Therapeutic combinations including anti-CD123 immune complexes

Abstract

This provides treatment methods for hematological malignancies. [Solution] A method for treating hematological malignancies in a subject, comprising administering to a subject in need of such treatment an immune complex bound to CD123 (e.g., IMGN632), a BCL-2 inhibitor, a hypomethylating agent (e.g., azacitidine or decitabine), or a combination thereof. Methods for administering combinations to treat hematological malignancies with reduced clinical efficacy and / or toxicity are also provided. A method for treating hematological malignancies presenting as minimal residual disease using an immune complex bound to CD123 is also provided.

Description

[Technical Field]

[0001] The field of the present invention generally relates to the combination of anti-CD123 immune complexes with hypomethylating agents (HMAs) and / or B-cell leukemia / lymphoma-2 (BCL-2) antagonists, as a combination use in the treatment of hematological malignancies. [Background technology]

[0002] Cancer is one of the leading causes of death in developed countries, with more than one million people diagnosed with cancer and 500,000 dying from it each year in the United States alone. Overall, it is estimated that more than one in three people will develop some form of cancer in their lifetime.

[0003] CD123 is the alpha subunit (IL-3Rα) of the interleukin-3 receptor. CD123 expression is low in normal hematopoietic stem cells (Testa et al., Biomark Res., 10;2(1):4. (2014), Jordan et al., Leukemia, 14(10):1777-84 (2000)). However, CD123 is overexpressed in several hematological malignancies with both myeloid and lymphoid origins, including acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia in the acute transformation / acute transformation phase (BP-CML), and blastic plasmacytoid dendritic cell neoplasms (BPDCN) (Testa 2014). Interleukin-3 is produced by activated T lymphocytes. IL-3, along with other growth factors, stimulates and mediates the development and survival of a wide range of hematopoietic cells in the bone marrow (Testa 2014). CD123 levels on normal hematopoietic stem cells are very low, but early, common myeloid precursors express higher CD123 levels (Testa 2014, Jordan 2000). Medium to high expression of CD123 on normal tissues is limited to rare populations of leukocytes such as plasmacytoid dendritic cells and basophils (Jordan 2000, Testa 2014).

[0004] Acute myeloid leukemia (AML) is the most common form of acute leukemia among adults and is the leading cause of leukemia-related deaths in the United States. In 2017, an estimated 21,380 people were diagnosed with AML annually, and 10,590 patients were expected to die from the disease (Siegel et al., CA Cancer J Clin. 2017;67(1):7-30(2017)). The median age at diagnosis is 66 years. Frontline chemotherapy in AML has been reported to induce complete response (CR) in 70-80% of patients under 60 years of age and in approximately 50% of older patients. "Matched" patients are judged to be able to tolerate intensive care, are often young (under 60 years of age), and typically receive 1-2 cycles of induction with a "7+3" combination of cytarabine and anthracycline, typically daunorubicin. Following this, these matched patients may receive high-dose cytarabine for one or more cycles and then undergo stem cell transplantation. Standard induction and post-induction therapy results in a median remission period of approximately one year and a potential cure rate of 25%–35% of patients. Often elderly “unsuitable” patients typically receive venetoclax, azacitidine, and hypomethylating agents. The majority of AML patients eventually relapse, and AML salvage regimens offer poor outcomes with significant toxicity. Therefore, there is a need for novel, limited-toxicity therapies in this relapsing population.

[0005] Blastic plasmacytoid dendritic cell neoplasms (BPDCNs) are rare, invasive hematological malignancies derived from myeloid dendritic cell precursors, often presenting with cutaneous lesions in addition to lymph node, hematopoietic, and bone marrow involvement. Among other markers, BPDCN blasts are characterized by the expression of CD4, CD56, and CD123, with high levels of CD123 expression. Unfortunately, there is no standard treatment for BPDCN, and both acute lymphoblastic leukemia (ALL) and AML regimens are used as first-line treatment. Despite a complete response rate of 47%–86% in first-line disease, the median overall survival is approximately 12–16 months. The majority of BPDCN patients eventually relapse without standard treatment options.

[0006] Acute lymphoblastic leukemia (ALL) is a rare, invasive hematological malignancy originating from lymphoid precursors, often involving lymph nodes, blood, and bone marrow. B-cell ALL and some T-cell ALL blasts express CD123 at similar levels to AML blasts. While initial remission rates are high, long-term survival rates are 35-40% for patients under 60 years of age and less than 10% for elderly patients (Goldstone 2008). Several chemotherapy options are available for patients with relapsed ALL, including immunotherapy with anti-CD19 bispecific blinatumomab, which is approved by the U.S. Food and Drug Administration. However, long-term survival rates for these patients remain low. Given that currently available treatments cannot treat many hematological malignancies, more effective interventions are needed. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Testa et al.Biomark Res.,10;2(1):4.(2014) [Non-Patent Document 2] Jordan et al., Leukemia, 14(10):1777-84(2000) [Non-Patent Document 3] Siegel et al.,CA Cancer J Clin.2017;67(1):7-30(2017) [Overview of the Initiative] [Means for solving the problem]

[0008] Combinations of anti-CD123 immune complexes (e.g., IMGN632) with BCL-2 inhibitors and / or hypomethylating agents are provided herein. Methods for treating cancer patients using such combinations are also provided herein.

[0009] In certain cases, the treatment method for hematological malignancies in a subject involves administering to the subject in need an immune complex that binds to CD123 (this immune complex includes an antibody or its antigen-binding fragment, which includes a heavy chain variable region CDR1 containing the amino acid sequence of SEQ ID NO: 5, a heavy chain variable region CDR2 containing the amino acid sequence of SEQ ID NO: 6, a heavy chain variable region CDR3 containing the amino acid sequence of SEQ ID NO: 7, a light chain variable region CDR1 containing the amino acid sequence of SEQ ID NO: 8, a light chain variable region CDR2 containing the amino acid sequence of SEQ ID NO: 9, and a light chain variable region CDR3 containing the amino acid sequence of SEQ ID NO: 10), along with a BCL-2 inhibitor, a hypomethylating agent, or a combination thereof. In certain cases, the immune complex is administered in combination with a BCL-2 inhibitor. In certain cases, the immune complex is administered in combination with a hypomethylating agent. In certain cases, the immune complex is administered in combination with a BCL-2 inhibitor and a hypomethylating agent.

[0010] In certain cases, the antibody or antigen-binding fragment includes a VH containing the amino acid sequence shown in SEQ ID NO: 1, and / or a VL containing the amino acid sequence shown in SEQ ID NO: 2. In certain cases, the antibody or antigen-binding fragment includes a heavy chain constant region and / or a light chain constant region. In certain cases, the antibody or antigen-binding fragment includes a heavy chain containing the amino acid sequence shown in SEQ ID NO: 3, and / or a light chain containing the amino acid sequence shown in SEQ ID NO: 4.

[0011] In certain cases, the immune complex contains a cytotoxin. In certain cases, the cytotoxin is a DNA alkylating agent. In certain cases, the DNA alkylating agent is an indolino-benzodiazepine (IGN) DNA alkylating agent. In certain cases, the immune complex contains a peptide linker. In certain cases, the immune complex is IMGN632. In certain cases, the immune complex is administered in a pharmaceutical composition containing an immune complex having the following structure: [ka] G4723A contains a heavy chain with the amino acid sequence shown in SEQ ID NO: 3 and a light chain with the amino acid sequence shown in SEQ ID NO: 4.

[0012] In certain cases, administration is a frontline therapy. In certain cases, immune complexes are administered intravenously.

[0013] In certain cases, administration of immune complexes with BCL-2 inhibitors, hypomethylating agents, or combinations thereof produces a synergistic effect. In certain cases, administration of immune complexes and BCL-2 inhibitors does not produce more toxicity than administration of immune complexes alone or BCL-2 inhibitors alone. In certain cases, administration of immune complexes and hypomethylating agents does not produce more toxicity than administration of immune complexes alone or hypomethylating agents alone. In certain cases, administration of immune complexes, BCL-2 inhibitors, and hypomethylating agents does not produce more toxicity than administration of immune complexes, BCL-2 inhibitors, and / or hypomethylating agents.

[0014] In certain cases, the immune complex is administered once every 21 days. In certain cases, the immune complex is administered once every 21 days at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg. In certain cases, the immune complex is administered once every 21 days at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg. In certain cases, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg. In certain cases, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.135 mg / kg. In certain cases, the dose is approximately 0.045 mg / kg. In certain cases, the dose is approximately 0.03 mg / kg.

[0015] In certain cases, the immune complex is administered three times in a 21-day cycle. In certain cases, the total amount of immune complex administered in a 21-day cycle is approximately 0.045 mg / kg, approximately 0.09 mg / kg, or approximately 0.18 mg / kg. In certain cases, the total amount of immune complex administered in a 21-day cycle is approximately 0.135 mg / kg. In certain cases, an immune complex of approximately 0.015 mg / kg to approximately 0.06 mg / kg is administered in each of the three doses during the 21-day cycle. In certain cases, approximately 0.015 mg / kg, approximately 0.03 mg / kg, or approximately 0.06 mg / kg is administered in each of the three doses during the 21-day cycle. In certain cases, approximately 0.045 mg / kg is administered in each of the three doses during the 21-day cycle. In certain cases, the first dose of the immune complex is on day 7 of the 21-day cycle. In certain cases, the second dose of the immune complex is administered on day 10 of the 21-day cycle. In certain cases, the third dose is administered on day 14 of the 21-day cycle. In certain cases, the first, second, and third doses are administered on days 7, 10, and 14 of the 21-day cycle, respectively.

[0016] In certain cases, the BCL-2 inhibitor is venetoclax. In certain cases, the BCL-2 inhibitor is administered at a dose of 400 mg. In certain cases, the BCL-2 inhibitor is venetoclax. In certain cases, the BCL-2 inhibitor is administered at a dose of 200 mg.

[0017] In certain cases, the BCL-2 inhibitor is administered daily in a 21-day cycle. In certain cases, the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of a 21-day cycle, 200 mg on day 2 of a 21-day cycle, and 400 mg on days 3-21 of a 21-day cycle. In certain cases, the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of a 21-day cycle, 200 mg on day 2 of a 21-day cycle, and 400 mg on days 3-7 or 3-14 of a 21-day cycle. In certain cases, the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of a 21-day cycle, and 200 mg on days 2-21, 2-14, or 2-7 of a 21-day cycle. In certain cases, the BCL-2 inhibitor is administered on days 1-7 of a 21-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-8 of a 21-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-14 of a 21-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-18 of a 21-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-21 of a 21-day cycle. In certain cases, BCL-2 inhibitors are administered orally.

[0018] In certain cases, the administration of immune complexes is initiated 6 days after the start of BCL-2 inhibitor administration.

[0019] In certain cases, the immune complex is administered once every 28 days. In certain cases, the immune complex is administered once every 28 days at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg. In certain cases, the immune complex is administered once every 28 days at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg. In certain cases, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg. In certain cases, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.135 mg / kg. In certain cases, the dose is approximately 0.045 mg / kg. In certain cases, the dose is approximately 0.03 mg / kg.

[0020] In certain cases, the immune complex is administered three times in a 28-day cycle. In certain cases, the total amount of immune complex administered in a 28-day cycle is approximately 0.045 mg / kg, approximately 0.09 mg / kg, or approximately 0.18 mg / kg. In certain cases, the total amount of immune complex administered in a 28-day cycle is approximately 0.135 mg / kg. In certain cases, an immune complex of approximately 0.015 mg / kg to approximately 0.06 mg / kg is administered in each of the three doses during the 28-day cycle. In certain cases, an immune complex of approximately 0.015 mg / kg, approximately 0.03 mg / kg, or approximately 0.06 mg / kg is administered in each of the three doses during the 28-day cycle. In certain cases, an immune complex of approximately 0.135 mg / kg is administered in each of the three doses during the 28-day cycle. In certain cases, the first dose of the immune complex is on day 7 of the 28-day cycle. In certain cases, the second dose of the immune complex is administered on day 10 of the 28-day cycle. In certain cases, the third dose of the immune complex is administered on day 14 of the 28-day cycle. In certain cases, the first, second, and third doses of the immune complex are administered on days 7, 10, and 14 of the 28-day cycle, respectively.

[0021] In certain cases, the BCL-2 inhibitor is venetoclax. In certain cases, the BCL-2 inhibitor is administered at a dose of 400 mg. In certain cases, the BCL-2 inhibitor is administered at a dose of 200 mg.

[0022] In certain cases, the BCL-2 inhibitor is administered daily in a 28-day cycle. In certain cases, the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 28-day cycle, 200 mg on day 2 of the 28-day cycle, and 400 mg on days 3-28 of the 28-day cycle. In certain cases, the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 28-day cycle, 200 mg on day 2 of the 28-day cycle, and 400 mg on days 3-7 or 3-14 of the 28-day cycle. In certain cases, the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 28-day cycle, and 200 mg on days 2-28, 2-14, or 2-7 of the 28-day cycle. In certain cases, the BCL-2 inhibitor is administered on days 1-7 of the 28-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-8 of a 28-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-14 of a 28-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-18 of a 28-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-21 of a 28-day cycle. In certain cases, BCL-2 inhibitors are administered on days 1-28 of a 21-day cycle.

[0023] In certain cases, BCL-2 inhibitors are administered orally. In certain cases, immune complex administration is initiated 6 days after the start of BCL-2 inhibitor administration.

[0024] In certain cases, the hypomethylating agent is azacitidine. In certain cases, azacitidine is administered in a 28-day cycle. In certain cases, azacitidine is administered once daily on days 1-7 of a 28-day cycle. In certain cases, azacitidine is administered once daily on days 3-7 of a 28-day cycle. In certain cases, azacitidine is administered once daily on days 1-5 of a 28-day cycle. In certain cases, the azacitidine dose is approximately 75 mg / m². 2 It is administered in the following doses. In certain cases, azacitidine is administered subcutaneously. In certain cases, azacitidine is administered intravenously.

[0025] In certain cases, the demethylating agent is decitabine. In certain cases, decitabine is administered intravenously.

[0026] In certain cases, hematological malignancies are included as targets for minimal residual disease (MRD).

[0027] In certain cases, the treatment of hematological malignancies present as minimal residual disease in human subjects involves administering anti-CD123 immune complexes containing anti-CD123 antibodies or antigen-binding fragments linked to cytotoxic agents to the subjects. In certain cases, the immune complexes are administered at doses of approximately 0.045 mg / kg to approximately 0.18 mg / kg. In certain cases, approximately 0.045 mg / kg, approximately 0.09 mg / kg, approximately 0.135 mg / kg, or approximately 0.18 mg / kg are administered to the subjects. In certain cases, approximately 0.045 mg / kg is administered to the subjects. In certain cases, approximately 0.03 mg / kg is administered to the subjects. In certain cases, the immune complexes are administered intravenously. In certain cases, the hematological malignancies are leukemia. In certain cases, the immune complexes are administered to the subjects once every 21 days.

[0028] In certain cases, the administration is one cycle. In certain cases, the administration is two or more cycles. In certain cases, the administration is at least two cycles, at least three cycles, at least four cycles, at least five cycles, at least six cycles, at least seven cycles, at least eight cycles, at least nine cycles, or at least ten cycles. In certain cases, the administration is approximately two to four cycles, approximately two to six cycles, approximately two to eight cycles, approximately two to ten cycles, or approximately two to twelve cycles.

[0029] In certain cases, the method further includes administering a reduced dose of the immune complex after dose-limiting toxicity has occurred in the subject and been reduced to baseline or grade 2 or lower.

[0030] In certain cases, immune complexes are administered further as maintenance therapy.

[0031] In certain cases, maintenance therapy includes administering the immune complex once every 21 days. In certain cases, maintenance therapy includes administering the immune complex once every 21 days at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg. In certain cases, maintenance therapy includes administering the immune complex once every 21 days at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg. In certain cases, maintenance therapy includes administering the immune complex once every 21 days at a dose of 0.015 mg / kg. In certain cases, maintenance therapy includes administering the immune complex once every 21 days at a dose of approximately 0.045 mg / kg. In certain cases, maintenance therapy includes administering the immune complex once every 21 days at a dose of approximately 0.03 mg / kg. In certain cases, maintenance therapy includes administering immune complexes at a dose of approximately 0.09 mg / kg once every 21-day cycle. In certain cases, maintenance therapy includes administering immune complexes at a dose of approximately 0.135 mg / kg once every 21-day cycle. In certain cases, maintenance therapy includes administering immune complexes three times every 21-day cycle. In certain cases, the total amount of immune complexes administered during maintenance therapy in a 21-day cycle is approximately 0.045 mg / kg, approximately 0.09 mg / kg, or approximately 0.18 mg / kg. In certain cases, the total amount of immune complexes administered during maintenance therapy in a 21-day cycle is approximately 0.135 mg / kg. In certain cases, during maintenance therapy, immune complexes ranging from approximately 0.015 mg / kg to approximately 0.06 mg / kg are administered at each of the three doses in the 21-day cycle. In certain cases, approximately 0.015 mg / kg of immune complex is administered during maintenance therapy at each of the three 21-day cycles. In certain cases, approximately 0.03 mg / kg of immune complex is administered during maintenance therapy at each of the three 21-day cycles. In certain cases, approximately 0.045 mg / kg of immune complex is administered during maintenance therapy at each of the three 21-day cycles. In certain cases, approximately 0.06 mg / kg of immune complex is administered during maintenance therapy at each of the three 21-day cycles.

[0032] In certain cases, hematological malignancies are recurrent hematological malignancies. In certain cases, hematological malignancies are refractory hematological malignancies. In certain cases, hematological malignancies are acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia in acute transformation / acute transformation phase (BP-CML), and blastic plasmacytoid dendritic cell neoplasms (BPDCN). In certain cases, hematological malignancies are AML. In certain cases, AML is recurrent AML. In certain cases, AML is refractory AML. In certain cases, hematological malignancies are BPDCN. In certain cases, BPDCN is recurrent BPDCN. In certain cases, BPDCN is recurrent BPDCN. In certain cases, hematological malignancies are ALL. In certain cases, ALL is recurrent ALL. In certain cases, ALL is refractory ALL. In certain cases, the hematological malignancy is chronic myelomonocytic leukemia (CMML). In certain cases, CMML is relapsed CMML. In certain cases, CMML is refractory CMML. In certain cases, the hematological malignancy is myelofibrosis (MF). In certain cases, MF is relapsed MF. In certain cases, MF is refractory MF. In certain cases, the hematological malignancy is myelodysplastic syndrome (MDS). In certain cases, MDS is relapsed MDS. In certain cases, MDS is refractory MDS.

[0033] In certain cases, the hematological malignancies are those that express CD123. In certain cases, CD123 is detected in samples obtained from hematological malignancies before administration. In certain cases, CD123 is detected using flow cytometry. In certain cases, the method further includes detecting CD123 in samples obtained from hematological malignancies before administration.

[0034] In certain cases, at least 80% of cells in hematological malignancies express CD123. In certain cases, CD123 is detected in at least 80% of cells in samples obtained from hematological malignancies before administration. In certain cases, the method further includes detecting CD123 in at least 80% of cells in samples obtained from hematological malignancies before administration.

[0035] In certain cases, hematological malignancies are resistant to IMGN632.

[0036] In certain cases, hematological malignancies exhibit multidrug resistance 1 (MDR1). In certain cases, hematological malignancies express P-glycoprotein (P-gp). In certain cases, subjects have an absolute neutrophil count greater than 500 / μL.

[0037] In certain cases, the subject received at least one prior therapy. In certain cases, the subject received at least two prior therapies. In certain cases, the subject received at least three prior therapies. In certain cases, the cancer had been previously treated with venetoclax. In certain cases, the cancer had not been previously treated with venetoclax. In certain cases, the cancer had been previously treated with a hypomethylating agent. In certain cases, the cancer had not been previously treated with a hypomethylating agent.

[0038] In certain cases, subjects are pre-treated with corticosteroids before administration of the immune complex. In some cases, the method further includes pre-treating subjects with corticosteroids before administration of the immune complex. In certain cases, the corticosteroids are diphenhydramine, acetaminophen, paracetamol, dexamethasone, or a combination thereof.

[0039] In certain cases, immune complexes and BCL-2 inhibitors, hypomethylating agents, or combinations thereof are administered in separate pharmaceutical compositions.

[0040] In certain cases, immune complexes and BCL-2 inhibitors, hypomethylating agents, or combinations thereof are administered in separate pharmaceutical compositions.

[0041] In certain cases, the subject is human.

[0042] In a specific case, the treatment of hematological malignancies in human subjects involves administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg on day 7 of the cycle (optional, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg), and venetoclax administered orally at a dose of 100 mg on day 1 of the cycle, 200 mg on day 2 of the cycle, and 400 mg on days 3 to 21 of the cycle.

[0043] In a specific case, the treatment of hematological malignancies in human subjects involves administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg on day 7 of the cycle (optional, with a dose of approximately 0.135 mg / kg), and venetoclax administered orally at a dose of 100 mg on day 1 of the cycle, 200 mg on day 2 of the cycle, and 400 mg on days 3 to 21 of the cycle.

[0044] In a specific case, the treatment of hematological malignancies in human subjects involves administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously on day 7 of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (optional, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg), and venetoclax administered orally at a daily dose of 400 mg.

[0045] In certain cases, the treatment of hematological malignancies in human subjects involves administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg on day 7 of the cycle, and venetoclax administered orally at a daily dose of 400 mg. In certain cases, venetoclax is administered on days 1-7 of the 21-day cycle. In certain cases, venetoclax is administered on days 1-8 of the 21-day cycle. In certain cases, venetoclax is administered on days 1-14 of the 21-day cycle. In certain cases, venetoclax is administered on days 1-18 of the 21-day cycle. In certain cases, venetoclax is administered on days 1-21 of the 21-day cycle. In certain cases, IMGN632 is administered at a dose of approximately 0.045 mg / kg. In certain cases, IMGN632 is administered at a dose of approximately 0.03 mg / kg. In certain cases, IMGN632 is administered at a dose of approximately 0.015 mg / kg.

[0046] In a specific case, the treatment of hematological malignancies in human subjects involves administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously on day 7 of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg (optional, with a dose of approximately 0.135 mg / kg), and venetoclax administered orally at a daily dose of 400 mg.

[0047] In a specific case, the treatment of hematological malignancies in human subjects involves administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously at doses of approximately 0.015 mg / kg to approximately 0.06 mg / kg on days 7, 10, and 14 of the cycle (optionally, the doses are approximately 0.015 mg / kg, approximately 0.03 mg / kg, or approximately 0.06 mg / kg), and venetoclax administered orally at a dose of 100 mg on day 1 of the cycle, 200 mg on day 2 of the cycle, and 400 mg on days 3 through 21 of the cycle.

[0048] In a specific case, the treatment of hematological malignancies in human subjects involves administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously at doses of approximately 0.015 mg / kg to approximately 0.06 mg / kg on days 7, 10, and 14 of the cycle (optional, the doses are approximately 0.015 mg / kg, approximately 0.03 mg / kg, or approximately 0.06 mg / kg), and venetoclax administered orally at a daily dose of 400 mg.

[0049] In a specific case, the treatment of hematological malignancies in human subjects involves administering IMGN632 and azacitidine in a 28-day cycle to subjects in need, with IMGN632 administered intravenously on day 7 of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (optional, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg), and azacitidine at 75 mg / m² on days 1-7 of the cycle. 2 It is administered subcutaneously or intravenously in the specified dose.

[0050] In a specific case, the treatment method for hematological malignancies in human subjects involves administering IMGN632 and azacitidine in a 28-day cycle to subjects in need, with IMGN632 administered intravenously on day 7 of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (optional, with doses of approximately 0.015 mg / kg, 0.03 mg / kg, 0.045 mg / kg, or 0.09 mg / kg), and azacitidine at 75 mg / m². 2 It is administered subcutaneously or intravenously at the specified dose. In certain cases, azacitidine is administered on days 1–7 of the cycle. In certain cases, azacitidine is administered on days 1–5 of the cycle. In certain cases, IMGN632 is administered at a dose of approximately 0.045 mg / kg. In certain cases, IMGN632 is administered at a dose of approximately 0.03 mg / kg.

[0051] In a specific case, the treatment method for hematological malignancies in human subjects involves administering IMGN632 and azacitidine in a 28-day cycle to subjects in need, with IMGN632 administered intravenously at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg on day 7 of the cycle (optional, with a dose of approximately 0.135 mg / kg), and azacitidine administered at 75 mg / m² on days 1 to 7 of the cycle. 2 It is administered subcutaneously or intravenously in the specified dose.

[0052] In a specific case, the treatment of hematological malignancies in human subjects involves administering IMGN632 and azacitidine in a 28-day cycle to subjects in need, with IMGN632 administered intravenously at doses of approximately 0.015 mg / kg to approximately 0.06 mg / kg on days 7, 10, and 14 of the cycle (optional, with doses of approximately 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg), and azacitidine at 75 mg / m² on days 1 to 7 of the cycle. 2 It is administered subcutaneously or intravenously in the specified dose.

[0053] In a specific case, the treatment method for hematological malignancies in human subjects involves administering IMGN632 and azacitidine in a 28-day cycle to subjects in need, with IMGN632 administered intravenously at doses of approximately 0.015 mg / kg to approximately 0.06 mg / kg on days 7, 10, and 14 of the cycle, and azacitidine at 75 mg / m². 2 It is administered subcutaneously or intravenously at the specified dose. In certain cases, azacitidine is administered on days 1–7 of the cycle. In certain cases, azacitidine is administered on days 1–5 of the cycle. In certain cases, IMGN632 is administered at a dose of approximately 0.045 mg / kg. In certain cases, IMGN632 is administered at a dose of approximately 0.03 mg / kg.

[0054] In certain cases, this method further includes administering venetoclax. In certain cases, venetoclax is administered on days 1–7 of a 28-day cycle. In certain cases, venetoclax is administered on days 1–8 of a 28-day cycle. In certain cases, venetoclax is administered on days 1–14 of a 28-day cycle. In certain cases, venetoclax is administered on days 1–18 of a 28-day cycle. In certain cases, venetoclax is administered on days 1–21 of a 28-day cycle. In certain cases, venetoclax is administered on days 1–28 of a 28-day cycle.

[0055] In certain cases, the method further includes orally administering venetoclax at a dose of 100 mg on day 1 of the cycle, 200 mg on day 2 of the cycle, and 400 mg on days 3 through 28 of the cycle. In certain cases, the method further includes administering venetoclax at an oral daily dose of 400 mg.

