitol.

[0388] The pharmaceutical composition may have a pH of, for example, 5.0-6.5, 5.5-6.5, 5.6-6.5, 5.7-6.5, 5.8-6.5, 5.9-6.5, 6.0-6.5, 5.5-6.4, 5.5-6.3, 5.5-6.2, 5.5-6.1, 5.5-6.0, 5.7-6.3, 5.8-6.2, or 5.9-6.1. In one embodiment, the pH is approximately 6. In one embodiment, the pH is 6, for example, 6.0.

[0389] Surfactants suitable for pharmaceutical compositions are known in the art and can be selected from the group including, for example, glycerol monooleate, benzethonium chloride, sodium doxate, phospholipids, polyethylene alkyl ethers, sodium lauryl sulfate and tricaprylin, benzalkonium chloride, citrimide, cetylpyridinium chloride and phospholipids, α-tocopherol, glycerol monooleate, myristyl alcohol, phospholipids, poloxamer, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, polyoxyl hydroxystearate, polyoxylglycerides, polysorbates, propylene glycol dilaurate, propylene glycol monolaurate, sorbitan ester sucrose palmitate, sucrose stearate, tricaprylin, and TPGS. In one particular embodiment, the surfactant is polysorbate. Preferably, the surfactant is polysorbate 20 or 80. In one embodiment, the surfactant is polysorbate 20 (PS20). In a particularly intended embodiment, the surfactant is polysorbate 80 (PS80).

[0390] The surfactant concentration is typically about 0.005% to 0.5% w / v, for example about 0.01 to 0.1% w / v, for example about 0.01 to 0.09% w / v, for example about 0.01 to 0.06% w / v, for example about 0.01 to 0.05% w / v, for example 0.02% w / v, or 0.03% w / v, or 0.04% w / v, or 0.05% w / v, or 0.06% w / v. In one embodiment, the surfactant concentration is about 0.04% w / v, for example 0.04% w / v.

[0391] In one embodiment, the pharmaceutical composition has a pH of 5.9 to 6.1, for example, a pH of about 6 or 6.0, and a) Antibody at 1-80 mg / mL, b) 15-40 mM histidine c) 200-300 mM sorbitol, d) 0.01%~0.1% w / v surfactant It includes, or essentially consists of, them.

[0392] In one embodiment, the pharmaceutical composition has a pH of 5.9 to 6.1, for example, a pH of about 6 or 6.0, and a) 10-40 mg / mL antibody, b) 15-40 mM histidine c) 200-300 mM sorbitol, d) 0.02%~0.06% w / v surfactant It includes, or essentially consists of, them.

[0393] In one embodiment, the pharmaceutical composition has a pH of 5.9 to 6.1, for example, a pH of about 6 or 6.0, and a) 10-40 mg / mL antibody, b) Histidine at 15-25 mM c) 240-260 mM sorbitol, d) 0.02%~0.06% w / v surfactant It includes, or essentially consists of, them.

[0394] In certain embodiments, the surfactant in d) is a polysorbate, for example, polysorbate 20 or polysorbate 80. In one embodiment, the surfactant is polysorbate 20. In one particularly intended embodiment, the surfactant is polysorbate 80.

[0395] As one specific example, the pharmaceutical composition has a pH of about 6, and a) Approximately 20 mg / mL of antibody, b) Histidine at approximately 20 mM, c) Approximately 250 mM sorbitol, and d) Polysorbate 80 at approximately 0.04% w / v It includes, or essentially consists of, them.

[0396] As another specific example, a pharmaceutical composition having a pH of 6 and in aqueous solution form, a) 20 mg / mL antibody, b) 20 mM histidine, c) 250 mM sorbitol, and d) 0.04% w / v polysorbate 80 It includes, consists of, or essentially consists of.

[0397] In some embodiments, the pharmaceutical compositions according to the present invention are typically concentrates that are diluted before or in connection with administration to a subject or patient. Suitable diluents are known in the art. Preferred diluents include, but are not limited to, physiological saline (0.9% NaCl) and dextrose in aqueous solution (e.g., 5% w / v).

[0398] A pharmaceutical composition according to any aspect or aspect of this specification may contain a CD38 antibody further characterized by other or additional features as described elsewhere in this specification.

[0399] Therefore, in certain embodiments, the pharmaceutical composition has a pH of about 6, a) An antibody that binds to human CD38 at a concentration of approximately 20 mg / mL, b) Histidine at approximately 20 mM, c) Approximately 250 mM sorbitol, and d) Polysorbate 80 at approximately 0.04% w / v including or essentially derived from them The antibody is a full-length bivalent antibody containing, consisting of, or essentially consisting of, two heavy chains and two light chains. Each heavy chain includes a VH region and a CH region, the VH region includes SEQ ID NO:1, and the CH region includes SEQ ID NO:24 or SEQ ID NO:46, and Each light chain includes a VL region and a CL region, the VL region including SEQ ID NO: 5 and the CL region including SEQ ID NO: 37.

[0400] In one embodiment, the pharmaceutical composition has a pH of about 6 and is in an aqueous solution state. a) 20 mg / mL antibody, b) 20 mM histidine, c) 250 mM sorbitol, and d) 0.04% w / v polysorbate 80 It includes, consists of, or essentially consists of.

[0401] use In one aspect, the present invention relates to an anti-CD38 antibody or a (pharmaceutical) composition comprising the antibody, in any aspect or embodiment of the present invention, for use in treating or preventing hematological malignancies involving CD38-expressing cells, such as in the treatment or prevention of (relapsed or refractory) multiple myeloma, for example, the antibody is administered to the subject in a dose of at least (about) 4 mg / kg body weight, for example, 4 mg / kg to (about) 24 mg / kg body weight or 8 mg / kg to (about) 16 mg / kg body weight, preferably 16 mg / kg body weight.

[0402] In one aspect, the present invention relates to an anti-CD38 antibody or a (pharmaceutical) composition containing the antibody, in any aspect or embodiment of the present invention, for use in treating or preventing hematological malignancies in subjects containing cells expressing human CD38, such as those described herein, for use in the treatment or prevention of (relapsed or refractory) multiple myeloma, for example, the antibody is administered to the subject in a dose of at least (about) 4 mg / kg body weight, for example, 4 mg / kg to (about) 24 mg / kg body weight or 8 mg / kg to (about) 16 mg / kg body weight, preferably 16 mg / kg body weight.

[0403] In one aspect, the present invention provides an anti-CD38 antibody or a (pharmaceutical) composition comprising the antibody in any of the embodiments and aspects described herein for use in the prevention or treatment of hematological malignancies, for example, for use in the treatment of (relapsed or refractory) multiple myeloma, for example, the antibody is administered to the subject in a dose of at least (about) 4 mg / kg body weight, for example, 4 mg / kg to (about) 24 mg / kg body weight or 8 mg / kg to (about) 16 mg / kg body weight, preferably 16 mg / kg body weight.

[0404] In one aspect, the present invention relates to an anti-CD38 antibody or a (pharmaceutical) composition comprising the antibody, as described herein, for pharmaceutical use for the treatment or prevention of hematological malignancies, for example, for pharmaceutical use for the treatment or prevention of (relapsed or refractory) multiple myeloma, for example, the antibody is administered to the subject in a dose of at least (about) 4 mg / kg body weight, for example, 4 mg / kg to (about) 24 mg / kg body weight or 8 mg / kg to (about) 16 mg / kg body weight, preferably 16 mg / kg body weight.

[0405] In one aspect, the present invention provides the use of an anti-CD38 antibody or a (pharmaceutical) composition containing the antibody, in any of the embodiments and aspects described herein, for the manufacture of a pharmaceutical for the prevention or treatment of hematological malignancies, for example, for the manufacture of a pharmaceutical for the treatment or prevention of (relapsed or refractory) multiple myeloma, for example, the antibody is administered to the subject in a dose of at least (about) 4 mg / kg body weight, for example, 4 mg / kg to (about) 24 mg / kg body weight or 8 mg / kg to (about) 16 mg / kg body weight, preferably 16 mg / kg body weight.

[0406] array (Table 4) Amino acid sequences and nucleic acid sequences TIFF2026513656000006.tif156167TIFF2026513656000007.tif224167TIFF2026513656000008.tif229167 TIFF2026513656000009.tif229167TIFF2026513656000010.tif229167TIFF2026513656000011.tif250167

[0407] List of References Each document cited in this list, or elsewhere in this specification, is incorporated herein by reference in its entirety. TIFF2026513656000012.tif187167TIFF2026513656000013.tif222167TIFF20265136560 00014.tif222167TIFF2026513656000015.tif209167TIFF2026513656000016.tif146166 [Examples]

[0408] The present invention is further illustrated by the following embodiments, which should not be construed as limiting.

[0409] Example 1 - Antibody Production and Formulation Antibody expression construct The antibodies were essentially prepared as described in WO 2020 / 012036 A1, WO 2020 / 012038 A1, and WO 2021 / 144457 A1 (all incorporated herein by reference).

[0410] In short, for the expression of human antibodies and humanized antibodies used herein, variable heavy chain (VH) and variable light chain (VL) sequences were prepared by gene synthesis (GeneArt gene synthesis; ThermoFisher Scientific) and cloned into a pcDNA3.3 expression vector (ThermoFisher Scientific) containing the constant region of the human IgG heavy chain (HC) (constant region human IgG1m(f)HC: SEQ ID NO: 20) and / or the constant region of the human κ light chain (LC): SEQ ID NO: 37. The desired mutations were introduced by gene synthesis. The CD38 antibody variants described herein have VH and VL sequences derived from previously described CD38 antibodies IgG1-A (WO 2006 / 099875 A1, WO 2008 / 037257 A2, WO 2011 / 154453 A1; VH:SEQ ID NO:10; VL:SEQ ID NO:11), IgG1-B (WO 2006 / 099875 A1, WO 2008 / 037257 A2, WO 2011 / 154453 A1; VH:SEQ ID NO:8; VL:SEQ ID NO:9), and IgG1-C (WO 2011 / 154453 A1; VH:SEQ ID NO:1; VL:SEQ ID NO:5). In some experiments, human IgG1 antibody b12, an HIV gp120-specific antibody, was used as a negative control (Barbas et al., J Mol Biol. 1993 Apr 5;230(3):812-23;VH:SEQ ID NO:12;VL:SEQ ID NO:16).

