RU2810953C2 - Subcutaneous dosing of anti-cd38 antibodies - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: inventions relate to immunotherapy for CD38-associated cancer. A method and unit dosage form are provided for treating relapsed/refractory multiple myeloma (RRMM) in a subject. The subject is administered subcutaneously isolated human anti-CD38 antibody at a dose of 45 to 1,800 mg, where the dose is a once-weekly dose. The anti-CD38 antibody contains CDR1-6 having the following amino acid sequences SEQ ID NO: 3-8, respectively. The unit dosage form contains from 45 to 1,800 mg of isolated antibody having a heavy chain variable region containing the following SEQ ID NO: 9, and a light chain variable region containing the following SEQ ID NO: 10. The isolated antibody binds to CD38 and does not bind to human red blood cells. The volume of the unit dosage form is 2.5 ml or less.

EFFECT: invention provides effective treatment for RRMM, including in patients with late stage of RRMM preventing side effects.

16 cl, 26 dwg, 11 tbl, 3 ex

Description

Cross reference to related applications

[0001] This application claims priority to US Provisional Application No. 62/649,489, filed March 28, 2018, which is incorporated herein by reference in its entirety.

Technical field

[0002] Disclosed herein are methods and compositions for administering isolated anti-CD38 antibodies via subcutaneous (SC) administration.

State of the art

[0003] CD38, also known as cyclic ADP-ribose hydrolase, is a type II transmembrane glycoprotein with a long C-terminal extracellular domain and a short N-terminal cytoplasmic domain. CD38 is a member of a group of related membrane-bound or soluble enzymes that includes CD157 and the Aplysia ADPR cyclase. This family of enzymes has the unique ability to convert NAD into cyclic ADP-ribose or nicotinic acid adenine dinucleotide phosphate. CD38 is involved in Ca ²⁺ mobilization and signal transduction through tyrosine phosphorylation of numerous signaling molecules, including phospholipase Cγ, ZAP-70, syk, and c-cbl. Based on these observations, CD38 is an important signaling molecule in the maturation and activation of lymphoid cells during their normal development. Among hematopoietic cells, a set of functional effects have been attributed to CD38-mediated signaling, including lymphocyte proliferation, cytokine release, regulation of B and myeloid cell development and survival, and induction of dendritic cell (DC) maturation.

[0004] CD38 is expressed in immature hematopoietic cells, decreased in mature hematopoietic cells, and re-expressed at high levels in activated lymphocytes and plasma cells. For example, CD38 expression at high levels is observed in activated B cells, plasma cells, activated CD4+ T cells, activated CD8+ T cells, NK cells, NKT cells, mature DCs and activated monocytes (see, for example, US Pat. No. 8362211).

[0005] The presence of CD38 autoantibodies has been associated with a number of diseases, including diabetes, chronic autoimmune thyroiditis, and Graves' disease (see Antonelli et al. (2001) Clin. Exp. Immunol. 126: 426-431; Mallone et al. (2001) ) Diabetes 50: 752 and Antonelli et al (2004) J Endocrinol Invest 27: 695-707).

[0006] Increased expression of CD38 has been documented in various diseases, including autoimmune diseases and cancer. Such diseases include systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD) and ulcerative colitis (UC). In patients with RA, plasma cells are increased in joint tissue compared to controls. In patients with SLE, plasmablasts increase in the peripheral blood of patients with more active disease. Current CD20-based B cell therapies, such as rituximab, effectively deplete the CD20+ B cell population but cannot directly and effectively kill plasma cells or plasmablasts because they do not express CD20. Consistent with this idea, patients with RA or SLE with high levels of plasma cells or plasmablasts are unlikely to receive significant clinical benefit from CD20-based therapy. Thus, therapeutics targeting CD38, which is highly expressed on plasma cells and plasmablasts, as well as NK cells and activated T cells, may provide effective treatment for RA and SLE, as well as other diseases characterized by CD-38 expression.

[0007] In particular, increased expression of CD38 has been documented in various diseases of hematopoietic origin, as well as in cell lines derived from them, and has been described as a negative prognostic marker in hematological malignancies. Such diseases include, but are not limited to, multiple myeloma (MM), chronic lymphoblastic leukemia, B-cell chronic lymphocytic leukemia (B-CLL), including B-cell acute lymphocytic leukemia, B- and T-acute lymphocytic leukemia (ALL), acute lymphoblastic leukemia, Waldenström's macroglobulinemia, mantle cell lymphoma, prolymphocytic/myelocytic leukemia, acute myeloid leukemia (AML), chronic myeloid leukemia (CML), follicular lymphoma, NK-cell lymphocytic leukemia, plasmatic leukemia, non-Hodgkin's lymphoma, (NHL), Burkitt's lymphoma (HL), T-cell lymphoma (TCL), hairy cell leukemia (HCL) and Hodgkin lymphoma (HL). In addition, CD38 expression is a prognostic indicator for patients with conditions such as B-CLL et al . (2002) Leukemia 16: 30-35 and Morabito et al . (2001) Leukemia Res. 25: 927-932) and acute myeloid leukemia (Keyhani et al . (1999) Leukemia Res. 24: 153-159). Thus, CD38 represents a useful target in the treatment of diseases of the hematopoietic system.

[0008] Several anti-CD38 antibodies are in clinical trials for the treatment of CD38-associated cancers. However, all prior art anti-CD38 therapeutic antibodies bind to red blood cells (RBCs) and platelets, resulting in higher dosage requirements due to reduced non-productive binding to RBCs. Although CD38 is expressed on red blood cells at levels that are approximately 1000 times lower than on myeloma cells (deWeers et al. (2011) J. Immunol. 186(3):1840-1848), in the blood of patients with MM (multiple myeloma) with active disease, there are approximately 36,000 red blood cells per myeloma cell (Witzig et al. (1993) Cancer 72(1): 108-113). Thus, erythrocytes express 36 times more CD38 molecules than tumor cells. Thus, current anti-CD38 antibody treatments require intravenous administration due to the high doses required for effectiveness beyond red blood cell binding. For example, daratumumab (anti-CD38 IgG1 mAb; DARZALEX® FDA approved and commercially available from Janssen Oncology) requires a very high dose (≥16 mg/kg) and an intensive regimen (8 times a week, 8 times every 2 weeks, then once a month ) for optimal antitumor activity (Xu et al. (2017) Clin. Pharmacol. Ther. 101(6): 721-724).

[0009] Accordingly, treatment using anti-CD38 antibodies that bind to red blood cells is currently focused on intravenous administration due to the large volume of antibody required to achieve therapeutic efficacy, as such large volumes are not suitable for subcutaneous administration. For example, daratumumab should not be administered in low volumes because ≥16 mg/kg is required to achieve target saturation (eg, approximately 1120 mg in a 70-kg patient). The highest known concentration of the drug for subcutaneous administration is 200 mg/ml (Cimzia®, also called certolizumab pegol). Using the highest known subcutaneous drug concentration of 200 mg/mL, daratumumab would have a minimum predicted injection volume of 5.6-11.2 mL, which is a very large volume for subcutaneous administration. Given this large volume and concentration limits, daratumumab must be administered subcutaneously in a volume of 15 mL along with hyaluronidase to promote dispersion and absorption.

[0010] Another anti-CD-38 antibody, isatuximab (commercially available from Sanofi Genzyme and currently in phase 3 clinical trials), is administered at 10 mg/kg and 20 mg/kg in phase 3 trials, corresponding to 700-1400 mg per 70 kg patient. Again, using the highest known subcutaneous drug concentration of 200 mg/mL, the predicted injection volume of isatuximab is 3.5–14 mL.

[0011] In addition to the higher doses and volumes required of currently clinically known anti-CD38 antibodies, their targeting of red blood cells and platelets can cause serious side effects, such as, for example, hemolytic anemia, a condition in which red blood cells are destroyed faster than they are replaced. In an open-label, peer-to-peer study, isatuximab was administered intravenously to 97 patients at a dose of 3 mg/kg every 2 weeks (Q2W; n=23), 10 mg/kg Q2W for 2 cycles followed by Q4W (n=25), 10 mg/kg Q2W ( n=24) and 20 mg/kg every week for 4 doses (1 cycle) followed by Q2W (n=25). The most common severe adverse event (grade 3/4) was anemia, affecting 24% of patients (see http://www.onclive.com/conference-coverage/asco-2016/isatuximab-monotherapy-effective-for-heavily- pretreated-myeloma, the 2016 ASCO Annual Meeting as well as Richter et al (2016) J Clin Oncol 34 (suppl): abstr 8005). In addition to severe anemia, thrombocytopenia and neutropenia are also common severe adverse events (22.9%, 18.4% and 18.4%, respectively; Dimopoulos et al. (2018) Blood 132 (suppl. 1): ASH abstract 155/ oral presentation). Likewise, low blood cell counts (white blood cells, red blood cells, and platelets), anemia, and thrombocytopenia are well-known serious adverse reactions to daratumumab. In one study of daratumumab, 45% of all patients had anemia (19% of which were grade 3) and 48% of patients experienced thrombocytopenia (10% of which were grade 3 and 8% of which were grade 4) (see eg, Darzalex (daratumumab) prescribing information. Horsham, Pennsylvania: Janssen Biotech, Inc. 2018; and review article by Costello (2017) Ther. Adv. Hematol. 8(1): 28-37). In addition, intravenous administration of therapeutic monoclonal antibodies may result in severe infusion-related reactions (SIRs). Common RSIs include but are not limited to nasal congestion, cough, allergic rhinitis, throat irritation, shortness of breath, chills, nausea, hypoxia, hypertension, etc. (Usmani et al. (2016) Blood 128(1): 37-44). With daratumumab, 48% of patients experience RSI after the first dose of treatment ( Usmani et al. (2016) Blood 128(1): 37-44) with 3% severe cases (Darzalex (daratumumab) prescribing information. Horsham, Pennsylvania: Janssen Biotech, Inc 2018). Similarly, RSIs were reported in 40.4% of patients receiving isatuximab, with 4.6% of patients experiencing severe disease ( Dimopoulos et al. (2018) Blood 132: (suppl 1) ASH abstract 155/ oral presentation ) . Therefore, patients receiving isatuximab or daratumumab should be closely monitored for life-threatening and other serious side effects .

[0012] AB79 is a fully human IgG1 immunoglobulin monoclonal antibody that specifically binds to CD38 with high affinity (Kd=6.1×10 ^-10 M). AB79 inhibits the growth of tumor cells expressing CD38 by depleting the cell population through antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). AB79 also reduces the levels of plasma cells and plasmablasts in blood isolated from healthy subjects and patients with systemic lupus erythematosus (SLE) (PCT Application No. PCT/US2017/042128). In healthy cynomolgus monkeys, depletion efficiency for each cell type was positively correlated with CD38 expression level and AB79 dose level (PCT Application No. PCT/US2017/042128). Additionally, AB79 has demonstrated anti-inflammatory and disease-modifying activity in a monkey model of rheumatoid arthritis (US Patent No. US 8362211).

[0013] Given that many anti-CD38 antibodies in the clinic bind red blood cells and are therefore unsuitable for subcutaneous administration and have dangerous side effects, there remains a need in the art for subcutaneous antibody preparations that are safer, more convenient, and more effective for treatment of diseases in the specified binding to CD38, for example in autoimmune diseases and hematological forms of malignant tumors.

The essence of the invention

[0014] Provided herein are methods for treating diseases in which binding to CD38 is indicated, such as, for example, autoimmune diseases and hematologic malignancies, involving subcutaneous administration of isolated anti-CD38 antibodies.

[0015] In one aspect, the present invention provides a method of treating a disease in a subject that is shown to bind to CD38, the method comprising the step of administering subcutaneously to a subject suffering from a disease that is shown to bind to CD38 a therapeutically effective amount of an isolated human anti-CD38 antibody sufficient to treat the disease, wherein the anti-CD38 antibody comprises a heavy chain variable region (VH) comprising a CDR1 having the amino acid sequence of SEQ ID NO: 3, a CDR2 having the amino acid sequence of SEQ ID NO: 4, and a CDR3 having the amino acid sequence of SEQ ID NO: 3 ID NO: 5, or variants of these sequences with up to three amino acid substitutions; and a light chain variable region (VL) comprising CDR1 having the amino acid sequence of SEQ ID NO: 6, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 8 or variants of these sequences containing up to three amino acid substitutions, and the anti-CD38 antibody is administered at a dose of 45 to 1800 milligrams.

[0016] In another aspect, the present invention provides a method of treating a disease in a subject that is shown to bind to CD38, the method comprising the step of administering subcutaneously to a subject suffering from a disease that is shown to bind to CD38, a therapeutically effective amount of an isolated human antibody to CD38 sufficient to treat the disease, wherein the anti-CD38 antibody comprises a VH chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 3, a CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 5 , or variants of these sequences containing up to three amino acid substitutions; and a VL chain region comprising CDR1 having the amino acid sequence of SEQ ID NO: 6, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 8, or sequence variants containing up to three amino acid substitutions, in this case, the antibody to CD38 is administered in a dose of 45 to 1800 milligrams.

[0017] In another aspect, the present invention provides a method of treating a disease in a subject in which binding to CD38 is shown, the method comprising the step of administering subcutaneously to a subject suffering from a disease in which binding to CD38 is shown a therapeutically effective amount of an isolated human antibody to CD38 sufficient to treat the disease, wherein the anti-CD38 antibody comprises a VH chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 3, a CDR2 having the amino acid sequence of SEQ ID NO: 4 and a CDR3 having the amino acid sequence of SEQ ID NO: 5 ; and a VL chain region containing CDR1 having the amino acid sequence of SEQ ID NO: 6, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 8, wherein the anti-CD38 antibody is administered at a dosage of 45 up to 1800 milligrams.

[0018] In one aspect, an anti-CD38 antibody described herein does not cause hemolytic anemia or thrombocytopenia.

[0019] In one aspect, administration of anti-CD38 antibody therapy results in less than 60%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of grade 3 or 4 cases of one or more treatment-emergent adverse events (TEAEs) or treatment-emergent adverse events treatment-related events (TEAEs), selected from the group consisting of anemia, hemolytic anemia, neutropenia, thrombocytopenia, fatigue, infusion-related reactions (IRR), leukopenia and lymphopenia. A TEAE is an adverse event that is observed or diagnosed approximately 30 days after the last dose of the drug, regardless of the cause. TEAE may have any underlying disease- or treatment-related cause that is not related to or related to anti-CD38 antibody, and may be specifically related to anti-CD38 antibody. Accordingly, administration of an anti-CD38 antibody may result in less than 30% incidence of one or more grade 3 or 4 treatment-emergent adverse events (TEEs) selected from the group consisting of anemia, hemolytic anemia, thrombocytopenia, fatigue, reactions infusion-related (IRA), leukopenia and lymphopenia.

[0020] In one aspect, administration of anti-CD38 antibody therapy results in less than 60%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% of grade 3 or 4 cases of one or more treatment-emergent adverse events (TEAEs), selected from the group consisting of from anemia, hemolytic anemia, neutropenia, thrombocytopenia, fatigue, infusion-related reactions (IRR), leukopenia and lymphopenia. A TEA is an adverse event in which the treating physician believes that there is a possible cause-and-effect relationship between the drug used in treatment and the adverse event. Thus, NUCL is thought to be specifically associated with anti-CD38 antibody. Accordingly, administration of an anti-CD38 antibody may result in less than 30% of the incidence of one or more grade 3 or 4 TEA selected from the group consisting of anemia, hemolytic anemia, thrombocytopenia, fatigue, infusion-related reactions (IRR), leukopenia and lymphopenia.

[0021] In one aspect, administration of anti-CD38 antibody therapy causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1 % depletion of red blood cell population.

[0022] In one aspect, administration of anti-CD38 antibody therapy causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1 % platelet depletion.

[0023] In one aspect, the disease is an autoimmune disease or cancer.

[0024] In one aspect, the autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), ulcerative colitis (UC), systemic light chain amyloidosis, and graft-versus-host disease.

[0025] In one aspect, the hematologic malignancy is selected from the group consisting of multiple myeloma, chronic lymphoblastic leukemia, chronic lymphocytic leukemia, plasma cell leukemia, acute myeloid leukemia, chronic myeloid leukemia, B-cell lymphoma, and Burkitt's lymphoma.

[0026] In one aspect, the hematologic malignancy is multiple myeloma.

[0027] In another aspect, the autoimmune disease is systemic light chain amyloidosis.

[0028] In one aspect, the VH chain region contains an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 10 Accordingly, the VH chain region may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 9, and the VL chain region may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 10. Accordingly , the VH chain region may comprise an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 9, and the VL chain region may comprise an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 10. Accordingly, the region the VH chain region may contain an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 10. Accordingly, the VH chain region may comprise an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 10. Accordingly, the VH chain region may comprise an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 10.

[0029] Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 80% sequence identity to SEQ ID NO: 9, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 80% sequence identity to SEQ ID NO: 10 Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 85% sequence identity to SEQ ID NO: 9. and the VL chain may comprise the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 85% sequence identity to SEQ ID NO: 10. Accordingly , the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 90% sequence identity to SEQ ID NO: 9, and the chain VL may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 90% sequence identity to SEQ ID NO: 10. Accordingly, the chain VH may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 95% sequence identity to SEQ ID NO: 9, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 95% sequence identity with SEQ ID NO: 10. Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 97% sequence identity with SEQ ID NO: 9, and the VL chain may contain the sequences CDRs defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 97% sequence identity with SEQ ID NO: 10. Accordingly, the VH chain may contain the sequences CDRs defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 99% sequence identity to SEQ ID NO: 9, and the VL chain may contain CDR sequences, defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 99% sequence identity with SEQ ID NO: 10.

[0030] In one aspect, the VH chain region has the amino acid sequence SEQ ID NO: 9 or a variant thereof with up to three amino acid substitutions, and the VL chain region has the amino acid sequence SEQ ID NO: 10 or a variant thereof with up to three amino acid substitutions.

[0031] In one aspect, the VH chain region has the amino acid sequence of SEQ ID NO: 9, and the VL chain region has the amino acid sequence of SEQ ID NO: 10.

[0032] In one aspect, the VH chain region contains an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 12 Accordingly, the VH chain may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 11, and the VL chain region may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 12.

[0033] Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 80% sequence identity to SEQ ID NO: 11, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 80% sequence identity to SEQ ID NO: 12 Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 85% sequence identity to SEQ ID NO: 11. and the VL chain may comprise the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 85% sequence identity to SEQ ID NO: 12. Accordingly , the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 90% sequence identity to SEQ ID NO: 11, and the chain VL may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 90% sequence identity to SEQ ID NO: 12. Accordingly, the chain VH may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 95% sequence identity to SEQ ID NO: 11, and the VL strand may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 95% sequence identity with SEQ ID NO: 12. Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 97% sequence identity with SEQ ID NO: 11, and the VL chain may contain the sequences CDRs defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 97% sequence identity with SEQ ID NO: 12. Accordingly, the VH chain may contain the sequences CDRs defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 99% sequence identity to SEQ ID NO: 11, and the VL chain may contain CDR sequences, defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 99% sequence identity with SEQ ID NO: 12.

[0034] In one aspect, an anti-CD38 antibody comprises the heavy chain amino acid sequence SEQ ID NO: 11 or a variant thereof having up to three amino acid substitutions and the light chain amino acid sequence SEQ ID NO: 12 or a variant thereof having up to three amino acid substitutions.

[0035] In one aspect, the anti-CD38 antibody comprises the heavy chain amino acid sequence of SEQ ID NO: 11 and the light chain amino acid sequence of SEQ ID NO: 12.

[0036] In one aspect, a therapeutically effective amount is a dose of 45 to 1800 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 45 to 1200 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 45 to 600 milligrams. Accordingly, a therapeutically effective amount may be a dose of 45 to 135 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 135 to 1800 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 135 to 1200 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 135 to 600 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 600 to 1800 milligrams. Accordingly, a therapeutically effective amount may be a dose of 600 to 1200 milligrams. Accordingly, a therapeutically effective amount may be a dose of 1,200 to 1,800 milligrams.

[0037] In one aspect, the human anti-CD38 antibody is administered in the form of a pharmaceutically acceptable composition.

[0038] In another aspect, the present invention provides a method of treating a hematologic malignancy in a subject, the method comprising the step of administering subcutaneously to a subject suffering from a hematologic malignancy a therapeutically effective amount of an isolated human anti-CD38 antibody sufficient to treat the hematologic malignancy, wherein an anti-CD38 antibody contains a VH chain region containing CDR1 having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 5, or variants of these sequences having up to three amino acid substitutions; and a VL chain region comprising CDR1 having the amino acid sequence SEQ ID NO: 6, CDR2 having the amino acid sequence SEQ ID NO: 7, and CDR3 having the amino acid sequence SEQ ID NO: 8 or variants of these sequences containing up to three amino acid substitutions, and wherein the antibody is administered at a dose of from 45 to 1800 milligrams.

[0039] In another aspect, the present invention provides a method of treating a hematologic malignancy in a subject, the method comprising the step of administering subcutaneously to a subject suffering from a hematologic malignancy a therapeutically effective amount of an isolated human anti-CD38 antibody sufficient to treat the hematologic malignancy, at This anti-CD38 antibody contains a VH chain region containing CDR1 having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 4 and CDR3 having the amino acid sequence of SEQ ID NO: 5, or variants of these sequences containing up to three amino acid substitutions; and a VL chain region comprising CDR1 having the amino acid sequence of SEQ ID NO: 6, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 8, or sequence variants containing up to three amino acid substitutions, in this case, the antibody to CD38 is administered in a dose of 45 to 1800 milligrams.

[0040] In another aspect, the present invention provides a method of treating a hematologic malignancy in a subject, the method comprising the step of administering subcutaneously to a subject suffering from the hematologic malignancy a therapeutically effective amount of an isolated human anti-CD38 antibody sufficient to treat the hematologic malignancy, at wherein the anti-CD38 antibody comprises a VH chain region containing CDR1 having the amino acid sequence of SEQ ID NO: 3, CDR2 having the amino acid sequence of SEQ ID NO: 4 and CDR3 having the amino acid sequence of SEQ ID NO: 5; and a VL chain region containing CDR1 having the amino acid sequence of SEQ ID NO: 6, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 8, wherein the anti-CD38 antibody is administered at a dosage of 45 up to 1800 milligrams.

[0041] In one aspect, the anti-CD38 antibody does not cause hemolytic anemia or thrombocytopenia.

[0042] In one aspect, administration of an anti-CD38 antibody results in less than 60%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10 %, less than 5%, less than 4%, less than 3%, less than 3% or less than 1% of grade 3 or 4 cases of one or more treatment-emergent adverse events (TEAEs) or TEAEs selected from the group, consisting of anemia, including hemolytic anemia, neutropenia, thrombocytopenia, fatigue, infusion-related reactions (IRR), leukopenia and lymphopenia. Accordingly, administration of an anti-CD38 antibody may result in less than 30% incidence of one or more treatment-emergent adverse events or grade 3 or 4 TEAEs selected from the group consisting of anemia, hemolytic anemia, thrombocytopenia, fatigue, reactions, associated with infusion (IRI), leukopenia and lymphopenia.

[0043] In one aspect, administration of an anti-CD38 antibody causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% depletion of the red blood cell population.

[0044] In one aspect, administration of an anti-CD38 antibody causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% platelet population depletion.

[0045] In one aspect, the hematologic malignancy is selected from the group consisting of multiple myeloma, chronic lymphoblastic leukemia, chronic lymphocytic leukemia, plasma cell leukemia, acute myeloid leukemia, chronic myeloid leukemia, B-cell lymphoma, and Burkitt's lymphoma.

[0046] In one aspect, the hematologic malignancy is multiple myeloma.

[0047] In one aspect, the VH chain region contains an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 10 Accordingly, the VH chain region may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 9, and the VL chain region may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 10. Accordingly , the VH chain region may comprise an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 9, and the VL chain region may comprise an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 10. Accordingly, the region the VH chain region may contain an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 10. Accordingly, the VH chain region may comprise an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 10. Accordingly, the VH chain region may comprise an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 10.

[0048] Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 80% sequence identity to SEQ ID NO: 9, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 80% sequence identity to SEQ ID NO: 10 Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 85% sequence identity to SEQ ID NO: 9. and the VL chain may comprise the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 85% sequence identity to SEQ ID NO: 10. Accordingly , the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 90% sequence identity to SEQ ID NO: 9, and the chain VL may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 90% sequence identity to SEQ ID NO: 10. Accordingly, the chain VH may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 95% sequence identity to SEQ ID NO: 9, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 95% sequence identity with SEQ ID NO: 10. Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 97% sequence identity with SEQ ID NO: 9, and the VL chain may contain the sequences CDRs defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 97% sequence identity with SEQ ID NO: 10. Accordingly, the VH chain may contain sequences CDRs defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 99% sequence identity to SEQ ID NO: 9, and the VL chain may contain CDR sequences, defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 99% sequence identity with SEQ ID NO: 10.

[0049] In one aspect, the VH chain region has the amino acid sequence SEQ ID NO: 9 or a variant thereof with up to three amino acid substitutions, and the VL chain region has the amino acid sequence SEQ ID NO: 10 or a variant thereof with up to three amino acid substitutions.

[0050] In one aspect, the VH chain region has the amino acid sequence of SEQ ID NO: 9, and the VL chain region has the amino acid sequence of SEQ ID NO: 10.

[0051] In one aspect, the VH chain region contains an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 12 Accordingly, the VH chain may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 11, and the VL chain region may contain an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 12.

[0052] Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 80% sequence identity to SEQ ID NO: 11, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 80% sequence identity to SEQ ID NO: 12 Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 85% sequence identity to SEQ ID NO: 11. and the VL chain may comprise the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 85% sequence identity to SEQ ID NO: 12. Accordingly , the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 90% sequence identity to SEQ ID NO: 11, and the chain VL may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 90% sequence identity to SEQ ID NO: 12. Accordingly, the chain VH may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 95% sequence identity to SEQ ID NO: 11, and the VL strand may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 95% sequence identity with SEQ ID NO: 12. Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 97% sequence identity with SEQ ID NO: 11, and the VL chain may contain the sequences CDRs defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 97% sequence identity with SEQ ID NO: 12. Accordingly, the VH chain may contain the sequences CDRs defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 99% sequence identity to SEQ ID NO: 11, and the VL chain may contain CDR sequences, defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 99% sequence identity with SEQ ID NO: 12.

[0053] In one aspect, the VH chain region has the amino acid sequence SEQ ID NO: 11 or a variant thereof with up to three amino acid substitutions, and the VL chain region has the amino acid sequence SEQ ID NO: 12 or a variant thereof with up to three amino acid substitutions.

[0054] In one aspect, the anti-CD38 antibody comprises the heavy chain amino acid sequence of SEQ ID NO: 11 and the light chain amino acid sequence of SEQ ID NO: 12.

[0055] In one aspect, a therapeutically effective amount is a dose of 45 to 1800 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 45 to 1200 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 45 to 600 milligrams. Accordingly, a therapeutically effective amount may be a dose of 45 to 135 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 135 to 1800 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 135 to 1200 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 135 to 600 milligrams. Accordingly, a therapeutically effective amount may be a dose of from 600 to 1800 milligrams. Accordingly, a therapeutically effective amount may be a dose of 600 to 1200 milligrams. Accordingly, a therapeutically effective amount may be a dose of 1,200 to 1,800 milligrams.

[0056] In one aspect, the human anti-CD38 antibody is administered in the form of a pharmaceutically acceptable composition. Accordingly, the pharmaceutically acceptable composition may be suitable for subcutaneous administration.

[0057] In another aspect, the present invention provides a unit dosage form containing an isolated antibody that contains a heavy chain variable region amino acid sequence having at least 80% identity with SEQ ID NO: 9 and a light chain variable region amino acid sequence having at least 80% sequence identity to SEQ ID NO: 10, wherein the isolated antibody binds to CD38, and the unit dosage form is prepared for subcutaneous administration of the antibody at a dosage of 45 to 1800 milligrams. Accordingly, the VH chain region may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 9, and the VL chain region may comprise an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 10. Accordingly, the VH chain region may comprise an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 9, and the VL chain region may comprise an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 10. Accordingly, the chain region The VH chain may comprise an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 9, and the VL chain region may comprise an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 10. Accordingly, the VH chain region may contain an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 10. Accordingly, the VH chain region may contain an amino acid sequence a sequence having at least 99% sequence identity to SEQ ID NO: 9, and the VL chain region contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 10.

[0058] Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 85% sequence identity to SEQ ID NO: 9, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 85% sequence identity to SEQ ID NO: 10 Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 85% sequence identity to SEQ ID NO: 9. and the VL chain may comprise the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 85% sequence identity to SEQ ID NO: 10. Accordingly , the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 90% sequence identity to SEQ ID NO: 9, and the chain VL may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 90% sequence identity to SEQ ID NO: 10. Accordingly, the chain VH may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 95% sequence identity to SEQ ID NO: 9, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 95% sequence identity with SEQ ID NO: 10. Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 97% sequence identity with SEQ ID NO: 9, and the VL chain may contain the sequences CDRs defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 97% sequence identity with SEQ ID NO: 10. Accordingly, the VH chain may contain the sequences CDRs defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 99% sequence identity to SEQ ID NO: 9, and the VL chain may contain CDR sequences, defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 99% sequence identity with SEQ ID NO: 10.

