Sequential Anti-CD19 therapy

Abstract

To provide compositions and methods for sequential treatment of cancer in human patients.SOLUTION: Provided is a therapeutic agent directed against CD19 for use in the treatment of cancer in a patient, the patient being refractory or relapsed from previous treatment, the previous treatment comprising an anti-CD19 antibody comprising an HCDR1 region comprising sequence SYVMH (SEQ ID NO: 1), an HCDR2 region comprising sequence NPYNDG (SEQ ID NO: 2), an HCDR3 region comprising sequence GTYYYGTRVFDY (SEQ ID NO: 3), an LCDR1 region comprising sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), an LCDR2 region comprising sequence RMSNLNS (SEQ ID NO: 5), and an LCDR3 region comprising sequence MQHLEYPIT (SEQ ID NO: 6).SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to immunotherapeutic compositions and methods for the sequential treatment of cancer in human patients using therapeutic agents that bind to human CD19. In particular, immunotherapeutic regimens, including CAR-T cells, are described that are useful for the treatment of B-cell malignancies, such as non-Hodgkin's lymphoma (NHL), including diffuse large B-cell lymphoma (DLBCL), and chronic lymphocytic leukemia (CLL), in patients after prior treatment with the anti-CD19 antibody tafasitamab. [Background technology]

[0002] CD19, a coreceptor for the B cell receptor, is a marker of the B cell lineage because it is expressed throughout B cell development, from differentiation to the final stage of plasma cells (Wang, Exp Hematol Oncol. 2012). CD19 is present on the surface of both healthy and malignant B cells. Most B cell tumors express CD19, including acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), and B cell lymphoma. Approximately 90% of diffuse large B cell lymphomas (DLBCL) express the CD19 antigen (Kimura et al. International Journal of Hematology 2007), compared with approximately 80% of ALL cases. The fact that some tumors do not express CD19 indicates that CD19 is not critical for B cell survival. This is supported by data from CD19 knockout mice, which revealed that CD19 deficiency does not affect the number of early precursor B cells in the bone marrow, nor the size and morphology of B cells. CD19- / - mice instead exhibit a decrease in the total number and frequency of peripheral B cells. Thus, CD19 contributes to the balance between humoral, antigen-induced responses and tolerance induction, and therefore plays a crucial role in initiating optimal immune responses (Wang, Exp Hematol Oncol. 2012).

[0003] Due to its ubiquitous presence on malignant B cells, CD19 is a suitable target for immunotherapy. CD19 expression is restricted to cells of the B lineage and is not found on pluripotent blood stem cells or most other normal tissues (Scheuermann, Leuk Lymphoma. 1995). The major drug classes contributing to the CD19-targeting therapeutic market include monoclonal antibodies (e.g., MOR208, tafasitamab; MEDI-551, inebilizumab), antibody-drug conjugates (e.g., SAR3419, coltuximab ravtansine), bispecific antibodies (e.g., blinatumomab, BLINCYTO®), or chimeric antigen receptor T cell (CAR-T) approaches (e.g., axicabtagene-ciloleucel, YESCARTA®; tisagenlecleucel, KYMRIAH®).

[0004] Tafasitamab is an Fc-enhanced, humanized, monoclonal antibody that targets CD19 and has been shown to exert its antitumor effects through antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and direct cytotoxicity (Awan FT et al. Blood. 2010 Feb 11;115(6):1204-13) (WO2008022152). Tafasitamab recently received FDA Breakthrough Therapy Designation. Currently, tafasitamab is being tested in phase II and III clinical trials in combination with the immunomodulatory agent lenalidomide (L-MIND) and the chemotherapy agent bendamustine (B-MIND) for diffuse large B-cell lymphoma (DLBCL). L-MIND (NCT02399085) is an open-label, single-arm, phase II study of tafasitamab (TAFA) plus lenalidomide (LEN) in patients with relapsed / refractory (R / R) DLBCL who are ineligible for autologous stem cell transplantation (ASCT). B-MIND (NCT02763319) is an open-label, two-arm, phase II / III efficacy and safety study of tafasitamab in combination with bendamustine (BEN) versus rituximab in patients with relapsed / refractory (R / R) DLBCL who are ineligible for high-dose chemotherapy (HDC) and ASCT.

[0005] Adoptive cell transfer (AT) using autologous T cells genetically engineered ex vivo to target tumor antigens is a promising therapeutic approach for the treatment of CD19-positive hematologic malignancies. CD19-directed chimeric antigen receptor T cell (CAR-T) therapy has shown remarkable activity in B-cell lymphoma and acute lymphoblastic leukemia, and two anti-CD19 CAR-T therapies were approved by the FDA in 2017. These anti-CD19 CAR-T cells demonstrated remarkable efficacy in treating patients with relapsed or refractory B-cell lymphomas (Maude S et al., N Engl J Med. 2018, 378:439-48), such as patients with relapsed / relapsed DLBCL after failure of two or more prior systemic therapies (Neelapu S et al., N Engl J Med. 2017, 377:2531-44). Although these trials demonstrated unprecedented efficacy, they also revealed that not all patients respond to anti-CD19 CAR-T cells and that even those who initially respond have a limited duration of response.A patient's ability to respond to CAR-T cell therapy can be influenced, for example, by the number or type of prior treatment regimen, increased expression of inhibitory signals around the CAR-T cells (e.g., PD-L1, which causes suppression), changes in target antigen expression, or epitope loss.

[0006] While other CD19-directed modalities for the treatment of B-cell malignancies have emerged, such as monoclonal antibodies and bispecific T-cell engagers (BiTes), the question of appropriate therapeutic sequencing remains unresolved, and it is unclear whether targeting CD19 with a monoclonal antibody such as tafasitamab will interfere with the ability of CD19-targeted CAR-T to exert antitumor effects in subsequent treatments.

[0007] The high potential of immunotherapy generally comes with the caveat of tumor escape, in which clonal tumor cells develop mechanisms that allow them to resist a particular therapy. One of the earliest resistance mechanisms identified was downregulation of CD19 on the surface of tumor cells (Grupp SA et al, N Engl J Med. (2013) 368:1509-18, Ruella M Comput Struct Biotechnol J. (2016) 14:357-62). Therapeutic compounds exert selective pressure on tumors and malignant clones, but clones that develop resistance to the therapeutic compounds can proliferate. For example, approximately 10% to 20% of acute lymphoblastic leukemia (ALL) patients treated with the CD19 / CD3 bispecific T-cell engager (blinatumomab) experience CD19-negative (CD19-) relapse. Such immune escape can be facilitated by multiple mechanisms, including lineage switching, immunoediting, epitope loss, splice or exon variants, and point mutations, including the acquisition of secondary CD19 mutations that render CD19 nonfunctional. Immune escape is a major form of treatment resistance in ALL patients (Braig F et al., 2017 Blood. 129(1):100-104; Grupp SA et al. N Engl J Med. 2013 Apr 18;368(16):1509-1518). Targeting specific antigens on the tumor cell surface with monoclonal antibodies allows for the selection of clones that are not recognized by or are not affected by antibody binding.

[0008] In such cases, a key question to be answered in the era of targeted immunotherapy is: Is it possible to target the same tumor antigen with a different cancer immunotherapy modality after disease progression following prior therapy against the same antigen? In such clinical scenarios, concerns exist regarding the persistence of antigen blockade from the prior treatment regimen. There are also concerns regarding the selective pressure of the prior treatment targeting a specific antigen allowing the progression of clones that do not express that antigen, rendering subsequent treatment directed against the same target ineffective (antigen escape). Therefore, the development of novel approaches that attenuate, block, or eliminate antigen-loss escape or that do not have any interference with prior treatment regimens directed against the same antigen would be an advancement in this field.

[0009] It is known from the prior art that CD19 can be readily internalized upon Ab binding (Pulczynski S Blood. 1993, 81(6):1549-57), and that loss of CD19 expression on tumor cells is a frequent escape mechanism in patients treated with CD19-targeted T cells (Grupp SA, N Engl J Med. 2013 Apr 18;368(16):1509-1518). Dual targeting with CD19 and CD123 has been described to prevent antigen loss recurrence after CD19-directed immunotherapy (Ruella et al. J Clin Invest. 2016 Oct 3;126(10):3814-3826), but this concept uses a combination of multiple targets.

[0010] The present invention is based on the surprising finding that CD19 can remain a target after treatment with the monoclonal anti-CD19 antibody tafasitamab (MOR208). Therefore, another, optionally different, anti-CD19 moiety can be administered after tafasitamab treatment. Specifically, the inventors of the present disclosure surprisingly found that there is no functional interference between tafasitamab and CD19-directed CAR-T cells (CART19). This finding is supported by an unpublished case study demonstrating sustained remission in a patient with relapsed and refractory DLBCL achieved with anti-CD19 CAR-T cell therapy after prior treatment with tafasitamab as part of the L-MIND clinical trial. Summary of the Invention

[0011] Disclosed are methods and compositions for treating hematological cancers, comprising administering to a patient a composition comprising autologous T cells, the autologous T cells expressing a recombinant receptor that specifically binds to an antigen that is specific for the hematological cancer, and the patient having previously been treated with a composition comprising an anti-CD19 antibody, the antibody comprising: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0012] More particularly, one embodiment relates to methods and compounds for treating CD19+ hematological cancers. Specifically, the antigen specific to this hematological cancer is human CD19. Preferably, the CD19+ hematological cancer is DLBCL. In another preferred embodiment, the DLBCL is relapsed or refractory (R / R) DLBCL.

[0013] In one embodiment, the autologous T cells expressing a recombinant receptor are CAR-T cells. In one embodiment, the CAR-T cells are directed against CD20 or CD19. In a preferred embodiment, the CAR-T cells are directed against CD19. More particularly, the CAR-T cells are directed against CD19 and are selected from the group consisting of axicabtagene ciloleucel (YESCARTA®), tisagenlecleucel (KYMRIAH®), lysocabtagene maraleucel (JCAR017), and UCART19.