[0056] In certain cases, the hematological malignancy is AML. In certain cases, the hematological malignancy is BPDCN. In certain cases, the hematological malignancy is ALL. In certain cases, the hematological malignancy is chronic myelomonocytic leukemia (CMML). In certain cases, the hematological malignancy is myelofibrosis (MF). In certain cases, the hematological malignancy is myelodysplastic syndrome (MDS). [Brief explanation of the drawing]

[0057] [Figure 1A] The chemical structure of IMGN632 is shown. IMGN632 is a composition containing an immune complex containing an anti-CD123 G4723A antibody linked to a cytotoxic payload DGN549-C in sodium bisulfite. The majority of the immune complexes in the composition are the sulfonated versions shown in Figure 1A. [Figure 1B] This shows a non-sulfonated form of an immune complex containing an anti-CD123 G4723A antibody linked to the cytotoxic payload DGN549-C (monoimine structure), which may also be present in the IMGN632 composition. [Figure 2A] This study demonstrates broad dose-range antitumor activity of IMGN632 against three acute myeloid leukemia (AML) cell lines (EOL-1, MV4-11, and KG-1). [Figure 2B] This study demonstrates the broad dose range of venetoclax antitumor activity against three AML (EOL-1, MV4-11, KG-1) cell lines. [Figure 2C] This demonstrates the narrower dose range of antitumor activity of IMGN632 against four AML (EOL-1, MV4-11, KG-1, and Molm-13) cell lines. [Figure 2D] This demonstrates the narrower dose range of antitumor activity of venetoclax against four AML (EOL-1, MV4-11, KG-1, and Molm-13) cell lines. [Figure 3A] The antitumor activity of IMGN632 against EOL-1 cell lines and the escalating doses of venetoclax (0.625 nM, 1.25 nM, 2.5 nM, 5 nM, 10 nM, 20 nM) are shown. [Figure 3B] The antitumor activity of IMGN632 and the escalating doses of venetoclax (0.625 nM, 1.25 nM, 2.5 nM, 5 nM, 10 nM, 20 nM) in the MV4-11 cell line are shown. [Figure 3C] The antitumor activity of IMGN632 and the escalating doses of venetoclax (0.625 nM, 1.25 nM, 2.5 nM, 5 nM, 10 nM, 20 nM) in the KG-1 cell line are shown. [Figure 3D] The antitumor activity of IMGN632 and the escalating doses of venetoclax (0.625 nM, 1.25 nM, 2.5 nM, 5 nM, 10 nM, 20 nM) in the Molm-13 cell line are shown. [Figure 4A] The efficacy of the combination of IMGN632 and venetoclax is read out as a function of dose in the EOL-1 cell line. [Figure 4B] The efficacy of the combination of IMGN632 and venetoclax is read out as a function of therapeutic effect in the EOL-1 cell line. [Figure 4C] This shows the readout of the combined efficacy of IMGN632 and venetoclax compared to the expected dose-response at a normalized scale in EOL-1 cell lines. [Figure 4D] The efficacy of the combination of IMGN632 and venetoclax is read out as a function of dose in the MV4-11 cell line. [Figure 4E] The efficacy of the combination of IMGN632 and venetoclax is read out as a function of therapeutic effect in the MV4-11 cell line. [Figure 4F] This shows the readout of the combined efficacy of IMGN632 and venetoclax compared to the expected dose-response at a normalized scale in the MV4-11 cell line. [Figure 4G] The efficacy of the combination of IMGN632 and venetoclax is read out as a function of dose in the KG-1 cell line. [Figure 4H]The efficacy of the combination of IMGN632 and venetoclax is read out as a function of therapeutic effect in the KG-1 cell line. [Figure 4I] This shows the readout of the combined efficacy of IMGN632 and venetoclax compared to the expected dose-response at a normalized scale in the KG-1 cell line. [Figure 4J] The efficacy of the combination of IMGN632 and venetoclax is read out as a function of dose in the Molm-13 cell line. [Figure 4K] The efficacy of the combination of IMGN632 and venetoclax is read out as a function of therapeutic effect in the Molm-13 cell line. [Figure 4L] This shows the readout of the combined efficacy of IMGN632 and venetoclax compared to the expected dose-response at a normalized scale in Molm-13 cell lines. [Figure 5] The in vivo efficacy of single doses of IMGN632 alone (1 μg / kg or 0.5 μg / kg), venetoclax alone (QD × 28, 100 mg / kg), and the combination of IMGN632 (1 μg / kg or 0.5 μg / kg) and venetoclax (QD × 28, 100 mg / kg) is shown by plotting median tumor volume (mm3) as a function of the number of days after inoculation with EOL-1 cell lines. [Figure 6] The in vivo efficacy of IMGN632 alone (QW×3, 3 μg / kg or 10 μg / kg), venetoclax alone (QD×28, 100 mg / kg), and the combination of IMGN632 (QW×3, 3 μg / kg or 10 μg / kg) and venetoclax (QD×28, 100 mg / kg) is shown by plotting the median tumor volume (mm3) as a function of the number of days after inoculation with the KG-1 cell line. [Figure 7]Mouse survival is shown as a function of time after inoculation with Molm-3 tumor cells. Mice received either IMGN632 alone (QW×3, 0.3 μg / kg), azacitidine alone (QD×5, 3.5 mg / kg), or a combination of IMGN632 (QW×3, 0.3 μg / kg) and azacitidine (QD×5, 3.5 mg / kg) according to three different dosing schedules. [Figure 8] Mouse survival is shown as a function of time after inoculation with Molm-3 tumor cells. Mice received either IMGN632 alone (QW×3, 0.3 μg / kg), decitabine alone (QD×5, 0.75 mg / kg), or a combination of IMGN632 (QW×3, 0.3 μg / kg) and decitabine (QD×5, 0.75 mg / kg) according to three different dosing schedules. [Figure 9] The in vivo efficacy of single doses of IMGN632 alone (1 μg / kg), venetoclax (QD × 28, 100 mg / kg), and azacitidine (QD × 5, 3 mg / kg), or IMGN632 (1 μg / kg) in combination with venetoclax (QD × 28, 100 mg / kg) and azacitidine (QD × 5, 3 mg / kg), is shown by plotting median tumor volume (mm3) as a function of the number of days after inoculation of EOL-1 cell lines. [Figure 10A] Mouse survival is shown as a function of time after inoculation with Molm-3 tumor cells. Mice received IMGN632 alone (QW×3), venetoclax (QD×28), and azacitidine (QD×5), or IMGN632 (QW×3) in combination with venetoclax (QD×28) and azacitidine (QD×5). [Figure 10B] Mouse survival is shown as a function of time after inoculation with Molm-3 tumor cells. Mice received IMGN632 alone (QW×3), venetoclax (QD×28), and azacitidine (QD×5), or IMGN632 (QW×3) in combination with venetoclax (QD×28) and azacitidine (QD×5). [Figure 11]Mouse survival is shown as a function of time after inoculation with MV4-11 tumor cells. Mice received IMGN632 alone (QW×3), venetoclax (QD×28), and azacitidine (QD×5), or IMGN632 (QW×3) in combination with venetoclax (QD×28) and azacitidine (QD×5). [Modes for carrying out the invention]

[0058] The present invention provides anti-CD123 immune complexes in combination with hypomethylating agents (HMAs) and / or B-cell leukemia / lymphoma-2 (BCL-2) antagonists, and the use of such combinations in the treatment of hematological malignancies. The present invention also provides anti-CD123 immune complexes (optionally in combination with HMAs and / or BCL-2 antagonists) for the treatment of hematological malignancies presenting as minimal residual disease.

[0059] I. Definition To facilitate understanding of the present invention, several terms and phrases are defined below.

[0060] The terms “IL-3Rα,” “interleukin-3 receptor α,” and “CD123,” as used interchangeably herein, refer to mammalian CD123 polypeptides, including but not limited to the naturally occurring CD123 polypeptide and its isoforms, unless otherwise indicated. These terms encompass the “full-length” unprocessed CD123 polypeptide and any form of CD123 polypeptide resulting from intracellular processing. The terms also encompass naturally occurring variants of CD123, such as those encoded by splice variants and allele variants. The CD123 polypeptides described herein can be isolated from a variety of sources, such as human tissue types or other sources, or prepared by recombinant or synthetic methods. Where specifically indicated, “CD123” may be used to refer to the nucleic acid encoding the CD123 polypeptide. Human CD123 sequences are known and include, for example, sequences associated with NCBI reference numbers NP_002174 and NM_002183 (protein and nucleic acid sequences of human CD123 variant 1), and sequences associated with NP_001254642 and NM_001267713 (protein and nucleic acid sequences of human CD123 variant 2). As used herein, the term "human CD123" refers to CD123 containing the sequence of SEQ ID NO: 11 or SEQ ID NO: 12. MVLLWLTLLL IALPCLLQTK EDPNPPITNL RMKAKAQQLT WDLNRNVTDI ECVKDADYSM PAVNNSYCQF GAISLCEVTN YTVRVANPPF STWILFPENS GKPWAGAENL TCWIHDVDFL SCSWAVGPGA PADVQYDLYL NVANRRQQYE CLHYKTDAQG TRIGCRFDDI SRLSSGSQSS HILVRGRSAA FGIPCTDKFV VFSQIEILTP PNMTAKCNKT HSFMHWKMRS HFNRKFRYEL QIQKRMQPVI TEQVRDRTSF QLLNPGTYTV QIRARERVYE FLSAWSTPQR FECDQEEGAN TRAWRTSLLI ALGTLLALVC VFVICRRYLV MQRLFPRIPH MKDPIGDSFQ NDKLVVWEAG KAGLEECLVT EVQVVQKT(Sequence ID 11) MVLLWLTLLL IALPCLLQTK EGGKPWAGAE NLTCWIHDVD FLSCSWAVGP GAPADVQYDL YLNVANRRQQ YECLHYKTDA QGTRIGCRFD DISRLSSGSQ SSHILVRGRS AAFGIPCTDK FVVFSQIEIL TPPNMTAKCN KTHSFMHWKM RSHFNRKFRY ELQIQKRMQP VITEQVRDRT SFQLLNPGTY TVQIRARERV YEFLSAWSTP QRFECDQEEG ANTRAWRTSL LIALGTLLAL VCVFVICRRY LVMQRLFPRI PHMKDPIGDS FQNDKLVVWE AGKAGLEECL VTEVQVVQKT (array number 12)

[0061] The term “antibody” means an immunoglobulin molecule that recognizes and specifically binds to a target such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or a combination thereof, via at least one antigen recognition site within the variable region of the immunoglobulin molecule. As used herein, the term “antibody” includes intact polyclonal antibodies, intact monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, antibody-containing fusion proteins, and any other modified immunoglobulin molecules, as long as these antibodies exhibit the desired biological activity. Antibodies can be any of the five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, or their subclasses (isotypes) (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), based on the identity of their heavy chain constant domains, which are respectively referred to as α, δ, ε, γ, and μ. Different classes of immunoglobulins have different known subunit structures and three-dimensional configurations. Antibodies can be naked or conjugated with other molecules such as toxins or radioactive isotopes.

[0062] The term "anti-CD123 antibody" or "CD123-binding antibody" refers to an antibody that can bind to CD123 with sufficient affinity to be useful as a diagnostic and / or therapeutic agent when the antibody targets CD123 (e.g., an antibody in IMGN632). The degree of binding of an anti-CD123 antibody to unrelated non-CD123 proteins may be less than 10% of the binding of an antibody to CD123 as measured, for example, by radioimmunoassay (RIA).

[0063] The term "antibody fragment" refers to a portion of an intact antibody that has a sufficient positive charge to bind to a cation exchange resin. "Antigen-binding fragment" refers to a portion of an intact antibody that binds to an antigen and has a sufficient positive charge to bind to a cation exchange resin. Antigen-binding fragments may contain the antigen-determining variable region of an intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, linear antibodies, and single-chain antibodies.

[0064] The term “cysteine-manipulated” antibody or antigen-binding fragment includes an antibody or antigen-binding fragment having at least one cysteine ​​("Cys") that is not normally present in a given residue of the light or heavy chain of the antibody or antigen-binding fragment. Such a Cys, which may also be referred to as “manipulated Cys,” can be manipulated using any conventional molecular biology or recombinant technique (e.g., by replacing the coding sequence of a non-Cys residue at a target residue with the coding sequence of Cys). For example, if the original residue is a Ser having the coding sequence 5'-UCU-3', the coding sequence can be mutated (e.g., by site-directed mutagenesis) to 5'-UGU-3' which codes for Cys. In certain embodiments, the Cys-manipulated antibody or antigen-binding fragment has the manipulated Cys in the heavy chain. In certain embodiments, the manipulated Cys is in or near the CH3 domain of the heavy chain. In certain embodiments, the manipulated Cys is located at residue 442 of the heavy chain (EU / OU numbering; EU index, Kabat et al, Sequences of Proteins of Immunological Interest, 5th Ed., NIH publication No. 91-3242, 1991, its entirety incorporated herein by reference). In certain embodiments, the Fc region contains cysteine ​​at one or more of the positions 239, 282, 289, 297, 312, 324, 330, 335, 337, 339, 356, 359, 361, 383, 384, 398, 400, 440, 422, and 442, as numbered by the EU index. In certain embodiments, one or more of the following residues may be substituted with cysteine. The light chain is designated V205 (Kabat numbering), the heavy chain A118 (EU numbering), and the heavy chain Fc region S400 (EU numbering). In certain embodiments, for example, the variable light chain domain of scFv has cysteine ​​at Kabat position 100. In certain embodiments, for example, the variable heavy chain domain of scFv has cysteine ​​at Kabat position 44.Cysteine-modified antibodies may be produced, for example, as described in U.S. Patent Nos. 7,521,541, 7,855,275, U.S. Publication No. 2011 / 0033378 and WO2011 / 005481.

[0065] A “monoclonal” antibody or its antigen-binding fragment refers to a homogeneous population of antibodies or antigen-binding fragments involved in the highly specific recognition and binding of a single antigenic determinant or epitope. This is in contrast to polyclonal antibodies, which typically contain different antibodies directed toward different antigenic determinants. The term “monoclonal” antibody or its antigen-binding fragment encompasses both intact and full-length monoclonal antibodies, as well as antibody fragments (e.g., Fab, Fab', F(ab')2, Fv), single-chain (scFv) variants, fusion proteins containing the antibody moiety, and any other modified immunoglobulin molecules containing an antigen recognition site. Furthermore, “monoclonal” antibodies or their antigen-binding fragments refer to such antibodies and their antigen-binding fragments produced by any number of methods, including but not limited to hybridomas, phage selection, recombinant expression, and transgenic animals.

[0066] The term “humanized” antibody or antigen-binding fragment refers to a form of non-human (e.g., mouse) antibody or antigen-binding fragment that contains a specific immunoglobulin chain, a chimeric immunoglobulin, or a fragment thereof containing minimal non-human (e.g., mouse) sequences. Typically, a humanized antibody or antigen-binding fragment is a human immunoglobulin in which residues from the complementary determinant region (CDR) are replaced with residues from a non-human species (e.g., mouse, rat, rabbit, hamster) CDR that has the desired specificity, affinity, and ability ("implanted CDR") (Jones et al., Nature 321:522-525 (1986), Riechmann et al., Nature 332:323-327 (1988), Verhoeyen et al., Science 239:1534-1536 (1988)). In some cases, the Fv framework region (FR) residues of human immunoglobulins are replaced with corresponding residues in non-human species antibodies or fragments having the desired specificity, affinity, and capability. Humanized antibodies or their antigen-binding fragments can be further modified by substitution of additional residues within the Fv framework region and / or within the substituted non-human residues to refine and optimize the specificity, affinity, and / or capability of the antibody or its antigen-binding fragment. Generally, humanized antibodies or their antigen-binding fragments contain at least one, typically two or three, variable domains, including all or substantially all of the CDR region corresponding to the non-human immunoglobulin, but all or substantially all of the FR region is from the human immunoglobulin consensus sequence. Humanized antibodies or their antigen-binding fragments may also contain at least a portion of the immunoglobulin constant region or domain (Fc), typically from human immunoglobulin. Examples of methods used to generate humanized antibodies are described in U.S. Patent No. 5,225,539, Roguska et al., Proc. Natl. Acad. Sci., USA, 91(3):969-973 (1994), and Roguska et al., Protein Eng. 9(10):895-904 (1996). In some embodiments, the “humanized antibody” is a resurfaced antibody.

[0067] The "variable region" of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, either individually or in combination. The variable regions of the heavy and light chains each consist of four framework regions (FRs) connected by three complementarity-determining regions (CDRs), also known as hypervariable regions. The CDRs in each chain are held together in close proximity by the FRs and, together with CDRs from other chains, contribute to the formation of the antibody's antigen-binding site. There are at least two techniques for determining CDRs: (1) an approach based on interspecific sequence variability (i.e., Kabat et al., Sequences of Proteins of Immunological Interest, (5th ed., 1991, National Institutes of Health, Bethesda Md.), "Kabat"), and (2) an approach based on crystallographic studies of antigen-antibody complexes (Al-lazikani et al, J. Molec. Biol. 273:927-948 (1997)). In addition, a combination of these two approaches is sometimes used in the art to determine the CDR.

[0068] The Kabat numbering system is generally used to refer to residues within the variable domain (approximately residues 1-107 of the light chain and residues 1-113 of the heavy chain) (e.g., Kabat et al., Sequences of Immunological Interest (5th Ed., 1991, National Institutes of Health, Bethesda, Md.) ("Kabat").

[0069] Kabat amino acid reposition numbering refers to the numbering system used in heavy-chain or light-chain variable domains of antibody editing by Kabat et al. (Sequences of Immunological Interest. (5th Ed., 1991, National Institutes of Health, Bethesda, Md.), "Kabat"). Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to the shortening or insertion of FR or CDR in the variable domain. For example, a heavy-chain variable domain may include a single amino acid insertion after H2 residue 52 (residue 52a according to Kabat) and a residue inserted after heavy-chain FR residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat). The Kabat numbering of residues can be determined for a given antibody by alignment of the antibody sequence with a region of homology to a "standard" Kabat numbered sequence. Instead, Chothia refers to the location of the structural loop (Chothia and Lesk, J.Mol.Biol.196:901-917(1987)). When numbered using Kabat numbering rules, the end of the Chothia CDR-H1 loop varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places insertions at H35A and H35B; if neither 35A nor 35B exists, the loop ends at 32; if only 35A exists, the loop ends at 33; if both 35A and 35B exist, the loop ends at 34). The AbM hypervariable region represents a compromise between Kabat CDR and Chothia structural loops and is used by Oxford Molecular's AbM antibody modeling software. [Table A]

[0070] The term "human" antibody or its antigen-binding fragment means an antibody or its antigen-binding fragment produced by a human, or an antibody or its antigen-binding fragment having an amino acid sequence corresponding to a human-produced antibody or its antigen-binding fragment prepared using any technique known in the art. This definition of human antibody or its antigen-binding fragment includes intact or full-length antibodies and their fragments.

[0071] The term "chimeric" antibody or antigen-binding fragment refers to an antibody or antigen-binding fragment whose amino acid sequence originates from two or more species. Typically, both the light and heavy chain variable regions correspond to the variable regions of an antibody or antigen-binding fragment derived from a certain mammal (e.g., mouse, rat, rabbit, etc.) that possesses the desired specificity, affinity, and ability, while the constant region is homologous to the sequence of an antibody or antigen-binding fragment derived from another species (usually human) that does not induce an immune response in that species.

[0072] The terms “epitope” and “antigenic determinant” are used interchangeably herein and refer to a portion of an antigen that is recognized and specifically bound to a particular antibody. When the antigen is a polypeptide, epitopes can be formed from both sequential and discontinuous amino acids juxtaposed by the tertiary folding of the protein. Epitopes formed from sequential amino acids are typically retained during protein denaturation, while epitopes formed by tertiary folding are typically lost during protein denaturation. Epitopes typically contain at least three, more commonly at least five or eight to ten, amino acids in a unique spatial conformation.

[0073] "Binding affinity" generally refers to the strength of the combined non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects the 1:1 interaction between the members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y can generally be expressed by a dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described herein. Low-affinity antibodies generally tend to bind slowly to antigens and dissociate easily, while high-affinity antibodies generally tend to bind more quickly to antigens and remain bound for longer. Various methods for measuring binding affinity are known in the art, and any of them can be used for the purposes of this disclosure. Specific exemplary embodiments are described below.

[0074] When used herein to refer to binding affinity, "or better" refers to a stronger bond between a molecule and its binding partner. When used herein, "or better" refers to a stronger bond represented by a smaller numerical Kd value. For example, an antibody having an affinity for an antigen of "0.6 nM or greater," where the affinity of the antibody for the antigen is less than 0.6 nM, i.e., 0.59 nM, 0.58 nM, 0.57 nM, etc., or any value less than 0.6 nM.

[0075] "Specifically binding" generally means that an antibody binds to an epitope via its antigen-binding domain, and that this binding involves some degree of complementarity between the antigen-binding domain and the epitope. According to this definition, when an antibody binds to an epitope, it is said to "specifically bind" to that epitope more easily via its antigen-binding domain than it would bind to a random, unrelated epitope. The term "specificity" is used herein to modify the relative affinity with which a particular antibody binds to a particular epitope. For example, antibody "A" may be considered to have higher specificity for a given epitope than antibody "B", or antibody "A" may be said to bind to epitope "C" with higher specificity than the relevant epitope "D".

[0076] "Preferential binding" means that the antibody specifically binds to an epitope more readily than it binds to the associated, similar, homologous, or analogous epitopes. Therefore, an antibody that "preferentially binds" to a given epitope is more likely to bind to that epitope than to the associated epitope, although such an antibody may cross-react with the associated epitope.

[0077] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to polymers of amino acids of any length. Polymers may be linear or branched, may contain modified amino acids, and may be interrupted by non-amino acids. These terms also encompass naturally occurring or interveningly modified amino acid polymers, and any other operations or modifications such as disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or conjugation with labeling components. The definitions also include polypeptides containing, for example, one or more analogues of amino acids (including, for example, unnatural amino acids), as well as other modifications known in the art. Since the polypeptides of this disclosure are antibody-based, it will be understood that in certain embodiments, polypeptides may arise as single-chain or linked-chain polymers.

[0078] As used herein, the terms “immune complex” or “complex” refer to a compound or derivative thereof that is linked to a cell-binding agent (i.e., an anti-CD123 antibody or a fragment thereof) and is defined by the following general formula: CA (C = cytotoxin (e.g., indolino-benzodiazepine (IGN) DNA alkylating agent (e.g., DGN549-C)), and A = antibody or its antigen-binding fragment, e.g., anti-CD123 antibody or antibody fragment). An immune complex may optionally contain a linker and may be defined by the general formula CLA (C = cytotoxin, L = linker, and A = antibody or its antigen-binding fragment, e.g., anti-CD123 antibody or antibody fragment). An immune complex may also be defined by the general formula in the reverse order: CA or ALC. An immune complex may also contain multiple cytotoxins (C) per antibody or its antigen-binding fragment (A), or multiple cytotoxins (C) and a linker (L) per antibody or its antigen-binding fragment (A).

[0079] A "linker" is any chemical moiety that can link a compound, typically a drug (such as an IGN DNA alkylating agent), to a cell binding agent (such as an anti-CD123 antibody or a fragment thereof) in a stable, covalent manner. The linker may be sensitive to, or substantially resistant to, acid-inducible cleavage, photo-inducible cleavage, peptidase-inducible cleavage, esterase-inducible cleavage, and disulfide bond cleavage under conditions that allow the compound or antibody to maintain its activity. Suitable linkers are well known in the art and include, for example, disulfide groups, thioether groups, acid-unstable groups, photo-unstable groups, peptidase-unstable groups, and esterase-unstable groups. Linkers also include charged linkers, and their hydrophilic forms, as described herein and known in the art. In some embodiments disclosed herein, the linker is a peptide linker.

[0080] The term "IMGN632" refers to the immune complex composition shown in Figures 6A and 6B. The immune complex composition contains an immune complex containing an average of 1.5–2.1 DGN549-C cytotoxic agent per sulfonated version of huCD123-6Gv4.7 ("G4723A") antibody (Figure 1A). The immune complex composition may also contain an unsulfonated immune complex (monoimine structure shown in Figure 1B).

[0081] The term "BCL-2 inhibitor" refers to drugs that can inhibit the activity of B-cell leukemia / lymphoma-2 ("BCL-2"). For example, BCL-2 inhibitors can bind to BCL-2 and reduce the interaction between BCL-2 and pro-apoptotic proteins (e.g., the BH3-only protein). Venetoclax is an example of a BCL-2 inhibitor.

[0082] The term "hypomethylating agent" or "HMA" refers to drugs that inhibit DNA methylation. For example, HMAs can act by inhibiting DNA methyltransferase. Azacitidine and decitabine are exemplary HMAs.

[0083] The terms “cancer” and “cancerous” refer to or describe a physiological condition in mammals characterized by uncontrolled cell growth of a population of cells. Examples of cancer include, but are not limited to, cancer, lymphoma, blastoma, sarcoma, and leukemia. “Tumor” and “neoplasm” refer to one or more cells resulting from excessive cell growth or proliferation, whether benign (non-cancerous) or malignant (cancerous), including precancerous lesions. Cancers disclosed herein may be hematological malignancies. Examples of hematological malignancies include acute myeloid leukemia (AML), chronic myeloid leukemia (CML), myelodysplastic syndrome (MDS), acute lymphoblastic leukemia (ALL) such as B-cell acute lymphoblastic leukemia (B-ALL), T-cell acute lymphoblastic leukemia (T-ALL), mixed leukemia ALL (MLL-ALL), B-cell precursor ALL (BCP-ALL), Ph+ALL, Ph-like ALL, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia in the acute transformation / acute transformation phase (BP-CML), and blastic plasmacytoid dendritic cell neoplasms (BPDCN). Further examples of "cancer" include B-cell lymphomas, including NHL; mature B-cell tumors such as precursor B-cell lymphoblastic leukemia / lymphoma and B-cell chronic lymphocytic leukemia (CLL) / small lymphocytic lymphoma (SLL); B-cell prelymphocytic leukemia; lymphoplasmacytic lymphoma; mantle cell lymphoma (MCL); follicular lymphoma (FL), including low-grade, intermediate-grade, and high-grade FL; cutaneous follicular lymphoma; marginal zone B-cell lymphoma (MALT type, nodular type, and splenic type); hairy cell leukemia; diffuse large B-cell lymphoma; Burkitt lymphoma; plasmacytoma; plasmacytomyeloma; post-transplant lymphoproliferative disorder; Waldenström macroglobulinemia; and anaplastic large cell lymphoma (ALCL). Cancer may be a cancer that expresses CD123 ("CD123-expressing cancer").

[0084] The terms “cancer cells,” “tumor cells,” and grammatically equivalent terms refer to the total population of cells derived from a tumor or precancerous lesion, including both non-tumorogenic cells, which constitute the majority of the tumor cell population, and oncogenic stem cells (cancer stem cells). As used herein, the term “tumor cells” is modified by the term “non-tumorogenic” if it refers only to tumor cells that lack the ability to regenerate and differentiate in order to distinguish these tumor cells from cancer stem cells.

[0085] "Refractory" cancers are those that progress even when cancer patients are administered anti-cancer drug treatments such as chemotherapy. Cancers can be resistant at the start of treatment or can become resistant during treatment.

[0086] "Primary refractory" cancer is a type of cancer in which a patient does not achieve complete remission (CR) or complete remission with incomplete recovery (CRi) after two cycles of intensive chemotherapy.

[0087] "Recurrent" cancer is when cancer or signs and symptoms of cancer return after a period of improvement.