[0059] Accordingly, the present invention may provide a dosage unit form containing an isolated antibody that contains the heavy chain variable region amino acid sequence of SEQ ID NO: 9 or a variant thereof having up to three amino acid substitutions and the light chain variable region amino acid sequence of SEQ ID NO: 10 or a variant thereof having up to three amino acid substitutions, wherein the isolated antibody binds to CD38, wherein the unit dosage form is prepared for subcutaneous administration of the antibody at a dosage of 45 to 1800 milligrams.

[0060] In another aspect, the present invention provides a unit dosage form containing an isolated antibody that contains the heavy chain variable region amino acid sequence of SEQ ID NO: 9 and the light chain variable region amino acid sequence of SEQ ID NO: 10, wherein the isolated antibody binds to CD38 and does not bind significantly to human red blood cells, and the unit dosage form is prepared for subcutaneous administration of the antibody at a dosage of 45 to 1800 milligrams.

[0061] In one aspect, the heavy chain comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 11, and the light chain comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 12. Accordingly, , the VH chain may contain an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may comprise an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence sequence having at least 95% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 11, and the VL chain region contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain CDR sequences defined in SEQ ID NO: 3 , SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 80% sequence identity to SEQ ID NO: 11, and the VL chain may contain CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 80% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 85% sequence identity to SEQ ID NO: 11, and the VL chain may contain CDR sequences defined in SEQ ID NO: 6, SEQ ID NO : 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 85% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain CDR sequences defined in SEQ ID NO: 3, SEQ ID NO : 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 90% sequence identity to SEQ ID NO: 11, and the VL chain may contain CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 90% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 95% sequence identity to SEQ ID NO: 11, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 95% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 97% sequence identity to SEQ ID NO: 11, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO : 8, and the remainder of the VL sequence may have at least 97% sequence identity to SEQ ID NO: 12. Accordingly, the VH chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO : 5, and the remainder of the sequence may have at least 99% sequence identity to SEQ ID NO: 11, and the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8 and the remainder of the VL sequence may have at least 99% sequence identity to SEQ ID NO: 12.

[0062] Accordingly, the heavy chain may comprise the amino acid sequence SEQ ID NO: 11 or a variant thereof having up to three amino acid substitutions, and the light chain may contain the amino acid sequence SEQ ID NO: 12 having up to three amino acid substitutions.

[0063] In one aspect, the heavy chain may comprise the amino acid sequence of SEQ ID NO: 11 and the light chain may comprise the amino acid sequence of SEQ ID NO: 12.

[0064] In one aspect, the unit dosage form is prepared for subcutaneous administration of an antibody in the treatment of a hematologic malignancy selected from the group consisting of multiple myeloma, chronic lymphoblastic leukemia, chronic lymphocytic leukemia, plasma cell leukemia, acute myeloid leukemia, chronic myeloid leukemia, B -cell lymphoma and Burkitt's lymphoma.

[0065] In one aspect, the hematologic malignancy is multiple myeloma.

[0066] In one aspect, the anti-CD38 antibody does not cause hemolytic anemia or thrombocytopenia.

[0067] In one aspect, the anti-CD38 antibody causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% depletion red blood cell populations.

[0068] In one aspect, the anti-CD38 antibody causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% depletion platelet populations.

[0069] In one aspect, a human anti-CD38 antibody is provided for use in therapy, wherein the antibody does not cause significant levels of red blood cell depletion and/or platelet depletion upon administration. Accordingly, the human anti-CD38 antibody can be administered subcutaneously. Accordingly, the antibody can be administered at a dose of 45 to 1800 milligrams.

[0070] In one aspect, a human anti-CD38 antibody is provided for use in therapy, wherein the antibody does not cause significant levels of red blood cell depletion and/or platelet depletion upon administration, and the human anti-CD38 antibody is administered subcutaneously at a dose of 45 to 1800 milligrams. Accordingly, a human anti-CD38 antibody that does not cause a significant level of red blood cell depletion and/or platelet depletion upon administration may be an anti-CD38 antibody as defined herein.

[0071] In one aspect, a unit dosage form is provided containing an isolated antibody that does not cause a significant level of red blood cell depletion and/or platelet depletion upon administration, wherein the isolated antibody binds to CD38 and does not bind to human red blood cells, and the unit dosage form is formulated ​for subcutaneous administration of the antibody in a dose of 45 to 1800 milligrams.

[0072] In one aspect, a human anti-CD38 antibody, as defined herein, is provided for use in therapy, wherein the human anti-CD38 antibody is formulated for subcutaneous administration. Accordingly, the human anti-CD38 antibody is administered subcutaneously.

[0073] In one aspect, a human anti-CD38 antibody, as defined herein, is provided for use in treating a disease in which binding to CD38 is indicated, wherein the human anti-CD38 antibody is formulated for subcutaneous administration. Accordingly, the human anti-CD38 antibody is administered subcutaneously.

[0074] In many aspects, the dosage of the administered anti-CD38 antibody as described herein is a once-weekly dosage.

[0075] In one aspect, a human anti-CD38 antibody, as defined herein, is provided for use in therapy, wherein the human anti-CD38 antibody is formulated for subcutaneous administration. Accordingly, the human anti-CD38 antibody is administered subcutaneously.

[0076] Accordingly, the human anti-CD38 antibody can be administered at a dose ranging from 45 to 1800 milligrams of antibody. Accordingly, the human anti-CD38 antibody can be formulated for subcutaneous administration. Accordingly, the human anti-CD38 antibody can be formulated for subcutaneous administration and administered at a dose ranging from 45 to 1800 milligrams of antibody.

[0077] In one aspect, a human anti-CD38 antibody, as defined herein, is provided for use in the treatment of cancer. Accordingly, the cancer may be a hematological malignancy.

[0078] In one aspect, a human anti-CD38 antibody, as defined herein, is provided for use in the treatment of a hematologic malignancy, wherein the human anti-CD38 antibody is formulated for subcutaneous administration. Accordingly, the human anti-CD38 antibody can be administered subcutaneously.

[0079] Accordingly, the human anti-CD38 antibody can be administered at a dose ranging from 45 to 1800 milligrams of antibody. Accordingly, the human anti-CD38 antibody can be formulated for subcutaneous administration. Accordingly, the human anti-CD38 antibody can be formulated for subcutaneous administration and administered at a dose ranging from 45 to 1800 milligrams of antibody.

[0080] In one aspect, a human anti-CD38 antibody, as defined herein, is provided for use in the treatment of a hematologic malignancy, wherein the human anti-CD38 antibody is formulated for subcutaneous administration, and the human anti-CD38 antibody is administered at a dose ranging from 45 to 1800 milligrams of antibody. Accordingly, the human anti-CD38 antibody can be administered subcutaneously.

[0081] Accordingly, the hematologic malignancy may be multiple myeloma, chronic lymphoblastic leukemia, chronic lymphocytic leukemia, plasma cell leukemia, acute myeloid leukemia, chronic myeloid leukemia, B-cell lymphoma, or Burkitt's lymphoma. Accordingly, the hematologic malignancy may be multiple myeloma.

[0082] In one aspect, a human anti-CD38 antibody, as defined herein, is provided for use in treating an autoimmune disease.

[0083] In one aspect, a human anti-CD38 antibody, as defined herein, is provided for use in treating an autoimmune disease, wherein the human anti-CD38 antibody is formulated for subcutaneous administration. Accordingly, the human anti-CD38 antibody can be administered subcutaneously.

[0084] Accordingly, the human anti-CD38 antibody can be administered at a dose ranging from 45 to 1800 milligrams of antibody. Accordingly, the human anti-CD38 antibody can be formulated for subcutaneous administration. Accordingly, the human anti-CD38 antibody can be formulated for subcutaneous administration and administered at a dose ranging from 45 to 1800 milligrams of antibody.

[0085] Accordingly, the autoimmune disease may be systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), ulcerative colitis, systemic light chain amyloidosis, or graft-versus-host disease.

[0086] In one aspect, a pharmaceutical composition is provided comprising an isolated human anti-CD38 antibody, as defined herein.

[0087] In one aspect, a pharmaceutical composition is provided comprising a unit dosage form of the present invention.

[0088] In one aspect, a pharmaceutical composition of the present invention is provided for use in therapy.

[0089] In one aspect, a pharmaceutical composition of the present invention is provided for use in the treatment of a disease in which binding to CD38 is indicated.

[0090] In one aspect, a pharmaceutical composition of the present invention is provided for use in treating an autoimmune disease. Accordingly, the autoimmune disease may be systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), ulcerative colitis (UC), systemic light chain amyloidosis, or graft-versus-host disease. Accordingly, the autoimmune disease may be systemic lupus erythematosus (SLE). Accordingly, rheumatoid arthritis (RA) may be an autoimmune disease. Accordingly, an autoimmune disease may be inflammatory bowel disease (IBD). Accordingly, ulcerative colitis (UC) may be an autoimmune disease. Accordingly, an autoimmune disease may be graft-versus-host disease.

[0091] In another aspect, a pharmaceutical composition of the present invention is provided for use in the treatment of cancer. Accordingly, the cancer may be a hematologic malignancy. Accordingly, the hematologic malignancy may be multiple myeloma, chronic lymphoblastic leukemia, chronic lymphocytic leukemia, plasma cell leukemia, acute myeloid leukemia, chronic myeloid leukemia, B-cell lymphoma, or Burkitt's lymphoma. Accordingly, the hematologic malignancy may be multiple myeloma. Accordingly, the hematologic malignancy may be chronic lymphoblastic leukemia. Accordingly, the hematologic malignancy may be chronic lymphocytic leukemia. Accordingly, the hematologic malignancy may be a plasma cell leukemia. Accordingly, the hematologic malignancy may be acute myeloid leukemia. Accordingly, the hematologic malignancy may be chronic myeloid leukemia. Accordingly, the hematologic malignancy may be a B-cell lymphoma. Accordingly, the hematologic malignancy may be Burkitt's lymphoma.

[0092] In one aspect, the use of an isolated human CD38 antibody, as defined herein, is provided for the production of a medicament for the treatment of a disease.

[0093] In another aspect, there is provided the use of a unit dosage form of the present invention for the manufacture of a medicament for treating a disease.

[0094] Accordingly, the disease may be one in which binding to CD38 is indicated.

[0095] Accordingly, the disease may be an autoimmune disease such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), ulcerative colitis (UC), systemic light chain amyloidosis, or graft-versus-host disease.

[0096] Accordingly, the disease may be cancer. Accordingly, the cancer may be a hematologic malignancy such as multiple myeloma, chronic lymphoblastic leukemia, chronic lymphocytic leukemia, plasma cell leukemia, acute myeloid leukemia, chronic myeloid leukemia, B-cell lymphoma, or Burkitt's lymphoma.

[0097] Accordingly, the drug can be formulated for subcutaneous administration.

[0098] Accordingly, the drug can be formulated to provide a dose of from 45 to 1800 milligrams of antibody.

[0099] Accordingly, the drug can be formulated for subcutaneous administration at a dose of 45 to 1800 milligrams of antibody.

[00100] These and other embodiments, features, and potential advantages will become apparent with reference to the following description and drawings.

Brief description of graphic materials

[00101] The subject matter and features of the invention may be better understood by reference to the drawings described below, in which:

[00102] In FIG. Figure 1 shows a table of antibodies used for flow cytometric analysis in PD research.

[00103] In FIG. Figure 2 shows the PK data of the subcutaneous dose groups. Anti-drug antibodies (ADA) were detected using a validated qualitative electrochemiluminescence (ECL) assay. The incidence increased over time and affected PK when it reached a certain threshold titer of about 1000 (~log(7)).

[00104] In FIG. Figure 3 shows FC data from the cynomolgus monkey (macaque) and the AB79 model. Panels A and B show raw PK data from 8 monkey studies, panel A for the first 7 days after the first dose and panel B for the entire follow-up period. Doses were color coded, but SC data were not reported (Figure 2). Panel C shows the final structure of the PK model, including target-mediated drug distribution (TMDD), indicated by the blue box. V _C denotes the volume of the central compartment in which concentrations of AB79 are observed (labeled Conc). V _P denotes the volume of the peripheral compartment. R _total represents the antibody-bound and non-antibody-bound CD38 receptor compartment. K _SYN and K _DEG denote the rate constants for receptor production and degradation, and K _INT the rate constant for internalization (complex rate constant for elimination). K _SS is the steady state constant, defined as K _SS = (K _OFF +K _INT ) / K _ON , where K _OFF is the dissociation and K _ON is the binding rate constant. DF panels show overlaps of linear two-compartment model predictions (median, 95% prediction interval) without the TMDD component and observed data for the 3 lowest doses (Study 8). Note the different time frames between panels D , E and F.

[00105] In FIG. Figure 4 shows the effect of AB79 treatment on red blood cells two days post-dose in Study 7 and total lymphocyte counts on the first day post-dose.

[00106] In FIG. Figure 5 shows the effects of ADA in a 13-week toxicology study. The assessment refers to the final population PK model (Figure 1, Table 4). The following goodness-of-fit (GOF) plots are presented, stratified by dose and route of administration (Keizer et al. (2013) CPT Pharmacometrics Syst. Pharmacol. 2:e50): (1) conditional weighted residuals (CWRES) versus time; (2) observed concentration versus population model prediction; (3) CWRES versus population model prediction; and (4) observed concentration compared to individual model prediction.

[00107] In FIG. Figure 6 shows GOF plots for the final population PK model, stratified by dose and route of administration (IV - red, SC - blue).

[00108] In FIG. Figure 7 shows a comparison of CD38 expression on the surface of human and monkey NK, B and T cells. Flow cytometry measurements were standardized and signals are expressed in molecules of equivalent soluble fluorescence (MOEF). Human and monkey blood lymphocytes bind similar levels of AB79. Direct comparison of CD38 expression levels on monkey NK cells (CD3-, CD159a+), B cells (CD3-, CD20+) and T cells (CD3+) and human NK cells (CD3-, CD16/CD56+) , B cells (CD3−, CD19+) and T cells (CD3+) were assessed using flow cytometry. Median fluorescence intensity (MFI) for AB79 staining for each cell population was converted to MOEF units using a standard curve generated using Rainbow Beads (Spherotech; Lake Forest, IL). Data shown are from 3 individuals of each species and show MOEF ± SD for each cell type. There are differences in CD38 expression between blood lymphocytes with higher levels of AB79 binding (MOEF) on NK cells > B cells > T cells. The pattern of AB79 binding is similar in monkey blood cells, but the level of AB79 binding/CD38 expression is lower.

[00109] In FIG. Figure 8 shows inter- and intra-individual variability in T cell, B cell and NK cell count data in placebo-treated animals.

[00110] In FIG. Figure 9 shows pre-injection NK, B and T cell counts (cells per µL) stratified by study (top row) or sex (bottom row).

[00111] In FIG. 10 shows AB79-dependent depletion of NK cells, B cells and T cells. The graphs presented focus on changes that occurred during the first 7 days after treatment with the first dose of AB79. Thus, it was possible to combine data from single-dose and multiple-dose studies with a weekly or biweekly dosing schedule. AC plots show individual minimum cell counts (i.e., maximum effect on PD), individual cell counts 7 days after the first dose, as well as average cell depletion profiles per dose and NK cell PK-PD model structure, respectively. EF plots show the same information for B cells, and GI plots show the same information for T cells.

[00112] In FIG. Figure 11 shows simulated human PK profiles and NK cell, B cell, and T cell depletion profiles for AB79. Based on scaled monkey PK and PK-PD models, 5 PK and cell removal profiles were simulated for single intravenous and subcutaneous doses (0.0003 to 1 mg/kg). The graphs on the left show data after intravenous administration, and the graphs on the right after subcutaneous administration. The first row of graphs displays the FC profiles. The lower limit of quantitation (LLOQ) of 0.05 μg/mL is indicated by the horizontal dotted line. The lowest dose PK was completely obscured by noise, and only at doses of 0.03 mg/kg did PK reach levels above the LLOQ.

[00113] In FIG. 12 shows the design of a single escalating dose study of AB79 in healthy volunteers (toxicity study). A total of 6 IV and 4 SC cohorts of 74 subjects were randomized to receive a single dose of AB79. Extended blinded analysis of safety, PK and PD data was performed for each cohort before dose escalation. Stopping criteria included depletion of the target cell population to avoid potential immunosuppression in healthy volunteers. Each subject was observed for 92 days after drug administration.

[00114] OnFig. 13showing GOF plots for PK-PD models stratified by path administration (iv - red; s/c - blue). A) NK cells. B) B cells. C) T cells.

[00115] In FIG. 14 shows that AB79 mediates cell population depletion through antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Comparison of CD38 receptor abundance and susceptibility to ADCC and CDC in human B cell lines. Cell lines with increased expression of CD38 were more susceptible to ADCC. No ADCC was observed in a human lymphoblast cell line that did not express CD38 (MV-4-11) or in a Chinese hamster ovary cell line transfected with CD157, a molecule related to CD38 (data not shown). EC ₅₀ -50% effective concentration; n/v - not performed; SD - standard deviation.

[00116] In FIG. 15 shows that AB79 mediates the depletion of lymphocyte populations in monkeys. AB79 dose-dependently depleted blood NK cell > B cell > T cell populations in female cynomolgus monkeys (n=4/dose group) following a single intravenous dose of AB79, as quantified using Flow-Count ^™ fluorospheres (Beckman-Coulter) with using flow cytometry. Samples were collected during pre-treatment (week -1), day 1: pre-dose, post-dose at 15, 30 minutes, 1, 4, 8, 24, 48, 96 and 168 hours, at 10, 15, 22, 29, 36, 43, 50 and 57 days. For clarity, only 2 weeks of data are shown. Average cell counts were calculated at each time point and used to calculate % of baseline counts.

[00117] In FIG. 16 shows that the secondary response to human tetanus toxoid (TTd) is reduced by treatment with AB79. CB17/SCID mice were treated with anti-asialized GM1 antibody to kill NK cells and then injected with 25 x 10 ⁶ human peripheral blood lymphocytes. After 7–10 days, serum samples were collected to assess human Ig, and Ig levels were the basis for randomization. Mice were given TTd to induce a response and treated with the indicated antibodies twice a week for 10 days. 3 days after the last treatment, serum was collected and analyzed for antibodies to TTd. AB79 dose-dependently suppressed the secondary response to TTd. AB79 reduced the response to the same extent as Rituxan (Rtx) (isotype (Iso), Rtx and AB79, all at 10 mg/kg).

[00118] In FIG. 17 shows that AB79 does not induce cytokines. AB79 (soluble) did not increase IL-6 levels in PBMCs collected from 4 different subjects after 24 hours of incubation compared to the IgG1 isotype control PHA (positive control) increased cytokine levels in all subjects, demonstrating that the cells have the ability to produce IL - 6. Similar results were obtained when PBMCs were stimulated for 48 hours and when tested with IL-2, IL-4, IL-10, GM-CSF, IFNγ and TNFα (data not shown).

[00119] In FIG. 18A is a diagram of the dry binding, wet binding and solute experiment shown in FIG. 18B (modified from Stebbings et al. (2007) J. Immunol. 179: 3325-3331).

[00120] In FIG. 18B shows that AB79 does not have agonist activity. AB79 was highly concentrated when added to wells in solution to evaporate liquid (dry binding) compared to AB79 that was bound to wells in solution (wet binding) or added directly to PBMC (soluble). AB79 did not stimulate IL-6 or IL-2, IL-4, IL-8, IL-10, GM-CSF, IFNγ, or TNFα in any of the conditions tested at 24 hours. IL-8 was constitutively produced by PBMCs and was not altered by any treatment (data not shown).

[00121] Figure 19 shows an assessment of AB79 binding to cynomolgus CD45+ lymphocytes. AB79 binding to CD45+ lymphocytes in unlysed cynomolgus macaque whole blood. CD45+ lymphocytes are recorded and then AB79 binding (black histogram) or isotype control binding (red histogram) is assessed. AB79 binding was detected in a subset of lymphocytes, as shown in the fraction of cells to the right of the red dotted line. Almost no binding of the isotype control to lymphocytes was observed.

[00122] In FIG. Figure 20 shows the mean observed predose Cmax and trough levels (ng/mL) (cycle 1 and cycle 2). In FIG. 20A shows the Cmax (ng/ml) of Ab79 and FIG. 20B shows the concentration of Ab79 (ng/ml).

[00123] In FIG. 21 shows that subcutaneously administered Ab79 reduces blood plasmablast levels in a dose-dependent manner.

[00124] In FIG. 22 shows that subcutaneously administered Ab79 reduces plasmablast levels in bone marrow aspirates in a dose-dependent manner.

[00125] In FIG. 23 shows that subcutaneously administered Ab79 reduces plasma cell levels in bone marrow aspirates in a dose-dependent manner.

[00126] In FIG. 24 shows NK cell levels in the peripheral blood of healthy subjects following a single SC administration of AB79. S/c - subcutaneous.

[00127] In FIG. 25 shows the levels of plasmablasts, monocytes, B, T and NK cells in the peripheral blood of healthy subjects after a single injection of placebo control, 0.1, 0.3 or 0.6 mg/kg-1 SC AB79. Absolute number of monocytes (cells/μl), …….. NK cells (cells/μl), total T-lymphocytes (cells/μl), - - - - B-cells (cells/µl), plasmablast cells (cells/μl). Centered curves represent the median value. NK - natural (cell) killer; s/c - subcutaneously.

[00128] In FIG. 26 shows the binding of AB79 and daratumumab to human red blood cells (median individual donor fluorescence). Peripheral blood of four healthy volunteers was incubated with biotin-streptavidin-BV421 AB79 (0, 0.1, 10, 100 μg/ml) or biotin-streptavidin-BV421 daratumumab (0, 0.1, 1, 10, 100 μg/ml) for 3 hours at room temperature with gentle shaking in the presence or absence of unlabeled AB79 (500 μg/ml) or unlabeled daratumumab (500 μg/ml). Explanation: AB79-biotin-strep-BV421; cold AB79 and AB79-biotin-strep-BV421; daratumumab-biotin-strep-BV421; cold daratumumab and daratumumab-biotin-strep-BV421.

Detailed description of the invention

[00129] The present invention provides methods for treating diseases associated with CD-38 by subcutaneously administering anti-CD38 antibodies.

[00130] In the vasculature of patients with active disease, approximately 36 times more CD38 molecules are expressed on red blood cells than on myeloma cells. Thus, for example, off-target expression of CD38 may be required before unbound antibody can pass into the bone marrow and bind CD38 expressed on myeloma cells. This may explain why other anti-CD38 antibodies known in the art, such as daratumumab and isatuximab, which bind tightly to red blood cells and platelets, require high systemic doses to be effective.

[00131] AB79, daratumumab, isatuximab and MOR202 are IgG1s that primarily kill tumors through antibody-dependent cellular cytotoxicity (ADCC). This mechanism requires effector cells such as NK cells to bind antibodies to target cells and form a lytic synapse for targeted secretion of cytotoxic agents. The frequency of these effector cells in the blood is several orders of magnitude lower than that of erythrocytes and platelets. For example, the ratio of red blood cells to NK cells in the blood is 20,000:1. Consequently, the effector activity of daratumumab, isatuximab and MOR202 is diverted from tumors because effector cells are primarily bound by those anti-CD-38 antibodies that are associated with red blood cells and platelets, preventing the formation of a lytic synapse with tumors, resulting in low ADCC efficacy.

[00132] Treatment of patients with antibodies to CD38, which bind to red blood cells and platelets, can lead to life-threatening side effects. For example, in one study, treatment of relapsed or refractory multiple myeloma with MOR202 resulted in several serious treatment-related adverse events or TEAEs (see, e.g., Raab et al. (2015) Blood 126: 3035). The most common TEAEs of any grade were anemia (15 patients, 34%), fatigue (14 patients, 32%), infusion-related reactions (IRR) and leukopenia (13 patients, 30% each), lymphopenia and nausea (11 patients, 25 each). % every). Grade ≥3 TEAEs were reported in 28 patients (64%); the most common were lymphopenia (8 patients, 18%), leukopenia (5 patients, 11%), and hypertension (4 patients, 9%). RSIs occurred mainly during the first administration; all were grades 1-2, with the exception of one patient (grade 3). Infections were frequently reported (26 patients, 59%), but were considered unrelated to treatment in most cases. MOR202 has been used clinically only as an intravenous infusion.

[00133] Other Morphosys antibodies are known that target CD38 (see, eg , WO 2006/125640, which discloses four human antibodies: MOR03077, MOR03079, MOR03080 and MOR03100, and two mouse antibodies: OKT10 and IB4). These prior art antibodies are inferior to antibodies for use in this invention (eg, AB79) for a number of reasons. MOR03080 binds to human CD38 and cynomolgus CD38, but with low affinity to human CD38 (Biacore K _D =27.5 nm). OKT10 binds to human CD38 and cynomolgus CD38, but with low/moderate affinity to human CD38 (Biacore K _D =8.28 nm). MOR03079 binds to human CD38 with high affinity (Biacore K _D =2.4 nm) but does not bind to cynomolgus CD38. MOR03100 and MOR03077 bind to human CD38 with moderate to low affinity (Biacore K _D =10 nm and 56 nm, respectively). In comparison, antibodies for use in the present invention (eg, AB79) bind to human CD38 and cynomolgus monkey with high affinity to human CD38. (Biacore K _D =5.4 nm). Moreover, prior art antibodies have low activity of both ADCC and CDC.

[00134] The advantage of more efficient ADCC is the ability to deliver the therapeutic CD38 antibody in a small volume injection. If an antibody for use in this invention (eg, AB79) is formulated at a concentration of 100 mg/ml, the effective dose for an 80 kg patient with myeloma can be administered as a single subcutaneous injection of <1.0 ml. In contrast, an effective dose of daratumumab or isatuximab administered to this patient in a comparable form (i.e., 100 mg/mL) would require administration of 12.8 mL or 8–16 mL, respectively.

[00135] The methods and dosage units of anti-CD38 antibodies provided herein provide subcutaneous administration of therapeutically effective doses of anti-CD38 antibodies, thereby providing unexpected benefits and preventing the side effects, inconveniences and costs associated with the administration of high doses of systemic anti-CD38 antibody therapies.

[00136] The present invention provides methods and unit dosage forms for subcutaneously administering a therapeutically effective amount of an isolated anti-CD38 antibody to a patient in need thereof for the treatment of diseases in which binding to CD38 is indicated, including hematologic malignancies. In some embodiments, the subcutaneous antibody comprises a heavy chain variable region comprising SEQ ID NO: 9 (or a sequence with at least 80%, 85%, 90%, 95%, 97%, or 99% sequence identity) and a variable region light chain containing SEQ ID NO: 10 (or sequence identity of at least 80%, 85%, 90%, 95%, 97% or 99%). The anti-CD38 antibody provided herein is capable of being therapeutically effective when administered subcutaneously.

[00137] Another advantage of the anti-CD38 antibodies of this invention is that, unlike some other anti-CD38 antibodies used clinically, the anti-CD38 antibodies of this invention (e.g., AB79) are capable of binding to CD38 of the cynomolgus monkey (macaque). , providing a useful animal model for preclinical evaluation of dosage, toxicity and efficacy, etc.

[00138] Another advantage of the anti-CD38 antibodies of this invention is that they can be used to screen for other antibodies that compete for binding to CD-38 with the same epitope and may be useful in the methods and dosage unit forms of this invention .

[00139] Unless otherwise defined herein, scientific and technical terms used in connection with this invention have the meanings commonly accepted by those skilled in the art. The meaning and scope of the terms must be clear. However, in the event of any implied ambiguity, the definitions given herein take precedence over any dictionary or external definition. In addition, unless the context otherwise requires, terms in the singular include the plural and terms in the plural include the singular. The term “or” includes “and/or” unless otherwise indicated. In addition, the use of the term “includes,” “includes,” or “inclusive” is non-limiting. The terms “element” and “component” cover both elements and components that contain a single unit and elements and components that contain more than one subunit, unless specifically stated otherwise.

[00140] In general, the nomenclature used in connection with cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization, and methods for their preparation described herein are well known and widely used. used in this field of technology. The methods and techniques of this invention are generally carried out in accordance with conventional methods well known in the art and described in the various general and more specific references cited and discussed herein unless otherwise indicated. Enzymatic reaction and purification procedures are carried out in accordance with the manufacturer's instructions, as is generally used in the art, as described herein. The nomenclatures used in connection with laboratory procedures and methods of analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are well known and widely used in the art. Standard methods are used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, delivery and patient treatment.

[00141] All headings and section designations are used for clarity and reference purposes only and should not be construed as limiting in any way. For example, those skilled in the art will appreciate the usefulness of combining the various aspects herein from the various headings and sections consistent with the spirit and scope of the invention described herein.

[00142] Certain terms are defined below for easier understanding of the present invention.