[0014] In particular, CD19-directed CAR-T cells are disclosed for use in treating cancer in patients who have been previously treated with an anti-CD19 antibody.

[0015] More specifically, one embodiment relates to CD19-directed CAR-T cells for use in treating cancer in a patient, where the patient has been previously treated with a composition comprising tafasitamab. In a specific embodiment, tisagenlecleucel CAR-T cells are disclosed for use in treating cancer in a patient, where the patient has been previously treated with a composition comprising tafasitamab. A specific embodiment relates to CD19-directed CAR-T cells for use in treating R / R DLBCL in a patient, where the patient has been previously treated with a composition comprising tafasitamab. Another aspect relates to CD19-directed CAR-T cells for use in treating R / R DLBCL in a patient, where the patient has been previously treated with a combination therapy comprising tafasitamab and lenalidomide. In a specific embodiment, tisagenlecleucel CAR-T cells are disclosed for use in treating R / R DLBCL in a patient, where the patient has been previously treated with a composition comprising tafasitamab. In another aspect, disclosed are tisagenlecleucel CAR-T cells for use in treating R / R DLBCL in a patient, the patient having been previously treated with a combination therapy comprising tafasitamab and lenalidomide. [Brief explanation of the drawings]

[0016] [Figure 1] Cytotoxicity of tafasitamab against the Jeko-1 cell line (A) with and without natural killer (NK) cells. CD19+, luciferase+ Jeko cells were incubated with various concentrations of tafasitamab in the presence or absence of NK cells, and cytotoxicity was assessed 24 hours later. In the presence of NK cells, tafasitamab demonstrates significantly enhanced antitumor cytotoxicity. Activity was determined by bioluminescence imaging after 24 hours. (n=3 independent experiments; data were consistent with those targeting Nalm-6 and Ly7 cell lines (B and C). [Figure 2]Clone FMC63 (CD19-binding domain of CART19) and tafasitamab compete for CD19 binding. Jeko cells were treated with 2 or 0.5 mg / mL tafasitamab for 4 hours, followed by flow cytometry using the FMC63 anti-CD19 antibody. A significantly reduced CD19 signal was observed by flow cytometry, indicating that tafasitamab and FMC63 directly compete for binding. [Figure 3] CART19 exhibits potent antigen-specific cytotoxicity in the presence of the monoclonal anti-CD19 antibody tafasitamab. Luciferase-positive, CD19+ Jeko cells (A) were incubated with various concentrations of tafasitamab, and then 3 hours later, CART19 cells were added at various E:T ratios as indicated. Activity was determined by bioluminescence imaging 24 hours later (n=3 independent experiments). Consistent results were obtained using Ly7 or Nalm-6 cell lines (B and C). [Figure 4] CART19 cells continue to exhibit significant antigen-specific degranulation and cytokine production in the presence of the anti-CD19 antibody tafasitamab. When CART19 cells or untransduced T cells (UTD) were incubated with Jeko at a 1:5 ratio (with or without pretreatment with tafasitamab) and in the presence of CD28, CD49d, and monensin (according to standard degranulation methods), CART19 cells continued to exhibit similar levels of degranulation and cytokine production despite CD19 binding to the monoclonal antibody (n = 2 independent experiments). GM-CSF, INFγ, IL-2, and MIP1b cytokine production levels did not differ between the presence and absence of tafasitamab (data not shown). [Figure 5] CART19 cells exhibit significant antigen-specific proliferation that is not impaired by the presence of tafasitamab. When CART19 or UTD were cocultured with CD19+ Jeko cells at a 1:1 ratio in the presence or absence of tafasitamab, CART19, but not UTD, exhibited significant antigen-specific proliferation that was not different from when CD19 was blocked with tafasitamab. [Figure 6]DLBCL case study: Anti-CD19 CAR-T cell therapy is feasible after tafasitamab plus lenalidomide [Figure 7] On day -14, NGS mice were injected with JeKo-1 cells to induce tumors. On day -8, mice were randomly divided into a tafasitamab group (10 mice) and a PBS group (4 mice, Δ). The tafasitamab group received tafasitamab at 10 mg / kg three times weekly via intravenous injection. On day -1, mice receiving tafasitamab were randomly divided into tafasitamab continuation (○) or tafasitamab discontinuation (□) groups. On day 0, all mice in the three groups were injected with 2.5 x 10 CART19 cells (IV). The tafasitamab continuation group showed reduced survival compared with the tafasitamab discontinuation group or the PBS control (**p=0.005, log-rank test, continuation vs. discontinuation, or continuation vs. PBS; no significant difference between the PBS group and the tafasitamab discontinuation group). [Figure 8] On day -14, NGS mice were injected with JeKo-1 cells to induce tumors. On day -8, mice were randomly divided into a tafasitamab group (10 mice) and a PBS group (4 mice, Δ). The tafasitamab group received tafasitamab at 10 mg / kg ip injection three times weekly. On day -1, mice receiving tafasitamab were randomly divided into tafasitamab continuation (○) or tafasitamab discontinuation (□) groups. On day 0, all mice in the three groups were injected with 2.5 x 10 CART19 cells (IV). (A) The tafasitamab continuation group showed higher tumor burden than the tafasitamab discontinuation group or the PBS control (****p<0.0001, two-way ANOVA, continuation vs. discontinuation, or continuation vs. PBS). (B) No difference was observed between the tafasitamab discontinuation group and the PBS group (ns not significant, two-way ANOVA, discontinuation vs. PBS). DETAILED DESCRIPTION OF THE INVENTION

[0017] definition The term "antibody" refers to monoclonal antibodies of any isotype, e.g., IgG, IgM, IgA, IgD, and IgE. IgG antibodies are composed of two identical heavy chains and two identical light chains, which are held together by disulfide bonds. Each heavy and light chain contains a constant region and a variable region. Each variable region contains three segments called "complementarity-determining regions" ("CDRs") or "hypervariable regions," which are primarily responsible for binding to an epitope on an antigen. These segments are numbered sequentially from the N-terminus and designated CDR1, CDR2, and CDR3. The more highly conserved portions of the variable region outside the CDRs are called "framework regions." An "antibody fragment" refers to an Fv, scFv, dsFv, Fab, Fab', F(ab')2 fragment, or other fragment, which contains at least one variable heavy or light chain, each chain containing a CDR and framework region.

[0018] "VH" refers to the variable region of the immunoglobulin heavy chain of an antibody or antibody fragment. "VL" refers to the variable region of the immunoglobulin light chain of an antibody or antibody fragment.

[0019] The term "CD19" refers to the protein known as CD19, which 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. Human CD19 (UniProt-P15391) has the following amino acid sequence: (SEQ ID NO: 13). Variants of human CD19 (e.g., splice variants, genetic polymorphisms, and SNPs) are also encompassed by the present application.

[0020] "MOR00208," "MOR208," "XmAb5574," or "tafasitamab" is an anti-CD19 antibody. The amino acid sequences of the VH and VL domains are set forth in SEQ ID NO: 7 and SEQ ID NO: 8, respectively. The amino acid sequence of the MOR208 heavy chain Fc region is as follows: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 9). The amino acid sequence of the MOR208 light chain Fc region is as follows: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 10). The MOR208 antibody is described in U.S. Patent Application Serial No. 12 / 377,251, which is incorporated by reference in its entirety, as an antibody named 4G7 H1.52 Hybrid S239D / I332E / 4G7 L1.155 (subsequently named MOR00208), and has the following sequence: >4G7 H1.52 Hybrid S239D / I332E EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKALPAPEEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 11) >4G7 L1.155 DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 12)

[0021] "Administered" or "administration" includes, but is not limited to, drug delivery in an injectable form, e.g., by intravenous, intramuscular, intradermal, or subcutaneous routes, or via mucosal routes, e.g., as a nasal spray or aerosol for inhalation, or as an orally ingestible liquid, capsule, or tablet. Preferably, administration is in an injectable form.

[0022] The term "chimeric antigen receptor (CAR)" as used herein can refer to, for example, an engineered T cell receptor, a chimeric T cell receptor, or a chimeric immune receptor, and can encompass engineered receptors with defined specificity grafted onto specific immune effector cells. CARs can be used to confer the specificity of a monoclonal antibody to T cells, thereby enabling the generation of large numbers of specific T cells, for example, for use in adoptive cell therapy. In specific embodiments, CARs direct the specificity of cells to, for example, a tumor-associated antigen. In some embodiments, CARs comprise an intracellular activation domain, a transmembrane domain, and an extracellular domain comprising a tumor-associated antigen-binding region. In some aspects, CARs comprise a fusion of a monoclonal antibody-derived FMC63 single-chain variable fragment (scFv) fused to a hinge linker CD8h, a transmembrane domain CD8TM, and a signaling endodomain 41BBζ. The specificity of the CAR design can be derived from an antibody fragment (e.g., scFv, Fab, VHH, scFab), a receptor ligand (e.g., a peptide), or a dectin. In a specific embodiment, malignant B cells can be targeted by redirecting T cell specificity using a CAR specific for the B cell lineage molecule, CD19. In certain cases, the CAR contains additional domains for costimulatory signaling, such as, but not limited to, CD3ζ, FcR, CD27, CD28, CD137, DAP10, 41BBζ, and / or OX40. In some cases, molecules can be co-expressed with the CAR, including costimulatory molecules, reporter genes for imaging (e.g., for positron emission tomography), gene products that conditionally ablate T cells upon addition of a prodrug, homing receptors, cytokines, and cytokine receptors.

[0023] A "therapeutically effective amount" of a compound or combination refers to an amount sufficient to cure, alleviate, or partially arrest the clinical symptoms of a given disease or disorder and its complications. The amount that is 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. Of course, determining appropriate dosages can be accomplished using routine experimentation, by constructing a matrix of values ​​and examining the matrix at different time points, all of which is within the ordinary skill of a skilled physician or clinical scientist.