[0088] As used herein, the term “matched AML” refers to subjects with AML who are candidates for intensive therapy. Measures used to determine a subject with matched AML include, for example, physical performance (determined by, for example, the East Coast Cancer Clinical Trials Group Performance Status (ECOG PS), Karnovsky Performance Status (KPS), and Short Physical Performance Battery (SPPB)), comorbidities (determined by the Charlson Comorbidity Index (CCI) or Hematopoietic Stem Cell Transplant-Specific Comorbidity Index (HCT-CI)), cognitive function, and prognostic models (including, but not limited to, cytogenetic group, age, white blood cell count, LDH, and type of AML). In some cases, a matched AML subject is a subject under 60 years of age.

[0089] As used herein, the term “incompatible AML” refers to subjects with AML that are not candidates for intensive therapy. Measures used to determine subjects with incompatible AML include, for example, physical performance (determined by, for example, the East Coast Cancer Clinical Trials Group Performance Status (ECOG PS), Karnovsky Performance Status (KPS), and Short Physical Performance Battery (SPPB)), comorbidities (determined by the Charlson Comorbidity Index (CCI) or Hematopoietic Stem Cell Transplant-Specific Comorbidity Index (HCT-CI)), cognitive function, and prognostic models (including, but not limited to, cytogenetic group, age, white blood cell count, LDH, and type of AML). In some cases, subjects with incompatible AML are those over 60 years of age.

[0090] The term “increased expression” or “overexpression” of CD123 in a particular tumor, tissue, or cell sample refers to CD123 present at higher levels than that found in healthy or non-disease (natural, wild-type) tissue or cells of the same type or origin.

[0091] Terms such as “to treat,” “to cure,” “to treat,” or “to alleviate,” or “to alleviate,” refer to therapeutic measures that cure, alleviate, reduce the symptoms of, and / or halt the progression of a diagnosed pathological condition or disorder. Therefore, those who require treatment include those who have already been diagnosed with a disorder or who are suspected of having a disorder.

[0092] The term “therapeutic effective dose” refers to the amount of an antibody, immune complex, or other drug that is effective in “treating” a disease or disorder in a subject or mammal. In the case of cancer, a therapeutic effective dose of a drug can reduce the number of cancer cells, reduce tumor size or load, inhibit (i.e., slow to some extent, and in certain embodiments, halt) cancer cell infiltration into peripheral organs, alleviate to some extent one or more of the symptoms associated with cancer, and / or produce a favorable response such as complete remission (CR), complete remission with incomplete recovery (CRi), complete remission with partial hematological recovery (CRh), CR without minimal residual disease (CRMRD-), clinical complete remission (CRc), morphological non-leukemia state (MLFS), partial remission (PR), duration of response (DOR), and reduction of progressive disease (PD).

[0093] The term "favorable response" generally refers to producing a beneficial state in a subject. In the context of cancer treatment, this term refers to providing a therapeutic effect to the subject. Positive therapeutic effects in cancer can be measured in several ways (see WAWeber, J.Nucl.Med. 50:1S-10S (2009)). A favorable response may be assessed, for example, by complete remission (CR), complete remission with incomplete recovery (CRi), CR without minimal residual disease (CRMRD-), clinical complete remission (CRc), morphological leukemia-free state, partial remission (PR), reduction of progressive disease (PD), or any combination thereof.

[0094] "Complete response," "complete remission," or "CR" indicates the disappearance of all signs of a tumor or cancer in response to treatment. This does not necessarily mean that the cancer is cured. "CRi" refers to morphologically complete remission with incomplete hematological (blood cell count) recovery. "CRMRD-" refers to complete recovery with no measurable residual disease.

[0095] "Minimally Invasive Disease," "MRD," or "MRD+" refers to the persistence of cancer (e.g., leukemia) cells after treatment at levels below morphological detection. MRD can be assessed, for example, using central flow cytometry. MRD+ status is a predictor of recurrence and has been associated with reduced survival rates in AML patients.

[0096] "Partial response" or "PR" refers to a reduction in the size or volume of one or more tumors or lesions, or the extent of cancer in the body, in response to treatment.

[0097] "Progressive disease" refers to the appearance of one or more new lesions or tumors, and / or the clear progression of an existing non-target lesion. Progressive disease can also refer to tumor growth of more than 20% since the start of treatment, resulting from either an increase in tumor mass or tumor spread.

[0098] The terms “treatment” or “therapy” refer to a treatment regimen that may include, but is not limited to, surgery, radiotherapy, chemotherapy, differentiation therapy, biotherapy, immunotherapy, induction therapy, consolidation therapy, transplantation, maintenance therapy, or administration of one or more anticancer agents (e.g., cytotoxic agents and / or antiproliferative compounds).

[0099] The terms “first-line treatment,” “first-line therapy,” and “frontline therapy” refer to preferred standard initial treatments for specific conditions, such as induction therapy, intensification therapy, transplantation, and maintenance therapy. These treatments are distinct from “second-line” therapies, which are attempted when first-line therapy is ineffective. “Third-line” therapies are attempted when first-line and second-line therapies are ineffective. “Salvage therapy” or “salvage regimen” is a treatment attempted when cancer has not responded to other treatments.

[0100] For example, the combination of the CD123 immune complex (e.g., IMGN632) provided herein with a BCL-2 inhibitor and / or HMA may be given as a first-line, second-line, or third-line therapy. The combination of the CD123 immune complex (e.g., IMGN632) provided herein with a BCL-2 inhibitor and / or HMA may be given as a therapy for patients who have received at least one, at least two, or at least three therapies (e.g., a frontline therapy and one salvage therapy) prior to treatment with the combination of the CD123 immune complex (e.g., IMGN632) provided herein with a BCL-2 inhibitor and / or HMA. In some embodiments, the combination of the CD123 immune complex (e.g., IMGN632) provided herein with a BCL-2 inhibitor and / or HMA may be given as a therapy for patients who have received one or fewer, two or fewer, three or fewer, four or fewer, five or fewer, or six or fewer therapies.

[0101] The term "maintenance therapy" refers to treatment given to prevent cancer recurrence after it has disappeared following initial treatment.

[0102] The phrase "pharmaceutically acceptable" indicates that a substance or composition must be chemically and / or toxicologically compatible with the other components of the formulation and / or the mammal being treated with the formulation.

[0103] The term "pharmaceutical preparation" refers to a preparation that is in a form that allows the biological activity of the active ingredient to be effective and does not contain additional ingredients that are unacceptably toxic to the subject to which the preparation is administered. The preparation may be sterile.

[0104] The term "subject" refers to any animal (e.g., mammal) that is a recipient of a particular treatment, including but not limited to humans, non-human primates, and rodents. Typically, the terms "subject" and "patient" are used interchangeably herein in reference to human subjects.

[0105] Combination therapy with one or more additional therapeutic agents includes simultaneous (parallel) or sequential administration in any order.

[0106] Combination therapy provides a "synergistic effect," which can be demonstrated when the combined active ingredients are greater than the combined effect of using the compounds separately. A synergistic effect can be obtained when the active ingredients are (1) formulated simultaneously and administered or delivered simultaneously in a combined unit dose formulation, (2) delivered sequentially, alternately, or in parallel as separate formulations, or (3) delivered by several other regimens. In the case of alternating therapy, a synergistic effect can be obtained when the compounds are administered or delivered sequentially, for example, by different injections in separate syringes.

[0107] The term "to instruct" means to provide written instructions regarding applicable therapies, medications, treatments, treatment regimens, etc., by any means, such as in the form of package inserts or other written promotional materials.

[0108] The “effective dose” of any antibody, immune complex, or other drug disclosed herein is an amount sufficient to perform the purpose specifically described. The “effective dose” can be determined empirically and routinely in relation to the purpose described.

[0109] The polypeptides, antibodies, polynucleotides, vectors, cells, or compositions “isolated” are polypeptides, antibodies, polynucleotides, vectors, cells, or compositions in a form not found in nature. Isolated polypeptides, antibodies, polynucleotides, vectors, cells, or compositions include those purified to the extent that they are no longer found in nature. In some embodiments, the isolated antibodies, polynucleotides, vectors, cells, or compositions are substantially pure.

[0110] As used herein, “substantially pure” means a material that is at least 50% pure (i.e., free from contaminants), at least 90% pure, at least 95% pure, at least 98% pure, or at least 99% pure.

[0111] As used in this disclosure and in the claims, the singular forms "a," "an," and "the" include the plural form unless the context expressly indicates otherwise.

[0112] Where embodiments are described in this specification using the language “including,” it should be understood that similar embodiments otherwise described using the words “consisting of” and / or “essentially consisting of” are also provided.

[0113] The term "and / or" as used in phrases such as "A and / or B" in this specification is intended to include both "A and B," "A or B," and "A" and "B." Similarly, the term "and / or" as used in phrases such as "A, B, and / or C" is intended to include each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0114] II. Anti-CD123 immune complex This specification describes a method for administering an immune complex that specifically binds to CD123 (e.g., IMGN632) in combination with other agents. The immune complex that specifically binds to CD123 is referred to herein as a "CD123-immune complex" or "anti-CD123 immune complex." Such an immune complex comprises an anti-CD123 antibody or its antigen-binding fragment, and a drug (e.g., a cytotoxic agent). The drug (e.g., a cytotoxic agent) can be attached to the anti-CD123 antibody or its antigen-binding fragment by a linker.

[0115] In some embodiments, the anti-CD123 antibody or antigen-binding fragment thereof is a humanized antibody or antigen-binding fragment thereof. In some embodiments, the humanized antibody or fragment is a resurfaced antibody or antigen-binding fragment thereof. In other embodiments, the antibody or antigen-binding fragment thereof is a fully human antibody or antigen-binding fragment thereof.

[0116] In one embodiment, the immune complex is represented by the following formula:

Chemical formula

[0117] In the above formula, Cy c1 is represented by the following formula:

Chemical formula

Chemical formula

Chemical formula

[0118] In a particular embodiment, R a and R b Both are H, and R5 is either H or Me.

[0119] In certain embodiments, P is a peptide containing 2 to 5 amino acid residues. For example, P may be Gly-Gly-Gly, Ala-Val, Val-Ala, Val-Cit, Val-Lys, Phe-Lys, Lys-Lys, Ala-Lys, Phe-Cit, Leu-Cit, Ile-Cit, Trp, Cit, Phe-Ala, Phe-N 9 -Tosyl-Arg, Phe-N 9-Nitro-Arg, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Leu-Ala-Leu, Ile-Ala-Leu, Val-Ala-Val , Ala-Leu-Ala-Leu, β-Ala-Leu-Ala-Leu, Gly-Phe-Leu-Gly, Val-Arg, Arg-Val, Arg-Arg, Val-D-Ci t, Val-D-Lys, Val-D-Arg, D-Val-Cit, D-Val-Lys, D-Val-Arg, D-Val-D-Cit, D-Val-D-Lys, D-Val-D-Arg, D-Arg-D-Arg, Ala-Ala, Ala-D-Ala, D-Ala-Ala, D-Ala-D-Ala, Ala-Met, and Met-Ala. In certain embodiments, P is Gly-Gly-Gly, Ala-Val, Ala-Ala, Ala-D-Ala, D-Ala-Ala, or D-Ala-D-Ala. In certain embodiments, Q is -SO3M.

[0120] In a particular embodiment, R 19 and R 20 Both are H, and m'' is an integer between 1 and 6.

[0121] In a particular embodiment, -L c - is represented by the following formula: [ka]

[0122] In a particular embodiment, the immune complex is expressed by the following formula [ka] [ka] or represented by its pharmaceutically acceptable salt, in the formula, a double line between N and C. [ka] X represents a single bond or a double bond, however, if it is a double bond, X does not exist and Y is -H, and if it is a single bond, X is -H and Y is -OH or -SO3M.

[0123] For example, an anti-CD123 antibody or its antigen-binding fragment may be present in the immune complex used in this method. Anti-CD123 antibodies or their antigen-binding fragments are described (see, for example, U.S. Patent No. 10,077,313 B2, the contents of which are incorporated herein by reference in their entirety). The anti-CD123 antibody or its antigen-binding fragment may be a huCD123-6Gv4.7 ("G4723A") antibody (see WO2017 / 004025, WO2017 / 004026, and PCT / US2018 / 052212, the contents of which are incorporated herein by reference in their entirety), or it may contain, for example, the sequence of a G4723A antibody as shown in Tables 1-3 below. For example, an anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein may include variable heavy chains CDR-1, CDR-2, and CDR-3 containing the sequences of SEQ ID NOs. 5, 6, and 7, respectively, and / or variable light chains CDR-1, CDR-2, and CDR-3 containing the sequences of SEQ ID NOs. 8, 9, and 10, respectively. An anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein may include a variable heavy chain domain containing the sequence shown in SEQ ID NO. 1. An anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein may include a variable light chain domain containing the sequence shown in SEQ ID NO. 2. An anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein may include a variable heavy chain domain containing the sequence shown in SEQ ID NO. 2. An anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein may include a heavy chain containing the sequence shown in SEQ ID NO. 3. An anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein may include a light chain containing the sequence shown in SEQ ID NO: 4. An anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein may include a heavy chain containing the sequence shown in SEQ ID NO: 3 and a light chain containing the sequence shown in SEQ ID NO: 4. [Table 1] [Table 2] [Table 3]

[0124] An anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein can bind to an epitope within amino acids 205-346 of human CD123.

[0125] Anti-CD123 antibodies or their antigen-binding fragments for use in the methods provided herein can be produced recombinantly. For example, anti-CD123 antibodies or their antigen-binding fragments for use in the methods provided herein can be produced in mammalian cell lines, such as CHO cells.

[0126] The anti-CD123 antibody or its antigen-binding fragment for use in the methods provided herein may be a cysteine-modified antibody or fragment. The cysteine-modified antibody can covalently bind to the cytotoxic agent of interest to form an immune complex.

[0127] As used herein, the expressions “linked to a cell binding agent” or “linked to an anti-CD123 antibody or fragment” refer to a conjugated molecule comprising at least one cytotoxic agent linked to a cell binding agent, such as an anti-CD123 antibody or fragment, via a suitable linking group or its precursor. Examples of linkers include peptide linkers.

[0128] An immune complex may contain multiple cytotoxic agents conjugated to an antibody or its antigen-binding fragment. As provided herein, in certain cases, about one to about three drug molecules, for example, cytotoxic agents, are conjugated to an anti-CD123 antibody or its antigen-binding fragment. In one embodiment, the immune complex contains one, two, or three cytotoxic agents per antibody or its antigen-binding fragment.

[0129] A composition containing an immune complex may contain immune complexes with a different number of bound cytotoxic agents per antibody or its antigen-binding fragment. Therefore, a composition containing an immune complex may contain an average number of bound cytotoxic agents per antibody or its antigen-binding fragment. In one embodiment, a pharmaceutical composition containing an anti-CD123 immune complex contains about 1 to about 3 cytotoxic agents per anti-CD123 antibody or its antigen-binding fragment, about 1.5 to about 2.5 cytotoxic agents per anti-CD123 antibody or its antigen-binding fragment, about 1.5 to about 2.1 cytotoxic agents per anti-CD123 antibody or its antigen-binding fragment, or about 1.5 to about 2.0 cytotoxic agents per anti-CD123 antibody or its antigen-binding fragment.

[0130] In certain cases, a pharmaceutical composition for use in the manner provided herein comprises, for example, an anti-CD123 immune conjugate containing about 1 to about 3 cytotoxic agents per antibody or antigen-binding fragment, the average number of cytotoxic agents per antibody or antigen-binding fragment being about 1 to about 3 (e.g., 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0).

[0131] In certain cases, the pharmaceutical composition for use in the manner provided herein comprises an anti-CD123 immune complex to which an average of about 1 ± 0.2, about 1.1 ± 0.2, about 1.2 ± 0.2, about 1.3 ± 0.2, about 1.4 ± 0.2, about 1.5 ± 0.2, about 1.6 ± 0.2, about 1.7 ± 0.2, about 1.8 ± 0.2, about 1.9 ± 0.2, about 2.0 ± 0.2, about 2.1 ± 0.2, 2.2 ± 0.2, 2.3 ± 0.2, 2.4 ± 0.2, 2.5 ± 0.2, or 2.6 ± 0.2 drug molecules (e.g., cytotoxic agents) are attached per antibody or its antigen-binding fragment. In certain embodiments, the pharmaceutical composition provided in the present invention comprises an anti-CD123 immune complex having an average of about 1.5 to 2.1 drug molecules (e.g., cytotoxic agents) per antibody.

[0132] Antibodies or antigen-binding fragments for use in this disclosure may be linked to cytotoxic agents, for example, via linkage to a Lys side-chain amino group, a Cys side-chain thiol group, or an oxidized N-terminal Ser / Thr. Examples of cytotoxic agents include DNA alkylating agents such as indolino-benzodiazepine (IGN) DNA alkylating agents. In certain cases, the cytotoxic agent is an indolino-benzodiazepine pseudodimer. In certain cases, the anti-CD123 immune complex for use in this disclosure comprises DGN549-C.

[0133] III. BCL-2 inhibitors Described herein is a method of administering an anti-CD123 immune complex, such as IMGN632, in combination with a BCL-2 inhibitor. Overexpression of BCL-2 has been demonstrated in CLL and AML cells, where BCL-2 mediates tumor cell survival and is associated with resistance to chemotherapeutic agents. BCL-2 inhibitors can reverse this effect, for example, by promoting apoptosis.

[0134] Examples of BCL-2 inhibitors include venetoclax (Venclexta®), GX15-070 (GeminX), AT-101 (Ascenta), and ABT-263 (Navitoclax, Abbott).

[0135] Venetoclax (also known as 4-(4-{[2-(4-chlorophenyl)-4,4-dimethylcyclohexy-1-en-1-yl]methyl}piperazine-1-yl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]pyridine-5-yloxy)benzamide) is a selective inhibitor of BCL-2. Venetoclax is thought to help restore the apoptotic process by directly binding to the BCL-2 protein, replacing preapoptotic proteins such as BIM, and causing mitochondrial outer membrane permeability and caspase activation. Venetoclax is the active ingredient in Venclexta®, which is available as an oral tablet.

[0136] In some embodiments, the BCL-2 inhibitor is a small molecule. In some embodiments, the BCL-2 inhibitor is venetoclax.

[0137] IV. Low-methylating agents Described herein are methods for administering an anti-CD123 immune complex, such as IMGN632, in combination with a hypomethylating agent, such as azacitidine or decitabine.

[0138] Azacitidine (also known as "4-amino-1-β-D-ribofuranosyl-s-triazine-2(1H)-one" or "5-azacitin") is a pyrimidine nucleoside analog. It is thought to induce antineoplastic activity through two mechanisms: inhibition of DNA methyltransferase at low doses, which causes hypomethylation of DNA, and direct cytotoxicity in abnormal hematopoietic cells in the bone marrow through incorporation into DNA and RNA at high doses, which leads to cell death. Azacitidine is the active ingredient in Vidaza®, and is provided in sterile form for reconstitution as a suspension for subcutaneous injection or as a solution with further dilution for intravenous administration.

[0139] Decitabine (also known as 4-amino-1-(2-deoxy-β-D-erythropentofuranosyl)-1,3,5-triazine 2(1H)-one) is an analogue of nucleoside 2'-deoxycytidine. It is thought to exert antineoplastic effects after phosphorylation, direct incorporation into DNA, and inhibition of DNA methyltransferase, leading to DNA hypomethylation and cell differentiation or apoptosis. Decitabine is the active ingredient in Dacogen®, which is supplied as a sterile lyophilized powder for reconstitution for intravenous administration.

[0140] By administering HMA in combination with an anti-CD123 immune conjugate (e.g., IMGN632), the amount and / or frequency of HMA required to achieve the same efficacy can be reduced, thereby reducing the toxicity of the therapy. Administration of HMA in combination with an anti-CD123 immune conjugate (e.g., IMGN632) can also increase the efficacy of the therapy.

[0141] In some embodiments, HMA is a low molecular weight. In some embodiments, HMA is azacitidine. In some embodiments, HMA is decitabine.

[0142] V. Pharmaceutical Compositions and Kits As provided herein, anti-CD123 immune complexes (e.g., IMGN632) can be used in combination with BCL-2 inhibitors (e.g., venetoclax) and / or hypomethylating agents (HMAs) (e.g., azacitidine or decitabine) to treat cancer.

[0143] In some embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) and a BCL-2 inhibitor (e.g., venetoclax) are contained within the same pharmaceutical composition. In some embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) and a BCL-2 inhibitor (e.g., venetoclax) are contained within two separate pharmaceutical compositions in a single kit. In other embodiments, the kit includes an anti-CD123 immune conjugate (e.g., IMGN632), as well as instructions for administering the anti-CD123 immune conjugate (e.g., IMGN632) and the BCL-2 inhibitor (e.g., venetoclax). In other embodiments, the kit includes a BCL-2 inhibitor (e.g., venetoclax), as well as instructions for administering the BCL-2 inhibitor (e.g., venetoclax) and an anti-CD123 immune conjugate (e.g., IMGN632).

[0144] In some embodiments, the anti-CD123 immune complex (e.g., IMGN632) and HMA (e.g., azacitidine or decitabine) are contained within the same pharmaceutical composition. In some embodiments, the anti-CD123 immune complex (e.g., IMGN632) and HMA (e.g., azacitidine or decitabine) are contained within two separate pharmaceutical compositions in a single kit. In other embodiments, the kit includes the anti-CD123 immune complex (e.g., IMGN632), as well as instructions for administering the anti-CD123 immune complex (e.g., IMGN632) and HMA (e.g., azacitidine or decitabine). In other embodiments, the kit includes the HMA (e.g., azacitidine or decitabine), as well as instructions for administering the HMA (e.g., azacitidine or decitabine) and the anti-CD123 immune complex (e.g., IMGN632).

[0145] In some embodiments, an anti-CD123 immune complex (e.g., IMGN632), a BCL-2 inhibitor (e.g., venetoclax), and an HMA (e.g., azacitidine or decitabine) are included in the same pharmaceutical composition. In some embodiments, the anti-CD123 immune complex (e.g., IMGN632), a BCL-2 inhibitor (e.g., venetoclax), and an HMA (e.g., azacitidine or decitabine) are included in two or three separate pharmaceutical compositions within a single kit.

[0146] In other embodiments, the kit includes an anti-CD123 immune complex (e.g., IMGN632), and instructions for administering the anti-CD123 immune complex (e.g., IMGN632) together with a BCL-2 inhibitor (e.g., venetoclax) and an HMA (e.g., azacitidine or decitabine). In other embodiments, the kit includes a BCL-2 inhibitor (e.g., venetoclax), and instructions for administering the BCL-2 inhibitor (e.g., venetoclax) together with an anti-CD123 immune complex (e.g., IMGN632) and an HMA (e.g., azacitidine or decitabine). In other embodiments, the kit includes an HMA (e.g., azacitidine or decitabine), and instructions for administering the HMA (e.g., azacitidine or decitabine) together with a BCL-2 inhibitor (e.g., venetoclax) and an anti-CD123 immune complex (e.g., IMGN632).

[0147] In other embodiments, the kit includes an anti-CD123 immune conjugate (e.g., IMGN632), a BCL-2 inhibitor (e.g., venetoclax), and instructions for administering the anti-CD123 immune conjugate (e.g., IMGN632) and the BCL-2 inhibitor (e.g., venetoclax) together with an HMA (e.g., azacitidine or decitabine). In other embodiments, the kit includes instructions for administering a BCL-2 inhibitor (e.g., venetoclax), an HMA (e.g., azacitidine or decitabine), and an anti-BCL-2 inhibitor (e.g., venetoclax), an HMA (e.g., azacitidine or decitabine) together with an anti-CD123 immune complex (e.g., IMGN632).

[0148] In certain embodiments, the pharmaceutical compositions provided herein include an anti-CD123 immune complex (e.g., IMGN632), a BCL-2 inhibitor (e.g., venetoclax), and / or an HMA (e.g., azacitidine or decitabine), as well as a pharmaceutically acceptable vehicle. These pharmaceutical compositions are used to inhibit tumor growth and treat cancer in subjects (e.g., human patients).

[0149] The pharmaceutical compositions for use provided herein may have an anti-CD123 immune conjugate (e.g., IMGN632), a BCL-2 inhibitor (e.g., venetoclax), and / or HMA (e.g., azacitidine or decitabine) of desired purity in a physiologically acceptable carrier, excipient, or stabilizer (Remington's Pharmaceutical Sciences (1990), Mack Publishing Co., Easton, PA). The acceptable carrier, excipient, or stabilizer is non-toxic to the recipient at the dosage and concentration used. (e.g., Gennaro, Remington: The Science and Practice of Pharmacy with Facts and Comparisons: Drugfacts Plus, 20th ed. (2003), Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery) Systems,7th ed.,Lippencott Williams and See Wilkins (2004), Kibbe et al., Handbook of Pharmaceutical Excipients, 3rd ed., Pharmaceutical Press (2000). Compositions used for in vivo administration can be sterilized. This can be easily achieved, for example, by filtering through a sterile filtration membrane.

[0150] In certain embodiments, a pharmaceutical composition comprising an anti-CD123 immune complex (e.g., IMGN632) and / or an HMA (e.g., azacitidine or decitabine) is formulated for parenteral (e.g., intravenous) administration.

[0151] In certain embodiments, a pharmaceutical composition containing a BCL-2 inhibitor (e.g., venetoclax) is formulated for oral administration, for example, as a tablet.

[0152] VI.How to use As provided herein, anti-CD123 immune complexes (e.g., IMGN632) may be used in combination with BCL-2 inhibitors and / or hypomethylating agents (HMAs) to treat hematological malignancies.

[0153] VI(A). Cancer Selection Cancers that can be treated by the methods provided herein include hematological malignancies. In certain embodiments, the hematological malignancy is of bone marrow origin. In certain embodiments, the hematological malignancy is of lymphoid origin. In certain embodiments, the hematological malignancy is both bone marrow and lymphoid origin. In certain embodiments, the hematological malignancy is a B-cell malignancy. In certain embodiments, the hematological malignancy is a hematological malignancy that expresses CD123.

[0154] In certain embodiments, the hematological malignancy is selected from the group consisting of acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), acute lymphoblastic leukemia (ALL), B-cell acute lymphoblastic leukemia (B-ALL), T-cell acute lymphoblastic leukemia (T-ALL), chronic myeloid leukemia in acute transformation / acute transformation phase (BP-CML), and blastic plasmacytoid dendritic cell neoplasms (BPDCN). In certain embodiments, the hematological malignancy is chronic myelomonocytic leukemia (CMML). In certain embodiments, the hematological malignancy is myelofibrosis (MF).

[0155] In certain embodiments, the hematological malignancy is a recurrent hematological malignancy. In certain embodiments, the hematological malignancy is a refractory hematological malignancy.

[0156] In certain embodiments, hematological malignancies are AML. In certain embodiments, AML is recurrent AML. In certain embodiments, AML is refractory AML. In certain embodiments, AML is not secondary AML. In certain embodiments, AML is conforming AML. In certain embodiments, AML is non-conforming AML.

[0157] In certain embodiments, the hematological malignancy is BPDCN. In certain embodiments, the BPDCN is recurrent BPDCN. In certain embodiments, the BPDCN is refractory BPDCN.