[0411] Transient expression antibody constructs Plasmid DNA mixtures encoding both the heavy and light chains of the antibody were transiently transfected into Expi293F cells (Gibco, catalog number A14635) using 293fectin (Life Technologies), as essentially described by Vink et al. (Vink et al., 2014 Methods 65(1):5-10). The antibody concentration in the supernatant was measured by absorbance at 280 nm. The antibody-containing supernatant was used directly in in vitro assays, or the antibody was purified as described below.

[0412] Antibody purification and quality assessment The antibody was purified by protein A affinity chromatography. The culture supernatant was filtered through a 0.20 μM dead-end filter, loaded onto a 5 mL MabSelect SuRe column (GE Healthcare), washed, and eluted with 0.02 M sodium citrate-NaOH, pH 3. Immediately after purification, the eluate was loaded onto a HiPrep Desalting column (GE Healthcare), and the antibody was buffered with 12.6 mM NaH2PO4, 140 mM NaCl, pH 7.4 buffer (B. Braun or Thermo Fisher). After buffer exchange, the sample was sterile filtered through a 0.2 μm dead-end filter. The purified protein was analyzed by several biological analysis assays, including capillary electrophoresis (CE-SDS) and high-performance size exclusion chromatography (HP-SEC) on a sodium dodecyl sulfate-polyacrylamide gel. The concentration was measured by absorbance at 280 nm. The purified antibody was stored at 2–8°C.

[0413] IgG1-C-E430G contains VH with SEQ ID NO:1, VL with SEQ ID NO:5, CH with SEQ ID NO:46, and CL with SEQ ID NO:37. IgG1-C-E430G can be expressed in CHO cells.

[0414] Antibody preparations The antibody IgG1-C-E430G used in the clinical trials described in the examples below was essentially prepared and formulated as described in WO 2021 / 144457 A1 (incorporated herein by reference). The optimal formulation of IgG1-C-E430G was found to be 20 mM histidine, 250 mM sorbitol, 0.04% (w / v) PS80, and pH 6.0. The formulation contained 20 mg / mL of antibody.

[0415] Example 2 - Clinical Trial Clinical trial design: An open-label, multicenter, phase 1 / 2 clinical trial of IgG1-C-E430G was conducted to evaluate the safety, tolerability, pharmacokinetics, pharmacodynamics, immunogenicity, and preliminary efficacy of IgG1-C-E430G in patients with RRMM and other hematological malignancies, including R / R DLBCL. The clinical trial consisted of three parts: dose escalation (phase 1), expansion part A (IgG1-C-E430G single cohort) (phase 2), and expansion part B (randomized direct comparison) (phase 2). Figure 2 shows a schematic diagram of the clinical trial design.

[0416] Dosage gradual increase The dose escalation portion was designed to evaluate IgG1-C-E430G in subjects with RRMM and determine the recommended phase 2 dose (RP2D).

[0417] IgG1-C-E430G was evaluated at six different dose levels in subjects with RRMM. The initial dose of cycle 1 was divided into two doses administered on consecutive days. Table 5 shows the doses of IgG1-C-E430G administered in subjects with RRMM.

[0418] (Table 5) Subjects with RRMM (Rapid-Reaching Measuring Drug Use) after IgG1-C-E430G dose administration (dose escalation) TIFF2026513656000017.tif173167 Note: In MM-DL1, divided doses are administered on days 1-2 and 8-9 of cycle 1; the prescribed dose is administered on subsequent administration days. In all other MM-DL cohorts, divided doses are administered only on days 1-2 of cycle 1; the prescribed dose is administered on subsequent administration days. Based on the newly generated data, additional doses, such as 20 mg / kg, or changes to the administration schedule may be considered. DL = dose level; RRMM = relapsed or refractory multiple myeloma; MM = multiple myeloma.

[0419] IgG1-C-E430G was administered to the RRMM cohort as an IV infusion in a 4-week, or 28-day, cycle, as follows: • Cycle 1: Days 1, 2, 8, 15, and 22 (Q1W). Note that the initial dose of IgG1-C-E430G was split over two consecutive days (i.e., C1D1 and C1D2) and in RRMM, MM-DL1 was administered on day 9. • Cycle 2: Days 1, 8, 15, and 22 (Q1W) • Cycles 3-6: Days 1 and 15 (Q2W) • Cycle 7 and beyond: Day 1 (Q4W) Each infusion was estimated to last between 1 and 8 hours.

[0420] Figure 2 shows a schematic diagram of the clinical trial design for IgG1-C-E430G.

[0421] Expanded Part A The objective of Expanded Part A is to provide further data on efficacy, safety, tolerability, pharmacokinetics, pharmacodynamics, and biomarkers.

[0422] Expanded Part A is designed to include subjects with RRMM and R / R DLBCL in the cohort, as follows: • The data includes the first 10 evaluable response subjects and anti-CD38 mAb naive subjects with RRMM who received IgG1-C-E430G at 16 mg / kg or 24 mg / kg from dose escalation and / or expanded Part A. • Subjects: 20 patients with CD38 mAb-refractory RRMM • Up to 40 individuals with R / R DLBCL are included.

[0423] Expanded Part B In the expanded Part B (randomized H2H) study, IgG1-C-E430G IV will be compared to a reference anti-CD38 antibody (without E340G, i.e., subcutaneous (SC) daratumumab-DARZALEX FASPRO®) in a population of anti-CD38 mAb-naive RRMM patients.

[0424] Subject Information Participants are eligible to take part in the clinical trial only if they meet all of the following criteria: Participants must be at least 18 years old. The target has RRMM. The subjects must have documented multiple myeloma as defined by the following criteria and have evidence of disease progression with their most recent previous treatment regimen, based on the IMWG criteria: • A previous record of monoclonal plasma cells in ≥10% of the bone marrow, or the presence of a plasmacytoma confirmed by biopsy; and • Measurable lesions at baseline as defined by any of the following: • IgG, IgA, IgD, or IgM myeloma: Serum M protein level ≥ 0.5 g / dL (≥ 5 g / L) or urinary M protein level ≥ 200 mg / 24 hours; or • Light chain myeloma: Serum Ig free light chain (FLC) ≥ 10 mg / dL and abnormal serum IgκλFLC ratio. The subjects must have exhausted all standard treatment options, as determined by the principal investigator.

[0425] Subjects with R / R DLBCL Participants may have either new-onset DLBCL or histologically transformed DLBCL. Participants with R / R DLBCL must have exhausted standard treatment options at the discretion of the principal investigator.

[0426] exposure The median duration of IgG1-C-E430G infusion at the prescribed dose was 3.80 hours (range: 1.15–4.57 hours).

[0427] Example 3 - Clinical Efficacy result Table 6 shows the best overall response at each dose level among 21 evaluable subjects with RRMM (data cutoff date: October 3, 2022). During the dose escalation phase of the IgG-C-E430G clinical trial, preliminary antitumor activity was observed at DL of 4 mg / kg or higher. At 16 mg / kg DL, of the 9 evaluable subjects, 3 were anti-CD38 naive, 1 had a minimal response; and 6 had been previously exposed to anti-CD38 antibodies, with 1 of each having a partial response and 1 minimal response (Tables 7 and 8).

[0428] Table 7 shows the best overall response at each dose level among five anti-CD38 mAb naive subjects for whom response was evaluable. Two subjects achieved complete response (CR; one at 4 mg / kg and one at 24 mg / kg), and one subject had minimal response (MR) at 16 mg / kg.

[0429] Table 8 shows the best overall response at each dose level among 16 subjects treated with anti-CD38 mAbs. The best observed response was a partial response (PR) in one subject at 16 mg / kg; in addition, two subjects achieved a reduced response (MR) (one at 8 mg / kg and one at 16 mg / kg).

[0430] In summary, data from the dose-escalation part of the IgG-C-E430G clinical trial indicate that IgG1-C-E430G has promising clinical activity in patients with anti-CD38 mAb naive RRMM. Patients with RRMM who have a history of anti-CD38 mAb treatment, such as those with multiple prior treatments including proteosome inhibitors, immunomodulators, and anti-CD38 mAb therapy, may also potentially benefit from IgG1-C-E430G treatment.

[0431] (Table 6) Best Overall Efficiency - All RRMMs TIFF2026513656000018.tif186167[a] Based on the Clopper and Pearson Act. [b] ORR includes patients with the best CR and PR effects; clinical benefit includes ORR and MR.

[0432] (Table 7) Best Overall Effect - Anti-CD38 mAb Naive RRMM TIFF2026513656000019.tif134167[a] Based on the Clopper and Pearson Act. [b] ORR includes patients with the best CR and PR effects; clinical benefit includes ORR and MR.

[0433] (Table 8) Best overall response - RRMM treated with anti-CD38 mAb TIFF2026513656000020.tif166167[a] Based on the Clopper and Pearson Act. [b] ORR includes patients with the best CR and PR effects; clinical benefit includes ORR and MR.

[0434] Example 4 - Clinical Safety result As of October 3, 2022, all 24 subjects (100%) in the dose-escalation group experienced at least one study-induced adverse event (TEAE); 21 subjects (87.5%) experienced a TEAE considered to be associated with IgG1-C-E430G (Tables 9 and 10). The most common TEAEs (≥20% of subjects) were IRR (18 subjects; 75.0%), neutropenia (15 subjects; 62.5%), diarrhea (10 subjects; 41.7%), anemia (10 subjects; 41.7%), COVID-19 (6 subjects; 25.0%), fever (5 subjects; 20.8%), thrombocytopenia (5 subjects; 20.8%), and blurred vision (5 subjects; 20.8%); see Table 9.

[0435] Severe TEAEs were reported in 15 subjects (62.5%) (Table 11). The most common severe TEAEs (≥5% of subjects) were COVID-19 (3 ​​subjects; 12.5%), fever (3 subjects; 12.5%), IRR (2 subjects; 8.3%), and pneumonia (2 subjects; 8.3%). Four subjects (16.7%) experienced severe TEAEs considered to be associated with IgG1-C-E430G: IRR in 2 subjects (8.3%), appendicitis, neutropenic sepsis, and fungal pneumonia in 1 subject (4.2%), and increased fibrin D-dimer in 1 subject (4.2%); see Table 12.

[0436] Eighteen subjects (75.0%) experienced an internal rate of return (IRR) at DL doses of 4, 8, 16, and 24 mg / kg (Table 6).

