CD70 and venetoclax, BCL-2 inhibitor, combination therapy for treating acute myeloid leukemia

Abstract

To provide pharmaceutical compositions for use in combination therapies, particularly combination therapies for the treatment of myeloid malignancy.SOLUTION: The present invention provides a pharmaceutical composition for administering an antibody binding to CD70 or an antigen-binding fragment thereof in combination with a BCL-2 inhibitor, wherein the pharmaceutical composition includes an antibody binding to CD70 or an antigen-binding fragment thereof, the BCL-2 inhibitor is venetoclax or a pharmaceutically acceptable salt thereof, and the antibody binding to CD70 or an antigen-binding fragment thereof comprises a variable heavy chain (VH) domain and a variable light chain (VL) domain.SELECTED DRAWING: None

Description

[Technical field]

[0001] FIELD OF THEINVENTION The present invention relates to combination therapy, in particular for the treatment of myeloid malignancies. The combination therapy is particularly useful for the treatment of acute myeloid leukemia (AML). and a BCL-2 inhibitor, such as venetoclax or The combination therapy may further comprise an additional anti-cancer agent, such as an azathioprine. It may also include drugs used in the treatment of AML, such as cytidine or decitabine. [Background technology]

[0002] BACKGROUND OF THEINVENTION In recent years, the development of new cancer treatments has focused on molecular targets, especially proteins, involved in cancer progression. The list of molecular targets involved in tumor growth, invasion, and metastasis continues to expand. The list includes proteins overexpressed by tumor cells, as well as proteins involved in the vasculature and These molecular targets include targets related to systems that support tumor growth, such as the immune system. The number of therapeutic or anti-cancer drugs that are designed to interact with each other also continues to grow. Many targeted cancer drugs have been approved for clinical use, with many more in development. It's inside.

[0003] Due to its constitutive expression in many types of hematological malignancies and solid tumors, CD70 is It has been identified as a molecular target of particular interest (Junker et al. (2005) J Urol. 173:2150 -3; Sloan et al. (2004) Am J Pathol. 164:315-23; Held-Feindt and Mentlein (2002) Int J Cancer,98:352-6; Hishima et al. (2000) Am J Surg Pathol. 24:742 -6; Lens et al. (1999) Br J Haematol. 106:491-503; Boursalian et al. (2009) Adv Exp Med Biol. 647:108-119; Wajant H. (2016) Expert Opin Ther Targets, 20(8):959-973). CD70 is a type II transmembrane kinase that belongs to the tumor necrosis factor (TNF) superfamily. It is a type 1 glycoprotein that mediates its effects through binding to its cognate cell surface receptor, CD27. Both CD70 and CD27 are expressed by multiple cell types of the immune system, and CD70-CD2 7 signaling pathways have been implicated in the control of several different aspects of the immune response. Overexpression of CD70 is associated with a variety of autoimmune diseases, including rheumatoid and psoriatic arthritis, and lupus. This reflects the fact that it occurs in epidemic diseases (Boursalian et al., (2009) Adv Ex p Med Biol. 647:108-119; Han et al. (2005) Lupus,14(8):598-606; Lee et al. ( 2007) J Immunol. 179(4):2609-2615; Oelke et al. (2004) Arthritis Rheum. 50(6): 1850-1860).

[0004] CD70 expression is associated with poor prognosis in several cancers, including B-cell lymphoma, renal cell carcinoma, and breast cancer. It is said that this is related to the chromosome-specific markers (Bertrand et al., (2013) Genes Chromosomes Cancer, 52(8):764- 774;Jilaveanu et al. (2012) Hum Pathol. 43(9):1394-1399;Petrau et al. (2014) J Cancer, 5(9):761-764). CD70 expression was also found on metastatic tissue in a high percentage of cases. This indicates an important role of this molecule in cancer progression (Jacobs et al., 2015) Oncotarget,6(15):13462-13475). CD70 and its receptor CD2 on hematopoietic tumor cells Constitutive expression of CD70-CD27 directly regulates tumor cell proliferation and survival. It is believed to play a central role (Goto et al., (2012) Leuk Lymphoma, 53(8):1494-1 500; Lens et al. (1999) Br J Haematol. 106(2):491-503; Nilsson et al. (2005) Exp Hematol. 33(12):1500-1507; van Doorn et al. (2004) Cancer Res. 64(16):557 8-5586).

[0005] Upregulated CD70 expression on tumors, especially solid tumors that do not co-express CD27, has also been reported. , contribute to immune suppression in the tumor microenvironment in a variety of ways. For example, CD2 on regulatory T cells CD70 binding to CD7 increases Treg frequency and reduces tumor-specific T cell responses in mice. It has been shown that IL-15 reduces IL-15 expression and promotes tumor growth (Claus et al., (2012) Cancer Research 2013). s. 72(14):3664-3676). Thus, CD70-CD27 signaling also mediates immune responses by tumor-induced apoptosis of T lymphocytes. It can also attenuate the immune response (Chahlavi et al., (2005) Cancer Res. 65(12):5428-5438). Diegmann et al. (2006) Neoplasia, 8(11):933-938; Wischusen et al. (2002) Canc Finally, CD70 expression is also associated with T cell exhaustion, thereby Lymphocytes adopt a more differentiated phenotype and are unable to kill tumor cells. (Wang et al., (2012) Cancer Res, 72(23):6119-6129; Yang et al., (2014) Leukemia , 28(9):1872-1884).

[0006] Given the importance of CD70 in cancer progression, it is an attractive target for anticancer therapy. Antibodies targeting this cell surface protein are currently in clinical trials (Jacob et al., 2003). 015) Pharmacol Ther. 155:1-10; Silence et al. (2014) mAbs, 6(2):523-532). Summary of the Invention

[0007] (Summary of the invention) The present invention relates to combination therapies comprising antibodies or antigen-binding fragments thereof that bind to CD70. As shown in Figure 1, the list of proteins involved in tumor growth continues to expand, with more than one Combination therapies targeting these proteins are becoming increasingly attractive anticancer treatments. In the combination therapy of the present invention, the antibody or antigen-binding fragment thereof that binds to CD70 is 2 inhibitor, such as venetoclax or a pharma- ceutical acceptable salt thereof. Overexpression of BCL-2 in cancer cells confers resistance to apoptosis, Inhibition of this protein can promote tumor cell death. This compound is an example of a potent and selective small molecule inhibitor of the BCL-2 protein. As described above, a CD70-binding antibody or antigen-binding fragment thereof and a BCL-2 inhibitor, e.g. The combination with venetoclax or a pharma- ceutical acceptable salt thereof is useful in treating cancer, particularly acute myeloid leukemia. The present invention provides an effective therapy for the treatment of myeloid malignancies such as myeloid leukemia (AML).

[0008] In a first aspect, the present invention provides a method for the treatment of a cancer, comprising: (i) an antibody or antigen-binding fragment thereof that binds to CD70; and (ii) Combinations comprising a BCL-2 inhibitor are provided. In some preferred embodiments, The 2 inhibitor is compound (I) shown below: [ka] or a pharma- ceutical acceptable salt thereof. Compound (I) is also referred to herein as venetoclax.

[0009] In some embodiments, the antibody or antigen-binding fragment that binds to CD70 comprises: (i) a variable heavy chain A variable light chain (VH) domain and a variable light chain (VL) domain, the VH domain and the VL domain comprising: Heavy chain CDRs (HCDR3, HCDR2 and HCDR1) and light chain CDRs (LCDR3, LCDR2 and LCDR1): SEQ ID NO:3 HCDR3 comprising or consisting of SEQ ID NO:2; HCDR2 comprising or consisting of SEQ ID NO:2; HCDR1 comprising or consisting of SEQ ID NO:1; L comprising or consisting of SEQ ID NO:7 CDR3; LCDR2 comprising or consisting of SEQ ID NO:6; and (ii) an antibody or antigen-binding fragment comprising an LCDR1 consisting of SEQ ID NO: 4 and at least 7 VH domains that contain amino acid sequences that are 0%, at least 80%, at least 90%, or at least 95% identical Main, and at least 70%, at least 80%, at least 90%, at least an antibody or antigen-binding fragment comprising a VL domain comprising an amino acid sequence that is 95% identical; or ) ARGX-110. In some embodiments, the antibody is selected from IgG, preferably IgG The answer is 1.

[0010] The CD70 antibodies or antigen-binding fragments of the combination may also have one or more effector functions. In some embodiments, the antibody or antigen-binding fragment can be used to inhibit ADCC (antibody-dependent cell-mediated and / or comprises a defucosylated antibody domain; and / or or CDC (complement-dependent cytotoxicity) activity; and / or ADCP (antibody-dependent cellular phagocytosis) activity. In a preferred embodiment, the CD70 antibody is ARGX-110.

[0011] In some embodiments, the CD70 antigen-binding fragment of the combination comprises: an antibody light chain variable domain (VL); antibody heavy chain variable domain (VH); single chain antibody (scFv); F(ab')2 fragment; Fab fragment; Fd fragment; F v fragment; one-arm (monovalent) antibody; diabody, triabody, tetrabody, or Such antigen-binding fragments may be combined, assembled, or conjugated to form a and any antigen-binding molecule thereof.

[0012] In some embodiments, the CD70 antibody or antigen-binding fragment thereof and the BCL-2 inhibitor are In some embodiments, the CD70 antibody or its antigen-binding domain is formulated as a separate composition. The combination fragment and venetoclax or a pharma- ceutical acceptable salt thereof may be administered as separate compositions. It is formulated as follows.

[0013] The combinations of the present invention may also include one or more additional therapeutic agents, such as at least one additional anti-cancer agent, preferably In some embodiments, the additional anti-cancer agent may include a therapeutic agent for a myeloid malignancy. is a therapeutic agent for acute myeloid leukemia (AML). Methylating agents include, preferably, azacitidine or decitabine.

[0014] In a further aspect, the present invention provides a composition according to the first aspect of the invention for use in therapy. In particular, the present invention provides a combination for the treatment of malignancies, preferably myeloid malignancies, in human subjects. There is provided a combination according to the first aspect of the invention for use in therapy. A method for treating a malignancy, preferably a myeloid malignancy, in a human subject, comprising administering to the subject a compound according to the present invention. The present invention provides a method comprising administering an effective amount of any combination according to the first aspect of the present invention. .

[0015] The present invention also provides a method for the treatment of a malignancy, preferably a myeloid malignancy, in a human subject. and a BCL-2 inhibitor, preferably a compound of the above. The present invention provides an antibody molecule administered in combination with compound (I) or a pharma- ceutically acceptable salt thereof. The present invention also provides a BCL-2 inhibitor, preferably a medicament for use in the treatment of a malignant tumor in a human subject. and a compound (I) or a pharma- ceutical acceptable salt thereof, Compound (I) or a pharma- ceutical acceptable salt thereof is an antibody that binds to CD70 or an antibody thereof. The antibody is administered in combination with an original binding fragment.

[0016] The combinations of the present invention are particularly advantageous because they exhibit synergistic effects. In all aspects of the present invention, a CD70 antibody or an antigen-binding fragment thereof is preferably used in the present invention. and the dose at which a BCL-2 inhibitor is administered and / or provided in the combination. The respective doses used are selected so that the combination provides synergistic therapy.

[0017] In some preferred embodiments of the combinations of the invention, the CD70 antibody or antigen-binding fragment thereof The antibody and BCL-2 inhibitors were cultured in selected AML cell lines: NOMO-1, MOLM-13, NB4, and MV4-11. each present in the combination in an amount sufficient to provide synergistic cell killing when combined do.

