Dosage regimen and use of anti-CD19 agents

Abstract

The present disclosure relates to dosing regimens of anti-CD19 agents, particularly anti-CD19×anti-CD3×anti-CD2 trispecific agents, administered intravenously (iv) and subcutaneously (sc), and their use for treating diseases and disorders associated with expression of CD19, such as B cell malignancies, particularly relapsed and / or refractory B cell malignancies.

Description

[Technical field]

[0001] The present disclosure relates generally to intravenously (iv) and subcutaneously (sc) administered anti-CD19 agents, in particular anti-CD19×anti-CD3×anti-CD2 trispecific agents, and their use to treat diseases and disorders associated with expression of CD19, such as B cell malignancies, in particular relapsed and / or refractory B cell malignancies. [Background technology]

[0002] B cells express a wide range of cell surface molecules during their differentiation and proliferation. CD19 is a pan-B cell membrane glycoprotein expressed from early stages of pre-B cell development through terminal differentiation and regulates B lymphocyte development and function. CD19 expression has been identified in the majority of non-Hodgkin's lymphomas (NHL) and leukemias, including chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), and Waldenstrom's macroglobulinemia (WM).

[0003] Patients with relapsed and / or refractory (R / R) NHL who have failed treatment after initial therapy have poor outcomes. Approximately 50% of patients have a response to initial salvage therapy and then undergo allogeneic stem cell transplantation (ASCT), with an overall cure rate in the range of 25 to 35%. Patients who are not candidates for ASCT due to poor fitness due to age or coexisting medical conditions, patients who have no response to salvage therapy, and patients who have relapse after ASCT are classified as transplant-ineligible. Finally, most patients with R / R NHL belong to this category, and sequential single-agent chemotherapy or multi-agent regimens with acceptable side effect profiles, such as rituximab, gemcitabine, and oxaliplatin (R-GemOx), are frequently used with palliative intent.

[0004] Despite advances in the treatment of R / R ALL, disease recurrence and relapse in patients with increasingly refractory disease remain major obstacles. Treatment options for R / R ALL patients remain limited and include high-dose chemotherapy followed by stem cell transplantation (SCT), standard chemoimmunotherapy, CD19- or CD22-directed therapy (blinatumomab, inotuzumab ozogamicin), targeted therapy with small molecule pathway inhibitors, or non-curative palliative supportive care, but the duration of remission is typically short.

[0005] NHLs are sometimes classified into immature lymphoid neoplasms, mature B-cell neoplasms, T-cell and natural killer (NK) cell neoplasms, and post-transplant lymphoproliferative disorders (PTLDs). Mature B-cell lymphomas can be further classified into indolent lymphomas (e.g., follicular lymphoma, small lymphocytic lymphoma, marginal zone lymphoma) and aggressive lymphomas (e.g., diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL)).

[0006] DLBCL is the most frequent aggressive lymphoma subtype, representing 30–35% of all NHL (Ghielmini et al., 2013). Approximately one-third of DLBCL patients will develop relapsed and / or refractory (R / R) disease, representing the main cause of morbidity and mortality. Relapsed and refractory patients have a poor prognosis. In the SCHOLAR-1 study, which combined data from two clinical trials and two academic databases, the median overall survival of 636 patients with refractory DLBCL was only 6.3 months, while only 20% of patients were alive after 2 years (Crump et al., 2017).

[0007] Follicular lymphoma (FL) is the second most common type of NHL. It is the most common clinically indolent NHL, defined as a lymphoma in which survival of untreated patients is measured over a period of years. The majority of patients treated for FL have an initial response to treatment, with 40-80% experiencing complete remission depending on the initial regimen used. However, conventional therapies for FL are not curative, and the majority of these patients will eventually develop progressive disease. There remains a high unmet medical need for patients with R / R FL.

[0008] Acute lymphoblastic lymphoma (ALL) is characterized by the accumulation of malignant lymphoblasts in the bone marrow, peripheral circulation, and lymphoid organs. Blinatumomab, a CD19-CD3 bispecific T-cell engager, has been approved for the treatment of ALL. However, treatment with blinatumomab lacks durable responses and is characterized by a high relapse rate. Von Stackelberg et al., 2016, Journal of Clinical Oncology 34(36):4381-4389. Furthermore, blinatumomab has a short half-life, requiring continuous exposure for the drug to exert sufficient efficacy and manageable toxicity. Porter et al., 2013, Clin Pharmacol. 5(Suppl1):5-11. Summary of the Invention

[0009] Despite major improvements in cancer therapy, B-cell malignancies, such as the B-cell subtype of non-Hodgkin's lymphoma, contribute primarily to cancer-related deaths. In particular, patients with R / R B-NHL and R / R B-ALL who have failed multiple existing first-line therapies have a high unmet medical need. There remains a high unmet medical need for patients with R / R DLBCL who are not eligible for or do not have access to autologous stem cell transplantation (ASCT) and CAR-T therapy. These patients have few effective treatment options and have significantly reduced overall survival (OS). There is also a high unmet need for patients who relapse after transplantation or CAR-T therapy. Thus, there remains a need for additional therapeutic agents to treat certain B-cell malignancies.

[0010] In some embodiments, the present invention relates to a method of treating a subject having a B cell malignancy by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in treating a B cell malignancy. The B cell malignancy may be a relapsed and / or refractory B cell malignancy. The B cell malignancy may be selected from the group consisting of R / R LBCL, R / R DLBCL, R / R HGBCL, R / R PMBCL, R / R FL, R / R FL3B, R / R MCL, R / R SLL, R / R MZL and R / R ALL.

[0011] In some embodiments, the present invention relates to a method of treating a subject having a CD19-related disease or disorder by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in treating a CD19-related disease or disorder. The CD19-related disease or disorder may be a relapsed and / or refractory disease or disorder. The CD19-related disease or disorder may be selected from the group consisting of R / R LBCL, R / R DLBCL, R / R HGBCL, R / R PMBCL, R / R FL, R / R FL3B, R / R MCL, R / R SLL, R / R MZL and R / R ALL. In some embodiments, the CD19-related disease or disorder is systemic lupus erythematosus (SLE).

[0012] In some embodiments, an anti-CD19xanti-CD3xanti-CD2 trispecific agent may comprise a CD19 binding moiety having CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, and CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences of SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.

[0013] In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent comprises the amino acid sequences given in Table 5.

[0014] In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise (a) a first polypeptide encoded by a nucleotide sequence comprising the sequence of SEQ ID NO: 40; (b) a second polypeptide encoded by a nucleotide sequence comprising the sequence of SEQ ID NO: 41; and (c) a third polypeptide encoded by a nucleotide sequence comprising the sequence of SEQ ID NO: 42. In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent is encoded by any of the nucleotide sequences given in Table 6.

[0015] In some embodiments, the anti-CD19×anti-CD3×anti-CD2 trispecific agent comprises any of the sequences provided in Tables 1 to 6. In some embodiments, the anti-CD19×anti-CD3×anti-CD2 trispecific agent is CD19TSP1.

[0016] In some embodiments, the invention relates to a method of treating a subject having a B cell malignancy by administering a trispecific antibody. In some embodiments, the invention relates to a trispecific antibody for use in treating a B cell malignancy. The B cell malignancy may be a relapsed and / or refractory B cell malignancy. In some embodiments, the trispecific antibody may comprise: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the trispecific antibody comprises the amino acid sequence given in Table 5. The B cell malignancy may be selected from the group consisting of LBCL, DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL, and ALL. The B-cell malignancy may be selected from the group consisting of R / R LBCL, R / R DLBCL, R / R HGBCL, R / R PMBCL, R / R FL, R / R FL3B, R / R MCL, R / R SLL, R / R MZL and R / R ALL.

[0017] In some embodiments, the invention relates to a method of treating a subject having a condition selected from LBCL, DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL and ALL, comprising administering to the subject a therapeutically effective amount of an anti-CD19×anti-CD3×anti-CD2 trispecific agent. In some embodiments, the invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of a condition selected from LBCL, DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL and ALL. In some embodiments, the invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of a condition selected from LBCL, DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL, ALL and SLE. In some embodiments, the condition is LBCL or FL3B, optionally relapsed and / or refractory LBCL or FL3B. In some embodiments, the condition is an autoimmune condition, optionally systemic lupus erythematosus (SLE).