[0024] 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.

[0025] Non-Hodgkin's lymphoma (NHL) is a heterogeneous malignant tumor of lymphocyte origin. In the United States, its incidence is estimated at 65,000 cases per year, with approximately 20,000 deaths (American Cancer Society, 2006; and SEER Cancer Statistics Review). The disease can occur at any age, but onset typically begins in adults over the age of 40, and incidence increases 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 physicians is the World Health Organization (WHO) Classification of Tumors, which organizes NHL into precursor and mature B-cell or T-cell neoplasms. PDQ currently categorizes NHL as indolent or aggressive for clinical trial participation. The indolent NHL group is primarily composed of follicular subtypes, small lymphocytic lymphoma, mucosa-associated lymphoid tissue (MALT), and marginal zone lymphoma; indolent NHL accounts for approximately 50% of newly diagnosed B-cell NHL patients. Aggressive NHL includes patients with the histological diagnosis of primary diffuse large B-cell lymphoma (DLBL, "DLBCL," or DLCL) (40% of all newly diagnosed patients have diffuse large B-cell lymphoma), Burkitt's lymphoma, and mantle cell lymphoma ("MCL"). The most commonly used agents in combination chemotherapy include cyclophosphamide, vincristine, and prednisone (CVP); or cyclophosphamide, adriamycin, vincristine, and prednisone (CHOP). Approximately 70% to 80% of patients respond to their initial chemotherapy, with remission periods lasting approximately 2 to 3 years. Ultimately, the majority of patients relapse. The discovery and clinical use of the anti-CD20 antibody rituximab has led to significant improvements in response and survival. 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 multiple types of NHL and is currently approved as first-line treatment for both indolent (follicular lymphoma) and aggressive NHL (diffuse large B-cell lymphoma). However, anti-CD20 monoclonal antibodies (mAbs) have significant limitations, including primary resistance (50% response in relapsed indolent patients), acquired resistance (50% response rate upon retreatment), rare complete responses (2% complete response rate in the relapsed population), and sequential patterns of relapse. Finally, many B cells do not express CD20, and therefore, many B-cell disorders cannot be treated with anti-CD20 antibody therapy.

[0026] Chronic lymphocytic leukemia (also known as "lymphoid leukemia" or "CLL") is a type of adult leukemia caused by the abnormal accumulation of B lymphocytes. In CLL, the malignant lymphocytes may appear normal and mature, but they are unable to effectively fight 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 a mortality rate of approximately 5,000 cases per year (American Cancer Society, 2006; and SEER Cancer Statistics Review). CLL is incurable, but its progression is indolent in most cases. Many people with CLL lead normal, active lives for many years. Due to its slow onset, early-stage CLL is generally not treated, as early CLL intervention is not believed to improve survival or quality of life. Instead, the condition is monitored over time. Initial CLL treatment varies depending on the exact diagnosis and progression of the disease. There are many agents used to treat CLL. Combination chemotherapy regimens, such as FCR (fludarabine, cyclophosphamide, and rituximab) and BR (ibrutinib and rituximab), are effective for both newly diagnosed and relapsed CLL. Allogeneic bone marrow (stem cell) transplantation is rarely used as a first-line treatment due to its risks.

[0027] 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 common variant of CLL with common pathological and immunophenotypic features (Campo et al., 2011). The definition of SLL requires the presence of lymphadenopathy and / or splenomegaly. Furthermore, the peripheral blood B lymphocyte count must not exceed 5E+09 / L. In SLL, the diagnosis should be confirmed by histopathological evaluation of lymph node biopsy whenever possible (Hallek et al., 2008). The incidence of SLL is approximately 25% of CLL in the United States (Dores et al., 2007).

[0028] 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 and immature white blood cells (also known as lymphoblasts) in the bone marrow. The "acute" refers to the undifferentiated, immature state of the circulating lymphocytes ("blasts") and the rapid progression of the disease, with a life expectancy of a few weeks to a few months if left untreated.

[0029] "Subject" or "patient," as used in this context, refers to any mammal, including rodents, such as mice or rats, and primates, such as cynomolgus monkeys (Macaca fascicularis), rhesus monkeys (Macaca mulatta), or humans (Homo sapiens). Preferably, the subject or patient is a primate, particularly preferably a human.

[0030] The terms "combination" or "pharmaceutical combination" refer to the administration of one treatment in addition to another treatment. Thus, "in combination with" includes simultaneous (e.g., concurrent) and consecutive administration in any order. By way of example and not limitation, a first treatment (e.g., an agent such as an anti-CD19 antibody) can be administered (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, 12 weeks, 14 weeks, 16 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 hours, 74 hours, 75 hours, 7 The compound can be administered to the patient at or after (e.g., 1 week, 11 weeks, or 12 weeks before), concurrently with, 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, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks or more after).

[0031] The terms "sequential combination" or "sequential treatment combination" refer to the administration of a different treatment after a prior treatment has been completed. An additional treatment can be included between the two treatments. By way of example, and in embodiments as disclosed herein, sequential combination refers to treating a patient with chimeric antigen receptor (CAR) T cells directed against CD19 after the patient has relapsed from or is refractory to a prior treatment comprising an anti-CD19 antibody as disclosed herein. In certain embodiments, the prior or first treatment of the sequential combination is completed 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks or more before the second, different treatment is administered.

[0032] "Thalidomide analogs" include, but are not limited to, thalidomide itself, lenalidomide (CC-5013, Revlimid™), pomalidomide (CC4047, Actimid™), and compounds disclosed in WO2002068414 and WO2005016326, both of which are incorporated by reference in their entirety. This term refers to chemically synthesized compounds that use the thalidomide structure as a backbone (e.g., to which side chains 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, one or more functional groups, or charge upon ionization. The term "thalidomide analog" also includes metabolites of thalidomide. Thalidomide analogs include racemic mixtures of the S and R enantiomers of each compound, as well as the individual S or R enantiomers. Racemic mixtures are preferred. Thalidomide analogs include compounds such as lenalidomide, which has the following structure: [ka]

[0033] The term "recurrence," as used herein, refers to the reappearance of a disease (e.g., cancer) after an initial period of response (e.g., complete or partial response), e.g., after prior treatment with a therapy, e.g., cancer treatment. More generally, in certain embodiments, a response (e.g., complete or partial response) may involve the absence of detectable MRD (minimal residual disease). In certain embodiments, the initial period of response lasts for at least 1, 2, 3, 4, 5, or 6 days; at least 1, 2, 3, or 4 weeks; at least 1, 2, 3, 4, 6, 8, 10, or 12 months; or at least 1, 2, 3, 4, or 5 years.

[0034] "Refractory," as used herein, refers to a disease, e.g., cancer, that does not respond to treatment. In embodiments, a refractory cancer may be resistant to treatment before or at the start of treatment. In other embodiments, a refractory cancer may become resistant during treatment. A refractory cancer is also referred to as a resistant cancer. [Table 1-1] [Table 1-2] [Table 1-3]

[0035] In various embodiments, the invention pertains to nucleic acid sequences encoding any of the disclosed antibodies or CARs for use in treating cancer in a patient as described herein. [Example]

[0036] The CART19 cells used in these experiments were generated by lentiviral transduction of healthy donor T cells with a second-generation CD19CAR construct (clone FMC63, FMC63-CD8h-CD8TM-41BBζ, similar to the FDA-approved construct used in tisagenlecleucel, containing the CD19-binding domain). T cells isolated from healthy donors were stimulated with Cell Therapy Systems Dynabeads CD3 / CD28 (Life Technologies, Oslo, Norway) at a 1:3 ratio (cells:beads) and then transduced with lentiviral particles at an MOI of 3.0 24 hours after stimulation. On day 6, magnetic beads were removed and T cells were assessed for CAR19 expression by flow cytometry. On day 8, CART cells were collected and cryopreserved for future experiments. CART cells were thawed and rested in T cell medium 6–12 hours before their use in experiments, as specified in each experiment.

[0037] Example 1 The functional activity of tafasitamab or CART19 cells was tested in the CD19-positive target cell lines Jeko (mantle cell lymphoma), Ly7 (DLBCL), and Nalm-6 (ALL). Cell lines were originally obtained from ATCC or DSM-Z. Cell lines were transduced with luciferase (firefly / EGFP, CBG / EGFP, or CBR / EGFP) and then sorted to obtain >99% positive populations. Clear activity was observed for both treatments in 24-hour ADCC assays (tafasitamab titration + natural killer (NK) cells; Figure 1) and T cell cytotoxicity assays (CART19, E:T titration; data not shown) in all cell lines tested. Killing assays were performed according to standard protocols (CaoL-F et al. Cytometry A, 2010, vol. 77 6 (pg.534-545)). Briefly, cytotoxicity was assessed after 24 hours of incubation of CD19+ luciferase+ target cells with various concentrations of tafasitamab in the presence or absence of NK cells.

[0038] Example 2 We investigated whether the observed CART19 activity might be affected by tafasitamab if there is direct CD19 binding competition between tafasitamab and CAR. To first test for such binding competition, we incubated the CD19+ cell lines Nalm-6 or Jeko with 2 or 0.5 mg / mL tafasitamab to saturate the receptor. Subsequent flow cytometry analysis using the FMC63 antibody (which possesses the same CD19-binding domain as CART19) failed to detect CD19 expression, indicating direct binding competition between FMC63 and tafasitamab (Figure 2). To examine the potential impact of such binding competition on CART19 cell effector function, increasing concentrations of tafasitamab were incubated with target cells (CD19+ cell lines, Jeko, Ly7, or Nalm-6), and then CART19 cells were added at different effector:target ratios (0.1:1 to 10:1) without other effector cells in the cell culture. The presence of tafasitamab, i.e., binding to the CD19 antigen, did not affect important CART cell effector functions, such as antigen-specific killing (Figure 3), degranulation (Figure 4), cytokine production, or antigen-specific proliferation of CART19 cells (Figure 5).