[00143] The terms “human CD38” and “human CD38 antigen” refer to the amino acid sequence of SEQ ID NO: 1 or a functional fraction thereof, such as an epitope, as defined herein (Table 1). In general, CD38 has a short intracytoplasmic tail, a transmembrane domain, and an extracellular domain. The terms “cynomolgus CD38” and “cynomolgus CD38 antigen” refer to the amino acid sequence of SEQ ID NO: 2, which is 92% identical to the amino acid sequence of human CD38 (Table 1). Synonyms for CD38 include cyclic ADP-ribose hydrolase; cyclic ADP ribose hydrolase 1; ADP-ribosyl cyclase; ADP-ribosyl cyclase 1; cADP hydrolase 1; CD38-rs1; I-19; NIM-R5 antigen; 2'-phospho-cyclic-ADP-ribose transferase; 2'-phospho-ADP-ribosyl cyclase; 2'-phospho-cyclic-ADP-ribose transferase; 2'-phospho-ADP-ribosyl cyclase; T10.

Table 1. Amino acid sequence of human and cynomolgus CD38

View Amino acid sequence
12345678901234567890123456789012345678901234567890 SEQ ID NO human CD38 MANCEFSPVSGDKPCCRLSRRAQLCLGVSILVLILVVVLAVVVPRWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRHVDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDLTWCGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRFA EAACDVVHVMLNGSRSKIFDKNSTFGSVEVHNLQPEKVQTLEAWVIHGGREDSRDLCQDPTIKELESIISKRNIQFSCKNIYRPDKFLQCVKNPEDSSCTSEI 1 Cynomolgus CD38 MANCEFSPVSGDKPCCRLSRRAQVCLGVCLLVLLILVVVVAVVLPRWRQQWSGSGTTSRFPETVLARCVKYTEVHPEMRHVDCQSVWDAFKGAFISKYPCNITEEDYQPLVKLGTQTVPCNKTLLWSRIKDLAHQFTQVQRDMFTLEDMLLGYLADDLTWCGEFNTFEINYQSCPDWRKDCSNNPVSVFWKTVS RRFAETACGVVHVMLNGSRSKIFDKNSTFGSVEVHNLQPEKVQALEAWVIHGGREDSRDLCQDPTIKELESIISKRNIRFFCKNIYRPDKFLQCVKNPEDSSCLSGI 2 Human CD157 maaqgcaasrllqlllqlllllllaaggararwrgegtsahlrdiflgrcaeyrallspeqrnknctaiweafkvaldkdpcsvlpsdydlfinlsrhsiprdkslfwenshllvnsfadntrrfmplsdvlygrvadflswcrqkndsgldyqscptsedcennpvdsfwkrasiqyskdssgvihvmlngs eptgaypikgffadyeipnlqkekitrieiwvmheiggpnvescgegsmkvlekrlkdmgfqyscindyrpvkllqcvdhsthpdcalksaaaatqrkapslyteqragliiplflvlasrtql 13

[00144] The terms “therapeutically effective amount” and “therapeutically effective dose” refer to an amount of therapy that is sufficient to reduce or alleviate the severity and/or duration of a disorder or one or more symptoms thereof; preventing the development of the disorder; causing regression of the disorder; preventing relapse, development, occurrence or progression of one or more symptoms associated with the disorder; or enhancing or improving the prophylactic or therapeutic effect(s) of another therapy (e.g., a prophylactic or therapeutic agent) in doses and for periods of time necessary to achieve the desired therapeutic result. The therapeutically effective amount may vary depending on factors such as the disease state, age, sex and weight of the individual, as well as the ability of the drugs to produce the desired response in the individual. A therapeutically effective amount of an antibody is an amount such that any toxic or harmful effects of the antibody or antibody portion are outweighed by the therapeutically beneficial effects. A therapeutically effective amount of an antibody for tumor therapy can be measured by its ability to stabilize disease progression. The ability of a compound to inhibit cancer can be assessed in an animal model system predictive of efficacy against human tumors.

[00145] The terms “patient” and “subject” include both humans and other animals, particularly mammals. Thus, the compositions, dosages and methods disclosed herein are applicable to both human and veterinary medicine. In one embodiment, the patient is a mammal, such as a human.

[00146] The term “disease for which binding to CD38 is indicated” means a disease in which binding of a binding partner (eg, an anti-CD38 antibody of the invention) to CD38 provides a prophylactic or therapeutic effect, including improvement of one or more symptoms of the disease. Such binding may lead to blocking of other factors or binding partners for CD38, neutralization of CD38, ADCC, CDC, complement activation, or some other mechanism by which disease is prevented or treated. Binding factors and partners for CD38 include anti-CD38 autoantibodies, which are blocked by the anti-CD38 antibodies of the invention. Such binding may be indicated as a consequence of expression of CD38 by cells or a subset of cells, for example MM cells, whereby provision of a CD38 binding partner to a subject results in the removal, for example, lysis of these cells, for example through hemolysis or apoptosis. Such CD38 expression may be, for example, normal, overexpressed, inappropriately expressed, or a consequence of activation of CD38 relative to normal cells or relative to other cell types, either during a non-disease state or a disease state.

[00147] The term “hematologic malignancy” refers to malignancies of hematopoietic tissues and includes leukemias, lymphomas, and multiple myelomas. Non-limiting examples of conditions associated with aberrant expression of CD38 include, but are not limited to, multiple myeloma (Jackson et al . (1988) Clin. Exp. Immunol. 72: 351-356); B-cell chronic lymphocytic leukemia (B-CLL) (Dürig et al . (2002) Leukemia 16: 30-35; Morabito et al. (2001) Leukemia Res. 25: 927-932; Marinov et al . (1993) Neoplasma 40(6): 355-358 and Jelinek et al (2001) Br J Haematol 115: 854-861); acute lymphoblastic leukemia (Keyhani et al . (1999) Leukemia Res. 24: 153-159 and Marinov et al . (1993) Neoplasma 40(6): 355-358); chronic myeloid leukemia (Marinov et al . (1993) Neoplasma 40(6): 355-358); acute myeloid leukemia (Keyhani et al . (1999) Leukemia Res. 24: 153-159); chronic lymphocytic leukemia (CLL); chronic myeloid leukemia or chronic myeloid leukemia (CML); acute myeloid leukemia or acute myeloid leukemia (AML); acute lymphocytic leukemia (ALL); hairy cell leukemia (HCL); myelodysplastic syndromes (MDS); and all subtypes and stages (eg, blast phase (BP) of CML, chronic phase (CP), or accelerated phase (AP)) of these leukemias and other hematological diseases, which are determined by morphological, histochemical and immunological methods well known to those skilled in the art .

[00148] The terms "neoplasm" and "neoplastic condition" refer to a condition associated with cell proliferation characterized by loss of normal control, resulting in one or more symptoms including unregulated growth, lack of differentiation, dedifferentiation, local tissue invasion and metastasis.

[00149] The term “isolated antibody” refers to an antibody that is substantially free of other antibodies having different antigen specificities. For example, an isolated antibody that specifically binds to CD38 is substantially free of antibodies that specifically bind antigens other than CD38. However, an isolated antibody that specifically binds to an epitope, isoform, or variant of human CD38 or cynomolgus CD38 may be cross-reactive with other related antigens, for example, from other species, such as CD38 species homologs. Moreover, the isolated antibody may contain substantially no other cellular material and/or chemicals.

[00150] The terms “red blood cells,” “RBCs,” and “erythrocytes” refer to hemoglobin-containing cells in the bone marrow that carry oxygen to cells and tissues, and which carry carbon dioxide back to the respiratory system. Red blood cells are also called red cells, red blood cells, and erythroid cells.

[00151] The terms “specific binding,” “specifically binds to,” and “specific to,” when applied to the interaction of a particular antibody, protein, or peptide with an antigen, epitope, or other chemical species, mean binding that is substantially different from nonspecific interaction. Specific binding can be measured, for example, by determining the binding of a molecule compared to the binding of a control molecule, which is generally a molecule of similar structure that does not have binding activity. For example, specific binding can be determined by competition with a control molecule similar to the target. The anti-CD38 antibodies of the present invention specifically bind CD38 ligands. The terms "specific binding", "specifically binds to" and "specific to" also mean that the interaction depends on the presence of a particular structure (eg, antigenic determinant or epitope) of the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins in general. If the antibody is specific for epitope "A", the presence of a molecule containing epitope A (or free unlabeled A) in the reaction containing labeled "A" and the antibody will reduce the amount of labeled A bound to the antibody. Specific binding to a particular antigen or epitope may be exhibited, for example, by an antibody having a KD for the antigen or epitope of at least about 10 ^-4 M, at least about 10 ^-5 M, at least about 10 ^-6 M, at least at least about 10 ^-7 M, at least about 10 ^-8 M, at least about 10 ^-9 M, at least about 10 ^-10 M, at least about 10 ^-11 M, at least about 10 ^-12 M or more, where KD denotes the rate of dissociation of a particular antibody-antigen interaction. Typically, an antibody that specifically binds an antigen will have a KD that is 20-, 50-, 100-, 500-, 1000-, 5000-, 10,000-, or more times greater for the reference molecule compared to the antigen or epitope. In addition, specific binding to a particular antigen or epitope may be exhibited, for example, with an antibody having a KA or Ka for the antigen or epitope of at least 20, 50, 100, 500, 1000, 5000, 10,000, or more than times greater for an epitope compared to a control, where KA or Ka refers to the rate of association of a particular antibody-antigen interaction.

[00152] The term "over a period of time" refers to any period of time, such as minutes, hours, days, months or years. For example, over a period of time may mean at least 10 minutes, at least 15 minutes, at least 30 minutes, at least 60 minutes, at least 75 minutes, at least 90 minutes, at least 105 minutes, at least 120 minutes, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 12 hours, at least 14 hours, at least 16 hours, at least 18 hours, at least 20 hours, at least 22 hours, at least one day, at least two days , at least three days, at least 4 days, at least 5 days, at least 6 days, at least once a week, at least once a month, at least once a year or any the period of time between them. In other words, the antibody of the composition can be absorbed by the individual to whom it is administered over a period of at least 10 minutes, at least 15 minutes, at least 30 minutes, at least 60 minutes, at least 75 minutes, at least 90 minutes, at least 105 minutes, at least 120 minutes, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours , at least 9 hours, at least 10 hours, at least 12 hours, at least 14 hours, at least 16 hours, at least 18 hours, at least 20 hours, at least 22 hours, according to at least one day, at least two days, at least three days, at least 4 days, at least 5 days, at least 6 days, at least once a week, at least once a month , at least once a year or any period of time in between.

[00153] A composition that “substantially” contains a component means that the composition contains more than about 80% by weight of the component. Accordingly, the composition may contain more than about 90% by weight of the component. Accordingly, the composition may contain more than about 95% by weight of the component. Accordingly, the composition may contain more than about 97% by weight of the component. Accordingly, the composition may contain more than about 98% by weight of the component. Accordingly, the composition may contain more than about 99% by weight of the component.

[00154] The term "about" refers to a degree close in amount, degree, volume, time, etc., with minor variations of up to 10%.

[00155] The term “pharmaceutically acceptable carrier” refers to a pharmaceutically acceptable material, composition or carrier suitable for administering the compounds of this invention to mammals. Carriers include a liquid or solid filler, diluent, excipient, solvent or encapsulating material involved in carrying or transporting the subject compound from one organ or body part to another organ or body part. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the drug and not causing harm to the patient. In one embodiment, the pharmaceutically acceptable carrier is suitable for intravenous administration. In another embodiment, the pharmaceutically acceptable carrier is suitable for locoregional injection. In another embodiment, the pharmaceutically acceptable carrier is suitable for subcutaneous administration. In another embodiment, the pharmaceutically acceptable carrier is suitable for subcutaneous injection.

[00156] The term "pharmaceutical composition" refers to preparations suitable for administration to a subject and treatment of a disease. When the anti-CD38 antibodies of the present invention are administered as pharmaceutical preparations to mammals, such as humans, they can be administered "as is" or in the form of a pharmaceutical composition containing the anti-CD38 antibody in combination with a pharmaceutically acceptable carrier and/or other excipients. The pharmaceutical composition may be in the form of a unit dosage form for administering a specific dose of an anti-CD38 antibody at a specific concentration, a specific amount, or a specific volume. Pharmaceutical compositions are provided containing antibodies to CD38, either alone or in combination with prophylactic agents, therapeutic agents and/or pharmaceutically acceptable carriers. Accordingly, the pharmaceutical composition may include a unit dosage form according to this invention, either alone or in combination with prophylactic agents, therapeutic agents and/or pharmaceutically acceptable carriers. Accordingly, the pharmaceutical composition may comprise a human anti-CD38 antibody as described herein, either alone or in combination with prophylactic agents, therapeutic agents and/or pharmaceutically acceptable carriers.

[00157] Traditional antibody structural units typically contain a tetramer. Each tetramer typically consists of two identical pairs of polypeptide chains, each pair having one "light" chain (usually with a molecular weight of about 25 kDa) and one "heavy" chain (usually with a molecular weight of about 50-70 kDa). Human light chains are classified as kappa and lambda light chains. Heavy chains are classified as mu, delta, gamma, alpha, or epsilon and define the antibody isotype as IgM, IgD, IgG, IgA, and IgE, respectively. IgG has several subclasses, including but not limited to IgG1, IgG2, IgG3, and IgG4. IgM has subclasses including, but not limited to, IgM1 and IgM2. Thus, "isotype" refers to any of the subclasses of immunoglobulins, defined by the chemical and antigenic characteristics of their constant regions. The known isotypes of human immunoglobulins are IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgM1, IgM2, IgD and IgE. Therapeutic antibodies may also include isotype and/or subclass hybrids.

[00158] Each heavy chain variable region (VH) and light chain variable region (VL) (about 100-110 amino acids long) consists of three hypervariable regions called “complementarity determining regions” (CDRs) and four framework regions (FRs). ) (about 15-30 amino acids in length), located from the amino terminus to the carboxy terminus in the following order: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. "Variability" refers to the fact that CDRs vary widely in sequence among antibodies and thus define a unique antigen binding site.

[00159] The hypervariable region typically includes amino acid residues of about 24-34 amino acid residues (LCDR1; "L" is light chain), 50-56 (LCDR2) and 89-97 (LCDR3) in the light chain variable region and about 31- 35B (HCDR1; "H" denotes heavy chain), 50-65 (HCDR2) and 95-102 (HCDR3) in the heavy chain variable region (Kabat et al. (1991) Sequences Of Proteins Of Immunological Interest, ^5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD) and/or residues forming a hypervariable loop (for example, residues 26-32 (LCDR1), 50-52 (LCDR2) and 91-96 (LCDR3) in the light chain variable region and 26-32 (HCDR1), 53-55 (HCDR2) and 96-101 (HCDR3) in the heavy chain variable region (Chothia and Lesk (1987) J. Mol. Biol. 196: 901-917.

[00160] The Kabat numbering system is typically used to designate a residue in a variable domain (approximately light chain variable region residues 1-107 and heavy chain variable region residues 1-113) (eg, Kabat et al. (1991) Sequences Of Proteins Of Immunological Interest, ^5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD), with the EU numbering system used for the Fc region.

[00161] The term "immunoglobulin (Ig) domain" refers to a region of an immunoglobulin having a distinct tertiary structure. In addition to variable domains, each heavy and light chain has constant domains: heavy chain constant domains (CH); light chain constant (CL) domains and hinge domains. In the context of IgG antibodies, each IgG isotype has three CH regions. The carboxy-terminal portion of each HC and LC defines a constant region primarily responsible for effector function. Accordingly, the "CH" domains in the context of IgG are as follows: "CH1" refers to positions 118-220 according to the EU index, as in Kabat. "CH2" refers to positions 237-340 according to the EU index, as in Kabat, and "CH3" refers to positions 341-447 according to the EU index, as in Kabat.

[00162] Another type of heavy chain Ig domain is the hinge region. The term “hinge region” refers to a flexible polypeptide containing the amino acids between the first and second constant domains of an antibody. Structurally, the IgG CH1 domain ends at EU position 220, and the IgG CH2 domain begins at residue EU position 237. Thus, for IgG, the antibody hinge is defined herein to include positions 221 (D221 in IgG1) to 236 (G236 in IgG1), where the numbering corresponds to the UE index, as in Kabat. In some embodiments, such as in the context of region Fc, a bottom hinge is included, with "bottom hinge" typically referring to positions 226 or 230.

[00163] The term "Fc region" refers to a polypeptide containing the constant region of an antibody, excluding the first immunoglobulin constant domain and in some cases the hinge portion. Thus, Fc refers to the last two constant regions of the immunoglobulin domains IgA, IgD and IgG and the last three constant regions of the immunoglobulin domains IgE and IgM, as well as the flexible hinge N-terminus of these domains. For IgA and IgM, the Fc may contain a J chain. For IgG, the Fc domain includes the immunoglobulin domains Cγ2 and Cγ3 (Cγ2 and Cγ3) and the lower hinge region between Cγ1 (Cγ1) and Cγ2 (Cγ2). Although the boundaries of the Fc region may vary, the Fc region of the human IgG heavy chain is generally defined to include residues C226 or P230 at its carboxyl terminus, where the numbering corresponds to the EU index, as in Kabat. In some embodiments, as more fully described below, amino acid modifications are made to the Fc region, for example, to alter binding to one or more FcγR receptors or an FcRn receptor.

Antibodies to CD38

[00164] Accordingly, the present invention provides isolated anti-CD38 antibodies that specifically bind human and primate CD38 protein, which find use in subcutaneous administration routes and unit dosage forms. In particular, the present invention utilizes antibodies that bind to both human and primate CD38 proteins, particularly primates used in clinical trials such as cynomolgus monkeys ( Macaca fascicularis , also referred to herein as "macaques"). ").

[00165] In some embodiments, the anti-CD38 antibodies of the invention interact with CD38 at a number of amino acid residues, including K121, F135, Q139, D141, M142, E239, W241, S274, C275, K276, F284, V288, K289, N290, P291 , E292, D293, and S294 based on human sequence numbering. Accordingly, the anti-CD38 antibodies of the invention can interact with CD38 at a number of amino acid residues, including K121, F135, Q139, D141, M142, E239, W241, S274, C275, K276, F284, V288, K289, N290, P291, E292, D293 and S294 SEQ ID NO: 1 based on human sequence numbering. Accordingly, the anti-CD38 antibodies of the invention interact with CD38 at a number of amino acid residues, including K121, F135, Q139, D141, M142, E239, W241, F274, C275, K276, F284, V288, K289, N290, P291, E292, D293 and S294 SEQ ID NO: 2. It should be noted that these residues are identical in both humans and cynomolgus monkeys, except that S274 is actually F274 in cynomolgus monkeys. These residues may be an immunodominant epitope and/or trace residues of a specific antigen binding peptide.

[00166] In some embodiments, an anti-CD38 antibody for use according to the invention comprises a heavy chain comprising the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO: 3; HCDR1 AB79), ISWNGGKT (SEQ ID NO: 4; HCDR2 AB79) and ARGSLFHDSSGFYFGH ( SEQ ID NO: 5; HCDR3 AB79) or variants of these sequences containing up to three amino acid substitutions. In some embodiments, an antibody for use according to the invention contains a light chain containing the following CDR amino acid sequences: SSNIGDNY (SEQ ID NO:6; LCDR1 AB79), RDS (SEQ ID NO:7; LCDR2 AB79) and QSYDSSLSGS (SEQ ID NO:8 ; LCDR3 AB79) or variants of these sequences containing up to three amino acid substitutions. In some embodiments, an antibody for use according to the invention contains a heavy chain containing the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO: 3; HCDR1 AB79), ISWNGGKT (SEQ ID NO: 4; HCDR2 AB79), ARGSLFHDSSGFYFGH (SEQ ID NO: 5 ; HCDR3 AB79) or variants of these sequences containing up to three amino acid substitutions and a light chain containing the following CDR amino acid sequences: SSNIGDNY (SEQ ID NO: 6; LCDR1 AB79), RDS (SEQ ID NO: 7; LCDR2 AB79) and QSYDSSLSGS ( SEQ ID NO: 8; LCDR3 AB79) or variants of these sequences containing up to three amino acid substitutions. In some embodiments, the anti-CD38 antibody comprises a heavy chain comprising the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO: 3; HCDR1 AB79), ISWNGGKT (SEQ ID NO: 4; HCDR2 AB79), and ARGSLFHDSSGFYFGH (SEQ ID NO: 5; HCDR3 AB79). In some embodiments, the antibody comprises a light chain containing the following CDR amino acid sequences: SSNIGDNY (SEQ ID NO:6; LCDR1 AB79), RDS (SEQ ID NO:7; LCDR2 AB79) and QSYDSSLSGS (SEQ ID NO:8; LCDR3 AB79) . In some embodiments, the antibody comprises a heavy chain containing the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO: 3; HCDR1 AB79), ISWNGGKT (SEQ ID NO: 4; HCDR2 AB79), ARGSLFHDSSGFYFGH (SEQ ID NO: 5; HCDR3 AB79) , and a light chain containing the following CDR amino acid sequences: SSNIGDNY (SEQ ID NO: 6; LCDR1 AB79), RDS (SEQ ID NO: 7; LCDR2 AB79) and QSYDSSLSGS (SEQ ID NO: 8; LCDR3 AB79). In some embodiments, the antibody comprises a heavy chain comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 9. Accordingly, the VH chain may contain CDR sequences as defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the sequence may have at least 80% sequence identity to SEQ ID NO: 9. Accordingly, the VH chain may contain CDR sequences as defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5 and the remainder of the sequence may have at least 85% sequence identity to SEQ ID NO: 9. Accordingly, the VH chain may contain CDR sequences as defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5 and the remainder of the sequence may have at least 90% sequence identity to SEQ ID NO: 9. Accordingly, the VH chain may contain CDR sequences as defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5 and the remainder of the sequence may have at least 95% sequence identity to SEQ ID NO: 9. Accordingly, the VH chain may contain CDR sequences as defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5 and the remainder of the sequence may have at least 97% sequence identity to SEQ ID NO: 9. Accordingly, the VH chain may contain CDR sequences as defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5 and the remainder of the sequence may have at least 99% sequence identity to SEQ ID NO: 9.

[00167] In some embodiments, the antibody comprises a heavy chain comprising the heavy chain variable (VH) region amino acid sequence of SEQ ID NO: 9.

EVQLLESGGGLVQPGGSLRLSCAASGFTFDDYGMSWVRQAPGKGLEWVSDISWNGGKTHYVDSVKGQFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGSLFHDSSGFYFGHWGQGTLVTVSSASTKGPSVFPLA (SEQ ID NO:9).

[00168] In some embodiments, the antibody comprises a light chain comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:10. Accordingly, the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 80% sequence identity to SEQ ID NO: 10. Accordingly, the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 85% sequence identity to SEQ ID NO: 10. Accordingly, the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 90% sequence identity to SEQ ID NO: 10. Accordingly, the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 95% sequence identity to SEQ ID NO: 10. Accordingly, the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 97% sequence identity to SEQ ID NO: 10. Accordingly, the VL chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the VL sequence may have at least 99% sequence identity to SEQ ID NO: 10.

[00169] In some embodiments, the antibody comprises a light chain comprising the light chain variable region (VL) amino acid sequence of SEQ ID NO: 10.

QSVLTQPPSASGTPGQRVTISCSGSSSNIGDNYVSWYQQLPGTAPKLLIYRDSQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQSYDSSLSGSVFGGGTKLTVLGQPKANPTVTLFPPSSEEL (SEQ ID NO:10).

[00170] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of chain VHSEQ ID NO: 9 or a variant thereof, as described herein, and a light chain comprising the amino acid sequence of chain VL SEQ ID NO: 10 or a variant thereof, as described in this document.

[00171] As will be appreciated by those skilled in the art, variable heavy and light chains can be attached to human IgG constant domain sequences, typically IgG1, IgG2 or IgG4.

[00172] In some embodiments, the antibody comprises a heavy chain (HC) comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:11. Accordingly, the heavy chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the heavy chain sequence may have at least 80% sequence identity to SEQ ID NO: 11. Accordingly, the heavy chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the heavy chain sequence may have at least 85% sequence identity to SEQ ID NO: 11. Accordingly, the heavy chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the heavy chain sequence may have at least 90% sequence identity to SEQ ID NO: 11. Accordingly, the heavy chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the heavy chain sequence may have at least 95% sequence identity to SEQ ID NO: 11. Accordingly, the heavy chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the heavy chain sequence may have at least 97% sequence identity to SEQ ID NO: 11. Accordingly, the heavy chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the heavy chain sequence may have at least 99% sequence identity to SEQ ID NO: 11.

[00173] In some embodiments, the antibody comprises the heavy chain (HC) amino acid sequence of SEQ ID NO: 11.

EVQLLESGGGLVQPGGSLRLSCAASGFTFDDYGMSWVRQAPGKGLEWVSDISWNGGKTHYVDSVKGQFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGSLFHDSSGFYFGHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:11).

[00174] In some embodiments, the antibody comprises a light chain (LC) comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:12. Accordingly, the light chain may comprise the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, and the remainder of the light chain sequence may have at least 80% sequence identity to SEQ ID NO: 12. Accordingly, the light chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the light chain sequence may have at least 85% sequence identity to SEQ ID NO: 12. Accordingly, the light chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the light chain sequence may have at least 90% sequence identity to SEQ ID NO: 12. Accordingly, the light chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the light chain sequence may have at least 95% sequence identity to SEQ ID NO: 12. Accordingly, the light chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the light chain sequence may have at least 97% sequence identity to SEQ ID NO: 12. Accordingly, the light chain may contain the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, and the remainder of the light chain sequence may have at least 99% sequence identity to SEQ ID NO: 12.

[00175] In some embodiments, the antibody comprises the light chain (LC) amino acid sequence of SEQ ID NO: 12.

QSVLTQPPSASGTPGQRVTISCSGSSSNIGDNYVSWYQQLPGTAPKLLIYRDSQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQSYDSSLSGSVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS (SEQ ID NO:12).

In some embodiments, the antibody comprises the amino acid sequence HC SEQ ID NO: 11 or a variant thereof, as described herein, and the amino acid sequence LC SEQ ID NO: 12 or a variant thereof, as described herein.

[00176] This invention includes antibodies that bind to both human and macaque CD38 and interact with at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the following amino acid residues: K121, F135, Q139, D141, M142, E239, W241, S274, C275, K276, F284, V288, K289, N290, P291, E292, D293 and S294 of SEQ ID NO: 1 and SEQ ID NO: 2 based on human numbering. Accordingly, the antibody can react with at least 90% of these amino acid residues. Accordingly, the antibody can react with at least 95% of these amino acid residues. Accordingly, the antibody can react with at least 97% of these amino acid residues. Accordingly, the antibody can react with at least 98% of these amino acid residues. Accordingly, the antibody can react with at least 99% of these amino acid residues. Accordingly, the antibody may react with at least 14 (e.g., at least 15 or at least 16) of the following amino acids: K121, F135, Q139, D141, M142, E239, W241, S274, C275, K276, F284, V288 , K289, N290, P291, E292, D293 and S294 from SEQ ID NO: 1 and SEQ ID NO: 2 based on human numbering.

[00177] In some embodiments, the antibodies are full-length. By “full-length antibody” as used herein is meant the structure that constitutes the natural biological form of the antibody, including variable and constant regions, including one or more modifications, as described herein.

[00178] Alternatively, antibodies can have a variety of structures, including, but not limited to, antibody fragments, monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as “antibody mimetics”), chimeric antibodies , humanized. antibodies, fusion antibodies (sometimes called "antibody conjugates") and fragments thereof, respectively. Specific antibody fragments include, but are not limited to, (i) a Fab fragment consisting of VL, VH, CL and CH1 domains, (ii) an Fd fragment consisting of VH and CH1 domains, (iii) an Fv fragment consisting of VL domains and VH of one antibody; (iv) a dAb fragment (Ward et al. (1989) Nature 341: 544-546) which consists of one variable region, (v) isolated CDR regions, (vi) fragments of F(ab')2, a divalent fragment containing two related Fab fragment; (vii) single-chain Fv (scFv) molecules, where the VH domain and the VL domain are linked by a peptide linker that allows the two domains to associate to form an antigen binding site (Bird et al. (1988) Science 242: 423-426, Huston et al. ( 1988) Proc. Natl. Acad. Sci. USA 85: 5879-5883), (viii) bispecific single chain Fv (WO 03/11161) and (ix) “diabodies” or “tribodies”, polyvalent or multispecific fragments constructed by fusion genes (Tomlinson et al. (2000) Methods Enzymol. 326: 461-479; WO94/13804; Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90: 6444-6448).

[00179] Accordingly, the antibody may be a Fab fragment. Accordingly, the antibody may be an Fv fragment. Accordingly, the antibody may be an Fd fragment. Accordingly, the antibody structure may comprise selected CDR regions. Accordingly, the antibody may be an F(ab')2 fragment. Accordingly, the antibody may be a scFv fragment.

[00180] In some embodiments, the antibodies do not cause significant levels of red blood cell depletion and/or platelet depletion after 1 day, 2 days, 4 days, 8 days, 10 days, 15 days, 20 days, 25 days, and/or 30 days after introduction.