[0158] In certain embodiments, the hematological malignancy is ALL. In certain embodiments, ALL is recurrent ALL. In certain embodiments, ALL is refractory ALL.

[0159] In certain embodiments, hematological malignancies are MDS. In certain embodiments, MDS is high-risk MDS.

[0160] In a particular embodiment, the hematological malignancy is chronic myelomonocytic leukemia (CMML).

[0161] In certain embodiments, the hematological malignancy is myelofibrosis (MF).

[0162] In certain embodiments, hematological malignancies are resistant to chemotherapy.

[0163] In certain embodiments, hematological malignancies are chemotherapy-sensitive.

[0164] In certain embodiments, hematological malignancies express multidrug resistance 1 (MDR1). In certain embodiments, hematological malignancies express multidrug resistance (MDR)-associated P-glycoprotein (P-gp). In certain embodiments, hematological malignancies overexpress MDR1 and P-gp.

[0165] In certain embodiments, a subject with a hematological malignancy or a subject having a hematological malignancy has the FLT3-ITD mutation. In certain embodiments, a subject with a hematological malignancy or a subject having a hematological malignancy does not have the FLT3-ITD mutation.

[0166] In certain embodiments, hematological malignancies are present in the target population as minimal residual disease (MRD). In certain embodiments, MRD+ patients have compatible AML. In certain embodiments, MRD+ patients have incompatible AML. The methods provided herein can convert MRD+ patients to MRD- patients. The methods provided herein can also increase recurrence-free survival (e.g., median recurrence-free survival) in MRD+ patients.

[0167] In some embodiments, at least about 80% of the cells in hematological malignancies are CD123+ (determined, for example, by local flow cytometry or immunohistochemistry).

[0168] In some embodiments, it is determined prior to administration that at least 80% of the cells in the hematological malignancy are CD123-positive (e.g., by local flow cytometry or immunohistochemistry).

[0169] In certain cases, the cancer had not been treated previously.

[0170] In certain cases, the human subject has received at least one prior treatment regimen for cancer. In certain cases, the human subject has received one prior treatment regimen for cancer. In certain cases, the human subject has received two prior treatment regimens for cancer. In certain cases, the human subject has received two prior treatment regimens for cancer. In certain cases, the human subject has received six or fewer prior treatment regimens for cancer. In certain cases, the human subject has received at least one prior treatment for cancer, but no more than six prior treatment regimens.

[0171] In one case, the cancer had been previously treated with a BCL-2 inhibitor. In another case, the cancer had not been previously treated with a BCL-2 inhibitor (i.e., the patient was "BCL-2 inhibitor naive").

[0172] In one case, the cancer had been previously treated with venetoclax. In another case, the cancer had not been previously treated with venetoclax (i.e., the patient was "venetoclax naive").

[0173] In one case, the cancer had been previously treated with a hypomethylating agent. In another case, the cancer had not been previously treated with a hypomethylating agent (i.e., the patient was "hypomethylating agent naive").

[0174] In one case, the cancer had been previously treated with azacitidine. In another case, the cancer had not been previously treated with azacitidine (i.e., the patient was "azacitidine naive").

[0175] In one case, the cancer had been previously treated with decitabine. In another case, the cancer had not been previously treated with decitabine (i.e., the patient was "decitabine naive").

[0176] VI(B). Medication As provided herein, anti-CD123 immune complexes (e.g., IMGN632) may be administered in specific doses and / or at specific time intervals. Administration of anti-CD123 immune complexes (e.g., IMGN632) may be, for example, intravenous.

[0177] In certain embodiments, the anti-CD123 immune complex (e.g., IMGN632) is administered once in a 3-week (21-day) cycle. In certain embodiments, the anti-CD123 immune complex (e.g., IMGN632) is administered three times in a 3-week cycle, for example, the first dose being on day 1 (e.g., day 1), the second dose being 3 days after the first dose (e.g., day 4), and the third dose being 4 days after the second dose in the 21-day cycle (e.g., day 7). For example, when the first dose of the immune complex (e.g., IMGN632) is on day 7 of a 21-day cycle (e.g., when the immune complex is administered in combination with a BCL-2 inhibitor (e.g., venetoclax)), the immune complex (e.g., IMGN632) may be administered on days 7, 10, and 14 of the 21-day cycle.

[0178] In a particular embodiment, the anti-CD123 immune complex (e.g., IMGN632) is administered once in a 4-week (28-day) cycle. In a particular embodiment, the anti-CD123 immune complex (e.g., IMGN632) is administered three times in a 4-week cycle, for example, the first dose being on the first day (e.g., day 1), the second dose being three days after the first dose (e.g., day 4), and the third dose being four days after the second dose in the 28-day cycle (e.g., day 7). For example, when the first dose of an immune complex (e.g., IMGN632) is administered on day 7 of a 28-day cycle (e.g., when the immune complex is administered in combination with an HMA (e.g., azacitidine), or in combination with a BCL-2 inhibitor (e.g., venetoclax) and an HMA (e.g., azacitidine), the immune complex (e.g., IMGN632) may be administered on days 7, 10, and 14 of a 28-day cycle.

[0179] In certain embodiments, one cycle of treatment is therapeutically effective. In certain embodiments, two cycles of treatment are therapeutically effective. In certain embodiments, one to four cycles of treatment are therapeutically effective. In certain embodiments, two to twelve cycles of treatment are therapeutically effective.

[0180] In some embodiments, a patient can be treated in one cycle (e.g., a 21-day or 28-day cycle), for example, the immune complex is administered once or three times during the cycle. In some embodiments, a patient can be treated in at least two cycles (e.g., a 21-day or 28-day cycle), for example, the immune complex is administered once or three times per cycle. In some embodiments, a patient can be treated in at least three cycles (e.g., a 21-day or 28-day cycle), for example, the immune complex is administered once or three times per cycle. In some embodiments, a patient can be treated in at least four cycles (e.g., a 21-day or 28-day cycle), for example, the immune complex is administered once or three times per cycle. In some embodiments, a patient can be treated in at least five cycles (e.g., a 21-day or 28-day cycle), for example, the immune complex is administered once or three times per cycle. In some embodiments, a patient can be treated for at least 6 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle. In some embodiments, a patient can be treated for at least 7 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle. In some embodiments, a patient can be treated for at least 8 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle. In some embodiments, a patient can be treated for at least 9 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle. In some embodiments, a patient can be treated for at least 10 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle.In some embodiments, a patient can be treated for at least 11 cycles (e.g., 21-day or 28-day cycles), with the immune complex being administered once or three times per cycle, for example. In some embodiments, a patient can be treated for at least 12 cycles (e.g., 21-day or 28-day cycles), with the immune complex being administered once or three times per cycle, for example.

[0181] In some embodiments, a patient can be treated for 1 to 10 cycles (e.g., 21-day or 28-day cycles), for example, by administering the immune complex once or three times per cycle. In some embodiments, a patient can be treated for 2 to 10 cycles (e.g., 21-day or 28-day cycles), for example, by administering the immune complex once or three times per cycle. In some embodiments, a patient can be treated for 3 to 10 cycles (e.g., 21-day or 28-day cycles), for example, by administering the immune complex once or three times per cycle. In some embodiments, a patient can be treated for 4 to 10 cycles (e.g., 21-day or 28-day cycles), for example, by administering the immune complex once or three times per cycle. In some embodiments, a patient can be treated for 5 to 10 cycles (e.g., 21-day or 28-day cycles), for example, by administering the immune complex once or three times per cycle.

[0182] In some embodiments, patients can be treated for 1 to 12 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle. In some embodiments, patients can be treated for 2 to 12 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle. In some embodiments, patients can be treated for 3 to 12 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle. In some embodiments, patients can be treated for 4 to 12 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle. In some embodiments, patients can be treated for 5 to 12 cycles (e.g., 21-day or 28-day cycles), for example, the immune complex is administered once or three times per cycle.

[0183] In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg is administered once in a 3-week (21-day) cycle. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg is administered once in a 4-week (28-day) cycle. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg is administered once in a 3-week (21-day) cycle. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg is administered once in a 4-week (28-day) cycle.

[0184] In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 3-week (21-day) cycle at a dose of 0.015 mg / kg to approximately 0.045 mg / kg. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 3-week (21-day) cycle at a dose of approximately 0.03 mg / kg. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 4-week (28-day) cycle at a dose of 0.015 mg / kg to approximately 0.045 mg / kg. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 4-week (28-day) cycle at a dose of approximately 0.03 mg / kg.

[0185] In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of 0.045 mg / kg to approximately 0.09 mg / kg is administered once in a 3-week (21-day) cycle. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of 0.045 mg / kg to approximately 0.09 mg / kg is administered once in a 4-week (28-day) cycle.

[0186] In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 3-week (21-day) cycle at a dose of 0.045 mg / kg to approximately 0.135 mg / kg. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 4-week (28-day) cycle at a dose of 0.045 mg / kg to approximately 0.135 mg / kg.

[0187] In certain embodiments, approximately 0.015 mg / kg of anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 3-week (21-day) or 4-week (28-day) cycle. In certain embodiments, approximately 0.03 mg / kg of anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 3-week (21-day) or 4-week (28-day) cycle. In certain embodiments, approximately 0.045 mg / kg of anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 3-week (21-day) or 4-week (28-day) cycle. In certain embodiments, approximately 0.09 mg / kg of anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 3-week (21-day) or 4-week (28-day) cycle. In a particular embodiment, approximately 0.135 mg / kg of anti-CD123 immune conjugate (e.g., IMGN632) is administered once in a 3-week (21-day) or 4-week (28-day) cycle.

[0188] In certain embodiments, a total dose of 0.045 mg / kg to approximately 0.18 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) is administered over a 3-week or 4-week cycle, and this total dose is divided into three separate doses (e.g., on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14). Thus, in certain embodiments, approximately 0.015 mg / kg to approximately 0.09 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) is administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg is administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.06 mg / kg is administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.03 mg / kg is administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.03 mg / kg to approximately 0.06 mg / kg is administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14.

[0189] In certain embodiments, approximately 0.015 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) are administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14. In certain embodiments, approximately 0.03 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) are administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14. In certain embodiments, approximately 0.06 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) are administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14. In certain embodiments, approximately 0.09 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) are administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14. In certain embodiments, approximately 0.135 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) are administered three times in a 3-week or 4-week cycle, for example, on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14.

[0190] As provided herein, BCL-2 inhibitors may be administered in specific doses and / or at specific time intervals. Administration of a BCL-2 inhibitor (e.g., venetoclax) may be, for example, orally (e.g., in tablet form).

[0191] In certain embodiments, the BCL-2 inhibitor (e.g., venetoclax) is administered at a daily dose of 400 mg. In certain embodiments, the BCL-2 inhibitor (e.g., venetoclax) is administered at a daily dose of 200 mg.

[0192] In certain embodiments, a BCL-2 inhibitor (e.g., venetoclax) is administered in a 21-day cycle. The BCL-2 inhibitor (e.g., venetoclax) can be administered, for example, on days 1-7, 1-8, 1-14, 1-18, or 1-21 of a 21-day cycle. In certain embodiments, a BCL-2 inhibitor (e.g., venetoclax) is administered in a 28-day cycle. The BCL-2 inhibitor (e.g., venetoclax) can be administered, for example, on days 1-7, 1-8, 1-14, 1-18, 1-21, or 1-28 of a 28-day cycle.

[0193] In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1, 200 mg on day 2, and 400 mg on all subsequent days of the cycle, e.g., days 3-21 of a 21-day cycle or days 3-28 of a 28-day cycle.

[0194] In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1, 200 mg on day 2, and 400 mg on days 3-18 of a 21-day or 28-day cycle. In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1, 200 mg on day 2, and 400 mg on days 3-14 of a 21-day or 28-day cycle. In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1, 200 mg on day 2, and 400 mg on days 3-8 of a 21-day or 28-day cycle. In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1, 200 mg on day 2, and 400 mg on days 3-7 of a 21-day or 28-day cycle.

[0195] In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1 and 200 mg on all subsequent days of the cycle, e.g., days 2 - 21 of a 21-day cycle or days 2 - 28 of a 28-day cycle. In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1 of a 21-day cycle or a 28-day cycle and 200 mg on days 2 - 18. In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1 of a 21-day cycle or a 28-day cycle and 200 mg on days 2 - 14. In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1 of a 21-day cycle or a 28-day cycle and 200 mg on days 2 - 8. In certain embodiments, for example, in the first cycle, the BCL-2 inhibitor (e.g., venetoclax) is orally administered (PO) at 100 mg on day 1 of a 21-day cycle or a 28-day cycle and 200 mg on days 2 - 7.

[0196] In certain embodiments, the administration of the anti-CD123 immune complex (e.g., IMGN632) is initiated on day 7 after the 7th dose of the BCL-2 inhibitor (e.g., venetoclax).

[0197] The cycles of administration of the anti-CD123 immune complex and the BCL-2 inhibitor (e.g., venetoclax) (e.g., 21-day cycles or 28-day cycles) can be repeated, for example, 2 - 12 times.

[0198] In certain embodiments, the BCL-2 inhibitor (e.g., venetoclax) is administered daily. In certain embodiments, the BCL-2 inhibitor (e.g., venetoclax) is orally administered. In certain embodiments, the BCL-2 inhibitor (e.g., venetoclax) is orally administered daily.

[0199] In certain embodiments, 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily. In certain embodiments, 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally. In certain embodiments, 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily.

[0200] Following administration of anti-CD123 immune complexes and BCL-2 inhibitors (e.g., venetoclax), anti-CD123 immune complexes (e.g., IMGN632) can be administered as maintenance therapy.

[0201] As provided herein, HMAs may be administered in specific doses and / or at specific time intervals. Administration of HMAs (e.g., azacitidine or decitabine) may be, for example, subcutaneous or intravenous.

[0202] In a particular embodiment, azacitidine is 75 mg / m² 2 It can be administered subcutaneously (SC) or intravenously (IV). In certain embodiments, azacitidine is administered daily at a dose of 75 mg / m² for seven consecutive days. 2 It can be administered subcutaneously (SC) or intravenously (IV). In certain embodiments, azacitidine is administered daily at a dose of 75 mg / m² for 7 consecutive days in a 28-day cycle. 2 It can be administered subcutaneously (SC) or intravenously (IV). In certain embodiments, azacitidine is administered daily at a dose of 75 mg / m² for 5 consecutive days. 2 It can be administered subcutaneously (SC) or intravenously (IV). In certain embodiments, azacitidine is administered daily at a dose of 75 mg / m² for 5 consecutive days in a 28-day cycle. 2 It can be administered subcutaneously (SC) or intravenously (IV).

[0203] In a particular embodiment, HMA (e.g., azacitidine) is 75 mg / m² 2 It is administered in the following dose. In a particular embodiment, HMA (e.g., azacitidine) is 100 mg / m² 2 It is administered in the following dosage.

[0204] In certain embodiments, HMA (e.g., azacitidine) is administered daily for 7 days every 4 weeks. In certain embodiments, HMA (e.g., azacitidine) is administered daily for 5 days every 4 weeks.

[0205] In one particular embodiment, 75 mg / m² 2 HMA (e.g., azacitidine) is administered daily for 7 days every 4 weeks. In one particular embodiment, 100 mg / m² 2 HMA (e.g., azacitidine) is administered daily for 7 days every 4 weeks. In one particular embodiment, 75 mg / m² 2 HMA (e.g., azacitidine) is administered daily for 5 days every 4 weeks. In one particular embodiment, the dose is 100 mg / m². 2 HMA (e.g., azacitidine) is administered daily for 5 days every 4 weeks.

[0206] In one particular embodiment, 75 mg / m² 2 The HMA (e.g., azacitidine) is administered subcutaneously or intravenously daily for 7 days every 4 weeks. In certain embodiments, the dose is 100 mg / m². 2 The HMA (e.g., azacitidine) is administered subcutaneously or intravenously daily for 7 days every 4 weeks. In certain embodiments, 75 mg / m² is used. 2 The HMA (e.g., azacitidine) is administered subcutaneously or intravenously daily for 5 days every 4 weeks. In certain embodiments, the dose is 100 mg / m². 2 The HMA (e.g., azacitidine) is administered subcutaneously or intravenously daily for 5 days every 4 weeks.

[0207] After administration of anti-CD123 immune complexes and HMAs (e.g., azacitidine), anti-CD123 immune complexes (e.g., IMGN632) can be administered as maintenance therapy.

[0208] In a particular embodiment, HMA (e.g., decitabine) is 15 mg / m² 2 It is administered in the following dose. In a particular embodiment, HMA (e.g., decitabine) is 20 mg / m². 2It is administered in the following dosage.

[0209] In one particular embodiment, HMA (e.g., decitabine) is administered by intravenous infusion over 3 hours, repeated every 8 hours for 3 days, and repeated every 6 weeks. In another particular embodiment, HMA (e.g., decitabine) is administered by intravenous infusion over 1 hour for 5 days, and repeated every 4 weeks.

[0210] In one particular embodiment, 15 mg / m² 2 The HMA (e.g., decitabine) is administered by intravenous infusion over 3 hours, repeated every 8 hours for 3 days, and every 6 weeks. In a particular embodiment, 20 mg / m² 2 The HMA (e.g., decitabine) is administered by intravenous infusion over one hour for five days, repeated every four weeks.

[0211] After administration of anti-CD123 immune complexes and HMAs (e.g., decitabine), anti-CD123 immune complexes (e.g., IMGN632) can be administered as maintenance therapy.

[0212] In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of 0.03 mg / kg is administered once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered intravenously once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily. In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of 0.03 mg / kg is administered intravenously once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily. The BCL-2 inhibitor (e.g., venetoclax) may be administered daily on days 1-7, 1-8, 1-14, 1-18, or 1-21 of a 21-day (3-week) cycle. BCL-2 inhibitors (e.g., venetoclax) may be administered daily on days 1–7, 1–8, 1–14, 1–18, 1–21, or 1–28 of a 28-day (4-week) cycle.

[0213] In certain embodiments, 0.015 mg / kg to approximately 0.135 mg / kg (e.g., 0.135 mg / kg) of anti-CD123 immune conjugates (e.g., IMGN632) is administered once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily. In certain embodiments, 0.03 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) is administered once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily. In certain embodiments, 0.015 mg / kg to approximately 0.135 mg / kg (e.g., 0.135 mg / kg) of anti-CD123 immune conjugates (e.g., IMGN632) is administered intravenously once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily. In certain embodiments, 0.03 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) are administered intravenously once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily. In certain embodiments, 0.03 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) are administered intravenously once every three or four weeks, and 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily. The BCL-2 inhibitor (e.g., venetoclax) can be administered daily on days 1-7, 1-8, 1-14, 1-18, or 1-21 of a 21-day (3-week) cycle. BCL-2 inhibitors (e.g., venetoclax) can be administered daily on days 1-7, 1-8, 1-14, 1-18, 1-21, or 1-28 of a 28-day (4-week) cycle.

[0214] In a particular embodiment, a total dose of 0.045 mg / kg to about 0.18 mg / kg of anti-CD123 immune conjugates (e.g., IMGN632) is administered over a 3-week cycle, with this total dose divided into three separate doses (e.g., on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14), and a BCL-2 inhibitor (e.g., venetoclax) is administered daily. Thus, in a particular embodiment, about 0.015 mg / kg to about 0.06 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) of anti-CD123 immune conjugates (e.g., IMGN632) is administered three times in a 21-day cycle (e.g., on days 7, 10, and 14 of the 21-day cycle), and a BCL-2 inhibitor (e.g., venetoclax) is administered daily. In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) is administered intravenously in doses of approximately 0.015 mg / kg to approximately 0.06 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) three times in a 21-day cycle (e.g., on days 7, 10, and 14 of the 21-day cycle), and a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily. The BCL-2 inhibitor (e.g., venetoclax) can be administered in doses of 100 mg, 200 mg, and / or 400 mg. The BCL-2 inhibitor (e.g., venetoclax) can be administered in doses of 100 mg on day 1, 200 mg on day 2, and 400 mg on days 3 through 21 of the 21-day cycle (e.g., the first 21-day cycle). BCL-2 inhibitors (e.g., venetoclax) can be administered in doses of 100 mg on day 1 of a 21-day cycle (e.g., the first 21-day cycle) and 200 mg on days 2 through 21. BCL-2 inhibitors (e.g., venetoclax) can be administered as an oral daily dose of 400 mg on all days of a 21-day cycle (e.g., days 1 through 21). BCL-2 inhibitors (e.g., venetoclax) can be administered as an oral daily dose of 200 mg on all days of a 21-day cycle (e.g., days 1 through 21).

[0215] In one particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) at a dose of 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered once every four weeks, followed by 75 mg / m². 2 or 100 mg / m² 2 The HMA (e.g., azacitidine) is administered daily for 7 days every 4 weeks. In a particular embodiment, 0.03 mg / kg of anti-CD123 immune complex (e.g., IMGN632) is administered once every 4 weeks, and 75 mg / m² is administered. 2 or 100 mg / m² 2 HMA (e.g., azacitidine) is administered daily for 7 days every 4 weeks. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at a dose of 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered intravenously once every 4 weeks, and 75 mg / m² is administered. 2 or 100 mg / m² 2 The HMA (e.g., azacitidine) is administered subcutaneously or intravenously daily for 7 days every 4 weeks. In certain embodiments, 0.03 mg / kg of anti-CD123 immune conjugate (e.g., IMGN632) is administered intravenously once every 4 weeks, totaling 75 mg / m². 2 or 100 mg / m² 2 Administer HMA (e.g., azacitidine) subcutaneously or intravenously daily for 7 days every 4 weeks.

[0216] In one particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) at a dose of 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered once every four weeks, followed by 75 mg / m². 2 or 100 mg / m² 2 The HMA (e.g., azacitidine) is administered daily for 5 days every 4 weeks. In a particular embodiment, 0.03 mg / kg of anti-CD123 immune complex (e.g., IMGN632) is administered once every 4 weeks, and 75 mg / m² is administered. 2 or 100 mg / m² 2Administer the HMA (e.g., azacitidine) daily for 5 days every 4 weeks. In certain embodiments, administer an anti-CD123 immune complex (e.g., IMGN632) at 0.015 mg / kg to about 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) intravenously once every 4 weeks, 75 mg / m 2 or 100 mg / m 2 Administer the HMA (e.g., azacitidine) subcutaneously or intravenously daily for 5 days every 4 weeks. In certain embodiments, administer an anti-CD123 immune complex (e.g., IMGN632) at 0.03 mg / kg intravenously once every 4 weeks, 75 mg / m 2 or 100 mg / m 2 Administer the HMA (e.g., azacitidine) subcutaneously or intravenously daily for 5 days every 4 weeks.

[0217] In certain embodiments, administer an anti-CD123 immune complex (e.g., IMGN632) at 0.015 mg / kg to about 0.135 mg / kg (e.g., 0.135 mg / kg) once every 4 weeks, 75 mg / m 2 or 100 mg / m 2 Administer the HMA (e.g., azacitidine) daily for 7 days every 4 weeks. In certain embodiments, administer an anti-CD123 immune complex (e.g., IMGN632) at 0.015 mg / kg to about 0.135 mg / kg (e.g., 0.135 mg / kg) intravenously once every 4 weeks, 75 mg / m 2 or 100 mg / m 2 Administer the HMA (e.g., azacitidine) subcutaneously or intravenously daily for 7 days every 4 weeks.

[0218] In certain embodiments, administer an anti-CD123 immune complex (e.g., IMGN632) at 0.015 mg / kg to about 0.135 mg / kg (e.g., 0.135 mg / kg) once every 4 weeks, 75 mg / m 2 or 100 mg / m 2HMA (e.g., azacitidine) is administered daily for 5 days every 4 weeks. In certain embodiments, 0.015 mg / kg to approximately 0.135 mg / kg (e.g., 0.135 mg / kg) of anti-CD123 immune complex (e.g., IMGN632) is administered intravenously once every 4 weeks, and 75 mg / m² is administered. 2 or 100 mg / m² 2 Administer HMA (e.g., azacitidine) subcutaneously or intravenously for 5 days every 4 weeks.

[0219] In a particular embodiment, a total dose of 0.045 mg / kg to approximately 0.18 mg / kg of anti-CD123 immune complex (e.g., IMGN632) is administered over a 4-week cycle, and this total dose is divided into three separate doses (e.g., on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14), totaling 75 mg / m². 2 or 100 mg / m² 2 HMA (e.g., azacitidine) is administered daily for 7 days every 4 weeks. In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.06 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) is administered three times in a 21-day cycle (e.g., on days 7, 10, and 14 of a 28-day cycle), and 75 mg / m² is administered. 2 or 100 mg / m² 2 HMA (e.g., azacitidine) is administered daily for 7 days every 4 weeks. In certain embodiments, anti-CD123 immune complexes (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.06 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) are administered intravenously three times in a 28-day cycle (e.g., on days 7, 10, and 14 of the 28-day cycle), totaling 75 mg / m². 2 or 100 mg / m² 2 Administer HMA (e.g., azacitidine) subcutaneously or intravenously daily for 7 days every 4 weeks.

[0220] In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered once every three weeks, followed by 15 mg / m². 2 The HMA (e.g., decitabine) is administered over 3 hours, repeated every 8 hours for 3 days, and repeated every 6 weeks. In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) of approximately 0.03 mg / kg is administered once every 3 weeks, and 15 mg / m² is administered. 2 The HMA (e.g., decitabine) is administered over 3 hours, repeated every 8 hours for 3 days, and repeated every 6 weeks. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered intravenously once every 3 weeks, and 15 mg / m² is administered. 2 The HMA (e.g., decitabine) is administered by intravenous infusion over 3 hours, repeated every 8 hours for 3 days, and repeated every 6 weeks. In a particular embodiment, approximately 0.03 mg / kg of anti-CD123 immune complex (e.g., IMGN632) is administered intravenously once every 3 weeks, and 15 mg / m² is administered. 2 The HMA (e.g., decitabine) is administered intravenously over 3 hours, repeated every 8 hours for 3 days, and then every 6 weeks.

[0221] In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) of approximately 0.015 mg / kg to approximately 0.135 mg / kg (e.g., 0.135 mg / kg) is administered once every three weeks, followed by 15 mg / m². 2 The HMA (e.g., decitabine) is administered over 3 hours, repeated every 8 hours for 3 days, and repeated every 6 weeks. In a particular embodiment, 0.015 mg / kg to about 0.135 mg / kg (e.g., 0.135 mg / kg) of anti-CD123 immune complex (e.g., IMGN632) is administered intravenously once every 3 weeks, and 15 mg / m² is administered. 2The HMA (e.g., decitabine) is administered intravenously over 3 hours, repeated every 8 hours for 3 days, and then every 6 weeks.

