Anti CD19 therapy in patients with limited numbers of NK cells

Abstract

To provide anti- CD19 antibodies for use in treating patients with hematological cancers, and to provide methods for selecting patients with hematological cancers who are predicted to benefit from therapeutically administering anti- CD19 antibodies.SOLUTION: The present invention provides features and biomarkers in patients who benefit from treatment with anti- CD19 antibodies (MOR00208, XmAb5574). In addition, the application provides anti- 100NK antibodies for the treatment of leukemias or lymphomas in patients with peripheral NK cell counts equal to or less than CD19 cells / μ l at baseline.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present disclosure is directed to identifying characteristics and biomarkers in patients who would benefit from treatment with anti-CD19 antibodies. Additionally, the present disclosure relates to anti-CD19 antibodies for the treatment of leukemia or lymphoma in patients with limited numbers of NK cells. [Background technology]

[0002] CD19 is a 95-kDa transmembrane glycoprotein of the immunoglobulin superfamily containing two extracellular immunoglobulin-like domains and an extensive cytoplasmic tail. The protein is a pan-B lymphocyte surface receptor and is ubiquitously expressed from early stages of pre-B cell development until it is downregulated during terminal differentiation into plasma cells. CD19 is B lymphocyte lineage-specific and is not expressed on hematopoietic stem cells or other immune cells, except for some follicular dendritic cells. CD19 functions as a positive regulator of B cell receptor (BCR) signaling and is important for B cell activation and proliferation and the development of humoral immune responses. CD19 acts as a costimulatory molecule in conjunction with CD21 and CD81 and is important for B cell responses to T cell-dependent antigens. The cytoplasmic tail of CD19 physically associates with a family of tyrosine kinases that trigger downstream signaling pathways via the src family of protein tyrosine kinases. CD19 is an attractive target for cancers of lymphatic origin because it is highly expressed in nearly all chronic lymphocytic leukemias (CLL) and non-Hodgkin's lymphomas (NHL), as well as many other different types of leukemia, including acute lymphocytic leukemia (ALL) and hairy cell leukemia (HCL).

[0003] MOR00208 (formerly XmAb® 5574) is a humanized monoclonal antibody targeting the antigen CD19, a transmembrane protein involved in B-cell receptor signaling. MOR00208 has been engineered in the IgG Fc region to enhance antibody-dependent cell-mediated cytotoxicity (ADCC), thus improving an important mechanism for tumor cell killing and offering the potential for enhanced efficacy compared to conventional, i.e., non-enhanced, antibodies. MOR00208 has been or is currently being studied in several clinical trials, including for CLL, ALL, and NHL. In some of these trials, MOR00208 is used in combination with idelalisib, lenalidomide, or venetoclax.

[0004] In a Phase II / III trial called B-MIND, the efficacy and safety of MOR00208 in combination with bendamustine (BEN) will be evaluated in adult patients with relapsed or refractory diffuse large B-cell lymphoma (rrDLBCL). The study will compare the MOR00208+BEN combination with rituximab (RTX) and BEN. The chimeric murine / human antibody rituximab was first approved by the U.S. Food and Drug Administration (FDA) in 1997 for the treatment of patients with relapsed or refractory low-grade or follicular CD20-positive B-cell non-Hodgkin's lymphoma (NHL). In Europe, rituximab was approved for the treatment of NHL patients in 1998.

[0005] Recently, the number of treatment options for patients with B-cell malignancies has increased, and the clinical effectiveness of monoclonal antibodies (mAbs) and mAb-based therapies has been demonstrated in a number of hematological malignancies, primarily in combination with chemotherapeutic agents. However, a significant number of patients with B-cell malignancies are refractory or relapse after initial tumor remission in response to these combined antibody chemotherapies. Overall, variable patient response rates to antibody therapy are observed based on different patient profiles. To further optimize the success of therapy, additional methods are needed to accurately predict which patients are likely to respond and / or are likely to respond best to such antibody therapy. Specific biomarkers or patient characteristics may be found to correlate with responsiveness to such therapy, with specific concentrations or ranges of each biomarker.

[0006] Thus, there is a need for methods of using biomarkers or specific patient characteristics for use in connection with the treatment of cancer with therapies including anti-CD19 antibodies. Summary of the Invention

[0007] The impact of natural killer (NK) cell count (NKCC) on survival in patients with DLBCL treated with rituximab, cyclophosphamide, doxorubicin hydrochloride (hydroxydaunomycin), vincristine sulfate (Oncovin), and prednisone (collectively known as "R-CHOP") was evaluated in Kim et al., Blood Research, 49:3, 162-169 (September 2014). Previously, peripheral NK cell count was reported to be associated with clinical outcome in patients with aaIPI 2-3 DLBCL (Plonquet et al., Ann Oncol 2007;18:1209-15).

[0008] To date, emerging evidence suggests that baseline NKCC has prognostic value in the treatment of B-cell lymphoma with anti-CD20-containing regimens (He et al., Blood Cancer J. 2016 Aug;6(8), Kim et al., Blood Res. 2014 Sep;49(3):162-9, Klanova et al., Blood 2017 130:727). In a large study of over 2,000 patients with previously untreated FL and DLBCL (GALLIUM, GOYA), baseline NKCC was shown to be an independent prognostic parameter by multivariate analysis. 高 Patients with NKCC 低 The cutoff was chosen based on the highest differential effect between both subgroups and was consistently within the range of 100 NK cells per μL of blood. 高 A positive prognosis for patients (more than 100 NK cells per μL of blood) was disclosed in WO2017 / 207574, and in the MOR00208C201 study a cutoff of at least 100 NK cells / μL was used as prognostic for treatment outcome of MOR00208 monotherapy in DLBCL and FL.

[0009] However, NKCC patients with blood cancer 低 Therefore, these patients are considered to have a poor prognosis based on their low NKCC, which translates into a specific high unmet medical need for this particular patient subgroup.

[0010] The present disclosure provides a method for treating NKCC in patients with B-cell malignancies, such as non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), and / or acute lymphoblastic leukemia (ALL). 低 The present disclosure relates to improved methods for the treatment of patients, particularly those suffering from NKCC. 低Antibodies specific for CD19 for the treatment of B-cell malignancies such as non-Hodgkin's lymphoma, chronic lymphocytic leukemia and / or acute lymphoblastic leukemia in patients.

[0011] In this disclosure, the ADCC activity of MOR00208 was compared with that of the anti-CD20 antibody rituximab in B-cell tumor cell lines at various effector-to-target (E:T) ratios. Rituximab can be described as the gold standard treatment for these indications. The target cell lines were derived from DLBCL, MCL, and CLL, and CD19 and CD20 levels were within the range of expression levels reported on B-cell tumor patient samples by Boltezar et al. (2018), Ginaldi et al. (1998), and Olejniczak et al. (2006). Using the respective E:T ratios in ADCC assays for several cell lines, potential correlations between the advantages of MOR00208 versus rituximab and MOR00208 versus NKCC were elucidated. The resulting data demonstrated an increasing relative benefit of MOR00208 with decreasing E:T ratios, thus demonstrating a significant benefit for NKCC. 低 We provided a rationale for the superiority of MOR00208 in the subgroups. Based on the available data for determining the cutoff, NKCC 低 A subgroup is defined as patients with 100 NK cells / μl or less at baseline.

[0012] Therefore, patients diagnosed with a B-cell malignancy, such as non-Hodgkin's lymphoma, chronic lymphocytic leukemia and / or acute lymphoblastic leukemia, and who have a baseline peripheral NK cell count of 100 cells / μl or less at baseline, are more likely to benefit from MOR00208 treatment compared to available therapies.

[0013] The present disclosure provides anti-CD19 antibodies for use in treating patients with hematological cancers, the patients having a baseline peripheral NK cell count of 100 cells / μl or less. In one embodiment, the patient is resistant, non-responsive, or inadequately responsive to treatment with no more than one to three prior lines of therapy, including one anti-CD20 targeted therapy (e.g., the antibody rituximab). In a further embodiment, the patient is not a candidate for high-dose chemotherapy and autologous stem cell transplantation. In a preferred embodiment, the patient is human.

[0014] In embodiments, an anti-CD19 antibody for use in treating patients with hematological cancer comprises an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6).

[0015] In a further embodiment, the anti-CD19 antibody for use in treating patients with hematological cancer comprises the sequence The variable heavy chain of EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7) and array The variable light chain of the compound of formula (SEQ ID NO: 8) is: DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK.

[0016] In another embodiment of the present disclosure, the anti-CD19 antibody is a human, humanized, or chimeric antibody. In another embodiment of the present disclosure, the anti-CD19 antibody is of the IgG isotype. In another embodiment, the antibody is an IgG1, IgG2, or IgG1 / IgG2 chimera. In another embodiment of the present disclosure, the isotype of the anti-CD19 antibody is engineered to enhance antibody-dependent cell-mediated cytotoxicity. In another embodiment, the heavy chain constant region of the anti-CD19 antibody comprises amino acids 239D and 332E, and Fc numbering is according to the EU index as in Kabat. In another embodiment, the antibody is an IgG1, IgG2, or IgG1 / IgG2, and the chimeric heavy chain constant region of the anti-CD19 antibody comprises amino acids 239D and 332E, and Fc numbering is according to the EU index as in Kabat.

[0017] In a further embodiment, the anti-CD19 antibody for use in treating patients with hematological cancer comprises the sequence EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP A heavy chain having CPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11) and array and a light chain having the sequence DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12). [Brief explanation of the drawings]

[0018] [Figure 1] Representative ADCC assay of MOR00208 and rituximab at increasing E:T ratios. Results of specific killing, expressed as % killed target cells, mediated by MOR00208 (black) or rituximab (white) and NK cells from healthy donors are shown for three target cell lines: MEC-1, JVM-2, and Toledo. Exemplary data obtained with NK cells from one representative donor in a single experiment are shown. Bars and error bars represent the geometric mean and geometric standard deviation of specific killing measured in triplicate from individual experiments. [Figure 2] Representative ADCC assay of MOR00208 and rituximab at increasing E:T ratios. The corresponding ratios of specific killing mediated by MOR00208 (blue) or rituximab (orange) and NK cells from healthy donors, normalized to rituximab, are shown for three target cell lines: MEC-1, JVM-2, and Toledo. Scatter plots show the individual values ​​of the specific killing ratios of MOR00208 (triangles) and rituximab (circles) versus the median rituximab value of a representative experiment. The dotted line represents the geometric mean with a 95% bootstrap confidence interval. [Figure 3]Specific killing ratios of MOR00208 and rituximab in ADCC assays at increasing E:T ratios using NK cells isolated from 33 healthy donors, as well as MEC-1, JVM-2, and Toledo cells. The ADCC activity of MOR00208 (open triangles) and rituximab (black circles) was analyzed in MEC-1 cells (8 NK cell donors, two independent experiments; 9 NK cell donors in a single experiment), JVM-2 (8 NK cell donors, two independent experiments per donor), and Toledo cells (10 NK cell donors, two independent experiments). Specific killing ratios were calculated from the % specific killing determined for each antibody by normalizing to the median value for rituximab. Each circle or triangle represents the geometric mean value of one or two independent experiments performed in triplicate using NK cells from one individual blood donor. DETAILED DESCRIPTION OF THE INVENTION