[0018] With respect to the malignancies to be treated using the combinations of the present invention, the malignancies may be newly diagnosed relapsed or refractory myeloid malignant tumor; or acute myeloid leukemia ( AML); myelodysplastic syndromes (MDS); myeloproliferative neoplasms (MPN); chronic myeloid leukemia (CML); and chronic The malignant myeloid tumor may be selected from myelomonocytic leukemia (CMML). In an embodiment, the combination of the present invention is for the treatment of acute myeloid leukemia (AML).

[0019] In some embodiments, the subject or patient treated according to the methods of the present invention is The subjects were newly diagnosed AML patients who were ineligible for standard intensive chemotherapy. Patients aged 75 years or older who have been diagnosed with AML or who are newly diagnosed with AML are excluded from the use of standard intensive chemotherapy. The patient may also be an AML patient with comorbidities that may cause

[0020] In some embodiments, the CD70 antibody or antigen-binding fragment thereof is administered at a dose in the range of 0.1 to 25 mg / kg. Alternatively or in addition, BCL-2 The inhibitor, preferably venetoclax or a pharma- ceutical acceptable salt thereof, is administered in an amount of 100 mg to 600 mg In a preferred embodiment, the methods described herein are and administering a combination additionally comprising azacitidine, the azacitidine being 75 mg or less. g / m 2 In a further preferred embodiment, the methods described herein are administered in a dose of and administering a combination additionally comprising decitabine at a dose of 20 mg / m 2 For It is administered in doses.

[0021] In some embodiments, the method further comprises monitoring the patient's blast count. The patient's peripheral blood and / or bone marrow blast count can be reduced, e.g. For example, a reduction of 25% or less, for example, a reduction of 5% or less, for example, a reduction of 5% or less, for example, a reduction of minimal residual disease In some embodiments, the concentration of β-glucose in the blood may be reduced to a level below 100 mM, e.g., to an undetectable level. In the 12-month study, the bone marrow blast count decreased by between 5% and 25%, and the bone marrow blast count decreased by 1.5% compared to pretreatment. The sphere percentage is reduced by more than 50%.

[0022] In some embodiments, the method induces a partial response. In this method, the method induces a complete response, optionally accompanied by platelet and / or neutrophil recovery. The law requires that transfusions of red blood cells or platelets, or both, be continued for periods of 8 weeks or more, 10 weeks or more, or 12 weeks or more. In some embodiments, the method includes administering the drug to a patient after 30 days or after ... Reduces 60-day mortality.

[0023] In some embodiments, the method extends survival time. For example, the method comprises administering to the subject The combination of one or more therapeutic agents used in the treatment of certain myeloid malignancies is This method induces a negative minimal residual disease state. This may also be the case.

[0024] In some embodiments, the method further comprises providing a bone marrow transplant to the subject. Alternatively or additionally, the method may include administering one or more additional anti-cancer agents. The one or more additional cancer agents may further comprise a therapeutic agent for treating myeloid malignancies, preferably AML. The agent may be selected from any agent suitable for treatment. Preferred agents are selectin inhibitors. (e.g., GMI-1271); FMS-like tyrosine kinase receptor 3 (FLT3) inhibitors (e.g., Midstau cyclin-dependent kinase inhibitors; aminopeptidase inhibitors; JAK / STAT inhibitors; Cytarabine; anthracycline compounds (e.g., daunorubicin, idarubicin); doxorubicin Sorubicin; Hydroxyurea; Vyxeos; IDH1 or IDH2 inhibitors, Idohyfa (Idhifa) (or Enasidenib) or Tibsovo (or Ivosidenib), etc.; othened inhibitors, glasdegib, etc., BET bromodomain inhibitors, CD123 or CD33 targeting agents, H DAC inhibitor, LSC targeting agent, AML bone marrow niche targeting agent, and NEDD8 activating enzyme inhibitor, Pevone Dystat, etc. [Brief description of the drawings]

[0025] (drawing) [Figure 1] Co-treatment of cusatuzumab and decitabine synergistically eliminates NOMO-1 AML cells. NOMO-1 AML cells were treated with vehicle, cusatuzumab, venetoclax, or decitabine, alone or in fixed ratio combinations, in the presence of carboxyfluorescein succinimidyl ester (CFSE)-labeled NK cells (1:1 ratio). The number of NOMO-1 AML cells per well was counted after 72 hours, the degree of viability was determined by Annexin V staining, and the effect of drug treatment was calculated as the percentage of viable cells relative to vehicle-treated cells. Combination index (CI) values ​​were calculated and plotted against fraction affected (Fa) values ​​between 0 and 10. Fa-CI plots (Chou-Talalay plots) to assess synergy and / or antagonism are shown. Fa values ​​of 0, 0.5, and 1 correspond to 0%, 50%, and 100% dead cells. CI less than 1, CI being 1, and CI greater than 1 represent synergy, additivity, and antagonism, respectively. IC50 represents the Fa value at which the Ve / Cusa combination (low Fa) and Ve / De / Cusa combination (high Fa) reach the IC50 concentration. Ve / De, venetoclax and decitabine; De / Cusa, decitabine and cusatuzumab; Ve / Cusa, venetoclax and cusatuzumab; Ve / De / Cusa, venetoclax and decitabine and cusatuzumab.

[0026] [Diagram 2]Co-treatment of venetoclax and decitabine synergistically eliminates NOMO-1 AML cells. Individual CI-Fa plots of data for each combination are shown in Figure 1. (A) venetoclax and decitabine; (B) decitabine and cusatuzumab; (C) venetoclax and cusatuzumab; (D) venetoclax, decitabine and cusatuzumab.

[0027] [Diagram 3] Co-treatment of cusatuzumab and decitabine synergistically eliminates NB4 AML cells. NB4 AML cells were treated with vehicle, cusatuzumab, venetoclax, or decitabine, alone or in fixed ratio combinations, in the presence of CFSE-labeled NK cells (1:1 ratio). The number of NB4 AML cells per well was counted after 72 hours, the degree of viable cells was determined by Annexin V staining, and the effect of drug treatment was calculated as the percentage of viable cells relative to vehicle-treated cells. Combination index (CI) values ​​were calculated and plotted against effect ratio (Fa) values ​​between 0 and 10. Fa-CI plots [Chou-Talalay plots] to assess synergy and / or antagonism are shown. Fa values ​​of 0, 0.5, and 1 correspond to 0%, 50%, and 100% dead cells. (A) venetoclax and decitabine; (B) venetoclax and cusatuzumab; (C) decitabine and cusatuzumab; (D) venetoclax, decitabine and cusatuzumab.

[0028] [Figure 4]Co-treatment of cusatuzumab and decitabine synergistically eliminates MOLM-13 AML cells. MOLM-13 AML cells were treated with vehicle, cusatuzumab, venetoclax, or decitabine, alone or in fixed ratio combinations, in the presence of CFSE-labeled NK cells (1:1 ratio). The number of MOLM-13 AML cells per well was counted after 72 hours, the degree of viable cells was determined by Annexin V staining, and the effect of drug treatment was calculated as the percentage of viable cells relative to vehicle-treated cells. Combination index (CI) values ​​were calculated and values ​​were plotted against effect ratio (Fa) values. Fa-CI plots [Chou-Talalay plots] are shown to evaluate synergy and / or antagonism. Fa values ​​of 0, 0.5, and 1 correspond to 0%, 50%, and 100% dead cells. (A) venetoclax and decitabine; (B) venetoclax and cusatuzumab; (C) decitabine and cusatuzumab; (D) venetoclax, decitabine and cusatuzumab.

[0029] [Diagram 5] Co-treatment of cusatuzumab and decitabine synergistically eliminates MV4-11 AML cells. MV4-11 AML cells were treated with vehicle, cusatuzumab, venetoclax, or decitabine, alone or in fixed ratio combinations, in the presence of CFSE-labeled NK cells (1:1 ratio). The number of MV4-11 AML cells per well was counted after 72 hours, the degree of viable cells was determined by Annexin V staining, and the effect of drug treatment was calculated as the percentage of viable cells relative to vehicle-treated cells. Combination index (CI) values ​​were calculated and values ​​were plotted against effect ratio (Fa) values. Fa-CI plots [Chou-Talalay plots] are shown to evaluate synergy and / or antagonism. Fa values ​​of 0, 0.5, and 1 correspond to 0%, 50%, and 100% dead cells.

[0030] [Figure 6]Combined venetoclax and cusatuzumab treatment synergistically eliminates leukemic stem cells (LSCs) in vitro. CD34+CD38- AML LSCs from patients P1-P3 were cultured overnight in duplicate with cusatuzumab (Cusa: 0.3 μg / ml) or venetoclax (Ve: 6 nM), alone or in combination, in the presence of NK cells (1:1 ratio), and subsequently plated in methylcellulose. Colony formation was assessed after 14 days. (A) Absolute number of colonies per well from the first (1st) plating after treatment; (B) Cells harvested from the 1st plating were re-plated (2nd plating) and colony formation and colonies per well were assessed after 14 days. Data are shown as mean ± SD. Statistics: One-way ANOVA; Tukey's post-hoc test; *, P<0.05; **, P<0.01; ***, P<0.001.

[0031] [Figure 7] Combined venetoclax and cusatuzumab treatment synergistically eliminates LSCs in vitro. Data shown correspond to those in Figure 6 and are normalized by the mean number of colonies per well following vehicle treatment for each patient. (A) First plating; (B) Second plating.

[0032] [Figure 8] Combined venetoclax and cusatuzumab treatment synergistically eliminates LSCs in vitro. CD34+CD38- AML LSCs from patients P4 and P5 were cultured overnight in duplicate with cusatuzumab (Cusa: 0.3 μg / ml), venetoclax (Ve: 6 nM) or decitabine (0.01 μM), alone or in combination, in the presence of NK cells (1:1 ratio), and subsequently seeded in methylcellulose. Colony formation was assessed after 14 days. Data are shown as mean ± SD. (A) Results from single patients; (B) Results from patient P4 and patient P5.

[0033] [Figure 9]CD70 mRNA expression (as a percentage of housekeeping gene) after 24 or 48 h treatment with vehicle (Veh) or venetoclax (Ven). [Figure 10] CD70 protein expression and CD70 mRNA expression by MOLM-13 and NOMO-1 cells in the presence (grey bars) and absence (black bars) of venetoclax. Significance was determined using Student's t-test. MFI = mean fluorescence intensity. ***P<0.001. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] (Detailed Description) (A.Definition) Unless otherwise defined, all technical and scientific terms used herein are It has the same meaning as commonly understood by a person skilled in the art of the present invention.

[0035] "Combination Therapy" - As used herein, the term "combination therapy" refers to a combination of "Combination" refers to a treatment in which two or more therapeutic agents are administered to a human subject. For use in combination therapy. Two or more therapeutic agents are typically used to treat a single disease. The present invention relates to a method for treating a cancer or malignant tumor, comprising administering to said patient a therapeutically effective amount of the compound of the present invention. Or the combination therapy comprises an antibody or antigen-binding fragment that binds CD70 and a BCL-2 inhibitor, preferably or the small molecule inhibitor venetoclax or a pharma- ceutical acceptable salt thereof. As described above, the drugs included in the combination therapy may be administered to a subject or patient in need thereof. For administration, the compounds may be co-formulated for administration or may be provided separately, e.g., as separate compositions. may be provided.