[0018] In some embodiments, the present invention relates to a method of treating a subject having non-Hodgkin's lymphoma (NHL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The NHL may be relapsed and / or refractory NHL. The NHL may be relapsed and / or refractory B-cell non-Hodgkin's lymphoma (R / R B-NHL). In some embodiments, the present invention relates to a method of treating a subject having large B-cell lymphoma (LBCL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The LBCL may be relapsed and / or refractory LBCL. In some embodiments, the present invention relates to a method of treating a subject having diffuse large B-cell lymphoma (DLBCL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The DLBCL may be relapsed and / or refractory DLBCL. The DLBCL may be de novo or transformed. In some embodiments, the present invention relates to a method of treating a subject having high grade B cell lymphoma by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The lymphoma may be a double / triple hit high grade B cell lymphoma (HGBCL). The HGBCL may be a relapsed and / or refractory HGBCL. The HGBCL may be a relapsed and / or refractory double / triple hit HGBCL. In some embodiments, the present invention relates to a method of treating a subject having primary mediastinal large B cell lymphoma (PMBCL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The PMBCL may be a relapsed and / or refractory PMBCL. In some embodiments, the present invention relates to a method of treating a subject having follicular lymphoma (FL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The FL may be a relapsed and / or refractory FL.In some embodiments, the present invention relates to a method of treating a subject with follicular lymphoma grade 3B (FL3B) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The FL3B may be relapsed and / or refractory FL3B. In some embodiments, the present invention relates to a method of treating a subject with mantle cell lymphoma (MCL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The MCL may be relapsed and / or refractory MCL. In some embodiments, the present invention relates to a method of treating a subject with small lymphocytic lymphoma (SLL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The SLL may be relapsed and / or refractory SLL. In some embodiments, the present invention relates to a method of treating a subject with marginal zone lymphoma (MZL) (e.g., nodal, extranodal, or mucosal-associated) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The MZL may be relapsed and / or refractory MZL. Optionally, in each of the above cases, the treatment may be after a previous CAR-T therapy, e.g., a CD19-directed CAR-T therapy. In an alternative option, the treatment may be in the absence of a previous CAR-T therapy, e.g., a CD19-directed CAR-T therapy. Optionally, in each of the above cases, the treatment may be after treatment with a chemotherapy regimen comprising a CD20 monoclonal antibody. Further, optionally, in each of the above cases, the treatment may be after a previous autologous hematopoietic stem cell transplant (HSCT). The subject may have an Eastern Cooperative Oncology Group (ECOG) activity status of 2 or less.

[0019] In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of non-Hodgkin's lymphoma (NHL). The NHL lymphoma may be relapsed and / or refractory NHL. The NHL may be relapsed and / or refractory B-cell non-Hodgkin's lymphoma (R / R B-NHL). In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of large B-cell lymphoma (LBCL). The LBCL may be relapsed and / or refractory LBCL. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of diffuse large B-cell lymphoma (DLBCL). The DLBCL may be relapsed and / or refractory DLBCL. The DLBCL may be de novo or transformed. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of high grade B cell lymphoma (HGBCL). The lymphoma may be a double / triple hit high grade B cell lymphoma (HGBCL). The HGBCL may be a relapsed and / or refractory HGBCL. The HGBCL may be a relapsed and / or refractory double / triple hit HGBCL. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of primary mediastinal large B cell lymphoma (PMBCL). The PMBCL may be a relapsed and / or refractory PMBCL. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of follicular lymphoma. The FL may be a relapsed and / or refractory FL. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of follicular lymphoma grade 3B (FL3B). The FL3B may be relapsed and / or refractory FL3B. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of mantle cell lymphoma (MCL). The MCL may be relapsed and / or refractory MCL.In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of small lymphocytic lymphoma (SLL). The SLL may be relapsed and / or refractory SLL. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of marginal zone lymphoma (MZL) (e.g., nodal, extranodal or mucosal-associated). The MZL may be relapsed and / or refractory MZL. Optionally, in each of the above cases, the treatment may be after a previous CAR-T therapy, e.g., a CD19-directed CAR-T therapy. In an alternative option, the treatment may be in the absence of a previous CAR-T therapy, e.g., a CD19-directed CAR-T therapy. Optionally, in each of the above cases, the treatment may be after treatment with a chemotherapy regimen including a CD20 monoclonal antibody. Further, optionally, in each of the above cases, the treatment may be administered after a prior autologous hematopoietic stem cell transplant (HSCT). Optionally, the treatment may be used when the subject's Eastern Cooperative Oncology Group (ECOG) activity status may be 2 or less.

[0020] In some embodiments, the present invention relates to a method of treating a subject having relapsed and / or refractory NHL (e.g., DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof, where the relapsed and / or refractory NHL may have relapsed after or failed to respond to at least two previous treatment regimens, including chemotherapy regimens comprising an anti-CD20 monoclonal antibody. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of relapsed and / or refractory NHL (e.g., DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL), where the relapsed and / or refractory may have relapsed after or failed to respond to at least two previous treatment regimens, including chemotherapy regimens comprising an anti-CD20 monoclonal antibody.

[0021] In some embodiments, the present invention relates to a method of treating a subject with relapsed and / or refractory large B-cell lymphoma (DLBCL, HGBCL, PMBCL, FL3B), where the relapsed and / or refractory may include failure of a previous autologous hematopoietic stem cell transplant (HSCT). In some embodiments, the present invention relates to a method of treating a subject with relapsed and / or refractory large B-cell lymphoma (DLBCL, HGBCL, PMBCL, FL3B), where the subject is ineligible for or unable to undergo an autologous stem cell transplant due to age and / or comorbidities. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of relapsed and / or refractory large B-cell lymphoma (DLBCL, HGBCL, PMBCL, FL3B), where the relapsed and / or refractory may include failure of a previous autologous hematopoietic stem cell transplant (HSCT). In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of relapsed and / or refractory large B-cell lymphoma (DLBCL, HGBCL, PMBCL, FL3B) in subjects who are ineligible or unable to undergo autologous stem cell transplantation due to age and / or comorbidities.

[0022] In some embodiments, the present invention relates to a method of treating a subject having NHL (e.g., DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof, the subject may have at least one bidimensionally measurable nodal lesion or one bidimensionally measurable extranodal lesion as measured by a positron emission tomography-computed tomography (PET / CT) scan. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of NHL (e.g., DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL), the subject may have at least one bidimensionally measurable nodal lesion or one bidimensionally measurable extranodal lesion as measured by a positron emission tomography-computed tomography (PET / CT) scan.

[0023] In some embodiments, the present invention relates to a method of treating a subject with acute lymphoblastic leukemia (ALL) by administering a therapeutically effective amount of an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. The ALL may be relapsed and / or refractory ALL. The ALL may be relapsed and / or refractory B-cell acute lymphoblastic leukemia (R / R B-ALL). The ALL may be relapsed and / or refractory CD19-positive B-ALL. Optionally, the treatment may be after a previous CD19-directed CAR-T therapy. In an alternative option, the treatment may be in the absence of a previous CD19-directed CAR-T therapy. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of acute lymphoblastic leukemia (ALL). The ALL may be relapsed and / or refractory ALL. The ALL may be relapsed and / or refractory B-cell acute lymphoblastic leukemia (R / R B-ALL). The ALL may be relapsed and / or refractory CD19-positive B-ALL. Optionally, the treatment may be after a previous CD19-directed CAR-T therapy. In the alternative, the treatment may be in the absence of a previous CD19-directed CAR-T therapy.

[0024] In some embodiments, the present invention relates to a method of treating a subject with acute lymphoblastic leukemia (ALL) by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof, where morphological disease is manifested in the bone marrow (≧5% blasts). In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of acute lymphoblastic leukemia (ALL), where morphological disease is manifested in the bone marrow (≧5% blasts).

[0025] In some embodiments, the present invention relates to a method of treating a subject with refractory and / or relapsed CD19-positive B-ALL by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of refractory and / or relapsed CD19-positive B-ALL. Refractory and / or relapsed CD19-positive B-ALL may comprise at least one of the following criteria: i) After at least two lines of systemic therapy ii) Relapse or refractory or refractory relapse at any time after first salvage therapy iii) relapse at any time after hematopoietic stem cell transplantation (HSCT) iv) Refractory to, intolerant to, or ineligible / unable to receive SOC treatment options, including blinatumomab and inotuzumab v) Patients with R / R B-ALL Ph+ disease and who are intolerant or refractory to available tyrosine kinase inhibitors (TKIs).

[0026] The anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered Q1W or Q2W. The anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered intravenously or subcutaneously. The anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered with an initial priming dose followed by a main dose. The anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered as a lyophilizate.

[0027] The anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered at 0.1 μg / kg (100 ng / kg) Q1W dosing, optionally intravenously. The anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered at a dose selected from the group consisting of 0.1, 0.3, 1, 3, 10, 20, 40, 80, 160, 320, 640, 1280, 2560 micrograms per kilogram (μg / kg). The dose is based on the subject's weight measurement in kg. The anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered with an initial priming dose, for example, 80 μg / kg, followed by a main dose of 160 μg / kg.