[0019] Non-Hodgkin's lymphoma (NHL) is a heterogeneous malignant tumor originating from lymphocytes. In the United States (US), the incidence rate is estimated at 65,000 per year, with approximately 20,000 deaths (American Cancer Society, 2006 and SEER Cancer Statistics Review). The disease can occur at any age, with onset typically beginning in adults over 40 years of age, and incidence rates increasing with age. NHL is characterized by the clonal proliferation of lymphocytes that accumulate in lymph nodes, blood, bone marrow, and spleen, although any major organ may be involved. The current classification system used by pathologists and clinicians is the World Health Organization (WHO) Classification of Tumors, which organizes NHL into precursor and mature B-cell or T-cell neoplasms. PDQ currently divides NHL into indolent or aggressive for the purpose of clinical trial enrollment. The indolent NHL group is primarily composed of follicular subtypes, small lymphocytic lymphoma, mucosa-associated lymphoid tissue (MALT), and marginal zone lymphoma, and indolent NHL encompasses approximately 50% of newly diagnosed B-cell NHL patients. Aggressive NHL primarily includes patients with diffuse large B-cell (DLBL, "DLBCL," or DLCL) histologic diagnoses (40% of all newly diagnosed patients have diffuse large cell), Burkitt cell, and mantle cell ("MCL") histologic diagnoses. The clinical course of NHL is highly variable. The primary determinant of clinical course is the histologic subtype. Most indolent NHL is considered incurable. Patients initially respond to either chemotherapy or antibody therapy, and most relapse. Previous studies have not demonstrated improved survival with early intervention. In asymptomatic patients, "waiting" is acceptable until the patient develops symptoms or the pace of the disease appears to be accelerating. Over time, the disease may evolve to a more aggressive histology. Median survival is 8-10 years, and indolent patients often undergo three or more lines of therapy during their treatment phase. Initial treatment for symptomatic indolent NHL has historically been combination chemotherapy.The most commonly used drugs include cyclophosphamide, vincristine, and prednisone (CVP) or cyclophosphamide, adriamycin, vincristine, and prednisone (CHOP). Approximately 70% to 80% of patients respond to initial chemotherapy, with remissions lasting 2 to 3 years. Ultimately, the majority of patients relapse. The discovery and clinical use of the anti-CD20 antibody rituximab has significantly improved response rates and survival rates. The current standard of care for most patients is rituximab plus CHOP (R-CHOP) or rituximab plus CVP (R-CVP). Rituximab therapy has been shown to be effective in several types of NHL and is currently approved as first-line treatment for both indolent (follicular lymphoma) and aggressive (diffuse large B-cell lymphoma) NHL. However, anti-CD20 monoclonal antibodies (mAbs) have significant limitations, including primary resistance (50% response in slow-relapse patients), acquired resistance (50% response rate upon retreatment), rare complete responses (2% complete response rate in the relapse population), and a continuous pattern of relapse. Finally, many B-cell disorders cannot be treated using anti-CD20 antibody therapy because many B cells do not express CD20.

[0020] In addition to NHL, there are several types of leukemia resulting from dysregulation of B cells. Chronic lymphocytic leukemia (also known as "chronic lymphocytic leukemia" or "CLL") is a type of adult leukemia caused by the abnormal accumulation of B lymphocytes. In CLL, malignant lymphocytes may appear normal and mature but are unable to effectively respond to infection. CLL is the most common form of leukemia in adults. Men are twice as likely as women to develop CLL. However, age is a significant risk factor. More than 75% of new cases are diagnosed in patients over the age of 50. More than 10,000 cases are diagnosed each year, with nearly 5,000 deaths annually (American Cancer Society, 2006 and SEER Cancer Statistics Review). CLL is incurable, but the disease progresses slowly in most cases. Many CLL patients lead normal, active lives for many years. Because of the slow onset of the disease, early intervention is not believed to improve survival or quality of life, so early-stage CLL is generally not treated. Instead, the condition is monitored over time. Initial CLL treatment varies depending on the exact diagnosis and progression of the disease. Dozens of drugs are used in CLL therapy. Combination chemotherapy regimens such as FCR (fludarabine, cyclophosphamide, and rituximab) and BR (ibrutinib and rituximab) are effective in both newly diagnosed and relapsed CLL. Allogeneic bone marrow (stem cell) transplantation is rarely used as a first-line treatment for CLL due to its risks.

[0021] Another type of leukemia is small lymphocytic lymphoma ("SLL"), which lacks the clonal lymphocytosis required for the diagnosis of CLL but is otherwise considered a CLL variant that shares pathological and immunophenotypic features (Campo et al., 2011). The definition of SLL requires the presence of lymphadenopathy and / or splenomegaly. Furthermore, the number of B lymphocytes in the peripheral blood should not exceed 5 × 109 / L. Whenever possible, SLL diagnosis should be confirmed by histopathological evaluation of lymph node biopsies (Hallek et al., 2008). The incidence of SLL is approximately 25% of CLL cases in the United States (Dores et al., 2007).

[0022] Another type of leukemia is acute lymphoblastic leukemia (ALL), also known as acute lymphocytic leukemia. ALL is characterized by the overproduction and continuous proliferation of malignant, immature white blood cells (also known as lymphoblasts) in the bone marrow. The "acute" term refers to the undifferentiated, immature state of circulating lymphocytes ("blasts"). If left untreated, the disease progresses rapidly, with an average lifespan of several weeks to several months. ALL is most common in childhood, with a peak incidence between the ages of 4 and 5. Children aged 12 to 16 are more likely to die than other children. Currently, at least 80% of childhood ALL cases are considered curable. Fewer than 4,000 cases are diagnosed each year, with approximately 1,500 deaths annually (American Cancer Society, 2006 and SEER Cancer Statistics Review).

[0023] The use of CD19 antibodies in non-specific B-cell lymphoma is discussed in WO2007 / 076950 (US2007 / 154473), both of which are incorporated by reference. The use of CD19 antibodies in CLL, NHL, and ALL is described in Scheuermann et al., CD19 Antigen in Leukemia and Lymphoma Diagnosis and Immunotherapy, Leukemia and Lymphoma, Vol. 18, 385-397 (1995), which is incorporated by reference in its entirety.

[0024] Further antibodies specific for CD19 are disclosed in WO2005 / 012493 (US7109304), WO2010 / 053716 (US12 / 266,999) (Immunomedics); WO2007 / 002223 (US8097703) (Medarex); WO2008 / 022152 (US12 / 377,251) and WO2008 / 150494 (Xencor), WO2008 / 031056 (US11 / 852,106) (Medimmune); WO2007 / 076950 (US11 / 648,505) (Merck Patent GmbH); WO2009 / 052431 (US12 / 253,895) (Seattle Genetics); and WO2010 / 095031(12 / 710,442) (Glenmark Pharmaceuticals), WO2012 / 010562 and WO2012 / 010561 (International Drug Development), WO2011 / 147834 (Roche Glycart), and WO2012 / 156455 (Sanofi), all of which are incorporated by reference in their entireties.

[0025] The pharmaceutical composition comprises an active agent, such as an antibody, for therapeutic use in humans. The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier or excipient.

[0026] definition The term "CD19" refers to the protein known as CD19, and has the following synonyms: B4, B lymphocyte antigen CD19, B lymphocyte surface antigen B4, CVID3, differentiation antigen CD19, MGC12802, and T cell surface antigen Leu-12.

[0027] Human CD19 has the following amino acid sequence: (SEQ ID NO: 13)

[0028] "MOR00208" is an anti-CD19 antibody. The amino acid sequence is provided in Table 1. "MOR00208" and "XmAb 5574" are used synonymously to describe the antibody shown in Table 1. The MOR00208 antibody is described in U.S. Patent Application No. 12 / 377,251, which is incorporated by reference in its entirety. U.S. Patent Application No. 12 / 377,251 describes an antibody called 4G7 H1.52 hybrid S239D / I332E / 4G7 L1.155 (later named MOR00208).

[0029] As used herein, the term "antibody" refers to a protein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds that interact with an antigen. Each heavy chain is composed of a variable heavy chain region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region is composed of three domains, CH1, CH2, and CH3. Each light chain is composed of a variable light chain region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is composed of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability called complementarity-determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The term "antibody" includes, for example, monoclonal antibodies, human antibodies, humanized antibodies, camelized antibodies, and chimeric antibodies. Antibodies can be of any isotype (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. Both the light and heavy chains are divided into regions of structural and functional homology.

[0030] The term "antibody fragment" as used herein refers to one or more portions of an antibody that retain the ability to specifically interact with an antigen (e.g., by binding, steric hindrance, or stabilization of spatial distribution). Examples of binding fragments include, but are not limited to, a Fab fragment, which is a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; a F(ab)2 fragment, which is a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; an Fd fragment consisting of the VH and CH1 domains; an Fv fragment consisting of the VL and VH domains of a single antibody arm; a dAb fragment consisting of the VH domain (Ward et al., (1989) Nature 341:544-546); and isolated complementarity-determining regions (CDRs). Furthermore, although the two domains of an Fv fragment, VL and VH, are encoded by separate genes, they can be joined by a synthetic linker that allows them to be produced using recombinant methods as a single protein chain in which the VL and VH regions pair to form a monovalent molecule (known as a single-chain Fv (scFv); see, e.g., Bird et al., (1988) Science 242:423-426 and Huston et al., (1988) Proc. Natl. Acad. Sci. 85:5879-5883). Such single-chain antibodies are also intended to be encompassed within the term "antibody fragment." These antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for utility in the same manner as intact antibodies. Antibody fragments can also be incorporated into single domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NARs, and bis-scFvs (see, e.g., Hollinger and Hudson, (2005) Nature Biotechnology 23:1126-1136). Antibody fragments can be grafted onto scaffolds based on polypeptides such as fibronectin type III (Fn3) (see U.S. Pat. No. 6,703,199, which describes fibronectin polypeptide monobodies).Antibody fragments can be assembled into single-chain molecules comprising a pair of tandem Fv segments (VH-CH1-VH-CH1) which, together with complementary light chain polypeptides, form a pair of antigen-binding sites (Zapata et al., (1995) Protein Eng. 8:1057-1062 and U.S. Pat. No. 5,641,870).

[0031] "Administered" or "administration" includes, but is not limited to, delivery of a drug by an injectable form, such as, for example, intravenous, intramuscular, intradermal or subcutaneous routes, or mucosal routes, for example, as a nasal spray or aerosol for inhalation, or as an ingestible solution, capsule or tablet. Preferably, administration is by an injectable form.

[0032] The term "effector function" refers to biological activities attributable to the Fc region of an antibody, which vary depending on the antibody isotype. Non-limiting examples of antibody effector functions include C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding and antibody-dependent cell-mediated cytotoxicity (ADCC) and / or antibody-dependent cellular phagocytosis (ADCP), down-regulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0033] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which antibodies bound to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., NK cells, neutrophils, and macrophages) enable these cytotoxic effector cells to specifically bind to antigen-bearing target cells and then kill the target cells with cytotoxins. NK cells, the primary cells for mediating ADCC, express only FcγRIII, whereas monocytes express FcγRI, FcγRII, and FcγRIII.