[0039] In summary, the data show that despite the presence of tafasitamab and its competition for CD19 binding, CART19 cells continue to exhibit potent antigen-specific effector function: targeting CD19 with tafasitamab does not impair CD19-directed chimeric antigen receptor (CAR) T-cell activity in vitro.

[0040] Example 3 In a case study of a 58-year-old female patient who initially presented with an 8-cm mesenteric mass, workup revealed stage III germinal center B-cell-like (GCB) DLBCL arising from follicular lymphoma, a Ki-67 proliferation index of 80%, and an IGH / BCL2 fusion. The patient received six cycles of dose-adjusted R-EPOCH. Although a complete remission (CR) was achieved with this state-of-the-art treatment, the patient experienced a relapse within two years. Subsequently, the patient received rituximab plus ifosfamide, carboplatin, and etoposide (RICE) chemotherapy, which achieved a second CR. The patient refused ASCT. A second relapse occurred approximately two years later. After meeting the eligibility criteria, the patient was enrolled in the L-MIND clinical trial and received six cycles of TAFA plus LEN (1 cycle = 28 days; TAFA at 12 mg / kg intravenously once weekly for 3 cycles, followed by LEN at 25 mg daily on days 1–21 of each cycle). TAFA / LEN was well tolerated. Stable disease was achieved with this investigational regimen, but progression occurred after 6 months. The fourth treatment consisted of four cycles of rituximab, gemcitabine, and oxaliplatin (R-GEM-OX), to which the patient demonstrated a partial response. Shortly thereafter, the patient received CD19-directed CART therapy (axicabtagene-ciloleucel [YESCARTA®]) (Figure 6). The course of treatment was complicated by grade 2 cytokine release syndrome. A complete response was achieved one month after treatment and has remained so ever since. As of the current data, patients remain free of clinical evidence of relapse.Sustainable remissions have been achieved with CD19-directed CAR-T cell therapy in patients with relapsed and refractory diffuse large B-cell lymphoma (R / R-DLBCL) despite prior treatment with tafasitamab.

[0041] The half-life of tafasitamab is approximately 16 days, suggesting that tafasitamab was eliminated before the infusion of CAR-T cells 5 months later. Although a biopsy was not performed at the time of progression after TAFA, CD19 antigen escape is unlikely to have been the primary cause of relapse, as the patient achieved a durable remission with subsequent CD19-directed CAR-T therapy. Therefore, disease progression after treatment with the anti-CD19 monoclonal antibody tafasitamab does not preclude a patient from CD19-directed CAR-T therapy, even if the same antigen was previously targeted.

[0042] Example 4 On day −14, NSG (NOD scid gamma mice, immunodeficient experimental mice) mice were treated with 1 × 10 6 On day -8, tumor burden was assessed by bioluminescence imaging (BLI), and mice were randomly assigned to a tafasitamab group (10 mice) or a PBS group (4 mice). The tafasitamab group received 10 mg / kg ip injection of tafasitamab three times a week. On day -1, tumor burden was assessed by BLI, and mice were randomly assigned to either a tafasitamab continuation or tafasitamab discontinuation group. On day 0, all mice in the three groups received 2.5 x 10 6 Mice were administered CART19 cells (IV). Mice in the tafasitamab continuation group continued tafasitamab treatment in parallel with CART19 cells. All mice were monitored weekly by BLI to monitor tumor burden and survival. Mice were then sacrificed.

[0043] CD19 competition was observed in the tafasitamab-continued group, demonstrating shorter survival (p=0.005, log-rank test, continued vs. discontinued or continued vs. PBS). However, in the tafasitamab-discontinued group, mice achieved remission and did not experience a decrease in survival. There was no difference in overall survival between the PBS and tafasitamab-discontinued groups (log-rank test) (Figure 7).

[0044] Furthermore, the tafasitamab continuation group showed a higher tumor burden compared to the discontinuation group or the PBS control group ( **** p<0.0001, two-way ANOVA, continuation vs. discontinuation or continuation vs. PBS). No difference was observed between the tafasitamab-discontinued group and the PBS group (ns not significant, two-way ANOVA, discontinuation vs. PBS) (Figure 8).

[0045] Therefore, these in vivo data indicate that CD19-directed CAR-T cell therapy is not impaired by prior tafasitamab treatment. We can conclude that prior tafasitamab treatment does not preclude patients from CD19-directed CAR-T cell therapy.

[0046] However, these data also indicate that combined co-treatment of a CD19 antibody and a CD19-directed CAR-T cell therapy may negatively impact the efficacy of the therapy if the CD19 antibody and CD19 CART compete for the same or overlapping CD19 epitope.

[0047] Embodiment In one embodiment, a CD19-directed therapeutic agent is disclosed for use in treating cancer in a patient, the patient having been previously treated with a composition comprising an anti-CD19 antibody, the antibody having 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).

[0048] In one embodiment, a CD19-directed therapeutic agent is disclosed for use in treating cancer in a patient who is refractory to or has experienced a relapse after a prior treatment, the prior treatment comprising an anti-CD19 antibody having 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).

[0049] In another embodiment, a CD19 directed therapeutic is disclosed for use in treating cancer in a patient, wherein the patient has experienced resistance or relapse to a prior treatment, wherein the prior treatment comprises a compound selected from the group consisting of: EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVTVSS (SEQ ID NO: 7) and / or has a variable heavy chain comprising the sequence DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKLEIK (SEQ ID NO: 8) and an anti-CD19 antibody having a variable light chain comprising:

[0050] In one embodiment, a CD19-directed therapeutic agent is disclosed for use in treating cancer in a patient who is refractory to or has experienced a relapse following a prior treatment, the prior treatment comprising an anti-CD19 antibody having a variable heavy chain and / or variable light chain with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the variable heavy chain of SEQ ID NO:7 and / or the variable light chain of SEQ ID NO:8.

[0051] In a preferred embodiment, a CD19-directed therapeutic agent is disclosed for use in treating cancer in a patient who is refractory to or has experienced a relapse following a prior treatment, the prior treatment comprising an anti-CD19 antibody, the antibody having a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12.

[0052] In another preferred embodiment, a CD19-directed therapeutic is disclosed for use in treating cancer in a patient, wherein the patient is refractory to or has experienced a relapse following a prior treatment, wherein the prior treatment comprises the anti-CD19 antibody tafasitamab.

[0053] In another embodiment, a CD19-directed therapeutic is disclosed for use in treating cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising an anti-CD19 antibody that competes with the CD19-directed therapeutic for binding to human CD19.

[0054] In other embodiments, a CD19-directed therapeutic is disclosed for use in treating cancer in a patient, wherein the patient is refractory to or has experienced a relapse after a prior treatment, wherein the prior treatment comprises an anti-CD19 antibody, and wherein the CD19-directed therapeutic is selected from the group of an antibody, an antibody-drug conjugate, a bispecific antibody, a surrogate scaffold protein, or a chimeric antigen receptor (CAR) T cell.

[0055] In a preferred embodiment, the CD19-directed therapeutic agent for use in treating cancer in a patient who is refractory to or has relapsed from prior treatment with an anti-CD19 antibody is a chimeric antigen receptor (CAR) T cell. In certain embodiments, the CAR T cell is preferably one of axicabtagene-ciloleucel (KTE-C19, Axi-cel, YESCARTA®) and / or tisagenlecleucel (CTL019, KYMRIAH®), lisocabtagene-malaleucel (JCAR017, clinical trials: e.g., NCT02631044, NCT03310619), or UCART19 (S68587, NCT02808442), or a combination of two or more thereof. In another embodiment, the CAR-T cells are one of the CAR-T cell constructs known as KITE037, welgenaleucel, ICTCAR-003, IM-19, CTX-110, SSCAR-010, ICTCAR-011, or a combination of two or more thereof.

[0056] In other embodiments, a therapeutic agent for use in treating cancer in a patient is directed against a B-cell lineage marker, and the patient has become refractory or relapsed to a prior treatment, wherein the prior treatment comprised an anti-CD 19 antibody. Preferably, a therapeutic agent for use in treating cancer in a patient is directed against CD19 and / or CD20, and the patient has become refractory or relapsed to a prior treatment, wherein the prior treatment comprised the anti-CD 19 antibody tafasitamab.

[0057] In another embodiment, the CAR-T cells for use in treating cancer in a patient are directed against CD19 and / or CD20, and the patient is refractory to or has relapsed from prior treatment, wherein the prior treatment comprises the anti-CD19 antibody tafasitamab.

[0058] In a preferred embodiment, the therapeutic agent is for use in a pharmaceutical composition. In another embodiment, the therapeutic agent is included in a pharmaceutical composition.

[0059] In one aspect, a CD19-directed therapeutic agent is disclosed for use in treating cancer in a patient, the patient being refractory or experiencing relapse to a prior treatment, the prior treatment including an anti-CD19 antibody, and the cancer being a hematological cancer. In certain embodiments, the hematological cancer is selected from non-Hodgkin's B-cell lymphomas, such as follicular lymphoma (FL), small lymphocytic lymphoma (SLL), Burkitt's lymphoma, and diffuse large B-cell lymphoma (DLBCL), Waldenstrom's hypergammaglobulinemia, leukemia, such as chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), or acute myeloid leukemia (AML). In a preferred embodiment, the hematological cancer is DLBCL. In other embodiments, the cancer is a relapsed or refractory (R / R) cancer, preferably a relapsed or refractory hematological cancer. The R / R hematological cancer is selected from R / R non-Hodgkin's B-cell lymphomas, such as R / R follicular lymphoma (FL), R / R small lymphocytic lymphoma (SLL), R / R Burkitt's lymphoma, and R / R diffuse large B-cell lymphoma (DLBCL), R / R Waldenstrom's hypergammaglobulinemia, R / R leukemias, such as R / R chronic lymphocytic leukemia (CLL), R / R acute lymphoblastic leukemia (ALL), or R / R acute myeloid leukemia (AML). In a preferred embodiment, the R / R hematological cancer is relapsed or refractory (R / R) diffuse large B-cell lymphoma (DLBCL).