[0437] No cases of cytokine release syndrome were reported. There were no fatal TEAEs. All subjects tested negative for anti-drug antibodies in immunogenicity analysis.

[0438] Twenty-one subjects (87.5%) discontinued treatment in the clinical trial, while three subjects (12.5%) continued receiving IgG1-C-E430G. Reasons for discontinuation included disease progression (13 subjects; 54.2%), adverse events (AEs) (4 subjects; 16.7%), clinical progression (3 subjects; 12.5%), and requests from subjects discontinuing treatment in the clinical trial (1 subject; 4.2%). AEs leading to treatment discontinuation included internal rate of reaction (IRR) (2 subjects; 8.3%), neutropenia (1 subject; 4.2%), neutropenic sepsis (1 subject; 4.2%), polynuclear respiratory virus infection (1 subject; 4.2%), and thrombocytopenia (1 subject; 4.2%).

[0439] As of October 3, 2022, the overall data indicates that 16 mg / kg DL can deliver clinical activity while ensuring a tolerable safety profile; therefore, 16 mg / kg was selected as the RP2D for the expanded cohort in RRMM and R / R DLBCL in the IgG-C-E430G clinical trial.

[0440] Clinical safety data from the dose-escalation part of the IgG-C-E430G clinical trial, involving subjects with RRMM, demonstrated good tolerability in 11 subjects treated with IgG1-C-E430G at 16 mg / kg DL. Among the 11 subjects treated with 16 mg / kg, the most common TEAEs (≧20%) were IRR (9 subjects, 81.8%; all were associated), neutropenia (7 subjects, 63.6%; 6 were associated), diarrhea (5 subjects, 45.5%; 2 were associated), fatigue (3 subjects, 27.3%; 2 were associated), blurred vision (3 subjects, 27.3%; 1 was associated), and COVID-19 (3 ​​subjects, 27.3%; not associated); see Tables 9 and 10. At 16 mg / kg DL, serious TEAEs were reported in 6 subjects (54.5%; Table 11). One subject (9.1%) experienced one of the following serious TEAEs considered unrelated to IgG1-C-E430G: COVID-19, pneumonia, dyspnea, rhinovirus infection, acute myocardial infarction, hypervolemia, and pneumococcal sepsis. One subject (9.1%) experienced a serious TEAE and IRR considered related to IgG1-C-E430G; see Table 12. In the 16 mg / kg DL group, 10 subjects (90.9%) discontinued treatment in the clinical trial, and 1 subject (9.1%) continued receiving IgG1-C-E430G. Reasons for discontinuation included disease progression (5 subjects; 45.5%), clinical progression (3 subjects; 27.3%), and adverse events (2 subjects; 18.2%; both due to IRR). The median treatment duration was 2.5 months (range: 0.2–5.1 months). IgG1-C-E430G.

[0441] In summary, data from the dose-escalation phase of the IgG-C-E430G clinical trial suggest that IgG1-C-E430G has an acceptable safety profile with no tumor lysis syndrome or cytokine release syndrome events and no treatment-related deaths.

[0442] (Table 9) Adverse events that occurred during the study treatment TIFF2026513656000021.tif140167TIFF2026513656000022.tif227167TIFF2026513656000023.tif227167TIFF2026513656000024.tif227167TIFF2026513656000025.tif47167 Percentages are based on N, and multiple adverse events with the same base word are counted only once per subject. This is coded by MedDRA version 25.0. Adverse events that occurred during the study treatment were AEs that began after the first IMP and within 30 days of the last IMP, or pre-existing AEs that worsened.

[0443] (Table 10) Adverse events that occurred under relevant study treatments TIFF2026513656000026.tif207167 Percentages are based on N, and multiple adverse events with the same basic word are counted only once per subject. The correlation will be assessed by the principal investigator. This is coded by MedDRA version 25.0. Adverse events that occurred during the study treatment were AEs that began after the first IMP and within 30 days of the last IMP, or pre-existing AEs that worsened.

[0444] (Table 11) Serious adverse events that occurred during experimental treatment TIFF2026513656000027.tif167167 Percentages are based on N, and multiple adverse events with the same basic word are counted only once per subject. This is coded by MedDRA version 25.0. A serious adverse event occurring during a serious investigational treatment is a serious AE that began after the first IMP and within 30 days of the last IMP, or a pre-existing AE that became serious.

[0445] (Table 12) Related serious adverse events that occurred during investigational treatment TIFF2026513656000028.tif64170 Percentages are based on N, and multiple adverse events with the same basic term are counted only once per subject. Relevance is assessed by the principal investigator. This is coded by MedDRA version 25.0. Adverse events that occurred during the study treatment were defined as serious AEs that began after the first IMP and within 30 days of the last IMP, or pre-existing AEs that became serious.

[0446] (Table 13) Overview of infusion reactions by dose level TIFF2026513656000029.tif96170 Infusion reactions are limited to events marked as AESI, and the basic term is infusion reaction.

[0447] Example 5: Evaluation of lymphocyte population in whole blood of patients administered IgG1-C-E430G. method In the dose-escalation phase of the first-in-human clinical trial of IgG1-C-E430G, we evaluated the whole blood NK and T cell populations from RRMM patients who received IgG1-C-E430G at doses of 0.2 / 0.6–24 mg / kg.

[0448] As of the data cutoff date of October 3, 2022, preliminary pharmacodynamic data were available from 24 subjects with RRMM (Reduced Risk of Malnutrition) who received IgG1-C-E430G in the dose-escalation part (0.2 / 0.6~24 mg / kg) of the first-in-human clinical trial.

[0449] As of the data cutoff date of August 14, 2023, preliminary pharmacodynamic data were available from 24 subjects with RRMM (Responsible Rate of Malignancy) who received IgG1-C-E430G in the dose-escalation part (0.2 / 0.6~24 mg / kg) of the first-in-human clinical trial.

[0450] Blood samples for immunophenotyping (IPT) were collected from patients according to the scheme shown in Table 14. The blood was collected in 5 mL EDTA blood collection tubes and stored at room temperature.

[0451] (Table 14) Blood sampling schedule TIFF2026513656000030.tif67167 * Dosage level 1 (0.2 / 0.6 mg / kg only)

[0452] For each blood sample, 50 μL of whole blood was added to a polystyrene tube, and 78.75 μL of the antibody cocktail (Table 15) was added to each tube. The tubes were then gently mixed using a vortex mixer. After incubation in the dark at room temperature for 15 minutes, 450 μL of 1× FACS Lyse buffer (BD Bioscience, USA; catalog number 349202) was added, and the samples were incubated in the dark at room temperature for 15 minutes, followed by analysis using a FACSCanto flow cytometer (BD Biosciences, USA). Various lymphocyte subpopulations were identified as follows: ·NK cells: CD3- / CD56+ / CD16+ ·T cells: CD3+ • Single ball: CD14+ ·B cells: CD3- / CD19+ ·NKT cells: CD3+ / CD56+ / CD16+.

[0453] (Table 15) Components of the antibody cocktail used TIFF2026513656000031.tif74167

[0454] Results: Number of NK cells in peripheral blood of patients administered IgG1-C-E430G. Figure 3 shows that IgG1-C-E430G administration was associated with a rapid decrease in peripheral blood NK cell (CD3- / CD56+ / CD16+ cell) counts at all evaluated dose levels in all subjects. The median maximum reduction in NK cell count compared to baseline (C1D1, pre-administration) was 97% (range 66%–100%, n=21) in evaluable patients. Patients with BL values ​​below LLOQ, patients with no BL value, or patients with only a BL value were excluded from this analysis (Figure 4). NK cell counts remained low during IgG1-C-E430G treatment in the majority of patients. Table 16 summarizes the baseline, maximum change, and percentage of peripheral blood NK cell counts in patients treated with IgG1-C-E430G.

[0455] In conclusion, a significant reduction in NK cells was observed in patients administered IgG1-C-E430G at all evaluated dose levels. The observed decrease in NK cells confirms the biological activity of IgG1-C-E430G in patients and indicates its ADCC activity.

[0456] (Table 16) Peripheral blood NK cell population of patients administered IgG1-C-E430G TIFF2026513656000032.tif174159 Cutoff date: October 3, 2022

[0457] Results: Peripheral blood T cell count of patients administered IgG1-C-E430G Figure 5 shows the transient decrease in T cells (CD3+ cells) after administration of the first dose of IgG1-C-E430G at dose levels ≥4 mg / kg (data cutoff: October 3, 2022). A subsequent increase in peripheral blood T cell count (≥100% increase from baseline) was observed in 7 of 16 patients across 5 of the 6 dose levels, particularly in patients who had not previously received CD38 mAb treatment (Figure 6). Table 17 summarizes the baseline, maximum change, and percentage of peripheral blood T cell counts in patients treated with IgG1-C-E430G.

[0458] Figures 11 and 12 show transient decreases in CD3+CD4+ and CD3+CD8+ T cells after administration of the first dose of IgG1-C-E430G at dose levels ≥4 mg / kg (data cutoff: August 14, 2023). A subsequent increase in peripheral blood CD3+CD4+ T cell counts (≥2 visits, >50% increase from baseline) was observed in 6 of 21 evaluable patients across 5 of the 6 dose levels, particularly in patients who had not previously received treatment with CD38 mAbs (Figure 11). A subsequent increase in peripheral blood CD3+CD8+ T cell counts (≥2 visits, >50% increase from baseline) was observed in 8 of 21 evaluable patients across 5 of the 6 dose levels, particularly in patients who had not previously received treatment with CD38 mAbs (Figure 12). Tables 17 and 18 summarize the baseline, maximum change, and percentage of peripheral blood CD3+ / CD4+ and CD3+ / CD8+ T cell counts in patients treated with IgG1-C-E430G.

[0459] In conclusion, an increase in peripheral T cell counts was observed in a subset of patients administered IgG1-C-E430G across the entire dose level. The observed T cell expansion and proliferation confirm the biological activity of IgG1-C-E430G in vivo and demonstrate its immunomodulatory activity in patients.