[00181] The term “significant level of cell population depletion” may refer to a level of cell population depletion that has adverse consequences for a subject.

[00182] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 1 day after administration.

[00183] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 2 days after administration.

[00184] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 4 days after administration.

[00185] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 8 days after administration.

[00186] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 10 days after administration.

[00187] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 15 days after administration.

[00188] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 20 days after administration.

[00189] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 25 days after administration.

[00190] In some embodiments, the antibodies do not cause a significant level of red blood cell depletion and/or platelet depletion 30 days after administration.

[00191] Accordingly, antibodies for use according to this invention may cause less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less 1% depletion of red blood cell population after treatment. Accordingly, antibodies for use according to this invention may cause less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% depletion platelet populations after treatment.

Antibody modifications

[00192] This invention also provides variants of anti-CD38 antibodies. That is, there are a number of modifications that can be made to the antibodies of the invention, including, but not limited to, CDR (affinity maturation) amino acid modifications, Fc region amino acid modifications, glycosylation variants, other types of covalent modifications, etc.

[00193] The term “variant” means a polypeptide that is different from the original polypeptide. Amino acid variants may contain amino acid substitutions, insertions, and deletions. In general, variants can include any number of modifications as long as the function of the protein is still present, as described herein. That is, for example, in the case of amino acid variants generated by the AB79 CDR, the antibody must still specifically bind to both human and macaque CD38. The term “variant Fc region” means an Fc sequence that differs from the wild-type Fc sequence or the original Fc sequence based on at least one amino acid modification. An Fc variant may refer to the Fc polypeptide itself, compositions containing a variant Fc polypeptide, or an amino acid sequence. If amino acid variants are generated with an Fc region, for example, the variant antibodies must support the functions required for the particular use or indication of the antibody. For example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions may be used, such as 1-10, 1-5, 1-4, 1-3, and 1-2 substitutions. Suitable modifications may be made in one or more positions, as generally outlined in, for example, US Patent Application Nos. 11/841,654; 12/341769; US Patent Publication No. 2004013210; 20050054832; 20060024298; 20060121032; 20060235208; 20070148170; and US patents No. 6737056; 7670600; and 6,086,875, all of which are incorporated by reference in their entirety, particularly for specific amino acid substitutions that increase binding to Fc receptors.

[00194] Accordingly, the variant retains the function of the parent sequence, i.e. option is a functional option. Accordingly, an antibody containing a variant sequence maintains the function of the parent antibody, i.e. an antibody containing the variant sequence is capable of binding human CD38. Accordingly, treatment with a variant may cause less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% depletion of the red blood cell population. Accordingly, treatment with a variant may cause less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1% platelet depletion.

[00195] A variant can be viewed in terms of similarity (ie, amino acid residues having similar chemical properties/functions), preferably the variant is expressed in terms of sequence identity.

[00196] Sequence comparisons can be done by eye or, more commonly, using readily available sequence comparison programs. These publicly available and commercially available computer programs can calculate sequence identity between two or more sequences.

[00197] It may be desirable to have from 1 to 5 modifications in the Fc region of wild-type or engineered proteins, as well as, for example, from 1 to 5 modifications in the Fv region. The polypeptide sequence variant will preferably have at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the parent sequences (eg, variable regions, constant regions and/or heavy and light chain sequences for AB79). Accordingly, the variant may have at least 80% sequence identity with the parent sequence. Accordingly, the variant may have at least 85% sequence identity with the parent sequence. Accordingly, the variant may have at least 90% sequence identity with the parent sequence. Accordingly, the variant may have at least 92% sequence identity with the parent sequence. Accordingly, the variant may have at least 95% sequence identity with the parent sequence. Accordingly, the variant may have at least 97% sequence identity with the parent sequence. Accordingly, the variant may have at least 98% sequence identity with the parent sequence. Accordingly, the variant may have at least 99% sequence identity with the parent sequence.

[00198] In one embodiment, sequence identity is determined across the entire sequence. In one embodiment, sequence identity is determined across the entire candidate sequence compared to the sequence set forth herein.

[00199] The term "amino acid substitution" means the replacement of an amino acid with another amino acid at a specific position in the sequence of the parent polypeptide. For example, the S100A substitution refers to a polypeptide variant in which the serine at position 100 is replaced by alanine. Accordingly, the amino acid substitution may be a conservative amino acid substitution. One or more, for example two or three, conservative amino acid substitutions may be suitable. Amino acids with similar biochemical properties can be defined as amino acids that can be replaced by a conservative substitution.

[00200] Unless otherwise expressly indicated herein by reference to a specific individual amino acid, amino acids may be replaced using conservative substitutions as indicated below. An aliphatic polar uncharged amino acid can be a cysteine, serine, threonine, methionine, asparagine or glutamine residue. The aliphatic polarly charged amino acid may be an aspartic acid, glutamic acid, lysine or arginine residue. An aromatic amino acid can be a histidine, phenylalanine, tryptophan or tyrosine residue. Conservative substitutions can be made, for example, in accordance with Table 2 below. Amino acids in the same block in the second column and preferably in the same row in the third column can replace each other:

[00201] Table 2. Conservative Substitutions

ALIPHATIC Non-polar G A P I L V Polar - uncharged C S T M NQ Polar - charged D E K R AROMATIC H F W Y

[00202] The term "amino acid insertion" means the addition of an amino acid at a specific position in the sequence of the parent polypeptide.

[00203] The term “amino acid deletion” means the removal of an amino acid at a specific position in the sequence of the parent polypeptide.

[00204] The terms “parent antibody” and “precursor antibody” mean an unmodified antibody that is subsequently modified to create a variant. In one embodiment, the parent antibody specified herein is AB79. In one embodiment, the parent antibody herein comprises a VH chain having the amino acid sequence of SEQ ID NO: 9 and a VL chain having the amino acid sequence of SEQ ID NO: 10. In one embodiment of the present invention, the parent antibody contains the heavy chain amino acid sequence of SEQ ID NO: 11 and light chain amino acid sequence SEQ ID NO: 12. The parent antibody may refer to the polypeptide itself, compositions that contain the parent antibody, or the amino acid sequence that encodes it. Accordingly, the term “original Fc polypeptide” means an Fc polypeptide that has been modified to form a variant.

[00205] The terms “wild type,” “WT,” and “native” mean an amino acid sequence or nucleotide sequence that occurs in nature, including allelic variants. Wild type protein, polypeptide, antibody, immunoglobulin, IgG, etc. have an amino acid sequence or nucleotide sequence that has not been intentionally modified.

[00206] In some embodiments, one or more amino acid modifications are made to one or more CDRs of the anti-CD38 antibody. In general, only 1, 2, or 3 amino acids are replaced in any single CDR, and typically no more than 4, 5, 6, 7, 8, 9, or 10 substitutions are made in a set of CDRs. However, it should be understood that any combination of no substitutions, 1, 2 or 3 substitutions in any CDR may be independently and not necessarily combined with any other substitution.

[00207] In some cases, amino acid modifications in CDRs are referred to as "affinity maturation." An “affinity matured” antibody is an antibody that has one or more changes in one or more CDRs that result in increased affinity of the antibody for an antigen compared to a parent antibody that does not have these changes. In some cases, it may be desirable to reduce the affinity of an antibody for its antigen.

[00208] Affinity maturation can be performed to increase the binding affinity of an antibody to an antigen by at least about 10% to 50%, 100%, 150% or more, or 1 to 5 times that of the “parent” antibody. Exemplary affinity matured antibodies will have nanomolar or even picomolar affinity for the target antigen. Affinity matured antibodies are prepared by known methods (eg, Marks et al. (1992) Biotechnol. 10: 779-783; Barbas et al. (1994) Proc. Nat. Acad. Sci. USA 91: 3809-3813; Shier et al. (1995) Gene 169: 147-155 , Yelton et al (1995) J Immunol 155: 1994-2004, Jackson et al (1995) J Immunol 154(7): 3310-9 and Hawkins et al (1995) J Immunol 154(7): 3310-9 al (1992) J Mol Biol 226: 889-896).

[00209] Alternatively, amino acid modifications can be made, for example in one or more CDRs of the antibodies of the invention, that are silent, for example, that do not significantly change the affinity of the antibody for the antigen. This may be done for a number of reasons, including optimization of expression (as may be done for nucleic acids encoding antibodies of the invention).

[00210] Thus, CDR and antibody variants are included in the definition of CDRs and antibodies of the invention; that is, the antibodies of the invention may contain amino acid modifications in one or more of the CDRs set forth in SEQ ID NO: 3-8. In addition, as discussed below, amino acid modifications may also be independently and optionally made to any region outside the CDR, including framework and constant regions.

[00211] In some embodiments, variants of the AB79 antibody are described that are specific for human CD38 (SEQ ID NO: 1) and cynomolgus CD38 (SEQ ID NO: 2). This antibody consists of six CDRs, where each CDR of this antibody may be different from SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 and/or SEQ ID NO 8: 0, 1 or 2 amino acid substitutions.

[00212] In addition to the modifications described above, other modifications may be made. For example, molecules can be stabilized by the inclusion of disulfide bridges linking the VH and VL domains (Reiter et al. (1996) Nature Biotech. 14: 1239-1245). In addition, there are many covalent modifications of antibodies that can be performed, as described below.

[00213] Covalent modifications of antibodies are included within the scope of this invention and are usually, but not always, performed post-translationally. For example, several types of covalent modifications of the antibody construct are introduced into the molecule by reacting specific amino acid residues of the antibody construct with an organic derivatizing agent that is capable of reacting with selected side chains or terminal N- or C- residues.

[00214] In some embodiments, the anti-CD38 antibody of the present invention specifically binds to one or more residues or regions in CD38, but also does not cross-react with other CD38 homologous proteins, such as BST-1 (bone marrow stromal cell antigen-1 ) and/or Mo5, also called CD157.

[00215] Typically, no cross-reactivity means less than about 5% relative competitive inhibition between molecules when assessed by ELISA and/or FACS analysis using sufficient quantities of molecules under suitable assay conditions.

Inhibition of CD38 activity and reduction of side effects

[00216] The disclosed antibodies may find use in blocking a ligand-receptor interaction or inhibiting the interaction of a receptor component. The anti-CD38 antibodies of the invention may be "blocking" or "neutralizing". The term “neutralizing antibody” refers to an antibody whose binding to CD38 results in the inhibition of the biological activities of CD38, such as its ability to interact with ligands, enzymatic activity, signaling ability and, in particular, its ability to cause activation of lymphocytes. Inhibition of CD38 biological activity can be assessed using one or more of several standard in vitro or in vivo assays known in the art.

[00217] The terms “inhibits binding” and “blocks binding” (eg, when referring to inhibiting/blocking the binding of a CD38 antibody to CD38) cover both partial and complete inhibition/blocking. Inhibiting/blocking the binding of a CD38 antibody to CD38 may reduce or alter the normal level or pattern of cellular signaling that occurs when an anti-CD38 antibody binds to CD38 without inhibition or blocking. Inhibition and blocking are also intended to include any measurable reduction in the binding affinity of the anti-CD38 antibody to CD38 when contacted by the anti-CD38 antibody compared to a ligand not contacted by the anti-CD38 antibody, for example blocking the binding of the anti-CD38 antibody to CD38 by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99% or 100%. Accordingly, the blocking of anti-CD38 antibody binding to CD38 may be at least about 70%. Accordingly, the blocking of anti-CD38 antibody binding to CD38 can be at least about 80%. Accordingly, the blocking of anti-CD38 antibody binding to CD38 can be at least about 90%.

[00218] The disclosed anti-CD38 antibodies may also inhibit cell growth. The term “inhibits growth” refers to any measurable reduction in the growth of cells when exposed to an anti-CD38 antibody compared to the growth of the same cells not exposed to an anti-CD38 antibody, e.g., inhibition of cell culture growth by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99% or 100%. Accordingly, the growth inhibition may be at least about 70%. Accordingly, growth inhibition may be at least about 80%. Accordingly, growth inhibition may be at least about 90%.

[00219] In some embodiments, the disclosed anti-CD38 antibodies are capable of depleting a population of activated lymphocytes and plasma cells. The term “depletion” as used herein means a measurable decrease in serum levels of activated lymphocytes and/or plasma cells in a subject compared to untreated subjects. Typically, the population will be depleted by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100%. Accordingly, population depletion may be at least 50%. Accordingly, population depletion may be at least 60%. Accordingly, population depletion may be at least 70%. Accordingly, population depletion may be at least 80%. Accordingly, population depletion may be at least 90%. Accordingly, the depletion of the population can be at least 100% As shown in the examples below, one particular advantage that the antibodies of this invention exhibit is the ability of these cells to recover after dosing; that is, as is known for some treatments (eg, with anti-CD20 antibodies), depletion of cell populations can last for long periods of time, causing adverse events. As shown herein, the effect on activated lymphocytes and/or plasma cells can be restored.

[00220] The anti-CD38 antibodies of this invention reduce adverse events compared to prior art anti-CD38 antibodies. In some embodiments, an antibody for use in accordance with this invention, for example, AB79, does not cause TEAE. In some embodiments, an antibody for use in accordance with this invention, for example, AB79, reduces the incidence of TEAEs in a patient population compared to other anti-CD38 antibodies, such as MOR202. TEAEs are generally classified as grades 1, 2, 3, 4, and 5, with grade 1 being the least severe and grade 5 the most severe. Based on FDA standards and other Common Terminology Criteria for Adverse Events (CTCAE) guidelines for oncology drugs (see, for example, https://evs.nci.nih.gov/ftp1/CTCAE/CTCAE_4.03_2010-06-14_QuickReference_5× 7.pdf; and https://ctep.cancer.gov/protocoldevelopment/electronic_applications/ctc.htm and Nilsson and Koke (2001) Drug Inform. J. 35: 1289-1299) describe how such grades are usually determined. 1st degree - mild: asymptomatic or mild symptoms; clinical or diagnostic observations only; intervention is not indicated. 2nd degree - moderate: minimal, local or non-invasive intervention is indicated; limiting age-appropriate instrumental activities of daily living (“ADLs”). Grade 3 is severe or medically significant, but does not pose an immediate threat to life: hospitalization or prolongation of hospitalization is indicated; shutdown; ADL self-care limitation. Grade 4 - life-threatening consequence: urgent intervention is indicated. Grade 5 - death associated with AEs.

[00221] In some embodiments, an antibody for use in accordance with this invention, for example, AB79, reduces the incidence of TEAE in a patient population compared to other anti-CD38 antibodies, such as MOR202. In some embodiments, an antibody for use in accordance with this invention, for example, AB79, reduces the severity of TEA compared to other anti-CD38 antibodies from grade 5 to grade 4. In some embodiments, an antibody for use in accordance with this invention, for example, AB79, reduces the severity of TEAE compared to other anti-CD38 antibodies from grade 4 to grade 3. In some embodiments, an antibody for use in accordance with this invention, for example, AB79, reduces the severity of TEAE compared to other anti-CD38 antibodies from grade 3 to grade 2. In some embodiments, an antibody for use in the present invention, such as AB79, reduces the severity of TEAE compared to other anti-CD38 antibodies from grade 2 to grade 1.

[00222] In some embodiments, an antibody for use according to the present invention, for example, AB79, reduces the severity of one or more TEAEs selected from the group consisting of anemia (including hemolytic anemia), thrombocytopenia, fatigue, infusion-related reactions (IRRs) , leukopenia, lymphopenia and nausea. In some embodiments, an antibody for use in accordance with the present invention, for example, AB79, reduces the occurrence of one or more TEAEs selected from the group consisting of anemia (including hemolytic anemia), thrombocytopenia, fatigue, infusion-related reactions (IRR), leukopenia, lymphopenia and nausea.

[00223] In some embodiments, the anti-CD38 antibody causes less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% , depletion of the red blood cell population. In some embodiments, the AB79 antibody causes less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% depletion of the red blood cell population . In some embodiments, the AB79 antibody results in a depletion of the red blood cell population of less than 10%.

[00224] In some embodiments, the anti-CD38 antibody causes less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% , platelet population depletion. In some embodiments, the AB79 antibody causes less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% depletion of the platelet population . In some embodiments, the AB79 antibody results in a platelet population depletion of less than 10%.

[00225] In some embodiments, a diagnostic test is used to determine the presence and/or extent of anemia, including hemolytic anemia. Diagnostic tests for anemia, including hemolytic anemia, including hemoglobin levels. Typically, hemoglobin levels are interpreted as follows: (i) very mild/no anemia: ≥12.0 g/dL, (ii) mild: 10–12 g/dL, (iii) moderate: 8–10 g/dL, (iv) ) severe: 6-8 g/dl, and (v) very severe: ≤ 6 g/dl. Other diagnostic tests for anemia, including hemolytic anemia, include measuring haptoglobin levels. In general, a haptoglobin level ≤ 25 mg/dL indicates the presence of anemia, including hemolytic anemia. Other diagnostic tests include the direct antiglobulin test (DAT) (also called the direct Coombs test), which is used to determine whether red blood cells are coated in vivo with immunoglobulin, complement, or both.

[00226] In some embodiments, a diagnostic test is used to determine the presence and/or extent of thrombocytopenia. Typically, a diagnostic test for thrombocytopenia involves measuring the number of platelets per microliter (µL) of blood. Typically 150 x 10 ³ -450 x 10 ³ platelets per µl of blood are present. Typically, thrombocytopenia is diagnosed when there are <150 × 10 ³ platelets per μl of blood. Mild thrombocytopenia is usually diagnosed when there are 70-150 × 10 ³ platelets per μl of blood. Moderate thrombocytopenia is usually diagnosed when there are 20-70 × 10 ³ platelets per μl. Severe thrombocytopenia is usually diagnosed when there are <20 x 10 ³ platelets per μl of blood.

Indications for the disease

[00227] The antibodies, methods and dosage units of the invention find use in a variety of applications, including the treatment or amelioration of diseases associated with CD38.

[00228] CD38 is expressed in immature hematopoietic cells, decreased in mature cells, and re-expressed at high levels in activated lymphocytes and plasma cells. For example, high CD38 expression is observed in activated B cells, plasma cells, activated CD4+ T cells, activated CD8+ T cells, NK cells, NKT cells, mature dendritic cells (DCs), and activated monocytes. Certain conditions are associated with cells expressing CD38, and certain conditions are associated with overexpression, high expression density, or increased expression of CD38 on the cell surface. Whether a population of cells expresses CD38 or not can be determined by methods known in the art, for example, by flow cytometric determination of the percentage of cells in a given population that are labeled with an antibody that specifically binds CD38, or by immunohistochemical assays, as typically described below for diagnostic applications. . For example, a population of cells in which CD38 expression is found in approximately 10-30% of cells may be considered to be weakly positive for CD38; and a population of cells in which CD38 expression is found in more than 30% of cells can be considered as definitely positive for CD38 (as in Jackson et al. (1988) Clin. Exp. Immunol. 72: 351-356), although others can be used criteria to determine whether a cell population expresses CD38. Cell surface expression density can be determined using methods known in the art, such as, for example, flow cytometric measurement of the average fluorescence intensity of cells that have been fluorescently labeled using antibodies that specifically bind CD38.

[00229] The anti-CD38 therapeutic antibodies of this invention bind to CD38-positive cells, resulting in depletion of these cells through multiple mechanisms of action, including the CDC and ADCC pathways.

[00230] It is known in the art that certain conditions are associated with cells expressing CD38, and certain conditions are associated with overexpression, high expression density, or increased expression of CD38 on the surface of cells. Whether a population of cells expresses CD38 or not can be determined by methods known in the art, for example, by flow cytometric determination of the percentage of cells in a given population that are labeled with an antibody that specifically binds CD38, or by immunohistochemical assays, as typically described below for diagnostic applications. . For example, a population of cells in which CD38 expression is found in approximately 10-30% of cells may be considered to be weakly positive for CD38; and a population of cells in which CD38 expression is found in more than 30% of the cells can be considered as definitely positive for CD38 (Jackson et al. (1988) Clin. Exp. Immunol. 72: 351-356), although other criteria can be used, to determine whether the cell population expresses CD38. The density of expression on cell surfaces can be determined using methods known in the art, such as, for example, flow cytometric measurement of the average fluorescence intensity of cells that have been fluorescently labeled using antibodies that specifically bind CD38.

[00231] In one aspect, the present invention provides methods for treating a condition associated with the proliferation of cells expressing CD38, comprising administering to a patient a pharmaceutically effective amount of a disclosed antibody. In some embodiments, the condition is a cancer, and in certain embodiments, the cancer is a hematologic malignancy. In some embodiments, the condition is multiple myeloma, chronic lymphoblastic leukemia, chronic lymphocytic leukemia, plasma cell leukemia, acute myeloid leukemia, chronic myeloid leukemia, B-cell lymphoma, or Burkitt's lymphoma. In some embodiments, the condition is multiple myeloma.

[00232] In some embodiments, the hematologic malignancy is selected from the group of chronic lymphocytic leukemia, chronic myelogenous leukemia, acute myelogenous leukemia, and acute lymphocytic leukemia. In some embodiments, the hematologic malignancy is chronic lymphocytic leukemia. In some embodiments, the hematologic malignancy is chronic myelogenous leukemia. In some embodiments, the hematologic malignancy is acute myelogenous leukemia. In some embodiments, the hematologic malignancy is acute lymphocytic leukemia.

[00233] In some embodiments, the condition is multiple myeloma.

[00234] CLL is the most common leukemia among adults in the Western world. CLL involves clonal expansion of mature lymphocytes involving lymph nodes and other lymphoid tissues with progressive infiltration of the bone marrow and presence in the peripheral blood. The B-cell form (B-CLL) represents the majority of cases.

B-cell form of chronic lymphocytic leukemia (B-CLL)

[00235] B-CLL is an incurable disease characterized by a progressive increase in the number of anergic monoclonal B cell lines that accumulate in the bone marrow and peripheral blood over a long period of time over many years. CD38 expression is considered an independent poor prognostic factor for B-CLL (Hamblin et al. (2002) Blood 99: 1023-9).

[00236] B-CLL is characterized by two subtypes: slow and aggressive. These clinical phenotypes correlate with the presence or absence of somatic mutations in the immunoglobulin heavy chain variable region (IgVH) gene. As used herein, indolent B-CLL refers to a disorder in a subject having a mutated IgVH gene and/or having one or more clinical phenotypes associated with indolent B-CLL. As used herein, the phrase aggressive B-CLL refers to a disorder in a subject who has a non-mutant IgVH gene and/or has one or more clinical phenotypes associated with aggressive B-CLL.

[00237] Today's standard therapy for B-CLL is palliative and primarily involves the use of the cytotoxic drugs chlorambucil or fludarabine. For relapses, combination therapy is often started with fludarabine, cyclophosphamide in combination with rituximab (an anti-CD20 monoclonal antibody) or alemtuzumab (an anti-CD52 monoclonal antibody). In one study, thirty-five patients with relapsed or refractory aggressive B-cell NHL received high-dose chemotherapy (HCT) followed by rituximab 375 mg/m ² once weekly in 4 doses starting on day 40 and repeated for an additional four doses. doses starting from the 180th day. Rituximab infusions were well tolerated with only grade 3/4 infusion-related toxicity. An unexpected adverse event noted in this study was delayed neutropenia in more than half of the patients (19/35 patients with 46 episodes of grade 3 or 4 neutropenia; Kosmas et al. (2002) Leukemia 16: 2004-2015, which is available online at https://www.nature.com/articles/2402639 ). In another study, six patients received alemtuzumab intravenously every other day three times a week for 12 weeks. The dose was gradually increased daily (3, 10, and then 30 mg) until the patient was refractory. The main TEAEs were anemia, neutropenia (6/6 patients each) and thrombocytopenia (5/6 patients) (Ishizawa et al. (2017) Jpn. J. Clin. Oncol. 47(1): 54-60). Thus, there is a critical unmet medical need to treat B-CLL by reducing hematologic side effects. In some embodiments, methods of treating B-CLL using the disclosed anti-CD38 antibodies are provided, and as described below, this may be accomplished using combination therapy, optionally and independently including any of the foregoing drugs.

Multiple myeloma (MM)

[00238] Multiple myeloma (MM) is a B cell lineage malignancy characterized by neoplastic proliferation of plasma cells in the bone marrow. Pharmacological findings in healthy volunteers supported further study of MM (Fedyk et al. (2018) Blood 132:3249, incorporated herein by reference in its entirety). Proliferation of myeloma cells causes many effects, including lytic lesions (holes) in the bone, decreased numbers of red blood cells, production of abnormal proteins (with associated damage to the kidneys, nerves, and other organs), decreased immune system function, and increased levels of calcium in the blood (hypercalcemia). Current treatment options include chemotherapy, preferably associated with autologous stem cell transplantation (ASCT) when possible. These treatment regimens demonstrate moderate responses. However, only minor changes in overall survival are observed, and the median survival is approximately 3 years. Thus, there is a significant unmet need for the treatment of multiple myeloma. In some embodiments, methods of treating multiple myeloma using the disclosed antibodies are provided.

Monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM)

[00239] Monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM) are asymptomatic pre-malignant diseases characterized by the proliferation of monoclonal plasma cells in the bone marrow and the absence of end organ damage.

[00240] Smoldering multiple myeloma (SMM) is an asymptomatic proliferative disease of plasma cells with a high risk of developing symptomatic or active multiple myeloma (Kyle et al. (2007) N. Engl. J. Med. 356(25): 2582-2590) . The international consensus criterion defining SMM was adopted in 2003 and requires that the patient have an M-protein level >30 g/L and/or clonal bone marrow plasma cells >10% (Internat. Myeloma Working Group (2003) Br. J Haematol 121: 749-757). Patients should not have organ or related tissue disorders such as bone lesions or symptoms. Recent studies have identified two subgroups of SMM: i) patients with developing disease and ii) patients with non-developing disease (Internat. Myeloma Working Group (2003) Br. J. Haematol. 121: 749-757).

[00241] SMM resembles monoclonal gammopathy of undetermined significance (MGUS) in that there is no target organ damage (Kyle et al. (2007) N. Engl. J. Med. 356(25): 2582-2590). Clinically, however, SMM is much more likely to develop into active multiple myeloma or amyloidosis after 20 years (78% chance for SMM compared to 21% for MGUS) (Kyle et al. (2007) N. Engl. J. Med. 356 (25): 2582-2590).

[00242] International consensus criteria defining MGUS require that the patient have an M protein level <30 g/L, bone marrow plasma cells <10%, and no organ or related tissue involvement, including bone lesions or symptoms (Internat. Myeloma Working Group (2003) Br. J. Haematol. 121: 749-757).

Systemic light chain amyloidosis

[00243] Amyloidosis belongs to a family of protein misfolding diseases in which various types of proteins aggregate into extracellular insoluble fibrils. These are complex, multisystem diseases. A common type of systemic amyloidosis is systemic light chain amyloidosis (AL). (Gertz et al. (2004) Am. Soc. Hematol. 2004: 257-82). Like multiple myeloma, AL amyloidosis is a neoplasm of plasma cells. AL amyloidosis is a rare, progressive, and fatal disease of older adults caused by a small population of clonal plasma cells in the bone marrow that produce an excess of free monoclonal immunoglobulin light chains. Once in the bloodstream, these pathological light chains misfold, aggregate and deposit as fibrillar material in the internal organs. The amyloid fibril deposits are the same free light chain protein that is secreted by the clonal plasma cell. (Cohen and Comenzo (2010) Am. J. Hematol. 2010: 287-94; Merlini and Bellotti (2003) New England J. Med. 349(6): 583-96; Murray et al. (2010) Blood (ASH Annual Meeting Abstracts 116 (21): abstr 1909). The deposition of amyloid fibrils results in end organ damage and ultimately death. Clonal plasma cell inhibition therapies improve AL amyloidosis disease by removing the factory that produces circulating toxic light chains, which can then improve organ function and survival. No treatment for systemic AL amyloidosis has received regulatory approval. The agents used are those used to treat multiple myeloma. Thus, there is a critical unmet medical need for the treatment of patients with AL amyloidosis, and targeting CD38 to plasma cells is a suitable therapeutic strategy.

Other conditions associated with CD38

[00244] The antibodies, methods and dosage units of the invention find use in a variety of applications, including the treatment or amelioration of diseases associated with CD38, such as diseases and conditions associated with inflammation and immune diseases, especially diseases associated with activated lymphocytes. The anti-CD38 antibodies of the present invention bind to CD38-positive cells, resulting in depletion of these cells, such as activated lymphocytes, through multiple mechanisms of action, including the CDC and ADCC pathways.