[0060] In certain embodiments, CD19-directed CAR-T cells are disclosed for use in treating R / R DLBCL in a patient who is refractory or has experienced a relapse to a prior treatment, wherein the prior treatment comprises an anti-CD19 antibody, preferably an antibody having i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0061] In certain embodiments, CD19-directed CAR-T cells are disclosed for use in treating R / R DLBCL in a patient who is refractory or has relapsed to a prior treatment, wherein the prior treatment comprises a combination of tafasitamab and lenalidomide.

[0062] In another embodiment, CD19-directed CAR-T cells are disclosed for use in treating R / R DLBCL in a patient who is refractory or has relapsed to a prior treatment, wherein the prior treatment comprises a combination of tafasitamab and bendamustine.

[0063] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is treated with CAR-T cells directed against CD19 after the anti-CD19 antibody treatment, and the anti-CD19 antibody has 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).

[0064] In one embodiment, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is treated with CAR-T cells directed against CD19 after the anti-CD19 antibody treatment, and the anti-CD19 antibody has a variable heavy chain of SEQ ID NO:7 and a variable light chain of SEQ ID NO:8.

[0065] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is treated with CAR-T cells directed against CD19 after the anti-CD19 antibody treatment, and the anti-CD19 antibody has a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12.

[0066] In certain embodiments, disclosed herein are anti-CD19 antibodies for use in treating cancer in a patient, wherein the patient is treated with CAR-T cells directed against CD19 after the anti-CD19 antibody treatment, and the anti-CD19 antibody comprises tafasitamab.

[0067] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is refractory or has relapsed after the anti-CD19 antibody treatment and is treated with CAR-T cells directed against CD19, and wherein the anti-CD19 antibody has 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).

[0068] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is refractory or has relapsed after the anti-CD19 antibody treatment and is treated with CAR-T cells directed against CD19, and wherein the anti-CD19 antibody has a variable heavy chain of SEQ ID NO:7 and a variable light chain of SEQ ID NO:8.

[0069] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is refractory or has experienced a relapse after treatment with the anti-CD19 antibody and is treated with CAR-T cells directed against CD19, and wherein the anti-CD19 antibody has a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12.

[0070] In certain aspects, disclosed herein are anti-CD19 antibodies for use in treating cancer in a patient, wherein the patient is refractory to or has experienced a relapse after treatment with the anti-CD19 antibody and is treated with CAR-T cells directed against CD19, and wherein the anti-CD19 antibody comprises tafasitamab.

[0071] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is refractory or has relapsed after treatment with the anti-CD19 antibody in combination with lenalidomide or bendamustine, and is being treated with CAR-T cells directed against CD19, and wherein the anti-CD19 antibody has 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).

[0072] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is refractory or has relapsed following treatment with the anti-CD19 antibody in combination with lenalidomide or bendamustine, and is treated with CAR-T cells directed against CD19, and wherein the anti-CD19 antibody has a variable heavy chain of SEQ ID NO:7 and a variable light chain of SEQ ID NO:8.

[0073] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is refractory or has relapsed after treatment with the anti-CD19 antibody in combination with lenalidomide or bendamustine, and is treated with CAR-T cells directed against CD19, and the anti-CD19 antibody has a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12.

[0074] In certain aspects, disclosed herein are anti-CD19 antibodies for use in treating cancer in a patient, wherein the patient is refractory or has relapsed following treatment with the anti-CD19 antibody in combination with lenalidomide or bendamustine, and is treated with CD19-directed CAR-T cells, and the anti-CD19 antibody comprises tafasitamab.

[0075] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient, wherein the patient is refractory or has experienced a relapse after treatment with the anti-CD19 antibody and has been treated with CAR-T cells directed against CD19, and the anti-CD19 antibody has 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 HCDR4 region comprising the sequence RSSKSLQNVNGNTYLY (SEQ ID NO: 4), and a CAR-T cell directed against CD19. The CD19-directed CAR-T cells have an LCDR1 region comprising the sequence RMSNLNS (SEQ ID NO: 5), an LCDR2 region comprising the sequence MQHLEYPIT (SEQ ID NO: 6), and the CD19-directed CAR-T cells comprise one of axicabtagene-ciloleucel (YESCARTA®) and / or tisagenlecleucel (KYMRIAH®), lysocabtagene-malaleucel (JCAR017), or UCART19 (S68587), or a combination of two or more of these.

[0076] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient who is refractory or has relapsed after anti-CD19 antibody treatment and is treated with CD19-directed CAR-T cells, and the anti-CD19 antibody has a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, and the CD19-directed CAR-T cells comprise one of axicabtagene ciloleucel (YESCARTA®) and / or tisagenlecleucel (KYMRIAH®), lysocabtagene maraleucel (JCAR017), or UCART19 (S68587), or a combination of two or more thereof.

[0077] In one aspect, disclosed herein is an anti-CD19 antibody for use in treating cancer in a patient who is refractory or has relapsed after anti-CD19 antibody treatment and is treated with CD19-directed CAR-T cells, and the anti-CD19 antibody has a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, and the CD19-directed CAR-T cells comprise one or a combination of two or more of axicabtagene ciloleucel (YESCARTA®) and / or tisagenlecleucel (KYMRIAH®), lysocabtagene maraleucel (JCAR017), or UCART19 (S68587).

[0078] In certain aspects, disclosed herein are anti-CD19 antibodies for use in treating cancer in a patient, wherein the patient is refractory to or has relapsed after the anti-CD19 antibody treatment, and is treated with CD19-directed CAR-T cells, and wherein the anti-CD19 antibody comprises tafasitamab, and the CD19-directed CAR-T cells comprise one of axicabtagene ciloleucel (YESCARTA®) and / or tisagenlecleucel (KYMRIAH®), lysocabtagene maraleucel (JCAR017), or UCART19 (S68587), or a combination of two or more thereof.

[0079] In other embodiments, the CAR-T cells are one of the CAR-T cell constructs known as KITE037, welgenaleucel, ICTCAR-003, IM-19, CTX-110, SSCAR-010, ICTCAR-011, or a combination of two or more of these.

[0080] In another aspect, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and chimeric antigen receptor (CAR) T cells directed against CD19 for use in treating cancer.

[0081] In certain aspects, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient.

[0082] In another aspect, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient, and the anti-CD19 antibody is administered at least once every two weeks.

[0083] In a further aspect, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient, and the anti-CD19 antibody is administered in combination with one or more additional pharmaceutical agents.

[0084] In another aspect, a sequential therapeutic combination is disclosed for use in treating cancer, comprising an anti-CD19 antibody and CD19-directed CAR-T cells, wherein the anti-CD19 antibody is administered to a cancer patient at least once every two weeks, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient, and the anti-CD19 antibody is administered in combination with one or more additional pharmaceutical agents.

[0085] In one embodiment, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient, and the anti-CD19 antibody is administered in combination with one or more additional pharmaceutical agents, wherein the pharmaceutical agents are biologic or chemotherapeutic agents or pharmaceutically acceptable salts thereof.

[0086] In a further aspect, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient, and the anti-CD19 antibody is administered in combination with one or more additional pharmaceutical agents, wherein the pharmaceutical 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.

[0087] In a further aspect, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient, and the anti-CD19 antibody is administered in combination with one or more additional pharmaceutical agents, and the pharmaceutical agents are selected from the group of 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.

[0088] In a preferred embodiment, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient, and the anti-CD19 antibody is administered in combination with one or more additional pharmaceutical agents, wherein the pharmaceutical agent is lenalidomide or a pharmaceutically acceptable salt thereof.

[0089] In a particular aspect, a sequential therapeutic combination is disclosed for use in treating cancer, comprising an anti-CD19 antibody and CD19-directed CAR-T cells, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed CAR-T cells are administered to the patient, and the anti-CD19 antibody is administered in combination with lenalidomide or a pharmaceutically acceptable salt thereof, and the anti-CD19 antibody is administered at least once every two weeks in an amount of 12 mg / kg per dose, and lenalidomide is administered daily in an amount of 25 mg.

[0090] In certain aspects, a sequential therapeutic combination is disclosed comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences a relapse or if the patient is refractory to treatment, CD19-directed chimeric antigen receptor (CAR) T cells are administered to the patient, wherein the anti-CD19 antibody is administered in combination with lenalidomide or a pharmaceutically acceptable salt thereof, and wherein the anti-CD19 antibody is administered at least once every two weeks in an amount of 12 mg / kg per dose, and lenalidomide is administered daily in an amount of 25 mg, and wherein the anti-CD19 antibody is administered in a regimen of up to 12 cycles, wherein during cycles 1-3, the anti-CD19 antibody is administered weekly, and from cycle 4 onwards, the anti-CD19 antibody is administered every 14 days, and lenalidomide is administered daily.

[0091] In a preferred embodiment, in the sequential therapeutic combination disclosed herein comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, the anti-CD19 antibody has 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).

[0092] In other embodiments, in the sequential therapeutic combinations disclosed herein comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in the treatment of cancer, the anti-CD19 antibody has a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8. In particular embodiments, the sequential therapeutic combinations disclosed herein comprise an anti-CD19 antibody and CD19-directed CAR-T cells for use in the treatment of cancer, the anti-CD19 antibody having a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12. In some embodiments, the sequential therapeutic combinations disclosed herein comprise tafasitamab and CD19-directed CAR-T cells for use in the treatment of cancer.