[0460] (Table 17) T cells in peripheral blood of patients administered IgG1-C-E430G TIFF2026513656000033.tif193164 Cutoff date: October 3, 2022

[0461] (Table 18) CD4+ T cells in peripheral blood of patients administered IgG1-C-E430G (dose escalation part, data cutoff: August 14, 2023) TIFF2026513656000034.tif173167 Cutoff date: August 14, 2023

[0462] (Table 19) CD8+ T cells in peripheral blood of patients administered IgG1-C-E430G (dose escalation part, data cutoff: August 14, 2023) TIFF2026513656000035.tif167167 Cutoff date: August 14, 2023

[0463] Results: Number of monocytes, B cells, and NKT cells in the peripheral blood of patients treated with IgG1-C-E430G. Monocyte, B cell, and NKT-like cell levels fluctuated in the peripheral blood of patients after IgG1-C-E430G administration; however, consistent or dose-dependent decreases or increases in these lymphocyte populations were not observed in patients treated with IgG1-C-E430G (data not shown).

[0464] In conclusion, the enhancement of CDC activity by IgG1-C-E430G is not associated with a reduction in monocytes, B cells, or NKT-like cells in response to IgG1-C-E430G treatment in a clinical setting.

[0465] Example 6: Evaluation of complement component C2 levels in plasma of patients administered IgG1-C-E430G. method Blood sampling for the analysis of complement C2 in plasma was performed according to the scheme shown in Table 20. Plasma was collected from lavender-top (EDTA) tubes. The tubes were thoroughly mixed and centrifuged at room temperature within 1 hour and 30 minutes of collection. Cell-free plasma was transferred to clean tubes and immediately frozen on dry ice or at -70°C until further use. C2 levels were measured using a radioimmunodiffusion (RID) assay at Quest Diagnostics Nichols Institute (Secaucus, NJ).

[0466] In short, the test samples were gently mixed immediately before use and applied to the wells of a RID plate containing a monospecific antibody against C2 in an agarose gel. After sample application, the plate was tightly closed with the lid and stored flat at room temperature (approximately 20-24°C) for 18-120 hours. To minimize evaporation, the plates were sealed in foil or stored in a sealed plastic box with moistened tissue paper during incubation. The final ring diameter was measured to 0.1 mm at the closest point using a gemstone eyepiece or a digital RID plate reader. The C2 concentration in each test sample was determined by reading directly from the RID reference table and comparing it with the reference curve.

[0467] (Table 20) Schedule of blood sampling for complement analysis TIFF2026513656000036.tif121167 * Dose level 1 (0.2 / 0.6 mg / kg) ** All cohorts except dose level 1 (0.2 / 0.6 mg / kg)

[0468] Results: Plasma complement component C2 levels in patients administered IgG1-C-E430G. Table 21 shows the transient reduction of complement component C2 at all doses, with a median peak reduction from baseline (C1D1, pre-administration) of 64% (range 6%–78%, n=18; patients with C2 levels below LLOQ at baseline were excluded from this analysis), suggesting CDC activity of IgG1-C-E430G in a clinical setting. In most subjects, C2 levels returned to baseline before the next dose, indicating that the treatment did not deplete complement. After the next dose, C2 levels again transiently decreased and returned to baseline before subsequent administrations.

[0469] In conclusion, a decrease in C2 was observed at all dose levels and was transient in most subjects. This confirms the biological activity of IgG1-C-E430G in vivo and demonstrates the CDC activity of IgG1-C-E430G in patients.

[0470] (Table 21) Complement component C2 levels in plasma of patients administered IgG1-C-E430G TIFF2026513656000037.tif175166 Cutoff date: October 3, 2022

[0471] Example 7: Evaluation of complement-mediated lysis activity in the serum of patients administered IgG1-C-E430G. method Blood sampling for the analysis of complement-mediated solubility (CH50) in serum was performed according to the scheme shown in Table 20.

[0472] The complement lysis activity (CH50) in the serum of patients administered IgG1-C-E430G was evaluated by spectrophotometric assay using Autokit CH50 (FUJIFILM Wako, Richmond, VA; catalog number 995-40801). To this end, 10 μL of serum obtained from patients administered IgG1-C-E430G was mixed with 250 μL of liposomes (reagent 1) and incubated at 37°C for 5 minutes. Then, 125 μL of substrate (reagent 2) was added and incubated for another 5 minutes at 37°C. Finally, the absorbance at 340 nm was measured using a Beckman Coulter AU680 system (Brea, CA). An increase in absorbance is proportional to the complement activity in the serum sample.

[0473] Results: Complement-mediated lysis activity in the serum of patients administered IgG1-C-E430G. Table 22 shows the transient decrease in total complement lysis activity (CH50) after the initial dose of IgG1-C-E430G at all evaluable doses (8–24 mg / kg), which was transient in most subjects. The median peak reduction from baseline (C1D1, pre-administration) was 53% (range 2%–92%, n=11). This suggests CDC activity of IgG1-C-E430G in a clinical setting. Complement parameters rapidly returned to baseline levels in most subjects and remained at baseline levels after subsequent administrations, indicating that the therapeutic dose did not deplete complement.

[0474] In conclusion, a transient decrease in complement-mediated lysis activity (CH50) was observed in the peripheral blood of patients administered IgG1-C-E430G at all dose levels evaluated. This confirms the in vivo biological activity of IgG1-C-E430G and demonstrates its CDC activity in the subjects.

[0475] (Table 22) Complement-mediated lysis activity in serum of patients administered IgG1-C-E430G. TIFF2026513656000038.tif167167 Cutoff date: October 3, 2022

[0476] Example 8: Evaluation of cytokine levels in plasma of patients administered IgG1-C-E430G. method Cytokine levels in the plasma of patients administered IgG1-C-E430G were evaluated using a custom-made sandwich immunoassay (V-PLEX Custom Human Biomarkers; Meso Scale Diagnostics, Rockville, AR; catalog number K151A9H-2) with 10-spot MULTI-SPOT® plates pre-coated with capture antibodies against IL-2, IL-6, IL-8, IL-10, TNF-α, and IFN-γ. First, the plates were washed three times with 150 μL / well Wash Buffer (Meso Scale Diagnostics; catalog number R61AA-1). Next, 50 μL / well of plasma sample was added per well and incubated at room temperature for 2 hours with shaking. After incubation, the wells were washed three times with Wash Buffer, and 25 μL / well of SULFO-tagged detection antibodies (anti-IL-2, catalog no. D21QQ; anti-IL-6, catalog no. D21AK; anti-IL-8, catalog no. D21AN; anti-IL-10, catalog no. D21QU; anti-TNF-α, catalog no. D21BH; anti-IFN-γ, catalog no. D21QO; all from Meso Scale Diagnostics) were added. The mixture was incubated at room temperature for 2 hours with shaking. Finally, the wells were washed three times with Wash Buffer, and 150 μL / well of 2×Read Buffer T (Meso Scale Diagnostics; catalog no. R92TC-3) was added. The plates were read using a MESO® QuickPlex SQ 120 SN instrument (Meso Scale Diagnostics; serial no. 1300170726922). The device measures the intensity of synchrotron radiation and provides quantitative measurement of cytokines in plasma samples.

[0477] Results: Cytokine levels in plasma of patients administered IgG1-C-E430G. Figure 7 shows that cytokine levels of IL-2, IL-6, IL-8, IL-10, IFNγ, and TNFα in the plasma of patients administered IgG1-C-E430G remained generally low at all dose levels, with greater variability among individual subjects at higher dose levels (≥16 mg / kg).

[0478] Example 9 - Evaluation of the pharmacokinetics of IgG1-C-E430G in RRMM patients Methods: Determination of pharmacokinetic parameters in patients administered IgG1-C-E430G. The pharmacokinetics of IgG1-C-E430G were evaluated in patients who received IgG1-C-E430G at doses of 0.2 / 0.6 to 24 mg / kg in the dose-escalation part of the first-in-human clinical trial of IgG1-C-E430G (NCT04824794).

[0479] As of the data cutoff date of June 29, 2022, pharmacokinetic data were available from 22 subjects with RRMM (Responsible Life Cycle) who received IgG1-C-E430G in the dose-escalation part (0.2 / 0.6~24 mg / kg) of the first-in-human clinical trial.

[0480] Blood samples for determining serum concentration were collected from patients according to the scheme shown in Table 23. Blood was collected in 4 ml serum separation tubes. After incubation at room temperature (30 minutes) and centrifugation (1500 g for 10 minutes), the serum was transferred to cryotubes and stored at <-65°C.

[0481] (Table 23) Blood sampling schedule. Schedule for a 28-day treatment cycle. Time points relative to infusion; sampling window in parentheses. TIFF2026513656000039.tif154128 1 Cycle 2 only. 2 Cycle 1 only. 3 Cycles 1 and 2 only. 4 Cycles 1-6 only. 5 Only for patients taking 0.2 / 0.6 mg / kg.

[0482] Serum IgG1-C-E430G concentrations were determined using electrochemiluminescence sandwich immunoassay (ECLIA). The assay principle is illustrated in Figure 8. The validated analytical assay range is 0.05 μg / ml to 3.20 μg / ml, and the minimum required dilution (MRD) is 40. IgG1-C-E430G can be quantified up to 990 μg / ml in undiluted serum when additional validated dilution factors are applied.

[0483] Two different anti-idiotype antibodies were used to capture and detect IgG1-C-E430G in human serum. The capture anti-idiotype antibodies were coated onto ECLIA multi-array plates by overnight incubation at 4°C (75 μl / well, diluted to 1 μg / ml with phosphate-buffered saline (PBS, Sigma)). Subsequently, the plates were washed three times with PBS containing 0.01% w / v Tween-20 (Sigma), and then incubated at room temperature for 1 hour with 150 μl / well of Scytek Laboratories Super Block buffer (assay buffer). 75 μl samples (standard, quality control, and test samples) containing IgG1-C-E430G diluted to MRD in assay buffer were added to the plates and incubated at room temperature for 1 hour. The bound IgG1-C-E430G was detected by incubation at room temperature for 1 hour with a SULFO-tagged detection anti-idiotype antibody (75 μl / well, diluted to 1 μg / ml in assay buffer). After incubation, the plate was washed three times and Read Buffer T (MSD) was added. The light emitted by the SULFO tag upon electrochemical stimulation of the electrode surface of the multi-array plate was measured using an MDS multi-array plate reader at 620 nm. The amount of emitted light is an indirect readout of the concentration of IgG1-C-E430G in the sample.