[0036] "Antibody" - As used herein, the term "antibody" refers to full-length antibodies and variants thereof. The term "antibody" is intended to encompass modified, humanized, and germline antibodies. The term "antibody" as typically used herein includes, but is not limited to, "Antibody" refers to an immunoglobulin polypeptide having a combination of two heavy chains and two light chains. The polypeptide exhibits significant specific immune response to an antigen of interest (herein, CD70). As for the IgG class of antibodies, these antibodies have a molecular weight of about 23,000 daltons and a molecular weight of 23,000 daltons. It contains one identical light polypeptide chain and two identical heavy chains with molecular weights of 53,000 to 70,000. The four chains are joined by disulfide bonds in a "Y" configuration, where the light chain is " and brackets the heavy chain across the variable region. Antibody light chains are classified as either kappa or lambda (κ, λ). Each heavy chain class has In general, light and heavy chains may be associated with either kappa or lambda light chains. , covalently linked to each other, and the "tails" of these two heavy chains are held together by covalent disulfide bonds. or immunoglobulins derived from hybridomas, B cells or genetically engineered host cells In the heavy chain, the α-terminal end of the ribosome is bound to the α-terminal end of the ribosome by a non-covalent bond. The amino acid sequences run from N-terminus at the forked ends of the Y configuration to C-terminus at the bottom of each chain.

[0037] Those skilled in the art will appreciate that heavy chains are classified into gamma, mu, amyloid, α-gamma, β ... By understanding that the four chromosomes are classified as γ, μ, α, δ, or epsilon, Depending on the nature of the chains, the "classes" of antibodies are IgG, IgM, IgA, IgD, or IgM, respectively. E. Immunoglobulin subclass (isotype), e.g. IgG1, IgG2, IgG3 , IgG4, IgA1, etc. are well characterized and are known to have functional specialization. As used herein, the term "antibody" refers to an antibody from any class or subclass of antibody. It embraces the body.

[0038] "Antigen-binding fragment" - As used herein, the term "antigen-binding fragment" refers to an antigen-binding fragment of a full-length antibody. An antibody that contains a portion or part, or fewer amino acid residues than an intact or complete antibody. An antigen-binding fragment of an antibody is a fragment of the antibody that is an antibody but retains antigen-binding activity. and peptide fragments that exhibit specific immune reaction activity against the same antigen (eg, CD70). As used herein, the term "antigen-binding fragment" refers to an antibody light chain variable domain (VL); Single chain variable domain (VH); single chain antibody (scFv); F(ab')2 fragment; Fab fragment; Fd fragment; Fv fragment; single chain Arm (monovalent) antibodies; diabodies, triabodies, tetrabodies or the like Any antigen-binding fragment formed from a combination, assembly or conjugation of antigen-binding fragments. The term "antibody fragment" is intended to encompass an antibody fragment selected from the group consisting of: The term "antigen-binding fragment" as used herein also refers to unibodies, domain antibodies, and nanobodies. The fragment may be intended to encompass an antibody fragment selected from the group consisting of: By chemical or enzymatic treatment of intact or complete antibodies or antibody chains, or by recombinant techniques. It can be obtained by steps.

[0039] "Specific" and "Multispecific" Antibodies - for use in the combination therapy described herein The antibodies and antigen-binding fragments bind to a specific target antigen, such as CD70. Preferably, the antigens “specifically bind” to their target antigens, and the term “specifically binds” By "antibody" is meant any antibody or antigen binding molecule that preferentially immunoreacts with a given target, such as CD70. The antibodies and antigen-binding fragments of the present combinations and methods may be monospecific or The combination may contain one or more binding sites that specifically bind to a particular target. The antibodies and antigen-binding fragments of the present invention may be in a "multispecific antibody" format, e.g., a bispecific antibody. It is also possible to incorporate heterologous antibodies, where the multispecific antibodies are capable of binding to two or more target antigens. To achieve multispecificity, a "multispecific antibody" typically binds different VH - engineered to contain different combinations or pairings of heavy and light chain polypeptides having VL pairs Multispecific, and in particular bispecific, antibodies can be produced by combining the functions of native antibodies, e.g., Fc regions. The overall structure, such as a Y-shaped antibody with conjugated Fab arms of different specificities, is Alternatively, multispecific antibodies, e.g., bispecific antibodies, can be engineered to For example, multiple variable domains or variable domain pairs with different specificities are arranged on both ends of the Fc region. The molecule may be engineered to adopt a non-native conformation, such as that depicted in FIG.

[0040] "Modified Antibody" - As used herein, the term "modified antibody" refers to an antibody that is not naturally occurring. and synthetic forms of modified antibodies that do not contain at least two heavy chain portions, e.g., Antibodies that do not contain a complete heavy chain (such as domain deleted antibodies or minibodies); (two or more different antibodies) Multiple specificities of antibodies (modified to bind to different antigens or to different epitopes on a single antigen) isomeric forms (e.g., bispecific, trispecific, etc.); heavy chain molecules linked to scFv molecules, etc. scFv molecules are known in the art and are described, for example, in US Pat. No. 5,892,019. In addition, the term "modified antibody" refers to multivalent forms of antibodies (e.g., three or more copies of the same antigen). In another embodiment, the modified antibody of the present invention is an antibody that binds to a specific target polypeptide. The antibody comprises at least one heavy chain portion lacking a CH2 domain and is a ligand of a receptor pair. It is a fusion protein that includes a polypeptide binding domain that includes a binding portion of the member.

[0041] "Humanizing substitutions" - As used herein, the term "humanizing substitutions" refers to substitutions in the VH or VL domains of an antibody. The amino acid residues present at specific positions in the sequence correspond to the equivalent positions in a reference human VH or VL domain. This refers to an amino acid substitution in which the amino acid residue is replaced by an amino acid residue occurring in the reference human VH or VL domain. The main domain may be a human germline encoded VH or VL domain. The substitutions may be made in the framework regions and / or CDRs of the antibody, as defined herein. It is possible.

[0042] "Humanized variant" - As used herein, the term "humanized variant" or "human "Variant antibody" refers to a variant antibody that contains one or more "humanization substitutions" compared to a reference antibody; Here, a portion of the reference antibody (e.g., a VH domain and / or a VL domain or at least The CDRs (parts thereof including at least one CDR) have amino acids derived from a non-human species and are referred to as "humanizing substitutions." " occurs within an amino acid sequence derived from a non-human species.

[0043] "Germline-linked variant" - As used herein, the term "germline-linked variant" refers to a Specifically, a "germlined antibody" or "germlined antibody" refers to an antibody that has been modified by "humanization substitutions" therein. One or more amino acid residues present at a particular position(s) in the VH or VL domain of an antibody are The amino acid sequence of the nucleotide sequence that occurs at the equivalent position in a reference human VH or VL domain encoded by the human germline "Humanized variants" refer to any given "germline variant" in which the amino acid residues have been replaced with Regarding the "germline variant," the replacement amino acid residue that was replaced with the germline variant The group is composed exclusively or primarily of a single human germline-encoded VH or VL domain. The terms "humanized variant" and "germline variant" are used interchangeably. The terms "humanized" and "humanized" are often used interchangeably. , llama-derived) VH or VL domains, resulting in camelid (llama)-derived VH or VL A "humanized variant" of the domain is created. The amino acid residues substituted are Derived primarily or exclusively from a single human germline-encoded VH or VL domain sequence If so, the result is a "human germline cloning" of VH or VL domains derived from camelids (llamas). It will be "variant".

[0044] "CD70" - As used herein, the term "CD70" or "CD70 protein" or "CD The term "TNF ligand family 70 antigen" is used interchangeably and is a ligand for TNFRSF7 / CD27. CD70 refers to a member of the human CD70 protein. CD70 is also known as CD27L or TNFSF7. The terms "human CD70 protein" or "human CD70 antigen" or "human CD70" are used interchangeably and specifically refer to human Native human CD70 protein naturally expressed in vivo and / or on the surface of cultured human cell lines The term refers to human CD70 proteins, as well as human homologues, including recombinant forms and fragments thereof. Specific examples of human CD70 include: The amino acid sequence shown in NCBI reference sequence accession number NP_001243, or the extracellular domain thereof The present invention includes polypeptides which

[0045] "BCL-2 family" - As used herein, the term "BCL-2 family" or "BC The BCL-2 protein family is a set of pro-apoptotic and apoptotic proteins related to BCL-2. This refers to a collection of catalysis-inhibiting proteins, as described by Delbridge et al. (2016) See Rev Cancer. 16(2): 99-109. At least 16 members of this family However, there are three functional groups: (i) BCL-2-like proteins (e.g., BCL-2, BCL-X L / BCL2L1, BCLW BCL2L 2, MCL2, BFL1 / BCL2A1); (ii) BAX and BAK; and (iii) BH3-only proteins (e.g., BI M, PUMA, BAD, BMF, BID, NOXA, HRK, BIK). Milli inhibits apoptosis-inhibitory members of the family (e.g., BCL-2, BCL-X L ) and this It plays an essential role in regulating the qualitative apoptotic pathway and is usually a pro-apoptotic member antagonizing genes (e.g., BAX and BIM) e.g., gene translocation, amplification, overexpression, and mutation Deregulation of BCL-2 family members by IL-1 has been observed in many cancers. The downstream effect of is often resistance to apoptosis, thereby promoting cancer growth.

[0046] "BCL-2" - As used herein, "BCL-2" or "BCL-2 protein" refers to human The first member of the BCL-2 protein family identified in B cell lymphoma The cDNA encoding human BCL-2 was cloned in 1986, and the sequence of this protein was determined. In 1988, its important role in inhibiting apoptosis was revealed. BCL-2 has been shown to be upregulated in several types of cancer. Activated by the t(14;18) chromosomal translocation in follicular lymphoma. Amplification of the BCL-2 gene also occurs. It also treats leukemias (e.g., CLL), lymphomas (e.g., B-cell lymphoma), and some solid tumors (e.g., small cell Human BCL-2 has been reported in different cancers, including lung cancer. ) and are designated as NCBI reference sequences NP_000624.2 and NP_000648.2 It has an array.

[0047] "BCL-2 Inhibitor" - As used herein, a BCL-2 inhibitor specifically inhibits the activity of BCL-2. Any agent, compound or molecule capable of inhibiting the apoptosis-inhibiting activity of BCL-2, particularly It refers to an agent, compound or molecule capable of inhibiting the activity of Examples of BCL-2 inhibitors used in cancer include B-cell lymphoma homology 3 (BH3) mimetic compounds (Merino et al., (2018) Cancer Cell. 34(6): 879-891). Specific BCL-2 inhibitors are listed below. Non-limiting examples include venetoclax, ABT-737 (Oltersdorf, T et al., (2005) N ature 435: 677-681), navitoclax / ABT-263 (Tse, C. et al., (2008) Cancer Res. 6 8: 3421-3428), BM-1197 (Bai, L. et al., (2014) PLoS ONE 9: e99404), S44563 (Nemati ,F. et al., (2014) PLoS ONE 9: e80836), BCL2-32 (Adam,A. et al., (2014) Blood 1 24: 5304), AZD4320 (Hennessy, EJ et al., (2015) ACS Medicinal Chemistry annual meeting https: / / www.acsmedchem. org / ama / orig / abstracts / mediabstractf2015.pdf abs tr. 24), and S55746 (International Standard Randomized Controlled Trials Register Number (ISRCTN) http: / / www.isrctn. com / ISRCTN04804337(2016)). Further examples of BCL-2 inhibitors include those described in Ashkenazi, A et al. (2017) Nature Reviews Drug Discovery 16: 273-284, cited herein by the present inventors. It is incorporated in the specification.

[0048] "Venetoclax" - As used herein, the term "venetoclax" refers to It refers to a compound that has a chemical structure. [ka] This compound is referred to herein as "Compound (I)." Venetoclax It is a potent, selective and orally bioavailable inhibitor of the CL-2 protein. The formula is C 45 H 50 It is C1N7O7S and has a molecular weight of 868.44. Venetoclax is poorly soluble in water. Venetoclax is chemically known as 4-(4-{[2-(4-chlorophenyl)-4,4-dimethyl Cyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-({3-nitro-4-[(tetrahydro 1H-pyrrolo[2,3-b]pyridine Venetoclax can be expressed as (5-yloxy)benzamide. Another name for venetoclax is ABT-1. 99; Chemical Name 1257044-40-8; GDC-0199.