[00245] Thus, any autoimmune disease that exhibits either increased expression of CD38 or increased numbers of CD38-expressing cells as a component of the disease can be treated using the antibodies of the invention. These include, but are not limited to, allogeneic islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, antineutrophil cytoplasmic autoantibodies (ANCA), autoimmune adrenal diseases, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune urticaria, Behçet's disease, bullous pemphigoid, cardiomyopathy, Castleman syndrome, abdominal spruce dermatitis, chronic fatigue immune dysfunction syndrome, chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold-related aggravation disease glutinins , Crohn's disease, dermatomyositis, discoid lupus, essential mixed cryoglobulinemia, factor VIII deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barré, Goodpasture syndrome, graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic pulmonary fibrosis , idiopathic thrombocytopenia purpura (ITP), IgA neuropathy, IgM polyneuropathy, immune-mediated thrombocytopenia, juvenile arthritis, Kawasaki disease, lichen, lupus erythematosus, Meniere's disease, mixed connective tissue disease, multiple sclerosis, type 1 diabetes mellitus, myasthenia gravis, pemphigus vulgaris, malignant anemia, polyarteritis nodosa, polychrondritis, polyglandular syndromes, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobinulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Reinauld's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjorgen's syndrome, rejection solid organ transplant, stiff man syndrome, systemic lupus erythematosus, systemic light chain amyloidosis, takayasu arteritis, temporal arteritis/giant cell arteritis, thrombotic thrombocytopenia purpura, ulcerative colitis, uveitis, vasculitis such as dermatitis herpetiformis, vasculitis, vitiligo and Wegner's granulomatosis.

[00246] In particular, some embodiments of the present invention use antibodies to diagnose and/or treat a number of diseases, including, without limitation, autoimmune diseases, including, without limitation, systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), inflammatory bowel disease (IBD), ulcerative colitis, systemic light chain amyloidosis and graft-versus-host disease. In one aspect, the disease is systemic lupus erythematosus (SLE). In one aspect, the disease is rheumatoid arthritis (RA). In one aspect, the disease is inflammatory bowel disease (IBD). In one aspect, the disease is ulcerative colitis. In one aspect, the disease is graft-versus-host disease. In one aspect, the disease is systemic light chain amyloidosis.

[00247] Thus, for example, it is possible to treat patients with high plasma cell counts, such as SLE patients who have high plasma cell counts, as well as RA patients who have been shown not to respond to CD20-based therapy .

Antibody compositions for administration in vivo

[00248] Antibody formulations used in accordance with this invention are prepared for storage by mixing the antibody having the desired purity with optional pharmaceutically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition (1980) Osol, A. Ed.) in the form of lyophilized formulations or aqueous solutions.

[00249] The formulations provided herein may also contain more than one active compound as needed for the particular indication being treated, preferably those having additional activities that do not adversely affect each other. For example, antibodies with other specificities may need to be provided. Alternatively or additionally, the composition may contain a cytotoxic agent, cytokine, growth inhibitor and/or small molecule antagonist. Such molecules are present in the appropriate combination in amounts that are effective for the intended use.

Subcutaneous administration

[00250] The anti-CD38 antibodies described herein, such as AB79, can be administered in sufficiently therapeutically effective doses to permit subcutaneous administration. Subcutaneous administration is a minimally invasive route of administration and is considered the most versatile and therefore desirable route of administration that can be used for short-term and long-term therapy. In some embodiments, subcutaneous administration may be accomplished by injection. In some embodiments, the injection site or device may be changed when multiple injections or devices are required.

[00251] Accordingly, it is much easier for the patient to self-administer subdermal formulations, especially since the composition may have to be taken regularly throughout the patient's life (eg, starting in the child's first year of life). In addition, the ease and speed of subcutaneous administration allows the patient to better comply with the treatment regimen and, if necessary, receive medications faster. Thus, the subcutaneous anti-CD38 antibody compositions presented herein provide significant advantages over the prior art and address several unmet needs.

[00252] In some embodiments, the antibodies of this invention are administered to a subject in accordance with known methods by the subcutaneous route. In some embodiments, the antibodies of this invention can be administered by subcutaneous injection. In specific embodiments, the subcutaneous formulation is administered subcutaneously to the same location of the patient (eg, administered to the upper arm, anterior thigh, lower abdomen, or upper back) for repeated or continuous injections. In other embodiments, the subcutaneous formulation is administered subcutaneously to a different or changed location of the patient. Single or multiple injections of the compositions can be used.

[00253] In some embodiments, the subcutaneous dosage unit forms described herein can be used to treat cancer. In some embodiments, the subcutaneous dosage unit forms described herein can be used for a hematologic malignancy. In some embodiments, the subcutaneous dosage unit forms described herein can be used for multiple myeloma.

[00254] In some embodiments, the antibodies of the invention have increased bioavailability compared to prior art antibodies. In some embodiments, the bioavailability of antibodies of this invention is increased by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% or more compared to a prior art antibody. which binds to human red blood cells. In some embodiments, the bioavailability of antibodies of this invention is 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 250%, or 300% or more compared to a prior art antibody that binds to human red blood cells. Accordingly, bioavailability can be increased by 50%. Accordingly, bioavailability can be increased by 60%. Accordingly, bioavailability can be increased by 70%. Accordingly, bioavailability can be increased by 80%. Accordingly, bioavailability can be increased by 90%.

[00255] In some embodiments, increased bioavailability allows for subcutaneous administration.

[00256] In some embodiments, antibodies of the invention deplete populations of NK cells, B cells, and/or T cells. In some embodiments, the antibodies of the present invention allow for increased depletion of a population of NK cells compared to depletion of a population of B cells or T cells. In some embodiments, the antibodies of the present invention allow for increased depletion of the NK cell population compared to B cells, as well as increased depletion of the NK cell population compared to T cells. In some embodiments, the antibodies of the present invention allow for increased depletion of the NK cell population compared to B cells, as well as increased depletion of the B cell population compared to T cells. In some embodiments, the antibodies of the present invention allow for increased depletion of the NK cell population compared to B cells, and increased depletion of the B cell population compared to T cells. Accordingly, the antibodies of the invention may help to increase the depletion of the CD38 ⁺ cell population compared to CD38 ⁻ cells.

[00257] In some embodiments, the bioavailability of the anti-CD38 antibodies described herein after subcutaneous administration is from at least 50% to at least 80% compared to intravenous administration normalized for the same dose. In some embodiments, the bioavailability of the anti-CD38 antibodies described herein after subcutaneous administration is at least 60% to at least 80% compared to intravenous administration normalized for the same dose. In some embodiments, the bioavailability of the anti-CD38 antibodies described herein after subcutaneous administration is at least 50% to 70% compared to intravenous administration normalized for the same dose. In some embodiments, the bioavailability of the anti-CD38 antibodies described herein after subcutaneous administration is at least 55% to 65% compared to intravenous administration normalized for the same dose. In some embodiments, the bioavailability of the anti-CD38 antibodies described herein after subcutaneous administration is at least 55% to 70% compared to intravenous administration normalized for the same dose.

[00258] In some embodiments, the bioavailability of anti-CD38 antibodies described herein after subcutaneous administration is at least 40%, at least 45%, at least 50%, at least 51%, at least 52%, according to at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69% , at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, according at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84% or at least 85% compared to intravenous administration normalized for the same dose. Accordingly, bioavailability can be at least 50% compared to intravenous administration normalized for the same dose. Accordingly, bioavailability can be at least 60% compared to intravenous administration normalized for the same dose. Accordingly, bioavailability can be at least 70% compared to intravenous administration normalized for the same dose. Accordingly, bioavailability can be at least 80% compared to intravenous administration normalized for the same dose. Accordingly, bioavailability can be at least 90% compared to intravenous administration normalized for the same dose.

[00259] In some embodiments, the present invention provides a method in which the bioavailability of antibodies of the invention after subcutaneous administration is 50-80% compared to intravenous administration normalized for the same dose.

[00260] In some embodiments, the present invention provides a method in which the bioavailability of antibodies of the invention after subcutaneous administration is at least 50% compared to intravenous administration normalized for the same dose.

[00261] In some embodiments, the present invention provides a method in which the bioavailability of antibodies of the invention after subcutaneous administration is at least 55% compared to intravenous administration normalized for the same dose.

[00262] In some embodiments, the present invention provides a method in which the bioavailability of antibodies of the invention after subcutaneous administration is at least 60% compared to intravenous administration normalized for the same dose.

[00263] In some embodiments, the present invention provides a method in which the bioavailability of antibodies of the invention after subcutaneous administration is at least 65% compared to intravenous administration normalized for the same dose.

[00264] In some embodiments, the present invention provides a method in which the bioavailability of antibodies of the invention after subcutaneous administration is at least 70% compared to intravenous administration normalized for the same dose.

[00265] In some embodiments, the present invention provides a method in which the bioavailability of antibodies of the invention after subcutaneous administration is at least 75% compared to intravenous administration normalized for the same dose.

[00266] In some embodiments, the present invention provides a method in which the bioavailability of antibodies of the invention after subcutaneous administration is at least 80% compared to intravenous administration normalized for the same dose.

[00267] In some embodiments, the present invention provides a unit dosage form containing an anti-CD38 antibody as described herein, wherein the anti-CD38 antibody results in a population depletion of less than 10% of red blood cells.

[00268] In some embodiments, the present invention provides a unit dosage form containing an anti-CD38 antibody as described herein, wherein the anti-CD38 antibody results in a population depletion of less than 10% of platelets.

[00269] In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously as a single bolus injection. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously on a monthly basis. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously every two weeks. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously once a week. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously twice per week. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously on a daily basis. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously every 12 hours. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously every 8 hours. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously every six hours. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously every four hours. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously every two hours. In some embodiments, the anti-CD38 antibodies described herein are administered subcutaneously every hour.

[00270] In some embodiments, subcutaneous dosage unit forms are administered at a dose of from about 45 mg to about 1800 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of from about 135 mg to about 1800 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of from about 600 mg to about 1800 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of about 1200 mg to about 1800 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of from about 45 mg to about 1200 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of from about 135 mg to about 1200 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of from about 600 mg to about 1200 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of from about 45 mg to about 135 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of from about 45 mg to about 600 mg. In some embodiments, the subcutaneous dosage unit forms contain an amount sufficient to administer a dose of from about 135 mg to about 600 mg. In some embodiments, the dose is expressed in mg per kilogram of body weight. In some embodiments, the dosage is a once daily dosage.

Unit dosage forms

[00271] In some embodiments, the anti-CD38 therapeutic antibodies are formulated as part of a unit dosage form. In some embodiments, the anti-CD38 antibody comprises a heavy chain comprising the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO: 3; HCDR1 AB79), ISWNGGKT (SEQ ID NO: 4; HCDR2 AB79), and ARGSLFHDSSGFYFGH (SEQ ID NO: 5; HCDR3 AB79) or variants of these sequences containing up to three amino acid substitutions. In some embodiments, the antibody comprises a heavy chain containing the following CDR amino acid sequences: SSNIGDNY (SEQ ID NO: 6; LCDR1 AB79), RDS (SEQ ID NO: 7; LCDR2 AB79), and QSYDSSLSGS (SEQ ID NO: 8; LCDR3 AB79) or variants of these sequences containing up to three amino acid substitutions. In some embodiments, the antibody comprises a heavy chain containing the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO: 3; HCDR1 AB79), ISWNGGKT (SEQ ID NO: 4; HCDR2 AB79), ARGSLFHDSSGFYFGH (SEQ ID NO: 5; HCDR3 AB79) or variants of these sequences containing up to three amino acid substitutions and a light chain containing the following CDR amino acid sequences: SSNIGDNY (SEQ ID NO: 6; LCDR1 AB79), RDS (SEQ ID NO: 7; LCDR2 AB79) and QSYDSSLSGS (SEQ ID NO: 8; LCDR3 AB79) or variants of these sequences containing up to three amino acid substitutions. In some embodiments, the antibody comprises a heavy chain containing the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO: 3; HCDR1 AB79), ISWNGGKT (SEQ ID NO: 4; HCDR2 AB79) and ARGSLFHDSSGFYFGH (SEQ ID NO: 5; HCDR3 AB79) . In some embodiments, the antibody comprises a light chain containing the following CDR amino acid sequences: SSNIGDNY (SEQ ID NO:6; LCDR1 AB79), RDS (SEQ ID NO:7; LCDR2 AB79) and QSYDSSLSGS (SEQ ID NO:8; LCDR3 AB79) . In some embodiments, the antibody comprises a heavy chain containing the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO: 3; HCDR1 AB79), ISWNGGKT (SEQ ID NO: 4; HCDR2 AB79), ARGSLFHDSSGFYFGH (SEQ ID NO: 5; HCDR3 AB79) , and a light chain containing the following CDR amino acid sequences: SSNIGDNY (SEQ ID NO: 6; LCDR1 AB79), RDS (SEQ ID NO: 7; LCDR2 AB79) and QSYDSSLSGS (SEQ ID NO: 8; LCDR3 AB79). In some embodiments, the antibody comprises a heavy chain comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:9. Accordingly, the heavy chain may contain the following CDR amino acid sequences: GFTFDDYG (SEQ ID NO:3; HCDR1 AB79), ISWNGGKT (SEQ ID NO:4; HCDR2 AB79) and ARGSLFHDSSGFYFGH (SEQ ID NO:5; HCDR3 AB79), and the residue of the heavy chain may have at least 80% sequence identity to SEQ ID NO: 9. In some embodiments, the antibody comprises a heavy chain comprising the heavy chain variable (VH) region amino acid sequence of SEQ ID NO: 9.

EVQLLESGGGLVQPGGSLRLSCAASGFTFDDYGMSWVRQAPGKGLEWVSDISWNGGKTHYVDSVKGQFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGSLFHDSSGFYFGHWGQGTLVTVSSASTKGPSVFPLA (SEQ ID NO:9).

[00272] In some embodiments, the antibody comprises a light chain comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:10. Accordingly, the light chain may contain the following CDR sequences: SSNIGDNY (SEQ ID NO: 6; LCDR1 AB79), RDS (SEQ ID NO: 7; LCDR2 AB79) and QSYDSSLSGS (SEQ ID NO: 8; LCDR3 AB79), and the remainder of the light chain may have at least 80% sequence identity to SEQ ID NO: 10. In some embodiments, the antibody comprises a light chain comprising the amino acid sequence of the light chain variable region (VL) of SEQ ID NO: 10.

QSVLTQPPSASGTPGQRVTISCSGSSSNIGDNYVSWYQQLPGTAPKLLIYRDSQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQSYDSSLSGSVFGGGTKLTVLGQPKANPTVTLFPPSSEEL (SEQ ID NO:10).

[00273] In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of VH chain SEQ ID NO: 9 or a variant thereof, as described herein, and a light chain comprising the amino acid sequence of VL chain SEQ ID NO: 10 or a variant thereof, as described in this document.

[00274] As will be appreciated by those skilled in the art, variable heavy and light chains can be attached to human IgG constant domain sequences, typically IgG1, IgG2 or IgG4.

In some embodiments, the antibody comprises a heavy chain (HC) having an amino acid sequence with at least 80% sequence identity to SEQ ID NO:11. Accordingly, the heavy chain may contain the CDR sequences defined in SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, and the remainder of the heavy chain sequence may have at least 80% sequence identity to SEQ ID NO: 11. In some embodiments, the antibody comprises the heavy chain (HC) amino acid sequence of SEQ ID NO: 11.

EVQLLESGGGLVQPGGSLRLSCAASGFTFDDYGMSWVRQAPGKGLEWVSDISWNGGKTHYVDSVKGQFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGSLFHDSSGFYFGHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:11).

[00275] In some embodiments, the antibody comprises a light chain (LC) having an amino acid sequence with at least 80% sequence identity to SEQ ID NO:12. Accordingly, the light chain may comprise the CDR sequences defined in SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, and the remainder of the light chain sequence may have at least 80% sequence identity to SEQ ID NO: 12. In some embodiments, the antibody comprises the light chain (LC) amino acid sequence of SEQ ID NO: 12.

QSVLTQPPSASGTPGQRVTISCSGSSSNIGDNYVSWYQQLPGTAPKLLIYRDSQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCQSYDSSLSGSVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTH EGSTVEKTVAPTECS (SEQ ID NO:12).

[00276] In some embodiments, the antibody comprises the amino acid sequence HC SEQ ID NO: 11 or a variant thereof, as described herein, and the amino acid sequence LC SEQ ID NO: 12 or a variant thereof, as described herein.

[00277] In some embodiments, the anti-CD38 antibody composition is in a unit dosage form. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 45 mg to about 1800 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 135 mg to about 1800 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 600 mg to about 1800 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of about 1200 mg to about 1800 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 45 mg to about 1200 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 135 mg to about 1,200 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 600 mg to about 1200 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 45 mg to about 135 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 45 mg to about 600 mg. In some embodiments, the unit dosage form contains an amount sufficient to administer a dose of from about 135 mg to about 600 mg. In some embodiments, the dose is expressed in mg per kilogram of body weight. In some embodiments, the dosage is a once daily dosage.

[00278] In some embodiments, the anti-CD38 antibody unit dosage forms provided herein may further contain one or more pharmaceutically acceptable excipients, carriers and/or diluents. In some embodiments, the anti-CD38 antibody is provided in the form of a pharmaceutical composition that contains a unit dosage form according to this invention. Accordingly, the pharmaceutical composition may further contain one or more pharmaceutically acceptable excipients, carriers and/or diluents.

[00279] Dosing regimens are adjusted to provide the optimal desired response (eg, therapeutic response). For example, a single dose may be administered, multiple divided doses may be administered over time, or the dose may be proportionally decreased or increased as indicated depending on the therapeutic situation. The compositions may be formulated in unit dosage form for ease of administration and uniformity of dosage. As used herein, unit dosage form refers to physically discrete units suitable as unitary dosages for the subject to be treated; each unit contains a predetermined amount of active compound calculated to provide the desired therapeutic effect, in combination with the required pharmaceutical carrier.

[00280] The specification of the unit dosage forms of this invention is dictated by and directly depends on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the compounding field, such as the active compound for treating the individual.

[00281] Effective doses and dosage regimens for the anti-CD38 antibodies used in this invention depend on the type and severity of the disease or condition being treated and can be determined by those skilled in the art.

[00282] In one embodiment, the anti-CD38 antibody is administered subcutaneously once a week at a dose of about 45 mg to about 1800 mg. Accordingly, the dose for once weekly administration may range from about 135 mg to about 1800 mg. Accordingly, the dose for once weekly administration may range from about 600 mg to about 1800 mg. Accordingly, the dose for once-weekly administration may range from about 1,200 mg to about 1,800 mg. Accordingly, the dose for once weekly administration may range from about 45 mg to about 1200 mg. Accordingly, the dose for once weekly administration may range from about 135 mg to about 1200 mg. Accordingly, the dose for once weekly administration may range from about 600 to about 1200 mg. Accordingly, the dose for once weekly administration may range from about 45 mg to about 135 mg. Accordingly, the dose for once weekly administration may range from about 45 mg to about 600 mg. Accordingly, the dose for once weekly administration may range from about 135 mg to about 600 mg.

[00283] Such administration can be repeated, for example, 1 to 14 times, for example, 3 to 5 times. An exemplary non-limiting range of a therapeutically effective amount of anti-CD38 antibody used in the present invention is from about 45 mg to about 1800 mg. Accordingly, the dose may be from about 135 to about 1800 mg. Accordingly, the dose may be from about 600 to about 1800 mg. Accordingly, the dose may be from about 1200 to about 1800 mg. Accordingly, the dose may be from about 45 to about 1200 mg. Accordingly, the dose may be from about 135 to about 1200 mg. Accordingly, the dose may be from about 600 to about 1200 mg. Accordingly, the dose may be from about 45 to about 135 mg. Accordingly, the dose may be from about 45 to about 600 mg. Accordingly, the dose may be from about 135 to about 600 mg.

[00284] As non-limiting examples, the treatment of this invention can be provided as a once-per-day dosage of the antibody in an amount of from about 45 to about 1800 mg, for example, 45, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580, 600, 620, 640, 660, 680 , 700, 720, 740, 760, 780, 800, 820, 840, 860, 880, 900, 920, 940, 960, 980, 1000, 1020, 1040, 1060, 1080, 1100, 1120, 1140, 1160, 1180, 1200, 1220, 1240, 1260, 1280, 1300, 1320, 1340, 1360, 1380, 1400, 1420, 1440, 1460, 1480, 1500, 1520, 1540, 1560, 1580, 160 0, 1620, 1640, 1660, 1680, 1700, 1720, 1740, 1760, 1780 or 1800 mg per day for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 days , or alternatively at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 weeks after initiation of treatment or any combination thereof, using a single or divided dose every 24, 18, 12, 8, 6, 4, 2 or 1 hour(s), or any combination thereof. Accordingly, the dose for once daily administration may be about 45 mg. Accordingly, the dose for once daily administration may be about 100 mg. Accordingly, the dose for once daily administration may be about 135 mg. Accordingly, the dose for once daily administration may be about 150 mg. Accordingly, the dose for once daily administration may be about 200 mg. Accordingly, the dose for once daily administration may be about 300 mg. Accordingly, the dose for once daily administration may be about 400 mg. Accordingly, the dose for once daily administration may be about 500 mg. Accordingly, the dose for once daily administration may be about 600 mg. Accordingly, the dose for once daily administration may be about 700 mg. Accordingly, the dose for once daily administration may be about 800 mg. Accordingly, the dose for once daily administration may be about 900 mg. Accordingly, the dose for once daily administration may be about 1000 mg. Accordingly, the dose for once daily administration may be about 1100 mg. Accordingly, the dose for once daily administration may be about 1200 mg. Accordingly, the dose for once daily administration may be about 1300 mg. Accordingly, the dose for once daily administration may be about 1400 mg. Accordingly, the dose for once daily administration may be about 1500 mg. Accordingly, the dose for once daily administration may be about 1600 mg. Accordingly, the dose for once daily administration may be about 1700 mg. Accordingly, the dose for once daily administration may be about 1800 mg.

[00285] In one embodiment, the anti-CD38 antibody is administered at a once-weekly dosage of from about 45 mg to about 1800 mg. Accordingly, the dose for once weekly administration may range from about 135 mg to about 1800 mg. Accordingly, the dose for once weekly administration may range from about 600 mg to about 1800 mg. Accordingly, the dose for once-weekly administration may range from about 1,200 mg to about 1,800 mg. Accordingly, the dose for once weekly administration may range from about 45 mg to about 1200 mg. Accordingly, the dose for once weekly administration may range from about 135 mg to about 1200 mg. Accordingly, the dose for once weekly administration may range from about 600 to about 1200 mg. Accordingly, the dose for once weekly administration may range from about 45 mg to about 135 mg. Accordingly, the dose for once weekly administration may range from about 45 mg to about 600 mg. Accordingly, the dose for once weekly administration may range from about 135 mg to about 600 mg. Such administration can be repeated, for example, 1 to 14 times, for example, 3 to 5 times. Administration may be by continuous infusion over a period of 1 to 24 hours, for example 1 to 12 hours. This regimen can be repeated one or more times as needed, for example after 6 or 12 months. Dosage can be determined or adjusted by measuring the amount of a compound of this invention in the blood upon administration, for example, by collecting a biological sample and using anti-idiotypic antibodies that target the antigen-binding region of the anti-CD38 antibody.

[00286] In one embodiment, the therapeutic antibody is formulated at a concentration of 100 mg/ml. In some embodiments, a volume of 1.75 mL, 2.0 mL, 2.25 mL, or 2.5 mL is administered into the thigh, abdomen, or arm. In some embodiments, a volume of 1.75 mL, 2.0 mL, 2.25 mL, or 2.5 mL is administered into the thigh or abdomen. In some embodiments, a volume of 2.25 ml is administered into the thigh or abdomen. In some embodiments, the dose is administered over a 4-, 6-, 8-, or 10-hour period of time. In some embodiments, the dose is administered over an 8-hour period of time. In some embodiments, 2, 4, 6, or 8 doses are administered. In some embodiments, doses are administered every 2 hours.

[00287] In a further embodiment, the anti-CD38 antibody is administered once a week for 2-12 weeks. Accordingly, the antibody can be administered once a week, for example, for 3-10 weeks. Accordingly, the antibody can be administered once a week, for example, for 4-8 weeks. Accordingly, the antibody can be administered once a week, for example, for 5-7 weeks.

[00288] In one embodiment, the anti-CD38 antibody is administered subcutaneously at a frequency that varies over time. Accordingly, the antibody can be administered once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks for a 28-day treatment cycle until unacceptable toxicity is observed or until the subject withdraws from treatment due to other reasons.

[00289] In one embodiment, the anti-CD38 antibody is administered via maintenance therapy, for example, once a week for 6 months or more.

[00290] In one embodiment, the anti-CD38 antibody is administered in a regimen comprising one infusion of the anti-CD38 antibody followed by an infusion of the radioisotope-conjugated anti-CD38 antibody. The scheme can be repeated, for example, after 7-9 days.

[00291] In one embodiment of the present invention, a unit dosage form is provided containing an anti-CD38 antibody as described herein, wherein the anti-CD38 antibody results in less than 10% depletion of the red blood cell population.

[00292] In one embodiment of the present invention, a unit dosage form is provided containing an anti-CD38 antibody as described herein, wherein the anti-CD38 antibody results in less than 10% depletion of the platelet population.

[00293] In some embodiments, an anti-CD38 antibody for use according to the invention is used in combination with one or more additional therapeutic agents, such as a chemotherapeutic agent. Non-limiting examples of DNA-damaging chemotherapeutic agents include topoisomerase I inhibitors (eg, irinotecan, topotecan, camptothecin and their analogs or metabolites, and doxorubicin); topoisomerase II inhibitors (eg etoposide, teniposide and daunorubicin); alkylating agents (eg, melphalan, chlorambucil, busulfan, thiotepa, ifosfamide, carmustine, lomustine, semustine, streptozocin, decarbazine, methotrexate, mitomycin C and cyclophosphamide); DNA intercalators (eg, cisplatin, oxaliplatin and carboplatin); DNA intercalators and free radical generators such as bleomycin; and nucleoside mimetics (eg, 5-fluorouracil, capecitibine, gemcitabine, fludarabine, cytarabine, mercaptopurine, thioguanine, pentostatin, and hydroxyurea).

[00294] Chemotherapeutic agents that interfere with cell replication include: paclitaxel, docetaxel and related analogs; vincristine, vinblastine and related analogues; thalidomide, lenalidomide and related analogues (eg, CC-5013 and CC-4047); protein tyrosine kinase inhibitors (eg, imatinib mesylate and gefitinib); proteasome inhibitors (eg bortezomib); NF-κB inhibitors, including IκB kinase inhibitors; antibodies that bind to proteins overexpressed, misexpressed, or activated in cancer and thereby inhibit cell replication (eg, trastuzumab, rituximab, cetuximab, and bevacizumab); and other inhibitors of proteins or enzymes known to be upregulated, overexpressed, misexpressed, or upregulated in cancer, the inhibition of which inhibits cell replication.

[00295] In some embodiments, antibodies of the invention can be used before, concomitantly, or after treatment with Velcade® (bortezomib).

Treatment options

[00296] In the methods of the invention, therapy is used to provide a positive therapeutic response to a disease or condition. The term “positive therapeutic response” refers to improvement of a disease or condition and/or improvement of symptoms associated with the disease or condition. For example, a positive therapeutic response would refer to one or more of the following improvements in disease: (1) reduction in the number of neoplastic cells; (2) increased neoplastic cell death; (3) inhibition of tumor cell survival; (5) inhibiting (ie, slowing to some extent, preferably stopping) tumor growth; (6) increased patient survival; and (7) some relief of one or more symptoms associated with the disease or condition.

[00297] Positive therapeutic responses for any given disease or condition can be determined using standard response criteria specific to that disease or condition. Tumor response can be assessed for changes in tumor morphology (i.e., total tumor burden, tumor size, etc.) using screening modalities such as magnetic resonance imaging (MRI), radiographic imaging, computed tomography (CT), Skeletal bone scans, endoscopy and tumor biopsy sampling, including bone marrow aspiration (BMA) and counting of tumor cells in the bloodstream.

[00298] In addition to these positive therapeutic responses, a subject undergoing therapy may experience a positive effect in the form of improvement in symptoms associated with the disease. For B-cell tumors, the subject may experience a decrease in so-called B-symptoms, such as night sweats, fever, weight loss and/or urticaria. For precancerous conditions, therapeutic antibody therapy to CD38 may block and/or prolong the time until the development of a related malignancy, such as the development of multiple myeloma in subjects suffering from monoclonal gammopathy of undetermined significance (MGUS).

[00299] Relief of disease can be characterized as a complete response. The term "complete response" refers to the absence of clinically detectable disease with normalization of any previously abnormal radiographic studies, bone marrow and cerebrospinal fluid (CSF), or abnormal monoclonal protein in the case of myeloma.

[00300] Such a response may last 4 to 8 weeks or 6 to 8 weeks after treatment in accordance with the methods of the invention. Alternatively, disease relief may be categorized as a partial response. The term "partial response" may refer to at least about a 50% reduction in the total measured tumor burden (i.e., the number of malignant cells present in the subject, or the measured volume of tumor masses, or the amount of abnormal monoclonal protein) in the absence of new lesions, which can last from 4 to 8 weeks or from 6 to 8 weeks.

[00301] Treatment according to this invention includes a "therapeutically effective amount" of drugs used.