[0028] In some embodiments, the anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered with one or more agents selected from the group consisting of tocilizumab, siltuximab, cyclophosphamide, anti-thymocyte globulin (ATG), alemtuzumab, and anakinra. In some embodiments, the anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered with one or more agents selected from the group consisting of steroids, anti-IL-6 antibodies, anti-TNF antibodies, and anti-IL-1R antibodies. In some embodiments, the anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered with one or more agents selected from the group consisting of tocilizumab, siltuximab, cyclophosphamide, anti-thymocyte globulin (ATG), alemtuzumab, anakinra, steroids, anti-IL-6 antibodies, anti-TNF antibodies, and anti-IL-1R antibodies. In some embodiments, the anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered with one or more agents selected from the group consisting of antihistamines, steroids (including corticosteriods), or other anti-T cell directed therapies (e.g., tocilizumab or canakinumab). In some embodiments, the anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered with tocilizumab and / or corticosteriods. For example, the steroids may be prednisone or dexamethasone. In some embodiments, the anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered with a CRS therapy.

[0029] In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of a subject having a condition selected from the group consisting of LBCL, DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL and ALL, wherein the anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of a subject having a condition selected from the group consisting of R / R LBCL, R / R DLBCL, R / R HGBCL, R / R PMBCL, R / R FL, R / R FL3B, R / R MCL, R / R SLL, R / R MZL and R / R ALL, wherein the anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 39. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of a subject with R / R NHL, which may comprise: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 39.In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of subjects with R / R ALL, the anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 39. The anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered Q1W or Q2W. The anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered intravenously or subcutaneously. The anti-CD19×anti-CD3×anti-CD2 trispecific agent may be administered by an initial priming dose followed by a main dose.

[0030] In some embodiments, the invention relates to a method of treating a subject having a CD19-related disease or disorder, comprising administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent at a dosage selected from the group consisting of 0.1, 0.3, 1, 3, 10, 20, 40, 80, 160, 320, 640, 1280, 2560 micrograms per kilogram (μg / kg).

[0031] In some embodiments, the present invention relates to an anti-CD19xanti-CD3xanti-CD2 trispecific agent for use as a medicament, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent is administered at a dosage selected from the group consisting of 0.1, 0.3, 1, 3, 10, 20, 40, 80, 160, 320, 640, 1280, 2560 micrograms per kilogram (μg / kg).

[0032] In some embodiments, an anti-CD19xanti-CD3xanti-CD2 trispecific agent may comprise a CD19 binding moiety having CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, and CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences of SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.

[0033] In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent comprises (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO:37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO:38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO:39. [Brief description of the drawings]

[0034] [Figure 1] FIG. 1 shows the format of the anti-CD19×anti-CD3×anti-CD2 trispecific agent. The anti-CD2 moiety is shown as the CD58 moiety. [Diagram 2] CD19TSP1 mediates dose-dependent killing of Karpas-422 cells. [Diagram 3] CD19TSP1 induces dose-dependent T cell proliferation in a Karpas-422 and T cell co-culture system. [Figure 4] CD19TSP1 treatment results in secretion of IFNγ, IL-2 and TNFα from Karpas422-T cell co-cultures. [Diagram 5] CD19TSP1 demonstrated dose-dependent antitumor activity in an AdT model with DLBCL xenografts. [Figure 6] CD19TSP1 induced a sustained decrease in peripheral blood B cell numbers in cynomolgus monkeys. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0035] As used herein, the following terms are intended to have the following meanings:

[0036] Antibody: The term "antibody" as used herein refers to a polypeptide (or set of polypeptides) of the immunoglobulin family capable of non-covalently, reversibly and specifically binding to an antigen. For example, a naturally occurring "antibody" of the IgG type is a tetramer comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain is composed of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region is composed of three domains, CH1, CH2 and CH3. Each light chain is composed of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is composed of one domain (abbreviated herein as CL). The VH and VL regions can be further subdivided into hypervariable regions called complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant region of the antibody may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. The term "antibody" includes, but is not limited to, monoclonal antibodies, human antibodies, humanized antibodies, camelized antibodies, chimeric antibodies, bispecific or multispecific antibodies, and anti-idiotypic (anti-Id) antibodies (including, for example, anti-Id antibodies to the antibodies of the present disclosure). The antibody can be of any isotype / class (e.g., IgG, IgE, IgM, IgD, IgA and IgY) or subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2).

[0037] Both light and heavy chains are divided into regions of structural and functional homology. The terms "constant" and "variable" are used functionally. In this regard, it will be understood that the variable domains of both the light (VL) and heavy (VH) chain portions determine antigen recognition and specificity. Conversely, the constant domains of the light (CL) and heavy chains (CH1, CH2 or CH3) confer important biological properties such as secretion, transplacental mobility, Fc receptor binding, complement binding, etc. By convention, the numbering of the constant region domains increases the more distal they are from the antigen binding site or amino terminus of the antibody. In wild-type antibodies, at the N-terminus are the variable regions and at the C-terminus are the constant regions; the CH3 and CL domains actually comprise the carboxy termini of the heavy and light chains, respectively.

[0038] Antibody fragment: The term "antibody fragment" of an antibody as used herein refers to one or more portions of an antibody. In certain embodiments, these portions are part of the contact domain of an antibody. In certain other embodiments, these portions are antigen-binding fragments that retain the ability to bind non-covalently, reversibly and specifically to an antigen, sometimes referred to herein as "antigen-binding fragments", "antigen-binding fragments thereof", "antigen-binding portions", etc. Examples of binding fragments include, but are not limited to, single chain Fv (scFv), Fab fragments, monovalent fragments consisting of the VL, VH, CL and CH1 domains; F(ab)2 fragments, bivalent fragments comprising two Fab fragments linked by a disulfide bridge at the hinge region; Fd fragments consisting of the VH and CH1 domains; Fv fragments consisting of the VL and VH domains of a single arm of an antibody; dAb fragments consisting of the VH domain (Ward et al., 1989, Nature 341:544-546); and isolated complementarity determining regions (CDRs). Thus, the term "antibody fragment" encompasses proteolytic fragments of antibodies (eg, Fab and F(ab)2 fragments) as well as modified proteins comprising one or more portions of an antibody (eg, scFv).

[0039] Antibody fragments can also be incorporated into single domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NARs, and bis-scFvs (see, e.g., Hollinger and Hudson, 2005, Nature Biotechnology 23:1126-1136). Antibody fragments can be grafted into scaffolds based on polypeptides such as fibronectin type III (Fn3) (see U.S. Pat. No. 6,703,199, which describes fibronectin polypeptide monobodies).

[0040] Antibody fragments can be incorporated into single-chain molecules comprising a pair of tandem Fv segments (e.g., VH-CH1-VH-CH1), which combine with complementary light chain polypeptides (e.g., VL-VC-VL-VC) to form a pair of antigen-binding regions (Zapata et al., 1995, Protein Eng. 8:1057-1062; and U.S. Pat. No. 5,641,870).

[0041] Antibody numbering system: In this specification, references to numbered amino acid residues in antibody domains are based on the EU numbering system (e.g., in Table 1) unless otherwise specified. This system was first devised by Edelman et al., 1969, Proc. Nat'l Acad. Sci. USA 63:78-85, and is described in detail in Kabat et al., 1991, in Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, NIH, USA.

[0042] The term "anti-CD19 x anti-CD3 x anti-CD2 trispecific agent" refers to an agent (eg, a therapeutic agent) that targets CD19, CD3 and CD2.

[0043] B cell malignancies: As used herein, B cell malignancies refers to the uncontrolled proliferation of B cells. Examples of B cell malignancies include non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma, leukemia, and myeloma. For example, the B cell malignancy can be, but is not limited to, multiple myeloma, chronic lymphocytic leukemia (CLL) / small lymphocytic lymphoma (SLL), large B cell lymphoma (LBCL), follicular lymphoma (FL), follicular lymphoma grade 3B (FL3B), mantle cell lymphoma (MCL), diffuse large B cell lymphoma (DLBCL), high grade B cell lymphoma (HGBCL), primary mediastinal large B cell lymphoma (PMBCL), marginal zone lymphoma (MZL), Burkitt's lymphoma, lymphoplasmacytic lymphoma (Waldenstrom's macroglobulinemia), hairy cell leukemia, splenic marginal zone B cell lymphoma, extranodal marginal zone lymphoma (EMZL), nodal marginal zone B cell lymphoma (NZML), and primary effusion lymphoma.

[0044] Cancer: The term "cancer" refers to a disease characterized by uncontrolled (often rapid) growth of abnormal cells. Cancer cells can spread locally or via the bloodstream and lymphatic system to other parts of the body. Examples of cancer include the B cell malignancies described herein. The term "cancerous B cell" refers to a B cell that is undergoing or has undergone uncontrolled proliferation.