[0034] "Complement-dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to antibodies (of the appropriate subclass) of the present disclosure that are bound to their cognate antigen.

[0035] "Antibody-dependent cellular phagocytosis" or "ADCP" refers to the mechanism of elimination of antibody-coated target cells by internalization by phagocytic cells such as macrophages or dendritic cells.

[0036] The term "hematological cancer" includes blood-borne tumors and diseases or disorders involving abnormal cell growth and / or proliferation in tissues of hematopoietic origin, such as lymphoma, leukemia, and myeloma.

[0037] As used in this context, a "subject" or "patient" refers to any mammal, including rodents such as mice or rats, as well as primates such as cynomolgus monkeys (Macaca fascicularis), rhesus monkeys (Macaca mulatta), or humans (Homo sapiens). Preferably, the subject or patient is a primate, most preferably a human.

[0038] As used herein, the terms "engineered" or "modified" include the manipulation of nucleic acids or polypeptides by synthetic means (e.g., recombinant techniques, in vitro peptide synthesis, enzymatic or chemical coupling of peptides, or some combination of these techniques). Preferably, antibodies or antibody fragments according to the present disclosure are engineered or modified to improve one or more properties, e.g., antigen binding, stability, half-life, effector function, immunogenicity, safety, etc. Preferably, antibodies or antibody fragments according to the present disclosure are engineered or modified to improve effector function, such as ADCC.

[0039] As used herein, a "variant" refers to a polypeptide that differs from a reference polypeptide by one or more modifications, eg, an amino acid substitution, insertion, or deletion.

[0040] As used herein, the term "antagonist" antibody refers to an antibody or antibody fragment that interacts with an antigen and partially or completely inhibits or neutralizes the biological activity or function, or any other phenotypic characteristic, of the target antigen.

[0041] "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain. The Fc region of an immunoglobulin generally comprises two constant domains, a CH2 domain and a CH3 domain. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region is in accordance with Kabat et al., Sequences of Proteins of Immunological Interest, 5 th The EU numbering system, also known as the EU index, is followed as described in Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0042] Antibodies administered in accordance with the present disclosure are administered to patients in therapeutically effective amounts. A "therapeutically effective amount" refers to an amount sufficient to provide some improvement in the clinical symptoms of a given disease or disorder. The amount effective for a particular therapeutic purpose will depend on the severity of the disease or injury, as well as the weight and general condition of the subject. It will be understood that determining the appropriate dosage can be accomplished using routine experimentation, constructing a matrix of values, and testing different points within the matrix, all of which is within the ordinary skill of a trained physician or clinical scientist.

[0043] "Baseline" or "at baseline" means prior to administration of the desired therapy, e.g., prior to administration of the desired anti-CD19 antibody.

[0044] Receiver operating characteristic (ROC) analysis can be used to analyze the predictive power, sensitivity, and specificity of potential biomarkers, such as NK cell counts, to determine their cutoff values. Additional methods exist for estimating optimal cutoff values: a) "Max.Accuracy"—the cutoff value that maximizes accuracy; b) "Max.DOR"—the cutoff value that maximizes the diagnostic odds ratio; c) "Error.rate"—the cutoff value that minimizes the error rate; d) "Max.Accuracy.area"—the cutoff value that maximizes the accuracy area; e) "Max.Sens+Spec"—the cutoff value that maximizes the sum of sensitivity and specificity; f) "Max.Youden"—the cutoff value that maximizes the Youden index; g) "Se=Sp"—the cutoff value where sensitivity equals specificity; h) "Min.ROC.Dist"—the cutoff value that minimizes the distance between the curve and the upper left corner of the graph; i) "Max.Efficiency"—the cutoff value that maximizes efficiency; and j) "Min.MCT"—the cutoff value that minimizes misclassification costs. See Lopez-Raton, M., Rodriguez-Alvarez, MX, Cadarso-Suarez, C. and Gude-Sampedro, F. (2014). Optimal Cutpoints: An R Package for Selecting Optimal Cutpoints in Diagnostic Tests. Journal of Statistical Software 61(8), 1-36.

[0045] CD19-specific antibodies have also been tested preclinically in combination with other drugs, for example, MOR00208 has been tested in combination with nitrogen mustards, purine analogs, thalidomide analogs, phosphoinositide 3-kinase inhibitors, BCL-2 inhibitors, and Bruton's tyrosine kinase (BTK) inhibitors.

[0046] "In combination with" refers to the administration of one therapy in addition to another. Thus, "in combination with" includes simultaneous (e.g., concurrent) and sequential administration in any order. As a non-limiting example, a first therapy (e.g., an agent such as an anti-CD19 antibody) may be administered to a patient at or after (e.g., 1 minute, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 The anti-CD19 antibody and the agent or pharmaceutically acceptable salt thereof may be administered simultaneously or sequentially (e.g., 1 week, 9 weeks, 10 weeks, 11 weeks, or 12 weeks), simultaneously, or sequentially (e.g., 1 minute, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 or more weeks). In some embodiments, the term "combination" means that the anti-CD19 antibody and the agent or pharmaceutically acceptable salt thereof are administered simultaneously or sequentially. In certain embodiments, the anti-CD19 antibody and the agent or pharmaceutically acceptable salt thereof are administered in separate compositions, i.e., the anti-CD19 antibody and the agent or pharmaceutically acceptable salt thereof are each administered in a separate unit dosage form. It will be understood that the anti-CD19 antibody and the agent or pharmaceutically acceptable salt thereof may be administered on the same day or on different days, in any order, according to any appropriate dosing protocol.

[0047] "Nitrogen mustards" are nonspecific DNA alkylating agents used as chemotherapy. Alkylating agents attach alkyl groups (CnH2n+1) to nucleic acid bases, for example, to the guanine base of DNA at the nitrogen atom with atomic number 7 of the imidazole ring. The alkylation step results in the formation of interstrand crosslinks (ICLs). These ICLs are highly cytotoxic because they block essential metabolic processes such as replication and transcription. Nitrogen mustards include cyclophosphamide, chlorambucil, uramustine, ifosfamide, melphalan, and bendamustine.

[0048] Bendamustine is commercially available under the names Ribomustin® and Treanda®, and is also known as SDX-105. Bendamustine is indicated for the treatment of chronic lymphocytic leukemia (CLL), indolent B-cell non-Hodgkin's lymphoma (NHL), and other lymphomas. Bendamustine has the following structure: [ka]

[0049] Purine analogs are antimetabolites that mimic the structure of metabolic purines, thereby interfering with the synthesis of nucleic acids. For example, fludarabine can be incorporated into RNA and DNA by substituting the purine nucleotides adenine and guanine. Purine analogs inhibit the growth of rapidly proliferating cells in an individual, such as cancer cells, bone marrow cells, or cells present in the gastrointestinal tract. Purine analogs include mercaptopurine, azathioprine, thioguanine, and fludarabine. Fludarabine or fludarabine phosphate (Fludara®) is a chemotherapy drug used to treat chronic lymphocytic leukemia and indolent non-Hodgkin's lymphoma. Fludarabine is a purine analog. Fludarabine inhibits DNA synthesis by interfering with ribonucleotide reductase and DNA polymerase, and is S-phase specific (because these enzymes are highly active during DNA replication). Fludarabine has the following structure: [ka]

[0050] "Thalidomide analogs" include, but are not limited to, thalidomide itself, lenalidomide (CC-5013, Revlimid™), pomalidomide (CC4047, Actimid™), and compounds disclosed in WO 2002 / 068414 and WO 2005 / 016326 (incorporated by reference in their entireties). The term refers to synthetic chemical compounds that use the thalidomide structure as a backbone (e.g., side groups have been added or such groups have been deleted from the parent structure). Analogs differ structurally from thalidomide and its metabolite compounds by, for example, differences in alkyl chain length, molecular fragmentation by one or more functional groups, or changes in ionization. The term "thalidomide analogs" also includes metabolites of thalidomide. Thalidomide analogs include racemic mixtures of the S- and R-enantiomers of each compound, as well as the S- or R-enantiomers individually, with the racemic mixtures being preferred.

[0051] Thalidomide analogs include compounds such as lenalidomide, which has the following structure: [ka]

[0052] "Phosphoinositide 3-kinase inhibitors" are a class of drugs that function by inhibiting one or more of the phosphoinositide 3-kinase enzymes that are part of the PI3K / AKT / mTOR pathway, a key signaling pathway for many cellular functions such as growth control, metabolism, and translation initiation.

[0053] There are several different classes and isoforms of PI3K. Class 1 PI3K has a catalytic subunit known as p110 and has four types (isoforms): p110α, p110β, p110γ, and p110δ. Current inhibitors being investigated inhibit one or more isoforms of Class I PI3K.

[0054] Phosphoinositide 3-kinase inhibitors include at least idelalisib, duvelisib, and copanlisib. Idelalisib is marketed by Gilead Sciences, Inc. (trade name Zydelig, also known as GS-1101 or CAL-101). Idelalisib, in combination with rituximab, is currently indicated for the treatment of relapsed chronic lymphocytic leukemia (CLL) in patients for whom rituximab alone is considered appropriate therapy due to other comorbidities, relapsed follicular B-cell non-Hodgkin's lymphoma (FL) in patients who have received at least two prior systemic therapies, and relapsed small lymphocytic lymphoma (SLL) in patients who have received at least two prior systemic therapies. This substance acts as a phosphoinositide 3-kinase inhibitor, more specifically, blocks P110δ, the delta isoform of the enzyme phosphoinositide 3-kinase.

[0055] The formula for idelalisib is: [ka]

[0056] "Bruton's tyrosine kinase (BTK) inhibitors" are a class of drugs that function by inhibiting the tyrosine-protein kinase BTK enzyme, which plays a key role in B cell development. Specifically, BTK contains a PH domain that binds phosphatidylinositol (3,4,5)-trisphosphate (PIP3). PIP3 binding induces Btk to phosphorylate phospholipase C, which then hydrolyzes the phosphatidylinositol, PIP2, into two second messengers, inositol triphosphate (IP3) and diacylglycerol (DAG), which then regulates the activity of downstream proteins during B cell signaling.

[0057] Bruton's tyrosine kinase (BTK) inhibitors include ibrutinib. Ibrutinib is marketed by Pharmacyclics, Inc. and Johnson & Johnson's Janssen Pharmaceutical (trade name Imbruvica, also known as PCI-32765). Ibrutinib is currently indicated for the treatment of patients with mantle cell lymphoma (MCL) who have received at least one prior therapy, chronic lymphocytic leukemia (CLL) who have received at least one prior therapy, chronic lymphocytic leukemia with 17p deletion, and Waldenstrom's macroglobulinemia. The formula of ibrutinib is 1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl]-2-propen-1-one and has the following structure: [ka]

[0058] "BCL-2 inhibitors" are a class of drugs that function by inhibiting the anti-apoptotic B-cell lymphoma-2 (Bcl-2) protein, resulting in programmed cell death of cells. BCL-2 inhibitors include venetoclax, which is marketed by Abbvie and Genentech (under the trade names VENCLEXTA™, also known as GDC-0199, ABT-199, and RG7601). Venetoclax is currently labeled for the treatment of patients with chronic lymphocytic leukemia (CLL) with a 17p deletion detected by an FDA-approved test who have received at least one prior therapy. "Venetoclax" is described in U.S. Patent Nos. 8,546,399 and 9,174,982, all of which are incorporated by reference in their entireties. The formula for venetoclax is: 4-(4-{[2-(4-chlorophenyl)-4,4-dimethyl-1-cyclohexen-1-yl]methyl}-1-piperazinyl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)benzamide, having the following structure: [ka]

[0059] Embodiment In other embodiments, the present disclosure refers to an anti-CD19 antibody for use in treating a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less.