[00302] The terms “therapeutically effective amount” and “therapeutically effective dose” refer to an amount of therapy that is sufficient to reduce or alleviate the severity and/or duration of a disorder or one or more symptoms thereof; preventing the development of the disorder; causing regression of the disorder; preventing relapse, development, occurrence or progression of one or more symptoms associated with the disorder; or enhancing or improving the prophylactic or therapeutic effect(s) of another therapy (e.g., a prophylactic or therapeutic agent) in doses and for periods of time necessary to achieve the desired therapeutic result. The therapeutically effective amount may vary depending on factors such as the disease state, age, sex and weight of the individual, as well as the ability of the drugs to produce the desired response in the individual. A therapeutically effective amount is also an amount such that any toxic or harmful effects of the antibody or antibody portion are outweighed by the therapeutically beneficial effects. A "therapeutically effective amount" of an antibody for tumor therapy can be measured by its ability to stabilize disease progression. The ability of a compound to inhibit cancer can be assessed in an animal model system predictive of efficacy against human tumors.

[00303] Alternatively, this property of the composition can be assessed by testing the compound's ability to inhibit cell growth or induce apoptosis using in vitro assays known to the skilled practitioner. A therapeutically effective amount of a therapeutic compound may reduce tumor size or otherwise alleviate symptoms in a subject. One skilled in the art will be able to determine such amounts based on factors such as the size of the subject, the severity of the subject's symptoms and the specific conditions or route of administration chosen.

Anti-CD38 Antibody Kits

[00304] In another aspect of the invention, kits are provided for treating a disease or condition associated with hematologic malignancies. In one embodiment, the kit contains a dose of an anti-CD38 antibody described herein, such as AB79. In some embodiments, kits provided herein may contain one or more doses of a liquid or lyophilized formulation as provided herein. When kits include a lyophilized formulation of an anti-CD38 antibody described herein, such as AB79, typically the kits will also contain a suitable liquid for reconstitution, such as sterile water or a pharmaceutically acceptable buffer. In some embodiments, the kits may contain an anti-CD38 antibody formulation described herein pre-packaged in a syringe for subcutaneous administration by a healthcare professional or for home use.

[00305] In some embodiments, the kit is for a single administration or dose of an anti-CD38 antibody described herein, such as AB79. In other embodiments, the kit may contain multiple doses of an anti-CD38 antibody described herein, such as AB79, for subcutaneous administration. In one embodiment, the kit may contain an anti-CD38 antibody formulation described herein pre-packaged in a syringe for subcutaneous administration by a healthcare professional or for home use.

Industrial products

[00306] In other embodiments, an article of manufacture is provided containing materials useful for treating the disorders described above. The product consists of a container and a label. Suitable containers include, for example, bottles, vials, syringes and tubes. Containers can be made from various materials such as glass or plastic. The container contains a composition that is effective for treating the condition and may have a port for sterile access (for example, the container may be a bag of intravenous solution or a vial having a stopper that can be pierced with a hypodermic needle). The active agent in the composition is an antibody. The label on or associated with the container indicates that the composition is used to treat the selected condition. The article may further comprise a second container containing a pharmaceutically acceptable buffer, such as phosphate buffered saline, Ringer's solution, or dextrose solution. It may further contain other materials desirable from a commercial and user perspective, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.

EXAMPLES

Example 1: Model characterization of anti-CD38 antibody in cynomolgus monkey

[00307] The anti-CD38 antibody AB79 binds cynomolgus monkey (macaque) CD38, which distinguishes it from daratumumab (Darzalex ^™ ), a cytolytic anti-CD38 monoclonal antibody recently approved for the treatment of multiple myeloma. This unique feature supported the use of macaques for preclinical studies to characterize the pharmacokinetics (PK), pharmacodynamics (PD), and safety of AB79. To this end, assays were developed to measure drug concentrations, immunogenicity, and quantify T, B, and NK lymphocytes in the blood of cynomolgus monkeys. We assessed these parameters in 8 pharmacological and toxicological preclinical studies. Of the cell populations tested, CD38 is most highly expressed on NK cells; Therefore, we hypothesize that the effect of the drug on NK cells is most similar to the effect on the target cells in question, plasmablasts, plasma cells and other activated lymphocytes.

[00308] Data from 8 studies in healthy monkeys using a dose range of 0.03-100 mg/kg were pooled, and mathematical models were developed that describe the pharmacokinetics and exposure-response relationships for each cell type. Depletion of the NK cell population was identified as the most sensitive pharmacodynamic effect of AB79. This population depletion was described using a turnover model (EC50=34.8 μg/ml kill rate) and complete population depletion was achieved at an intravenous dose of 0.3 mg/kg. Intermediate effects were also observed on T cell counts using the direct response model (EC50=9.43 μg/mL) and on B cells using the 4-compartment model (EC50=19.3 μg/mL by depletion rate). . These analyzes confirmed the observation that each of the measured lymphocyte subpopulations was cleared by AB79 at different rates and required different periods of time to deplete the blood compartment population.

[00309] Mathematical models that describe PK and PD data are useful tools for gaining mechanistic and quantitative understanding of the relationship between drug exposure and effect (Friberg et al. (2002) J. Clin. Oncol. 20: 4713-4721; Mager et al (2003) Drug Metab. Dispos. 31: 510-518; Han and Zhou (2011) Ther. Deliv . 2: 359-368). Typical pharmacokinetic features of IgG antibodies, including distribution and elimination, physiological and genetic similarities between monkey and human, can be used to explain the pharmacology of AB79 (Glassman and Balthasar (2014) Cancer Biol. Med. 11: 20-33; Kamath (2016) Drug Discov. Today Technol. 21-22: 75-83). In addition, these models have been successfully applied to predict FA concentrations and PD effects in healthy individuals (Han and Zhou (2011) Ther. Deliv . 2:359–368).

Materials and methods

[00310] Data from monkey studies are presented in Table 3 in chronological order. Single dose studies 2, 7, and 8 were primarily conducted to evaluate the PK and PD of intravenous (IV) and subcutaneous (SC) administered AB79. Repeated dose studies were conducted to assess safety, PK and PD, including two 4-week studies (studies 1 and 3) and three 13-week studies under GLP conditions (studies 4, 5 and 6). In 13-week Study 5, a dosing error occurred. Animals in the lowest dose group received 0.01 mg/kg instead of the intended 0.1 mg/kg at a time (second dose) and then continued at 0.1 mg/kg. This data was added to the dataset with correct information about the doses actually administered. Study 6 repeated the low dose of 0.1 mg/kg QW in Study 5. All animal studies were performed in accordance with the Guide for the Care and Use of Laboratory Animals adopted and published by the US National Institutes of Health.

Table 3. AB79 monkey studies in chronological order.

Study No. Description of the study Number of animals (females, males) Doses (mg/kg) Number of samples per animal (PK/PD) 1 Day 1 (1 mg/kg) + day 28 (2 mg/kg), IV, FC, PD 6 (0, 6) plc, 1, 2 19/10 2 Single dose, i.v., PK, PD 9 (0, 9) plc, 0.3, 3 14/9 3 4 weeks toxic, once a week, IV, FC, PD 12 (4, 8) plc, 1, 30, 100 15/8 4 13 weeks toxic, once every 2 weeks, IV, FC, FD 40 (20, 20) plc, 3, 30, 80 47/29 5 13 weeks toxic, once a week, IV, FC, FD 52 (26, 26) plc, 0.1, 0.3, 1 31/9 6 13 weeks toxic, once a week, IV, FC, FD 20 (20, 0) plc, 0.1 31/10 7 Single dose, i/v/s/c, FC, PD 12 (12, 0) 0.1, 0.3, 1 16/16 8 Single dose, i/v/s/c, FC, PD 24 (24, 0) 0.03, 0.1, 0.3 19/19

IV: intravenous 30-minute infusion (studies 1-4) or bolus (studies 5-8), SC: subcutaneous injection (arm 4 of study 7 and arm 3 of study 8), PK: frequent sampling of PK, PD : Frequent sampling of whole blood samples for flow cytometry analyzes providing data on the number of T, B and NK cells. plc - placebo, "4 weeks" or "13 weeks" describe the length of the treatment period, tox: toxicology study, every 2 weeks: dosing schedule every two weeks.

Bioanalytics

[00311] PK was analyzed using a validated method developed and conducted by Charles River Laboratories (Reno, NV). Briefly, AB79 concentration was measured in monkey serum using an indirect enzyme-linked immunosorbent assay (ELISA). The 96-well microtiter format was coated with anti-idiotypic AB79 antibody. Blanks, standards and quality control (QC) samples containing AB79 at various concentrations were added to the plate and incubated for 55-65 minutes at room temperature (RT). After washing the microtiter plate, peroxidase-conjugated affinity-pure mouse anti-human IgG (AffiniPure Mouse Anti-Human IgG peroxidase, Fcγ-fragment-specific; Jackson ImmunoResearch) was added and incubated on the plate for an additional 55 to 65 minutes. The plate was washed again, tetramethylbenzidine (TMB) was added to the wells to form the chromophore, and color development was stopped by adding stop solution (2 N sulfuric acid). Optical density at 450 nm was measured using a SPECTRAmax® 190 microplate reader (Molecular Devices), and AB79 concentrations were calculated using a 4-parameter logistic weighted (1/y ² ) standard calibration curve. In study 1 (Table 3), the lower limit of quantitation (LLOQ) of AB79 in serum was 0.061 μg/mL and in all other studies it was 0.05 μg/mL.

Determination of antibodies to AB79 (immunogenicity)

[00312] Anti-drug antibody (ADA) screening of monkey serum was analyzed using a qualitative electrochemiluminescence (ECL) method validated and performed by Charles River Laboratories (Reno, NV). Briefly, undiluted serum samples were incubated with 300 mM acetic acid. Acid-dissociated samples were incubated in a mixture of biotinylated AB79, SULFO-TAG-labeled AB79 (Meso Scale Diagnostics, labeled from Charles River Laboratories), and 1.5 M Trizma base to neutralize the acid and form an immune complex. This complex was then added to a streptavidin-coated MSD plate (Meso Scale Diagnostics) and allowed to bind. After washing, the complex was detected by adding MSD Reading Buffer T (Meso Scale Diagnostics) to the plate and then exciting SULFO-TAG™ through the Ru(bpy)3 electrochemical reaction to generate luminescence (light), which was read using an MSD Sector 6000 (Meso Scale Diagnostics). The amount of luminescence was correlated with the level of simian anti-AB79 antibodies present in the serum of individual samples.

Characteristics of blood cells

[00313] To evaluate and compare the level of AB79 binding between humans and monkeys, blood samples from each were collected in sodium heparin tubes. An aliquot of blood (100 μl) was mixed with an appropriate volume of antibody (Figure 1) and incubated for 15-20 minutes at room temperature in the dark. After incubation, 1 ml of BD FACS lysing agent (1X; BD Biosciences; San Jose, CA) was added to lyse the red cells, and the cells were incubated for 10 minutes at room temperature in the dark, then centrifuged, decanted, and resuspended in 1 ml of solution. bovine serum albumin staining buffer (BD Biosciences). Cells were centrifuged a second time, decanted, and 250 μl of Flow Fix (1% paraformaldehyde in Dulbecco's calcium- and magnesium-free PBS) (Life Technologies, Carlsbad, CA) and fluorescence measured by flow cytometric analysis using a FACSCanto™ II flow cytometer (BD Biosciences). Monkey NK cells (CD3-, CD159a+), B cells (CD3-, CD20+) and T cells (CD3+) and NK cells (CD3-, CD16/CD56+), B cells (CD3-, CD19+) were measured. and human T cells (CD3+). The mean fluorescence intensity for AB79 staining for each cell population was converted to soluble soluble fluorescence equivalent units (MOEF) using a standard curve generated using Rainbow Beads (Spherotech; Lake Forest, IL).

[00314] In the studies shown in Table 3, cells were stained and analyzed using a validated method developed and performed by Charles River Laboratories (Reno, Nevada). Monkey blood samples were collected in sodium heparin tubes before and several times after AB79 treatment, and specific lymphocyte populations were measured by flow cytometric analysis using a FACSCanto™ II flow cytometer (BD Biosciences). Commercial antibodies and anti-CD38 antibody (Ab19; US Patent No. 8362211) were titrated to optimal concentrations for staining. Simian CD38+/-, T cell (CD3+), B cell (CD3-/CD20+) and natural killer (NK) cell (CD3-/CD20-/CD16+) populations were identified and lymphocytes quantified using CD45TruCount™ tubes ( BD Biosciences). Approximately 100 μl aliquots of each blood sample were placed into the appropriate well of a 96-well plate and antibodies were added to the indicated volume, mixed and incubated for a minimum of 30 minutes at room temperature in the dark. After incubation, red blood cells were lysed, samples were mixed and incubated at room temperature for an additional 10 minutes in the dark. The plate was centrifuged and the supernatant was decanted. The cell pellet was then resuspended in 1800 μl of staining buffer, the samples were mixed, centrifuged, and the supernatant was decanted. The cell pellet was resuspended in 500 μl of fetal bovine serum staining buffer and approximately 300 μl of the cell suspension was transferred to a 96-well v-bottom plate for analysis. The percentage of NK cells, as well as the percentage of total T cells and B cells, were applied to cell counts obtained using TruCount™ tubes (BD Biosciences; San Jose, CA) and used to determine absolute cell counts for each cell population. In Studies 1–4, CD38+ NK, B, and T cell subsets were assessed at baseline using labeled anti-CD38 antibodies AB79 or Ab19 (Figure 1). Although TSF-19 binds to a different epitope, the results were very similar and are therefore not presented separately. Processed samples were immediately analyzed.

Development of the FC model

[00315] In developing the PC model, one-, two-, and three-component model structures were investigated. The two-component model was clearly superior to the single-component model, as evidenced by goodness-of-fit (GOF) plots and reduction in objective function value (OFV). Based on visual inspection of the diagnostic areas, the introduction of a third component was not necessary to adequately describe the data. Bioavailability ( F ) was modeled using a logit transformation F=exp (PAR)/(1+exp (PAR)) , where PAR denotes the model parameter to ensure that estimates are constrained between 0 and 1. Nonlinear PK at low concentrations was modeled with using a quasi-steady state (QSS) model approximation of the target drug distribution (TMDD) process (Gibiansky and Gibiansky (2009) Expert Opin. Drug Metab. Toxicol. 5: 803-812).

[00316] A schematic representation of the model is shown in FIG. 3C. To approximate the QSS, we assume that the steady-state concentrations of free drug C, target R, and drug complex RC are established very quickly compared to all other processes. This means that the binding process is balanced by the processes of dissociation and internalization, and that the following equation holds in the appropriate units: K _ON *C*R = (K _OFF +K _INT ) * RC where K _ON denotes the binding rate constant and K _OFF the rate constant dissociation, and K _INT internalization rate constant.

[00317] Between-subject variability (BSV) was examined for all parameters and modeled using exponential models of the following type: PAR _i =TVPAR * e ^ETAPAR _i where PAR _i is a person and TVPAR is a typical parameter estimate, and ETAPAR _i is an individual variance estimate i . The ETAPAR _i values were assumed to follow a normal distribution with mean zero. Residuals were fit using a combined additive and proportional error model (Beal and Sheiner (1992) NONMEM User Guides, in University of California CA).

[00318] The following parameters were examined to identify potential covariate effects on AB79 PK: body weight, gender, dose, route of administration, and study.

Development of the PK-PD model

[00319] For each of the three cell types, the development of the PK-PD model was carried out separately. Note that for model development, measurements close to drug administration (<8 hours post-dose) were not used because they were influenced by a nonspecific drug-independent effect, potentially due to the collection of multiple blood samples in a short period of time ( Fig. 4). The PK model and parameter estimates were corrected. Turnover, transit component, and direct response models of various forms have been tested (Friberg et al. (2002) J. Clin. Oncol. 20: 4713-4721; Mager et al. (2003) Drug Metab. Dispos. 31: 510-518) . In the turnover models, drug effects were introduced on cell clearance rates in the form of an Emax-type model with or without Hill factors. In our notation, the Emax model is a function f of the drug concentration c of the following form: f(c)=EMAX*c ^H /(c ^H +C50 ^H ), EMAX denotes the maximum effect, C50 is the concentration at which half the maximum effect is achieved , and H is the Hill factor. In the transit component model (TCM), the drug effect was introduced and tested at different sites: proliferation rate, circulating cells, and the third transit compartment. Combinations of these effects were also tested and whether the data supported a feedback mechanism from circulation to propagation speed. In addition, Emax-type direct response models with and without Hill factors were tested to describe the drug concentration-effect curve.

[00320] Random effect parameters were introduced to assess between-subject variability in baseline cell count, cell production rate (KIN), model transit time (MTT), C50, and EMAX. Individual average baseline cell levels were presented in the data set (column BL). This was used in the model as a typical value. A random effect parameter was added to allow adjustment of the individual's baseline score based on all of the individual's measurements. FD residuals were described using a proportional error model.

[00321] To validate the model during simulations (PK and PK-PD), OFVs, standard errors, GOF plots, and individual prediction versus data plots were used to evaluate the models and compare them to alternatives.

[00322] The following software packages were used: NONMEM (version 7.2), KIWI (version 1.6), Berkeley Madonna (version 8.3.14), PSN (version 4) and R (version 3.3.0).

Dataset preparation

[00323] Data sets from 8 monkey studies were collected, reorganized into a single format, and combined into three separate NONMEM-readable PK-PD data sets. Each of the three data sets contained individual monkey characteristics (study, ID, group, body weight, sex), dosage information, PK, and NK, B, or T cell data. For control animals, only cell counts and not PK data were added to the data sets, assuming that serum AB79 levels were implicitly missing. Time-resolved information about anti-drug immunogenicity (ADA) status, namely TITER containing the result of the quantitative measurement and the ADAF variable with flag 0/1 (ADAF=1 if ADA affects the concentration of AB79, ADAF=0 if not), was added in separate columns to each observation. ADA titers were measured using different method specifications in different studies and therefore values are not quantitatively directly comparable between studies. To use ADA information in a consistent manner across studies, we applied the following procedure for each animal separately: ADA titers that increased at time points later than 7 days from initially measured levels were considered ADA-positive and noted in the data set ( ADAF=1). If a sample was labeled ADA positive at any time point, all samples collected after that time point were also labeled ADA positive from that animal, regardless of the titer measured. ADA-positive observations were not used to estimate parameters during model development. Note that also partial discharge measurements at sampling points of FA concentrations affected by ADA were labeled ADAF=1.

[00324] For cell count data, individual baseline values for each cell type (NK, B and T cells) were calculated as the average of all available pre-dose measurements for a given animal. In most studies this was a single measurement. Each animal's baseline value was then added as an observation during the first dosing event (TIME=0) and as a constant value in the BL column for each observation of the corresponding animal. From this baseline value, the percentage of baseline for each observed cell number was calculated and added to the data set.

Scaling of monkey FC parameters

[00325] The final PK and PK-PD models were used as a starting point to model the PK and PK-PD profiles for the first human clinical trial. Comparative analysis of therapeutic monoclonal antibody data has shown that PK parameters derived from monkey studies can be scaled to predict human PK profiles with reasonable accuracy (Han and Zhou (2011) Ther. Deliv . 2: 359-368). The publication states that using a fixed exponent of 0.85, the clearance of monoclonal antibodies in humans can be reliably predicted. Therefore, this relationship was used to scale the human clearance parameters (CL, Q), while the volume parameters (VC, VP) were scaled using the direct relationship between body masses (BW):

results

Pharmacokinetics of AB79

[00326] The PK data set was pooled from all 8 healthy monkey studies excluding placebo groups (Table 3). In total, the set contained data on 140 animals, of which 58 were males and 82 females. The body weight of the studied animals ranged from 2.1 to 4.7 kg, and the doses ranged from 0.03 to 100 mg per 1 kg of body weight (mg/kg). In one study group 7 and three study groups 8, doses of 0.03, 0.1, 0.3 and 1 mg/kg were administered subcutaneously (total of 15 animals). The combined data set contained 2199 measured PK values exceeding the LLOQ (Figures 3A, 3B). ADA was assessed in parallel with AB79 concentration. 229 FC observations were found to be influenced by ADA (Figure 5). PK samples were most densely sampled after the first dose, and even in long-term toxicology studies, most animals were sacrificed before 98 days. Only Study 4 included recovery groups, and we were able to collect PK data from only 4 animals, 2 from the 80 mg/kg group and one from each of the 30 mg/kg and 3 mg/kg groups ( Fig. 3B ).

[00327] Initially, PK analyzes were performed for each monkey study using standard non-component analysis (NCA) methods. Based on single dose studies (bolus intravenous injection or 30-minute intravenous infusion), the volume of distribution during the terminal phase (Vz) was calculated to range from 64 to 116 ml/kg, clearance from 6.04 to 14.7 ml/kg/ day, and the final half-life period (T1/2) is from 4.75 to 11.2 days. The area under the concentration-time curve (AUC) and maximum concentration (Cmax) values were found to increase proportionally with dose over a wide range. Only the pharmacokinetic profiles of the lowest dose groups (<1 mg/kg, Figures 3D-3F) suggest a nonlinear increase in clearance at concentrations below 0.5 μg/mL, likely caused by target-directed mechanisms (TMDD) (Kamath (2016) Drug Discov Today Technol. 21-22: 75-83). A linear two-compartment model was constructed from data from all monkey studies except the two lowest dose groups (dose >0.3 mg/kg). When the PK of the lowest dose groups was modeled and superimposed on the measured concentrations, it became apparent that the linear model was a better predictor of concentrations (Figures 3D-3F).

[00328] Available pharmacokinetic data following single dose SC administration indicated that Cmax was 70-80% lower in SC groups at the same dose and that AUCs were comparable. No differences in FC parameters were observed between male and female monkeys. The results of this initial analysis were used as a starting point for developing the model.

Development of the FC model

[00329] Model development began with single intravenous dose data, and then the initial model was gradually expanded to include more complex data. Like other therapeutic antibodies, PK generally follows a linear two-component model (Kamath (2016) Drug Discov. Today Technol. 21-22: 75-83). The nonlinear removal component (TMDD), which describes accelerated clearance at low concentrations, was modeled using the quasi-steady state (QSS) approximation (Gibiansky and Gibiansky (2009) Expert Opin. Drug Metab. Toxicol. 5: 803-812). The assumption that the drug-target association process occurs much faster than the processes of drug dissociation, distribution and elimination, as well as target and drug-target complex elimination, leads to a simplified TMDD model (Fig. 3, Table 4). The amount of data at low concentrations was relatively small, so not all parameters were assessed in a single software evaluation run. Therefore, the TMDD model parameters were first estimated by focusing on data from small single dose studies 7 and 8. The resulting TMDD parameter estimates were then kept fixed during the final estimates for the entire data set (Table 4).

[00330] Table 4. Population PK Modeling Results, Parameter Estimates, and Percentage Standard Errors (%SEM)

Final parameter estimate Interindividual variability/
residual variability Parameter Typical value %SEM Magnitude %SEM F 0.227 121 BUT - K _A (l/day) 0.399 20.5 42.1%CV 53.8 KL (l/day) 0.0187 5.17 42.9%CV 20.1 V _C (l) 0.141 3.23 19.8%CV 22.5 Q (l/day) 0.127 14.7 BUT - V _P (l) 0.127 6.45 39.4%CV 20.8 K _INT (1/day) 0.1 FIXED 49.3%CV 36.7* K _SS (1/day) 5.68 38.7* BUT - K _SYN (units/l/day) ^$ 0.04 FIXED BUT - K _DEG (1/day) 0.00452 30.1* BUT - PATH on V _C -0.697 6.51 BUT - adding an RV 3.17E-04 21.2 0.0178 SD - Properties RV 0.0677 1.58 26.0%KB - Minimum objective function value=5748.412

^$ K _SYN CD38 receptor synthesis rate. Because actual concentration measurements or information on the synthesis or degradation rate of CD38 in vivo were not available, “u” was used as a unit for a certain unknown amount of CD38.

* Estimates and standard errors for TMDD parameters were obtained from a separate run that focused on data from the low-dose groups and were then fixed for the final estimate of other PK parameters. NE: not rated.

[00331] Estimates of the absorption parameters K _A and F were obtained by adding data from the s/c groups. All SC data are from the four lower single dose groups from Studies 7 and 8. These lower doses (≤1 mg/kg) cover a clinically relevant range but may limit the generalizability of parameter estimates to higher doses.

[00332] Inter-subject variability (BSV) for PK parameters was described by exponential models. Rate of absorption (K _A ), clearance (CL ), and peripheral volume of distribution (V _P ) have an estimated BSV of about 40% and a central volume of distribution (V _C ) of about 20% (Table 4). Analysis of covariance revealed an effect of route of administration on V _C . The typical V _C value was 0.141 L for intravenous administration and 0.043 L (approximately 70% less) for subcutaneous administration. No other significant covariate effects were identified. Due to limited data at low concentrations, inter-subject and inter-individual variability in TMDD parameter predictions was assessed only for the degree of K _INT internalization (BSV: 49%). Model evaluation based on residual errors, OFV, standard errors, GOF plots, and individual curves confirmed that the final model adequately described AB79 FC in healthy monkeys (Table 4, Fig. 6).

Pharmacodynamics

[00333] The level of AB79 binding to human and monkey blood NK cells, T cells and B cells was compared using flow cytometric analysis. As shown in FIG. 7, monkey lymphocytes had CD38 expression levels based on AB79 molecules of equivalent fluorescence (MOEF) that were slightly lower compared to their human counterparts, but with a similar ratio between cell types, e.g., CD38 expression on NK cells > B cells cells > T cells. These data support the use of this non-human primate species as a relevant model to help predict the potential of AB79 for PD activity in humans.

[00334] For an in-depth quantitative analysis of the relationship between drug exposure (PD) and the degree and duration of cell population depletion (PD), we compiled data sets from PC concentrations, NK cell counts, B cells, and T cells from all 8 monkey studies , including placebo-treated animals when possible (Table 3). Initial characterization of the data set showed that at baseline, T cells had a median of 3732 cells per μL (interquartile range (IQR): 2.881–5.176) and were the most abundant lymphocyte subtype compared to B cells with 1279 cells per μL. µl (IQR: 860.8-1890) and NK cells with 685 cells per µl (IQR: 482.8-970.1). Baseline CD38 expression in these cell populations was assessed in studies 1–4 (Table 3, n=67). 86.7% (SD 11.3) of NK cells expressed CD38 with less variability. In contrast, 58.7% (SD 27.0) of B cells and 34.5% (SD 24.5) of T cells expressed CD38 with greater variability.

[00335] Data from placebo-treated animals showed that the average number of each cell type varied over time between individual animals more than would be expected from within-individual variability (Figure 8). For example, the mean coefficient of variation in B cell counts of individual placebo curves was 27%, but individual mean B cell counts ranged from 436.6 to 4389. In addition, differences were also observed between mean baseline lymphocyte counts in male and female animals, as well as in animals from different studies, which increased variability (Fig. 9). Based on these results, the cell number after treatment was calculated relative to its individual baseline value as a percentage rather than the absolute number of cells at each time point. For example, a value of 33% means that the number of cells in the sample was 1/3 of the base number of cells. This provided standardized values that could be compared across the entire data set.

[00336] The rapid onset of depletion of AB79 binding cells suggests that the initial concentration in the blood leads to a decrease in the number of lymphocytes (Fig. 10). At AB79 IV doses of 0.3 mg/kg, the mean maximum effect on NK cells was 93.9% (i.e., 6.1% of baseline cell count). At 0.1 mg/kg, the maximum depletion of the population was 71% (29% of baseline remained). At doses >0.3 mg/kg, NK cell populations were almost completely depleted in the blood compartment (nadir (range): 1.06% of baseline (0.17, 6.23); Fig. 10A). After a single dose of 0.3 mg/kg, NK cells required approximately 7 days to recover to an average of 50% of baseline, although the kinetics of recovery varied widely between individuals (Figures 10B, 10C). Consistent with these results, NK function was also tested in a subset of animals in Study 7 (n=3/group; Table 3). This experiment showed a dose-dependent decrease with minimal changes in blood NK activity 48 hours after treatment in animals treated with 0.1 mg/kg AB79 (% lysis at an effector:target ratio of 100:1 ± standard deviation; 44.5% ± 23. 6% versus 41.4% ± 25.8%) and almost complete loss of NK activity in animals treated with 1.0 mg/kg (% lysis at an effector:target ratio of 100:1 ± standard deviation; 37.4% ± 10 .3% versus 6.8% ± 12.5%). NK cell function showed recovery after 57 days, at the next time point measured (% lysis at an effector:target ratio of 100:1 ± standard deviation; 16.0% ± 11.9%).

[00337] B cell and T cell populations were less depleted compared to NK cells, consistent with their lower levels of CD38 expression (Figure 7). For example, when AB79 was administered intravenously at 0.3 mg/kg, B cell populations had an average maximum depletion level of up to 45% of baseline, and T cell populations were depleted to 43% of baseline (Figures 10D, 10G). At this dose level, a 50% reduction in B cell numbers from baseline was not achieved in all animals. Only at the highest doses of ≥30 mg/kg were B cell populations almost completely depleted (Figure 10D). T cell populations were depleted to the same extent as B cell populations, but recovery was faster (Figure 10G-I).