[0045] CD2: The term "CD2" refers to the cluster of differentiation 2 molecule. It can be found on T cells or natural killer cells. CD2 interacts with lymphocyte function-associated antigen CD58 (LFA-3) and CD48 / BCM1, mediating adhesion between T cells and other cell types. CD2 is involved in T cell development, and the cytoplasmic domain is involved in signaling function. Human and mouse amino acid and nucleic acid sequences can be found in public databases, such as GenBank, UniProt, and Swiss-Prot. For example, the amino acid sequence of human CD2 can be found under UniProt / Swiss-Prot accession number P06729, and the nucleotide sequence encoding human CD2 can be found under accession number NM_001767.5.

[0046] CD3: The term "CD3" or "cluster of differentiation 3" refers to a specialized tri-coreceptor group of T cell receptors. CD3 promotes activation of both cytotoxic T cells (e.g., CD8+ naive T cells) and T helper cells (e.g., CD4+ naive T cells) and is composed of four distinct chains: one CD3γ chain (e.g., Genbank accession numbers NM_000073 and MP_000064 (human)), one CD3δ chain (e.g., Genbank accession numbers NM_000732, NM_001040651, NP_00732 and NP_001035741 (human)) and two CD3ε chains (e.g., Genbank accession numbers NM_000733 and NP_00724 (human)). The chains of CD3 are highly related cell surface proteins of the immunoglobulin superfamily that contain a single extracellular immunoglobulin domain. The CD3 molecule associates with the T cell receptor (TCR) and the ζ-chain to form the T cell receptor (TCR) complex that functions in generating activation signals in T lymphocytes. Unless expressly indicated, reference to CD3 in this application may refer to the CD3 coreceptor, the CD3 coreceptor complex, or any polypeptide chain of the CD3 coreceptor complex.

[0047] CD19: The term "CD19" or "cluster of differentiation 19" refers to the cluster of differentiation 19 protein, an antigenic determinant detectable on leukemic progenitor cells. Human and mouse amino acid and nucleic acid sequences can be found in public databases such as GenBank, UniProt and Swiss-Prot. For example, the amino acid sequence of human CD19 can be found under UniProt / Swiss-Prot accession number P15391, and the nucleotide sequence encoding human CD19 can be found under accession number NM_001178098. CD19 is expressed in most cancers of the B-cell lineage, including, for example, acute lymphoblastic leukemia, chronic lymphocytic leukemia (CLL) / small lymphocytic lymphoma (SLL) and non-Hodgkin's lymphoma. It is also an early marker of B-cell progenitor cells. See, e.g., Nicholson et al., 1997, Mol. Immun. 34(16-17):1157-1165.

[0048] Chimeric antigen receptor: The term "chimeric antigen receptor" or alternatively "CAR" refers to a set of polypeptides, typically two in the simplest embodiment, that when present in an immune effector cell provide the cell with specificity for a target cell, typically a cancer cell, and with intracellular signal generation. In some embodiments, the CAR comprises at least an extracellular antigen binding domain, a transmembrane domain, and a cytoplasmic signaling domain (also referred to herein as an "intracellular signaling domain") that comprises a functional signaling domain derived from a stimulatory molecule and / or a costimulatory molecule as defined below. The set of polypeptides may or may not be contiguous with each other. If the polypeptides are not contiguous with each other, the set of polypeptides comprises a dimerization switch that allows the polypeptides to bind to each other upon the presence of a dimerization molecule, e.g., the antigen binding domain to be linked to the intracellular signaling domain. CAR molecules are typically administered to a subject by administration of immune effector cells (e.g., T cells, preferably autologous to the subject) engineered to express the CAR molecule.

[0049] In combination: Administered in "combination," as used herein, means that two (or more) different therapies are delivered to a subject while the subject is suffering from a disorder, for example, the two or more therapies are delivered after the subject is diagnosed with a disorder and before the disorder is cured or eliminated or the treatment is discontinued for other reasons. The terms "combination" or "in combination with" are not intended to imply that the therapies or therapies must be administered simultaneously and / or formulated to be delivered together, but that these delivery methods are within the scope described herein. The combined therapeutic agent may be administered simultaneously with, before, or after one or more other additional therapies or therapies. The therapeutic agents or treatment protocols may be administered in any order. Generally, each agent will be administered at a dosage and / or time schedule determined for that agent. It will further be understood that the additional therapeutic agents used in the combination may be administered together or separately in different compositions. In general, it is expected that the additional therapeutic agents used in the combination will be used at levels that do not exceed the levels at which they are used individually. In some embodiments, the levels used in combination will be lower than the levels used individually.

[0050] Complementarity determining region: As used herein, the term "complementarity determining region" or "CDR" refers to a sequence of amino acids in an antibody variable region that confers antigen specificity and binding affinity. For example, there are generally three CDRs in each heavy chain variable region (e.g., CDR-H1, CDR-H2, and CDR-H3) and three CDRs in each light chain variable region (CDR-L1, CDR-L2, and CDR-L3). The precise amino acid sequence boundaries of a given CDR can be determined using the "Kabat" numbering scheme, Kabat et al., 1991, "Sequences of Proteins of Immunological Interest", 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme), Al-Lazikani et al., 1997, JMB 273, 927-948 ("Chothia" numbering scheme) and the ImMunoGenTics (IMGT) numbering scheme (Lefranc, 1999, the Immunologist 7:132-136; Lefranc, et al. The CDR numbering scheme may be determined using any of several well-known schemes, including those described by Kabat et al., 2003, Dev. Comp. Immunol. 27:55-77 (the "IMGT" numbering scheme). For example, for the classical format, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (CDR-H1), 50-65 (CDR-H2), and 95-102 (CDR-H3); According to Chothia, the CDR amino acid residues in VH are numbered 26-32 (CDR-H1), 52-56 (CDR-H2) and 95-102 (CDR-H3); the amino acid residues in VL are numbered 26-32 (CDR-L1), 50-52 (CDR-L2) and 91-96 (CDR-L3).Combining both Kabat and Chothia CDR definitions, the CDRs consist of amino acid residues 26-35 (CDR-H1), 50-65 (CDR-H2) and 95-102 (CDR-H3) in human VH and amino acid residues 24-34 (CDR-L1), 50-56 (CDR-L2) and 89-97 (CDR-L3) in human VL. According to IMGT, the CDR amino acid residues in VH are numbered approximately 26-35 (CDR-H1), 51-57 (CDR-H2) and 93-102 (CDR-H3), and the CDR amino acid residues in VL are numbered approximately 27-32 (CDR-L1), 50-52 (CDR-L2) and 89-97 (CDR-L3) ("Kabat" numbering). With IMGT, the CDR regions of an antibody can be determined using the program IMGT / Domain Gap Align.

[0051] ECOG activity status score: The term "ECOG activity status score" as used herein refers to a subject's score on the Eastern Cooperative Oncology Group (ECOG) activity status score as described in Oken et al., 1982, Am J Clin Oncol 5(6):649-55. The score can range from 0 to 5.

[0052] [Table 1]

[0053] Effective amount: The term "effective amount" or "therapeutically effective amount" or "pharmaceutical effective amount" refers to an amount or quantity of an active agent (or combination of agents) that is sufficient to elicit the required or desired response when administered to a subject, or in other words, an amount that is sufficient to elicit a significant biological response. Said amount preferably relates to an amount that is therapeutically, or more broadly, prophylactically effective against the progression of a disease or disorder disclosed herein. It is understood that an "effective amount" or a "therapeutically effective amount" may vary between subjects due to variability in the metabolism of the drug, the age, weight, general condition of the subject, the condition being treated, the severity of the condition being treated, and the judgment of the prescribing physician.

[0054] Half antibody: The term "half antibody" refers to a molecule that comprises at least one antigen binding module (ABM) or ABM chain, where the ABM has the ability to non-covalently, reversibly and specifically bind to an antigen. A half antibody may associate with another molecule that comprises an ABM or ABM chain, for example, through disulfide bridges or molecular interactions (e.g., knob-in-hole interactions between Fc heterodimers). A half antibody may be composed of one polypeptide chain or two or more polypeptide chains (e.g., two polypeptide chains of a Fab). In one embodiment, a half antibody comprises an Fc region.

[0055] An example of a half antibody is a molecule comprising the heavy and light chains of an antibody (e.g., an IgG antibody). Another example of a half antibody is a molecule comprising a first polypeptide comprising a VL domain and a CL domain, and a second polypeptide comprising a VH domain, a CH1 domain, a hinge region, a CH2 domain, and a CH3 domain, where the VL and VH domains form an ABM. Yet another example of a half antibody is a polypeptide comprising an scFv domain, a CH2 domain, and a CH3 domain.

[0056] A half antibody can comprise two or more ABMs, for example, a half antibody comprises (in order from N-terminus to C-terminus) an scFv domain, a CH2 domain, a CH3 domain and another scFv domain.