[0093] In a preferred embodiment, in the sequential therapeutic combination disclosed herein comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in the treatment of cancer, the anti-CD19 antibody has i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab, and CD19-directed CAR-T cells include one or a combination of two or more of axicabtagene-ciloleucel (KTE-C19, Axi-cel, YESCARTA®) and / or tisagenlecleucel (CTL019, KYMRIAH®), lysocabtagene-malaleucel (JCAR017), or UCART19 (S68587). In another embodiment, the CAR-T cells are one or a combination of two or more of the CAR-T cell constructs known as KITE037, welgenaleucel, ICTCAR-003, IM-19, CTX-110, SSCAR-010, or ICTCAR-011.

[0094] In a preferred embodiment, in the sequential therapy combination disclosed herein comprising an anti-CD19 antibody and CD19-directed CAR-T cells for use in treating cancer, the cancer is a hematological cancer selected from non-Hodgkin's B-cell lymphomas, such as follicular lymphoma (FL), small lymphocytic lymphoma (SLL), Burkitt's lymphoma, and diffuse large B-cell lymphoma (DLBCL), Waldenstrom's hypergammaglobulinemia, leukemia, such as chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), or acute myeloid leukemia (AML). In a preferred embodiment, the hematological cancer is DLBCL. In another embodiment, the cancer is a relapsed or refractory (R / R) cancer, preferably a relapsed or refractory hematological cancer. The R / R hematological cancer is selected from R / R non-Hodgkin's B-cell lymphomas, such as R / R follicular lymphoma (FL), R / R small lymphocytic lymphoma (SLL), R / R Burkitt's lymphoma, and R / R diffuse large B-cell lymphoma (DLBCL), R / R Waldenstrom's hypergammaglobulinemia, R / R leukemias, such as R / R chronic lymphocytic leukemia (CLL), R / R acute lymphoblastic leukemia (ALL), or R / R acute myeloid leukemia (AML). In a preferred embodiment, the R / R hematological cancer is relapsed or refractory (R / R) diffuse large B-cell lymphoma (DLBCL).

[0095] In one preferred embodiment, the sequential therapeutic combination comprises tafasitamab and axicabtagene-ciloleucel (YESCARTA®) CAR-T cells for use in the treatment of DLBCL in patients who are refractory or have relapsed to prior treatment.

[0096] In one preferred embodiment, the sequential therapeutic combination comprises tafasitamab and axicabtagene-ciloleucel (YESCARTA®) CAR-T cells for use in treating DLBCL in a patient who is refractory to or has relapsed from a prior treatment, wherein the prior treatment comprises tafasitamab.

[0097] In another embodiment, an anti-CD19 antibody is disclosed for use in treating cancer in a patient, wherein the patient has previously been treated with a composition comprising a therapeutic agent directed against CD19. In one particular embodiment, an anti-CD19 antibody is disclosed for use in treating cancer in a patient, wherein the patient has previously been treated with a composition comprising CAR-T cells directed against CD19.

[0098] In a preferred embodiment, an anti-CD19 antibody is disclosed having 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), for use in treating cancer in a patient, wherein the patient is refractory to or has relapsed from a prior treatment, and the prior treatment comprises CAR-T cells directed against CD19.

[0099] In another embodiment, an anti-CD19 antibody having a variable heavy chain comprising SEQ ID NO:7 and / or a variable light chain comprising SEQ ID NO:8 is disclosed for use in treating cancer in a patient, wherein the patient is refractory to or has experienced a relapse after a prior treatment, wherein the prior treatment comprised CAR-T cells directed against CD19.

[0100] In one embodiment, an anti-CD19 antibody having a variable heavy chain and / or variable light chain with at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the variable heavy chain of SEQ ID NO: 7 and / or the variable light chain of SEQ ID NO: 8 is disclosed for use in treating cancer in a patient, wherein the patient is refractory to or has relapsed from a prior treatment, wherein the prior treatment included CAR-T cells directed against CD19.

[0101] In certain embodiments, an anti-CD19 antibody having a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12 is disclosed for use in treating cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising CAR-T cells directed against CD19.

[0102] In certain embodiments, anti-CD19 antibodies, including tafasitamab, are disclosed for use in treating cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising CAR-T cells directed against CD19.

[0103] In another embodiment, an anti-CD19 antibody is disclosed for use in treating cancer in a patient who is refractory or has relapsed from a prior treatment, the prior treatment comprising CD19-directed CAR-T cells in which a chimeric antigen receptor competes with the anti-CD19 antibody for binding to human CD19.

[0104] In other embodiments, an anti-CD19 antibody is disclosed for use in treating cancer in a patient, wherein the patient is refractory to or has experienced a relapse after a prior treatment, wherein the prior treatment comprises an anti-CD19 therapeutic agent, and wherein the therapeutic agent is selected from the group of an antibody, an antibody-drug conjugate, a bispecific antibody, an alternative scaffold protein, or a CAR-T cell.

[0105] In a preferred embodiment, an anti-CD19 antibody is disclosed for use in treating cancer in a patient who is refractory to or has relapsed from a prior treatment, the prior treatment comprising anti-CD19 CAR-T cells, and the anti-CD19 antibody comprises tafasitamab. In certain embodiments, the CAR-T cells are preferably one of axicabtagene-ciloleucel (KTE-C19, Axi-cel, YESCARTA®) and / or tisagenlecleucel (CTL019, KYMRIAH®), lisocabtagene-malaleucel (JCAR017, clinical trials: e.g., NCT02631044, NCT03310619), or UCART19, or a combination of two or more thereof. In another embodiment, the CAR-T cells are one of the CAR-T cell constructs known as KITE037, welgenaleucel, ICTCAR-003, IM-19, CTX-110, SSCAR-010, ICTCAR-011, or a combination of two or more thereof.

[0106] In other embodiments, the antibody is directed against a B-cell lineage marker for use in treating cancer in a patient, wherein the patient is refractory to or has experienced a relapse after a prior treatment, the prior treatment comprising a therapeutic anti-CD19 agent.

[0107] In another embodiment, the antibody is directed against CD19 and / or CD20 for use in treating cancer in a patient who is refractory or has experienced a relapse to prior treatment, the prior treatment comprising a therapeutic anti-CD19 agent.

[0108] In other embodiments, the antibody is directed against a B-cell lineage marker for use in treating cancer in a patient who is refractory or has experienced a relapse to prior treatment, the prior treatment comprising CAR-T cells directed against CD19.

[0109] In another embodiment, the antibody is directed against CD19 and / or CD20 for use in treating cancer in a patient, wherein the patient is refractory to or has relapsed from a prior treatment, the prior treatment comprising CAR-T cells directed against CD19.

[0110] In preferred embodiments, the antibodies directed against B cell lineage markers (e.g., CD19 and / or CD20) are for use in pharmaceutical compositions. In other embodiments, the antibodies directed against B cell lineage markers (e.g., CD19 and / or CD20) are included in pharmaceutical compositions.

[0111] In one embodiment, an anti-CD19 antibody for use in treating cancer in a patient is disclosed, wherein the patient has previously been treated with a composition comprising CD19-directed CAR-T cells, and the cancer is a hematological cancer. In certain embodiments, the hematological cancer is selected from non-Hodgkin's B-cell lymphomas, such as follicular lymphoma (FL), small lymphocytic lymphoma (SLL), Burkitt's lymphoma, and diffuse large B-cell lymphoma (DLBCL), Waldenstrom's hypergammaglobulinemia, and leukemias, such as chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), or acute myeloid leukemia (AML). In a preferred embodiment, the hematological cancer is DLBCL. In other embodiments, the cancer is a relapsed or refractory (R / R) cancer, preferably a relapsed or refractory hematological cancer. The R / R hematological cancer is selected from R / R non-Hodgkin's B-cell lymphomas, such as R / R follicular lymphoma (FL), R / R small lymphocytic lymphoma (SLL), R / R Burkitt's lymphoma, and R / R diffuse large B-cell lymphoma (DLBCL), R / R Waldenstrom's hypergammaglobulinemia, R / R leukemias, such as R / R chronic lymphocytic leukemia (CLL), R / R acute lymphoblastic leukemia (ALL), or R / R acute myeloid leukemia (AML). In a preferred embodiment, the R / R hematological cancer is relapsed or refractory (R / R) diffuse large B-cell lymphoma (DLBCL).

[0112] In certain embodiments, an anti-CD19 antibody is disclosed for use in treating R / R DLBCL in a patient who is refractory or has experienced a relapse to a prior treatment, the prior treatment comprising CAR-T cells directed against CD19, the anti-CD19 antibody having i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0113] In certain embodiments, a combination of tafasitamab and lenalidomide is disclosed for use in treating R / R DLBCL in a patient who is refractory or has relapsed to a prior treatment, wherein the prior treatment comprises CAR-T cells directed against CD19.

[0114] In another embodiment, a combination of tafasitamab and bendamustine is disclosed for use in treating R / R DLBCL in a patient who is refractory or has relapsed to a prior treatment, wherein the prior treatment comprises CAR-T cells directed against CD19.

[0115] treatment method In one embodiment, a method of treating cancer with CD19-directed CAR-T cells in a patient is disclosed, wherein the patient is refractory to or has experienced a relapse after a prior treatment, the prior treatment comprising an anti-CD19 antibody.

[0116] In one embodiment, a method of treating cancer with a pharmaceutical composition comprising CD19-directed CAR-T cells in a patient is disclosed, wherein the patient is refractory to or has experienced a relapse following a prior treatment, the prior treatment comprising an anti-CD19 antibody.

[0117] In one embodiment, a method is disclosed for treating cancer with a pharmaceutical composition comprising CAR-T cells directed against CD19 in a patient in a second, third, fourth, or any other multiple treatment regimen, where the patient is refractory or has experienced a relapse to a prior treatment regimen, and the prior regimen comprises an anti-CD19 antibody.