[00338] Two studies 7 and 8 compared intravenous and subcutaneous dosing (Figures 10C, 10F, 10I). There were no obvious differences in cell depletion between administration routes. At lower doses (study 8), sustained (>24 h) cell depletion of 50% below baseline values was observed only in the NK cell population, but not in T and B cells; although all cells showed specific cell population depletion at early time points. The time to onset of depletion of the NK cell population was similar across dose groups, regardless of route of administration, and the duration of depletion was dose-dependent. Cell recovery in all test groups was observed by day 57.

FK-FD models

[00339] Separate PK-PD models have been developed to describe the effects of AB79 on NK, B and T cells. During the PK-PD simulations, the PK parameters were kept fixed according to the estimates of the final PK model, and different PD models were tried (see Materials and Methods for details). The NK cell population in peripheral blood was adequately described by a turnover model, and the killing drug effect was related through PC concentration to an Emax type model with depletion rate. In this model, EMAX represents the maximum rate of additional NK cell depletion and the C50 concentration at which the rate of additional NK cell depletion is half maximum. The structural model of PK-PD for NK cells had the following form:

[00340] In the formula , NK represents the actual number of NK cells, K _IN the production rate, and K _OUT the elimination rate in the absence of the drug. Note that with this basic measurement, BL K _OUT is given by the equation K _OUT =K _IN /BL . c represents the concentration of AB79 in the central compartment. When all parameters were assessed simultaneously, the program did not produce consistent results. Individual KIN, EMAX, and C50 scores were highly correlated. Moreover, due to the limited differentiation between the maximum effects of different doses (see previous section) and the large interindividual variability, accurate estimates of all parameters cannot be expected. In a series of evaluations, one or two of the three parameters KIN, EMAX and C50 were fixed at different values, while the others were evaluated. Stable performance and acceptable goodness of fit were achieved with a fixed KIN of 10,000 and an EMAX of 322. A typical C50 estimate was 29.0 μg/mL (Table 5). In addition, the sensitivity of the selected KIN and EMAX values was tested by selecting different combinations of higher and lower values. Intersubject variability was large: 113% for K _IN NK production rate and 149% for C50, consistent with large individual differences at baseline and between treated animals. The model was evaluated based on residual errors, OFV, standard errors, GOF plots, and individual curves (Table 5, Figure 4).

[00341] Table 5. PD Simulation Results, Parameter Estimates, and Percentage Standard Errors (%SEM)

Final parameter estimate Interindividual variability/
residual variability Parameter Typical value %SEM Magnitude %SEM NK cells KIN (number/day) 10000 FIXED 113%KV 19.2 C50 (µg/ml) 29.0 18.8 149%KV 25.1 EMAX 322 FIXED BUT - Baseline (NK cells)* BUT - 28.3%CV 20.8 Residual NK cells 0.291 2.42 53.9%CV - B cells MTT (days) 8.48 15.4 135%KV 17.4 C50 (µg/ml) 19.5 7.58 BUT - EMAX 2.37 FIXED BUT - Baseline (B cells)* BUT - 24.1%CV 10.7 Residual B cells 0.136 2.20 36.9%CV - T cells C50 (µg/ml) 11.86 7,267 BUT - EMAX 0.4656 6,578 69.46%KB 29.50 Baseline (T cells)* BUT - 29.08%KV 15.50 Residual T cells 0.1343 2,406 36.65%KV -

*For each individual animal, the typical baseline value was calculated as the average of all pre-dose measurements; BUT: not rated

[00342] The transit compartment model was superior to direct response or turnover models for describing AB79-induced depletion of the B cell population. The four transit components were found to be adequate, and the drug effect was described using an Emax-type depletion rate model. Similar to the NK cell killing model, EMAX represents the maximum rate and C50 represents the concentration at which the rate is half the maximum. Thus, the PK-PD structural model for B cells is determined by the following five equations:

[00343] TR _i (i=1-4) represent four transit components. K _TR , K _PROL and K _CIRC are determined by the following equations K _TR =K _PROL =K _CIRC =4/MTT where MTT is the mean transit time (Friberg et al. (2002) J. Clin. Oncol. 20: 4713-4721 ). B represents the number of B cells in the blood and c is the concentration of AB79 in the central component.

[00344] At a fixed EMAX of 2.37, the typical C50 was 19.5 μg/ml and the typical mean transit time (MTT) was 8.48 days (Table 5). The delay of maximum effect relative to the maximum concentration of AB79 was well documented. The model shows that AB79 primarily affects circulating B cells. No additional effects or feedback loops on progenitor cells were required to describe the available data on monkey B cells. Between-subject differences in MTT of 135% and baseline B-cell levels (BASE) of 24.1% indicate large individual differences between animals.

[00345] Drug-induced depletion of the T cell population with rapid recovery has been adequately described using the forward response model: T(c) = BL _T * (1 - EMAX*c/(c+C50)), where T is the actual number of T cells, BL _T - number of T cells at baseline and c concentration of AB79 in the central compartment. The typical C50 was estimated to be 11.86 μg/mL and the typical EMAX was 0.47, indicating that in this case only about half of the T cell population could be depleted by AB79 (Table 5). However, note that the between-subject variability on EMAX was almost 70%. In this model, unlike the NK and B cell depletion models, C50 represents the concentration at which depletion of the T cell population was half-maximal.

[00346] For NK cells, model evaluation of the final PK-PD models for B and T cells based on residual errors, OFV, standard errors, GOF plots, and individual curves confirmed that they adequately described the available monkey data (Table 5, Fig. 4).

PK modeling and depletion of human cell populations

[00347] The monkey PK and PK-PD models were used as a starting point for model-based modeling of human PK and cell count data to support the design and justify selected doses for a first-in-human (FIH) clinical trial in healthy volunteers. To this end, it has been proposed that model structures, including TMDD, derived from monkey data, also describe key features of human FC and subsequent lymphocyte population depletion. To obtain predictions for human PK parameters, we scaled estimates of the following monkey PK parameters: central and peripheral volume of distribution (V _C , V _P ), and clearance (CL) and intercompartmental clearance (Q) with a direct approach for monoclonal antibodies (Han and Zhou (2011) Ther. Deliv . 2: 359-368). AB79 is a fully human monoclonal antibody and therefore we expect less immunogenicity in humans than in monkeys. Therefore, for modeling and simulation, we excluded ADA-positive samples from the data set.

[00348] Using a scaled model, exposure and depletion profiles of NK, B and T cell populations were simulated for single doses via 2-hour infusion (IV) or subcutaneous injection (SC) from 0.0003 to 1.0 mg/kg as planned for the FIH study (Figure 11). Modeling suggests that after an intravenous dose of 0.0003 mg/kg, no observable drug-induced effects on lymphocyte counts or even measurable FA concentrations above the LLOQ would be expected. Because of variability and because of the limited size of the dose groups, it was assumed that the minimal detectable effect of a drug on NK cell counts would be a decrease of at least 10%. At doses of 0.01 mg/kg IV and 0.03 mg/kg SC, NK cell populations were predicted to be depleted to less than 90% of baseline levels.

[00349] At an intravenous dose of 0.3 mg/kg, we predicted depletion of the NK cell population to the remaining 17% of baseline within 3 hours after the end of the infusion and recovery of more than 50% after 11 days (Figure 11). At the same dose, the model predicts that B cell populations are maximally depleted to 67% of baseline after 2.5 days, and T cell populations are immediately depleted to 86% of baseline. For subcutaneous administration of the same dose of 0.3 mg/kg, the model predicted that it would result in less and later maximum killing (nadirs relative to baseline: NK cells 37%, B cells 74%, T cells 94%).

[00350] These preclinical in vitro and in vivo studies demonstrate that the monkey is a suitable animal model for studying the pharmacology of AB79. Densely sampled PK data and NK, B, and T cell counts from eight studies in monkeys with different doses and dosing regimens provide a rich source of data for a comprehensive and quantitative understanding of the relationship between AB79 dose, exposure, and cell population depletion. The generated population PK and PK-PD models adequately describe the observed data and represent a powerful tool for predicting exposure and depletion of lymphocyte populations not only for future studies in monkeys, but also for clinical trials in humans.

[00351] A first-in-human (FIH) single ascending dose trial was conducted in healthy volunteers (www.clinicaltrials.gov: NCT02219256) (Figure 12). The proposed pharmacological effect of AB79 is depletion of the activated lymphocyte population. However, profound and prolonged depletion of the lymphocyte population (enhanced pharmacology) can lead to immune system dysfunction that would not be tolerated in patients or healthy study participants. Therefore, 0.0003 mg/kg was selected as a safe starting IV dose for the FIH study.

[00352] Monkey data showed that depletion of the NK cell population was identified as the most sensitive biological effect. Results from PK-NK modeling helped determine the minimum dose level of 0.01 mg/kg IV at which the most sensitive pharmacological effect (NK cell depletion) could be expected to be detected in humans. New data from the FIH study showed that the overall pattern of dose-dependent and cell type-specific destructive effects of AB79 is consistent with model-based predictions (manuscript in preparation). AB79 was even more effective than expected. For example, at an IV dose of 0.03 mg/kg, the NK cell population in humans was depleted to less than 10% of baseline levels. The median nadir (lowest point of depletion) in monkeys at this dose was 20.0% (Figure 10).

[00353] Three anti-CD38 cytolytic monoclonal antibodies (daratumumab, isatuximab and MOR202) are in clinical development for the treatment of multiple myeloma (van de Donk et al. (2016) Immunol. Rev. 270: 95-112). Daratumumab (Darzalex™, administered as an intravenous infusion) was recently approved for the treatment of multiple myeloma in the United States and non-Hodgkin's lymphoma in Europe. Unlike AB79, daratumumab does not cross-react with monkey CD38. Therefore, comparison of our results with AB79 in cynomolgus monkeys treated with daratumumab was not possible. Moreover, patients with multiple myeloma have high levels of CD38-positive malignant cells, which may require higher effective antibody concentrations for this hallmark of cancer (de Weers et al. (2011) J. Immunol. 186:1840-1848).

[00354] However, it is noteworthy that daratumumab is approved at a weekly intravenous dose of 16 mg/kg for multiple myeloma, even though AB79 achieved complete depletion of peripheral NK cell populations at approximately 1 mg/kg and B cells at 3 mg/kg. kg (Fig. 10).

[00355] Despite the extensive database of 8 monkey studies, a number of limitations were recognized. AB79 effectively depletes NK cell populations even at the lowest dose studied, 0.03 mg/kg. At these low doses, PK quickly falls below the quantitative limit of the bioanalytical assay, preventing exposure-effect relationships from being resolved at lower doses. Moreover, during preclinical development it was recognized that maximum cell population depletion occurs shortly after maximum drug concentration, but the resolution of early phase population depletion is technically limited by the total number of samples and possibly nonspecific cell population depletion due to repeated blood collections (effect blood collection). The blood draw effect was observed as a transient pancytopenia characterized by depletion of cell types that do not bind AB79 and was not dose dependent, suggesting that it was due to blood volume loss resulting from multiple blood draws rather than any specific effects of AB79 (Fig. 13). Consequently, the ability to accurately estimate model parameters, especially for NK cell depletion, was limited, and typical KIN and EMAX values required correction to achieve stable and adequate estimation results.

[00356] The effect of AB79 on tissue plasma cells or plasmablasts cannot be measured. However, like plasma cells and plasmablasts, NK cells have high levels of CD38 on their surface, and the efficiency of cell depletion of a particular lymphocyte subset depends, at least in part, on CD38 expression levels. Therefore, the cytolytic effect of AB79 on plasmablasts and plasma cells may be comparable to the effect on NK cells. Currently, information on the long-term effects of AB79 treatment in monkeys is limited. Only a small subset of animals in the 13-week toxicology studies were studied in the recovery group over a longer period of time, and the majority of animals in all dose groups developed ADAs. Moreover, baseline values and depletion profiles of different lymphocyte subsets varied widely among individuals. Therefore, the long-term effects of AB79 cannot be studied in monkeys and must be studied in humans.

[00357] As new human data become available, it will be interesting to compare human and monkey PK and PD data in detail. Construction of a PK model from human data and comparison with a monkey model will improve the AB79 TMDD model. Data obtained from patient studies will provide insight into how AB79-mediated depletion of B cell lineage populations compares between RA and SLE patients and multiple myeloma patients and healthy subjects. Investigating subject- or disease-related factors that may influence the efficiency of cell population depletion in addition to CD38 expression levels is also important and may lead to personalization of treatment. In addition, careful head-to-head comparison of AB79 with daratumumab and/or other anti-CD38 antibodies in vitro and in vivo will provide valuable information on the pharmacology of anti-CD38 antibodies and their optimal use.

[00358] Extensive pharmacological data and PK and PK-PD models have allowed the characterization of exposure-effect relationships in cynomolgus monkeys. Model-based analysis of NK, B and T cells confirmed and quantified the finding that each of the blood lymphocyte subsets were depleted of antibodies at different rates and required different periods of time to replenish the blood component. These models have proven to be an excellent tool for modeling PK and PD data under different dosing scenarios in clinical trial preparation.

Example 2: CD38+ Cell Depletion with AB79

[00359] CD38 is a cADPR hydrolase expressed on human plasmablasts, plasma cells, NK cells and activated T and B cells, but not on mature platelets or red blood cells based on AB79 binding. In patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), plasma cells and activated B and T cells may play an important role in the development of the disease. Unlike other B cell selective therapies that target CD20 and do not directly deplete plasmablasts that are CD20 ^low/negative , CD38 is expressed at high levels in plasmablasts and plasma cells, making these cells a direct target of AB79. In vitro studies with human blood cells and cell lines have shown that binding of AB79 to CD38 does not result in activation of PBMC cytokines, demonstrating that AB79 is not an agonist, as discussed below. Rather, AB79 mediated the depletion of human B cell lines through ADCC and CDC, and in most cases, cell lines overexpressing CD38 were more susceptible to cell lysis (Figure 14). This is consistent with data obtained in healthy cynomolgus monkeys, where the effectiveness of depletion was correlated with CD38 expression level and AB79 dose level. NK cell populations that express high levels of CD38 were depleted to a greater extent than CD20+ B cell and CD3+ T cell populations that express less CD38 (Figure 15). In vivo , AB79 potently suppressed human B cell responses to antigen in a mouse adoptive transfer model (Figure 16). Together, these data support further investigation of AB79 in autoimmune diseases.

[00360] Human PBMCs were treated with AB79 under several conditions and the release of inflammatory cytokines was measured. Cynomolgus monkeys were used to show the relationship between cell type and dose of AB79, as AB79 cross-reacts with monkey CD38, which is 91% identical to the human protein. A second animal model, adoptively transferred human mouse PBMCs, was used to determine whether AB79 could target human antibody-producing cells.

AB79 binds CD38 and mediates ADCC and CDC

[00361] The number of receptors was determined using FIKIT (DAKO, catalog number K0078) using mouse anti-human CD38 antibodies (clone HIT2) and calculated by converting the mean fluorescence intensity (MFI) of the stained samples to a calibration curve obtained from the MFI 5 population of beads, associated with a certain number of antibody molecules. The absolute number of receptors was calculated by subtracting the MFI of the isotype control (mouse IgG1) from the MFI of the anti-CD38 antibody.

[00362] CCZ was assessed by seeding cell lines at 10,000 cells/well and adding AB79, control IgG, or media. Typically, a 5-point dose-response curve (0.001-10 mg/ml) was constructed. Rabbit complement (2-15 μl; CedarLane Laboratories CL No. 3441) was added to each well except control wells. CytoTox-Glo reagent (Promega, G7571/G7573) was used to determine cytotoxicity by luminescence. Groups tested: cells only; cells + complement; cells + IgG control + complement; cells+AB79+complement. Equation: %RLC:%RLC=100- ((RLU (test) / RLU (complement only)) X 100).

[00363] ADCC was tested by plating 5000 target cells per well (T cell lines) with 50 ml AB79, control IgG, Triton X-100 (1%; Sigma Chemical) or medium alone and 50 ml human effector (E) PBMCs to a T:E cell ratio of 1:25 to 1:50. Typically, a dose-response curve was constructed from 9 points (0.000001-100 nM). Experimental lysis=PBMC+cell line+antibody. Spontaneous lysis=PBMC+cell line without antibodies. Maximum lysis=cell line+Triton X-100. Cytotoxicity was assessed using the CytoTox-Glo™ Cytotoxicity Luminescence Assay (Promega).

AB79 has no agonist activity

[00364] The ability of AB79 treatment to induce cytokine production in human PBMCs was compared with a negative IgG1 isotype control and a positive control, PHA, anti-CD3 (clone OKT3) or anti-CD52 (Campath).

[00365] Soluble AB79 did not increase IL-6 levels (mean ± standard deviation) in PBMCs collected from 4 different subjects after 24 hour incubation compared to the IgG1 isotype control. PHA increased cytokine levels in all subjects, demonstrating that the cells have the ability to produce IL-6. Similar results were obtained when PBMC were stimulated for 48 hours and when tested with IL-2, IL-4, IL-10, GM-CSF, IFNγ and TNFα (data not shown) (Fig. 18).

[00366] The manner in which an antibody is presented to a cell may contribute to the outcome of the antibody:ligand interaction and cellular response (Stebbings et al. (2007) J. Immunol. 179: 3325-3331). Stebbings et al. demonstrated that the maximum cellular response (cytokine release) to an agonistic antibody occurred when the antibody was highly concentrated and attached to the well surface, such as when the antibody was added to the well in solution and the liquid was allowed to evaporate (dry binding) compared to antibodies that were allowed to bind to wells in solution (wet binding) or added directly to PBMC (soluble) (Figure 18A). AB79 did not stimulate cytokine production using either of these approaches (Figure 18B).

[00367] AB79 (100 mg/ml) did not stimulate IL-2, -4, -6, -8, -10, GM-CSF, IFNγ, or TNFα in any of the conditions tested at 24 hours. AB79 did not induce IL-10 or GM-CSF, but both were induced by anti-CD3 (not shown; all values except anti-CD3 were below LLOQ). IL-8 was consistently produced by PBMCs and was not altered by any treatment (data not shown) (Table 6).

[00368] Table 6. AB79 and Cytokine Stimulation

Antibody presentation Absent Isotype AB79 Antibody to CD52 P.H.A. Anti-CD3 antibody IL-2 Soluble: LLOQ LLOQ LLOQ LLOQ 14953 ± 3117 present Wet Bondage LLOQ LLOQ LLOQ LLOQ present 395.0 ± 64.8 Dry binding LLOQ LLOQ LLOQ LLOQ present 167.9 ± 90.1 IL-4 Soluble: 7.3 ± 1.4 4.1 ± 0.8 6.4 ± 3.2 6.9 ± 4.0 50.4 ± 8.9 present Wet Bondage 5.3±08 3.8 ± 1.5 6.4 ± 0.6 8.5 ± 3.7 present 17.8 ± 2.4 Dry binding 4.4 ± 1.8 7.8 ± 2.7 9.1 ± 2.6 10.5 ± 2.1 present 16.2 ± 3.1 IL-6 Soluble: 325.1 ± 65.3 236.0 ± 98.9 170.5 202.0 ± 48.0 18880±0 present Wet Bondage 216.8 ± 95.1 191.2 ± 47.0 194.6 ± 66.0 207.2 ± 60.2 present 902.7 ± 114.1 Dry binding 165.0 ± 79.4 369 ± 143 465.8 ± 230 811.1 ± 473.7 present 500 ± 17 IFNγ Soluble: 1190.2 ± 117.5 857.1 ± 311.7 770.1 ± 203.6 1116.9 ± 330.8 15857 ± 4614.8 present Wet Bondage 1052.8 ± 385.8 657.2 ± 222.6 993.4 ± 198.1 1138.9 ± 339.1 present 5674.6 ± 564.7 Dry binding 768.1 ± 110.0 1583.1 ± 418.6 1927.6 ± 517.6 1827.0 ± 281.5 present 3513.2 ± 708.3 TNFα Soluble: 28.71 ± 8.1 11.6 ± 7.1 59.6 ± 90.1 79.9 ± 18.2 9270.0 ± 0 present Wet Bondage 16.5 ± 3.4 16.1 ± 3.9 14.8 ± 8.8 166.1 ± 21.7 present 2123.3 ± 239.7 Dry binding 16.2 ± 7.4 919.2 ± 77.4 745.1 ± 141 984.4 ± 317.0 present 2231.6 ± 687

A multiplex cytokine assay was used according to the manufacturer's instructions (Bio-Plex Pro ^TM Human Cytokine Standard 8-Plex) to measure concentrations of IL-2, -4, -6, -8, -10, GM-CSF, IFNγ, and TNFα. Abbreviations: lower limit of quantitation LLOQ; n/v, not implemented; PHA, phytohemagglutinin; PBMC, peripheral blood mononuclear cells.

AB79 CD38+ cell depletion

[00369] AB79 binds CD38 with high affinity and mediates CDC and ADCC. AB79 is not an agonist and does not induce cytokine release from human PBMCs. AB79 bound both human and cynomolgus CD38. Lymphocytes from both species had similar cell-specific patterns of CD38 expression with NK cells > B cells > T cells based on mean fluorescence intensity of AB79 staining. Treatment of AB79-depleted monkey lymphocytes in a reversible, cell-specific, and dose-dependent manner. AB79 effectively blocked the human antibody recall response in a mouse adoptive transfer model.

[00370] In cohorts receiving AB79 by subcutaneous injection, dose-dependent reductions in NK cells (Figure 24) and plasmablasts (Figure 25) were observed at doses ≥ 0.1 mg/kg-1 with ≥ 90% reduction in plasmablasts in all subjects receiving a 0.6 mg/kg -1 injection. A 75% reduction in NK cell counts occurred at 0.6 mg kg-1 (data not shown) with a C max of 23.0 ng ml-1 (Table 7). Plasmablast and NK cell levels were reduced from baseline within 8 hours of injection and showed a tmax of 48 hours. The duration of recovery to baseline was variable; recovery to baseline (i.e., within -20% of baseline) for doses of 0.1, 0.3 and 0.6 mg kg-1 required an average of 4, 78 and 50 days, respectively (data not available). presented). There was a minimal decrease in total lymphocytes, B and T cells, cytotoxic T cells, helper T cells, monocytes (Figure 25) and granulocytes, red blood cells and platelets (data not shown).

[00371] Table 7. Summary pharmacokinetic parameters of AB79 after a single subcutaneous injection of AB79 at a dose of 0.6 mg/kg-1 in healthy subjects

Method of administration Dose n t max
(h)
n= 6 C max (ng ml-1) n= 6 AUC _last (ng day-1 ml-1) n= 6 PC 0.6 mg kg ^-1 6 23.87 (7.98, 96.02) ^a 23.0 (67) 90.4 (92)

a n= 5. Values represent average (%CV), except t _max where medians (min., max.) are presented. AUC _last - the area under the serum concentration-time curve from time 0 to the last quantifiable concentration; C max - maximum observed serum concentration; CV - coefficient of variation; IV - intravenously; BUT - not defined; PK - pharmacokinetics; s/c - subcutaneously; t max - time to maximum serum concentration.

AB79 and daratumumab binding data to erythrocytes

[00372] The RBC binding profiles of AB79 and daratumumab were compared. As shown in FIG. 26, there appeared to be a difference in the magnitude of erythrocyte binding (i.e., MFI) between drugs in 3 of 4 donors tested; however, this difference may be attributed to different levels of biotin in each antibody, with daratumumab having 1.6-2.0 times more biotin than AB79. An alternative assay that controls for potential differences in fluorescent antibody labeling is to compare the concentration and binding profile of each antibody, and a useful measure is the concentration at which maximum binding occurs (i.e., maximum specific antigen binding (Bmax)). Bmax is identical for both antibodies in 3 of 4 donors (eg, 1 μg/ml for donor 1). Taken together, these data indicate that both antibodies bind with similar affinities within the current assay resolution limit, which is a multiple of 10. In conclusion, both AB79 and daratumumab bound to red blood cells in this assay with affinities that were within 10 times apart; A 10-fold or greater difference in the binding affinity of these antibodies to red blood cells did not exist within this assay system.

Example 3: Phase 1/2a study to study the safety, tolerability, efficacy, pharmacokinetics and immunogenicity of AB79 administered subcutaneously as a single agent in people with relapsed/refractory (r/r) multiple myeloma (MM)

[00373] The purpose of this study is to evaluate the safety, tolerability, pharmacokinetics (PK), immunogenicity, dose-limiting toxicity (DLT), and maximum tolerated dose (MTD)/recommended dose of a phase 2 study (RP2D) in a phase 1 study and provide a preliminary assessment of the clinical activity of monotherapy AB79 in participants with relapsed and/or refractory multiple myeloma (RRMM). The study included patients with RRMM who had previously received at least a proteasome inhibitor (PI), an immunomodulatory drug (IMid), an alkylating agent, and a steroid. Patients must have refractory disease or be intolerant to at least 1 PI and at least 1 IMiD, and they must have either received 3 or more prior therapies or have received at least 2 prior therapies if one of these therapies involved a combination of a PI and IMiD. For the phase 1b dose escalation portion, prior exposure to an anti-CD38 agent is permitted but not required. In the extension portion of the phase 2a study, patients must also be resistant to at least 1 anti-CD38 monoclonal therapy at any time during treatment. The study was a multicenter study conducted in the United States with approximately 42 participants.

Phase 1

[00374] The Phase 1 study population consists of approximately 24 adult participants aged 18 years or older. Patient characteristics are presented in Table 8.

Table 8. Patient characteristics

Dose Level Characteristics 45 mg
(n=4) 135 mg
(n=3) 300 mg
(n=6) 600 mg
(n=6) Total
(n=19) Median age, years (range) 64.5
(53, 75) 69
(64.74) 63.5 (56.69) 62.5 (60.77) 64
(53.77) Men, % 50 67 67 67 63 ECOG PS 0/1/2, % 85/15/0 25/75/0 33/67/0 0/83/17 26/68/5 ISS I/II/III, lost % 50/25/25/0 33/33/0/33 50/33/17/0 17/50/33/0 37/37/21/5 Median number before therapy direction (range) 4.5 (2-6) 3 (2-6) 3 (2-8) 5.5 (3-8) 4 (2-8) Types of therapy: Based on proteasome inhibitors, n (%) 4 (100) 3 (100) 4 (100) 6 (100) 19 (100) Based on IMID, n (%) 4 (100) 3 (100) 4 (100) 6 (100) 19 (100) Based on monoclonal antibodies, n (%) 1 (25) 0 3 (50) 3 (50) 7 (37) Based on daratumumab, n (%) 1 (25) 0 1 (17) 3 (50) 5 (26) ASCT, n (%) 4 (100) 3 (100) 6 (100) 5 (83) 18 (95) Refractoriness to last therapy, n (%) 3 (75) 1 (33) 4 (67) 4 (67) 12 (63)

[00375] Phase 1 participants are assigned to one of 6 dose-escalating AB79 treatment groups: Cohort 1: 45 mg; cohort 2: 135 mg; cohort 3: 300 mg; cohort 4: 600 mg; cohort 5: 1200 mg; and cohort 6: 1800 mg (Table 9). AB79 is administered subcutaneously once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks for a 28-day treatment cycle until disease progression (DPO), unacceptable toxicity, or withdrawal due to for other factors. Participants may receive premedications 1-3 hours before AB79 administration on each dosing day, such as: dexamethasone (20 mg); acetaminophen (650 to 1000 mg); diphenhydramine (25 to 50 mg) and montelukast (10 mg).

[00376] Table 9: Phase 1 cohorts

Cohort Dosage regimen Cohort 1: 45 mg AB79 Subcutaneous injection of 45 mg AB79 once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks for a subsequent 28-day treatment cycle until the onset of Parkinson's disease, unacceptable toxicity, or other discontinuation reasons. Dose escalation of AB79 to 135 mg can be performed using a 3+3 dose escalation design to determine MTD and/or RP2D. Cohort 2: 135 mg AB79 Subcutaneous injection of 135 mg AB79 once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks for a subsequent 28-day treatment cycle until CR, unacceptable toxicity, or discontinuation for other reasons. . Dose escalation of AB79 to 300 mg can be performed using a 3+3 dose escalation design to determine MTD and/or RP2D. Cohort 3: 300 mg AB79 Subcutaneous injection of 300 mg AB79 once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks for a subsequent 28-day treatment cycle until the onset of Parkinson's disease, unacceptable toxicity, or other discontinuation reasons. Dose escalation of AB79 to 600 mg can be performed using a 3+3 dose escalation design to determine MTD and/or RP2D. Cohort 4: 600 mg AB79 Subcutaneous injection of 600 mg AB79 once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks thereafter for a 28-day treatment cycle until CR, unacceptable toxicity, or discontinuation for other reasons. Dose escalation of AB79 to 1200 mg can be performed using a 3+3 dose escalation design to determine MTD and/or RP2D. Cohort 5: 1200 mg AB79 Subcutaneous injection of 1200 mg AB79 once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks thereafter for a 28-day treatment cycle until CR, unacceptable toxicity, or discontinuation for other reasons. Dose escalation of AB79 to 1800 mg can be performed using a 3+3 dose escalation design to determine MTD and/or RP2D. Cohort 6: 1800 mg AB79 Subcutaneous injection of 1800 mg AB79 once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks thereafter for a 28-day treatment cycle until CR, unacceptable toxicity, or discontinuation for other reasons.