[0057] A half antibody may also comprise an ABM chain that, when associated with another ABM chain in another half antibody, forms a complete ABM.

[0058] Single-chain Fab or scFab: The terms "single-chain Fab" and "scFab" refer to a polypeptide comprising an antibody heavy chain variable domain (VH), an antibody constant domain 1 (CH1), an antibody light chain variable domain (VL), an antibody light chain constant domain (CL) and a linker, such that VH and VL are associated with each other and CH1 and CL are associated with each other. In certain embodiments, the antibody domains and linkers have one of the following orders from N-terminus to C-terminus: a) VH-CH1-linker-VL-CL, b) VL-CL-linker-VH-CH1, c) VH-CL-linker-VL-CH1 or d) VL-CH1-linker-VH-CL. The linker can be a polypeptide of at least 30 amino acids, for example, 32-50 amino acids. Single-chain Fabs are stabilized by a native disulfide bond between the CL and CH1 domains.

[0059] Single-chain Fv or scFv: The term "single-chain Fv" or "scFv" as used herein refers to an antibody fragment comprising the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. The Fv polypeptide may further comprise a polypeptide linker between the VH and VL domains that enables the scFv to form the desired structure for antigen binding. For a review of scFvs, see Plueckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., 1994, Springer-Verlag, New York, pp. 269-315.

[0060] Specific (or selective) binding: The term "specifically (or selectively) binding" to an antigen or epitope refers to a binding reaction that determines the presence of the same antigen or epitope in a heterogeneous population of proteins and other biological substances. ABMs are typically 5×10 -2 Less than M, 10 -2 Less than M, 5×10 -3 Less than M, 10 -3 Less than M, 5×10 -4 Less than M, 10-4 Less than M, 5×10 -5 Less than M, 10 -5 Less than M, 5×10 -6 Less than M, 10 -6 Less than M, 5×10 -7 Less than M, 10 -7 Less than M, 5×10 -8 Less than M, 10 -8 Less than M, 5×10 -9 Less than M or 10 -9 It also has a dissociation rate constant (KD) (koff / kon) less than M and binds to the target antigen with an affinity at least 2-fold higher than its affinity for binding to a non-specific antigen (e.g., HSA). Binding affinity can be measured using Biacore, SPR, or BLI assays. The term "specifically binds" does not exclude cross-species cross-reactivity. For example, an antigen-binding module (e.g., an antigen-binding fragment of an antibody) that "specifically binds" to an antigen from one species may also "specifically bind" to the corresponding antigen in one or more other species. Thus, such cross-species cross-reactivity does not itself change the classification of the antigen-binding module as a "specific" binder. In certain embodiments, an antigen binding module that specifically binds to a human antigen has cross-species reactivity with one or more non-human mammalian species, such as a primate species (including, but not limited to, one or more of Macaca fascicularis, Macaca mulatta, and Macaca nemestrina) or a rodent species, such as Mus musculus. In other embodiments, the antigen binding module does not have cross-species reactivity.

[0061] Subject: The term "subject" includes human and non-human animals. Non-human animals include all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, and reptiles. Unless otherwise specified, the terms "patient" and "subject" are used interchangeably herein.

[0062] Therapeutically effective amount: A "therapeutically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result.

[0063] Treat, Treatment, Treating: As used herein, the terms "treat", "treatment" and "treating" refer to a reduction or amelioration of the progression, severity and / or duration of a disease or disorder (e.g., B-cell malignancies) or the amelioration of the progression, severity and / or duration of one or more symptoms (e.g., one or more identifiable symptoms) of a disorder (e.g., CRS) resulting from administration of one or more anti-CD19 agents. In certain embodiments, the terms "treat", "treatment" and "treating" refer to an improvement in at least one measurable physical parameter of a disease, such as tumor growth, not necessarily discernible by the patient. In other embodiments, the terms "treat", "treatment" and "treating" refer to an inhibition of progression of a disease, either physically, e.g., by stabilization of an identifiable symptom, physiologically, e.g., by stabilization of a physical parameter, or both. In some embodiments, the terms "treat", "treatment" and "treating" may refer to a reduction or stabilization of tumor size or cancerous cell number.

[0064] Trispecific binding molecule: The term "trispecific binding molecule" or "TBM" refers to a molecule that specifically binds to three antigens and comprises three or more antigen-binding domains. The TBM of the present disclosure comprises at least one antigen-binding domain specific for CD19, at least one antigen-binding domain specific for CD3, and at least one antigen-binding domain specific for CD2.

[0065] VH: The term "VH" refers to the variable region of an immunoglobulin heavy chain of an antibody, including the heavy chain of an Fv, scFv, dsFv or Fab.

[0066] VL: The term "VL" refers to the variable region of an immunoglobulin light chain, including the light chain of an Fv, scFv, dsFv or Fab.

[0067] Anti-CD19 × anti-CD3 × anti-CD2 trispecific agents Disclosed herein are anti-CD19×anti-CD3×anti-CD2 trispecific agents that, at least in part, target CD19+ cells (malignant B cells as well as normal B cells and follicular dendritic cells) and engage CD3 (TCR signaling component) and CD2 (costimulatory receptor) on T cells, thereby triggering cytotoxicity of redirected T cells against CD19-positive malignant cells.

[0068] It is believed that the simultaneous binding of anti-CD19×anti-CD3×anti-CD2 trispecific agents to multiple molecules, including CD19 on malignant B cells, the CD3 subunit of the T cell receptor (TCR) complex, and CD2 on T cells, leads to TCR cross-linking and CD2 costimulation, followed by the formation of a cytolytic immune synapse, resulting in T cell activation and specific lysis of malignant B cells. Without being bound by theory, it is believed that the association of CD2 with CD3 provides a costimulatory signal that efficiently activates T cells and leads to sustained T cell responses. Such costimulatory signals may improve T cell responses to tumors and help overcome the limitations of CD3 bispecific antibodies.

[0069] The sequences associated with anti-CD19 x anti-CD3 x anti-CD2 trispecific agents are shown in the table below. Anti-CD19 x anti-CD3 x anti-CD2 trispecific agents and methods for making the same are also disclosed in International Application PCT / US2020 / 033559 (WO 2020 / 236792), which is incorporated herein by reference. The anti-CD19 x anti-CD3 x anti-CD2 trispecific agent may be CD3hi TSP1 (H variant) in Table 19C in International Application PCT / US2020 / 033559 (WO 2020 / 236792), which is incorporated herein by reference. The first half antibody heavy chain comprises a CD19 binding sequence and a CD3 binding sequence. The first half antibody light chain comprises a CD19 binding sequence. The second half antibody comprises a CD2 binding sequence (the CD2 binding sequence may be a portion of a CD58 sequence). See, for example, Figure 1, anti-CD19 x anti-CD3 x anti-CD2 trispecific agent format, where the anti-CD2 moiety is shown as the CD58 moiety.

[0070]

Table 2

[0071]

Table 3

[0072]

Table 4

[0073]

Table 5

[0074]

Table 6

[0075]

Table 7

[0076]

Table 8

[0077]

Table 9

[0078]

Table 10

[0079] In some embodiments, an anti-CD19xanti-CD3xanti-CD2 trispecific agent may comprise a CD19 binding moiety having CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, and CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences of SEQ ID NO:14, SEQ ID NO:15, and SEQ ID NO:16.

[0080] In some embodiments, an anti-CD19xanti-CD3xanti-CD2 trispecific agent may comprise a CD19 binding moiety having CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, and CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences of SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.

[0081] In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD19 binding moiety having a VH having the amino acid sequence of SEQ ID NO: 13. The CD19 binding moiety may also comprise a VL having the amino acid sequence of SEQ ID NO: 26. The CD19 binding moiety may also comprise both a VH having the amino acid sequence of SEQ ID NO: 13 and a VL having the amino acid sequence of SEQ ID NO: 26. In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD19 binding moiety comprising any of the amino acid sequences in Table 1.

[0082] In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD3 binding moiety having CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NO:27, SEQ ID NO:28, and SEQ ID NO:29, and CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences of SEQ ID NO:30, SEQ ID NO:31, and SEQ ID NO:32.

[0083] In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD3 binding moiety comprising a VH having the amino acid sequence of SEQ ID NO: 33. In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD3 binding moiety comprising a VL having the amino acid sequence of SEQ ID NO: 34. In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD3 binding moiety comprising the amino acid sequence of SEQ ID NO: 35. In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD3 binding moiety comprising any of the amino acid sequences given in Table 2 and / or Table 3.