[0118] In another embodiment, a method is disclosed for treating cancer with a pharmaceutical composition comprising CAR-T cells directed against CD19 in a patient in a second, third, fourth, or any other multiple treatment regimen, wherein the patient received a pharmaceutical composition comprising an anti-CD19 antibody in at least one of the prior treatment regimen(s).

[0119] In another embodiment, a method is disclosed for treating cancer with a pharmaceutical composition comprising CAR-T cells directed against CD19 in a patient in a second, third, fourth, or any other multiple treatment regimen, wherein the patient has been administered a pharmaceutical composition comprising an anti-CD19 antibody in at least one of the prior treatment regimen(s), and the patient is refractory or has experienced a relapse after at least one of the prior treatment regimen(s).

[0120] In another embodiment, a method is disclosed for treating hematological cancer with a pharmaceutical composition comprising CAR-T cells directed against CD19 in a patient in a second, third, fourth, or any other multiple treatment regimen, wherein the patient received a pharmaceutical composition comprising tafasitamab in at least one of the prior treatment regimen(s).

[0121] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising an anti-CD19 antibody, the antibody having 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); and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising a CD19-directed therapeutic agent.

[0122] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising an anti-CD19 antibody, the antibody having a variable heavy chain of SEQ ID NO: 7 and / or a variable light chain of SEQ ID NO: 8; and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising a CD19-directed therapeutic agent.

[0123] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising an anti-CD19 antibody, the antibody having a heavy chain of SEQ ID NO: 11 and / or a light chain of SEQ ID NO: 12; and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising a CD19-directed therapeutic agent.

[0124] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying patients who have received a pharmaceutical composition comprising the anti-CD19 antibody tafasitamab; and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising a CD19-directed therapeutic agent.

[0125] In certain embodiments, the CD19-directed therapeutic agent of the second, third, fourth, or any other multiple therapeutic regimen in step (ii) is a monoclonal antibody, an antibody-drug conjugate, a bispecific antibody, an alternative scaffold protein, or a chimeric antigen receptor (CAR) T cell. Preferably, the CD19-directed therapeutic agent of the pharmaceutical composition of the second, third, fourth, or any other multiple therapeutic regimen is a CAR-T cell.

[0126] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying patients who have received a pharmaceutical composition comprising the anti-CD19 antibody tafasitamab; and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising CAR-T cells directed against CD19.

[0127] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying patients who have received a pharmaceutical composition comprising the anti-CD19 antibody tafasitamab; and ii) administering to the patient in the second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising CAR-T cells directed against CD19, the CAR-T cells being selected from one of axicabtagene ciloleucel, tisagenlecleucel, lysocabtagene maraleucel, and / or UCART19, or a combination thereof.

[0128] In one embodiment, a method of treating R / R DLBCL in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying patients who have received a pharmaceutical composition comprising the anti-CD19 antibody tafasitamab; and ii) administering to the patient in the second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising CAR-T cells directed against CD19, the CAR-T cells being selected from one of axicabtagene ciloleucel, tisagenlecleucel, lysocabtagene maraleucel, and / or UCART19, or a combination thereof.

[0129] In some embodiments, a method of treating a relapsed or refractory CD19+ hematological cancer with a therapeutic anti-CD19 agent is disclosed, the method comprising the steps of: a) identifying patients with relapsed or refractory CD19+ hematologic cancers; and b) administering to said patient an effective amount of a CD19-directed therapeutic agent; The patient has been treated prior to step a) with a pharmaceutical composition comprising an anti-CD19 antibody, the antibody having: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0130] In certain embodiments, a method of treating relapsed or refractory CD19+ hematological cancer using CD19-directed CAR-T cells is disclosed, the method comprising the steps of: a) identifying patients with relapsed or refractory CD19+ hematologic cancers; and b) administering to said patient an effective amount of CD19-directed CAR-T cells; The patient has been treated prior to step a) with a pharmaceutical composition comprising an anti-CD19 antibody, the antibody having: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0131] In certain embodiments, a method for treating relapsed or refractory DLBCL using CD19-directed CAR-T cells is disclosed, the method comprising the steps of: a) to identify patients with relapsed or refractory DLBCL, and b) administering to said patient an effective amount of CD19-directed CAR-T cells; The patient has been treated prior to step a) with a pharmaceutical composition comprising an anti-CD19 antibody, the antibody having: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0132] In some embodiments, provided are methods of treatment that involve: (1) administering to a patient having cancer a composition comprising an anti-CD19 antibody, the antibody having: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab, and (2) if the patient is relapsed or refractory, sequentially administering to the patient a composition comprising T cells that are autologous to the patient and express a recombinant receptor that specifically binds to CD19.

[0133] In a preferred embodiment, the CD19-directed autologous T cells for use in treating cancer in a patient are chimeric antigen receptor (CAR) T cells, provided the patient has previously been treated with a composition comprising an anti-CD19 antibody. In certain embodiments, the CAR T cells are preferably one of axicabtagene-ciloleucel (KTE-C19, Axi-cel, YESCARTA®) and / or tisagenlecleucel (CTL019, KYMRIAH®), lisocabtagene-malaleucel (JCAR017, clinical trials: e.g., NCT02631044, NCT03310619), or UCART19, or a combination of two or more thereof. In another embodiment, the CAR-T cells are one of the CAR-T cell constructs known as KITE037, welgenaleucel, ICTCAR-003, IM-19, CTX-110, SSCAR-010, ICTCAR-011, or a combination of two or more thereof.

[0134] In one embodiment, a method of treating cancer with an anti-CD19 antibody in a patient is disclosed, wherein the patient is refractory or has experienced a relapse to a prior treatment, the prior treatment comprising CAR-T cells directed against CD19.

[0135] In one embodiment, a method of treating cancer in a patient with a composition comprising an anti-CD19 antibody is disclosed, wherein the patient is refractory to or has experienced a relapse after a prior treatment, the prior treatment comprising CAR-T cells directed against CD19.

[0136] In one embodiment, a method is disclosed for treating cancer in a patient in a second, third, fourth, or any other multiple treatment regimen with a pharmaceutical composition comprising an anti-CD19 antibody, where the patient is refractory or has experienced a relapse and has received a pharmaceutical composition comprising CAR-T cells directed against CD19 in a prior treatment regimen.

[0137] In another embodiment, a method is disclosed for treating cancer in a second, third, fourth, or any other multiple treatment regimen with a pharmaceutical composition comprising an anti-CD19 antibody in a patient who is refractory or has experienced a relapse and who has received a pharmaceutical composition comprising CAR-T cells directed against CD19 in one of the prior treatment regimen(s).

[0138] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising CD19-directed CAR-T cells; and ii) in the second, third, fourth, or any other multiple treatment regimen, administering to the patient a further pharmaceutical composition comprising an anti-CD19 antibody, the antibody having 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).

[0139] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising CD19-directed CAR-T cells; and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising an anti-CD19 antibody, wherein the antibody has a variable heavy chain of SEQ ID NO:7 and / or a variable light chain of SEQ ID NO:8.

[0140] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising CD19-directed CAR-T cells; and ii) administering to the patient in the second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising an anti-CD19 antibody, wherein the antibody has a heavy chain of SEQ ID NO: 11 and / or a light chain of SEQ ID NO: 12.

[0141] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising CD19-directed CAR-T cells; and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising the anti-CD19 antibody tafasitamab.

[0142] In one embodiment, a method of treating cancer in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising CAR-T cells directed against CD19, the CAR-T cells being selected from one or a combination of axicabtagene ciloleucel, tisagenlecleucel, lysocabtagene maraleucel, and / or UCART19; and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising the anti-CD19 antibody tafasitamab.

[0143] In one embodiment, a method of treating R / R DLBCL in a patient with a second, third, fourth, or any other multiple treatment regimen is disclosed, the method comprising the steps of: i) identifying a patient who has been administered a pharmaceutical composition comprising CAR-T cells directed against CD19, the CAR-T cells being selected from one or a combination of axicabtagene ciloleucel, tisagenlecleucel, lysocabtagene maraleucel, and / or UCART19; and ii) administering to the patient in said second, third, fourth, or any other multiple treatment regimen an additional pharmaceutical composition comprising the anti-CD19 antibody tafasitamab.

[0144] In some embodiments, a method of treating a relapsed or refractory CD19+ hematological cancer with an anti-CD19 antibody is disclosed, the method comprising the steps of: a) identifying patients with relapsed or refractory CD19+ hematologic cancers; and b) administering to the patient an effective amount of an anti-CD29 antibody, wherein the patient has been treated prior to step (a) with a pharmaceutical composition comprising CAR-T cells directed against CD19, and the anti-CD19 antibody of step (b) has i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0145] In some aspects, provided are treatments that involve: (1) administering to a patient with cancer a composition comprising T cells that express a recombinant receptor that specifically binds to CD19 that is autologous to the patient and associated with the cancer; and (2) if the patient is refractory to treatment or experiences a relapse, sequentially administering to the patient a composition comprising an anti-CD19 antibody, the antibody comprising: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0146] In a preferred embodiment, the CD19-directed autologous T cells for use in treating cancer in a patient are chimeric antigen receptor (CAR) T cells when the patient is refractory or experiencing relapse, and when the patient is sequentially treated with a composition comprising an anti-CD19 antibody. In certain embodiments, the CAR T cells are preferably one of axicabtagene-ciloleucel (KTE-C19, Axi-cel, YESCARTA®) and / or tisagenlecleucel (CTL019, KYMRIAH®), lisocabtagene-malaleucel (JCAR017, clinical trials: e.g., NCT02631044, NCT03310619), or UCART19, or a combination of two or more thereof. In another embodiment, the CAR-T cells are one of the (CAR) T-cell constructs known as KITE037, welgenaleucel, ICTCAR-003, IM-19, CTX-110, SSCAR-010, ICTCAR-011, or a combination of two or more of these.