[00377] The total duration of participation in this study is 36 months (3 years). In phase 1, participants who discontinue treatment for any reason other than PD continue to undergo on-site progression-free survival (PFS) surveillance every 4 weeks from the last dose of study drug for 12 months or until DOS, death, loss to follow-up , withdrawal of consent or termination of the study. Participants will be followed 30 days after the last dose of study drug or until the start of subsequent alternative anticancer therapy, whichever occurs first, for follow-up assessment.

Primary efficacy criteria for phase 1

[00378] Primary performance criteria for up to one year include the following:

Number of participants reporting one or more treatment-emergent adverse events (TEEs)

Number of participants with dose-limiting toxicity (DLT): DLT is defined as any of the following events: grade 4 laboratory abnormalities, excluding events that are clearly caused by external causes; non-hematologic TEAEs of grade greater than or equal to (≥) 3, except grade 3 nausea/vomiting, fatigue lasting less than 72 hours, elevated alanine aminotransferase (ALT) or aspartate aminotransferase (AST) levels that resolve to grade less than or equal to (≤) 1 or baseline for 7 days, injection reaction (RI), which responds to symptomatic treatment; hematologic TEAEs, National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE): grade ≥ 4, excluding hemolysis ≥ grade 3, low platelet count grade 3 or higher with clinically significant bleeding; and incomplete recovery from treatment-related toxicity causing a delay of more than (>) 2 weeks in the next scheduled injection before the start of cycle 2 will be considered DLT.

Number of participants with AEE 3 or higher: AE scores are assessed according to NCI CTCAE version 4.03. Grade 1 is assessed as mild; Grade 2 is rated as average; Grade 3 is assessed as severe or medically significant, but not life-threatening; Grade 4 is assessed as life-threatening consequences, and grade 5 is assessed as death associated with AEs.

Number of participants with serious TEAEs

Number of participants with TEAEs leading to treatment discontinuation

Number of participants with TEAEs leading to dose modification

Secondary Performance Criteria

[00379] Secondary performance criteria include the following:

Cmax: maximum observed serum concentration for AB79 [Time frame: cycle 1 and 2: day 1 predose and at multiple time points (up to 168 hours) postdose.]

Tmax: time to maximum observed serum concentration (Cmax) for AB79 [Time frame: cycle 1 and 2: day 1 predose and at multiple time points (up to 168 hours) postdose.]

AUC: area under the serum concentration-time curve from time point 0 to last quantifiable concentration for AB79 [Time Frame: Cycle 1 and 2: day 1 predose and at multiple time points (up to 168 hours) postdose .]

Phase 1: ORR [Time frame: up to 1 year]: ORR is defined as the percentage of participants achieving an ORR of 50% or higher during the study. PR is defined as a ≥50% decrease in serum M-protein and a ≥90% decrease in 24-hour urine M-protein or <200 mg/24 hours.

Percentage of participants with minimal response (MR) [Time frame: up to 1 year]: MR is defined as ≥ 25% but ≤ 49% decrease in serum M-protein and 50-89% decrease in 24-hour urinary M-protein.

Percentage of participants with positive anti-drug antibodies (ADA) [Time frame: up to 1 year].

Results.

[00380] As of March 5, 2019, 19 patients were enrolled in 4 dosing groups (45 mg, 135 mg, 300 mg, and 600 mg) and received at least 1 cycle of AB79. As of March 5, 2019, 9 patients are still under treatment observation, 9 patients have discontinued AB79 due to disease progression, and 1 patient has withdrawn consent.

[00381] As of March 5, 2019, no dose-limiting toxicity (DLT) was reported in evaluable patients in all 4 cohorts (45 mg, 135 mg, 300 mg, and 600 mg). There were no injection site reactions or systemic infusion reactions. There were no reports of drug-related SAEs, study deaths, or AEs that led to study discontinuation. (See Table 10). Additionally, with the exception of 1 transient treatment-related event of grade 3 decrease in neutrophil count and 1 transient anemia, there were no remarkable laboratory findings. Disease progression in one patient was associated with decreased grade 3 platelet levels, anemia, and increased creatinine. This study is ongoing and patients continue to be followed.

[00382] Table 10. NEV data

Accessed January 9, 2019
N (%) 45 mg
(n=4) 135 mg
(n=3) 300 mg
(n=6) 600 mg
(n=6) Total
(n=19) Any adverse events 4 (100) 3 (100) 6 (100) 5 (83) 18 (95) Any drug-related adverse event 3 (75) 2 (67) 3 (50) 3 (50) 11 (58) NEVE grade 3 or more 2 (50) 1 (33) 1 (17) 2 (33) 6 (32) TEAE grade 3 or more associated with the use of the drug 1 (25) 0 1 (17) 0 2 (11) SAE 1 0 0 0 1(5) Drug-related SAE 0 0 0 0 0 Discontinuation due to adverse events 0 0 0 0 0 Dose-limiting toxicity 0 0 0 0 0 Death during research 0 0 0 0 0

[00383] As of March 5, 2019, the most common (>2 patients) TEAEs by preferred condition regardless of causation in all 4 cohorts were anemia (n=7 patients), insomnia (n=5 patients each), upper respiratory tract infections, dizziness, headache and hypertension (n=4 patients each), and diarrhea, fatigue, decreased appetite and muscle cramps (n=3 patients each). All AEs were grade 1 or 2, with the exception of anemia (n=2 patients), diarrhea, decreased platelet count, decreased neutrophil count, increased creatinine, headache, hypertension, and musculoskeletal pain (n=1 patient each), which were grade 3; only 1 case of low neutrophil count and 1 case of anemia were reported to be related to the study drug. There were no cases of RSI, cytokine release syndrome, or injection site reactions. Thus, the present invention provides a safe anti-CD38 antibody for clinical use compared to previous anti-CD38 antibodies such as daratumumab, isatuximab or MOR202.

[00384] In FIG. 20 shows that exposure to Ab79 administered subcutaneously increased with increasing doses over time, consistent with target-mediated drug clearance.

[00385] Subcutaneous administration of Ab79 reduced the levels of blood plasmablasts (Figure 21), plasmablasts in bone marrow aspirates (Figure 22), and plasma cells in bone marrow aspirates (Figure 23) in a dose-dependent manner. CD38 adhered to target cells in peripheral blood at doses ≥ 45 mg per week and in bone marrow ≥ 300 mg. Levels of target cells in the bone marrow and peripheral blood were decreased in a dose-dependent manner at doses ≤ 300 mg.

[00386] In patients with advanced RRMM, AB79 showed early evidence of antitumor activity, as evidenced by a reduction in disease burden of at least 50% in some patients and long-term disease stabilization in others. Although more data are needed to characterize the clinical benefit of this drug, the new data supports the ongoing development of AB79.

[00387] Two anti-CD38 mAbs are currently in clinical development: IV daratumumab is approved for patients with MM (recurrent and newly diagnosed), and IV isatuximab is in the investigational stage. The most common adverse reactions (≥ 20%) with daratumumab alone or in combination with standard antimyeloma regimens are infusion-related reactions (IRRs), neutropenia, thrombocytopenia, fatigue, nausea, diarrhea, constipation, vomiting, muscle cramps, arthralgia, pain in the back, pyrexia, chills, dizziness, insomnia, cough, shortness of breath, peripheral edema, peripheral sensory neuropathy and upper respiratory tract infections. (Darzalex USPI). Daratumumab may cause severe and/or serious infusion reactions, including anaphylactic reactions, which have been reported in approximately half of all patients (Darzalex USPI). You should also be aware that daratumumab interferes with certain important laboratory tests and may complicate blood compatibility testing. (Darzalex USPI). Isatuximab, a humanized monoclonal antibody against CD38, is also being investigated in multiple myeloma. Reported AEs for isatuximab (≥24%) include infusion-related reactions, nausea, fatigue, dyspnea, and cough, which were typically grade ≤2 (Richter et al. (2016) JCO 34 (15): 8005; Dimopoulos et al. (2018) ) Blood 132 (suppl. 1): ASH abstract 155/ oral presentation)). Therefore, based on current evidence, there remains a need for new agents, including a new generation of CD-38 targeted therapies with greater selectivity, hence greater efficacy, resulting in less toxicity and increased patient convenience, to continue to improve clinical outcomes.

Phase 2

[00388] The Phase 2a study population consists of approximately 18 participants. Dosing and premedications for phase 2a are based on a review of available safety, efficacy, PK, and pharmacodynamics data from previous phase 1 cohorts.

[00389] Table 11: Phase 2 cohorts

Cohort Dosage regimen Cohort 1: TBD AB79 Subcutaneous injection of AB79 once weekly for 8 weeks, then once every 2 weeks for 16 weeks, and then once every 4 weeks thereafter for a 28-day treatment cycle until CR, unacceptable toxicity, or discontinuation for other reasons. The dose of AB79 for this phase is determined based on analysis of available safety, efficacy, PK and pharmacodynamic data obtained from the phase 1 study.

Primary efficacy criteria for phase 2a

[00390] Primary performance criteria for up to one year include the following:

Overall response rate (ORR): ORR is defined as the percentage of participants achieving a partial response (PR) of 50 percent (%) tumor reduction or better during the study. PR is defined as a ≥ 50% decrease in serum M protein and a ≥ 90% decrease in 24-hour urine M protein or less (<) 200 milligrams per (mg/) 24 hours.

Secondary efficacy criteria for phase 2a

[00391] Secondary performance criteria for up to one year include the following:

Phase 2a: number of participants with DLT

Phase 2a: number of participants reporting one or more TEAEs

Phase 2a: number of participants with TEAEs leading to dose modification

Phase 2a: number of participants with TEAEs leading to treatment discontinuation

Phase 2a: number of participants with clinically significant laboratory values

Phase 2a: Number of Participants with Clinically Significant Vital Sign Measurements

Phase 2a: Duration of Response (DOR): DOR is the time from the date of first documentation of the response to the date of the first documented RR. A CR is an increase ≥ 25% from the nadir response value in any of the following: serum M protein (the increase should be ≥ 0.5 g/dL; an increase in the serum M component ≥ 1 g/dL is sufficient to define relapse if initial M -component ≥ 5 g/dL), and/or urine M-protein (increase should be ≥ 200 mg/24 hours), and/or only in participants without measurable serum and/or urine M-protein levels, difference between levels free light chain (FLC) involved/uninvolved (increase must be >10 mg/dL), and only in participants without measurable serum levels and/or urinary M-protein levels and without measurable disease by FLC levels, percentage of plasma cells bone marrow (percentage should be ≥ 10%) or definite development of new bone lesions or soft tissue plasmacytomas, or increase in the size of bone lesions or soft tissue plasmacytomas, and development of hypercalcemia that can be attributed solely to a proliferative disorder of plasma cells.

Phase 2a: Progression-free survival (PFS): PFS is the time from the date of the first dose to the earliest date of PFS. PV is an increase ≥ 25% from the nadir response value in any of the following: serum M protein (the increase should be ≥ 0.5 g/dL; an increase in the serum M component ≥ 1 g/dL is sufficient to define relapse if initial M -component ≥ 5 g/dL), and/or urine M-protein (increase should be ≥ 200 mg/24 hours), and/or only in participants without measurable serum and/or urine M-protein levels, difference between levels involved/not involved FLC (increase must be >10 mg/dL), and only in participants without measurable serum levels and/or urinary M-protein levels and without measurable disease by FLC levels, percentage of bone marrow plasma cells (percentage must be ≥ 10%) or definite development of new bone lesions or soft tissue plasmacytomas, or increase in the size of bone lesions or soft tissue plasmacytomas, and development of hypercalcemia that can be attributed solely to a proliferative disorder of plasma cells.

Phase 2a: Overall survival (OS): OS is defined as the time from the date of the first dose to the date of death from any cause.

Phase 2a: Time to Response (TTR): TTR is defined as the time from the date of the first dose to the date of first documentation of response (partial response (PR) or better). PR is defined as a ≥50% decrease in serum M protein and/or a ≥90% decrease in 24-hour urine M protein or <200 mg/24 hours.

Inclusion criteria for phase 1 and phase 2a studies:

[00392] Subjects received the last dose of any of the following treatments/procedures within the specified minimum intervals prior to the first dose of AB79: Myeloma-specific therapy (30 day washout period); antibody therapy (including anti-CD38 antibody) (washout period 120 days); corticosteroid therapy (washout period 30 days); autotransplantation (washout period 90 days); radiation therapy (washout period 30 days); major surgery (washout period 30 days).

[00393] For participants with MM, measurable disease is defined as one of the following: (a) serum M-protein ≥ 500 mg/dL (≥ 5 g/L); (b) M-protein in urine ≥ 200 mg/24 hours; (c) for participants without measurable M-protein by serum protein electrophoresis (SPEP) or urine protein electrophoresis (UPEP), a serum FLC test result with an implicated FLC level of ≥ 10 mg/dL (≥ 100 mg/L), provided that The serum FLC ratio is abnormal.

[00394] Prior therapy must meet all of the following criteria: (a) the participant has previously received at least a proteasome inhibitor (PI), an immunomodulatory drug (IMid), an alkylating agent, and a steroid; (b) the participant is immune or intolerant to at least 1 PI and at least 1 IMid; the patient had either received ≥ 3 previous lines of therapy or had received at least 2 previous lines of therapy if one of these lines included a combination of PI and IMid; (c) the participant may have previously been exposed to an anti-CD38 agent, either as a single agent or in combination, but this is not required.

[00395] In a phase 2a study, participants with MM must also be refractory to at least 1 anti-CD38 monoclonal antibody therapy at any time during treatment. “Refractory” is defined as an increase in M protein or PZ levels of at least 25% during treatment or within 60 days after stopping treatment. “Line of therapy” is defined as 1 or more cycles of a planned treatment program. It may consist of 1 or more planned cycles of single agent or combination therapy, or a sequence of treatments administered in a planned manner. A new line of therapy is initiated when the planned course of therapy is changed to include other therapeutic agents (alone or in combination) as a result of POD, relapse, or toxicity. A new line of therapy is also initiated when the planned observation period of therapy is interrupted by the need for additional treatment of the disease.

Exclusion criteria from the study:

[00396] 1. Sensory or motor neuropathy according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE), grade ≥3.

2. Received an allogeneic stem cell transplant. 3. Received anti-CD38 antibody therapy and did not complete the 120-day washout period before taking AB79. 4. Not cured of adverse reactions to prior myeloma treatment or procedure (chemotherapy, immunotherapy, radiation therapy) to NCI CTCAE grade ≤ 1 or baseline. 5. Clinical signs of damage to the central nervous system (CNS) in MM. 6. Active infection with hepatitis B virus (HBV) or hepatitis C virus (HCV), active infection with HIV or cytomegalovirus (CMV). 7. POEMS syndrome (polyneuropathy, organogalia, endocrinopathy, monoclonal gammopathy and skin changes), monoclonal gammopathy of unknown significance, smoldering myeloma, solitary plasmacytoma, amyloidosis, Waldenström's macroglobulinemia or IgM myeloma. 8. Having a positive Coombs test result during screening.

Inclusion of information in the description of the invention by reference

[00397] The contents of all cited references (including literature references, patents, patent applications and websites) that may be cited in this application are expressly incorporated herein by reference in their entirety for all purposes, as are references cited herein to the same extent as if each individual reference had been specifically and individually indicated to be incorporated by reference in its entirety for all purposes.

Equivalents

[00398] The description may be embodied in other specific forms without departing from the essence or essential characteristics thereof. Thus, the above embodiments are to be considered in all respects as illustrative and not as limiting of the description. Accordingly, the scope of the specification is set forth in the appended claims and not in the preceding specification, and all modifications that fall within the meaning and range of equivalence of the claims are intended to be included within the scope thereof. It is intended that modifications for carrying out the invention that will be obvious to those skilled in the art are within the scope of the appended claims.

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LIST OF SEQUENCES

<110> Takeda Pharmaceutical Company Limited

<120> SUBCUTANEOUS DOSING OF ANTI-CD38 ANTIBODIES

<130> 101588-5010-WO

<150> 62/649489

<151> 2018-03-28

<160> 13

<170> PatentIn version 3.5

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<223> human CD38

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Met Ala Asn Cys Glu Phe Ser Pro Val Ser Gly Asp Lys Pro Cys Cys

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Arg Leu Ser Arg Arg Ala Gln Leu Cys Leu Gly Val Ser Ile Leu Val

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Leu Ile Leu Val Val Val Leu Ala Val Val Val Pro Arg Trp Arg Gln

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Gln Trp Ser Gly Pro Gly Thr Thr Lys Arg Phe Pro Glu Thr Val Leu

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Ala Arg Cys Val Lys Tyr Thr Glu Ile His Pro Glu Met Arg His Val

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Asp Cys Gln Ser Val Trp Asp Ala Phe Lys Gly Ala Phe Ile Ser Lys

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His Pro Cys Asn Ile Thr Glu Glu Asp Tyr Gln Pro Leu Met Lys Leu

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Gly Thr Gln Thr Val Pro Cys Asn Lys Ile Leu Leu Trp Ser Arg Ile

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Lys Asp Leu Ala His Gln Phe Thr Gln Val Gln Arg Asp Met Phe Thr

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Leu Glu Asp Thr Leu Leu Gly Tyr Leu Ala Asp Asp Leu Thr Trp Cys

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Gly Glu Phe Asn Thr Ser Lys Ile Asn Tyr Gln Ser Cys Pro Asp Trp

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Arg Lys Asp Cys Ser Asn Asn Pro Val Ser Val Phe Trp Lys Thr Val

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Ser Arg Arg Phe Ala Glu Ala Ala Cys Asp Val Val His Val Met Leu

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Asn Gly Ser Arg Ser Lys Ile Phe Asp Lys Asn Ser Thr Phe Gly Ser

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Val Glu Val His Asn Leu Gln Pro Glu Lys Val Gln Thr Leu Glu Ala

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Trp Val Ile His Gly Gly Arg Glu Asp Ser Arg Asp Leu Cys Gln Asp

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Pro Thr Ile Lys Glu Leu Glu Ser Ile Ile Ser Lys Arg Asn Ile Gln

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Phe Ser Cys Lys Asn Ile Tyr Arg Pro Asp Lys Phe Leu Gln Cys Val

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Met Ala Asn Cys Glu Phe Ser Pro Val Ser Gly Asp Lys Pro Cys Cys

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Arg Leu Ser Arg Arg Ala Gln Val Cys Leu Gly Val Cys Leu Leu Val

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Leu Leu Ile Leu Val Val Val Val Ala Val Val Leu Pro Arg Trp Arg

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Gln Gln Trp Ser Gly Ser Gly Thr Thr Ser Arg Phe Pro Glu Thr Val

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Leu Ala Arg Cys Val Lys Tyr Thr Glu Val His Pro Glu Met Arg His

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Val Asp Cys Gln Ser Val Trp Asp Ala Phe Lys Gly Ala Phe Ile Ser

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Lys Tyr Pro Cys Asn Ile Thr Glu Glu Asp Tyr Gln Pro Leu Val Lys

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Leu Gly Thr Gln Thr Val Pro Cys Asn Lys Thr Leu Leu Trp Ser Arg

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Ile Lys Asp Leu Ala His Gln Phe Thr Gln Val Gln Arg Asp Met Phe

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Thr Leu Glu Asp Met Leu Leu Gly Tyr Leu Ala Asp Asp Leu Thr Trp

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Cys Gly Glu Phe Asn Thr Phe Glu Ile Asn Tyr Gln Ser Cys Pro Asp

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Trp Arg Lys Asp Cys Ser Asn Asn Pro Val Ser Val Phe Trp Lys Thr

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Val Ser Arg Arg Phe Ala Glu Thr Ala Cys Gly Val Val His Val Met

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Leu Asn Gly Ser Arg Ser Lys Ile Phe Asp Lys Asn Ser Thr Phe Gly

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Ser Val Glu Val His Asn Leu Gln Pro Glu Lys Val Gln Ala Leu Glu

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Ala Trp Val Ile His Gly Gly Arg Glu Asp Ser Arg Asp Leu Cys Gln

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Asp Pro Thr Ile Lys Glu Leu Glu Ser Ile Ile Ser Lys Arg Asn Ile

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Arg Phe Phe Cys Lys Asn Ile Tyr Arg Pro Asp Lys Phe Leu Gln Cys

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Val Lys Asn Pro Glu Asp Ser Ser Cys Leu Ser Gly Ile

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Gly Phe Thr Phe Asp Asp Tyr Gly

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Ile Ser Trp Asn Gly Gly Lys Thr

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Ser Ser Asn Ile Gly Asp Asn Tyr

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Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

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Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

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Ser Asp Ile Ser Trp Asn Gly Gly Lys Thr His Tyr Val Asp Ser Val

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Lys Gly Gln Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

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Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

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Ala Arg Gly Ser Leu Phe His Asp Ser Ser Gly Phe Tyr Phe Gly His

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Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly

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Pro Ser Val Phe Pro Leu Ala

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<223> amino acid chain sequence (VL)

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Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

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Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asp Asn

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Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Arg Asp Ser Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

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Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg

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Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser Leu

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Ser Gly Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln

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Pro Lys Ala Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser Glu Glu

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Leu

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<223> heavy chain (HC) amino acid sequence

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Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

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Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Asp Ile Ser Trp Asn Gly Gly Lys Thr His Tyr Val Asp Ser Val

50 55 60

Lys Gly Gln Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

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Ala Arg Gly Ser Leu Phe His Asp Ser Ser Gly Phe Tyr Phe Gly His

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Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly

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Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly

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Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

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Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

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Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

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Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val

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Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys

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Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu

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Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

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Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

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Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

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Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

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Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

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Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

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Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

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Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

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Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

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Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

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Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

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Leu Ser Pro Gly Lys

450

<210> 12

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<212> Protein

<213> Artificial sequence

<220>

<223> light chain (LC) amino acid sequence

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Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

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Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asp Asn

20 25 30

Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Arg Asp Ser Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg

65 70 75 80

Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser Leu

85 90 95

Ser Gly Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln

100 105 110

Pro Lys Ala Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser Glu Glu

115 120 125

Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr

130 135 140

Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro Val Lys

145 150 155 160

Ala Gly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn Lys Tyr

165 170 175

Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His

180 185 190

Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys

195 200 205

Thr Val Ala Pro Thr Glu Cys Ser

210 215

<210> 13

<211> 318

<212> Protein

<213> Artificial sequence

<220>

<223> human CD157

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Met Ala Ala Gln Gly Cys Ala Ala Ser Arg Leu Leu Gln Leu Leu Leu

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Gln Leu Leu Leu Leu Leu Leu Leu Leu Ala Ala Gly Gly Ala Arg Ala

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Arg Trp Arg Gly Glu Gly Thr Ser Ala His Leu Arg Asp Ile Phe Leu

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Gly Arg Cys Ala Glu Tyr Arg Ala Leu Leu Ser Pro Glu Gln Arg Asn

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Lys Asn Cys Thr Ala Ile Trp Glu Ala Phe Lys Val Ala Leu Asp Lys

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Asp Pro Cys Ser Val Leu Pro Ser Asp Tyr Asp Leu Phe Ile Asn Leu

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Ser Arg His Ser Ile Pro Arg Asp Lys Ser Leu Phe Trp Glu Asn Ser

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His Leu Leu Val Asn Ser Phe Ala Asp Asn Thr Arg Arg Phe Met Pro

115 120 125

Leu Ser Asp Val Leu Tyr Gly Arg Val Ala Asp Phe Leu Ser Trp Cys

130 135 140

Arg Gln Lys Asn Asp Ser Gly Leu Asp Tyr Gln Ser Cys Pro Thr Ser

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Glu Asp Cys Glu Asn Asn Pro Val Asp Ser Phe Trp Lys Arg Ala Ser

165 170 175

Ile Gln Tyr Ser Lys Asp Ser Ser Gly Val Ile His Val Met Leu Asn

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Gly Ser Glu Pro Thr Gly Ala Tyr Pro Ile Lys Gly Phe Phe Ala Asp

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Tyr Glu Ile Pro Asn Leu Gln Lys Glu Lys Ile Thr Arg Ile Glu Ile

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Trp Val Met His Glu Ile Gly Gly Pro Asn Val Glu Ser Cys Gly Glu

225 230 235 240

Gly Ser Met Lys Val Leu Glu Lys Arg Leu Lys Asp Met Gly Phe Gln

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Tyr Ser Cys Ile Asn Asp Tyr Arg Pro Val Lys Leu Leu Gln Cys Val

260 265 270

Asp His Ser Thr His Pro Asp Cys Ala Leu Lys Ser Ala Ala Ala Ala

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Thr Gln Arg Lys Ala Pro Ser Leu Tyr Thr Glu Gln Arg Ala Gly Leu

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Ile Ile Pro Leu Phe Leu Val Leu Ala Ser Arg Thr Gln Leu

305 310 315

<---

Claims (16)

1. A method of treating relapsed/refractory multiple myeloma in a subject, comprising subcutaneously administering to the subject an isolated human anti-CD38 antibody in sufficient amount, wherein the anti-CD38 antibody comprises a heavy chain variable region (VH) containing CDR1 having the amino acid sequence of SEQ ID NO: 3 , CDR2 having the amino acid sequence of SEQ ID NO: 4, and CDR3 having the amino acid sequence of SEQ ID NO: 5; and a light chain variable region (VL) CDR1 having the amino acid sequence of SEQ ID NO: 6, CDR2 having the amino acid sequence of SEQ ID NO: 7, and CDR3 having the amino acid sequence of SEQ ID NO: 8, wherein the antibody is administered at a dose of 45 up to 1800 mg, where the dose is a once-weekly dose. 2. The method according to claim 1, in which the administration of an antibody to CD38 does not cause hemolytic anemia or thrombocytopenia. 3. The method of any one of the preceding claims, wherein administration of the anti-CD38 antibody results in less than 30% of grade 3 or 4 incidence of one or more treatment-emergent adverse events (TEAEs) or treatment-emergent adverse events (TEAEs) , selected from the group consisting of anemia, hemolytic anemia, thrombocytopenia, fatigue, infusion-related reactions (IRR), leukopenia and lymphopenia. 4. A method as claimed in any one of the preceding claims, wherein the anti-CD38 antibody causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1% depletion of the red blood cell population. 5. A method as claimed in any one of the preceding claims, wherein the anti-CD38 antibody causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1% platelet depletion. 6. The method of any one of the preceding claims, wherein the VH chain region has the amino acid sequence of SEQ ID NO: 9 and the VL chain region has the amino acid sequence of SEQ ID NO: 10. 7. The method of any one of the preceding claims, wherein the anti-CD38 antibody comprises the heavy chain amino acid sequence of SEQ ID NO: 11 and the light chain amino acid sequence of SEQ ID NO: 12. 8. A method as claimed in any one of the preceding claims, wherein the antibody is administered at a dose of 135 to 1800 mg, 600 to 1800 mg, 1200 to 1800 mg, 45 to 1200 mg, 45 to 600 mg, 45 to 135 mg , 135 to 1200 mg, 135 to 600 mg, or 1200 to 1800 mg. 9. The method of any one of the preceding claims, wherein the human anti-CD38 antibody is administered in the form of a pharmaceutically acceptable composition. 10. A unit dosage form for the treatment of relapsed/refractory multiple myeloma, comprising an isolated antibody that contains a heavy chain variable region comprising SEQ ID NO: 9 and a light chain variable region comprising SEQ ID NO: 10, wherein the isolated antibody binds to CD38 and does not bind to human red blood cells and the unit dosage form contains an isolated antibody in an amount of from 45 to 1800 mg, wherein the unit dosage form contains one or more pharmaceutically acceptable excipients, carriers and/or diluents, and wherein the volume of the unit dosage form is 2. 5 ml or less. 11. The unit dosage form of claim 10, wherein the unit dosage form contains the isolated antibody in an amount of 135 to 1800 mg, 600 to 1800 mg, 1200 to 1800 mg, 45 to 1200 mg, 45 to 600 mg, 45 to 135 mg, 135 to 1200 mg, 135 to 600 mg, or 1200 to 1800 mg. 12. The unit dosage form of claim 10 or 11, wherein the isolated antibody comprises a heavy chain comprising SEQ ID NO: 11 and a light chain comprising SEQ ID NO: 12. 13. Unit dosage form according to any one of paragraphs. 10 or 11, in which the anti-CD38 antibody does not cause hemolytic anemia or thrombocytopenia. 14. Unit dosage form according to any one of paragraphs. 10-13, in which the anti-CD38 antibody causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% depletion red blood cell populations. 15. Unit dosage form according to any one of paragraphs. 10-14, in which the anti-CD38 antibody causes less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% depletion platelet populations. 16. Unit dosage form according to any one of paragraphs. 10-15, in which the dosage is a once-weekly dosage.

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