[0049] Venetoclax was approved by the U.S. Food and Drug Administration (FDA) in 2015 as a treatment for patients with rheumatoid arthritis who received at least one prior dose of the drug. Treatment of adult patients with chronic lymphocytic leukemia (CLL) or small lymphocytic leukemia (SLL) treated with the Venetoclax is also approved in the United States for use in adults 75 years and older or for use in patients with a strengthened induction Newly diagnosed acute myeloid leukemia in adults with comorbidities that preclude the use of chemotherapy In combination with azacitidine or decitabine or low-dose cytarabine to treat acute myeloid leukemia (AML) It is approved for use in

[0050] "Myeloid malignancies" - As used herein, the term "myeloid malignancies" refers to tumors that develop in the hematopoietic stem cell. Myeloid malignancies or diseases refer to any clonal disorder of myeloid or progenitor cells. These include chronic and acute conditions. Chronic conditions include myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPNs), and N) and chronic myelomonocytic leukemia (CMML), while acute manifestations include acute myeloid leukemia (AML).

[0051] "Acute myeloid malignancy" - as used herein, means "acute myeloid leukemia" or " "AML" refers to a hematopoietic neoplasm involving bone marrow cells. AML is a type of hematopoietic neoplasm that is characterized by a reduced differentiation capacity of bone marrow cells. AML is characterized by the clonal expansion of progenitor cells. Patients with AML develop an accumulation of blast cells in the bone marrow. As used herein, a "blast cell" or simply a "blast" is a cell that has been destroyed. Blast cells are typically also the terminal myeloid progenitor cells of AML patients. AML is usually diagnosed when patients have more than 20% of the This is the case when the above blast cells are shown.

[0052] "Standard Intensive Chemotherapy" - As used herein, "standard intensive chemotherapy" (Hitachi Medical University) In the specification, this is referred to as "enhanced induction therapy" or "induction therapy" (also called "7+3" induction therapy). This refers to chemotherapy with high-dose cytarabine for 7 days followed by an anthracycline (e.g. Standard intensive chemotherapy is characterized by the administration of daunorubicin or idarubicin. The method is intended to provide eligible, newly diagnosed AML patients with chemotherapy, typically after successful chemotherapy. Given to induce complete remission of AML with the intent of undergoing stem cell transplantation in patients As described herein, all newly diagnosed AML patients receive standard consolidation chemotherapy. This does not mean that the patient is eligible for treatment.

[0053] "Leukemic stem cell" - As used herein, "leukemic stem cell" or "LSC" refers to a leukemic stem cell that is LSCs are a subset of blast cells associated with immune deficiency. LSCs are blast cells with stem cell properties that can give rise to leukemic disease. It can self-regenerate by causing leukemia, or it can be similar to the original disease but self-regenerating. They can also partially differentiate into non-LSC-type conventional blast cells that cannot regenerate. (primary) A frequency ranging from 1 in 10,000 to 1 in 1 million as a percentage of AML blast cells (Pollyea and Jordan, (2017), Blood, 129:1627-1635, cited in this paper.) LSCs are CD34+, CD38-, and optionally CD45- and / or CD LSCs can also be characterized as cells that are CD45dim, SSClo, CD90+CD34 + cells.

[0054] "Anti-cancer Agent" - As used herein, an anti-cancer agent is an agent that inhibits cancer growth directly or indirectly. The term "antibody" refers to any agent capable of preventing, inhibiting or treating a disease, including, but not limited to, These include chemotherapeutic agents, immunotherapeutic agents, angiogenesis inhibitors, radionuclides, and the like, many examples of which are described in the art. It is well known to those skilled in the art.

[0055] (B. Combination Therapy with CD70 Antibody and BCL-2 Inhibitor) The present invention relates to a method for treating a pulmonary artery disease comprising administering to a patient a therapeutically effective amount of a therapeutic agent comprising: (i) an antibody or antigen-binding fragment thereof that binds to CD70; and (ii) a BCL-2 inhibitor. The present invention provides a combination therapy including As described herein, CD70 has been characterized as an attractive target for anti-cancer therapy. CD70 is constitutively expressed in many types of hematological malignancies and solid tumors. Expression of CD70 has been associated with poor prognosis in several cancers. Bodies are being developed, some of which have progressed to clinical trials.

[0056] Antibodies targeting CD70 have potential applications in the treatment of myeloid malignancies, particularly acute myeloid leukemia (AML). It has been shown that CD70 antibodies, A, and B are particularly effective in treating patients with AML. Results of a Phase I / II clinical trial investigating RGX-110, a schizophrenic drug, have demonstrated its potential in this condition, particularly in patients with standard in newly diagnosed patients classified as ineligible for intensive chemotherapy (see WO 2018 / 229303) The surprising effectiveness of this drug was revealed in the clinical trial. The CD70 antibody, when used in combination with azacytidine, inhibits leukemic stem cells (LSCs) in AML patients. Examination of LSCs isolated from patients in the study showed that they showed a strong association with myeloid cells. We showed evidence of increased asymmetric division of LSCs, indicative of differentiation. Collectively, these results suggest that CD7 0 antibody depletes the LSC pool in AML patients, thereby improving prognosis for remission , and furthermore, the risk of recurrence is reduced.

[0057] The present invention combines a CD70 antibody, or antigen-binding fragment thereof, with a BCL-2 inhibitor. The BCL-2 protein is a member of the BCL-2 family, which consists of more than 20 Members of the BCL-2 family are involved in the regulation of the intrinsic apoptosis pathway. It is involved in the regulation of the cellular cycle and plays a fundamental role in controlling the balance between cell survival and death.

[0058] BCL-2 proteins are apoptosis-inhibitory members of the BCL-2 family and are involved in the regulation of many different BCL-2 is upregulated in several types of cancer. Overexpression of BCL-2 is a signaling pathway that allows tumor cells to differentiate into By capturing pro-apoptotic proteins, it is possible to avoid apoptosis. BCL-2 is highly expressed in many hematological malignancies, including chronic lymphocytic leukemia (CLL), leukemia, and lymphoma. It is a major pro-survival protein in diseases such as follicular lymphoma and mantle cell lymphoma. Inhibiting BCL-2 may mediate anti-apoptotic or pro-survival effects of this protein. activity is also inhibited.

[0059] Inhibitory apoptosis members of the BCL-2 family, including BCL-2, are involved in the expression of BCL-2 in primary AML samples. It has been reported that it is overexpressed in the human leukemia (Bogenberger et al., (2014) Leukemia 28(2); 165 7-65) Overexpression of BCL-2 has also been reported in leukemic stem cells (LSCs) derived from AML patients. (Lagadinou et al., (2013) Cell Stem Cell 12(3); 329-341). Inhibition of BCL-2 led to selective eradication of quiescent LSCs (Lagadinou et al. (2013) Cell Stem Cell 12(3); 329-341).

[0060] Without being bound by theory, the combinations of the present invention may be used in combination with a CD70 antibody or antigen-binding fragment thereof. Due to the combined therapeutic effect of the combination of the serogroup and BCL-2 inhibitors, particularly at the level of LSCs, It is believed to be particularly effective for the treatment of AML. The self-renewal capacity of LSCs allows for the persistence of these cells. This means that it is a major factor contributing to disease recurrence.

[0061] As shown in the Examples, the combination of the present invention has synergistic therapeutic efficacy against AML cells, i.e., the level of inhibition induced by the combination is greater than the additive effect of the monotherapy alone; Methods for determining synergistic interactions are well known to those skilled in the art and are described in the Examples. The preferred method for determining whether a synergistic effect results from a combination is the Chou-Talal method. ay method (Chou, T. C., Cancer Res. (2011), incorporated by reference). 10) 70(2); 440-6).

[0062] The synergistic effect of the combination of the present invention is expressed as a strong inhibition of primary LSC cells from AML patients. Thus, the combination therapy of the present invention targets both the blast and LSC compartments. thereby improving the chances of disease remission while reducing the risk of relapse.

[0063] In some preferred embodiments of the combination of the invention, the BCL-2 inhibitor is venetoclave. Venetoclax is a small molecule inhibitor of BCL-2 or a pharma- ceutical acceptable salt thereof. and is described in US 2010 / 0305122, which is incorporated herein by reference. By inhibiting BCL-2, venetoclax inhibits the anti-apoptotic properties of this protein Venetoclax inhibits the survival-promoting activity of the majority of CLL cells and those that overexpress BCL-2. It rapidly induces apoptosis in lymphoma cell lines.

[0064] Early studies suggest venetoclax may be useful as an AML treatment (Konopleva et al., (2016) Cancer Discov. 6(10); 1106-17). LAX was found to have limited activity as a monotherapy. Subsequent studies Veneta combination with hypomethylating agents, i.e., azacitidine and decitabine. The effectiveness of these combinations has been investigated and these have been found to be particularly effective (Bogenbe (2015) Leuk Lymphoma, 56(1): 226-229. The clinical trials are planned to test combinations with either rituximab, decitabine, or low-dose cytarabine. The study was conducted (Dinardo et al., (2018) Lancet Oncol. 19(2): 216-228; Dinardo et al. (2019) Blood 133(1); 7-17). The results of these studies suggest that the Newly diagnosed acute myeloid leukemia in adults with comorbidities that preclude the use of intensive induction chemotherapy In combination with azacitidine, decitabine, or low-dose cytarabine for the treatment of leukemia (AML) This led to FDA approval of venetoclax for use in patients with rheumatoid arthritis.

[0065] In some other embodiments of the present invention, the BCL-2 inhibitor is B-cell lymphoma homology 3 (BH 3) Mimetic compounds. In some embodiments, the BCL-2 inhibitor is ABT-737, Navit is selected from clax, BM-1197, S44563, BCL2-32, AZD4320 or S55746.

[0066] (CD70 antibody) Antibodies or antigen-binding fragments that bind to CD70 and can be combined in any of the combinations described herein. Possible inclusions include, but are not limited to, antibodies that block the interaction of CD70 with CD27. a CD70 antibody or antigen-binding fragment that competes with CD27 for CD70 binding; Fragment; CD70 antibody or antigen-binding fragment that inhibits CD70-induced CD27 signaling; Treg activity CD70 antibodies or antigen-binding fragments that inhibit proliferation and / or proliferation; depleting CD70-expressing cells CD70 antibodies or antigen-binding fragments; CD70 antibodies or antigen-binding fragments that induce lysis of CD70-expressing cells; The present invention relates to a method for treating ADCC, CDC, and / or CD70-related diseases, comprising administering to a patient a therapeutically effective amount of a CD70 antibody or antigen-binding fragment that has ADCC, CDC functionality, and / or induces ADCP. nothing.

[0067] Exemplary CD70 antibodies are described in WO2012 / 123586, which is incorporated herein by reference. The previously reported ARGX-110 and SGN-70 (WO2006 / 113909 and McEarChern et al. (2008) Clin Cancer Res. 14(23):7763, both of which are incorporated herein by reference), and WO No. 6,200,254, and WO2006 / 044643 and WO2007 / 038637, each of which is incorporated herein by reference. These are the CD70 antibodies that were selected.

[0068] WO2006 / 044643 describes CD70 antibodies that contain an antibody effector domain, which , ADCC, ADCP, or CDC; and They exert growth-suppressing or cytotoxic effects, or act to protect against CD70-expressing immunological disorders. It can either exert an immunosuppressive effect or a cytostatic effect. The antibodies exemplified herein are 1F6 and 2F2, and do not need to be conjugated to a damaging agent. The antibodies are based on the antigen-binding regions of two monoclonal antibodies described herein.