[0060] In other embodiments, the present disclosure refers to an anti-CD19 antibody for use in treating a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of less than 100 cells / μl, less than 90 cells / μl, less than 80 cells / μl, less than 70 cells / μl, less than 60 cells / μl, or less than 50 cells / μl.

[0061] In other embodiments, the disclosure refers to an anti-CD19 antibody for use in treating a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of 1 to up to 100 cells / μl, 10 to up to 100 cells / μl, 20 to up to 100 cells / μl, 30 to up to 100 cells / μl, 40 to up to 100 cells / μl, 50 to up to 100 cells / μl, 60 to up to 100 cells / μl, 70 to up to 100 cells / μl, or 80 to up to 100 cells / μl.

[0062] In other embodiments, the present disclosure refers to the use of an anti-CD19 antibody for use in treating a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of less than 100 cells / μl, less than 90 cells / μl, less than 80 cells / μl, less than 70 cells / μl, less than 60 cells / μl, or less than 50 cells / μl.

[0063] In other embodiments, the present disclosure refers to the use of an anti-CD19 antibody for the treatment of a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of up to 100 cells / μl, up to 90 cells / μl, up to 80 cells / μl, up to 70 cells / μl, up to 60 cells / μl, or up to 50 cells / μl.

[0064] In other embodiments, the disclosure refers to the use of anti-CD19 antibodies for the treatment of patients with hematological cancers, wherein the patients have a baseline peripheral NK cell count of 1 to up to 100 cells / μl, 10 to up to 100 cells / μl, 20 to up to 100 cells / μl, 30 to up to 100 cells / μl, 40 to up to 100 cells / μl, 50 to up to 100 cells / μl, 60 to up to 100 cells / μl, 70 to up to 100 cells / μl, or 80 to up to 100 cells / μl.

[0065] In embodiments, the anti-CD19 antibody for use in treating patients with hematological cancer comprises the sequence The variable heavy chain of EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7) and array The variable light chain of DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK (SEQ ID NO: 8), or a variable heavy chain and a variable light chain having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to the variable heavy chain of SEQ ID NO:7 and the variable light chain of SEQ ID NO:8.

[0066] In embodiments, the anti-CD19 antibody for use in treating patients with hematological cancer comprises the sequence The variable heavy chain of EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7) and array The variable light chain of DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK (SEQ ID NO: 8), or a variable heavy chain and a variable light chain having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the variable heavy chain of SEQ ID NO: 7 and the variable light chain of SEQ ID NO: 8, wherein the anti-CD19 antibody comprises an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6). In another embodiment, the heavy chain region of the anti-CD19 antibody comprises amino acids 239D and 332E, and Fc numbering is according to the EU index as in Kabat.

[0067] In a further embodiment, the anti-CD19 antibody for the treatment of patients with hematological cancer comprises the sequence EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP A heavy chain having CPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11) and array a light chain having DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12); or a heavy chain and a light chain having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to the heavy chain of SEQ ID NO:7 and the light chain of SEQ ID NO:8.

[0068] In a further embodiment, the anti-CD19 antibody for the treatment of patients with hematological cancer comprises the sequence EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP A heavy chain having CPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11) and array a light chain having DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12); or a heavy chain and a light chain having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the heavy chain of SEQ ID NO: 7 and the light chain of SEQ ID NO: 8, wherein the anti-CD19 antibody comprises an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6). In another embodiment, the heavy chain region of the anti-CD19 antibody comprises amino acids 239D and 332E, and Fc numbering is according to the EU index as in Kabat.

[0069] In other embodiments, the present disclosure refers to anti-CD19 antibodies for the treatment of a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less. In one embodiment of the disclosure, the hematological cancer patient after the treatment: (i) a progression-free survival (PFS) of at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 13 months, at least 14 months, at least 15 months, at least 16 months, at least 17 months, at least 18 months, at least 19 months, at least 20 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, at least 48 months, or at least 54 months; (ii) an objective response rate (ORR) of at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 80%; (iii) duration of response (DoR) of at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, at least 48 months, or at least 54 months; (iv) overall survival (OS) of at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, at least 48 months, or at least 54 months; or (v) A combination of one or more of the above. In another embodiment of the disclosure, the anti-CD19 antibody is administered in combination with an agent disclosed herein.

[0070] In other embodiments, the present disclosure refers to anti-CD19 antibodies for the treatment of a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of less than 100 cells / μl, less than 90 cells / μl, less than 80 cells / μl, less than 70 cells / μl, less than 60 cells / μl, or less than 50 cells / μl. In one embodiment of the disclosure, the hematological cancer patient after the treatment: (i) a progression-free survival (PFS) of at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 13 months, at least 14 months, at least 15 months, at least 16 months, at least 17 months, at least 18 months, at least 19 months, at least 20 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, at least 48 months, or at least 54 months; (ii) an objective response rate (ORR) of at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 80%; (iii) duration of response (DoR) of at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, at least 48 months, or at least 54 months; (iv) overall survival (OS) of at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, at least 48 months, or at least 54 months; or (v) A combination of one or more of the above. In another embodiment of the disclosure, the anti-CD19 antibody is administered in combination with an agent disclosed herein.

[0071] In other embodiments, the disclosure is directed to an anti-CD19 antibody for the treatment of a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less, and wherein the anti-CD19 antibody increases one or more of the following characteristics: (i) progression-free survival (PFS), (ii) objective response rate (ORR); (iii) duration of response (DoR); (iv) overall survival (OS); (v) Time to progression (TTP).

[0072] In another embodiment, one or more of the characteristics (i)-(v) are increased compared to a treatment comprising an anti-CD20 antibody. In a further embodiment, one or more of the characteristics (i)-(v) are increased compared to a treatment comprising an anti-CD20 antibody and a chemotherapeutic agent. In a further embodiment, the anti-CD20 antibody is rituximab or a biosimilar thereof. In a further embodiment, one or more of the characteristics (i)-(v) are increased compared to a treatment comprising an anti-CD20 antibody and one or more of cyclophosphamide, adriamycin, vincristine, or prednisone. In a further embodiment, one or more of the characteristics (i)-(v) are increased compared to a treatment comprising R-CHOP.

[0073] In other embodiments, the disclosure refers to an anti-CD19 antibody for the treatment of a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of less than 100 cells / μl, less than 90 cells / μl, less than 80 cells / μl, less than 70 cells / μl, less than 60 cells / μl, or less than 50 cells / μl, and wherein administration of the anti-CD19 antibody results in improved progression-free survival (PFS), improved objective response rate (ORR), improved duration of response (DoR), improved overall survival (OS), or improved time to progression (TTP).

[0074] In other embodiments, the disclosure refers to an anti-CD19 antibody for the treatment of a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less, and wherein administration of the anti-CD19 antibody results in improved progression-free survival (PFS) compared to administration of an anti-CD20 antibody, improved objective response rate (ORR) compared to administration of an anti-CD20 antibody, improved duration of response (DoR) compared to administration of an anti-CD20 antibody, improved overall survival (OS) compared to administration of an anti-CD20 antibody, or improved time to progression-free progression (TTP) compared to administration of an anti-CD20 antibody.

[0075] In other embodiments, the present disclosure refers to an anti-CD19 antibody for the treatment of a patient with a hematological cancer, the patient having a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less, and administration of the anti-CD19 antibody improves progression-free survival (PFS) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent, improves objective response rate (ORR) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent, improves duration of response (DoR) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent, improves overall survival (OS) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent, or improves time to progression (TTP) compared to administration of an anti-CD20 antibody. In a further embodiment, the anti-CD20 antibody is rituximab or a biosimilar thereof. In further embodiments, the chemotherapeutic agents include one or more of cyclophosphamide, adriamycin, vincristine, or prednisone.

[0076] In other embodiments, the disclosure refers to an anti-CD19 antibody for the treatment of a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less, and wherein administration of the anti-CD19 antibody results in improved progression-free survival (PFS) compared to administration of R-CHOP, improved objective response rate (ORR) compared to administration of R-CHOP, improved duration of response (DoR) compared to administration of R-CHOP, improved overall survival (OS) compared to administration of R-CHOP, or improved time to progression (TTP) compared to administration of R-CHOP.

[0077] In one embodiment, the disclosure provides an anti-CD19 antibody, wherein the anti-CD19 antibody is administered at a concentration of 12 mg / kg.

[0078] In a further embodiment, the anti-CD19 antibody is administered weekly, biweekly, or monthly. In a further embodiment, the anti-CD19 antibody is administered weekly for the first three months and then biweekly for at least the next three months. In a further embodiment, the anti-CD19 antibody is administered weekly for the first three months. In a further embodiment, the anti-CD19 antibody is administered weekly for the first three months and then biweekly for at least the next three months. In another embodiment, the anti-CD19 antibody is administered weekly for the first three months, then biweekly for the next three months, and then monthly. In yet another embodiment, the anti-CD19 antibody is administered weekly for the first three months, then biweekly for the next three months, and then monthly.

[0079] combination The present disclosure provides an anti-CD19 antibody for treating a patient with a blood cancer, the patient having a peripheral NK cell count of less than 100 cells / μl at baseline, wherein the anti-CD19 antibody is administered in combination with one or more drugs. In one embodiment of the present disclosure, the anti-CD19 antibody is administered in combination with a drug. In another embodiment of the present disclosure, the anti-CD19 antibody is administered in combination with one or more additional drugs or an additional drug. In one aspect, the drug is an additional drug.

[0080] In one embodiment of the present disclosure, the drug is a biological drug or a chemotherapeutic agent.In another embodiment of the present disclosure, the drug is a therapeutic antibody or antibody fragment, nitrogen mustard, purine analog, thalidomide analog, phosphoinositide 3-kinase inhibitor, BCL-2 inhibitor or Bruton's tyrosine kinase (BTK) inhibitor.In another embodiment, the drug is rituximab, R-CHOP, cyclophosphamide, chlorambucil, uramustine, ifosfamide, melphalan, bendamustine, mercaptopurine, azathioprine, thioguanine, fludarabine, thalidomide, lenalidomide, pomalidomide, idelalisib, duvelisib, copanlisib, ibrutinib or venetoclax.