[0147] How to predict the response In another embodiment, a method for determining whether a patient is likely or unlikely to respond to CD19-directed CAR-T cell therapy following treatment with tafasitamab is disclosed, the method comprising the steps of: a) contacting a tumor sample isolated from a patient with an effective amount of an anti-CD19 antibody having a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8 to saturate the antigen in vitro; b) adding CD19-directed CAR-T cells at various effector:target ratios; and c) assessing CAR-T cell effector function (e.g., antigen-specific killing, degranulation, cytokine production, or proliferation); In this case, the presence of CAR-T cell effector function indicates that this patient is likely to respond to anti-CD19 CAR-T cell therapy, and the absence of CAR-T cell effector function indicates that this patient is unlikely to respond to anti-CD19 CAR-T cell therapy.

[0148] In another embodiment, a method is disclosed for determining whether a treatment-resistant or relapsed patient is likely or unlikely to respond to CD19-directed CAR-T cell therapy following treatment with tafasitamab, the method comprising the steps of: a) contacting a tumor sample isolated from a patient with an effective amount of an anti-CD19 antibody having a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8 to saturate the antigen in vitro; b) adding CD19-directed CAR-T cells at various effector:target ratios; and c) assessing CAR-T cell effector function (e.g., antigen-specific killing, degranulation, cytokine production, or proliferation); In this case, the presence of CAR-T cell effector function indicates that this patient is likely to respond to anti-CD19 CAR-T cell therapy, and the absence of CAR-T cell effector function indicates that this patient is unlikely to respond to anti-CD19 CAR-T cell therapy.

[0149] In specific embodiments, the various effector:target (E:T) ratios are 0:1, 0.3125:1, 0.625:1, 1.25:1, 2.5:1, 5:1, and 10:1. In one embodiment, the anti-CD19 CAR-T cells comprise a CAR construct of the format FMC63-CD8h-CD8TM-41BBζ.

[0150] 1. Use of a CD19-directed therapeutic agent in the manufacture of a medicament for treating a hematological cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising an anti-CD19 antibody, the antibody having: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0151] 1. Use of CD19-directed CAR-T cells in the manufacture of a medicament for treating a hematological cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising an anti-CD19 antibody, the antibody comprising: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0152] 1. Use of CD19-directed CAR-T cells in the manufacture of a medicament for treating a hematological cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising an anti-CD19 antibody, the antibody comprising: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab, and the CD19-directed CAR-T cells are selected from the group consisting of axicabtagene-ciloleucel (YESCARTA®), tisagenlecleucel (KYMRIAH®), lysocabtagene-malaleucel, or UCART19.

[0153] 1. Use of CD19-directed CAR-T cells in the manufacture of a medicament for treating a hematological cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising an anti-CD19 antibody, the antibody comprising: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab, and the hematological cancers include non-Hodgkin's B-cell lymphomas, such as follicular lymphoma (FL), small lymphocytic lymphoma (SLL), Burkitt's lymphoma, and diffuse large B-cell lymphoma (DLBCL), Waldenstrom's hypergammaglobulinemia, leukemias, such as chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), or acute myeloid leukemia (AML), relapsed or refractory (R / R) non-Hodgkin's B-cell lymphomas. The target lymphoma is selected from lymphomas such as R / R follicular lymphoma (FL), R / R small lymphocytic lymphoma (SLL), R / R Burkitt's lymphoma, and R / R diffuse large B-cell lymphoma (DLBCL), R / R Waldenstrom's hypergammaglobulinemia, and R / R leukemias such as R / R chronic lymphocytic leukemia (CLL), R / R acute lymphoblastic leukemia (ALL), and R / R acute myeloid leukemia (AML), preferably R / R DLBCL.

[0154] Use of CD19-directed axicabtagene-ciloleucel (YESCARTA®) CAR-T cells in the manufacture of a medicament for treating R / R DLBCL in a patient who is refractory to or has relapsed from prior treatment, the prior treatment comprising the anti-CD19 antibody tafasitamab.

[0155] 1. Use of an anti-CD19 antibody in the manufacture of a medicament for treating a hematological cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising CAR-T cells directed against CD19, and the anti-CD19 antibody having: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0156] 1. Use of an anti-CD19 antibody in the manufacture of a medicament for treating a hematological cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising CAR-T cells directed against CD19, and the anti-CD19 antibody comprising: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab, and the CD19-directed CAR-T cells are selected from the group consisting of axicabtagene-ciloleucel (YESCARTA®), tisagenlecleucel (KYMRIAH®), lysocabtagene-malaleucel, or UCART19.

[0157] 1. Use of an anti-CD19 antibody in the manufacture of a medicament for treating a hematological cancer in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising CAR-T cells directed against CD19, and the antibody comprising: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab, and the hematological cancers include non-Hodgkin's B-cell lymphomas, such as follicular lymphoma (FL), small lymphocytic lymphoma (SLL), Burkitt's lymphoma, and diffuse large B-cell lymphoma (DLBCL), Waldenstrom's hypergammaglobulinemia, leukemias, such as chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), or acute myeloid leukemia (AML), relapsed or refractory (R / R) non-Hodgkin's B-cell lymphomas. The target lymphoma is selected from lymphomas such as R / R follicular lymphoma (FL), R / R small lymphocytic lymphoma (SLL), R / R Burkitt's lymphoma, and R / R diffuse large B-cell lymphoma (DLBCL), R / R Waldenstrom's hypergammaglobulinemia, and R / R leukemias such as R / R chronic lymphocytic leukemia (CLL), R / R acute lymphoblastic leukemia (ALL), and R / R acute myeloid leukemia (AML), preferably R / R DLBCL.

[0158] 1. Use of an anti-CD19 antibody in the manufacture of a medicament for treating R / R DLBCL in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising CAR-T cells directed against CD19, and the antibody having: i) 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), or ii) a variable heavy chain of SEQ ID NO: 7 and a variable light chain of SEQ ID NO: 8, or iii) a heavy chain of SEQ ID NO: 11 and a light chain of SEQ ID NO: 12, or iv) tafasitamab.

[0159] Use of tafasitamab in the manufacture of a medicament for treating R / R DLBCL in a patient, the patient being refractory to or experiencing a relapse after a prior treatment, the prior treatment comprising CAR-T cells directed against CD19.

Claims

1. 1. A CD19-directed therapeutic agent for use in treating cancer in a patient, wherein the patient is resistant to or has experienced a relapse after a prior treatment, the prior treatment comprising an anti-CD19 antibody having 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).

2. 10. The CD19-directed therapeutic for use according to any one of the preceding claims, wherein said therapeutic is selected from the group of an antibody, an antibody drug conjugate, a bispecific antibody, an alternative scaffold protein, or a chimeric antigen receptor (CAR) T cell.

3. 3. The CD19-directed therapeutic for use according to claim 2, wherein the therapeutic is a chimeric antigen receptor (CAR) T cell.

4. The CD19-directed chimeric antigen receptor (CAR) T cells for use according to claim 3, wherein the CAR-T cells are axicabtagene ciloleucel, tisagenlecleucel, lysocabtagene maraleucel, or UCART19.

5. 10. The CD19 directed therapeutic agent for use according to any one of the preceding claims, wherein the cancer is a hematological cancer and is selected from non-Hodgkin's B-cell lymphoma, follicular lymphoma (FL), small lymphocytic lymphoma (SLL), Burkitt's lymphoma, diffuse large B-cell lymphoma (DLBCL), Waldenstrom's hypergammaglobulinemia, B-cell leukemia, chronic lymphocytic leukemia (CLL), or acute lymphoblastic leukemia (ALL).

6. The CD19-directed therapeutic agent for use according to claim 5, wherein the hematological cancer is a relapsed or refractory hematological cancer.

7. The CD19-directed therapeutic agent for use according to claim 6, wherein the relapsed or refractory hematological cancer is R / R-DLBCL.

8. 1. An anti-CD19 antibody for use in treating cancer in a patient, wherein the patient has been treated with chimeric antigen receptor (CAR) T cells directed against CD19 after the anti-CD19 antibody treatment, and the anti-CD19 antibody has 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).

9. 1. A sequential therapeutic combination comprising an anti-CD19 antibody and chimeric antigen receptor (CAR) T cells directed against CD19 for use in the treatment of cancer.

10. 10. The sequential therapeutic combination of claim 9 for use in treating cancer, wherein the anti-CD19 antibody is administered to a cancer patient, and after the patient experiences relapse or if the patient is refractory to treatment, CAR-T cells directed against CD19 are administered to the patient.

11. 11. The sequential therapeutic combination of claim 10 for use in the treatment of cancer, wherein the anti-CD19 antibody is administered in combination with one or more additional pharmaceutical agents.

12. 12. The sequential therapeutic combination of claim 11 for use in the treatment of cancer, wherein the pharmaceutical agent is a therapeutic antibody or antibody fragment, nitrogen mustard, purine analog, thalidomide analog, phosphoinositide 3-kinase inhibitor, BCL-2 inhibitor, Bruton's tyrosine kinase (BTK) inhibitor, or a pharmaceutically acceptable salt thereof.

13. 13. The sequential therapeutic combination of claim 12 for use in the treatment of cancer, wherein the pharmaceutical agent is lenalidomide or a pharmaceutically acceptable salt thereof.

14. 14. The sequential therapeutic combination of claim 13 for use in the treatment of cancer, wherein the anti-CD19 antibody is administered in a regimen of up to 12 cycles, wherein during cycles 1-3, the anti-CD19 antibody is administered weekly, and from cycle 4 onwards, the anti-CD19 antibody is administered every 14 days, and lenalidomide is administered daily.

15. 15. The sequential therapeutic combination of any one of claims 9 to 14, wherein the anti-CD19 antibody has 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).

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