[0069] WO2007 / 038637 describes a fully human monoclonal antibody that binds to CD70. The antibody was 1 × 10 -7 K below M D These antibodies are characterized by their binding to human CD70 at 100 ng / mL. The antibodies also bind to and thereby induce CD70-expressing renal cell carcinoma tumor cell lines, such as 786-O. It will be internationalized.

[0070] ARGX-110 is an IgG1 anti-CD70 antibody, also known as cusatuzumab. ARGX-110 inhibits CD70 It has been shown that IL-16 inhibits the interaction of IL-16 with its receptor CD27 (Silence et al., (2014) MAbs. Mar-Apr;6(2):523-32, incorporated herein by reference). In particular, ARGX-11 0 was shown to inhibit CD70-induced CD27 signaling. The levels are described, for example, in Riether et al. (J. Exp. Med. (2017) 214(2); 359-380). The CD27 antibody was detected by measuring serum soluble CD27 or by using the method described by Silence et al. (MAbs(2014) 6(2): 523-32). This can be determined by measuring IL-8 expression, as described above. However, inhibition of CD27 signaling reduced the activation and / or proliferation of Treg cells, which AGRX-110 also inhibits CD70 expression, which may reduce the inhibition of anti-tumor effector T cells. In particular, ARGX-110 was shown to inhibit antibody-dependent CD70-expressing tumor cells are targeted via ADCC and complement-dependent cytotoxicity (CDC). It has been shown to lyse CD70-expressing cells and also to increase antibody-dependent cellular phagocytosis (ADCP) of CD70-expressing cells. (Silence et al., op. cit.).

[0071] The CDR, VH and VL amino acid sequences of ARGX-110 or cusatuzumab are shown in the table below. (Table 1) [Table 1]

[0072] In some embodiments, the antibody or antigen-binding fragment thereof that binds to CD70 comprises a variable heavy chain A variable light chain domain (VH) and a variable light chain domain (VL), wherein the VH domain and the VL domain and the following CDR sequences: HCDR3 comprising or consisting of SEQ ID NO:3; HCDR2 comprising or consisting of SEQ ID NO:2; HCDR1 comprising or consisting of SEQ ID NO:1; LCDR3 comprising or consisting of SEQ ID NO:7; LCDR2 comprising or consisting of SEQ ID NO:6; and LCDR1 comprising or consisting of SEQ ID NO:5: Includes.

[0073] In some embodiments, the antibody or antigen-binding fragment thereof that binds to CD70 is optionally The antibody or antigen-binding fragment has the above CDR sequences and is an IgG, preferably an IgG1. In one embodiment, the antibody or antigen-binding fragment thereof that binds to CD70 has at least the sequence set forth in SEQ ID NO:4. and / or a sequence that is at least 70%, at least 80%, at least 90%, or at least 95% identical to a variable heavy domain (VH domain) consisting of or at least 70% identical to SEQ ID NO:8; or comprising a sequence which is at least 80%, at least 90% or at least 95% identical to In some embodiments, the antibody binds to CD70 and comprises a variable light chain domain (VL domain) consisting of The antibody molecule comprises a variable heavy domain (VH domain) comprising or consisting of SEQ ID NO:4. and a variable light domain (VL domain) comprising or consisting of SEQ ID NO:8. and / or VL domains identified as having a specific percentage identity to a reference sequence. In an embodiment, the VH and / or VL domain may retain the CDR sequences of the reference sequence. The CD70 antibodies or antigen-binding fragments specified herein with respect to SEQ ID NOs: 4 and 8 are It may also have the CDR sequences represented by 1 to 3 and 5 to 7.

[0074] The CD70 antibodies or antigen-binding fragments thereof that can be incorporated into the combinations described herein are antibody-drug complexes. ADCs include, for example, auristatins and maytansine or other cytotoxic agents. Some ADCs use antibody blocking and / or While maintaining effector function (e.g., ADCC, CDC, ADCP), the conjugated The anti-CD70 ADC delivers the induced active agent to cells expressing a target (e.g., CD70). , borsetuzumab mafodotin (also known as SGN-75, Seattle Genetics), SGN-70A (Seattle Genetics), tle Genetics), and MDX-1203 / BMS936561 (Bristol-Myers Squibb), can be used in accordance with the present invention. Suitable anti-CD70 ADCs are also described in WO2008074004 and WO20 No. 04073656, each of which is incorporated herein by reference.

[0075] (Venetoclax) In some preferred embodiments of the invention, the CD70 antibodies or antigen-binding The combination fragment is combined with venetoclax or a pharma- ceutical acceptable salt thereof. Clax, as described herein, is a small molecule inhibitor of BCL-2.

[0076] Venetoclax for use in the combination therapy described herein is intended to be effective in treating BCL-2 tachycardia. The compound may be provided in any suitable form that effectively inhibits protein synthesis. Such forms include, but are not limited to, Any suitable polymorphic, amorphous or crystalline form, or any In some embodiments, the compounds described herein include any isomer or tautomeric form. The combination therapy can be carried out according to the methods described in US2010 / 0305122, which is incorporated herein by reference. In another embodiment, the compound is a venetoclax compound synthesized according to the process described herein. The combination therapy described above is disclosed in EP3333167, WO2017 / 156398, WO2018 / 029711, CN107089981(A), WO201 8 / 069941, WO2017 / 212431, WO2018 / 009444, CN107648185(A), WO2018 / 167652, WO2018 / 15 No. 7803, CZ201769, both of which are incorporated herein by reference. In some embodiments, the compound is venetoclax in the form or synthesized according to the process described above. In the present specification, the combination therapy is described in WO2012 / 071336 (incorporated herein by reference). The drug contains venetoclax in any of the crystalline or salt forms described herein.

[0077] Pharmaceutically acceptable salts for use in accordance with the present invention include salts of acidic or basic groups. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, bromide, Hydrogen salts, hydroiodides, nitrates, sulfates, bisulfates, phosphates, acid phosphates, isophosphates Nicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate , bitartrate, ascorbate, succinate, maleate, gentisinate sinate, fumarate, gluconate, glucaronate, sugar salt, formate, Benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate p-toluenesulfonate, p-toluenesulfonate and pamoate (i.e. 1,1'-methylene-bis-(2-hydro Pharmaceutically acceptable salts are formed with various amino acids. Suitable base salts include, but are not limited to, aluminum salts. Salt, calcium salt, lithium salt, magnesium salt, potassium salt, sodium salt, zinc salt, and diethanolamine salts.

[0078] Venetoclax for use in the combination therapies described herein may be administered in the form of a hydrate, anhydrous or Alternatively, it may be provided in the form of a solvate. Venetoclax is marketed by AbbVie Inc. and Genentech under the trademark Venclexta. In some embodiments, the compositions described herein are commercially available under the trade name The combination includes an antibody or antigen-binding fragment thereof that binds to CD70 and Venclexta®. nothing.

[0079] (Additional Medications) The combinations of the invention may include one or more additional agents, such as one or more additional anti-cancer agents. Cut. In some embodiments, the combination comprises one or more “nucleoside metabolic inhibitors” (NMIs). NMI includes epigenetic modifications (e.g., methylation, demethylation) of nucleotides (DNA and / or RNA). Nucleoside metabolism inhibitors are molecules that block the acetylation, acetylation, or deacetylation of nucleosides. Examples of inhibitors include hypomethylating agents (HMAs), isocitrate dehydrogenase (IDH) inhibitors, and histone dehydrogenase inhibitors. Acetylase (HDAC) inhibitors and bromodomain and extraterminal (BET) inhibitors Preferred nucleoside metabolism inhibitors are hypomethylating agents. , which inhibit the normal methylation of DNA and / or RNA. Examples of hypomethylating agents are azacytidine, Decitabine and guadecitabine.

[0080] In a preferred embodiment, the combination of the present invention additionally contains azacitidine (as defined herein). Thus, in a preferred embodiment, The present invention relates to (i) an antibody or an antigen-binding fragment thereof that binds to CD70; (ii) venetoclax or an antigen-binding fragment thereof. a pharma- ceutically acceptable salt; and (iii) azacitidine.

[0081] In a further preferred embodiment, the combination of the invention additionally comprises decitabine. In a preferred embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to CD70; (ii) venetoclax or a pharma- ceutical acceptable salt thereof; and (iii) a combination comprising decitabine. Provide a service.

[0082] Azacitidine is an analog of cytidine and decitabine is its deoxy derivative. Azacitdine and decitabine downregulate gene expression through promoter hypomethylation Inhibition of DNA methyltransferases (DNMTs), which are known to upregulate Such hypomethylation disrupts cellular functions and results in cytotoxic effects. I agree.

[0083] In some embodiments, the combination of the present invention additionally comprises cytarabine. Ara-C (also known as "cytosine arabinose" or "ara-C") is a commonly used treatment for AML. High-dose cytarabine is a chemotherapy drug commonly used in newly diagnosed patients with AML. It forms part of the "7+3" standard induction chemotherapy regimen used in patients with advanced ovarian cancer. Low-dose cytarabine is It may also be used in AML patients who are not eligible for induction chemotherapy. Tarabin in combination with venetoclax is a new treatment for patients ineligible for standard induction chemotherapy The combination of the present invention is preferably administered to patients with AML who have been treated with chemotherapy or chemotherapy alone. This can be done.

[0084] In some embodiments, the combination of the present invention comprises an additional anti-cancer agent. The cancer drug is selected from any drug suitable for the treatment of myeloid malignancies, preferably AML. Preferred agents include: selectin inhibitors (e.g., GMI-1271); FMS-like tyrosine kinase inhibitors (e.g., tyrosine kinase inhibitors); Floxacin kinase receptor 3 (FLT3) inhibitors (e.g., midostaurin or gilteritinib); Phospho-dependent kinase inhibitors; Aminopeptidase inhibitors; JAK / STAT inhibitors; Cytarabine; Fludarabine; anthracycline compounds (e.g., daunorubicin, idarubicin); Xorubicin; Hydroxyurea; Vyxeos; IDH1 or IDH2 inhibitors, IDH Idhifa (or Enasidenib) or Tibsovo (or Ivosidenib), etc.; moothened inhibitors, glasdegib, etc.; BET bromodomain inhibitors; CD123 or CD33 targeting agents; HDAC inhibitor; LSC targeting agent; AML bone marrow niche targeting agent; NEDD8 activating enzyme inhibitor, Pebonedi Stats, etc.; G-CSF, as well as topoisomerase inhibitors, mitoxantrone, selinexon The drug may be selected from among cyclosporine, etoposide, and the like.

[0085] Combination Formulations The combination of drugs described herein can be administered to a subject or patient in need thereof, preferably The combination or administration may be carried out in any manner that allows for the combination therapy to be administered to a human subject or patient in need thereof. The combination can be formulated for single dose administration or for multiple dose administration. It can be made into a In some embodiments, the agents of the combination are co-formulated, i.e., in a single pharmaceutical composition. In embodiments where the agents are co-formulated, the combination or composition may be This makes it suitable for simultaneous administration of drugs.

[0086] In a preferred embodiment, the agents of the combination described herein are administered as separate compositions or medicaments. In embodiments where the agents are formulated separately, different agents or The compositions may be administered simultaneously or separately. Different compositions may be administered separately. When administered sequentially, the agents may be administered in any order. The interval between administration of the agents may be any The administration of the different compositions may be done at any suitable time interval. may be performed repeatedly (for multiple dose administration).