[0081] In another embodiment, the present disclosure provides an anti-CD19 antibody for use in treating a patient with a hematological cancer, the patient having a baseline peripheral NK cell count of 100 cells / μl or less, and the anti-CD19 antibody is administered in combination with rituximab, R-CHOP, cyclophosphamide, chlorambucil, uramustine, ifosfamide, melphalan, bendamustine, mercaptopurine, azathioprine, thioguanine, fludarabine, thalidomide, lenalidomide, pomalidomide, idelalisib, duvelisib, copanlisib, ibrutinib, or venetoclax. In a further embodiment, the present disclosure provides an anti-CD19 antibody for treatment of a patient with a hematological cancer, the patient having a baseline peripheral NK cell count of 100 cells / μl or less, and the anti-CD19 antibody is administered in combination with bendamustine.

[0082] Indications and patients The present disclosure provides an anti-CD19 antibody for the treatment of a patient with a hematological cancer, the patient having a peripheral NK cell count of 100 cells / μl or less at baseline, the patient having chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), small lymphocytic lymphoma (SLL), or acute lymphoblastic leukemia (ALL). In another embodiment, the patient has a non-Hodgkin's lymphoma. In a further embodiment, the non-Hodgkin's lymphoma is selected from the group consisting of follicular lymphoma, small lymphocytic lymphoma, mucosa-associated lymphoid tissue, marginal zone lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, and mantle cell lymphoma. In a further embodiment, the non-Hodgkin's lymphoma is relapsed or refractory diffuse large B-cell lymphoma (rr DLBCL). In another embodiment, the hematological cancer patient has diffuse large B-cell lymphoma and is ineligible for high-dose chemotherapy (HDC) and / or autologous stem cell transplant (ASCT). In another embodiment, the hematological cancer patient has relapsed or refractory diffuse large B-cell lymphoma (rr DLBCL) and is ineligible for high-dose chemotherapy (HDC) and / or autologous stem cell transplant (ASCT).

[0083] In another embodiment, the hematological cancer patient has diffuse large B-cell lymphoma, and the patient is selected based on one or more of the following criteria: 1. Age 18 or older 2. Histologically confirmed diagnosis of disease that has transformed from a previous diagnosis of DLBCL NOS, THRLBCL, EBV-positive DLBCL, composite lymphoma with a DLBCL component with DLBCL relapse after DLBCL treatment, or low-grade lymphoma (i.e., indolent pathologies such as follicular lymphoma, marginal zone lymphoma, etc.) to DLBCL with DLBCL relapse after DLBCL treatment, according to the World Health Organization (WHO, 2008) classification. 3. Fresh tumor tissue for central pathology review must be provided as an adjunct to participation in this study. If fresh tumor tissue samples cannot be obtained, archival paraffin-embedded tumor tissue obtained within 3 years prior to screening for this protocol must be available for this purpose. 4. The patient must have: 1. Relapsed or refractory DLBCL 2. At least one bidimensionally measurable disease site. The lesion must have a maximum transverse diameter of ≥ 1.5 cm and a maximum perpendicular diameter of ≥ 1.0 cm at baseline. The lesion must be positive on PET scan. 3. Received at least one, but no more than three, prior lines of systemic therapy for the treatment of DLBCL. At least one prior line of therapy must have included a CD20 target. 4. ECOG 0-2 5. Patients after ASCT failure or considered in the investigator's opinion to be currently ineligible for HDC with subsequent ASCT. 6. Patients must meet the following clinical laboratory criteria at screening: a) ANC ≥ 1.5 × 109 / L (except following bone marrow involvement by DLBCL) b) PLT ≥ 90 x 109 / L (except following bone marrow involvement by DLBCL) and absence of active bleeding c) Total serum bilirubin ≤ 2.5 x ULN, except following Gilbert's syndrome (or a pattern consistent with Gilbert's syndrome) or with documented liver involvement by lymphoma. Patients with Gilbert's syndrome or with documented liver involvement by lymphoma may be included if their total bilirubin is ≤ 5 x ULN. d) If liver involvement by lymphoma is documented, ALT, AST, and AP ≤ 3 x ULN or < 5 x ULN e) Serum creatinine ≤ 2.0 x ULN or creatinine clearance ≥ 40 mL / min calculated using the standard Cockcroft-Gault formula (Cockroft & Gault, 1976). 7. Women of childbearing potential (FCBP) must have a negative pregnancy test prior to enrollment. FCBP must undertake to use highly effective contraception without interruption during the study and for 3, 6, or 12 months after the last dose of MOR00208, BEN, or RTX, whichever occurs later. FCBP must refrain from breast-feeding and from donating blood or oocytes during the course of the study and for 3, 6, or 12 months after the last dose of MOR00208, BEN, or RTX, whichever occurs later. Restrictions on blood donation apply equally to women who are not of childbearing potential. 8. Men must use effective contraception without interruption during study participation and for 3, 6, or 12 months after the last dose of MOR00208, BEN, or RTX, whichever is later, if the patient is sexually active with FCBP. Men must abstain from donating blood or sperm during study participation and for 3, 6, or 12 months after the last dose of MOR00208, BEN, or RTX, whichever is later. 9. In the opinion of the investigator, the patient: a) be able to comply with all study-related procedures, drug use and evaluations b) be able to understand and give informed consent c) Must not be perceived as potentially unreliable and / or uncooperative.

[0084] In another embodiment, the hematological cancer patient has diffuse large B-cell lymphoma, and the patient is excluded based on one or more of the following criteria: 1. Patients with any other histological type of lymphoma, including, for example, primary mediastinal (thymic) large B-cell lymphoma (PMBL) or Burkitt's lymphoma, primary refractory DLBCL, patients with known "double / triple hit" DLBCL genes, and CNS lymphoma involvement in current or past medical history 2. Patients who underwent major surgery within 30 days prior to the first day of administration 3. Patients with the following within 14 days prior to dosing on Day 1: a) No discontinuation of CD20-targeted therapy, chemotherapy, radiation therapy, investigational anticancer therapy, or other lymphoma-specific therapy b) received a live vaccine c) Parenteral antimicrobial therapy as needed for active intercurrent systemic infections 4. Patients with: a) In the opinion of the investigator, has not adequately recovered from the adverse toxic effects of previous therapy, major surgery, or significant trauma b) Previously treated with CD19-targeted therapy or BEN c) History of a previous severe allergic reaction to MOR00208, RTX, mouse protein, or BEN, or compounds with similar biological or chemical composition to the excipients contained in the investigational drug formulation d) Have undergone ASCT within 3 months prior to signing the informed consent form. Patients with a more distant history of ASCT must demonstrate complete hematologic recovery before enrolling in the study. e) Previous allogeneic stem cell transplant f) Concurrent use of other anti-cancer drugs or experimental treatments 5. History of malignancy other than DLBCL, provided that the patient has been disease-free for at least 3 years prior to screening. Exceptions to the 3+ year limit include a history of: a) Basal cell carcinoma of the skin b) Squamous cell carcinoma of the skin c) Carcinoma in situ of the cervix, breast and bladder d) incidental histological findings of prostate cancer (tumor / node / metastasis [TNM] stage of T1a or T1b); 6. Patients with: a) Positive hepatitis B and / or C serology b) known seropositivity or history of active HIV viral infection c) Evidence of active, severe, uncontrolled systemic infection or sepsis d) History or evidence of a severely immunocompromised state e) History or evidence of severe liver damage (total serum bilirubin >3 mg / dL), jaundice secondary to Gilbert's syndrome, or documented liver involvement by lymphoma f) History or evidence of clinically significant cardiovascular, cerebrovascular, CNS, and / or other disease that, in the investigator's opinion, would prevent participation in the study or impair the patient's ability to give informed consent.

[0085] Treatment method The present disclosure provides methods of treating a patient with a hematological cancer by administration of an anti-CD19 antibody, wherein the patient has a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less.

[0086] The present disclosure provides methods of treating a patient with a hematological cancer by administration of an anti-CD19 antibody, wherein the patient has a baseline peripheral NK cell count of less than 100 cells / μl, less than 90 cells / μl, less than 80 cells / μl, less than 70 cells / μl, less than 60 cells / μl, or less than 50 cells / μl.

[0087] In another embodiment, the present disclosure refers to a pharmaceutical composition comprising an anti-CD19 antibody disclosed herein for use in treating a hematological cancer. In another embodiment, the present disclosure refers to the use of the pharmaceutical composition comprising an anti-CD19 antibody disclosed herein in the preparation of a medicament for treating a hematological cancer. In another embodiment, the present disclosure refers to the use of the pharmaceutical composition comprising an anti-CD19 antibody disclosed herein for treating a hematological cancer. In another embodiment, the hematological cancer is chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), small lymphocytic lymphoma (SLL), or acute lymphoblastic leukemia (ALL). In another embodiment, the hematological cancer is non-Hodgkin's lymphoma. In a further embodiment, the non-Hodgkin's lymphoma is selected from the group consisting of follicular lymphoma, small lymphocytic lymphoma, mucosa-associated lymphoid tissue, marginal zone lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, and mantle cell lymphoma. In a further embodiment, the non-Hodgkin's lymphoma is relapsed or refractory diffuse large B-cell lymphoma (rr DLBCL). In another embodiment, the hematological cancer patient has diffuse large B-cell lymphoma and is ineligible for high-dose chemotherapy (HDC) and / or autologous stem cell transplant (ASCT).

[0088] In another aspect, provided herein are methods for treating hematological cancer in a patient, the patient having a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less, the method comprising administering a therapeutically effective amount of a pharmaceutical composition comprising an anti-CD19 antibody disclosed herein. In one embodiment, the patient is resistant, non-responsive, or inadequately responsive to treatment with no more than one to three prior lines of therapy, including one anti-CD20 targeted therapy (e.g., the antibody rituximab). In a further embodiment, the patient is not a candidate for high-dose chemotherapy and autologous stem cell transplantation. In a preferred embodiment, the patient is human. In an alternative aspect, the patient is a rodent, such as a rat or mouse. In another embodiment, the patient is suffering from a hematological cancer, such as non-Hodgkin's lymphoma. In further embodiments, the non-Hodgkin's lymphoma is selected from the group consisting of follicular lymphoma, small lymphocytic lymphoma, mucosa-associated lymphoid tissue, marginal zone lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, and mantle cell lymphoma. In further embodiments, the non-Hodgkin's lymphoma is relapsed or refractory diffuse large B-cell lymphoma (rr DLBCL).

[0089] In another aspect, the disclosure provides for the use of an anti-CD19 antibody in the manufacture of a medicament for use in treating a patient with a hematological cancer, wherein the patient has a baseline peripheral NK cell count of 100 cells / μl or less, 90 cells / μl or less, 80 cells / μl or less, 70 cells / μl or less, 60 cells / μl or less, or 50 cells / μl or less.

[0090] In another aspect, the present disclosure provides the use of an anti-CD19 antibody in the manufacture of a medicament for use in treating a hematological cancer. In another embodiment, the present disclosure refers to the use of the anti-CD19 antibody disclosed herein in the preparation of a medicament for treating a hematological cancer. In another embodiment, the present disclosure refers to the use of the pharmaceutical composition comprising the anti-CD19 antibody disclosed herein for the treatment of a hematological cancer. In another embodiment, the hematological cancer is chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), small lymphocytic lymphoma (SLL), or acute lymphoblastic leukemia (ALL). In another embodiment, the hematological cancer is non-Hodgkin's lymphoma. In a further embodiment, the non-Hodgkin's lymphoma is selected from the group consisting of follicular lymphoma, small lymphocytic lymphoma, mucosa-associated lymphoid tissue, marginal zone lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, and mantle cell lymphoma. In a further embodiment, the non-Hodgkin's lymphoma is relapsed or refractory diffuse large B-cell lymphoma (rr DLBCL). In another embodiment, the hematological cancer patient has diffuse large B-cell lymphoma and is ineligible for high-dose chemotherapy (HDC) and / or autologous stem cell transplant (ASCT).