[0222] In a particular embodiment, a total dose of 0.045 mg / kg to approximately 0.18 mg / kg of anti-CD123 immune complex (e.g., IMGN632) is administered over a 4-week cycle, and this total dose is divided into three separate doses (e.g., on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14), with 15 mg / m² being administered. 2 HMA (e.g., decitabine) is administered over a 3-hour period, repeated every 8 hours for 3 days, and repeated every 6 weeks. In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) is administered three times in a 21-day cycle (e.g., on days 7, 10, and 14 of the 21-day cycle), and 15 mg / m² is administered. 2 The HMA (e.g., decitabine) is administered over 3 hours, repeated every 8 hours for 3 days, and repeated every 6 weeks. In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.135 mg / kg (e.g., 0.135 mg / kg) is administered three times in a 21-day cycle (e.g., on days 7, 10, and 14 of the 21-day cycle), and 15 mg / m² is administered. 2 The HMA (e.g., decitabine) is administered over 3 hours, repeated every 8 hours for 3 days, and repeated every 6 weeks. In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.06 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered three times in a 21-day cycle (e.g., on days 7, 10, and 14 of the 21-day cycle), with a dose of 15 mg / m². 2 The HMA (e.g., decitabine) is administered intravenously over 3 hours, repeated every 8 hours for 3 days, and then every 6 weeks.

[0223] In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at about 0.015 mg / kg to about 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered once every four weeks, and 20 mg / m 2 of HMA (e.g., decitabine) is administered over 1 hour for 5 days and repeated every four weeks. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at about 0.03 mg / kg is administered once every four weeks, and 20 mg / m 2 of HMA (e.g., decitabine) is administered over 1 hour for 5 days and repeated every four weeks. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at about 0.015 mg / kg to about 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered intravenously once every four weeks, and 20 mg / m 2 of HMA (e.g., decitabine) is administered by intravenous infusion over 1 hour for 5 days and repeated every four weeks. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at about 0.03 mg / kg is administered intravenously once every four weeks, and 20 mg / m 2 of HMA (e.g., decitabine) is administered by intravenous infusion over 1 hour for 5 days and repeated every four weeks.

[0224] In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at about 0.015 mg / kg to about 0.135 mg / kg (e.g., 0.135 mg / kg) is administered once every four weeks, and 20 mg / m 2 of HMA (e.g., decitabine) is administered over 1 hour for 5 days and repeated every four weeks. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at about 0.015 mg / kg to about 0.135 mg / kg (e.g., 0.135 mg / kg) is administered intravenously once every four weeks, and 20 mg / m 2 of HMA (e.g., decitabine) is administered by intravenous infusion over 1 hour for 5 days and repeated every four weeks.

[0225] In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at a total dose of 0.045 mg / kg to about 0.18 mg / kg is administered over the course of a 4-week cycle, and this total dose is divided into three separate administrations (e.g., on days 1, 4, and 8 of the cycle, or days 7, 10, and 14), 20 mg / m 2 of HMA (e.g., decitabine) is administered for 5 days over 1 hour and repeated every 4 weeks. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at about 0.015 mg / kg to about 0.06 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) is administered three times in a 28-day cycle (e.g., on days 7, 10, and 14 of a 21-day cycle), 20 mg / m 2 of HMA (e.g., decitabine) is administered for 5 days over 1 hour and repeated every 4 weeks. In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at about 0.015 mg / kg to about 0.06 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) is intravenously administered three times in a 28-day cycle (e.g., on days 7, 10, and 14 of a 21-day cycle), 20 mg / m 2 of HMA (e.g., decitabine) is administered by intravenous infusion for 5 days over 1 hour and repeated every 4 weeks.

[0226] In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at 0.015 mg / kg to about 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered once every 4 weeks, and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily (e.g., 100 mg on day 1, 200 mg on day 2, and 400 mg on days 3 - 28 of cycle 1, and 400 mg daily on days 1 - 28 in subsequent cycles), 75 mg / m 2 or 100 mg / m 2HMA (e.g., azacitidine) is administered daily (e.g., on days 1-7). In a particular embodiment, 0.03 mg / kg of anti-CD123 immune conjugate (e.g., IMGN632) is administered once every four weeks, and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-28, and 400 mg daily on days 1-28 of subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2 HMA (e.g., azacitidine) is administered daily (e.g., on days 1-7). In certain embodiments, an anti-CD123 immune conjugate (e.g., IMGN632) at a dose of 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.045, or 0.09 mg / kg) is administered intravenously once every four weeks, and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-28, and 400 mg daily on days 1-28 of subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2 HMA (e.g., azacitidine) is administered subcutaneously or intravenously daily for 7 days (e.g., on days 1-7). In certain embodiments, 0.03 mg / kg of anti-CD123 immune conjugate (e.g., IMGN632) is administered intravenously once every 4 weeks, and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-28, and 400 mg daily on days 1-28 of subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2 Administer HMA (e.g., azacitidine) subcutaneously or intravenously daily for 7 days (e.g., on days 1-7).

[0227] In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) at a dose of 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.03, 0.045, or 0.09 mg / kg) is administered once every four weeks, and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-7, 3-8, 3-14, 3-18, or 3-21, or 3-28, and 400 mg daily on days 1-7, 1-8, 1-14, 1-18, 1-21, or 1-28 in subsequent cycles), along with 75 mg / m². 2 or 100 mg / m² 2 Administer HMA (e.g., azacitidine) daily (e.g., on days 1-5 or 1-7). In a particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) is administered intravenously once every four weeks at a dose of 0.015 mg / kg to approximately 0.09 mg / kg (e.g., 0.015, 0.03, 0.045, or 0.09 mg / kg), and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-7, 3-8, 3-14, 3-18, or 3-21, or 3-28, and 400 mg daily on days 1-7, 1-8, 1-14, 1-18, 1-21, or 1-28 in subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2 Administer HMA (e.g., azacitidine) subcutaneously or intravenously daily for 7 days (e.g., on days 1-5 or 1-7).

[0228] In one particular embodiment, an anti-CD123 immune complex (e.g., IMGN632) is administered once every four weeks at a dose of 0.015 mg / kg to approximately 0.135 mg / kg (e.g., 0.135 mg / kg), and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-28, and 400 mg daily on days 1-28 of subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2 HMA (e.g., azacitidine) is administered daily (e.g., on days 1-7). In certain embodiments, 0.015 mg / kg to about 0.135 mg / kg (e.g., 0.135 mg / kg) of anti-CD123 immune complex (e.g., IMGN632) is administered intravenously once every four weeks, and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered orally daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-28, and 400 mg daily on days 1-28 of subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2 Administer HMA (e.g., azacitidine) subcutaneously or intravenously daily for 7 days (e.g., on days 1-7).

[0229] In a particular embodiment, a total dose of 0.045 mg / kg to approximately 0.18 mg / kg of anti-CD123 immune complexes (e.g., IMGN632) is administered over a 4-week cycle, the total dose of which is divided into three separate doses (e.g., on days 1, 4, and 8 of the cycle, or on days 7, 10, and 14), and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-28, and 400 mg on days 1-28 of subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2HMA (e.g., azacitidine) is administered daily for 7 days (e.g., on days 1-7). In certain embodiments, an anti-CD123 immune complex (e.g., IMGN632) at approximately 0.015 mg / kg to approximately 0.06 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) is administered three times in a 28-day cycle (e.g., on days 7, 10, and 14 of the 28-day cycle), and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) is administered daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-28, and 400 mg daily on days 1-28 of subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2 HMA (e.g., azacitidine) is administered daily for 7 days (e.g., on days 1-7). In certain embodiments, anti-CD123 immune complexes (e.g., IMGN632) at a dose of approximately 0.015 mg / kg to approximately 0.06 mg / kg (e.g., 0.015, 0.03, or 0.06 mg / kg) are administered intravenously three times in a 28-day cycle (e.g., on days 7, 10, and 14 of the 28-day cycle), and 100, 200, and / or 400 mg of a BCL-2 inhibitor (e.g., venetoclax) are administered orally daily (e.g., 100 mg on day 1 of cycle 1, 200 mg on day 2, and 400 mg on days 3-28, and 400 mg daily on days 1-28 of subsequent cycles), and 75 mg / m² 2 or 100 mg / m² 2 Administer HMA (e.g., azacitidine) subcutaneously or intravenously daily for 7 days (e.g., on days 1-7).

[0230] Following administration of anti-CD123 immune complexes, BCL-2 inhibitors, and HMAs (e.g., azacitidine or decitabine), anti-CD123 immune complexes (e.g., IMGN632) can be administered as maintenance therapy.

[0231] In one case, the immune complex that binds to CD123 (e.g., IMGN632) and the BCL-2 inhibitor are administered simultaneously. In another case, the anti-CD123 immune complex (e.g., IMGN632) and the BCL-2 inhibitor are administered in separate pharmaceutical compositions. In yet another case, the anti-CD123 immune complex (e.g., IMGN632) and the BCL-2 inhibitor are administered sequentially. In yet another case, the BCL-2 inhibitor is administered over a period of time, after which both the BCL-2 inhibitor and the anti-CD123 immune complex (e.g., IMGN632) are administered (either simultaneously or sequentially). In such cases, the HMA may, at its discretion, be administered simultaneously (in the same or separate pharmaceutical compositions) or sequentially with the anti-CD123 immune complex (e.g., IMGN632) or the BCL-2 inhibitor.

[0232] In one case, the immune complex that binds to CD123 (e.g., IMGN632) and HMA are administered simultaneously. In another case, the anti-CD123 immune complex (e.g., IMGN632) and HMA are administered in the same pharmaceutical composition. In another case, the anti-CD123 immune complex (e.g., IMGN632) and HMA are administered in separate pharmaceutical compositions. In another case, the anti-CD123 immune complex (e.g., IMGN632) and HMA are administered sequentially. In yet another case, HMA is administered over a period of time, after which both HMA and the anti-CD123 immune complex (e.g., IMGN632) are administered (either simultaneously or sequentially). In such cases, the BCL-2 inhibitor may, optionally, be administered simultaneously or sequentially with the anti-CD123 immune complex (e.g., IMGN632) or HMA.

[0233] VI(C). Evaluation and Monitoring In certain embodiments, a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA is useful for inhibiting tumor growth. In certain embodiments, a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA is useful for reducing tumor volume. In certain embodiments, a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA is useful for increasing survival rates.

[0234] For example, in some embodiments, treatment with a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA results in %T / C values ​​of less than approximately 50%, less than approximately 45%, less than approximately 40%, less than approximately 35%, less than approximately 30%, less than approximately 25%, less than approximately 20%, less than approximately 15%, less than approximately 10%, or less than approximately 5%.

[0235] In certain embodiments, a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA can inhibit tumor growth in an EOL-1 subcutaneous xenograft model. In certain embodiments, a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA can inhibit tumor growth in a KG-1 subcutaneous xenograft model.

[0236] In certain embodiments, a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA can increase survival rates in a MOLM-1 disseminated xenograft model. In certain embodiments, a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA can increase survival rates in an MV4-11 disseminated xenograft model.

[0237] In certain embodiments, a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA produces a synergistic effect.

[0238] In certain embodiments, administration of a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA does not produce more toxicity than administration of a BCL-2 inhibitor and / or HMA alone. In some embodiments, administration of a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA does not produce more toxicity than administration of an anti-CD123 immune complex. In some embodiments, administration of a combination of an anti-CD123 immune complex (e.g., IMGN632) and a BCL-2 inhibitor and / or HMA does not produce more toxicity than administration of either an anti-CD123 immune complex or a BCL-2 inhibitor and / or HMA alone.

[0239] VI(D).Additional therapy In some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) disclosed herein have received prior treatment with corticosteroids. Therefore, in some embodiments, the methods provided herein include administering corticosteroids to the patient before administering anti-CD123 immune complexes to the patient. In certain cases, the corticosteroid may be selected from the group consisting of prednisone, prednisolone, methylprednisolone, beclametasone, betamethasone, dexamethasone, fludrocortisone, hydrocortisone, and triamcinolone. In certain cases, the corticosteroid is administered intravenously. In certain cases, the steroid is administered orally.

[0240] For example, in some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) as disclosed herein received prior treatment with diphenhydramine. In some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) as disclosed herein received prior treatment with 25-50 mg of diphenhydramine. In some embodiments, diphenhydramine is administered intravenously. In some embodiments, diphenhydramine is administered orally. Therefore, in some embodiments, the method provided herein includes administering diphenhydramine to the patient before administering the anti-CD123 immune complex to the patient.

[0241] In some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) as disclosed herein received prior treatment with acetaminophen. In some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) as disclosed herein received prior treatment with 325 to 650 mg of acetaminophen. In some embodiments, acetaminophen is administered intravenously. In some embodiments, acetaminophen is administered orally. Therefore, in some embodiments, the method provided in the present invention includes administering acetaminophen to the patient before administering the anti-CD123 immune complex to the patient.

[0242] In some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) as disclosed herein received prior treatment with paracetamol. In some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) as disclosed herein received prior treatment with 325 to 650 mg of paracetamol. In some embodiments, paracetamol is administered intravenously. In some embodiments, paracetamol is administered orally. Therefore, in some embodiments, the method provided in the present invention includes administering paracetamol to the patient before administering the anti-CD123 immune complex to the patient.

[0243] In some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) as disclosed herein received prior treatment with dexamethasone. In some embodiments, patients receiving anti-CD123 immune complexes in combination with a BCL-2 inhibitor (e.g., venetoclax) and / or a hypomethylating agent (HMA) (e.g., azacitidine or decitabine) as disclosed herein received prior treatment with 8 mg of dexamethasone. In some embodiments, dexamethasone is administered intravenously. In some embodiments, dexamethasone is administered orally. Therefore, in some embodiments, the method provided in the present invention includes administering dexamethasone to the patient before administering the anti-CD123 immune complex to the patient.

[0244] Embodiments of the present disclosure can be further defined by reference to the following non-limiting embodiments. It will be apparent to those skilled in the art that many modifications to both materials and methods can be made without departing from the scope of the present disclosure. [Examples]

[0245] The examples and embodiments described herein are provided for illustrative purposes only, and it will be understood that various modifications or changes in light of them are suggested to those skilled in the art and are included within the spirit and scope of this application.

[0246] Example 1 In vitro trial of IMGN632 and venetoclax combination in AML cell lines The activity of IMGN632 alone, venetoclax alone, and the combination of IMGN632 and venetoclax was investigated in in vitro cytotoxicity assays in four different acute myeloid leukemia (AML) cell lines: EOL-1, KG-1, Molm-13, and MV4-11.

[0247] EOL-1, KG-1, Molm-13, and MV4-11 cell lines were obtained via a low-frequency pathway (less than 10 pathways), and cell cultures were maintained according to the recommendations of their suppliers. Cells were collected from cell cultures in the logarithmic growth phase, counted by an automated hemocytometer, and uniformly distributed into wells in a 96-well plate so that each well contained 5,000 viable cells. Each well was encapsulated with an Fc receptor blockade, non-mammalian targeted chKTI monoclonal antibody so that the final well volume (200 μL) contained chKTI at 100 nM. Dose ranges for IMGN632 alone, venetoclax alone, and IMGN632 + venetoclax (with their respective DMSO controls in the case of venetoclax) were then prepared in media appropriate to the cell lines being assayed.

[0248] The diluent was prepared and added to the cells in the 96-well plates on the same day the cells were seeded. The cells and drugs were added such that the final well volume was always 200 μL. Each treatment condition was prepared in three separate batches for each assay run. The treated 96-well plates were left in a dark 37 °C incubator with 90% humidity and 10% CO2 for 4 days (EOL-1, Molm-13; MV4-11) or 5 days (KG-1). On day 4 or 5, 20 μL of WST-8 was added to each well and the 96-well plates were left in the incubator for 2 hours (EOL-1; Molm-13) to 7 hours (MV4-11; KG-1) to develop. At development, the plates were read on a spectrophotometer and the readings were taken at an absorbance of 650 nm. Each dose-setting assay was run once. Each combination assay was repeated three times (EOL-1, Molm-13, MV4-11) or four times (KG-1).

[0249] Two-stage calculations were performed on the collected data. The mean control absorbance of the medium only was subtracted from each condition containing cells, and the background-corrected absorbance was indexed to the mean background-corrected absorbance of their untreated control counterparts. These calculated values were plotted in GraphPad Prism as the mean and standard deviation of technical replicates across the execution of the biological replicate assays. The same final calculated values entered into GraphPad were further processed for the evaluation of synergistic effects via CalcuSyn. The mean “survival rate” from each condition was subtracted from 1 to obtain the mean “inhibition rate” (Fa) from each condition. These Fa values were entered into CalcuSyn along with the drug doses (NB: only values of 0 < x < 1 are usable “x”). Using the input values, both the combination index (CI), which measures the observed drug and the expected effect, and the normalized inhibition rate values used in the combination isobologram were calculated.

[0250] [[ID=?]]<[ The results of in vitro cytotoxicity assays of IMGN632 and venetoclax alone are shown in Figures 2A-2D. A broad dose range of IMGN632 (Figure 2A) and venetoclax (Figure 2B) was tested for efficacy against three AML cell lines (EOL-1, MV4-11, and KG-1). Doses near the inflection points of these sensitivity curves were selected for use in the IMGN632 and venetoclax combination assay. A narrower dose range of IMGN632 (Figure 2C) and venetoclax (Figure 2D) was performed as a mono-control for the combination assay, and these doses represent the dynamic range of efficacy. Figures 3A-3D show the results of the IMGN632 and venetoclax combination evaluated in four AML cell lines: EOL-1 (Figure 3A), MV4-11 (Figure 3B), KG-1 (Figure 3C), and Molm-13 (Figure 3D). In each of the four AML cell lines evaluated, IMGN632 and venetoclax killed more cells in combination than IMGN632 alone, in a dose-dependent manner. Figures 4A–4L show the readouts of the combined efficacy for the IMGN632 and venetoclax combination, comparing the observed combined results in Figures 3A–3D with the combined results expected based on the monotherapy results in Figures 2C and 2D. For each IMGN632 and venetoclax dose pairing, a combined index value and a normalized relative effect value were generated. Results were plotted on a normalized scale (Figures 4C, 4F, 4I, and 4L) to show combined efficacy as a function of drug dose (Figures 4A, 4D, 4G, and 4J), combined efficacy as a function of therapeutic effect (Figures 4B, 4E, 4H, and 4K), and combined efficacy against expected dose effect. Each row represents results from a single cell line: EOL-1 (Figures 4A-4C), MV4-11 (Figures 4D-4F), KG-1 (Figures 4G-4I), or Molm-13 (Figures 4J-4L).

[0251] Results from in vitro cytotoxicity assays of AML cell lines treated with IMGN632 and venetoclax demonstrate the additive versus synergistic combined cytotoxic effect of this combination.

[0252] Example 2 Materials and methods used in subcutaneous and disseminated xenograft models For all subcutaneous xenograft models, mouse body weight was measured twice a week, and clinical signs were monitored throughout the study period. Animals were euthanized when hind leg paralysis was present, body weight decreased by more than 20% from pre-treatment weight, tumor ulcers developed, or any signs of distress were observed.

[0253] The tumor volume of the subcutaneous xenograft model was measured in three dimensions using calipers 1-2 times per week. Tumor volume was expressed in mm using the formula V = length × width × height × 1 / 2. 3 This was expressed as (Tomayko and Reynolds, Cancer Chemother. Pharmacol. 24:148-54 (1989)). Activity was evaluated as described in Bissery et al. Cancer Res. 51:4845-52 (1991).

[0254] Tumor growth inhibition (T / C ratio) was evaluated in a subcutaneous xenograft model using the following formula: T / C (%) = (median treated tumor volume / median control tumor volume) × 100%. Tumor volumes were simultaneously determined for the treatment (T) group and vehicle control (C) group when the vehicle control tumor volume reached a predetermined size (Bissery et al. Cancer, Res. 51:4845-52 (1991). Tumor-free mice (0 mm) 3 The median daily tumor volume was determined for each treatment group, including the following factors. According to the National Cancer Institute (NCI) criteria, T / C ≤ 42% represents the minimum level of antitumor activity. T / C < 10% is considered a high level of antitumor activity.

[0255] For all seeded xenograft models, mouse body weight was measured twice a week, and clinical signs were monitored throughout the study period. The measured endpoint was survival rate. Animals were euthanized when hind leg paralysis was present, body weight decreased by more than 20% from pre-treatment weight, a visible tumor appeared, or any signs of distress were observed. Spontaneous deaths were recorded when they were observed.

[0256] In the disseminated xenograft model, tumor growth retardation was calculated as TC, where T is the median survival time (days) in the treatment group and C is the median survival time (days) in the vehicle control group. The percentage increased lifespan (%ILS) of the disseminated model was calculated using the following formula: %ILS = (TC) / C × 100%. Antitumor activity was evaluated according to the NCI criteria for the disseminated model: ILS < 25% is inactive, ILS ≥ 25% is minimal activity, ILS > 40% is active, and ILS ≥ 50% is highly active.

[0257] Example 3 In vivo efficacy of the combination of IMGN632 (single dose) and venetoclax (QD x 28) in an EOL-1 subcutaneous model. To test the efficacy of IMGN632, venetoclax, or a combination of these two drugs in vivo for their ability to reduce tumor burden, subcutaneous tumor models were used as described in the protocol below.

[0258] Each female athymic nude mouse was given 1 × 10⁶ mice in 100 μl of 50% Matrigel:50% serum-free medium (v / v). 7 One EOL-1 cell (human AML cell line) was subcutaneously inoculated into the right flank. On day 9, 24 hours before administration of the complex therapy (either alone or in combination with venetoclax), all mice in these treatment groups were intraperitoneally injected with 150 mg / kg of non-targeted chKTI antibody to block Fc receptors on EOL-1 AML cells and prevent nonspecific uptake of the complex. On day 9 after EOL-1 inoculation, the mice were randomized to test groups based on tumor volume.

[0259] Ten days after EOL-1 inoculation, mice received a single intravenous injection into the lateral tail vein of either the vehicle or 1 μg / kg (by DGN549; 0.080 mg / kg by huCD123Ab) IMGN632, or 0.5 μg / kg (by DGN549; 0.040 mg / kg by huCD123Ab) IMGN632. Venetoclax administration was also started on day 10, and mice receiving venetoclax received a single oral dose of 100 mg / kg of venetoclax on each day from days 10 to 37, for a total of 28 consecutive days, including after cell transplantation. In the combination group, mice received both IMGN632 and venetoclax as outlined above.

[0260] The results are reported in Table 4 (below) and Figure 5.

[0261] A single dose of IMGN632 at 1 μg / kg or 0.5 μg / kg resulted in T / C values ​​of 10.7% (active) or 45.0% (inactive), respectively, and at the end of the study (day 85), 2 / 6 or 0 / 6 long-term complete regression (tumor-free survivors, TFS), respectively.

[0262] A regimen of venetoclax 100 mg / kg once daily (QD) for 28 days (×28) was inactive, with a total cerebrospinal fluid ratio (T / C) of 69.1% and a total cerebrospinal fluid ratio (TFS) of 1 / 6 on day 85.

[0263] The combination of a single dose of 1 μg / kg IMGN632 with a venetoclax QD×28 regimen was highly active, resulting in a T / C ratio of 0% and a TFS of 4 / 6 at day 85, which was 2 TFS higher than that achieved with IMGN632 monotherapy, demonstrating the additional benefit of combination therapy. The combination of a single dose of 0.5 μg / kg IMGN632 with a venetoclax QD×28 regimen was also active, resulting in a T / C ratio of 21.1% and a TFS of 1 / 6 at day 85, which was a lower %T / C value than that produced by either 0.5 ug / kg IMGN632 monotherapy (45%) or venetoclax monotherapy (69.1%), indicating that this combination was able to slow tumor growth more effectively than either of the corresponding monotherapies. [Table 4]

[0264] Example 4 In vivo efficacy of the combination of IMGN632 (QW x 3) and venetoclax (QD x 28) in a KG-1 subcutaneous model. To test the efficacy of IMGN632, venetoclax, or a combination of these two drugs in vivo for their ability to reduce tumor burden, a subcutaneous tumor model was used as described in the protocol below.

[0265] Each female C.B17 SCID mouse was given 1 × 10⁶ units in 100 μl of 50% Matrigel:50% serum-free medium (v / v). 7 One KG-1 cell line (a relatively IMGN632-resistant human AML cell line) was subcutaneously inoculated into the right flank. On days 17, 24, and 31, 24 hours prior to each administration day of the conjugate therapy (either alone or in combination with venetoclax), all mice in these therapy groups were intraperitoneally injected with 150 mg / kg of non-targeted chKTI antibody to block Fc receptors on KG-1 AML cells and prevent nonspecific uptake of the conjugate. On day 17 after KG-1 inoculation, mice were randomized to test groups based on tumor volume.

[0266] On days 18, 25, and 32 after KG-1 inoculation, mice received a single intravenous injection into the lateral tail vein of either vehicle, 3 μg / kg (by DGN549; 0.24 mg / kg by huCD123Ab) IMGN632, or 10 μg / kg (by DGN549; 0.80 mg / kg by huCD123Ab) IMGN632. Venetoclax administration was also started on day 18, and mice receiving venetoclax received a single oral dose of 100 mg / kg of venetoclax daily for 28 consecutive days (days 18-45). In the combination group, mice received both IMGN632 and venetoclax, as outlined above.

[0267] The results are reported in Table 5 (below) and Figure 6.

[0268] In this study, the QW×3 regimen with IMGN632 at 3 μg / kg was inactive, with a T / C ratio of 78% and a long-term complete regression (tumor-free survival, TFS) of 0 / 6 at the end of the study (day 56). Similarly, the QW×3 regimen with IMGN632 at 10 μg / kg was inactive in this study, with a T / C ratio of 73% and a TFS of 0 / 6. The venetoclax 100 mg / kg once daily (QD) regimen for 28 days (×28) was also inactive, with a T / C ratio of 82% and a TFS of 0 / 6.

[0269] The combination of 3 μg / kg IMGN632, QW x 3, and venetoclax QD x 28 regimens was also inactive, with a T / C ratio of 76% and a TFS of 0 / 6. However, the combination of 10 μg / kg IMGN632, QW x 3, and venetoclax QD x 28 regimens was also active, with a T / C ratio of 40% and a TFS of 0 / 6. These results demonstrate that combination therapy is effective even when any one of the drugs is inactive. [Table 5]

[0270] In conclusion, the combination of IMGN632 and venetoclax demonstrated synergistic cytotoxicity in vitro, greater inhibition of tumor growth, and extended survival in vivo.

[0271] Example 5 In vivo efficacy of the combination of IMGN632 (QW x 3) and azacitidine (QD x 5) in a Molm-13 seeding model. To test the efficacy of IMGN632, azacitidine, or a combination of these two drugs in vivo for their ability to reduce tumor burden, disseminated tumor models were used as described in the protocol below.

[0272] Female NOD SCID mice were pre-treated with 150 mg / kg of cyclophosphamide, and partial bone marrow removal was performed to improve Molm-13 cell engraftment. Cyclophosphamide (Sigma, C0768, lot number MKBX1822V) was dissolved in 0.9% NaCl and administered intraperitoneally to the mice two days before Molm-13 cell inoculation on day 0.