[0087] The CD70 antibodies or antigen-binding fragments of the combinations described herein may be administered in any suitable pharmaceutical carrier, including, but not limited to, azidopropyl ether, acetaminophen ... The antibody may be formulated with adjuvants and / or excipients. Agent technology is well known in the art and is described, for example, in Wang et al. (2007) Journal of Pharmacology. The results are reviewed in Maceutical Sciences, 96:1-26, the contents of which are cited in their entirety. The present invention can be used to formulate the antibody composition. Pharmaceutically acceptable excipients include, but are not limited to, ion exchangers, arsenic, phosphate buffers, etc. mina, aluminum stearate, lecithin, serum proteins, e.g. human serum albumin Buffer substances, e.g., phosphate, glycine, sorbic acid, potassium sorbate, saturated Physical properties: Partial glyceride mixture of fatty acids, water, salt or electrolytes, e.g. protamine sulfate, phosphate Disodium hydrogen, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, Magnesium trisilicate, polyvinylpyrrolidone, cellulose-based materials (e.g., cal Sodium carboxymethylcellulose), polyethylene glycol, polyacrylate, wax Polyethylene-polyoxypropylene-block polymer, polyethylene glycol This includes wool and wool fat.

[0088] The BCL-2 inhibitor (preferably venetoclax or a pharma- ceutical acceptable salt thereof) may be administered The composition may be formulated using any suitable pharmaceutical carrier, adjuvant and / or excipient. Suitable agents include, for example, encapsulating materials or absorption enhancers, antioxidants, binding agents, buffering agents, co- Coating agents, coloring agents, diluents, disintegrants, emulsifiers, bulking agents, fillers, flavoring agents, moisturizing agents agents, lubricants, flavorings, preservatives, propellants, releasing agents, sterilizing agents, sweeteners, solubilizing agents, wetting agents and others This includes additives such as mixtures of these.

[0089] In some embodiments, the composition is administered to a subject via any suitable route. The compositions are formulated for administration, including but not limited to, intramuscularly, intravenously, Intradermal, intraperitoneal injection, subcutaneous, epidural, nasal, oral, rectal, topical, inhalation, buccal (e.g. In some embodiments, the composition is administered in an aqueous solution. The composition may be formulated as a tablet, capsule, powder, or any other suitable dosage form.

[0090] Preparation of a composition comprising a BCL-2 inhibitor (preferably venetoclax) for oral administration in a solid dosage form Examples of excipients for the preparation include agar, alginic acid, aluminum hydroxide, benzyl alginate, etc. Cole, benzyl benzoate, 1,3-butylene glycol, carbomer, castor oil, cellulose cellulose acetate, cocoa butter, corn starch, corn oil, cottonseed oil, crospovidone , diglyceride, ethanol, ethyl cellulose, ethyl laureate , ethyl oleate, fatty acid ester, gelatin, germ oil, glucose, glycerol, Groundnut oil, hydroxypropyl methylcellulose, isopropanol, isotonic salt Water, lactose, magnesium hydroxide, magnesium stearate, malt, mannitol, mono Glycerides, olive oil, peanut oil, potassium phosphates, potato starch, povidone , propylene glycol, Ringer's solution, safflower oil, sesame oil, carboxymethylcellulose sodium Thorium, Sodium Phosphate, Sodium Lauryl Sulfate, Sodium Sorbitol, Soybean oil, stearic acid, stearyl fumarate, sucrose, surfactant, talc, tragacanth , tetrahydrofurfuryl alcohol, triglycerides, water, and mixtures thereof. A composition comprising a BCL-2 inhibitor (preferably venetoclax) administered orally in a liquid dosage form. Excipients for the preparation of the compound include, for example, 1,3-butylene glycol, castor oil, corn oil, cottonseed oil, etc. Seed oil, ethanol, sorbitan fatty acid ester, germ oil, soybean oil, glycerol, isoprene Ropanol, olive oil, polyethylene glycol, propylene glycol, sesame oil, water and mixtures thereof. Excipients for the preparation of compositions containing cyclohexyl ether (Clax) include, for example, chlorofluorohydrocarbons, Ethanol, water and mixtures thereof. Parenterally administered BCL-2 inhibitors (preferably Excipients for the preparation of compositions containing venetoclax include, for example, 1,3-butanediol, oleic acid, castor oil, corn oil, cottonseed oil, glucose, germ oil, soybean oil, liposomes, oleic acid , olive oil, peanut oil, Ringer's solution, safflower oil, sesame oil, soybean oil, United States Pharmacopoeia (USP) or include isotonic sodium chloride solution, water and mixtures thereof.

[0091] In embodiments where the agents of the combination are formulated separately, i.e., as separate compositions, The separate compositions may be formulated for administration by the same route. In embodiments where the agents are formulated separately, i.e., as separate compositions, the separate compositions The products can also be formulated for administration by different routes. For example, the CD70 antibody or antigen The binding fragment may be formulated for intravenous administration and administered in combination with a BCL-2 inhibitor, preferably venetoclax. ) may be formulated for oral administration.

[0092] As mentioned above, the combination therapy of the present invention can include Venclexta®. Neclexta® is marketed and sold by AbbVie Inc. and Genentech. Venetoclax is a product of Venclexta (registered trademark) for oral administration. It is available as a pale yellow or beige tablet containing 10, 50, or 100 mg of venetoclax. Each tablet also contains the following inactive ingredients: copovidone, colloidal silicon dioxide, polyisoprene, The ingredients include carbamide 80, sodium stearyl fumarate, and calcium phosphate dibasic. The 10 mg and 100 mg coated tablets contain the following: yellow iron oxide, polyvinyl alcohol, The ingredients are talc, polyethylene glycol, titanium dioxide, and 50 mg of coating. The tablets also contain the following: yellow iron oxide, red iron oxide, black iron oxide, polyvinyl alcohol, Contains talc, polyethylene glycol and titanium dioxide. CD70 antibody or antigen binding fragment In one embodiment, the antibody or antigen-binding fragment of the CD70 antibody is a combination of the antibody and Venclexta®. may be formulated for intravenous administration, while Venclexta® may be administered in tablet form as described above. It may also be provided as one or more.

[0093] In addition to the CD70 antibody or antigen-binding fragment and the BCL-2 inhibitor (preferably venetoclax), The combinations of the present invention, which comprise or consist of one or more additional agents, may be used in combination with other It may be formulated for administration by the same route or a different route compared to the agent. For example, The combination comprises (i) an antibody or antigen-binding fragment thereof that binds to CD70; and (ii) venetoclax or its derivatives. Pharmacologically acceptable; and (iii) in a preferred embodiment, including azacitidine, The antibody or antigen-binding fragment may be administered intravenously, and the venetoclax or pharmaceutical thereof may be administered intravenously. The corresponding acceptable salts may be administered orally, while azacitidine may be administered by subcutaneous injection. The combination comprises (i) an antibody or antigen-binding fragment thereof that binds to CD70; and (ii) venetoclax or its derivatives. Pharmacologically acceptable; and (iii) in a preferred embodiment comprising decitabine, The antibody or antigen-binding fragment may be administered intravenously, and venetoclax or a pharmaceutical thereof may be administered intravenously. The acceptable salts may be administered orally, while decitabine may be administered by subcutaneous injection.

[0094] (C.Treatment method) The combination therapy described according to the first aspect of the present invention is useful for the treatment of malignant tumors, particularly bone marrow tumors, in human subjects. The compounds can be used in methods for treating malignant tumors.

[0095] The present invention relates to a method for the treatment of malignancies, particularly myeloid malignancies, in human subjects, comprising administering to said subjects a therapeutically effective amount of CD40 or CD45 or CD45 inhibitors. 70 or an antigen-binding fragment thereof, which is administered in combination with a BCL-2 inhibitor. The present invention also provides a method for treating malignancies, particularly myeloid malignancies, in a human subject, or an antigen-binding fragment thereof. A BCL-2 inhibitor for use in the treatment of tumors, comprising an antibody that binds to CD70 or an antibody thereof. The present invention provides a BCL-2 inhibitor administered in combination with an original binding fragment.

[0096] In a preferred embodiment, the BCL-2 inhibitor is venetoclax or a pharma- ceutical acceptable salt thereof. It is a salt that can be used. The present invention further provides a method for the treatment of malignancies, particularly myeloid malignancies, in a human subject. According to a first aspect of the present invention there is provided a combination.

[0097] In a further aspect, the present invention provides a method for treating malignancies, particularly myeloid malignancies, in a human subject. The method comprises administering to a subject a combination according to the first aspect of the invention. The present invention also provides a method for treating a malignancy, particularly a myeloid malignancy, in a human subject, comprising: (i) administering to a subject a therapeutically effective amount of a compound selected from the group consisting of 1) a medicament for treating a myeloid malignancy; (ii) administering to the subject an antibody or antigen-binding fragment thereof that binds to CD70; and Administering a medicament for the treatment of atherosclerosis with a β-blocker-2 inhibitor, preferably venetoclax or a pharma- ceutical acceptable salt thereof. The method includes steps (i) and (ii) of the method, which may be performed in any order. Good too.

[0098] All of the embodiments described above with respect to the combination of the first aspect of the invention are equally applicable to the present invention. The method described in detail herein can be applied. The term "malignant tumor" refers to a disease in which abnormal cells grow in an uncontrolled manner and invade surrounding tissues. Malignant cells that enter the body's blood and lymphatic systems migrate to distant sites in the body, In some embodiments, the methods described herein include a method for determining whether a patient is capable of metastasizing to a second site. and the production of cancer progenitor or stem cells expressing CD70, CD27, or both. As described herein, upregulated CD70 expression is a therapeutic option for renal cell carcinoma. in different types of cancer, including breast cancer, metastatic breast cancer, brain tumors, leukemia, lymphoma and nasopharyngeal carcinoma Co-expression of CD70 and CD27 is also detected in acute lymphoblastic lymphoma and T-cell lymphoma. In some embodiments, the method of the present invention is described herein. The described methods include treating any of the aforementioned malignancies associated with CD70 expression, CD27 expression, or both. Regarding the treatment of.

[0099] In certain embodiments, the methods described herein are for treating myeloid malignancies. where myeloid malignancies refer to any clonal disorder of hematopoietic stem or progenitor cells. The myeloid malignancies treated according to the methods of the present invention are newly diagnosed myeloid malignancies. The patient may have relapsed / refractory myeloid malignancies.

[0100] In some embodiments, the myeloid malignancy is acute myeloid leukemia (AML); myeloid dysplasia Myeloproliferative Neoplasms (MPN); Chronic Myelogenous Leukemia (CML); and Chronic Myelomonocytic Leukemia (CML). In a preferred embodiment, the myeloid malignancy is selected from acute myeloid leukemia (CMML). The disease is AML.

[0101] Myeloid malignancies are classified according to the WHO 2008 classification combined with the 2016 revision to this classification. and the like, which are incorporated herein by reference, and which can be specifically diagnosed using the methods described in Arber et al. Please see reference (2016) Blood, 127(20):2391-2405.

[0102] Acute myeloid leukemia (AML) refers to a hematopoietic neoplasm involving bone marrow cells. AML is characterized by impaired mitochondrial function. AML is characterized by the clonal expansion of myeloid precursors with the ability to differentiate into myeloid cells. Patients with AML have blast cells in the bone marrow. Blast cells also accumulate in the peripheral blood of AML patients. The diagnosis is made when a patient has 20% or more blast cells in the bone marrow or peripheral blood.

[0103] According to the WHO classification, AML is generally classified into the following subtypes: AML with recurrent genetic abnormalities; AML with myelodysplasia-related changes; Therapy-related myeloid neoplasms; Myeloid sarcoma; Down's syndrome-related myeloproliferation blastic plasmacytoid dendritic cell neoplasm; and AML, unclassified (e.g., acute megakaryoblastic leukemia, disease, acute basophilic leukemia).