[0484] PK parameters were calculated using the Phoenix 64 software package (version 8.2, Certara USA, Inc., Princeton, NJ) using a non-compartment method consistent with the route of administration (intravenous infusion). The following parameters were derived from the serum concentration-time profiles for administration on day 1 of cycle 1, day 8 of cycle 1 (only for patient E who received 0.2–0.6 mg / kg), and day 1 of cycle 2: AUC 0-t -linear up, log-linear down. Calculated using the trapezoidal method, from the start of administration, the quantifiable concentration (d * Area under the serum concentration-time curve up to the final time point before the next dose at a concentration of μg / mL. • CL-clearance (L / d / kg), dose / (AUC if possible) 0-t +C t / λ z ) is calculated as, where, λ z (d -1 ) is the slope of the logarithmic concentration-time curve determined by regression using at least three observations in the disappearance phase, C t This is the predicted concentration, not the concentration observed at the final point in time at quantifiable concentrations.

[0485] The PK profile includes all PK observations from immediately before administration to the start of the next administration, where a split dose over two consecutive days is considered a single dose. All pre-treatment concentrations, and pre-administration concentrations below the lower limit of quantification (BLQ) at the start of the subsequent PK profile, are imputed with a value of 0. All other BLQ observations were omitted. PK parameters were summarized as mean and standard deviation for each dose and PK profile.

[0486] Results: PK observation results in patients administered IgG1-C-E430G Figure 9 shows that the peak concentration at the end of infusion increased with increasing dose, followed by a two-step decrease. The PK profile at 16 mg / kg was more consistent across subjects, and exposure was better maintained with bi-weekly administration compared to lower dose levels. Peak concentrations showed limited accumulation with weekly administration from day 8 onward at all dose levels, suggesting faster total clearance across dose levels than typical for IgG1 antibody therapies. An increase in pre-administration concentration was observed in some patients between weekly administrations, most pronounced in patients with minimum response or better (e.g., patients F, T, J, C), indicating a reduced impact of time-dependent target-mediated drug elimination due to potential target cell depletion.

[0487] AUC calculated for patients who received IgG1-C-E430G after the first and fifth doses, and after the second dose for patients who received 0.2 / 0.6 mg / kg. 0-t The AUC and CL were plotted against dose in Figure 10 and summarized for each dose level in Table 24. These data represent the AUC after the first and fifth doses. 0-t However, it shows that the increase is greater than the dose-proportionality up to 4 mg / kg, and is almost proportional to the dose above 4 mg / kg. Between subjects administered 16 mg / kg and those administered 24 mg / kg, the AUC 0-t Significant overlap was observed. Initial clotting was rapid at doses below 4 mg / kg and nearly constant at doses above 4 mg / kg. These results suggest the effect of target-mediated drug elimination at weekly administration at dose levels below 4 mg / kg and a higher degree of target saturation at doses above 4 mg / kg. Clotting observed with IgG1-C-E430G at dose levels of 4–24 mg / kg was higher than that observed with previous anti-CD38 antibodies at similar dose levels.

[0488] In summary, the PK of IgG1-C-E430G is consistent with the AUC at doses of 0.2–4 mg / kg, which corresponds to targeted drug elimination. 0-tCharacterized by increases exceeding proportionality and nearly proportional increases at higher dose levels, suggesting a higher degree of target saturation at weekly administrations of ≥ 4 mg / kg doses. CL was faster than typical with IgG1 antibodies and faster than with previous anti-CD38 antibodies at similar dose levels. Some patients showed an increase in pre-administration concentrations between weekly administrations, suggesting target depletion over time.

[0489] (Table 24) PK parameters of patients administered IgG1-C-E430G TIFF2026513656000040.tif226160

[0490] Example 10 - Further evaluation of clinical efficacy by dose escalation Method: Dose escalation - efficacy The clinical trial design for IgG-C-E430G is described in Example 2, and a schematic diagram is shown in Figure 2. In the dose-escalation part of the clinical trial, subjects with RRMM were treated with IgG1-C-E430G at six different dose levels in this first-in-human clinical trial. The reporting period is from October 2022 to the data cutoff date of August 14, 2023.

[0491] result During this reporting period, additional follow-up data were collected for subjects who remained in the dose-escalation phase of the IgG-C-E430G clinical trial. Of the 24 subjects in this phase of the clinical trial, 2 subjects (8.3%) were still receiving clinical trial treatment at the data cutoff date. 13 subjects (54.2%) discontinued treatment but remained participating in the clinical trial. 9 subjects (37.5%) discontinued treatment and either dropped out of the trial or died. The median duration of infusion at the prescribed dose of IgG1-C-E430G was 3.8 hours (range: 1.1–4.6).

[0492] Overall, the efficacy results in the dose escalation phase as of August 14, 2023, were unchanged from those in the previous reporting period, i.e., prior to October 2022. For the best overall efficacy at the six different dose levels administered during that reporting period, please refer to Example 3.

[0493] In summary, the efficacy data for the current period confirm that IgG1-C-E430G has clinical activity in patients with RRMM who are anti-CD38 mAb naive, as well as in patients with RRMM who have a history of treatment with anti-CD38 mAbs.

[0494] Example 11 - Further clinical safety evaluation by dose escalation Method: Dose escalation - safety The clinical trial design for IgG-C-E430G is described in Example 2, and a schematic diagram is shown in Figure 2. In the dose-escalation part of the clinical trial, subjects with RRMM were treated with IgG1-C-E430G at six different dose levels in this first-in-human clinical trial. The reporting period is from October 2022 to the data cutoff date of August 14, 2023.

[0495] result Overall, the safety results for the dose-escalation portion of the IgG-C-E430G clinical trial at the data cutoff date remained unchanged from those for the previous reporting period, i.e., prior to October 2022. For an overview of TEAEs, related TEAEs, serious TEAEs, related serious TEAEs, and AESIs at the six dose levels administered during the previous reporting period, please refer to Example 4.

[0496] In summary, safety data collected from October 2022 to August 14, 2023, confirmed that IgG1-C-E430G has a tolerable safety profile with no events of tumor lysis syndrome or cytokine release syndrome.

[0497] Example 12 - Clinical efficacy evaluation in expanded Part A Method: Effectiveness - Expanded Part A The clinical trial design and preliminary efficacy of IgG1-C-E430G, including expanded Part A, are described in Example 2. Figure 2 shows a schematic diagram of the IgG-C-E430G clinical trial design.

[0498] In this part of the clinical trial, subjects with RRMM who were anti-CD38 mAb naive were treated with IgG1-C-E430G at RP2D identified for RRMM in the dose-escalation part of the clinical trial. As of the data cutoff date of August 14, 2023, efficacy data were available from these subjects who received 16 mg / kg of IgG1-C-E430G in expanded part A.

[0499] result Table 25 shows the best overall response in 11 subjects with RRMM who were anti-CD38 mAb naive and treated with 16 mg / kg of anti-CD38 mAb in this clinical trial. Of the 11 subjects, 8 (72.7%) received two cycles of anti-CD38 mAb. The best overall response among the 11 subjects was complete response (9.1%) in 1 subject, best partial response (18.2%) in 2 subjects, partial response (27.3%) in 3 subjects, minimal response (18.2%) in 2 subjects, stable (9.1%) in 1 subject, and unassessable (18.2%) in 2 subjects.

[0500] (Table 25) Best Overall Effect - Expanded Part A - RRMM Anti-CD38 mAb Naive TIFF2026513656000041.tif139161[a] Based on the Clopper and Pearson Act. [b] ORR includes patients with the best CR and PR effects; clinical benefit includes ORR and MR.

[0501] In summary, efficacy data collected from October 2022 to August 14, 2023, in expanded Part A of the IgG-C-E430G clinical trial demonstrated that IgG1-C-E430G has clinical activity in anti-CD38 mAb naive RRMM patients.

[0502] Example 13 - Clinical Safety Evaluation in Expanded Part A Methods: Expanded Part A - Clinical Safety The clinical trial design for IgG1-C-E430G, including expanded Part A, and the safety evaluation are described in Example 2. Figure 2 shows a schematic diagram of the IgG1-C-E430G clinical trial design.

[0503] In this part of the clinical trial, subjects with RRMM who were anti-CD38 mAb naive were treated with IgG1-C-E430G at RP2D identified for RRMM in the dose-escalation part of the clinical trial. As of the data cutoff date of August 14, 2023, safety data were available from these subjects who received 16 mg / kg of IgG1-C-E430G in expanded part A.

[0504] result As of August 14, 2023, 9 out of 11 subjects (81.8%) in the expanded Part A had experienced at least one study-related adverse event (TEAE) (Table 26). The most common TEAEs reported in this part of the clinical trial (≥20% of subjects) were neutropenia (6 subjects; 54.5%), as well as anemia, headache, IRR, thrombocytopenia, and upper respiratory tract infection (3 subjects for each event; 27.3%). Eight subjects (72.7%) experienced TEAEs considered to be related to IgG1-C-E430G (Table 27). The most frequently reported related TEAEs were neutropenia (6 subjects; 54.5%), infusion reactions (3 subjects; 27.3%), anemia (2 subjects; 18.2%), and thrombocytopenia (2 subjects; 18.2%).

[0505] Severe TEAEs were reported in 5 subjects (45.5%) (Table 28). The most common severe TEAEs were cardiac arrest (2 subjects; 18.2%), as well as anemia, brain injury, death, respiratory tract infection, seizures, and upper respiratory tract infection (1 subject per event; 9.1%). Two subjects (18.2%) experienced severe TEAEs considered to be associated with IgG1-C-E430G: anemia, brain injury, cardiac arrest, and death (1 subject per event; 9.1%) (Table 29).

[0506] Three subjects (27.3%) experienced an internal rate of response (IRR) at a dose level of 16 mg / kg (Table 30).

[0507] No cases of cytokine release syndrome were reported.

[0508] Three subjects (27.3%) discontinued clinical trial treatment but remained in the clinical trial, while five subjects (45.5%) discontinued clinical trial treatment and either dropped out or died. Reasons for discontinuation included disease progression (3 subjects; 27.3%), clinical progression (1 subject; 9.1%), and adverse events (4 subjects; 36.4%). The main reasons for discontinuing the clinical trial were death (4 subjects; 36.4%) and loss to follow-up (1 subject; 9.1%).