[0084] In some embodiments, the CD2 binding moiety domain comprises the CD2 interacting IgV domain of its natural ligand CD58 (CD58-IgV, also referred to as anti-CD2). In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD2 binding moiety that is a CD58 moiety. The CD58 moiety may comprise amino acid residues 30-123 of full length wild type CD58. In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD2 binding moiety having the amino acid sequence of SEQ ID NO: 36. In some embodiments, an anti-CD19×anti-CD3×anti-CD2 trispecific agent may comprise a CD2 binding moiety comprising any of the amino acid sequences given in Table 4.

[0085] B-cell malignancies The CD19 protein is broadly expressed in the B lymphocyte lineage. CD19 expression is maintained in B lineage cells undergoing neoplastic transformation to ALL and B-NHL (Scheumermann and Racila 1995).

[0086] In some embodiments, the B-cell malignancy is B-cell acute lymphocytic leukemia (also known as B-cell acute lymphoblastic leukemia or B-cell acute lymphocytic leukemia) (ALL or B-ALL), e.g., relapsed and / or refractory B-ALL.

[0087] In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of relapsed and / or refractory B-NHL. In some embodiments, the present invention relates to a method of treating a subject with relapsed and / or refractory LBCL by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof, wherein the subject has previously received CAR-T therapy. In some embodiments, the present invention relates to a method of treating a subject with relapsed and / or refractory LBCL by administering an anti-CD19×anti-CD3×anti-CD2 trispecific agent to a subject in need thereof, wherein the subject has not previously received CAR-T therapy. In some embodiments, the present invention relates to an anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of relapsed and / or refractory B-ALL.

[0088] In some embodiments, the B cell malignancy is chronic lymphocytic leukemia (CLL) / small lymphocytic lymphoma (SLL), e.g., relapsed and / or refractory CLL / SLL.

[0089] In some embodiments, the B cell malignancy is marginal zone lymphoma (MZL).

[0090] In some embodiments, the B cell malignancy is extranodal marginal zone lymphoma (EMZL).

[0091] In some embodiments, the B cell malignancy is nodal marginal zone B cell lymphoma (NZML).

[0092] In some embodiments, the B cell malignancy is splenic marginal zone B cell lymphoma (SMZL).

[0093] Refractory or relapsed CD19-positive B-ALL may include at least one of the following: i) At least two lines of systemic therapy ii) Relapse or refractory or refractory relapse at any time after first salvage therapy iii) relapse at any time after hematopoietic stem cell transplantation (HSCT) iv) Refractory to, intolerant to, or ineligible / unable to receive SOC treatment options, including blinatumomab and inotuzumab v) Patients with R / R B-ALL Ph+ disease and who are intolerant or refractory to available tyrosine kinase inhibitors (TKIs).

[0094] The endpoint and / or treatment may be measured in terms of one or more of the following criteria: 1) Incidence and severity of dose-limiting toxicities (DLTs) (e.g., after 28 or 35 days, depending on dosing schedule) 2) Incidence and severity of adverse events (AEs) and serious adverse events (SAEs) (e.g., after 21 months) 3) Frequency of treatment interruption (e.g., after 21 months) 4) Frequency of dose reduction (e.g., after 21 months) 5) Dose intensity (e.g., after 21 months)

[0095] Furthermore, the endpoint and / or treatment may be measured in terms of one or more of the following criteria: 1) Objective response rate (ORR) (e.g., after 21 months) 2) Complete remission (CR) rate (e.g., after 21 months) 3) Best Overall Response (BOR) (e.g., after 21 months) 4) Duration of response (DOR) (e.g., after 21 months) 5) Overall survival (OS) (e.g., after 33 months) 6) Progression-free survival (PFS) (e.g., after 21 months) 7) Event-free survival (EFS) (e.g., after 21 months) 8) Maximum concentration (Cmax) of CD19TSP1 (e.g., after 21 months) 9) Area under the curve (AUC) of CD19TSP1 (e.g., after 21 months) 10) CD19TSP1 trough concentration (C trough) (e.g., after 21 months) 11) Prevalence of anti-drug antibodies (ADA) (e.g., at baseline) 12) Incidence of anti-drug antibodies (ADA) during treatment (e.g., after 21 months)

[0096] Treatment may improve one or more of the following criteria in combination with an acceptable safety profile. i) Remission rate ii) Duration of remission iii) overall survival

[0097] In some embodiments, one or more of the agents selected from the group consisting of tocilizumab, siltuximab, cyclophosphamide, anti-thymocyte globulin (ATG), alemtuzumab, anakinra, steroids, anti-IL-6, anti-TNF and anti-IL-1R antibodies, antihistamines, steroids (including corticosteriods), canakinumab may be administered after the anti-CD19×anti-CD3×anti-CD2 trispecific agent. For example, the steroid may be prednisone or dexamethasone. In some embodiments, administration of the anti-CD19×anti-CD3×anti-CD2 trispecific agent may be after or concurrent with acetaminophen and / or diphenhydramine. In some embodiments, administration of the anti-CD19×anti-CD3×anti-CD2 trispecific agent may be before, after or concurrent with CRS therapy.

[0098] Pharmaceutical Compositions and Combination Administration The anti-CD19×anti-CD3×anti-CD2 trispecific agent can be formulated as a pharmaceutical composition containing one or more pharma- ceutically acceptable excipients or carriers. To prepare a pharmaceutical or sterile composition, the formulation of the anti-CD19×anti-CD3×anti-CD2 trispecific agent can be combined with one or more pharma- ceutically acceptable excipients and / or carriers. The composition can be in various forms. These include, for example, liquid, semi-solid and solid dosage forms, such as solutions (e.g., injectable and infusible solutions), dispersions or suspensions, liposomes and suppositories. The preferred form depends on the intended mode of administration and therapeutic application. Exemplary preferred compositions are in the form of injectable or infusible solutions. The preferred mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In a preferred embodiment, the anti-CD19×anti-CD3×anti-CD2 trispecific agent is administered by intravenous infusion or injection. In another preferred embodiment, the anti-CD19 x anti-CD3 x anti-CD2 trispecific agent is administered by subcutaneous injection. Therapeutic compositions typically need to be sterile and stable under the conditions of manufacture and storage. The compositions can be formulated as solutions, microemulsions, dispersions, liposomes, or other ordered structures suitable for high antibody concentration. Sterile injectable solutions can be prepared by incorporating the active compound (i.e., antibody or antibody moiety) in the required amount in the appropriate solvent, followed by filtered sterilization. Pharmaceutical compositions for use in the disclosed methods may be made in conventional manner. The use of antibodies as active ingredients in pharmaceuticals is now widespread, including products such as Herceptin® (trastuzumab), Rituxan® (rituximab), Synagis® (palivizumab). Techniques for lyophilization, preparation of aqueous formulations, and purification of antibodies to pharmaceutical grade are well known in the art. The antibodies are typically formulated in aqueous form for parenteral administration or as a lyophilizate for reconstitution with a suitable diluent prior to administration. In some embodiments of the disclosed methods and uses, the antibodies of the invention are formulated as a lyophilizate.A suitable lyophilized formulation can be reconstituted in a small liquid volume (e.g., 2 ml or less) to allow for subcutaneous administration, and can result in a low level of antibody aggregation in the solution. For immediate administration, it is dissolved in a suitable aqueous carrier, such as sterile water for injection or sterile buffered saline. If it would be desirable to create a larger volume of solution for administration by infusion, rather than bolus injection, it may be advantageous to incorporate human serum albumin or the patient's own heparinized blood into the saline solution at the time of formulation. The presence of excess of these physiologically inactive proteins prevents antibody reduction by adsorption onto the walls of containers and tubing used with the infusion solution. If albumin is used, the preferred concentration is 0.5-4.5% by weight of the saline solution.

[0099] Method and rate of administration The trispecific agents disclosed herein can be administered by a variety of methods known in the art, although for many therapeutic applications, the preferred route / mode of administration is intravenous injection or infusion. For example, Sachs et al.,Optimal Dosing for Targeted Therapies in Oncology:Drug Development Cases Leading by Example,Clin.Cancer Res;22(6)2016;Bai et al,A Guide to Rational Dosing of Monoclonal Antibodies,Clin.Pharmacokinet.2012:51(2)119-135;Le Tourneau,J.,Dose Escalation Methods in Phase I Cancer Clinical Trials,J Natl Cancer Inst 2009;101:708-720;Wang,D.et al.,Fixed Dosing Versus Body Size-Based Dosing of Monoclonal Antibodies in Adult Clinical Trials,J Clin Pharmacol 2009;29:1012-1024;Hempel,G.et ano,Flat-Fixed Dosing Versus Body Surface Aread-Based Dosing of Anticancer Drugs: There Is a Difference,The Oncologist 2007:12:924-926,Mathijssen,R.See, Leveque, Flat-Fixed Dosing Versus Body Surface Area-Based Dosing of Anticancer Drugs in Adults: Does It Make a Difference?, The Oncologist, 2007;12:913-923; Leveque, Evaluation of Fixed Dosing of New Anticancer Agents in Phase I Studies, Anticancer Research 28:300275-2078(2008), Gurney, How to calculate the dose of chemotherapy, British Journal of Cancer(2002)86,1297-1302; for example, antibody molecules can be administered by intravenous infusion at a rate of greater than 20 mg / min, e.g., 20-40 mg / min, and typically 40 mg / min or more, to achieve a dosage of about 35-440 mg / m2, typically about 70-310 mg / m2, more typically about 110-130 mg / m2. In embodiments, the antibody molecule can be administered by intravenous infusion at a rate of less than 10 mg / min; preferably 5 mg / min or less to achieve a dosage of about 1-100 mg / m2, preferably about 5-50 mg / m2, about 7-25 mg / m2, more preferably about 10 mg / m2. The route and / or mode of administration will vary depending on the desired results.