[0104] AML can also be classified according to the French-American-British (FAB) classification, which is Subtype: M0 (acute myeloblastic leukemia, minimally differentiated); M1 (acute myeloblastic leukemia, anaplastic M2 (acute myeloblastic leukemia, granulocytic mature type); M3 (promyelocytic leukemia, or acute Promyelocytic leukemia (APL); M4 (acute myelomonocytic leukemia); M4eo (myelomonocytic, bone marrow eosinophilic M5 (acute monoblastic leukemia (M5a) or acute monocytic leukemia (M5b)); M6 (acute erythroleukemia leukemia, including erythroleukemia (M6a) and rare pure erythroleukemia (M6b); or M7 (acute megakaryoblastic leukemia) Includes.

[0105] As used herein, "AML" refers to AML according to the WHO classification and / or FAB classification, unless otherwise specified. Some AML subtypes have a relatively good prognosis. Some are considered to have an intermediate prognosis, and some are considered to have a poor prognosis. Know which subtypes may be in the at-risk category.

[0106] Myelodysplastic syndromes (MDS) are characterized by dysplasia, cytopenias, and / or decreased bone marrow cellularity and and / or characterized by abnormal changes in myeloid cell differentiation, e.g., increased blast cell infiltration According to the WHO classification, MDS is generally divided into the following subtypes: MD with single lineage dysplasia; S (previously called "refractory cytopenia with monolineage dysplasia"; refractory anemia, refractory Myelodysplastic syndromes (MDS) with ringed sideroblasts, monolineage Includes the subtypes with vascular dysplasia and the subtypes with polycythemia gravis (previously known as refractory anemia with cytoplasmic sideroblasts); MDS with polycytoplasmic dysplasia (previously called polycytoplasmic sideroblasts) Myelodysplastic syndrome (MDS-EB, hereafter referred to as "refractory cytopenia with blast lineage dysplasia"); (Previously called "refractory anemia with excess blasts"), which is based on the blast percentage, It can be further subdivided into MDS-EB-1 and MDS-EB-2; MDS with isolated deletion (5q); and Includes uncontrolled MDS.

[0107] MDS can also be classified according to the French-American-British (FAB) classification, which Subtypes: M9980 / 3 (refractory anemia (RA)); M9982 / 3 (refractory anemia with ringed sideroblasts (RARS)); M9983 / 3 (refractory anemia with excess blasts (RAEB)); M9984 / 3 (refractory anemia with excess blasts in transition RAEB-T (Recurrent myelomonocytic leukemia-T)); and M9945 / 3 (chronic myelomonocytic leukemia (CMML)).

[0108] As used herein, "MDS" refers to the WHO classification and / or the FAB classification, unless otherwise specified. For both AML and MDS, the term "AML-MDS" refers to any of the conditions encompassed by the WHO Categorization by taxonomy is preferred.

[0109] Myeloproliferative neoplasms (MPNs) are similar to MDS, but according to the WHO classification, MPNs are generally classified as follows: Subtypes below: Chronic myeloid leukemia (CML); Chronic neutrophilic leukemia (CNL); Polycythemia vera (PV); Primary myelofibrosis (PMF); Essential thrombocythemia (ET); Chronic eosinophilic leukemia, not otherwise specified ; and unclassifiable MPN.

[0110] Chronic myelomonocytic leukemia (CMML) and atypical chronic myelogenous leukemia (aCML) are classified as M according to the WHO classification. They fall under the category of DS / MPN disorders because they have overlapping clinical features between MDS and MPN. This is because it represents a bone marrow tumor with characteristic features, laboratory findings, and morphological characteristics.

[0111] Patient characterization The patients or subjects to be treated according to the methods described herein, particularly AML patients or subjects, may be newly Patients may have previously diagnosed, relapsed or primary refractory disease.

[0112] The standard approach for treating newly diagnosed AML patients is high-dose cytarabine. 7 days followed by 3 days of anthracycline (e.g., daunorubicin or idarubicin) The "standard 7+3 intensive chemotherapy" approach is characterized by the administration of is administered with the goal of inducing complete remission in AML, typically after successful chemotherapy. with the intention to undergo stem cell transplantation.

[0113] Standard intensive chemotherapy is associated with significant toxicity and side effects, which may make it difficult to tolerate these effects. These patients are not suitable for treatment with standard intensive chemotherapy. Patients may be ineligible for standard intensive chemotherapy, e.g. For example, patients may present with one or more comorbid conditions that make the toxicity intolerable, or may be unable to identify those conditions. Prognostic factors that indicate poor outcome with standard intensive chemotherapy may be indicative. The determination of an individual patient's eligibility for potential intensive chemotherapy is based on the individual patient's medical history and clinical guidelines. (See, e.g., the National Comprehensive Cancer Network (NCCN) Guidelines, which are incorporated herein by reference. This will be done by clinicians taking into consideration the following: The investigator should consider other factors, including the cytogenetic and / or molecular abnormality of the AML being treated. Patients with associated causes are often deemed ineligible for standard intensive chemotherapy.

[0114] Patients who are ineligible for standard intensive chemotherapy may instead be offered lower-intensity chemotherapy such as low-dose cytarabine (LDAC). Patients who are ineligible for standard intensive chemotherapy and are not eligible for LDAC Patients receive best supportive care (BSC), including hydroxyurea (HU) and transfusion support. can.

[0115] Patients or subjects treated according to the methods described herein are those who are "unresponsive to standard intensive chemotherapy" The combination of the present invention may be classified as "eligible for treatment with steroids" or as "eligible for treatment with steroids" due to the following: Thus, any of the above-identified therapies may be expected to be effective in treating Patients who are deemed ineligible for standard intensive chemotherapy due to reasons such as It is possible.

[0116] As mentioned above, venetoclax is not recommended for patients aged 75 years or older or with comorbidities that preclude the use of intensive induction chemotherapy. Azacitidine, decitabine or low-dose rituximab for the treatment of newly diagnosed AML in adults with It has been approved in the United States for use in combination with cytarabine at low doses. In particular embodiments, the BCL inhibitor is venetoclax or a pharma- ceutically acceptable salt thereof. In an embodiment, the patient or subject treated according to the methods described herein is 75 years of age or older. In a further embodiment, the method described herein is for the treatment of a newly diagnosed AML patient. The patient or subject to be treated according to the present invention has a comorbidity that precludes the use of intensive induction therapy. , newly diagnosed AML patients. Have comorbidities that preclude the use of intensive induction chemotherapy. Patients must meet at least one of the following criteria: Performance status 2-3, severe cardiac or pulmonary comorbidity, moderate liver impairment Such embodiments can be classified based on the following: When the BCL-2 inhibitor in the combination is venetoclax or a pharma- ceutical acceptable salt thereof, Particularly preferred.

[0117] Patients or subjects treated according to the methods described herein may be receiving other treatments, such as standard Although patients may be eligible for intensive chemotherapy, alternative treatment options include the use of the combinations described herein. For example, a patient or subject treated according to the methods described herein may receive a combination therapy. Elephants may be newly diagnosed AML patients who are also eligible for standard intensive chemotherapy.

[0118] (Additional Therapeutic Agents) The methods described herein may include administration of one or more additional therapeutic agents, such as an additional anti-cancer agent. In some embodiments, the method includes the step of administering to a subject a subject of the present invention a method for the treatment of a myeloid malignancy. This includes the administration of one or more agents for use in the treatment of AML, such as agents suitable for use in the treatment of AML. Such agents include, but are not limited to: selectin inhibitors (e.g., GMI-1271 FMS-like tyrosine kinase receptor 3 (FLT3) inhibitors (e.g., midostaurin or gilteritin) Litinib); Cyclin-dependent kinase inhibitors; Aminopeptidase inhibitors; JAK / STAT inhibitors anti-inflammatory drugs; cytarabine; fludarabine; anthracycline compounds (e.g., daunorubicin, Idarubicin; Doxorubicin; Hydroxyurea; Vyxeos; IDH1 or IDH 2 inhibitors, Idhifa (or Enasidenib) or Tibsovo (or Ivosidenib, etc.; Smoothened inhibitors, Glasdegib, etc.; BET bromodomain inhibitors, CD123 or CD33 targeting agents; HDAC inhibitors; LSC targeting agents, AML bone marrow niche targeting agents; NEDD8 activating enzyme inhibitors, such as pevonedistat; G-CSF, and topoisomerase inhibitors, such as mitoxant These include ron, selinexor and etoposide, etc.

[0119] In a preferred embodiment, the methods described herein involve the use of a nucleoside metabolic inhibitor, preferably The administration of a further agent which is a hypomethylating agent or a methylating agent is particularly preferred. Cytidine and decitabine. As mentioned above, in some embodiments, (i) a CD70 antibody or an antigen-binding fragment thereof; and (ii) a BCL-2 inhibitor, preferably venetoclax. The combinations of the present invention, which include the agonist or pharma- ceutical amines or pharma- ceutical acceptable salts thereof, may also be used in combination with an additional agent, e.g. It may also be formulated to contain cytidine or decitabine.

[0120] Alternatively, the combination comprises: (i) an antibody or antigen-binding fragment that binds to CD70; and (ii) a BCL-2 inhibitor. In an embodiment, preferably comprising venetoclax or a pharma- ceutical acceptable salt thereof, In one embodiment, the method of administering the combination to a subject further comprises administering an additional agent, e.g., azacitidine or It may also include the further step of administering decitabine. The present invention relates to a method for treating myeloid malignancies, preferably AML, in a human subject, comprising administering to said subject (i) an antibody or antigen-binding fragment thereof that binds to CD70; (ii) a BCL-2 inhibitor, preferably a venom inhibitor; (iii) netoclax or a pharma- ceutically acceptable salt thereof; and (iv) azacitidine or dexamethasone. Also provided herein is a method for treating a patient having a rheumatoid arthritis, comprising administering cytabine to said patient. A combination for use in the treatment of myeloid malignancies, preferably AML, in a human subject, comprising: (ii) an antibody or antigen-binding fragment thereof that binds to CD70; (iii) a BCL-2 inhibitor, preferably a venetoclave; or a pharma- ceutically acceptable salt thereof; and (iii) azacitidine or decitabine. , is a combination including.

[0121] (Dosage) As shown in the Examples, the combinations of the present invention exhibit effective synergistic treatment of AML cells - i.e. The level of inhibition induced by this combination is greater than the additive effect of the monotherapies alone.

[0122] Methods for determining synergistic interactions are known to those of skill in the art and are described in the Examples. The preferred method for determining whether a response has occurred is the Chou-Talalay method (Chou, TC., Cancer Res. es. 2010 Jan 15;70(2):440-6, incorporated herein by reference).

[0123] In the Chou-Talalay method, CI<1 indicates synergistic effects, CI=1 indicates additive effects, and CI>1 As shown in FIG. 1, the presence and efficacy of synergy resulting from the combination of the present invention is The range of the combination varies according to the strength of the inhibitory effect of the combination. , depending on the total concentration of the combination.

[0124] Thus, in a preferred embodiment of all aspects of the invention, a CD70 antibody or an antigen thereof is The dose at which the binding fragment is administered and / or provided in the combination, and the dose at which the BCL-2 inhibitor is administered and / or provided in the combination The doses administered and / or provided in combination are each such that the combination provides synergistic therapy. The combinations are selected so that they exhibit a CI of less than 1 as determined by the Chou-Talalay method. Preferred dosages are those in which the combination exhibits a CI of less than 0.5.

[0125] Preferably, in some embodiments, a CD70 antibody or antigen-binding fragment thereof is used in the present combination. and if a BCL-2 inhibitor is administered and / or provided in the combination. or the doses provided are those for which the combination has a CI of less than 1 and an Fa of >0 as determined by the Chou-Talalay method. Each is selected to indicate .5.