[0509] (Table 26) Adverse events occurring during the study treatment - Expanded Part A - RRMM anti-CD38 mAb naive TIFF2026513656000042.tif227161TIFF2026513656000043.tif227161TIFF2026513656000044.tif44161 Percentages are based on N, and multiple adverse events with the same base word are counted only once per subject. This is coded by MedDRA v.26.1. Adverse events that occurred during the study treatment were AEs that began after the first IMP and within 30 days of the last IMP, or pre-existing AEs that worsened.

[0510] (Table 27) Adverse events occurring under relevant study treatments - Expanded Part A - RRMM anti-CD38 mAb naive TIFF2026513656000045.tif198159 Percentages are based on N, and multiple adverse events with the same basic word are counted only once per subject. This is coded by MedDRA v.26.1. Adverse events that occurred during the study treatment were AEs that began after the first IMP and within 30 days of the last IMP, or pre-existing AEs that worsened. Relevance: When the principal investigator records the relevance to IMP, or when the relevance is not recorded.

[0511] (Table 28) Serious adverse events occurring during experimental treatment - Expanded Part A - RRMM anti-CD38 mAb naive TIFF2026513656000046.tif73159 Percentages are based on N, and multiple adverse events with the same base word are counted only once per subject. This is coded by MedDRA v.26.1. Adverse events that occurred during the study treatment were AEs that began after the first IMP and within 30 days of the last IMP, or pre-existing AEs that worsened.

[0512] (Table 29) Related serious adverse events occurring during investigational treatment - Expanded Part A - RRMM anti-CD38 mAb naive TIFF2026513656000047.tif51159 Percentages are based on N, and multiple adverse events with the same basic word are counted only once per subject. This is coded by MedDRA v.26.1. Adverse events that occurred during the study treatment were AEs that began after the first IMP and within 30 days of the last IMP, or pre-existing AEs that worsened. Relevance: When the principal investigator records the relevance to IMP, or when the relevance is not recorded.

[0513] (Table 30) Summary of particularly noteworthy adverse events: Infusion reactions (IRRs) - Expanded Part A - RRMM anti-CD38 mAb naive TIFF2026513656000048.tif88159 Infusion reactions are limited to events marked as AESI, and the basic term is infusion reaction.

[0514] In summary, safety data collected from October 2022 to August 14, 2023, in the expanded Part A clinical trial of IgG-C-E430G demonstrated that IgG1-C-E430G has a manageable safety profile in anti-CD38 mAb naive RRMM patients.

[0515] Example 14 - Evaluation of lymphocyte population in whole blood of patients administered IgG1-C-E430G in Expanded Part A method: Lymphocyte populations in whole blood from RRMM patients administered 16 mg / kg IgG1-C-E430G were evaluated as described in Example 5. As of the data cutoff date of August 14, 2023, preliminary pharmacodynamic data were available from 10 subjects with RRMM who received 16 mg / kg IgG1-C-E430G in expanded Part A of the first-in-human clinical trial.

[0516] result: Peripheral blood NK cell count in patients administered 16 mg / kg of IgG1-C-E430G Figure 13 shows that IgG1-C-E430G administration was associated with a rapid decrease in the number of peripheral blood NK cells (CD3- / CD56+ / CD16+ cells). The median maximum reduction in NK cell count compared to baseline (C1D1, pre-administration) was 204% (range -812% to 83%, n=11) in evaluable patients. In the majority of patients, NK cell counts remained low during IgG1-C-E430G treatment. Table 31 summarizes the baseline, maximum change, and percentage of peripheral blood NK cell counts in patients treated with IgG1-C-E430G.

[0517] In conclusion, similar to the observations in the dose escalation phase (Example 5), a significant reduction in NK cells was observed in patients administered 16 mg / kg of IgG1-C-E430G in expanded Part A. The observed decrease in NK cells confirms the biological activity of IgG1-C-E430G in the patients and indicates the ADCC activity of IgG1-C-E430G.

[0518] (Table 31) Peripheral blood NK cell population of patients administered 16 mg / kg of IgG1-C-E430G TIFF2026513656000049.tif134128 Cutoff date: August 14, 2023

[0519] Results: Peripheral blood T cell count in patients administered 16 mg / kg of IgG1-C-E430G Figures 14 and 15 show the transient decrease in CD3+CD4+ and CD3+CD8+ T cells after administration of the initial dose of IgG1-C-E430G at 16 mg / kg (data cutoff: August 14, 2023). A subsequent increase in peripheral blood CD3+CD4+ T cell counts (≥2 visits, >50% increase from baseline) was observed in 1 out of 10 evaluable patients (Figure 14). In addition, an increase in peripheral blood CD3+CD8+ T cell counts (≥2 visits, >50% increase from baseline) was observed in 3 out of 10 evaluable patients (Figure 15). Tables 32 and 33 summarize the baseline, maximum change, and percentage of peripheral blood CD3+CD4+ and CD3+CD8+ T cell counts in patients treated with IgG1-C-E430G.

[0520] In conclusion, after an initial decrease, an increase in peripheral T cell count was observed in a subset of patients administered 16 mg / kg of IgG1-C-E430G. In particular, CD3+CD8+ T cells showed a significant increase in some subjects. The observed T cell expansion and proliferation confirm the biological activity of IgG1-C-E430G in vivo and demonstrate its immunomodulatory activity in patients.

[0521] (Table 32) CD4+ T cells in peripheral blood of patients administered 16 mg / kg of IgG1-C-E430G TIFF2026513656000050.tif123128 Cutoff date: August 14, 2023

[0522] (Table 33) CD8+ T cells in peripheral blood of patients administered 16 mg / kg of IgG1-C-E430G TIFF2026513656000051.tif123128 Cutoff date: August 14, 2023

[0523] Example 15 - Evaluation of complement-mediated lysis activity in the serum of patients administered 16 mg / kg of IgG1-C-E430G in Expanded Part A. method: The total complement lysis activity (CH50) in serum from RRMM patients administered 16 mg / kg IgG1-C-E430G was evaluated as described in Example 7. As of the data cutoff date of August 14, 2023, preliminary CH50 data were available from 10 subjects with RRMM who received 16 mg / kg IgG1-C-E430G in the expanded Part A of the First-in-Human Clinical Trial.

[0524] Results: Complement-mediated lysis activity in the serum of patients administered IgG1-C-E430G. Table 34 shows the transient decrease in total complement lysis activity (CH50) after the initial dose of IgG1-C-E430G. The median peak reduction from baseline (C1D1, pre-administration) was 60% (range -91% to 40%, n=10). This suggests the CDC activity of IgG1-C-E430G in a clinical setting. Complement parameters rapidly returned to baseline levels in most subjects and remained at baseline levels after subsequent administrations, indicating that the treatment does not deplete complement.

[0525] In conclusion, similar to the observations in the dose escalation phase (Example 7), a transient decrease in complement-mediated lysis activity (CH50) was observed in the peripheral blood of patients administered IgG1-C-E430G. This confirms the biological activity of IgG1-C-E430G in vivo and demonstrates the CDC activity of IgG1-C-E430G in patients.

[0526] (Table 34) Complement-mediated lysis activity in serum of patients administered 16 mg / kg of IgG1-C-E430G. TIFF2026513656000052.tif123128 Cutoff date: August 14, 2023

[0527] Example 16 - Evaluation of cytokine levels in plasma of patients administered IgG1-C-E430G in Expanded Part A. method: Plasma cytokine levels from RRMM patients administered 16 mg / kg IgG1-C-E430G were evaluated as described in Example 8. As of the data cutoff date of August 14, 2023, preliminary pharmacodynamic data were available from 11 subjects with RRMM who received 16 mg / kg IgG1-C-E430G in the expanded Part A of the First-in-Human Clinical Trial.

[0528] result Figure 16 and Table 35 show no significant regulation of cytokine levels of IL-2, IL-6, IL-8, IL-10, IFNγ, and TNFα in the plasma of patients administered 16 mg / kg IgG1-C-E430G, which is consistent with the observations in the dose escalation phase (Example 8).

[0529] (Table 35) Cytokine levels in plasma of patients administered 16 mg / kg of IgG1-C-E430G (Expanded Part A, Data Cutoff: August 14, 2023) TIFF2026513656000053.tif133161 Cutoff date: August 14, 2023

[0530] Example 17 - PK Evaluation in Expanded Part A method: The pharmacokinetics of IgG1-C-E430G were evaluated in patients who received 16 mg / kg of IgG1-C-E430G in the expanded Part A of the first-in-human clinical trial of IgG1-C-E430G.

[0531] As of the data cutoff date of August 14, 2023, pharmacokinetic data were available for 11 subjects with RRMM who received IgG1-C-E430G via expanded Part A, and for 23 subjects with RRMM who received IG-C-E430G via dose escalation.

[0532] The blood sampling schedule and assay methodology for determining serum concentration are the same as those used in dose escalation and are described in Example 9.

[0533] The PK profile included all serum concentrations from all subjects for which PK could be evaluated. All pre-treatment concentrations, and pre-administration concentrations below the lower limit of quantification (BLQ) at the start of subsequent PK profiles, were imputed with values ​​of 0.

[0534] result Figure 17 shows a controlled comparison of concentration-time profiles for subjects receiving a 16 mg / kg dose in dose escalation (A) and expanded cohort A (B). As expected, the PK profiles for subjects receiving similar doses are similar between the two cohorts. The peak and pre-administration concentrations observed in the figure are comparable between the cohorts.

Claims

1. A method for treating or preventing hematological malignancies in a person who needs it, The process includes administering an antibody that binds to human CD38 to a subject in a therapeutically effective dose, The antibody, a. An antigen-binding region comprising VH CDR1 having the sequence described in SEQ ID NO: 2, VH CDR2 having the sequence described in SEQ ID NO: 3, VH CDR3 having the sequence described in SEQ ID NO: 4, VL CDR1 having the sequence described in SEQ ID NO: 6, VL CDR2 having the sequence AAS, and VL CDR3 having the sequence described in SEQ ID NO: 7, and b. Fc region containing mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 in the human IgG monohelic acid (amino acid residues are numbered according to the EU index). The method, including the method described above.

2. The method according to claim 1, wherein the antibody is administered in a dose of at least about 4 mg / kg body weight.

3. The method according to claim 1, wherein the antibody is administered in a dose in the range of approximately 4 mg / kg to approximately 24 mg / kg body weight.

4. The method according to claim 1, wherein the antibody is administered in a dose in the range of approximately 4 mg / kg to approximately 16 mg / kg body weight.