[0091] In some embodiments, the anti-CD19 antibodies disclosed herein are administered intravenously. In other aspects, the anti-CD19 antibodies disclosed herein are administered subcutaneously, intraarticularly, or intraspinally.

[0092] method The present disclosure provides a method for selecting a patient with hematological cancer expected to benefit from therapeutic administration of an anti-CD19 antibody, the method comprising the steps of: a) providing a blood sample obtained from said patient prior to treatment with said anti-CD19 antibody; b) determining peripheral NK cell numbers, and c) Patients are selected on the basis that they have a peripheral NK cell count of 100 cells / μl or less at baseline.

[0093] In an embodiment of the present disclosure, the method further comprises the steps of: d) Treatment of selected patients with anti-CD19 antibodies.

[0094] The present disclosure provides a method for selecting a patient with hematological cancer who is expected to benefit from therapeutic administration of an anti-CD19 antibody in combination with a drug, the method comprising the steps of: a) providing a blood sample obtained from said patient prior to treatment with said anti-CD19 antibody; b) determining peripheral NK cell numbers, and c) Patients are selected on the basis that they have a peripheral NK cell count of 100 cells / μl or less at baseline.

[0095] In an embodiment of the present disclosure, the method further comprises the steps of: d) Treatment of selected patients with anti-CD19 antibodies.

[0096] The present disclosure provides a method for selecting a patient with hematological cancer predicted to benefit from therapeutic administration of an anti-CD19 antibody, the method comprising the steps of: a) providing a blood sample obtained from said patient prior to treatment with said anti-CD19 antibody; b) determining peripheral NK cell numbers, and c) Patients are selected on the basis that they have a peripheral NK cell count of 100 cells / μl or less at baseline.

[0097] In an embodiment of the present disclosure, the method further comprises the steps of: d) Treatment of selected patients with anti-CD19 antibodies.

[0098] The present disclosure provides a method for selecting a patient with hematological cancer predicted to benefit from therapeutic administration of an anti-CD19 antibody in combination with a drug, the method comprising the steps of: a) providing a blood sample obtained from said patient prior to treatment with said anti-CD19 antibody; b) determining peripheral NK cell numbers, and c) Patients are selected on the basis that they have a peripheral NK cell count of 100 cells / μl or less at baseline.

[0099] In an embodiment of the present disclosure, the method further comprises the steps of: d) Treatment of selected patients with anti-CD19 antibodies.

[0100] In one embodiment of the present disclosure, the drug administered in combination with the anti-CD19 antibody is a biological drug or a chemotherapeutic agent.In another embodiment of the present disclosure, the drug is a therapeutic antibody or antibody fragment, nitrogen mustard, a purine analog, a thalidomide analog, a phosphoinositide 3-kinase inhibitor, a BCL-2 inhibitor or a Bruton's tyrosine kinase (BTK) inhibitor.In a further embodiment, the drug is rituximab, R-CHOP, cyclophosphamide, chlorambucil, uramustine, ifosfamide, melphalan, bendamustine, mercaptopurine, azathioprine, thioguanine, fludarabine, thalidomide, lenalidomide, pomalidomide, idelalisib, duvelisib, copanlisib, ibrutinib or venetoclax.

[0101] The present disclosure provides a method for identifying a patient with hematological cancer predicted to benefit from therapeutic administration of an anti-CD19 antibody, the method comprising the steps of: a) providing a blood sample obtained from said patient prior to treatment with said anti-CD19 antibody; b) determining peripheral NK cell numbers, and c) Patients are selected on the basis that they have a peripheral NK cell count of 100 cells / μl or less at baseline.

[0102] In an embodiment of the present disclosure, the method further comprises the steps of: d) Treatment of selected patients with anti-CD19 antibodies.

[0103] The present disclosure provides a method of treating hematological cancer by administering an anti-CD19 antibody to a hematological cancer patient, wherein the patient has been selected according to a method comprising the steps of: a) providing a blood sample obtained from said patient prior to treatment with said anti-CD19 antibody; b) determining peripheral NK cell numbers, and c) Patients are selected on the basis that they have a peripheral NK cell count of 100 cells / μl or less at baseline.

[0104] In an embodiment of the present disclosure, the method further comprises the steps of: d) Treatment of selected patients with anti-CD19 antibodies.

[0105] The present disclosure provides a method for selecting a patient with hematological cancer predicted to benefit from therapeutic administration of an anti-CD19 antibody, the method comprising the steps of: a) providing a blood sample obtained from said patient prior to treatment with said anti-CD19 antibody; b) determining peripheral NK cell numbers, and c) Patients are selected on the basis that they have a peripheral NK cell count of 100 cells / μl or less at baseline.

[0106] In an embodiment of the present disclosure, the method further comprises the steps of: d) Treatment of selected patients with anti-CD19 antibodies.

[0107] In another embodiment of the present disclosure, the expected benefit from therapeutic administration of an anti-CD19 antibody is improved progression-free survival (PFS), improved objective response rate (ORR), improved duration of response (DoR), improved overall survival (OS), or improved time to progression (TTP), or a combination thereof.

[0108] In another embodiment of the present disclosure, the expected benefit from the therapeutic administration of an anti-CD19 antibody is improved progression-free survival (PFS) compared to administration of an anti-CD20 antibody, improved objective response rate (ORR) compared to administration of an anti-CD20 antibody, improved duration of response (DoR) compared to administration of an anti-CD20 antibody, improved overall survival (OS) compared to administration of an anti-CD20 antibody, or improved time to progression (TTP) compared to administration of an anti-CD20 antibody, or a combination thereof.

[0109] In another embodiment of the present disclosure, the expected benefit from therapeutic administration of an anti-CD19 antibody is: (i) progression-free survival (PFS) of at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 12 months, at least 13 months, at least 14 months, at least 15 months, at least 16 months, at least 17 months, at least 18 months, at least 19 months, or at least 20 months; (ii) an objective response rate (ORR) of at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 80%; (iii) duration of response (DoR) of at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, at least 48 months, or at least 54 months; (iv) overall survival (OS) of at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, at least 24 months, at least 30 months, at least 36 months, at least 42 months, at least 48 months, or at least 54 months; or (v) A combination of one or more of the above. In another embodiment of the disclosure, the anti-CD19 antibody is administered in combination with an agent disclosed herein.

[0110] In another embodiment of the present disclosure, the predicted benefit from therapeutic administration of an anti-CD19 antibody is improved progression-free survival (PFS) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent, improved objective response rate (ORR) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent, improved duration of response (DoR) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent, improved overall survival (OS) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent, or improved time to progression (TTP) compared to administration of an anti-CD20 antibody and a chemotherapeutic agent. In a further embodiment, the anti-CD20 antibody is rituximab or a biosimilar thereof. In a further embodiment, the chemotherapeutic agent comprises one or more of cyclophosphamide, adriamycin, vincristine, or prednisone.

[0111] In another embodiment of the present disclosure, the expected benefit from the therapeutic administration of an anti-CD19 antibody is improved progression-free survival (PFS) compared to administration of R-CHOP, improved objective response rate (ORR) compared to administration of R-CHOP, improved duration of response (DoR) compared to administration of R-CHOP, improved overall survival (OS) compared to administration of R-CHOP, or improved time to progression (TTP) compared to administration of R-CHOP.

[0112] In another embodiment of the disclosure, the predicted benefit from therapeutic administration of an anti-CD19 antibody is an increase in one or more of the following characteristics: (i) progression-free survival (PFS), (ii) objective response rate (ORR); (iii) duration of response (DoR); (iv) overall survival (OS); (v) Time to progression (TTP).

[0113] In another embodiment, the increase in one or more of features (i)-(v) is compared to a treatment comprising an anti-CD20 antibody. In a further embodiment, the increase in one or more of features (i)-(v) is compared to a treatment comprising an anti-CD20 antibody and a chemotherapeutic agent. In a further embodiment, the anti-CD20 antibody is rituximab or a biosimilar thereof. In a further embodiment, the increase in one or more of features (i)-(v) is compared to a treatment comprising an anti-CD20 antibody and one or more of cyclophosphamide, adriamycin, vincristine, or prednisone. In a further embodiment, the increase in one or more of features (i)-(v) is compared to a treatment comprising R-CHOP.

[0114] In an embodiment of the present disclosure, in the method of selecting a hematological cancer patient predicted to benefit from therapeutic administration of an anti-CD19 antibody, the hematological cancer patient has chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), small lymphocytic lymphoma (SLL), or acute lymphoblastic leukemia (ALL). In a further embodiment, the hematological cancer patient has non-Hodgkin's lymphoma. In a further embodiment, the hematological cancer patient has non-Hodgkin's lymphoma, wherein the non-Hodgkin's lymphoma is selected from the group consisting of follicular lymphoma, small lymphocytic lymphoma, mucosa-associated lymphoid tissue, marginal zone lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, and mantle cell lymphoma. In a further embodiment, the hematological cancer patient has relapsed or refractory diffuse large B-cell lymphoma (rr DLBCL).

[0115] In a further embodiment of the disclosure, the anti-CD19 antibody of the method for selecting a hematological cancer patient predicted to benefit from therapeutic administration of an anti-CD19 antibody comprises an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6). array The variable heavy chain of EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7) and array The variable light chain of the compound of formula (SEQ ID NO: 8) is: DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK.

[0116] In a further embodiment, the anti-CD19 antibody has the sequence EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP A heavy chain having CPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11) and array and a light chain having the sequence DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12).

[0117] In another embodiment, an anti-CD19 antibody for the treatment of a patient with hematological cancer comprises a variable heavy chain and a variable light chain having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to the variable heavy chain of SEQ ID NO:7 and the variable light chain of SEQ ID NO:8.

[0118] In embodiments, an anti-CD19 antibody for the treatment of a patient with hematological cancer comprises a variable heavy chain and a variable light chain having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the variable heavy chain of SEQ ID NO: 7 and the variable light chain of SEQ ID NO: 8, wherein the anti-CD19 antibody comprises an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6). In another embodiment, the heavy chain region of the anti-CD19 antibody comprises amino acids 239D and 332E, and Fc numbering is according to the EU index as in Kabat.

[0119] In further embodiments, the anti-CD19 antibody for the treatment of patients with hematological cancer comprises a heavy chain and a light chain having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to the heavy chain of SEQ ID NO:7 and the light chain of SEQ ID NO:8.

[0120] In a further embodiment, an anti-CD19 antibody for the treatment of a patient with hematological cancer comprises a heavy chain and a light chain having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the heavy chain of SEQ ID NO: 7 and the light chain of SEQ ID NO: 8, wherein the anti-CD19 antibody comprises an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6). In another embodiment, the heavy chain region of the anti-CD19 antibody comprises amino acids 239D and 332E, and Fc numbering is according to the EU index as in Kabat.