[0100] Dosage regimen The dosage regimen is adjusted to provide the optimum desired response (e.g., therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time, or the dosage may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. For ease of administration and uniformity of dosage, it is particularly advantageous to formulate parenteral compositions in dosage unit form. Dose unit form, as used herein, refers to a physically discrete unit suitable as a unitary dose to the subject to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the dosage unit form of the present invention are dictated by and directly depend on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the inherent limitations in the art of compounding such active compounds for the treatment of susceptibility in individuals.

[0101] 5. Additional Embodiments All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0102] Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0103] 6. Additional Embodiments Example 1 An in vitro RTCC assay was developed to demonstrate CD19TSP1-mediated specific lysis of the CD19+ B-cell lymphoma cell line Karpas422. Karpas422 cells were modified to overexpress the firefly luciferase gene. Tumor cell death, T cell proliferation and cytokine secretion were assessed after co-culture of Karpas422 cells with T cells in the presence of CD19TSP1 or isotype control antibody TSP1C (having an anti-chicken lysozyme moiety instead of the CD19 binding moiety, i.e., an anti-chicken lysoszyme x CD2 x CD3 targeting antibody). Luciferase signal reduction was then measured as an indicator of Karpas422 cell lysis. Furthermore, T cell activation was assessed by quantifying the production levels of interferon gamma (IFNγ), interleukin-2 (IL-2) and tumor necrosis factor alpha (TNFα) cytokines in the culture supernatants, while T cell proliferation was determined using T cell numbers from flow cytometry.

[0104] T cells from healthy donors were co-cultured with luciferase-expressing Karpas422 cells at a 1:1 ratio for 96 hours in the presence of CD19TSP1 or TSP1C. % killing (% lysis of Karpas422 cells) was determined by the reduction in luciferase activity compared to Karpas422 cells alone. Mean values ​​+ / - SEM were plotted from 12 biological replicates (from 4 individual healthy donor T cells, each repeated in 3 independent experiments). See Figure 2.

[0105] CD19TSP1 at doses above 0.01 nM mediated efficient killing of Karpas422 cells by T cells from healthy human donors in a dose-dependent manner, whereas an isotype control antibody (TSP1C) showed negligible killing at the highest concentration tested, 1 nM.

[0106] T cell proliferation was quantified using flow cytometry.

[0107] CTV-stained T cells from healthy donors were co-cultured with luciferase-expressing Karpas422 cells at a 1:1 ratio for 96 hours in the presence of CD19TSP1 or TSP1C at the indicated concentrations. T cell numbers were determined using flow cytometry by gating on CTV-stained T cells. Mean values ​​+ / - SEM were plotted from 12 biological replicates (from 4 individual healthy donor T cells, each repeated in 3 independent experiments). See Figure 3.

[0108] CD19TSP1 induced concentration-dependent T cell proliferation in the presence of Karpas422 cells. As expected, T cells in the TSP1C control showed no proliferation.

[0109] CD19TSP1 treatment results in the secretion of IFNγ, IL-2 and TNFα from Karpas422-T cell co-culture. Cell culture supernatants were collected from RTCC assays 48 hours after the initiation of Karpas422-T cell co-culture. Secreted IFNγ, IL-2 and TNFα levels were quantified using Custom 3-Plex MSD VPLEX immunoassays. Mean values ​​+ / - SEM were plotted from 12 biological replicates (from 4 individual healthy donor T cells, each repeated in 3 independent experiments). See Figure 4.

[0110] Levels of IFNγ, IL-2 and TNFα in conditioned media from RTCC assays were quantified using a Meso Scale Discovery (MSD) VPLEX multiplex immunoassay. CD19TSP1 treatment of Karpas422 and T cell co-cultures resulted in increased secretion of IFNγ, IL-2 and TNFα, while TSP1C induced zero to low levels of secretion (data not shown). The magnitude of secreted IFNγ was significantly higher than IL-2 and TNFα.

[0111] Example 2 Two preclinical in vivo models were used to evaluate the activity of CD19TSP1: the AdT model and the hCD34+ model. The AdT model was used to evaluate the single-dose activity of CD19TSP1 over a range of doses against established DLBCL tumors. The hCD34+ model was used in a multiple-dose study of CD19TSP1 against established DLBCL tumors. In both model systems, CD19TSP1 demonstrated robust and durable antitumor responses against the same established tumors.

[0112] CD19TSP1 showed dose-dependent antitumor activity and was well tolerated in the AdT model with DLBCL xenografts: on day 0, 5x106 OCILY19 cells were implanted subcutaneously (sc). Two days later, 15 million PBMCs from frozen donor stocks were implanted by IV administration. 11 days after tumor implantation, tumor volumes reached approximately 200 mm3, and animals were randomized to their respective treatment groups and received a single dose of their respective treatment. The antitumor activity of CD19TSP1 was evaluated at different dose levels. Tumor alone groups (in mice without human PBMC adoptive transfer) and tumors (no Ab) in AdT mice were included as controls. CD19TSP1 administered at 0.3 mg / kg, 0.1 mg / kg, and 0.03 mg / kg showed antitumor activity that was statistically different from the control group (p<0.05, one-way ANOVA with post-hoc Dunnett's multiple comparisons). The numbers of CR, PR and NR are as follows for each group: 0.3mg / kg (7CR / 0PR / 1NR), 0.1mg / kg (2CR / 6PR / 0NR) and 0.03mg / kg (2CR / 5PR / 1NR), 0.01mg / kg (1CR / 3PR / 4NR) and 0.003mg / kg (0CR / 7PR / 1NR). CR=complete regression; PR=partial regression; NR=non-response.

[0113] CD19TSP1 showed dose-dependent antitumor activity and durable responses against DLBCL xenografts in hCD34+ models: 5×10 6OCILY19 cells were implanted subcutaneously (sc). Eleven days after implantation, animals were randomized based on tumor size and donor into treatment groups as follows: no treatment, 0.3 mg / kg CD19TSP1, or 0.1 mg / kg CD19TSP1. Animals were treated by IV administration once a week for three weeks (QWx3, IV). CD19TSP1 showed dose-dependent activity. Compared to controls, both 0.3 mg / kg and 0.1 mg / kg CD19TSP1 showed statistically significant antitumor activity (p<0.05, one-way ANOVA with post-hoc SIDAK multiple comparisons). CD19TSP1 administered at 0.3 mg / kg showed robust activity with 5 / 8 CR and 1 PR, while 0.1 mg / kg caused growth retardation and was less active with 1 CR and 1 PR. Both 0.1 mg / kg and 0.3 mg / kg conferred a survival benefit over no treatment. CR = complete regression; PR = partial regression.

[0114] Example 3 Evaluation of CD19TSP1 in a dose-ranging toxicity study in cynomolgus monkeys showed that single and repeated (two doses administered one week apart) intravenous administration of CD19TSP1 was well tolerated. Total B cell numbers were measured in peripheral blood in monkeys treated with a single dose (phase 1) or two doses at one week intervals (phase II) of CD19TSP1 or the trispecific isotype control TSP1C (having an anti-chicken lysozyme moiety instead of the CD19 binding moiety, i.e., an anti-chicken lysoszyme x CD2 x CD3 targeting antibody). Intravenous administration of CD19TSP1 induced sustained B cell depletion in peripheral blood (see Figure 6) and lymphoid organs (based on histopathological evaluation).

[0115] Example 4 CD19TSP1 was well tolerated following a single dose (0.3 mg / kg) subcutaneous administration to cynomolgus monkeys.