5. The method according to claim 1, wherein the antibody is administered in a dose in the range of approximately 4 mg / kg to approximately 8 mg / kg body weight.

6. The method according to any one of claims 1 to 3, wherein the antibody is administered in a dose in the range of approximately 8 mg / kg to approximately 16 mg / kg body weight.

7. The method according to any one of claims 1 to 5, wherein the antibody is administered at a dose of approximately 4 mg / kg body weight.

8. The method according to any one of claims 1 to 6, wherein the antibody is administered at a dose of approximately 8 mg / kg body weight.

9. The method according to any one of claims 1 to 4 and 6, wherein the antibody is administered at a dose of approximately 16 mg / kg body weight.

10. The method according to any one of claims 1 to 3, wherein the antibody is administered at a dose of approximately 24 mg / kg body weight.

11. The method according to any one of claims 1 to 10, wherein the antibody is administered in a dose of approximately 250 to 2000 mg, for example, approximately 280 to 1700 mg.

12. The method according to any one of the claims, wherein the antibody is administered over approximately 4 weeks or approximately 28 days, for example, in a cycle of 4 weeks or 28 days.

13. The method according to any one of the claims, wherein the antibody is administered weekly (Q1W), preferably the weekly administration is carried out for at least 8 times or 2 cycles.

14. The method according to any one of the claims, wherein the antibody is administered once every two weeks (Q2W - every other week), preferably the every two weeks administration is carried out for at least eight times or for four cycles, and optionally the every two weeks administration follows the weekly administration.

15. The method according to any one of the claims, wherein the antibody is administered once every four weeks (Q4W), preferably, the four-weekly administration is performed at least once, and optionally, the four-weekly administration follows an administration every one or two weeks.

16. The method according to any one of the claims, wherein the antibody is administered in a 28-day (4-week) cycle, with weekly administration in cycles 1 and 2 (Q1W), bi-weekly administration in cycles 3 to 6 (Q2W), and monthly administration from cycle 7 onward (Q4W).

17. The method according to any one of the claims, wherein the antibody is administered over a period of at least two cycles, preferably at least four cycles, more preferably at least six cycles, and even more preferably at least seven cycles.

18. The method according to any one of the claims, wherein at least the initial dose of the antibody is administered as divided doses over two subsequent days, preferably divided into substantially equal amounts.

19. The method according to any one of the claims, wherein the antibody is administered by intravenous injection or infusion.

20. The method according to any one of the claims, wherein the antibody is administered by intravenous injection or infusion in a volume of 100 to 500 ml over a period of 1 to 11 hours, for example, 3 to 10 hours.

21. The method according to any one of the claims, wherein the hematological malignancy is a CD38-positive hematological malignancy or a hematological malignancy known to express CD38, and the antibody is administered for a period of time sufficient to treat the CD38-positive hematological malignancy.

22. The method according to any one of the claims, wherein the hematological malignancy is a cancer that is recurrent or refractory to previous anti-cancer treatments.

23. The method according to any one of the claims, wherein the hematological malignancy is a cancer that is refractory to prior therapies including an anti-CD38 antibody.

24. The method according to any one of claims 1 to 22, wherein the hematological malignancy is a cancer that has recurred after a prior treatment including an anti-CD38 antibody.

25. The method according to claim 23 or 24, wherein the CD38 antibody is daratumumab or isatuximab.

26. The method according to any one of claims 1 to 22, wherein the subject has not previously been treated with an antibody against CD38.

27. The method according to any one of claims 1 to 22 and 26, wherein the subject has not previously been treated with daratumumab and / or isatuximab.

28. The method according to any one of the claims, wherein the hematological malignancy is multiple myeloma (MM).

29. The method according to any one of the claims, wherein the hematological malignancy is recurrent or refractory multiple myeloma (RRMM).

30. The aforementioned relapsed or refractory multiple myeloma is characterized by evidence of disease progression in the subject to the most recent prior treatment regimen, based on the IMWG 2016 criteria for measurable lesions, and the criteria for the aforementioned relapsed or refractory multiple myeloma are, a. Presence of a plasmacytoma confirmed by biopsy or a prior record of monoclonal plasma cells in ≥10% of the bone marrow; b. Measurable lesions at baseline as defined below: i. IgG, IgA, IgD, or IgM myeloma: Serum M protein level ≥ 0.5 g / dL (≥ 5 g / L) or urinary M protein level ≥ 200 mg / 24 hours, ii. Light chain myeloma: Serum Ig free light chain (FLC) ≥ 10 mg / dL and abnormal serum IgκλFLC ratio The method according to claim 29.

31. The method according to any one of claims 1 to 27, wherein the hematological malignancy is diffuse large B-cell lymphoma (DLBCL), for example, relapsed or refractory DLBCL.

32. The method according to any one of the claims, wherein the treatment induces one or more therapeutic effects in the subject, and optionally, the one or more therapeutic effects are improved compared to baseline.

33. The method according to claim 32, wherein one or more of the therapeutic effects are selected from the group consisting of overall response rate, duration of response, and time to response.

34. The method according to claim 32 or 33, wherein the therapeutic effect is a strict complete response, complete response, best partial response, partial response, minimal response, or stable state, and can optionally be continued until the disease progresses or the patient benefit ceases.

35. The method according to any one of the claims, wherein the hematological malignancy is preferably (relapsed or refractory) multiple myeloma, the therapeutic effect is an overall response rate of at least 14% in the treated subjects, and optionally the antibody is administered in a dose of at least (about) 4 mg / kg, for example, (about) to 24 mg / kg.

36. The method according to any one of claims 1 to 22, 26 to 34, wherein the hematological malignancy is a cancer that has not been previously treated with a prior treatment comprising an anti-CD38 antibody, for example daratumumab or isatuximab, preferably multiple myeloma, the therapeutic effect is an overall response rate of at least 40% in the treated subjects, and optionally, the antibody is administered in a dose of at least (about) 4 mg / kg, for example (about) 4 to 24 mg / kg.

37. The method according to any one of claims 1 to 25, 28 to 34, wherein the hematological malignancy is a cancer that has relapsed or refractory to prior anticancer therapies, such as prior therapies including an anti-CD38 antibody such as daratumumab or isatuximab, the therapeutic effect is an overall response rate / objective response rate of at least 6% in the treated subjects, and optionally, the antibody is administered in a dose of (about) 4 mg / kg, at least (about) 4 mg / kg, for example, (about) 4 to 24 mg / kg, for example, (about) 16 mg / kg.

38. The method according to any one of claims 1 to 22, 26 to 34, and 36, wherein the hematological malignancy is a cancer that has not been previously treated with a prior treatment comprising an anti-CD38 antibody, preferably daratumumab or isatuximab, preferably multiple myeloma, and the therapeutic effect is a best partial response (VGPR) of at least 25%, e.g., at least 40%, in the treated subjects, e.g., a complete response (CR) of at least 40%, and optionally, the dose is at least about 4 mg / kg body weight or at least about 8 mg / kg body weight or at least about 16 mg / kg body weight or at least about 24 mg / kg body weight.

39. The method according to any one of claims 1 to 25, 28 to 34, and 37, wherein the hematological malignancy is a cancer that is relapsed or refractory to a prior anticancer treatment, for example, a prior treatment comprising an anti-CD38 antibody, preferably daratumumab or isatuximab, preferably multiple myeloma, and the therapeutic effect is a partial response in at least 6% of the treated subjects, and optionally the dose is at least about 16 mg / kg body weight.

40. The method according to any one of claims 32 to 39, wherein the one or more therapeutic effects are achieved with a dose of at least about 4 mg / kg body weight, at least about 8 mg / kg body weight, at least about 16 mg / kg body weight, or at least 24 mg / kg body weight.

41. The method according to any one of the claims, wherein the subject is treated for the management of cytopenia, for example, neutropenia or thrombocytopenia, for example, grade 3 or grade 4 neutropenia or thrombocytopenia.

42. The method according to any one of the claims, wherein the subject is treated with granulocyte colony-stimulating factor (G-CSF).

43. The method according to any one of the claims, wherein the subject is treated for the management of an infusion reaction (IRR), for example, an IRR of grade 2 or higher.

44. The method according to any one of the claims, wherein the subject is treated by pre-infusion drug therapy before the administration of the antibody and / or post-infusion drug therapy after the administration of the antibody, wherein optionally the pre-infusion drug therapy is administered about 1 to 3 hours before the administration of the antibody and / or the post-infusion drug therapy is administered 2 days after the administration of the antibody.

45. The aforementioned pre-infusion drug therapy includes, optionally, corticosteroids (e.g., methylprednisolone, betamethasone, dexamethasone, triamcinolone, prednisone and / or prednisolone), antihistamines (e.g., diphenhydramine), antipyretics (e.g., paracetamol) and / or leukotriene receptor antagonists (e.g., montelukast), a. The corticosteroid is administered in an amount of approximately 60–100 mg of methylprednisolone or an equivalent dose; b. The diphenhydramine is administered in a dose of approximately 25–50 mg; c. The paracetamol is administered in a dose of approximately 650–1000 mg; and / or d. The montelukast is administered in a dose of approximately 10 mg. The method according to claim 44.

46. The method according to claim 44 or 45, wherein the post-infusion drug therapy comprises a corticosteroid, such as methylprednisolone, betamethasone, dexamethasone, triamcinolone, prednisone and / or prednisolone, and optionally the corticosteroid is administered in a dose of 20 mg of methylprednisolone or an equivalent dose.

47. The method according to any one of the claims, wherein the subject exhibits faster clearance of the antibody compared to a reference antibody that does not contain mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 in human IgG monohelic acid (amino acid residues are numbered according to the EU index) (when administered at a similar, equivalent, or comparable dose).

48. The method according to claim 47, wherein the antibody contains a mutation at position E430, preferably E340G, and the reference antibody does not contain a mutation at position E430 (i.e., is wild-type at the position), preferably the reference antibody contains a wild-type CH3 / Fc region.

49. The method according to claim 47 or 48, wherein the aforementioned earlier clearance occurs at a dose of at least 4 mg / kg body weight.

50. The method according to any one of claims 47 to 49, wherein the clearance is defined as a dose divided by the estimated area under the serum or plasma concentration-time curve from the start of administration to an infinite time.