[0121] In a further embodiment, the present disclosure provides a kit comprising a means for determining peripheral NK cell numbers in a hematological cancer patient treated with an anti-CD19 antibody.

[0122] In a further embodiment, the present disclosure refers to the use of peripheral NK cell count as a biomarker for predicting the sensitivity of a hematological cancer patient to treatment with an anti-CD19 antibody. In another embodiment, the baseline peripheral NK cell count is 100 cells / μL or less, 90 cells / μL or less, 80 cells / μL or less, 70 cells / μL or less, 60 cells / μL or less, or 50 cells / μL or less. In other embodiments, the baseline peripheral NK cell count is 1 to a maximum of 100 cells / μL, 10 to a maximum of 100 cells / μL, 20 to a maximum of 100 cells / μL, 30 to a maximum of 100 cells / μL, 40 to a maximum of 100 cells / μL, 50 to a maximum of 100 cells / μL, 60 to a maximum of 100 cells / μL, 70 to a maximum of 100 cells / μL, or 80 to a maximum of 100 cells / μL.

[0123] In a further embodiment, the present disclosure refers to the use of peripheral NK cell count as a biomarker for predicting the sensitivity of a hematological cancer patient to treatment with an anti-CD19 antibody. In another embodiment, the baseline peripheral NK cell count is less than 100 cells / μl, less than 90 cells / μl, less than 80 cells / μl, less than 70 cells / μl, less than 60 cells / μl, or less than 50 cells / μl. In other embodiments, the baseline peripheral NK cell count is between 1 and 100 cells / μl, between 10 and 100 cells / μl, between 20 and 100 cells / μl, between 30 and 100 cells / μl, between 40 and 100 cells / μl, between 50 and 100 cells / μl, between 60 and 100 cells / μl, between 70 and 100 cells / μl, or between 80 and 100 cells / μl.

[0124] Antibody sequence [Table 1] TIFF2026004466000009.tif140161 TIFF2026004466000010.tif201160

[0125] Example ADCC activity of MOR00208 and rituximab in DLBCL, MCL, and CLL cell lines at different E:T ratios

[0126] Example 1: Characterization of CD19 and CD20 expression on tested cell lines Methods and data analysis In this study, we used the QuantiBRITE™ system to quantify the amount of bound phycoerythrin (PE)-labeled CD19 and CD20 antibodies per cell according to the manufacturer's instructions. The QuantiBRITE™ system is based on four sets of beads coated with different pre-calibrated levels of PE molecules, which were used to correlate mean fluorescence intensity (MFI) values ​​with the number of PE molecules per bead. For each individual cell type, the MFI measured upon staining with the PE-labeled antibodies was applied to a linear regression equation to calculate the respective bound antibody (ABC) values ​​per cell. The ABC values ​​directly correlate with the number of CD19 and CD20 molecules per cell because Biolegend CD19-PE (Biolegend #302208, clone HIB19) and CD20-PE (Biolegend #302306, clone 2H7) antibodies carry only one PE molecule per antibody. At a 1:1 labeling ratio of fluorescent dye / protein (F / P), the MESF (molecules of equivalent soluble fluorescent dye) value corresponds to the ABC value according to the formula: MESF / ABC = effective F / P. GraphPad PRISM™ software was used to convert MFI to ABC values.

[0127] result CD19 and CD20 expression levels were analyzed in Toledo (DLBCL), MEC-1 (CLL), and JVM-2 (MCL) cells. The QuantiBRITE™ system combined with PE-labeled anti-CD19 and anti-CD20 antibodies was used to determine the CD19 and CD20 expression levels in the tested B-cell tumor cell lines. For the DLBCL cell line Toledo, CD19 and CD20 expression levels of 35,721 and 28,008 bound antibodies (ABC) per cell were determined (Table 1). CD19 and CD20 expression on MEC-1 cells (CLL) were quantified as 60,925 and 71,320 ABC, respectively, while JVM-2 cells (MCL) showed a CD19 expression level of 26,157 and a CD20 expression level of 15,540 ABC. [Table 2]

[0128] Example 2: ADCC activity assay at varying E:T ratios Methods and data analysis Antibody-dependent cell-mediated cytotoxicity (ADCC) is an immune cytotoxic effector mechanism that primarily relies on the interaction of antibodies with Fc receptors on NK cells. ADCC is triggered when an antibody binds to a specific antigen on the surface of a target cell, such as CD19 or CD20 on a cancer cell, and the Fc fragment of the antibody interacts with an Fc receptor on an effector cell, such as an NK cell. This interaction activates the effector cell, and lysis of the target cell is induced by the release of perforin and granzymes.

[0129] For effector cell preparation, peripheral blood mononuclear cells (PBMCs) were isolated from whole blood of healthy volunteers by density gradient centrifugation using Biocoll separator and SepMate tubes. NK cells were then isolated from PBMCs using a MACS kit according to the manufacturer's protocol. One CLL cell line (MEC-1), one MCL cell line (JVM-2), and one DLBCL cell line (Toledo) were stained with 1 μM carboxyfluorescein succinimidyl ester (CFSE) for 3 minutes at room temperature before incubation with antibodies and NK effector cells.

[0130] For ADCC experiments, 2 x 10 cells per well 4 DLBCL, MCL, or CLL target cells were incubated with NK cells as effector cells at various effector-to-target (E:T) ratios, and MOR00208 or rituximab was incubated at a concentration of 10 μg / ml for 2 hours at 37°C and 5% CO. Nonspecific NK cell-mediated killing of tumor cells was determined by incubation of NK cells with target cells in the absence of antibody.

[0131] A flow cytometry-based assay was used to measure target cell killing by quantifying dead and live cells using the DNA intercalating dye 4',6-diamidino-2-phenylindole (DAPI), which is membrane-impermeable and intercalates only into the DNA of dead cells with damaged membranes but is excluded from live cells with intact membranes. Cells were stained with DAPI at a final concentration of 1 μg / ml and incubated on ice for 10 min before FACS measurement.

[0132] Raw data were collected on a FACS Verse instrument and analyzed with FlowJo software. Cell populations were gated for live (DAPI-negative) and dead target cells (DAPI-positive). Data were exported to Microsoft Excel®, and the percentage of dead cells and specific death were calculated using the following formulas: % Dead Cells = Dead Target Cells / (Dead Target Cells + Live Target Cells) x 100 (Sample) % specific killing = % dead cells - % dead cells in NK and target cell control (no antibody).

[0133] Data analysis was performed using the statistical software package R and R Studio (version 1.0.153 RStudio, Inc.). For each independent experiment, the percent specific killing measured in triplicate was summarized by geometric mean and its standard deviation. Furthermore, the specific killing ratio was calculated from the percent specific killing value by normalizing to the median value of rituximab. For visualization, the geometric mean and its 95% confidence interval were calculated. The confidence interval of the geometric mean was calculated by bootstrap resampling using 1,000 replicates.

[0134] To summarize data from all experiments per cell line, the specific killing ratios for each donor were combined in a two-step process. First, triplicate values ​​from individual experiments were aggregated into a geometric mean. Subsequently, the geometric means from independent experiments were combined into a single geometric mean for each E:T ratio for each individual donor. For each E:T ratio, the median and its confidence interval were calculated by bootstrap resampling with 10,000 replicates based on the summarized geometric means for each individual donor (R Core Team 2017, Davison and Hinkley 1997, Wickham 2017).

[0135] result The ADCC activity of the Fc-enhanced anti-CD19 antibody MOR00208 and the anti-CD20 antibody rituximab against Toledo (DLBCL), MEC-1 (CLL), and JVM-2 (MCL) cells was measured after a 2-hour incubation with NK cells isolated from healthy human donors.

[0136] The antitumor activity of MOR00208 and rituximab was evaluated at an antibody concentration of 10 μg / ml at E:T ratios of 0.1:1, 0.3:1, 1:1, 3:1, and 6:1. E:T ratios ranging from 0.1:1 to 6:1 were selected as the lower and upper limits of the assay. The lowest ratio (1 NK cell to 10 tumor cells, ratio 0.1:1) was determined by the minimal detectable ADCC signal in such an in vitro assay. The upper limit (6 NK cells to 1 tumor cell, 6:1) was selected as the ratio at which maximal lysis was achieved under the conditions of such an in vitro assay.

[0137] Figure 1 shows representative results from individual experiments for each target cell line, presented as % specific killing, and Figure 2 shows the specific killing ratio of MOR00208 normalized to rituximab.

[0138] Figure 1 shows the results of one representative assay of specific cell killing in MEC-1 cells mediated by MOR00208 (black) or rituximab (white) in a 2-hour assay at 37°C in the presence of NK cells. At E:T ratios of 3:1 and 6:1, mean specific killing levels of 40-65% mediated by MOR00208 and 34-60% for rituximab were found in MEC-1 cells. In an exemplary JVM-2 ADCC assay, specific killing of MOR00208 versus rituximab also increased at E:T ratios of 3:1 and 6:1, ranging from 46-56% versus 39-48%. Toledo specific cell killing was similar at the two higher E:T ratios for MOR00208 and rituximab, with values ​​of approximately 60%. At a low E:T ratio of 0.1:1, specific killing of rituximab was 3% in MEC-1 cells, while MOR00208 showed 6% specific killing. Similar findings were seen in JVM-2 cells, where specific cell killing was increased with MOR00208 (3.4%) vs. rituximab (0.5%), and in Toledo cells, where cell killing was increased with MOR00208 (8%) vs. rituximab (3%), at an E:T ratio of 0.1:1.

[0139] The median ratios of MOR00208 (black triangles) and rituximab (circles) specific killing to rituximab increased 1.9-6.9-fold at an E:T ratio of 0.1:1 in all representative individual ADCC experiments using DLBCL, MCL, and CLL cell lines (Figure 2). At higher E:T ratios of 3:1 and 6:1, specific killing reached saturation levels (see Figure 1), resulting in only minor differences in the specific killing ratios of MOR00208 to rituximab (Figure 2). All experimental data points were evaluated in triplicate. In summary, E:T titrations were performed in ADCC assays using three B-cell tumor cell lines and NK cells from 61 blood samples isolated from 33 healthy blood donors. ADCC activity of MOR00208 and rituximab was evaluated in eight donors for JVM-2 cells and 10 donors for Toledo cells, with two independent experiments for each donor. MEC-1 cells were tested in eight donors in two independent experiments and nine additional donors in a single experiment.