[0273] Following the cyclophosphamide treatment described above, on day 0 of the study, 2 × 10¹⁶ units were added to 100 μl of serum-free medium. 5 One Molm-13 cell line (human AML cell line) was intravenously injected into the lateral tail vein of each mouse. Six days after Molm inoculation, the mice were randomized to test groups based on body weight. Twenty-four hours prior to each conjugate administration, mice in all groups receiving conjugate therapy (either alone or in combination with azacitidine) were intraperitoneally injected with 150 mg / kg of non-targeted chKTI antibody to block Fc receptors on Molm-13 AML cells and prevent nonspecific uptake of the conjugate.

[0274] Mice received IMGN632 once weekly (QW) in three doses (×3) at a dose of 0.3 μg / kg (0.024 mg / kg by DGN549, or 0.024 mg / kg by huCD123Ab), either alone or in combination with azacitidine, according to three different IMGN632 dosing schedules. These three different IMGN632 administration schedules were i) on days 7, 14, and 21 (referred to as "day 7" for IMGN632), ii) on days 11, 18, and 25 (referred to as "day 11" for IMGN632), and iii) on days 14, 21, and 28 (referred to as "day 14" for IMGN632), where the initiation date of IMGN632 treatment described was progressively moved away from day 0 of the study timeline (day of Molm-13 vaccination).

[0275] Azacitidine was administered once daily (QD) at a dose of 3.5 mg / kg for 5 consecutive days (×5; days 7, 8, 9, 10, and 11; "day 7" schedule), either alone or in combination with IMGN632.

[0276] Mice were treated with a combination regimen of IMGN632 (QW x 3) and 3.5 mg / kg azacitidine (QD x 5) according to one of three different dosing schedules for combining the two drugs: i) IMGN632 day 7 schedule and azacitidine day 7 schedule (Group F), ii) IMGN632 day 11 schedule and azacitidine day 7 schedule (Group G), and iii) IMGN632 day 14 schedule and azacitidine day 7 schedule (Group H).

[0277] The results are shown in Table 6 (below) and Figure 7.

[0278] Azacitidine monotherapy administered at 3.5 mg / kg in QD×5 according to the day 7 schedule (Group B) showed the lowest activity in this study, resulting in a 6-day tumor growth delay (TC value) and a 30% increase in ILS (life expectancy) compared to vehicle therapy. Three different treatment schedules for monotherapy IMGN632 showed either high activity ("Day 7" or "Day 11") or lowest activity ("Day 14") in this study, resulting in the following TC values ​​and %ILS: i) Day 7 schedule of IMGN632 (Group C) showed a TC of over 101 days and an ILS of over 505% (high activity); ii) Day 11 schedule of IMGN632 (Group D) showed a TC of 9.5 days and an ILS of 47.5% (high activity); iii) Day 14 schedule of IMGN632 (Group E) showed a TC of 6.5 days and an ILS of 32.5% (lowest activity).

[0279] The three different azacitidine + IMGN632 combination therapy regimens outlined above yielded the following antitumor activity: i) IMGN632 day 7 schedule + azacitidine combination (Group F) resulted in a TC value of 23.5 days and an ILS of 117.5% (high activity); ii) IMGN632 day 11 schedule + azacitidine combination (Group G) resulted in a TC value of 60.5 days and an ILS of 302.5% (high activity); iii) IMGN632 day 14 schedule + azacitidine combination (Group H) resulted in a TC value of 37 days and an ILS of 185% (high activity). Comparing all three combination schedules, Group G yielded the highest %ILS (ILS 302.5%), where IMGN632 treatment was initiated on the last azacitidine administration day (Day 11).

[0280] Notably, all three IMGN632 + azacitidine combination drug administration schedules tested yielded highly active combinations, but the corresponding IMGN632 monotherapy regimens (Group D, "Day 11": ILS 47.5%, and Group E, "Day 14": ILS) showed a high activity rate. Only the G group (ILS 302.5%) and H group (ILS 185%) schedules yielded a %ILS better than 32.5% (where the initiation of IMGN632 treatment was delayed compared to the initiation of azacitidine treatment). In contrast, the F group combination drug administration schedule (using the same initiation date for both IMGN632 and azacitidine, on day 7) resulted in an ILS of 117.5%, which was highly active but significantly lower than the corresponding IMGN632 monotherapy group (group C, "day 7"), with an ILS of over 505%. [Table 6]

[0281] Example 6 In vivo efficacy of IMGN632 (QW x 3) and azacitidine (QD x 5) combination in a Molm-13 disseminated tumor model. To test the efficacy of IMGN632, decitabine, or a combination of these two drugs in vivo for their ability to reduce tumor burden, a disseminated tumor model was used as described in the protocol below.

[0282] Female NOD SCID mice were pre-treated with 150 mg / kg of cyclophosphamide, and partial bone marrow removal was performed to improve Molm-13 cell engraftment. Cyclophosphamide (Sigma, C0768, lot number MKBX1822V) was dissolved in 0.9% NaCl and administered intraperitoneally to the mice two days before Molm-13 cell inoculation on day 0.

[0283] Following the cyclophosphamide treatment described above, on day 0 of the study, 2 × 10¹⁶ units were added to 100 μl of serum-free medium. 5 One Molm-13 cell line (human AML cell line) was intravenously injected into the lateral tail vein of each mouse. On day 5 after Molm inoculation, the mice were randomized to test groups based on body weight. 24 hours prior to each conjugate administration, mice in all groups receiving conjugate therapy (either alone or in combination with decitabine) were intraperitoneally injected with 150 mg / kg of non-targeted chKTI antibody to block Fc receptors on Molm-13 AML cells and prevent nonspecific uptake of the conjugate.

[0284] Mice received IMGN632 at a dose of 0.3 μg (0.024 mg / kg by huCD123Ab) once weekly (QW) in three doses (×3) according to three different IMGN632 dosing schedules, either alone or in combination with decitabine. These three different IMGN632 dosing schedules were i) on days 7, 14, and 21 (referred to as "day 7" for IMGN632), ii) on days 11, 18, and 25 (referred to as "day 11" for IMGN632), and iii) on days 14, 21, and 28 (referred to as "day 14" for IMGN632), where the initiation date of IMGN632 treatment described was progressively moved away from day 0 of the study timeline (day of Molm-13 vaccination).

[0285] Decitabine was administered once daily (QD) at a dose of 0.75 mg / kg for 5 consecutive days (×5; days 7, 8, 9, 10, and 11; "day 7" schedule), either alone or in combination with IMGN632.

[0286] Mice were treated with a combination regimen of IMGN632 (QW x 3) and 0.75 mg / kg decitabine (QD x 5) according to one of three different dosing schedules for combining the two drugs: i) IMGN632 day 7 schedule and decitabine day 7 schedule (Group F), ii) IMGN632 day 11 schedule and decitabine day 7 schedule (Group G), and iii) IMGN632 day 14 schedule and decitabine day 7 schedule (Group H).

[0287] The results are shown in Table 7 (below) and Figure 8.

[0288] Decitabine monotherapy administered at 0.75 mg / kg in QD×5 according to the day 7 schedule (Group B) showed the lowest activity in this study, resulting in a 6-day tumor growth delay (TC value) and a 28.6% increase in ILS (lifespan) compared to vehicle therapy. Three different treatment schedules for monotherapy IMGN632 were either highly active ("Day 7" or "Day 11") or inactive ("Day 14") in this study, resulting in the following TC values ​​and %ILS: i) Day 7 schedule of IMGN632 (Group C) had a TC of 49.5 days and an ILS of 235.7% (high activity); ii) Day 11 schedule of IMGN632 (Group D) had a TC of 20 days and an ILS of 95.2% (high activity); iii) Day 14 schedule of IMGN632 (Group E) had a TC of 0 days and an ILS of 0% (inactive).

[0289] The three different decitabine + IMGN632 combination therapy regimens outlined above yielded the following antitumor activity: i) IMGN632 day 7 schedule + decitabine combination (Group F) resulted in a TC value of 34 days and an ILS of 161.9% (high activity); ii) IMGN632 day 11 schedule + decitabine combination (Group G) resulted in a TC value of 38.5 days and an ILS of 183.3% (high activity); iii) IMGN632 day 14 schedule + decitabine combination (Group H) resulted in a TC value of 22 days and an ILS of 104.8% (high activity). Comparing all three combination schedules, the combination schedule that yielded the highest %ILS was Group G (ILS 183.3%), where IMGN632 treatment was initiated on the last day of decitabine administration (Day 11).

[0290] Notably, while all three IMGN632 + decitabine combination drug administration schedules tested resulted in highly active combinations, only schedules G (ILS 183.3%) and H (ILS 104.8%) yielded superior %ILS compared to the corresponding IMGN632 monotherapy regimens (Group D, "Day 11": 95.2% ILS, and Group E, "Day 14": 0% ILS, respectively) (where the initiation of IMGN632 treatment was delayed relative to the initiation of decitabine treatment). In contrast, the Group F combination drug administration schedule (using the same initiation date for both IMGN632 and decitabine, Day 7) yielded 161.9% ILS, which was highly active but significantly lower than the corresponding IMGN632 monotherapy group (Group C, "Day 7"), resulting in an ILS exceeding 235.7%. [Table 7]

[0291] Example 7 In vivo efficacy of IMGN632 (single dose) in combination with azacitidine (QD x 5) and venetoclax (QD x 28) in an EOL-1 subcutaneous model. To test the efficacy of a combination of IMGN632, azacitidine, and venetoclax, or a combination of these three drugs, in vivo for their ability to reduce tumor burden, a subcutaneous tumor model was used as described in the protocol below.

[0292] Each female athymic nude mouse was given 1 × 10⁶ mice in 100 μl of 50% Matrigel:50% serum-free medium (v / v). 7EOL-1 cells (human AML cell line) were subcutaneously inoculated into the right flank. On day 12, 24 hours before administration of the complex therapy (either alone or in combination with azacitidine + venetoclax), all mice in these therapy groups were intraperitoneally injected with 150 mg / kg of non-targeted chKTI antibody to block Fc receptors on EOL-1 AML cells and prevent nonspecific uptake of the complex. On day 12 after EOL-1 inoculation, mice were randomized to test groups based on tumor volume.

[0293] On day 13 after EOL-1 inoculation, mice received a single intravenous injection into the lateral tail vein of either the vehicle or IMGN632 at a dose of 1 μg / kg (by DGN549; 0.080 mg / kg by huCD123Ab). Venetoclax administration was also started on day 13, and mice receiving venetoclax received a single oral dose of 100 mg / kg of venetoclax daily (QD) for a total of 28 consecutive days (×28), including the day after cell transplantation, from day 13 to day 40. Azacitidine administration was also started on day 13, and mice receiving azacitidine received a single intraperitoneal dose of 3 mg / kg of azacitidine daily (QD) for 5 consecutive days (×5). In the combination groups, mice received either IMGN632 and either venetoclax plus azacitidine, or azacitidine alone, as outlined above.

[0294] The results are shown in Table 8 and Figure 9.

[0295] A single dose of IMGN632 monotherapy at 1 μg / kg resulted in a T / C ratio of 64% (inactive) and 2 / 6 long-term complete regression (tumor-free survivors, TFS) at the end of the study (day 77). A monotherapy regimen of azacitidine 3 mg / kg once daily (QD) for 5 days (×5) was inactive, with a T / C ratio of 96% and a TFS of 0 / 6 at day 77. The combination therapy of azacitidine (QD × 5) and venetoclax (QD × 28) was inactive, with a T / C ratio of 46% and a TFS of 1 / 6 at day 77.

[0296] The triple therapy of IMGN632, azacitidine, and venetoclax was highly effective, with a T / C ratio of 0% and a TFS of 5 / 6 at the end of the study. The triple therapy resulted in a lower %T / C ratio (0%) than either IMGN632 monotherapy (64%) or azacitidine + venetoclax dual therapy (46%), indicating that the triple therapy was more effective in slowing tumor growth than IMGN632 monotherapy or azacitidine + venetoclax dual therapy.

[0297] The combination therapy of IMGN632 and azacitidine also showed high activity, with a T / C ratio of 0% and a TFS of 4 / 6 at the end of the study. The combination therapy of IMGN632 and azacitidine resulted in a lower %T / C ratio (0%) than that achieved by either IMGN632 monotherapy (64%) or azacitidine monotherapy (96%), indicating that the combination therapy was more effective in slowing tumor growth than the corresponding monotherapy. [Table 8]

[0298] Example 8 In vivo efficacy of the combination of IMGN632 (QW x 3), azacitidine (QD x 5), and venetoclax (QD x 28) in a Molm-13 seeding model. To test the efficacy of IMGN632, venetoclax, azacitidine + venetoclax combination, IMGN632 + venetoclax combination, or a triple combination of IMGN632, azacitidine, and venetoclax in vivo in reducing tumor burden, disseminated tumor models were used as described in the following protocols.

[0299] Female NOD SCID mice were pre-treated with 150 mg / kg of cyclophosphamide, and partial bone marrow removal was performed to improve Molm-13 cell engraftment. Cyclophosphamide (Sigma, C0768, lot number MKBX1822V) was dissolved in 0.9% NaCl and administered intraperitoneally to the mice two days before Molm-13 cell inoculation on day 0.

[0300] Following the cyclophosphamide treatment described above, on day 0 of the study, 2 × 10¹⁶ units were added to 100 μl of serum-free medium. 5 One Molm-13 cell line (human AML cell line) was intravenously injected into the lateral tail vein of each mouse. Twenty-four hours prior to each conjugate administration, mice in all groups receiving conjugate therapy (either alone, in combination with venetoclax, or in combination with azacitidine + venetoclax) were intraperitoneally injected with 150 mg / kg of non-targeted chKTI antibody to block Fc receptors on Molm-13 AML cells and prevent nonspecific uptake of the conjugate.

[0301] On days 11, 18, and 25 after Molm-13 inoculation, mice received a single intravenous injection into the lateral tail vein of either vehicle or IMGN632 at 0.3 μg / kg (by DGN549; 0.024 mg / kg by huCD123Ab). Venetoclax administration was initiated on day 7 after cell inoculation, and mice receiving venetoclax received a single oral dose of 100 mg / kg of venetoclax daily (QD) for a total of 28 consecutive days (×28) from days 7 to 34 (including days 7 and 34). Azacitidine administration was also initiated on day 7, and mice receiving azacitidine received a single intraperitoneal dose of 3.5 mg / kg of azacitidine daily (QD) for 5 consecutive days (×5). In the groups receiving azacitidine and venetoclax in combination or in the group receiving all three drugs in triple combination, mice were administered IMGN632, venetoclax, and azacitidine, as outlined above.

[0302] The results up to 80 days after tumor cell inoculation are shown in Table 9A (below) and Figure 10A.

[0303] IMGN632 monotherapy (Group B) resulted in a tumor growth delay of over 59.5 days (TC value) and an increase in lifespan of over 277% (%ILS) compared to vehicle therapy. Venetoclax monotherapy (Group E) resulted in a TC of 11.5 days and an ILS of 53.5% (high activity). Azacitidine and venetoclax combination therapy (Group C) resulted in a TC of 14.5 days and an ILS of 67.4% (high activity). IMGN632 and venetoclax combination therapy (Group F) resulted in a TC of over 59.5 days and an ILS of over 277% (high activity) (higher TC and %ILS values ​​than those achieved with venetoclax monotherapy), indicating that the addition of IMGN632 to venetoclax provides additional benefits compared to venetoclax monotherapy. The triple therapy combination of IMGN632, azacitidine, and venetoclax (Group D) resulted in a total therapeutic interval (TC) of over 59.5 days and an intracellular salping (ILS) of over 277% (high activity) (higher TC and %ILS values ​​than those achieved with the dual therapy of azacitidine and venetoclax), indicating that the addition of IMGN632 to the dual therapy of azacitidine and venetoclax provides additional benefits compared to treatment with azacitidine and venetoclax alone. [Table 9A]

[0304] The results up to 109 days after tumor cell inoculation are shown in Table 9B (below) and Figure 10B. Compared to vehicle therapy, IMGN632 monotherapy (Group B) resulted in a tumor growth delay of over 87.5 days (TC value) and an increase in lifespan of over 407% (%ILS). Venetoclax monotherapy (Group E) resulted in a TC of 11.5 days and an ILS of 53.5% (high activity). Azacitidine and venetoclax combination therapy (Group C) resulted in a TC of 14.5 days and an ILS of 67.4% (high activity). The combination therapy of IMGN632 and venetoclax (Group F) resulted in a TC of over 87.5 days and an ILS of over 407% (high activity) (higher TC and %ILS values ​​than those achieved with venetoclax alone), indicating that the addition of IMGN632 to venetoclax provides additional benefits compared to venetoclax monotherapy. The tripartite combination therapy of IMGN632, azacitidine, and venetoclax (Group D) also resulted in a TC of over 87.5 days and an ILS of over 407% (high activity) (higher TC and %ILS values ​​than those achieved with the combination therapy of azacitidine and venetoclax), indicating that the addition of IMGN632 to the combination therapy of azacitidine and venetoclax provides additional benefits compared to treatment with azacitidine and venetoclax alone. [Table 9B]

[0305] Example 9 In vivo efficacy of the combination of IMGN632 (QW x 3), azacitidine (QD x 5), and venetoclax (QD x 28) in an MV4-11 seeding model. To test the efficacy of a combination of IMGN632, azacitidine, and venetoclax, or a combination of these three drugs, in vivo for their ability to reduce tumor burden, disseminated tumor models were used as described in the protocol below.

[0306] Female NOD SCID mice were pre-treated with 150 mg / kg of cyclophosphamide, and the bone marrow was partially removed to improve MV4-11 cell engraftment. Cyclophosphamide (Sigma, C0768, lot number MKBX1822V) was dissolved in 0.9% NaCl and administered intraperitoneally to mice on day 0, 3 days and 2 days before MV4-11 cell inoculation.

[0307] Following the cyclophosphamide treatment described above, on day 0 of the study, 3 × 10¹⁶ units were added to 100 μl of serum-free medium. 6 Each mouse was intravenously injected with MV4-11 cells (human AML cell line) into the lateral tail vein. Three days after MV4-11 inoculation, the mice were randomized to test groups based on body weight. Twenty-four hours prior to each conjugate administration, mice in all groups receiving conjugate therapy (either alone or in combination with azacitidine and venetoclax) were intraperitoneally injected with 150 mg / kg of non-targeted chKTI antibody to block Fc receptors on MV4-11 AML cells and prevent nonspecific uptake of the conjugate.

[0308] On days 25, 32, and 39 after MV4-11 inoculation, mice received a single intravenous injection of either vehicle or IMGN632 at a dose of 1 μg / kg (by DGN549; 0.080 mg / kg by huCD123Ab) into the lateral tail vein. Venetoclax administration was initiated on day 21 after cell inoculation, and mice receiving venetoclax received a single oral dose of 100 mg / kg of venetoclax daily (QD) for a total of 28 consecutive days (×28) from days 21 to 48 (including days 21 and 48). Azacitidine administration was also initiated on day 21, and mice receiving azacitidine received a single intraperitoneal dose of 4.5 mg / kg or 3 mg / kg of azacitidine daily (QD) for 5 consecutive days (×5). In the azacitidine + venetoclax combination group or the triple combination group of the three drugs, mice received IMGN632, venetoclax, and azacitidine, as outlined above.

[0309] The results are shown in Table 10 and Figure 11.

[0310] IMGN632 monotherapy (Group B) resulted in a tumor growth delay of over 81 days (TC value) and an increase in lifespan of over 180% compared to vehicle therapy. Dual therapy with azacitidine (4.5 mg / kg) and venetoclax (Group C) resulted in a TC of 28 days and an ILS of 62.2% (high activity). Triple therapy with IMGN632, azacitidine (4.5 mg / kg), and venetoclax (Group D) resulted in a TC of 64 days and an ILS of over 142.2% (high activity) (higher TC and %ILS values ​​than those achieved by dual therapy with azacitidine (4.5 mg / kg) and venetoclax), indicating that the addition of IMGN632 to dual therapy with azacitidine and venetoclax provides additional benefits compared to treatment with azacitidine and venetoclax alone.

[0311] In the dual therapy of azacitidine (3 mg / kg) and venetoclax (Group E), the total therapeutic interval (TC) was 20 days and the intracellular salping (ILS) was 44.4% (active). In the tripartite therapy of IMGN632, azacitidine (3 mg / kg), and venetoclax (Group D), the TC was 76 days and the ILS was 168.9% (highly active) (higher TC and %ILS values ​​than those achieved by the dual therapy of azacitidine (3 mg / kg) and venetoclax), indicating that the addition of IMGN632 to the dual therapy of azacitidine and venetoclax provides additional benefits compared to treatment with azacitidine and venetoclax alone. [Table 10]

[0312] Example 10: Administration of IMGN632 combinations to humans Patients with incompatible frontline and relapsed / refractory BPDCN and relapsed / refractory ALL are treated with IMGN632 monotherapy at the recommended phase 2 dose (RP2D) of 0.045 mg / kg once every 21-day cycle (Q3W). Based on the above experiments, the following three dosing schedules were selected for administration to human subjects.

[0313] Schedule A: IMGN632 + Azacitidine: The cycle for IMGN632 and azacitidine is 28 days. In this 28-day cycle, azacitidine is administered daily at 75 mg / m² on days 1-7. 2 It is administered subcutaneously (SC) or intravenously (IV). Alternatively, azacitidine is administered daily at 75 mg / m² for days 1-5. 2 It can be administered subcutaneously (SC) or intravenously (IV). Administration of IMGN632 is initiated on day 7 of the last azacitidine dose. IMGN632 is administered in total doses of 0.015 mg / kg, 0.045 mg / kg, 0.09 mg / kg, or 0.18 mg / kg per 28-day cycle. For example, IMGN632 is administered once every 28-day cycle (i.e., on day 7) in doses of 0.015 mg / kg, 0.045 mg / kg, or 0.09 mg / kg. IMGN632 may also be administered once every 28-day cycle (e.g., on day 7) in doses of 0.03 mg / kg. Alternatively, IMGN632 may be administered three times every 28-day cycle (i.e., on days 7, 10, and 14) at doses of 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg.

[0314] If, after cycle 1, the patient experiences toxicity associated with azacitidine (e.g., cytopenia), azacitidine administration may be reduced to 5 days by omitting treatment on days 1 and 2 and administering azacitidine on days 3–7 in subsequent cycles.

[0315] A total of 2 to 12 cycles of administration are possible. Subsequently, IMGN632 may be administered as maintenance therapy once every three weeks (for example, at a dose of 0.015 mg / kg, 0.045 mg / kg, or 0.09 mg / kg on day 1 of a 21-day cycle) or three times every three weeks (for example, at a dose of 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg on days 1, 4, and 8 of a 21-day cycle). [Table 11-1] [Table 11-2]

[0316] Schedule B: IMGN632 + Venetocracy The IMGN632 and venetoclax cycle is 21 days. In the first cycle, venetoclax is administered orally (PO) at a dose of 100 mg on day 1, 200 mg on day 2, and 400 mg on all subsequent days (days 3-21). In subsequent cycles, venetoclax is administered at the maximum dose (e.g., 400 mg) on ​​all days. Administration of IMGN632 is initiated on day 7 after the seventh dose of venetoclax. IMGN632 is administered at a total dose of 0.015 mg / kg, 0.045 mg / kg, 0.09 mg / kg, or 0.18 mg / kg per 21-day cycle. For example, IMGN632 is administered once per 21-day cycle (e.g., on day 7) at a dose of 0.015 mg / kg, 0.045 mg / kg, or 0.09 mg / kg. IMGN632 may also be administered once every 28-day cycle (e.g., on day 7) at a dose of 0.03 mg / kg. Alternatively, IMGN632 may be administered three times every 21-day cycle (i.e., on days 7, 10, and 14) at doses of 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg.

[0317] According to the FDA labeling for venetoclax, potent cytochrome P450 (CYP) 3A inhibitors, such as voriconazole and posaconazole, should be avoided. If a patient is taking a moderate CYP3A inhibitor (e.g., Cresemba), the maximum venetoclax dose should be 200 mg.

[0318] If, after Cycle 1, the patient experiences venetoclax-associated toxicity (e.g., cytopenia), venetoclax dosage may be reduced between days 1 and 14 (or between days 1 and 7). Venetoclax dosage may also be reduced between days 1 and 8.

[0319] A total of 2 to 12 cycles of administration are possible. Subsequently, IMGN632 may be administered as maintenance therapy once every three weeks (for example, at a dose of 0.015 mg / kg, 0.045 mg / kg, or 0.09 mg / kg on day 1 of a 21-day cycle) or three times every three weeks (for example, at a dose of 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg on days 1, 4, and 8 of a 21-day cycle). [Table 12-1] [Table 12-2]

[0320] Schedule C: IMGN632 + Azacitidine + Venetoclax The cycle for IMGN632, azacitidine, and venetoclax is 28 days. In this 28-day cycle, azacitidine is administered daily at 75 mg / m² on days 1-7. 2 It is administered subcutaneously (SC) or intravenously (IV). Alternatively, azacitidine is administered daily at 75 mg / m² for days 1-5. 2 It can be administered subcutaneously (SC) or intravenously (IV).

[0321] In the first cycle, venetoclax is administered orally (PO) at a dose of 100 mg on day 1, 200 mg on day 2, and 400 mg on all subsequent days (days 3-28). In subsequent cycles, venetoclax is administered orally (PO) at the maximum dose in the first cycle (e.g., 400 mg on all days). Alternatively, venetoclax can be administered at a dose of 100 mg on day 1, 200 mg on day 2, and 400 mg on days 3-8, 3-14, or 3-21 of a 28-day cycle. In subsequent cycles, the maximum dose (e.g., 400 mg) can be administered orally (PO) at days 1-8, 1-14, 1-18, or 1-21 of a 28-day cycle.

[0322] IMGN632 administration is initiated on day 7, for example, after the 7th dose of venetoclax and azacitidine. IMGN632 is administered in a total dose of 0.015 mg / kg, 0.045 mg / kg, 0.09 mg / kg, or 0.18 mg / kg per 28-day cycle. For example, IMGN632 may be administered once per 28-day cycle (e.g., on day 7) in doses of 0.015 mg / kg, 0.045 mg / kg, or 0.09 mg / kg. IMGN632 may also be administered once per 28-day cycle (e.g., on day 7) in doses of 0.03 mg / kg. Alternatively, IMGN632 may be administered three times per 28-day cycle (i.e., on days 7, 10, and 14) in doses of 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg.

[0323] A total of 2 to 12 cycles of administration are possible. Subsequently, IMGN632 may be administered as maintenance therapy once every three weeks (for example, at a dose of 0.015 mg / kg, 0.045 mg / kg, or 0.09 mg / kg on day 1 of a 21-day cycle) or three times every three weeks (for example, at a dose of 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg on days 1, 4, and 8 of a 21-day cycle). IMGN632 can also be administered as maintenance therapy once every three weeks (for example, at a dose of 0.03 mg / kg on day 1 of a 21-day cycle). [Table 13-1] [Table 13-2]

[0324] Example 11: Administration to IMGN632 human patients with minimal residual disease The IMGN632 study was designed to evaluate the efficacy of intravenous administration of IMGN632 in adult leukemia patients with minimal residual disease (MRD). Patients included both eligible and ineligible individuals. Patients with MRD may have a lower leukemia burden and therefore lower CD123 levels than other patients. Thus, a higher IMGN632 to CD123 ratio may exist in these patients compared to other patients, potentially allowing for lower doses (e.g., 15–45 mcg / kg) to be effective.