[0116] Example 5 Disclosed herein is a Phase I study to characterize the safety and tolerability of anti-CD19×anti-CD3×anti-CD2 trispecific agent (CD19TSP1) and to identify the maximum tolerated dose and / or recommended dose, schedule and route of administration in R / RB cell NHL and / or R / R B-ALL. The study includes a dose escalation part of anti-CD19×anti-CD3×anti-CD2 trispecific agent (CD19TSP1) in R / RB cell NHL and / or R / R B-ALL, and a dose expansion part in i) R / R LBCL and iii) R / R B-ALL that have or have not received CAR-T therapy. During dose escalation, the safety (including dose-dose limiting toxicity (DLT) relationship) and tolerability of anti-CD19×anti-CD3×anti-CD2 trispecific agent (CD19TSP1) will be evaluated, and the schedule, route of administration and dose will be identified for use in the expansion part based on the review of these data. Additionally, the recommended dose will be guided by available information on pharmacokinetics (PK), pharmacodynamics (PD), and preliminary antitumor activity. Dose escalation will be guided by an adaptive Bayesian logistic regression model (BLRM) following the principles of dose escalation with overdose control (EWOC). Different schedules (once weekly (Q1W) or once every two weeks (Q2W) with and without a priming dose) and routes of administration (intravenous (iv) or subcutaneous (sc)) will be explored in the dose escalation group.

[0117] Anti-CD19 x anti-CD3 x anti-CD2 trispecific agent (CD19TSP1) is used in adult NHL patients who have failed two or more lines of chemotherapy and have progressed (or relapsed) after autologous hematopoietic stem cell transplantation (HSCT), or who are ineligible for or do not consent to the procedure.

[0118] The anti-CD19 x anti-CD3 x anti-CD2 trispecific agent (CD19TSP1) is used in adult R / R ALL patients.

[0119] Anti-CD19 x anti-CD3 x anti-CD2 trispecific agent (CD19TSP1) is used in adult R / R LBCL (DLBCL, double / triple hit HGBCL, PMBCL, FL3B patients who have not received CD19-directed CAR-T therapy).

[0120] Anti-CD19 x anti-CD3 x anti-CD2 trispecific agent (CD19TSP1) for use in adult R / R LBCL (DLBCL, double / triple hit HGBCL, PMBCL, FL3B) patients receiving CD19-directed CAR-T therapy.

[0121] The anti-CD19 x anti-CD3 x anti-CD2 trispecific agent (CD19TSP1) is used in adult R / R ALL patients.

[0122] The incidence and severity of dose-limiting toxicity will be used as an assessment of the safety of the study drug. Dose-limiting toxicity will be defined as any CTCAE grade ≥ 3 adverse event or abnormal laboratory value occurring within the DLT evaluation period (28 days for the initial treatment schedule or 35 days for the priming dosing schedule) and not primarily related to disease, disease progression, intercurrent illness, or concomitant medications.

[0123] Overall response rate (ORR), complete response (CR) rate, best overall response (BOR), duration of response (DOR), overall survival (OS), progression-free survival (PFS), and event-free survival (EFS) were based on the Lugano response Criteria Classification criteria for NHL and the National Comprehensive Cancer Network (NCCN) guidelines as relevant for ALL.

[0124] Incorporation by Reference All publications, patents, and accession numbers mentioned in this specification are herein incorporated by reference in their entirety, to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference.

[0125] Equivalent While specific embodiments of the invention have been discussed, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those of skill in the art upon review of this specification and the claims below. The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.

Claims

1. 1. A method of treating a subject having a condition which is non-Hodgkin's lymphoma (NHL) or acute lymphoblastic leukemia (ALL) by administering a therapeutically effective amount of an anti-CD19 x anti-CD3 x anti-CD2 trispecific agent to a subject in need thereof, optionally wherein said NHL or ALL may be relapsed and / or refractory NHL or relapsed and / or refractory ALL.

2. 1. An anti-CD19×anti-CD3×anti-CD2 trispecific agent for use in the treatment of a condition which is non-Hodgkin's lymphoma (NHL) or acute lymphoblastic leukemia (ALL), optionally wherein said NHL or ALL may be relapsed and / or refractory NHL or relapsed and / or refractory ALL.

3. 3. The method of claim 1 or the use of claim 2, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent comprises: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO:

39.

4. 4. The method of claim 1 or 3 or the trispecific agent for use according to claim 2 or 3, wherein the condition is selected from the group consisting of LBCL, DLBCL, HGBCL, PMBCL, FL, FL3B, MCL, SLL, MZL and ALL, and optionally the condition may be relapsed and / or refractory.

5. The method of any one of claims 1 or 3-4 or the trispecific agent for use of any one of claims 2-4, wherein the condition is selected from the group consisting of DLBCL, HGBCL, PMBCL, FL3B, MCL, SLL and MZL, and optionally the condition may be relapsed and / or refractory.

6. The method of any one of claims 1 or 3 to 5 or the trispecific agent for use of any one of claims 2 to 5, wherein the pathology is R / R DLBCL, optionally which is de novo or transformed R / R DLBCL.

7. The method according to any one of claims 1 or 3 to 6 or the trispecific agent for use according to any one of claims 2 to 6, wherein the condition is R / R HGBCL, optionally which is relapsed and / or refractory double / triple hit high grade B-cell lymphoma (HGBCL).

8. The method according to any one of claims 1 or 3 to 7 or the trispecific agent for use according to any one of claims 2 to 7, wherein the treatment may be after a previous CAR-T therapy, and / or after treatment with a chemotherapy regimen comprising a CD20 monoclonal antibody and / or after a previous autologous hematopoietic stem cell transplant (HSCT).

9. The trispecific agent for the method according to any one of claims 1 or 3 to 8 or for use according to any one of claims 2 to 8, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered Q1W or Q2W.

10. The method according to any one of claims 1 or 3 to 9 or the trispecific agent for use according to any one of claims 2 to 9, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered intravenously or subcutaneously.

11. The method according to any one of claims 1 or 3 to 10 or the trispecific agent for use according to any one of claims 2 to 10, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered by an initial priming dose followed by a main dose.

12. The method of any one of claims 1 or 3 to 11 or the trispecific agent for use according to any one of claims 2 to 11, wherein said anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered at a dose selected from the group consisting of: 0.1, 0.3, 1, 3, 10, 20, 40, 80, 160, 320, 640, 1280, 2560 micrograms per kilogram (μg / kg).

13. The method according to any one of claims 1 or 3 to 12 or the trispecific agent for use according to any one of claims 2 to 12, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered before, after or simultaneously with CRS therapy.

14. The method according to any one of claims 1 or 3 to 13 or the trispecific agent for use according to any one of claims 2 to 13, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered before, after or simultaneously with tocilizumab.

15. 1. A method of treating a subject having a CD19-related disease or disorder, comprising administering an anti-CD19 x anti-CD3 x anti-CD2 trispecific agent at a dose selected from the group consisting of 0.1, 0.3, 1, 3, 10, 20, 40, 80, 160, 320, 640, 1280, 2560 micrograms per kilogram (μg / kg).

16. 1. An anti-CD19xanti-CD3xanti-CD2 trispecific agent for use as a medicament, the anti-CD19xanti-CD3xanti-CD2 trispecific agent being administered at a dose selected from the group consisting of: 0.1, 0.3, 1, 3, 10, 20, 40, 80, 160, 320, 640, 1280, 2560 micrograms per kilogram (μg / kg).

17. The method of any one of claims 1 or 3 to 15 or the trispecific agent for use according to any one of claims 2 to 14 and 16, wherein said anti-CD19xanti-CD3xanti-CD2 trispecific agent may comprise a CD19 binding portion having CDR-H1, CDR-H2 and CDR-H3 having the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6, and CDR-L1, CDR-L2 and CDR-L3 having the amino acid sequences of SEQ ID NO: 17, SEQ ID NO: 18 and SEQ ID NO:

19.

18. 18. The method of any one of claims 15 or 17 or the trispecific agent for use according to any one of claims 16 to 17, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent comprises: (a) a first polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 37; (b) a second polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO: 38; and (c) a third polypeptide, the amino acid sequence of which comprises the amino acid sequence of SEQ ID NO:

39.

19. The method of any one of claims 1 or 3 to 15 or 17 to 18 or the trispecific agent for use of any one of claims 2 to 14 or 16 to 18, wherein the pathology or CD19-related disease or disorder is LBCL or FL3B, optionally relapsed and / or refractory LBCL or FL3B.

20. The method of any one of claims 15 or 17-18 or the trispecific agent for use of any one of claims 16-18, wherein the disease or disorder is systemic lupus erythematosus (SLE).

21. The method according to any one of claims 15 or 17 to 20 or the trispecific agent for use according to any one of claims 16 to 20, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered Q1W or Q2W.

22. The method according to any one of claims 15 or 17 to 21 or the trispecific agent for use according to any one of claims 16 to 21, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered intravenously or subcutaneously.

23. The method according to any one of claims 15 or 17 to 22 or the trispecific agent for use according to any one of claims 16 to 22, wherein the anti-CD19xanti-CD3xanti-CD2 trispecific agent may be administered by an initial priming dose followed by a main dose.

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