[0140] Figure 3 shows the specific killing ratios normalized to rituximab for all experiments performed with each cell line. Here, each circle or triangle represents the geometric mean of two independent experiments performed in triplicate using NK cells from one individual blood donor. A 5.3- or 2.5-fold specific killing ratio of MOR00208 normalized to rituximab was found in JVM-2 or Toledo cells at an E:T ratio of 0.1:1, shown as the median value across multiple donors (Figure 3). NK cells from individual donors showed up to a 20- or 30-fold increase in the specific killing ratio of MOR00208 compared to rituximab at the lowest E:T ratios (e.g., donor 296 with JVM-2 target cells or donor 299 with Toledo target cells) (data not shown). This effect was weaker in MEC-1 cells, where multiple ADCC assays using NK cells isolated from 17 different donors resulted in an average 1.6-fold increase in the specific killing ratio of MOR00208 compared to rituximab at an E:T ratio of 0.1:1. At higher E:T ratios, the increase in the specific killing ratio of MOR00208 compared to rituximab was less pronounced in all B-cell tumor cell lines tested, whereas at lower E:T ratios, MOR00208 was clearly superior to rituximab. It should be noted that the confidence intervals for MOR00208, as shown in Figure 3, do not overlap with those for rituximab, except for Toledo cells at an E:T ratio of 6:1, suggesting a general robustness regarding the observed superiority of MOR00208 versus rituximab. In conclusion, the specific killing ratio increased towards lower E:T ratios and was most pronounced for JVM-2 and Toledo cells, although a consistent effect was seen for MEC-1 cells.

[0141] The monoclonal antibody MOR00208 targets the CD19 antigen on B cells and contains two mutations (S239D and I332E) in the Fc region, enhancing antibody-dependent cell-mediated cytotoxicity (ADCC). ADCC is an important mechanism of cancer cell killing, primarily mediated by tumor-infiltrating NK cells. Bhat and Watzl (2007) demonstrated increased sequential killing of NK cells in the presence of rituximab, with maximal efficacy observed at low E:T ratios of 0.05:1, 0.1:1, and 0.2:1. Furthermore, Fc enhancement of antibodies has been reported to result in increased sequential killing of NK cells compared to non-enhanced versions of CD33-specific antibodies (Romain et al. 2014). Here, across a wide range of healthy donors, using one cell line derived from DLBCL, one derived from MCL, and one derived from CLL mediated by NK cells, we demonstrate a 1.6- to 5.3-fold increase in specific killing of MOR00208 normalized to rituximab at an E:T ratio as low as 0.1:1. The specific killing ratio increased toward lower E:T ratios, most pronounced for JVM-2 and Toledo, but a consistent effect was observed for MEC-1 cells. At higher E:T ratios of 3:1 and 6:1, MOR00208-mediated specific killing was similar to rituximab at a saturating antibody concentration of 10 μg / ml.

[0142] The increased specific killing of MOR00208 compared to rituximab at low E:T ratios was confirmed in freshly isolated NK cells from 33 healthy donors in 61 independent experiments in Toledo, JVM-2, and MEC-1 cells. These results demonstrate increased serial killing of NK cells at lower E:T ratios and provide evidence that MOR00208 has increased antitumor activity in DLBCL, MCL, and CLL patients with low NK cell numbers. In conclusion, the ADCC activity of MOR00208 showed its most significant advantage over rituximab under conditions in which NK cells are limited. Therefore, therapy including the use of MOR00208 is preferable to standard treatment (e.g., rituximab) for patients with low baseline NKCC.

[0143] Example 3: T cell and NK cell counting As an example, peripheral T and NK cell counts can be performed according to the following procedure. T cells are a type of lymphocyte (a subtype of white blood cell) that plays a central role in cell-mediated immunity. They can be distinguished from other lymphocytes, such as B cells and NK cells, by the presence of T cell receptors on their cell surface.

[0144] Natural killer cells, or NK cells, are a type of cytotoxic lymphocyte essential to the innate immune system. NK cells provide a rapid response to virus-infected cells, acting approximately three days after infection and responding to tumor formation. Typically, immune cells detect major histocompatibility complexes (MHC) displayed on the surface of infected cells, triggering cytokine release and causing lysis or apoptosis. However, NK cells are unique because they have the ability to recognize stressed cells in the absence of antibodies and MHC, enabling a much faster immune response.

[0145] Materials and Methods TriTest CD3 FITC / CD16+CD56 PE / CD45 PerCP (including TruCOUNT tubes), BD Biosciences, Catalog Number 340403 (USA), 342442 (Europe). Pipettors and pipette tips capable of delivering 20 μL, 50 μL, and 450 μL, Gilson Inc. FACS Lysing Solutions, BD Biosciences, Catalog Number 349202. Instrument: Flow cytometer, Vortex

[0146] Flow Cytometry Background: Whole blood is stained with fluorochrome-conjugated antibodies (TriTEST) that specifically bind to leukocyte surface antigens. Cells pass through a laser beam, scattering the laser light. Stained cells fluoresce. These scattering and fluorescence signals, detected by the instrument, provide information about cell size, internal complexity, and relative fluorescence intensity. TriTEST reagents use a fluorescent trigger, allowing direct fluorescent gating of NK cell and T cell lymphocyte populations, reducing contamination of unlysed or nucleated red blood cells within the gate.

[0147] staining: For each patient sample, a TruCOUNT tube is labeled with the sample identification number. 20 μL of TriTEST CD3 / CD16+CD56 / CD45 Reagent is pipetted into the bottom of the tube. 50 μL of well-mixed anticoagulated whole blood is pipetted into the bottom of the tube. Anticoagulated blood (EDTA) stored at room temperature (20-25°C) must be stained within 24 hours of extraction and analyzed within 6 hours of staining (store at room temperature and protect from light). Gently vortex the tube to mix. Incubate the tube in the dark at room temperature (20-25°C) for 15 minutes. Add 450 μL 1x FACS Lysing Solution to the tube. Vortex the tube and incubate again in the dark at room temperature (20-25°C) for 15 minutes.

[0148] Using TruCOUNT tubes, a known volume of sample is stained directly into the TruCOUNT tube. The lyophilized pellet in the tube dissolves, releasing a known number of fluorescent beads. During analysis, the absolute number of positive cells in the sample (cells / µL) can be determined by comparing cell events to bead events.

[0149] Flow cytometry Cells are vortexed thoroughly (at low speed) to reduce clumping before running on the flow cytometer.

[0150] Data analysis Visually inspect the CD45 vs. SSC dot plot. Lymphocytes appear as bright, compact cell populations with low to moderate SSC. Monocytes (M) and granulocytes (G) appear as distinct populations. The analysis is complete when the monocyte and lymphocyte cell populations show clear separation.

[0151] First, lymphocytes are gated as a CD45-positive, low SSC cell population. CD16 / CD56 versus CD3 is preselected. T cells (T) should appear as compact, bright CD3-positive clusters. NK cells (NK) should appear as compact, bright CD16 / CD56-positive clusters. Gating is now complete, and T cells and NK cells can be counted.

[0152] Bead event counting is performed using a CD16 / CD56 vs. CD3 plot without preselected gates. Beads should appear as PE / FITC double-positive clusters.

[0153] Calculate absolute numbers The absolute number of T cells or NK cells in a sample (cells per µL of blood) is determined by comparing cell events to bead events. Data analysis can be performed using either MultiSET software or manually (using CellQuest or other software). For manual counting, the number of positive cell acquisition events (#) is divided by the number of acquisition bead events (#), then multiplied by (the total number of TruCOUNT beads (lot-dependent) divided by the 50 µL whole blood sample volume). The result is the absolute cell count per microliter. [ka] [ka]

Claims

1. 1. An anti-CD19 antibody for use in treating a patient with a hematological cancer, wherein said patient has a peripheral NK cell count of less than or equal to 100 NK cells / μl at baseline.

2. 2. The anti-CD19 antibody for use in treating patients with hematological cancer according to claim 1, wherein the anti-CD19 antibody comprises an HCDR1 region comprising the sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising the sequence NPYNDG (SEQ ID NO: 2), an HCDR3 region comprising the sequence GTYYYGTRVFDY (SEQ ID NO: 3), an LCDR1 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising the sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6).

3. the anti-CD19 antibody has the sequence The variable heavy chain of EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7) and array 10. An anti-CD19 antibody for use in treating patients with hematological cancer according to any one of the preceding claims, comprising a variable light chain of: DIVMTQSPATHLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK (SEQ ID NO: 8).

4. the anti-CD19 antibody has the sequence EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGT KYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP a heavy chain having CPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11) and array 10. An anti-CD19 antibody for use in treating patients with hematological cancer according to any one of the preceding claims, comprising a light chain having: DIVMTQSPATHLSPGERATTLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12).

5. 10. The anti-CD19 antibody for use in treating a patient with hematological cancer according to any one of the preceding claims, wherein said anti-CD19 antibody is administered in combination with one or more additional agents.

6. 6. The anti-CD19 antibody for use in treating a patient with hematological cancer according to claim 5, wherein the agent is a biological or chemotherapeutic agent, or a pharmaceutically acceptable salt thereof.

7. 7. The anti-CD19 antibody for use in treating a patient with a hematological cancer of claim 6, wherein the one or more agents are a therapeutic antibody or antibody fragment, a nitrogen mustard, a purine analog, a thalidomide analog, a phosphoinositide 3-kinase inhibitor, a BCL-2 inhibitor, a Bruton's tyrosine kinase (BTK) inhibitor, or a pharmaceutically acceptable salt thereof.

8. 8. The anti-CD19 antibody for use in treating a patient with a hematological cancer of claim 7, wherein the one or more agents are rituximab, R-CHOP, cyclophosphamide, chlorambucil, uramustine, ifosfamide, melphalan, bendamustine, mercaptopurine, azathioprine, thioguanine, fludarabine, thalidomide, lenalidomide, pomalidomide, idelalisib, duvelisib, copanlisib, ibrutinib, venetoclax, or a pharmaceutically acceptable salt thereof.

9. 10. The anti-CD19 antibody for use in treating a patient with a hematological cancer according to any one of the preceding claims, wherein said patient has chronic lymphocytic leukemia (CLL), non-Hodgkin's lymphoma (NHL), small lymphocytic lymphoma (SLL) or acute lymphoblastic leukemia (ALL).

10. 10. The anti-CD19 antibody for use in treating a patient with hematological cancer according to claim 9, wherein the patient has non-Hodgkin's lymphoma.

11. 11. The anti-CD19 antibody for use in treating patients with hematological cancer of claim 10, wherein the non-Hodgkin's lymphoma is selected from the group consisting of follicular lymphoma, small lymphocytic lymphoma, mucosa-associated lymphoid tissue, marginal zone lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, and mantle cell lymphoma.

12. 12. The anti-CD19 antibody for use in treating a patient with a hematological cancer according to claim 11, wherein the non-Hodgkin's lymphoma is relapsed or refractory diffuse large B-cell lymphoma (rr DLBCL).

13. The anti-CD19 antibody has the following characteristics: (i) progression-free survival (PFS), (ii) objective response rate (ORR), (iii) duration of response (DoR); (iv) overall survival (OS); (v) increasing one or more of the following: (i) the time to progression (TTP); (ii) the time to progression (TTP);

14. 1. A method for selecting a patient with hematological cancer predicted to benefit from therapeutic administration of an anti-CD19 antibody, said method comprising: a) providing a blood sample obtained from said patient prior to treatment with said anti-CD19 antibody; b) determining the number of peripheral NK cells; c) selecting the patient based on the patient having a peripheral NK cell count of 100 cells / μl or less at baseline.

15. Steps below:

15. The method of claim 14, further comprising: d) treating said selected patient with an anti-CD19 antibody.

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