[0325] Patients enrolled in this study must be in complete remission (CR / CRi), be MRD+ after intensive induction / intensification therapy, and have no access to appropriate standard treatment. MRD status will be assessed using a central flow cytometry-based study.

[0326] Patients may receive a premedication regimen before each IMGN632 infusion. Premedication may include (i) 25–50 mg of diphenhydramine (IV or oral [PO]), (ii) 325–650 mg of acetaminophen or paracetamol (IV or PO), and / or (iii) 8 mg of dexamethasone (PO or IV). If an individual patient requires more intensive or alternative treatment to prevent an infusion reaction (e.g., different corticosteroids, different doses of any medication), the regimen may be modified according to standard institutional practice.

[0327] IMGN632 is administered once every three weeks (21 days) at doses of 0.015 mg / kg, 0.045 mg / kg, 0.09 mg / kg, 0.135 mg / kg, or 0.18 mg / kg.

[0328] The treatment consists of two cycles (i.e., a total of 6 weeks), with the patient's second dose administered at least 21 days after the first dose. For example, patients who benefit from this regimen may receive up to 10 or more additional cycles in total.

[0329] Antileukemic activity is demonstrated by measuring the conversion rate from MRD+ to MRD-. Relapse-free survival (RFS) and event-free survival (EFS) are also evaluated to indicate antitumor activity.

[0330] Exemplary examples provided herein In one example provided herein (I1), a method for treating hematological malignancies in a subject comprises administering to the subject in need an immune complex that binds to CD123 (the immune complex includes an antibody or an antigen-binding fragment thereof, which includes a heavy chain variable region CDR1 containing the amino acid sequence of SEQ ID NO: 5, a heavy chain variable region CDR2 containing the amino acid sequence of SEQ ID NO: 6, and a heavy chain variable region CDR3 containing the amino acid sequence of SEQ ID NO: 7, as well as a light chain variable region CDR1 containing the amino acid sequence of SEQ ID NO: 8, a light chain variable region CDR2 containing the amino acid sequence of SEQ ID NO: 9, and a light chain variable region CDR3 containing the amino acid sequence of SEQ ID NO: 10), a BCL-2 inhibitor, a hypomethylating agent, or a combination thereof.

[0331] In one case of I1 (I2), the immune complex is administered in combination with a BCL-2 inhibitor. In one case of I1 (I3), the immune complex is administered in combination with a hypomethylating agent. In one case of I1 (I4), the immune complex is administered in combination with a BCL-2 inhibitor and a hypomethylating agent.

[0332] In one of the cases I1 to I4 (I5), the antibody or antigen-binding fragment includes a VH containing the amino acid sequence shown in SEQ ID NO: 1, and / or a VL containing the amino acid sequence shown in SEQ ID NO: 2. In one of the cases I5 (I6), the antibody or antigen-binding fragment includes a heavy chain constant region and / or a light chain constant region.

[0333] In one of the cases I1 to I4 (I7), the antibody or antigen-binding fragment includes a heavy chain containing the amino acid sequence shown in SEQ ID NO: 3, and / or a light chain containing the amino acid sequence shown in SEQ ID NO: 4.

[0334] In one of the cases I1-I7 (I8), the immune complex contains a cytotoxin, and optionally, the cytotoxin is a DNA alkylating agent. In one of the cases I8 (I9), the DNA alkylating agent is an indolino-benzodiazepine (IGN) DNA alkylating agent.

[0335] In one of the cases I1-I9 (I10), the immune complex contains a peptide linker.

[0336] In one of the cases I1 to I10 (I11), the immune complex is IMGN632.

[0337] In one of the cases I1 to I0 (I12), the immune complex was administered in a pharmaceutical composition containing an immune complex having the following structure: [ka] , in the formula, G4723A contains a heavy chain with the amino acid sequence shown in SEQ ID NO: 3 and a light chain with the amino acid sequence shown in SEQ ID NO: 4.

[0338] In one of the cases I1 to I12 (I13), the administration is considered first-line therapy.

[0339] In one of the cases I1-I13 (I14), the immune complex is administered intravenously.

[0340] In one of the cases (15) from I1 to I14, administration of an immune complex with a BCL-2 inhibitor, a hypomethylating agent, or a combination thereof produced a synergistic effect.

[0341] In one of the cases I1, I2, and any one of I4-I15 (I16), administration of immune complexes and BCL-2 inhibitors does not produce more toxicity than administration of immune complexes alone or BCL-2 inhibitors alone. In one of the cases I1 and any one of I3-I15 (I17), administration of immune complexes and hypomethylating agents does not produce more toxicity than administration of immune complexes alone or hypomethylating agents alone. In one of the cases I1 and any one of I4-I15 (I18), administration of immune complexes, BCL-2 inhibitors, and hypomethylating agents does not produce more toxicity than administration of immune complexes, BCL-2 inhibitors, and / or hypomethylating agents.

[0342] In cases I1, I2, and any one of I5-I18 (I19), the immune complex is administered once in a 21-day cycle. In case I19 (I20), the immune complex is administered once in a 21-day cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg, with the dose being optional, approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg.

[0343] In one case (I21) from I1 to I18, the immune complex is administered three times in a 21-day cycle. In one case (I22) from I21, the total amount of immune complex administered in the 21-day cycle is approximately 0.045 mg / kg, 0.09 mg / kg, or 0.18 mg / kg. In one case (I23) from I22, an immune complex of approximately 0.015 mg / kg to 0.06 mg / kg is administered in each of the three doses during the 21-day cycle, with optional doses of approximately 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg being administered in each of the three doses during the 21-day cycle. In one case (I24) from I21 to I23, the first administration of the immune complex is on day 7 of the 21-day cycle. In one of the cases I21-I24 (I25), the second dose of the immune complex was administered on day 10 of the 21-day cycle. In one of the cases I21-I25 (I26), the third dose of the immune complex was administered on day 14 of the 21-day cycle. In one of the cases I21-I26 (I27), the first, second, and third doses were administered on days 7, 10, and 14 of the 21-day cycle, respectively.

[0344] In one of the cases I1, I2, and I4-I27 (I28), the BCL-2 inhibitor was venetoclax.

[0345] In cases I1, I2, and any one of I4-I28 (I29), the BCL-2 inhibitor is administered at a dose of 400 mg. In case I1, I2, and any one of I4-I28 (I30), the BCL-2 inhibitor is administered at a dose of 200 mg.

[0346] In one case (I31) of I1, I2, and I4-I30, the BCL-2 inhibitor is administered daily in a 21-day cycle. In one case (I32) of I1, I2, and I4-I28, the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 21-day cycle, 200 mg on day 2 of the 21-day cycle, and 400 mg on days 3-21 of the 21-day cycle. In one case (I33) of I1, I2, and I4-I28, the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 21-day cycle, 200 mg on day 2 of the 21-day cycle, and 400 mg on days 3-7 or 3-14 of the 21-day cycle. In cases I1, I2, and any one of I4-I28 (I34), the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 21-day cycle, and at a dose of 200 mg on days 2-21, 2-14, or 2-7 of the 21-day cycle.

[0347] In cases I1, I2, and one of the cases I4-I34 (I35), the BCL-2 inhibitor is administered orally.

[0348] In cases I1, I2, and any one of I4-I35 (I36), immune complex administration is initiated 6 days after the start of BCL-2 inhibitor administration.

[0349] In one case (I37) from I1 and I3-I18, the immune complex is administered once in a 28-day cycle. In the case (I38) from I37, the immune complex is administered once in a 28-day cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg, with the dose being optional, approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg.

[0350] In one case (I39) from I1 to I18, the immune complex is administered three times in a 28-day cycle. In one case (I40) of I39, the total amount of immune complex administered in the 28-day cycle is approximately 0.045 mg / kg, approximately 0.09 mg / kg, or approximately 0.18 mg / kg. In one case (I41) of I40, an immune complex of approximately 0.015 mg / kg to approximately 0.06 mg / kg is administered in each of the three doses during the 28-day cycle, with optional doses of approximately 0.015 mg / kg, approximately 0.03 mg / kg, or approximately 0.06 mg / kg being administered in each of the three doses during the 28-day cycle. In one case (I42) from I39 to I41, the first administration of the immune complex is on day 7 of the 28-day cycle. In one of the cases I39-I42 (I43), the second dose of the immune complex was administered on day 10 of the 28-day cycle. In one of the cases I39-I43 (I44), the third dose of the immune complex was administered on day 14 of the 28-day cycle. In one of the cases I39-I44 (I45), the first, second, and third doses were administered on days 7, 10, and 14 of the 28-day cycle, respectively.

[0351] In one of the cases I37-I45 (I46), the BCL-2 inhibitor is venetoclax. In one of the cases I37-I46 (I47), the BCL-2 inhibitor is administered at a dose of 400 mg. In one of the cases I37-I46 (I48), the BCL-2 inhibitor is administered at a dose of 200 mg. In one of the cases I37-I48 (I49), the BCL-2 inhibitor is administered daily in a 28-day cycle. In one of the cases I37-I46 (I50), the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 28-day cycle, at a dose of 200 mg on day 2 of the 28-day cycle, and at a dose of 400 mg on days 3-28 of the 28-day cycle. In one of the cases I37-I46 (I51), the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 28-day cycle, 200 mg on day 2 of the 28-day cycle, and 400 mg on days 3-7 or 3-14 of the 28-day cycle. In one of the cases I37-I46 (I52), the BCL-2 inhibitor is administered at a dose of 100 mg on day 1 of the 28-day cycle, and 200 mg on days 2-28, 2-14, or 2-7 of the 28-day cycle. In one of the cases I37-I52 (I53), the BCL-2 inhibitor is administered orally. In one of the cases I37-I53 (I54), the administration of immune complexes is initiated 6 days after the start of BCL-2 inhibitor administration.

[0352] In one case (I55) of I1, I3-I18, and I37-I54, the hypomethylating agent is azacitidine. In one case (I56) of I55, azacitidine is administered in a 28-day cycle. In one case (I57) of I55 or I56, azacitidine is administered once daily on days 1-7 of the 28-day cycle. In one case (I58) of I55 or I56, azacitidine is administered once daily on days 3-7 of the 28-day cycle. In one case (I59) of I56-I58, the azacitidine dose is approximately 75 mg / m². 2It is administered in the following dose. In one of the cases I56-I59 (I60), azacitidine is administered subcutaneously or intravenously.

[0353] In one of the cases I1 and I3-I54 (I61), the hypomethylating agent is decitabine. In the case of I61 (I62), decitabine is administered intravenously.

[0354] In one of the cases I1 to I62 (I63), hematological malignancies are present as minimal residual disease (MRD).

[0355] In one case (I64), the treatment method for hematological malignancies present as minimal residual disease in human subjects includes administering anti-CD123 immune complexes containing anti-CD123 antibodies or antigen-binding fragments linked to cytotoxic agents to the target. In one case of I64 (I65), the immune complexes are administered at doses of approximately 0.045 mg / kg to approximately 0.18 mg / kg. In one case of I64 or I65 (I66), the target is administered approximately 0.045 mg / kg, approximately 0.09 mg / kg, approximately 0.135 mg / kg, or approximately 0.18 mg / kg. In any one case from I64 to I66 (I67), the immune complexes are administered intravenously. In any one case from I64 to I67 (I68), the hematological malignancy is leukemia. In one of the cases I64-I68 (I69), the immune complex is administered to the subject once in a 21-day cycle.

[0356] In one case (I70) from I20-I63 or I69, the administration is 1 cycle. In one case (I71) from I20-I63 or I69, the administration is 2 or more cycles. In one case (I72) from I20-I63 or I69, the administration is at least 2 cycles, at least 3 cycles, at least 4 cycles, at least 5 cycles, at least 6 cycles, at least 7 cycles, at least 8 cycles, at least 9 cycles, or at least 10 cycles. In one case (I73) from I20-I63 or I69, the administration is approximately 2-4 cycles, approximately 2-6 cycles, approximately 2-8 cycles, approximately 2-10 cycles, or approximately 2-12 cycles.

[0357] In any one case among I1 to I73 (I74), the method further includes administering a reduced dose of the immune complex after dose-limiting toxicity has occurred in the subject and has been reduced to baseline or grade 2 or lower.

[0358] In one of the cases I1-I73 (I75), the immune complex is administered further as maintenance therapy. In one case of I75 (I76), maintenance therapy includes one administration of the immune complex every 21 days. In one case of I76 (I77), maintenance therapy includes one administration of the immune complex every 21 days at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg, with the dose being optional, approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg. In one case of I75 (I78), maintenance therapy includes three administrations of the immune complex every 21 days. In one case of I78 (I79), the total amount of immune complex administered every 21 days during maintenance therapy is approximately 0.045 mg / kg, approximately 0.09 mg / kg, or approximately 0.18 mg / kg. In one case of I79 (I80), during maintenance therapy, approximately 0.015 mg / kg to approximately 0.06 mg / kg of immune complexes were administered on each of the three 21-day cycles, with optional administration of approximately 0.015 mg / kg, approximately 0.03 mg / kg, or approximately 0.06 mg / kg of immune complexes on each of the three 21-day cycles. In one case of any one of I78-I80 (I81), during maintenance therapy, the first, second, and third doses were administered on days 1, 4, and 8 of the 21-day cycle, respectively.

[0359] In one of the cases (I82) among I1-I12, I14-I62, and I69-I81, the hematological malignancy is a recurrent hematological malignancy.

[0360] In one of the cases (I83) from I1 to I82, the hematological malignancies were acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia in acute transformation / acute transformation phase (BP-CML), and blastic plasmacytoid dendritic cell neoplasm (BPDCN).

[0361] In one case from I1 to I82 (I84), the hematological malignancy is AML. In one case from I84 (I85), the AML is relapsed AML. In one case from either I84 or I85 (I86), the AML is refractory AML.

[0362] In one case (I87) from I1 to I82, the hematological malignancy is BPDCN. In one case (I88) from I88, the BPDCN is recurrent BPDCN. In one case (I89) from either I87 or I88, the BPDCN is refractory BPDCN.

[0363] In one case (I90) from I1 to I82, the hematological malignancy is ALL. In the case of I90 (I91), the ALL is recurrent ALL. In the case of either I90 or I91 (I92), the ALL is refractory ALL.

[0364] In one of the cases from I1 to I82 (I93), the hematological malignancy is chronic myelomonocytic leukemia (CMML). In one of the cases from I93 (I94), the CMML is relapsed CMML. In one of the cases from I93 or I94 (I95), the CMML is refractory CMML.

[0365] In one case from I1 to I82 (I96), the hematological malignancy is myelofibrosis (MF). In one case from I96 (I97), the MF is relapsed MF. In one case from either I96 or I97 (I98), the MF is refractory MF.

[0366] In one case (I99) from I1 to I82, the hematological malignancy is myelodysplastic syndrome (MDS). In the case of I99 (I100), the MDS is relapsed MDS. In the case of either I99 or I100 (I101), the MDS is refractory MDS.

[0367] In one of the cases I1 to I101 (I102), the hematological malignancy is a hematological malignancy that expresses CD123.

[0368] In one of the cases I1-I102 (I103), CD123 was detected in a sample obtained from a hematological malignancy before administration. In another case of I103 (I104), CD123 was detected using flow cytometry.

[0369] In one of the cases I1 to I105 (I105), the method further includes detecting CD123 in a sample obtained from a hematological malignancy before administration.

[0370] In one of the cases I1-I105 (I106), at least 80% of cells in hematological malignancies express CD123. In one of the cases I1-I106 (I107), CD123 is detected in at least 80% of cells in samples obtained from hematological malignancies before administration.

[0371] In one of the cases I1 to I107 (I108), the method further includes detecting CD123 in at least 80% of cells in a sample obtained from a hematological malignancy before administration.

[0372] In one of the cases I1-I63 and I69-I108 (I109), the hematological malignancies were resistant to IMGN632.

[0373] In one of the cases I1-I109 (I110), the hematological malignancy exhibits multidrug resistance 1 (MDR1). In one of the cases I1-I110 (I111), the hematological malignancy expresses P-glycoprotein (P-gp). In one of the cases I1-I111 (I112), the subject has an absolute neutrophil count greater than 500 / μL.

[0374] In one case (I113) from among I1-I12 and I14-I112, the subject had received at least one prior therapy. In one case (I114) from among I1-I12 and I14-I112, the subject had received at least two prior therapies. In one case (I115) from among I1-I12 and I14-I112, the subject had received at least three prior therapies.

[0375] In one of the cases I1-I12 and I14-I115 (I116), the cancer had been previously treated with venetoclax. In one of the cases I1-I115 (I117), the cancer had not been previously treated with venetoclax.

[0376] In one of the cases I1-I12 and I14-I117 (I118), the cancer had been previously treated with a hypomethylating agent. In one of the cases I1-I117 (I119), the cancer had not been previously treated with a hypomethylating agent.

[0377] In one of the cases I1-I119 (I120), the subject was pre-treated with a corticosteroid prior to the administration of the immune complex. In one of the cases I1-I119 (I121), the method further includes pre-treating the subject with a corticosteroid prior to the administration of the immune complex. In one of the cases I120 or I121 (I122), the corticosteroid is diphenhydramine, acetaminophen, paracetamol, dexamethasone, or a combination thereof.

[0378] In one of the cases I1-I63 and I69-I122 (I123), the immune complex and the BCL-2 inhibitor, hypomethylating agent, or a combination thereof are administered in separate pharmaceutical compositions.

[0379] In one of the cases I1 to I123 (I124), the subject is a human.

[0380] In one case (I125), the treatment method for hematological malignancies in human subjects included administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously on day 7 of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (optional, with doses of approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg), and venetoclax administered orally at a dose of 100 mg on day 1 of the cycle, 200 mg on day 2 of the cycle, and 400 mg on days 3 to 21 of the cycle.

[0381] In one case (I126), the treatment method for hematological malignancies in human subjects involved administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously on day 7 of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (optional, with doses of approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg), and venetoclax administered orally at a daily dose of 400 mg.

[0382] In one case (I127), the treatment method for hematological malignancies in human subjects included administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 administered intravenously at doses of approximately 0.015 mg / kg to approximately 0.06 mg / kg on days 7, 10, and 14 of the cycle, and optionally administered at doses of approximately 0.015 mg / kg, approximately 0.03 mg / kg, or approximately 0.06 mg / kg on each day, and venetoclax administered orally at a dose of 100 mg on day 1 of the cycle, a dose of 200 mg on day 2 of the cycle, and a dose of 400 mg on days 3 to 21 of the cycle.

[0383] In one case (I128), the treatment method for hematological malignancies in human subjects involved administering IMGN632 and venetoclax in a 21-day cycle to subjects in need, with IMGN632 doses of approximately 0.015 mg / kg to approximately 0.06 mg / kg administered intravenously on days 7, 10, and 14 of the cycle, and optionally approximately 0.015 mg / kg, approximately 0.03 mg / kg, or approximately 0.06 mg / kg administered on each day, and venetoclax administered orally at a daily dose of 400 mg.

[0384] In one case (I129), the treatment method for hematological malignancies in human subjects involved administering IMGN632 and azacitidine in a 28-day cycle to subjects in need, with IMGN632 administered intravenously on day 7 of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg (optional, with doses of approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg), and azacitidine administered at 75 mg / m² on days 1 to 7 of the cycle. 2 It is administered subcutaneously or intravenously in the specified dose.

[0385] In one case study (I130), the treatment method for hematological malignancies in human subjects involved administering IMGN632 and azacitidine in a 28-day cycle to subjects in need, with approximately 0.015 mg / kg to 0.06 mg / kg of IMGN632 administered intravenously on days 7, 10, and 14 of the cycle (optionally, doses of approximately 0.015 mg / kg, 0.03 mg / kg, or 0.06 mg / kg administered on each day), and 75 mg / m² of azacitidine administered on days 1 to 7 of the cycle. 2 It is administered subcutaneously or intravenously in the specified dose.

[0386] In one case of I129 or I130 (I131), the method further includes orally administering venetoclax at a dose of 100 mg on day 1 of the cycle, 200 mg on day 2 of the cycle, and 400 mg on days 3 through 28 of the cycle.

[0387] In one case of I129 or I130 (I132), the method further includes administering venetoclax at an oral daily dose of 400 mg.

[0388] In one of the cases from I125 to I132 (I133), the hematological malignancy was AML. In one of the cases from I125 to I132 (I134), the hematological malignancy was BPDCN. In one of the cases from I125 to I132 (I135), the hematological malignancy was chronic myelomonocytic leukemia (CMML). In one of the cases from I125 to I132 (I136), the hematological malignancy was myelofibrosis (MF). In one of the cases from I125 to I132 (I137), the hematological malignancy was myelodysplastic syndrome (MDS). * * *

[0389] It should be understood that the detailed description section, rather than the summary and abstract section, is intended to be used to interpret the claims. The summary and abstract section presents one, but not all, exemplary embodiments of the invention, as intended by the inventors, and is therefore not intended to limit the invention and the appended claims in any way.

[0390] The present invention has been described above using functional components that demonstrate the implementation and relationships of specified functions. The boundaries of these functional components are arbitrarily defined herein for the sake of clarity. Alternative boundaries can be defined as long as the specified functions and their relationships are adequately performed.

[0391] The foregoing description of specific embodiments will fully reveal the general nature of the invention, and others may readily modify and / or adapt such specific embodiments for various uses without departing from the general concept of the invention and without unnecessary experimentation, by applying knowledge in the art. Accordingly, such adaptations and modifications are intended to be within the meaning and scope of equivalents of the disclosed embodiments based on the teachings and guidance presented herein. It should be understood that the terms and phrases herein are for illustrative purposes only, and not for limitation, so that they may be interpreted by those skilled in the art in light of the teachings and guidance.