51. The method according to any one of claims 47 to 50, wherein the reference antibody is an IgG1 antibody that does not contain mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 (amino acid residues are numbered according to the EU index) in the human IgG1 heavy chain, and preferably includes a wild-type CH3 / Fc region.

52. The method according to any one of claims 47 to 51, wherein the antibody contains a mutation at position E430, preferably E430G, and the reference antibody does not contain a mutation at position E430 (is wild-type at the position), and preferably the antibody and the reference antibody are IgG1 antibodies except for any specified mutation.

53. The method according to any one of claims 47 to 52, wherein the reference antibody is daratumumab or isatuximab.

54. The aforementioned antibody a. Inducing activation of the complement system in the subject; b. Inducing depletion of peripheral blood NK cells in the subject; and / or c. In the subject described above, induce the expansion and proliferation of peripheral blood T cells, The method according to any one of the above claims.

55. The method according to any one of the claims, wherein the treatment induces activation of the complement system in the subject, and optionally, the activation of the complement system is reflected by a (transient) reduction of complement component C2 in peripheral blood and / or a (transient) reduction of the total complement-mediated lysis activity (CH50).

56. The method according to claim 55, wherein the C2 level is reduced by at least 30% from the baseline, for example, at least (about) 35%, 40%, 45%, 50%, 55%, 58%, 60%, or 64%, and / or the CH50 level is reduced by at least 20% from the baseline, for example, at least about 25%, 30%, 32%, 35%, 40%, 45%, 48%, 50%, 53%, 55%, or 60%.

57. The method according to any one of claims 54 to 56, wherein the treatment induces complement system activation to a range exceeding that of a reference antibody that does not contain mutations in one or more amino acid residues selected from the group corresponding to E430, E345 and S440 in the human IgG1 heavy chain (the amino acid residues are numbered according to the EU index), for example, a reference antibody containing the wild-type Fc region (when administered at a similar or equivalent dose), and optionally, the reference antibody is daratumumab.

58. The method according to any one of claims 54 to 57, wherein the activation / consumption of the complement system and / or the decrease in C2 and / or CH50 are transient and optionally return to baseline within approximately 8 days.

59. The method according to any one of the claims, wherein the NK cell depletion is induced when the antibody is administered at a dose level of at least 0.2 mg / kg and is maintained during treatment.

60. The method according to any one of claims 54 to 49, wherein T cell expansion and proliferation are induced in a subject who has not previously received a CD38 antibody, such as daratumumab or isatuximab, preferably daratumumab.

61. The method according to any one of the claims, wherein the treatment does not result in a substantial dose-dependent increase in plasma levels of pro-inflammatory cytokines, such as IL-2, IL-6, IL-8, IL-10, IFNγ, and / or TNFα, in the subject.

62. The method according to any one of the claims, wherein the treatment does not induce a dose-dependent reduction of B cells, T cells, monocytes and / or NKT-like cells in the subject.

63. The aforementioned antibody a. The subject has an inhibitory effect on CD38 cyclase activity; b. In the subjects described above, induce complement-dependent cell-mediated cytotoxicity (CDC) in cells expressing human CD38; c. In the subjects described above, antibody-dependent cell-mediated injury (ADCC) is induced in cells expressing human CD38; d. In the subjects described above, induce antibody-dependent cell phagocytosis (ADCP) in cells expressing human CD38; e. In the subject, induce apoptosis in the presence of FcgR-containing cells; f. Induce trogocytosis in cells expressing human CD38; or Any combination of ga to f. The method according to any one of the above claims.

64. The method according to any one of the claims, wherein the antibody induces a reduction in the trogocytosis-mediated activity of CD38 on CD38-expressing tumor cells in the subject.

65. The method according to any one of the claims, wherein the antibody induces a reduction in the trogocytosis-mediated activity of CD38 on CD38-expressing immune cells in the subject.

66. The method according to claim 65, wherein the CD38-expressing immune cells are CD38-expressing immunosuppressive cells, and preferably, the reduction of trogocytosis-mediated activity of CD38 on the CD38-expressing immunosuppressive cells reduces their immunosuppressive activity.

67. The method according to claim 66, wherein the CD38-expressing immunosuppressive cells include regulatory T cells (Treg), regulatory B cells (Breg), myeloid-derived suppressor cells (MDSC), immunosuppressive NK cells, immunosuppressive NKT cells, immunosuppressive antigen-expressing cells (APC), immunosuppressive macrophages, or any combination of two or more thereof, preferably Treg.

68. The method according to any one of claims 54 to 67, wherein one or all of a., b., and f. are higher (when administered at similar, equivalent, or comparable doses) compared to a reference antibody that does not contain mutations in one or more amino acid residues selected from the group corresponding to E430, E345, and S440 (amino acid residues are numbered according to the EU index) in the human IgG monohelic acid.

69. The method according to claim 68, wherein the antibody contains a mutation at position E430, preferably E430G, and the reference antibody does not contain a mutation at position E430 (is wild-type at the position), and preferably the antibody and the reference antibody are IgG1 antibodies, except for any specified mutation.

70. The method according to claim 68 or 69, wherein the reference antibody is daratumumab or isatuximab.

71. The aforementioned antibody Variable heavy chain (VH) region containing an amino acid sequence that includes SEQ ID NO: 1 or has at least 80%, e.g., 90%, 95%, 97%, 98%, or 99% identity to SEQ ID NO:

1. The method according to any one of the claims, including

72. The aforementioned antibody Variable light chain (VL) region containing an amino acid sequence that includes SEQ ID NO: 5 or has at least 80%, e.g., 90%, 95%, 97%, 98%, or 99% identity to SEQ ID NO:

5. The method according to any one of the claims, including

73. The aforementioned antibody A variable heavy chain (VH) region different from SEQ ID NO: 1 due to mutations of 12 or fewer amino acid residues, e.g., 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue, e.g., substitution, insertion, or deletion. The method according to any one of the claims, including

74. The aforementioned antibody Variable light chain (VL) regions different from SEQ ID NO: 5 due to mutations of 12 or fewer amino acid residues, e.g., 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue, e.g., substitution, insertion, or deletion. The method according to any one of the claims, including

75. The method according to any one of the claims, wherein the antibody comprises a variable heavy chain (VH) region containing the sequence SEQ ID NO: 1 and a variable light chain (VL) region containing the sequence SEQ ID NO:

5.

76. The method according to any one of the claims, wherein the mutation in the one or more amino acid residues is selected from the group consisting of E430G, E345K, E430S, E430F, E430T, E345Q, E345R, E345Y, S440Y, and S440W, preferably E430G, E345K, E430S, and E345Q.

77. The method according to any one of the claims, wherein the mutation in the one or more amino acid residues comprises E430G.

78. The method according to any one of the claims, wherein the mutation in one or more amino acid residues consists of E430G.

79. The Fc region is One or more further mutations that do not reduce complement-dependent cell-mediated injury (CDC) and / or antibody-dependent cell-mediated injury (ADCC) induced by the antibody variant having one or more further mutations The method according to any one of the claims, including

80. The method according to claim 79, wherein the one or more further mutations are 12 or fewer, for example, mutations of 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid residue, for example, substitution, insertion, or deletion.

81. The method according to any one of the claims, wherein the Fc region is a human IgG1, IgG2, IgG3, or IgG4 isotype, or a mixture thereof, excluding the described mutation.

82. The method according to any one of the claims, wherein the variant Fc region is a human IgG1 Fc region, excluding the described mutation.

83. The method according to any one of the claims, wherein the Fc region is a human IgG1m(f), IgG1m(a), IgG1m(x), IgG1m(z) allotype, or any two or more mixed allotypes thereof, excluding the described mutations.

84. The method according to any one of the claims, wherein the antibody is a human antibody, except for the described mutation.

85. The method according to any one of the claims, wherein the antibody is an IgG1 antibody, except for the described mutation.

86. The method according to any one of the claims, wherein the antibody is a human monoclonal full-length bivalent IgG1m(f), κ antibody, except for the described mutation.

87. The method according to any one of the claims, wherein the CH region is a human IgG1m(f), IgG1m(a), IgG1m(x), and IgG1m(z) allotype, or any two or more mixed allotypes thereof.

88. The method according to any one of the claims, wherein the CH region comprises the sequence of SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, or SEQ ID NO: 45, excluding the described mutation.

89. The method according to any one of the claims, wherein the CH region comprises one or more further mutations.

90. The method according to any one of the claims, wherein Lys(K) at position 447 according to EU numbering is missing.

91. The method according to any one of the claims, wherein the CH region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 24 to SEQ ID NO: 33 and SEQ ID NO:

46.

92. The method according to any one of the claims, wherein the CH region includes SEQ ID NO: 24 or SEQ ID NO: 46, and optionally the light chain includes CL including SEQ ID NO:

37.

93. The method according to any one of the claims, wherein the antibody is a bivalent antibody.

94. The method according to any one of the claims, wherein the antibody is a full-length antibody.

95. The method according to any one of the claims, wherein the antibody is a monoclonal antibody.

96. The method according to any one of the claims, wherein the antibody is a monospecific antibody.

97. The method according to any one of claims 1 to 96, wherein the antibody is a bispecific antibody.

98. The method according to any one of the claims, wherein the antibody is contained in a composition further comprising a pharmaceutically acceptable carrier.

99. The aforementioned antibody a) The antibody at a concentration of 1 to 200 mg / mL, optionally; b) 5-40 mM histidine or acetate; c) 100–400 mM sorbitol or sucrose; and d) Surfactants The method according to any one of the claims, comprising a composition containing the above.

100. The aforementioned antibody It has a pH of approximately 6, and Optionally, in aqueous solution state, a) The antibody at approximately 20 mg / mL, b) Approximately 20 mM histidine, c) Approximately 250 mM sorbitol, and d) Polysorbate 80 at approximately 0.04% w / v To include, consist of, or essentially be made from The method according to any one of the claims, comprising a composition containing the above.

101. The method according to any one of the claims, wherein the antibody is in the state of a composition that is a concentrate to be diluted, for example, with 0.9% NaCl (physiological saline) or dextrose solution, optionally with 5% w / v dextrose solution.

102. An antibody according to any one of the claims, for use in the prevention or treatment of hematological malignancies according to any one of the claims 1 to 101.

103. Use of the antibody according to any one of the claims for the manufacture of a medicament for the prevention or treatment of hematological malignancies according to any one of the claims 1 to 101.

Images