[0392] The scope and breadth of the present invention should not be limited by any of the exemplary embodiments described above, but should be defined solely in accordance with the following claims and their equivalents. In certain embodiments, for example, the following items are provided: (Item 1) A method for treating hematological malignancies in a subject, comprising administering to a subject in need of such treatment an immune complex that binds to CD123 (the immune complex includes an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region CDR1 containing the amino acid sequence of SEQ ID NO: 5, a heavy chain variable region CDR2 containing the amino acid sequence of SEQ ID NO: 6, a heavy chain variable region CDR3 containing the amino acid sequence of SEQ ID NO: 7, a light chain variable region CDR1 containing the amino acid sequence of SEQ ID NO: 8, a light chain variable region CDR2 containing the amino acid sequence of SEQ ID NO: 9, and a light chain variable region CDR3 containing the amino acid sequence of SEQ ID NO: 10), a BCL-2 inhibitor, a hypomethylating agent, or a combination thereof. (Item 2) The method according to item 1, wherein the immune complex is administered in combination with the BCL-2 inhibitor. (Item 3) The method according to item 1, wherein the immune complex is administered in combination with the hypomethylating agent. (Item 4) The method according to item 1, wherein the immune complex is administered in combination with the BCL-2 inhibitor and the hypomethylating agent. (Item 5) The method according to any one of items 1 to 4, wherein the antibody or its antigen-binding fragment comprises VH containing the amino acid sequence shown in SEQ ID NO: 1 and / or VL containing the amino acid sequence shown in SEQ ID NO: 2. (Item 6) The method according to any one of items 1 to 4, wherein the antibody or antigen-binding fragment comprises a heavy chain containing the amino acid sequence shown in SEQ ID NO: 3 and / or a light chain containing the amino acid sequence shown in SEQ ID NO: 4. (Item 7) The method according to any one of items 1 to 6, wherein the immune complex comprises a cytotoxin, optionally the cytotoxin is a DNA alkylating agent, and optionally the DNA alkylating agent is an indolino-benzodiazepine (IGN) DNA alkylating agent. (Item 8) The method according to any one of items 1 to 7, wherein the immune complex is IMGN632. (Item 9) The aforementioned immune complex is administered in a pharmaceutical composition containing an immune complex having the following structure: [ka] , in the formula, The method according to any one of items 1 to 7, wherein G4723A comprises a heavy chain containing the amino acid sequence shown in SEQ ID NO: 3 and a light chain containing the amino acid sequence shown in SEQ ID NO: 4. (Item 10) The method according to any one of items 1 to 9, wherein the administration is a frontline therapy. (Item 11) The method according to any one of items 1 to 10, wherein the immune complex is administered intravenously. (Item 12) The method according to any one of items 1 to 11, wherein administration of the immune complex with the BCL-2 inhibitor, the hypomethylating agent, or a combination thereof produces a synergistic effect. (Item 13) The method according to any one of items 1, 2, and 4-12, wherein the administration of the immune complex and the BCL-2 inhibitor does not produce more toxicity than the administration of the immune complex alone or the BCL-2 inhibitor alone. (Item 14) The method according to any one of items 1 and 3 to 12, wherein the administration of the immune complex and the hypomethylating agent does not produce more toxicity than the administration of the immune complex alone or the hypomethylating agent alone. (Item 15) The method according to any one of items 1 and 4-12, wherein the administration of the immune complex, the BCL-2 inhibitor, and the hypomethylating agent does not produce more toxicity than the administration of the immune complex, the BCL-2 inhibitor, and / or the hypomethylating agent. (Item 16) The method according to any one of items 1, 2, and 5-15, wherein the immune complex is administered once in a 21-day cycle. (Item 17) The method according to item 16, wherein the immune complex is administered once in the 21-day cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg, and optionally, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg. (Item 18) The method according to item 17, wherein the immune complex is administered once in the 21-day cycle at a dose of approximately 0.045 mg / kg. (Item 19) The method according to item 17, wherein the immune complex is administered once in the 21-day cycle at a dose of approximately 0.03 mg / kg. (Item 20) The method according to item 17, wherein the immune complex is administered once in the 21-day cycle at a dose of approximately 0.015 mg / kg. (Item 21) The method according to any one of items 1, 2, and 4-20, wherein the BCL-2 inhibitor is administered daily in a 21-day cycle. (Item 22) The method according to any one of items 1, 2, and 4-20, wherein the BCL-2 inhibitor is administered in doses of 100 mg on day 1 of a 21-day cycle, 200 mg on day 2 of the 21-day cycle, and 400 mg on days 3 through 21 of the 21-day cycle. (Item 23) The method according to any one of items 1, 2, and 4-20, wherein the BCL-2 inhibitor is administered in a dose of 100 mg on day 1 of a 21-day cycle, 200 mg on day 2 of the 21-day cycle, and 400 mg on days 3-7 or 3-14 of the 21-day cycle. (Item 24) The method according to any one of items 1 and 3-15, wherein the immune complex is administered once in a 28-day cycle. (Item 25) The method according to item 24, wherein the immune complex is administered once in the 28-day cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg, and optionally, the dose is approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg. (Item 26) The method according to item 24, wherein the immune complex is administered once in the 28-day cycle at a dose of approximately 0.03 mg / kg. (Item 27) The method according to item 24, wherein the immune complex is administered once in the 28-day cycle at a dose of approximately 0.045 mg / kg. (Item 28) The method according to any one of items 1, 2, and 4-27, wherein the BCL-2 inhibitor is venetoclax. (Item 29) The method according to any one of items 1, 2, 4-21, and 23-28, wherein the BCL-2 inhibitor is administered in a dose of 400 mg. (Item 30) The method according to any one of items 1, 2, 4-21, and 23-28, wherein the BCL-2 inhibitor is administered in a dose of 200 mg. (Item 31) The method according to any one of items 23 to 30, wherein the BCL-2 inhibitor is administered daily in a 28-day cycle. (Item 32) The method according to any one of items 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 7 of the cycle. (Item 33) The method according to any one of items 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1-8 of the cycle. (Item 34) The method according to any one of items 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 14 of the cycle. (Item 35) The method according to any one of items 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 18 of the cycle. (Item 36) The method according to any one of items 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 21 of the cycle. (Item 37) The method according to item 31, wherein the BCL-2 inhibitor is administered daily on days 1 to 28 of the cycle. (Item 38) The method according to any one of items 24 to 28, wherein the BCL-2 inhibitor is administered in doses of 100 mg on day 1 of a 28-day cycle, 200 mg on day 2 of the 28-day cycle, and 400 mg on days 3 to 28 of the 28-day cycle. (Item 39) The method according to any one of items 24 to 28, wherein the BCL-2 inhibitor is administered in a dose of 100 mg on day 1 of a 28-day cycle, 200 mg on day 2 of the 28-day cycle, and 400 mg on days 3 to 7 or 3 to 14 of the 28-day cycle. (Item 40) The method according to any one of items 24 to 28, wherein the BCL-2 inhibitor is administered in a dose of 100 mg on day 1 of the 28-day cycle and in a dose of 200 mg on days 2 to 28, 2 to 14, or 2 to 7 of the 28-day cycle. (Item 41) The method according to any one of items 1, 2, and 4-40, wherein the BCL-2 inhibitor is administered orally. (Item 42) The method according to any one of items 1, 2, and 4-41, wherein the administration of the immune complex is initiated 6 days after the commencement of the administration of the BCL-2 inhibitor. (Item 43) The method according to any one of items 1, 3-15, and 23-42, wherein the low-methylating agent is azacitidine. (Item 44) The method according to item 43, wherein the azacitidine is administered in a 28-day cycle. (Item 45) The method according to item 43 or 44, wherein the azacitidine is administered once daily on days 1 to 7 of a 28-day cycle. (Item 46) The method according to item 43 or 44, wherein the azacitidine is administered once daily on days 1 to 5 of a 28-day cycle. (Item 47) The method according to item 43 or 44, wherein the azacitidine is administered once daily on days 3 through 7 of a 28-day cycle. (Item 48) The aforementioned azacitidine is approximately 75 mg / m² 2 The method described in any one of items 43 to 47, administered in the specified dose. (Item 49) The method according to any one of items 43 to 48, wherein the azacitidine is administered subcutaneously or intravenously. (Item 50) The method according to any one of items 1 and 3 to 42, wherein the low-methylating agent is decitabine. (Item 51) The method according to item 50, wherein the decitabine is administered intravenously. (Item 52) The method according to any one of items 1 to 51, wherein the aforementioned hematological malignancy is present in the subject as minimal residual disease (MRD). (Item 53) A method for treating hematological malignancies that exist as minimal residual disease in human subjects, comprising administering to the subject an anti-CD123 immune complex containing an anti-CD123 antibody or an antigen-binding fragment linked to a cytotoxic agent. (Item 54) The method according to item 53, wherein the immune complex is administered in a dose of approximately 0.045 mg / kg to approximately 0.18 mg / kg. (Item 55) The method according to item 53, wherein the immune complex is administered at a dose of approximately 0.045 mg / kg. (Item 56) The method according to item 53 or 54, wherein approximately 0.045 mg / kg, approximately 0.09 mg / kg, approximately 0.135 mg / kg, or approximately 0.18 mg / kg is administered to the subject. (Item 57) The method according to any one of items 53 to 56, wherein the immune complex is administered intravenously. (Item 58) The method according to any one of items 53 to 57, wherein the aforementioned hematological malignancy is leukemia. (Item 59) The method according to any one of items 53 to 58, wherein the immune complex is administered to the subject once in a 21-day cycle. (Item 60) The method according to any one of items 16-52 or 59, wherein the administration constitutes one cycle. (Item 61) The method according to any one of items 16-52 or 59, wherein the administration is two or more cycles, and optionally the administration is at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten cycles, or the administration is about two to four cycles, about two to six, about two to eight, about two to ten, or about two to twelve cycles. (Item 62) The method according to any one of items 1 to 61, further comprising administering a reduced dose of the immune complex after dose-limiting toxicity has occurred in the subject and been reduced to baseline or grade 2 or less. (Item 63) The method according to any one of items 1 to 62, wherein the immune complex is further administered as maintenance therapy. (Item 64) The method according to item 63, wherein the maintenance therapy comprises administering the immune complex once every 21 days. (Item 65) The method according to item 64, wherein the maintenance therapy comprises administering the immune complex once in the 21-day cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg, and optionally, the dose being approximately 0.015 mg / kg, approximately 0.045 mg / kg, or approximately 0.09 mg / kg. (Item 66) The method according to item 64, wherein the maintenance therapy comprises administering the immune complex once in the 21-day cycle at a dose of approximately 0.045 mg / kg. (Item 67) The method according to any one of items 1-9, 11-50, and 60-66, wherein the hematological malignancy is a recurrent hematological malignancy. (Item 68) The method according to any one of items 1-9, 11-50, and 60-67, wherein the hematological malignancy is a refractory hematological malignancy. (Item 69) The method according to any one of items 1 to 68, wherein the hematological malignancy is acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia in acute transformation / acute transformation phase (BP-CML), and blastic plasmacytoid dendritic cell neoplasm (BPDCN). (Item 70) The method according to items 1 to 68, wherein the hematological malignancy is AML. (Item 71) The method according to any one of items 1 to 68, wherein the hematological malignancy is BPDCN. (Item 72) The method according to any one of items 1 to 68, wherein the hematological malignancy is ALL. (Item 73) The method according to any one of items 1 to 72, wherein the hematological malignancy is a hematological malignancy that expresses CD123. (Item 74) The method according to any one of items 1 to 73, wherein CD123 is detected in the sample obtained from the hematological malignancy before administration, and optionally, CD123 is detected by flow cytometry. (Item 75) The method according to any one of items 1 to 74, further comprising detecting CD123 in a sample obtained from the hematological malignancy before administration. (Item 76) The method according to any one of items 1 to 75, wherein at least 80% of the cells in the hematological malignancy express CD123. (Item 77) The method according to any one of items 1 to 76, wherein CD123 is detected in at least 80% of the cells in the sample obtained from the hematological malignancy before administration. (Item 78) The method according to any one of items 1 to 77, further comprising detecting CD123 in at least 80% of the cells in the sample obtained from the hematological malignancy before the administration. (Item 79) The method according to any one of items 1-52 and 60-78, wherein the hematological malignancy is resistant to IMGN632. (Item 80) The method according to any one of items 1 to 79, wherein the hematological malignancy exhibits multidrug resistance 1 (MDR1). (Item 81) The method according to any one of items 1 to 80, wherein the hematological malignancy expresses P-glycoprotein (P-gp). (Item 82) The method according to any one of items 1 to 81, wherein the subject has an absolute neutrophil count greater than 500 / μL. (Item 83) The method according to any one of items 1-9 and 11-82, wherein the subject has received at least one prior therapy, at least two prior therapies, or at least three prior therapies. (Item 84) The method described in any one of items 1-9 and 11-83, wherein the cancer has been previously treated with venetoclax. (Item 85) The method described in any one of items 1 to 83, wherein the aforementioned cancer has not been previously treated with venetoclax. (Item 86) The method described in any one of items 1-9 and 11-85, wherein the cancer has been previously treated with a hypomethylating agent. (Item 87) The method described in any one of items 1 to 85, wherein the cancer has not been previously treated with a hypomethylating agent. (Item 88) The method according to any one of items 1 to 87, wherein the subject has been pre-treated with a corticosteroid prior to the administration of the immune complex, and optionally the corticosteroid is diphenhydramine, acetaminophen, paracetamol, dexamethasone, or a combination thereof. (Item 89) The method according to any one of items 1 to 87, further comprising pretreatment of the subject with a corticosteroid prior to administration of the immune complex, wherein the corticosteroid is optionally diphenhydramine, acetaminophen, paracetamol, dexamethasone, or a combination thereof. (Item 90) The method according to any one of items 1 to 52 and 60 to 89, wherein the immune complex and the BCL-2 inhibitor, the hypomethylating agent, or a combination thereof are administered in a separate pharmaceutical composition. (Item 91) The method described in any one of items 1 to 90, wherein the subject is a human. (Item 92) A method for treating hematological malignancies in human subjects, comprising administering IMGN632 and venetoclax to subjects in need of such treatment in a 21-day cycle, wherein IMGN632 is administered intravenously at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg on the 7th day of the cycle, and venetoclax is administered orally at a daily dose of 400 mg. (Item 93) The method described in item 92, wherein IMGN632 is administered at a dose of approximately 0.045 mg / kg. (Item 94) The method described in item 92, wherein IMGN632 is administered at a dose of approximately 0.03 mg / kg. (Item 95) A method for treating hematological malignancies in human subjects, comprising administering IMGN632 and azacitidine to subjects in need in a 28-day cycle, wherein IMGN632 is administered intravenously on the 7th day of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg, and azacitidine is administered at a dose of 75 mg / m². 2 The treatment method, administered subcutaneously or intravenously in the specified dose. (Item 96) The method according to item 95, wherein the azacitidine is administered on days 1 to 7 of the cycle. (Item 97) The method according to item 95, wherein the azacitidine is administered on days 1 to 5 of the cycle. (Item 98) The method described in any one of items 95-97, wherein IMGN632 is administered at a dose of approximately 0.045 mg / kg. (Item 99) The method described in any one of items 95-97, wherein IMGN632 is administered at a dose of approximately 0.03 mg / kg. (Item 100) The method described in any one of items 95-97, wherein IMGN632 is administered at a dose of approximately 0.015 mg / kg. (Item 101) The method according to any one of items 95 to 100, further comprising administering venetoclax at an oral daily dose of 400 mg. (Item 102) The method according to any one of items 92-94 and 101, wherein venetoclax is administered on days 1-7 of the cycle. (Item 103) The method according to any one of items 92-94 and 101, wherein venetoclax is administered on days 1-8 of the cycle. (Item 104) The method according to any one of items 92-94 and 101, wherein venetoclax is administered on days 1-14 of the cycle. (Item 105) The method according to any one of items 92-94 and 101, wherein venetoclax is administered on days 1-18 of the cycle. (Item 106) The method according to any one of items 92-94 and 101, wherein venetoclax is administered on days 1 to 21 of the cycle. (Item 107) The method according to item 101, wherein venetoclax is administered on days 1 to 28 of the cycle. (Item 108) The method according to any one of items 92 to 107, wherein the hematological malignancy is AML, BPDCN, or ALL.

Claims

1. A method for treating hematological malignancies in a subject, comprising administering to a subject in need of such treatment an immune complex that binds to CD123 (the immune complex includes an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region CDR1 containing the amino acid sequence of SEQ ID NO: 5, a heavy chain variable region CDR2 containing the amino acid sequence of SEQ ID NO: 6, a heavy chain variable region CDR3 containing the amino acid sequence of SEQ ID NO: 7, a light chain variable region CDR1 containing the amino acid sequence of SEQ ID NO: 8, a light chain variable region CDR2 containing the amino acid sequence of SEQ ID NO: 9, and a light chain variable region CDR3 containing the amino acid sequence of SEQ ID NO: 10), a BCL-2 inhibitor, a hypomethylating agent, or a combination thereof.

2. The method according to claim 1, wherein the immune complex is administered in combination with the BCL-2 inhibitor.

3. The method according to claim 1, wherein the immune complex is administered in combination with the hypomethylating agent.

4. The method according to claim 1, wherein the immune complex is administered in combination with the BCL-2 inhibitor and the hypomethylating agent.

5. The method according to any one of claims 1 to 4, wherein the antibody or its antigen-binding fragment comprises VH having the amino acid sequence shown in SEQ ID NO: 1 and / or VL having the amino acid sequence shown in SEQ ID NO:

2.

6. The method according to any one of claims 1 to 4, wherein the antibody or antigen-binding fragment comprises a heavy chain containing the amino acid sequence shown in SEQ ID NO: 3 and / or a light chain containing the amino acid sequence shown in SEQ ID NO:

4.

7. The method according to any one of claims 1 to 6, wherein the immune complex comprises a cytotoxin, optionally the cytotoxin is a DNA alkylating agent, and optionally the DNA alkylating agent is an indolino-benzodiazepine (IGN) DNA alkylating agent.

8. The method according to any one of claims 1 to 7, wherein the immune complex is IMGN632.

9. The aforementioned immune complex is administered in a pharmaceutical composition containing an immune complex having the following structure: 【Chemistry 1】 , in the formula, The method according to any one of claims 1 to 7, wherein G4723A comprises a heavy chain having the amino acid sequence shown in SEQ ID NO: 3 and a light chain having the amino acid sequence shown in SEQ ID NO:

4.

10. The method according to any one of claims 1 to 9, wherein the administration is a frontline therapy.

11. The method according to any one of claims 1 to 10, wherein the immune complex is administered intravenously.

12. The method according to any one of claims 1 to 11, wherein administration of the immune complex with the BCL-2 inhibitor, the hypomethylating agent, or a combination thereof produces a synergistic effect.

13. The method according to any one of claims 1, 2, and 4 to 12, wherein the administration of the immune complex and the BCL-2 inhibitor does not produce more toxicity than the administration of the immune complex alone or the BCL-2 inhibitor alone.

14. The method according to any one of claims 1 and 3 to 12, wherein the administration of the immune complex and the hypomethylating agent does not produce more toxicity than the administration of the immune complex alone or the hypomethylating agent alone.

15. The method according to any one of claims 1 and 4 to 12, wherein the administration of the immune complex, the BCL-2 inhibitor, and the hypomethylating agent does not produce more toxicity than the administration of the immune complex, the BCL-2 inhibitor, and / or the hypomethylating agent.

16. The method according to any one of claims 1, 2, and 5 to 15, wherein the immune complex is administered once in a 21-day cycle.

17. The method according to claim 16, wherein the immune complex is administered once in the 21-day cycle at a dose of about 0.015 mg / kg to about 0.09 mg / kg, and optionally the dose is about 0.015 mg / kg, about 0.045 mg / kg, or about 0.09 mg / kg.

18. The method according to claim 17, wherein the immune complex is administered once in the 21-day cycle at a dose of approximately 0.045 mg / kg.

19. The method according to claim 17, wherein the immune complex is administered once in the 21-day cycle at a dose of approximately 0.03 mg / kg.

20. The method according to claim 17, wherein the immune complex is administered once in the 21-day cycle at a dose of approximately 0.015 mg / kg.

21. The method according to any one of claims 1, 2, and 4 to 20, wherein the BCL-2 inhibitor is administered daily in a 21-day cycle.

22. The method according to any one of claims 1, 2, and 4 to 20, wherein the BCL-2 inhibitor is administered in a dose of 100 mg on day 1 of a 21-day cycle, 200 mg on day 2 of the 21-day cycle, and 400 mg on days 3 to 21 of the 21-day cycle.

23. The method according to any one of claims 1, 2, and 4 to 20, wherein the BCL-2 inhibitor is administered in a dose of 100 mg on day 1 of a 21-day cycle, 200 mg on day 2 of the 21-day cycle, and 400 mg on days 3 to 7 or 3 to 14 of the 21-day cycle.

24. The method according to any one of claims 1 and 3 to 15, wherein the immune complex is administered once in a 28-day cycle.

25. The method according to claim 24, wherein the immune complex is administered once in the 28-day cycle at a dose of about 0.015 mg / kg to about 0.09 mg / kg, and optionally, the dose is about 0.015 mg / kg, about 0.045 mg / kg, or about 0.09 mg / kg.

26. The method according to claim 24, wherein the immune complex is administered once in the 28-day cycle at a dose of approximately 0.03 mg / kg.

27. The method according to claim 24, wherein the immune complex is administered once in the 28-day cycle at a dose of approximately 0.045 mg / kg.

28. The method according to any one of claims 1, 2, and 4 to 27, wherein the BCL-2 inhibitor is venetoclax.

29. The method according to any one of claims 1, 2, 4-21, and 23-28, wherein the BCL-2 inhibitor is administered in a dose of 400 mg.

30. The method according to any one of claims 1, 2, 4-21, and 23-28, wherein the BCL-2 inhibitor is administered in a dose of 200 mg.

31. The method according to any one of claims 23 to 30, wherein the BCL-2 inhibitor is administered daily in a 28-day cycle.

32. The method according to any one of claims 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 7 of the cycle.

33. The method according to any one of claims 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 8 of the cycle.

34. The method according to any one of claims 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 14 of the cycle.

35. The method according to any one of claims 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 18 of the cycle.

36. The method according to any one of claims 21 and 28-31, wherein the BCL-2 inhibitor is administered daily on days 1 to 21 of the cycle.

37. The method according to claim 31, wherein the BCL-2 inhibitor is administered daily on days 1 to 28 of the cycle.

38. The method according to any one of claims 24 to 28, wherein the BCL-2 inhibitor is administered in a dose of 100 mg on day 1 of a 28-day cycle, 200 mg on day 2 of the 28-day cycle, and 400 mg on days 3 to 28 of the 28-day cycle.

39. The method according to any one of claims 24 to 28, wherein the BCL-2 inhibitor is administered in a dose of 100 mg on day 1 of a 28-day cycle, 200 mg on day 2 of the 28-day cycle, and 400 mg on days 3 to 7 or 3 to 14 of the 28-day cycle.

40. The method according to any one of claims 24 to 28, wherein the BCL-2 inhibitor is administered in a dose of 100 mg on day 1 of the 28-day cycle, and in a dose of 200 mg on days 2 to 28, 2 to 14, or 2 to 7 of the 28-day cycle.

41. The method according to any one of claims 1, 2, and 4 to 40, wherein the BCL-2 inhibitor is administered orally.

42. The method according to any one of claims 1, 2, and 4 to 41, wherein the administration of the immune complex is initiated 6 days after the commencement of the administration of the BCL-2 inhibitor.

43. The method according to any one of claims 1, 3 to 15, and 23 to 42, wherein the low-methylating agent is azacitidine.

44. The method according to claim 43, wherein the azacitidine is administered in a 28-day cycle.

45. The method according to claim 43 or 44, wherein the azacitidine is administered once daily on days 1 to 7 of a 28-day cycle.

46. The method according to claim 43 or 44, wherein the azacitidine is administered once daily on days 1 to 5 of a 28-day cycle.

47. The method according to claim 43 or 44, wherein the azacitidine is administered once daily on days 3 to 7 of a 28-day cycle.

48. The aforementioned azacitidine is approximately 75 mg / m² ２ The method according to any one of claims 43 to 47, administered in the specified dose.

49. The method according to any one of claims 43 to 48, wherein the azacitidine is administered subcutaneously or intravenously.

50. The method according to any one of claims 1 and 3 to 42, wherein the low-methylating agent is decitabine.

51. The method according to claim 50, wherein the decitabine is administered intravenously.

52. The method according to any one of claims 1 to 51, wherein the hematological malignancy is present in the subject as minimal residual disease (MRD).

53. A method for treating hematological malignancies that exist as minimal residual disease in human subjects, comprising administering to the subject an anti-CD123 immune complex containing an anti-CD123 antibody or an antigen-binding fragment linked to a cytotoxic agent.

54. The method according to claim 53, wherein the immune complex is administered in a dose of approximately 0.045 mg / kg to approximately 0.18 mg / kg.

55. The method according to claim 53, wherein the immune complex is administered at a dose of approximately 0.045 mg / kg.

56. The method according to claim 53 or 54, wherein approximately 0.045 mg / kg, approximately 0.09 mg / kg, approximately 0.135 mg / kg, or approximately 0.18 mg / kg is administered to the subject.

57. The method according to any one of claims 53 to 56, wherein the immune complex is administered intravenously.

58. The method according to any one of claims 53 to 57, wherein the hematological malignancy is leukemia.

59. The method according to any one of claims 53 to 58, wherein the immune complex is administered to the subject once in a 21-day cycle.

60. The method according to any one of claims 16 to 52 or 59, wherein the administration is one cycle.

61. The method according to any one of claims 16 to 52 or 59, wherein the administration is two or more cycles, and optionally the administration is at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten cycles, or the administration is about two to four cycles, about two to six, about two to eight, about two to ten, or about two to twelve cycles.

62. The method according to any one of claims 1 to 61, further comprising administering a reduced dose of the immune complex after dose-limiting toxicity has occurred in the subject and been reduced to baseline or grade 2 or lower.

63. The method according to any one of claims 1 to 62, wherein the immune complex is further administered as maintenance therapy.

64. The method according to claim 63, wherein the maintenance therapy comprises administering the immune complex once every 21 days.

65. The method according to claim 64, wherein the maintenance therapy comprises administering the immune complex once in the 21-day cycle at a dose of about 0.015 mg / kg to about 0.09 mg / kg, the dose being optionally about 0.015 mg / kg, about 0.045 mg / kg, or about 0.09 mg / kg.

66. The method according to claim 64, wherein the maintenance therapy comprises administering the immune complex once in the 21-day cycle at a dose of approximately 0.045 mg / kg.

67. The method according to any one of claims 1 to 9, 11 to 50, and 60 to 66, wherein the hematological malignancy is a recurrent hematological malignancy.

68. The method according to any one of claims 1 to 9, 11 to 50, and 60 to 67, wherein the hematological malignancy is a refractory hematological malignancy.

69. The method according to any one of claims 1 to 68, wherein the hematological malignancy is acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia in the acute transformation / acute transformation phase (BP-CML), and blastic plasmacytoid dendritic cell neoplasm (BPDCN).

70. The method according to claims 1 to 68, wherein the hematological malignancy is AML.

71. The method according to any one of claims 1 to 68, wherein the hematological malignancy is BPDCN.

72. The method according to any one of claims 1 to 68, wherein the hematological malignancy is ALL.

73. The method according to any one of claims 1 to 72, wherein the hematological malignancy is a hematological malignancy that expresses CD123.

74. The method according to any one of claims 1 to 73, wherein CD123 is detected in a sample obtained from the hematological malignancy before administration, and optionally, CD123 is detected by flow cytometry.

75. The method according to any one of claims 1 to 74, further comprising detecting CD123 in a sample obtained from the hematological malignancy before administration.

76. The method according to any one of claims 1 to 75, wherein at least 80% of the cells in the hematological malignancy express CD123.

77. The method according to any one of claims 1 to 76, wherein CD123 is detected in at least 80% of the cells in the sample obtained from the hematological malignancy before administration.

78. The method according to any one of claims 1 to 77, further comprising detecting CD123 in at least 80% of the cells in the sample obtained from the hematological malignancy before administration.

79. The method according to any one of claims 1 to 52 and 60 to 78, wherein the hematological malignancy is resistant to IMGN632.

80. The method according to any one of claims 1 to 79, wherein the hematological malignancy exhibits multidrug resistance 1 (MDR1).

81. The method according to any one of claims 1 to 80, wherein the hematological malignancy expresses P-glycoprotein (P-gp).

82. The method according to any one of claims 1 to 81, wherein the subject has an absolute number of neutrophils greater than 500 / μL.

83. The method according to any one of claims 1 to 9 and 11 to 82, wherein the subject has received at least one prior therapy, at least two prior therapies, or at least three prior therapies.

84. The method according to any one of claims 1 to 9 and 11 to 83, wherein the cancer has been previously treated with venetoclax.

85. The method according to any one of claims 1 to 83, wherein the cancer has not been previously treated with venetoclax.

86. The method according to any one of claims 1 to 9 and 11 to 85, wherein the cancer has been previously treated with a hypomethylating agent.

87. The method according to any one of claims 1 to 85, wherein the cancer has not been previously treated with a hypomethylating agent.

88. The method according to any one of claims 1 to 87, wherein the subject has been pre-treated with a corticosteroid prior to the administration of the immune complex, and optionally the corticosteroid is diphenhydramine, acetaminophen, paracetamol, dexamethasone, or a combination thereof.

89. The method according to any one of claims 1 to 87, further comprising pre-treating the subject with a corticosteroid before administration of the immune complex, wherein the corticosteroid is optionally diphenhydramine, acetaminophen, paracetamol, dexamethasone, or a combination thereof.

90. The method according to any one of claims 1 to 52 and 60 to 89, wherein the immune complex and the BCL-2 inhibitor, the hypomethylating agent, or a combination thereof are administered in a separate pharmaceutical composition.

91. The method according to any one of claims 1 to 90, wherein the subject is a human.

92. A method for treating hematological malignancies in human subjects, comprising administering IMGN632 and venetoclax to a subject in need of such treatment in a 21-day cycle, wherein IMGN632 is administered intravenously at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg on the 7th day of the cycle, and venetoclax is administered orally at a daily dose of 400 mg.

93. The method according to claim 92, wherein IMGN632 is administered at a dose of approximately 0.045 mg / kg.

94. The method according to claim 92, wherein IMGN632 is administered at a dose of approximately 0.03 mg / kg.

95. A method for treating hematological malignancies in human subjects, comprising administering IMGN632 and azacitidine to subjects in need in a 28-day cycle, wherein IMGN632 is administered intravenously on the 7th day of the cycle at a dose of approximately 0.015 mg / kg to approximately 0.09 mg / kg, and azacitidine is administered at a dose of 75 mg / m². ２ The treatment method, administered subcutaneously or intravenously in the specified dose.

96. The method according to claim 95, wherein the azacitidine is administered on days 1 to 7 of the cycle.

97. The method according to claim 95, wherein the azacitidine is administered on days 1 to 5 of the cycle.

98. The method according to any one of claims 95 to 97, wherein IMGN632 is administered at a dose of approximately 0.045 mg / kg.

99. The method according to any one of claims 95 to 97, wherein IMGN632 is administered at a dose of approximately 0.03 mg / kg.

100. The method according to any one of claims 95 to 97, wherein IMGN632 is administered at a dose of approximately 0.015 mg / kg.

101. The method according to any one of claims 95 to 100, further comprising administering venetoclax at an oral daily dose of 400 mg.

102. The method according to any one of claims 92-94 and 101, wherein venetoclax is administered on days 1 to 7 of the cycle.

103. The method according to any one of claims 92-94 and 101, wherein venetoclax is administered on days 1 to 8 of the cycle.

104. The method according to any one of claims 92-94 and 101, wherein venetoclax is administered on days 1 to 14 of the cycle.

105. The method according to any one of claims 92-94 and 101, wherein venetoclax is administered on days 1 to 18 of the cycle.

106. The method according to any one of claims 92-94 and 101, wherein venetoclax is administered on days 1 to 21 of the cycle.

107. The method according to claim 101, wherein venetoclax is administered on days 1 to 28 of the cycle.

108. The method according to any one of claims 92 to 107, wherein the hematological malignancy is AML, BPDCN, or ALL.

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