[0126] As shown in this example, synergistic effects were also observed with anti-CD70 antibody (ARGX-110), BCL-2 inhibitor (Benefit This was observed with the combination of tuximab (toclax) and HMA (decitabine). In some preferred embodiments, when the combination includes an HMA, the CD70 antibody or its antigen binding agent is The dose at which the combined fragment is administered and / or provided in the combination, the dose at which the BCL-2 inhibitor is administered and / or provided in the combination, and / or provided doses, and HMAs administered and / or provided in the combination The dosages are each selected so that the combination provides a synergistic therapeutic effect.

[0127] CD70 antibodies, particularly ARGX-110, are effective at relatively low doses in the treatment of myeloid malignancies, especially AML. Therefore, in some embodiments of all the methods of the present invention, the CD70 antibody is The antibody or antigen-binding fragment thereof may be administered in a range of 0.1 mg / kg to 25 mg / kg per dose, for example 0.1 mg / kg In some embodiments, the CD70 antibody or its The antigen-binding fragment is administered at a dose ranging from 1 mg / kg to 20 mg / kg per dose. Ranges listed include the endpoints of the range unless otherwise stated. For example, doses ranging from 0.1 to 25 mg / kg The administration included a 0.1 mg / kg dose and a 25 mg / kg dose, and included two endpoints: This also includes administration at all doses between.

[0128] In some embodiments of the methods of the invention, the CD70 antibody or antigen-binding fragment thereof is administered at a concentration of 0.1 In some embodiments, the CD70 antibody or The antigen-binding fragment is administered at a dose ranging from 0.5 mg / kg to 2 mg / kg. In embodiments, the CD70 antibody or antigen-binding fragment thereof is administered at a dose of 1 mg / kg, 3 mg / kg, 10 mg / kg, or In some preferred embodiments, the CD70 antibody or its In some preferred embodiments, the antigen-binding fragment is administered at a dose of 1 mg / kg. The CD70 antibody or antigen-binding fragment thereof is administered at a dose of 10 mg / kg.

[0129] In some embodiments, administration of multiple doses of the CD70 antibody or antigen-binding fragment is performed. In some such embodiments, each of the CD70 antibodies or antigen-binding fragments thereof The doses are spaced 10 to 20 days apart, optionally 12 to 18 days apart. Each dose of anti-CD70 antibody is administered 14 to 17 days apart.

[0130] The BCL-2 inhibitor of the present combination, preferably venetoclax or a pharma- ceutical acceptable salt thereof AM may be administered according to any regimen in which the compound has been determined to be effective. The FDA-specified information for the use of Venclexta® in L-cell therapy includes information regarding the dose escalation period. The company has proposed a dosing schedule with a 5-mg dose of 100 mg ... When zacitidine or decitabine is prescribed in combination with zacitidine or decitabine, the dosing schedule is: 100 mg of Venclexta® on day 1; 200 mg of Venclexta® on day 2. ) on day 3; 400 mg of Venclexta® on day 3; and 400 mg daily thereafter. Venclexta® 75 mg / m 2 of azacitidine or 20 mg / m 2 in combination with decitabine It is recommended that treatment consist of administering the drug until disease progression or unacceptable toxicity is observed. When Venclexta is prescribed in combination with low-dose cytarabine, the dosing schedule should be adjusted accordingly. The doses were: 100 mg of Venclexta® on day 1; 200 mg of Venclexta® on day 2. on day 3, 400 mg of Venclexta; and on day 4, 400 mg of Venclexta; and 600 mg of Venclexta® daily at 20 mg / m 2 In combination with It is recommended that treatment with serotonin should consist of administering the drug until minimal toxicity is observed.

[0131] In some embodiments, the individual Individual dosages, for example oral dosages, range from 100 mg to 600 mg. In some embodiments, the venetoclax or a pharma- ceutical acceptable salt thereof is administered at a dose of 400 mg per day. In some embodiments, venetoclax or a pharma- ceutical acceptable salt thereof is administered in an amount The venetoclax salt is administered at a dose of 600 mg per day. Daily fixed dose administration may be preceded by a titration period of, for example, 3 days, and During the maintenance period, patients will receive increasing doses of venetoclax until they reach the daily maintenance dose. Administer to.

[0132] The combination includes a nucleoside metabolic inhibitor or the method includes administering a nucleoside metabolic inhibitor. In an embodiment of the invention comprising administering to a subject a nucleoside metabolic inhibitor, the nucleoside metabolic inhibitor is administered in an amount of 20 to 100 mg / day. 00 mg / m 2 As already mentioned, the ranges described herein may be administered in the form of a dose of includes range endpoints unless otherwise specified. For example, 20 to 100 mg / m per day 2 Dosage range of Dosage is 20 mg / m per day. 2 Dosage and administration: 100 mg / m per day 2 Including administration in doses, This includes administration of all doses between the two endpoints.

[0133] In some embodiments, the nucleoside metabolic inhibitor is azacitidine and or 70-80 mg / m 2 In some preferred embodiments, the nuclease is administered in a dose range of The cleoside metabolic inhibitor was azacitidine, 75 mg / m per day. 2 is administered at a dose of

[0134] In some embodiments, the nucleoside metabolic inhibitor is decitabine and 15-25 mg / m 2 In some preferred embodiments, the nucleic acid is administered in a dose range of The leucine metabolic inhibitor was decitabine, 20 mg / m 2 per day. 2 is administered at a dose of

[0135] The combination of the invention comprises a nucleoside metabolism inhibitor or the method comprises a nucleoside metabolism inhibitor. In an embodiment of the invention involving administering the agent, the nucleoside metabolic inhibitor is administered for 5 to 10 days. The nucleoside inhibitor may be administered in a daily dose of between 100 mg / kg / day and 100 mg / kg / day for a dosing period. The doses are administered daily for a period of time or for a length of 5, 6, 7, 8, 9, or 10 days. In some preferred embodiments, the nucleoside metabolic inhibitor is administered at a daily dose of 7 days. The preferred nucleoside metabolic inhibitor is azacitidine. .

[0136] In some embodiments, the nucleoside metabolic inhibitor is administered at the end of one dosing period and The dosing regimen is for repeated dosing periods with an interval of 18 to 25 days between the start of each dosing period. That is, the dosing regimen is one dose of the nucleoside inhibitor administered daily ( For example, 5, 6, 7, 8, 9, or 10 days) including at least two dosing periods and The end of the treatment period and the start of the next treatment period are separated by 18, 19, 20, 21, 22, 23, 24, or 25 days. In some embodiments, at the end of one dosing period and the start of the next dosing period. are spaced 21 days apart.

[0137] In some embodiments, each dosing period is the same length (eg, 7 days). In some embodiments, at the end of each dosing period and at the start of the next dosing period There is an equal number of days between them (e.g. 21 days).

[0138] In some embodiments, the initial administered dose of the nucleoside metabolic inhibitor is or an antigen-binding fragment thereof, is administered 7 to 21 days after the first dose. In the present invention, the initial dose of the nucleoside metabolic inhibitor is In some embodiments, the nucleoside analog is administered 10 to 17 days after the first dose of the nucleoside analog. The first dose of the metabolic inhibitor is administered 14 days after the first dose of the CD70 antibody or antigen-binding fragment thereof. It is administered days later.

[0139] In some embodiments, one of the daily administered doses of the nucleoside metabolic inhibitor is The dose is administered on the same day as the dose of the antibody or antigen-binding fragment thereof. An embodiment of the method of the invention in which both a nucleoside metabolic inhibitor and an antigen-binding fragment thereof are administered In these patients, the dosing regimen of both the CD70 antibody and the nucleoside metabolic inhibitor was consistent with the anti-CD70 antibody effect. At least one of the scheduled doses is a scheduled daily dose of a nucleoside metabolic inhibitor. The day is the same as one of the doses of the nucleoside metabolic inhibitor. , the second, third, fourth, fifth, sixth or seventh day.

[0140] In some embodiments, the administered dose of the CD70 antibody or antigen-binding fragment thereof is 14 to 20 mg / day. The nucleoside metabolic inhibitors were administered in a daily dose for 17 days, with a 7-day repeat dosing period. The doses are administered according to a regimen in which the end of one dosing period and the start of the next dosing period are The first daily dose of the first dosing period is administered with an anti-CD70 antibody or its antigen binding site at a one-day interval. It is administered 14 days after the first dose of the combined fragment.

[0141] In some embodiments, one patient treatment cycle consists of 28 days, A 5-aminobutyric acid inhibitor, preferably azacitidine or decitabine, is administered starting on day 1 of the cycle. The treatment methods described herein may be administered daily for 1, 6, 7, 8, 9 or 10 days. Each treatment cycle may be a duplication of the previous treatment cycle. In some embodiments, a CD70 antibody, a BCL inhibitor (preferably venetoclax or Patients treated with azacytidine (or a pharmacologic salt thereof) were randomized to receive a 28-day subgroup of Patients were treated with cyclosporine and azacitidine was administered daily for the first 7 days of each 28-day cycle. In some embodiments, a CD70 antibody, a BCL inhibitor (preferably venetoclax or similar) Patients treated with 1,2-diaminobenzidine (a pharma- ceutical acceptable salt of 1,2-diaminobenzidine) and decitabine were administered in a 28-day cycle. Patients were treated with 28-day cycles of 100 mg / kg / day of chemotherapy and 100 mg / kg / day of chemotherapy, with decitabine administered daily for the first 5 days of each 28-day cycle. CD70 antibody, BCL inhibitor (preferably venetoclax or its drug) and and its acceptable salt) and a nucleoside metabolic inhibitor (preferably azacitidine or decididine). In embodiments where the CD70 antibody is administered on day 3 and / or day 4 of a 28-day cycle, It may be administered on day 17. In a preferred embodiment, the CD70 antibody is ARGX-110. In a highly preferred embodiment, the CD70 antibody (e.g., ARGX-110) is administered at a dose of 10 mg / kg. can be.

[0142] A further advantage of the present invention is the ability to reduce the incidence of NMI (e.g., azacitidine) following an initial period of combination therapy. Accumulated toxicity from long-term NMI treatment, e.g. For example, there is the possibility of cytopenias resulting from the effect of NMI on non-blast cell types. Therefore, tapering or ceasing the administration of NMI after the initial period may reduce the risk of such toxicity. In some embodiments, the present invention provides a method for the treatment of blast cell types. The treatment includes administering to the patient a CD70 antibody, a BCL-2 inhibitor (e.g., venetoclax), and an NMI, as described above. As a combination therapy according to any of the above embodiments, In the second stage, patients are given a CD70 antibody and a BCL-2 inhibitor (e.g., venetoclax). and NMI administered as a combination therapy, but with the dose of NMI in the second stage (maintenance therapy). is less than the dose of NMI administered in the first stage. The dose of NMI in the second stage is zero. That's fine too.

[0143] In such an embodiment, the CD70 antibody administered in the second stage (i.e., maintenance therapy) The dosage may be any dosage according to the embodiments previously described. In this case, the dosage is 0.1 mg / kg to 25 mg / kg, for example, 0.1 mg / kg to 20 mg / kg, for example, 1 mg / kg In some embodiments, the dosage ranges from 0.1 mg / kg to 20 mg / kg per dose. In some embodiments, the dosage ranges from 0.5 mg / kg to 2 mg / kg. In some embodiments, the dosage ranges from 1 mg / kg, 3 mg / kg, 10 mg / kg, In some embodiments, the dosage is 1 mg / kg. In some embodiments, the administered dosage is 10 mg / kg.

[0144] The duration of the first stage (i.e., induction therapy), the timing of transition to the second stage (i.e., maintenance therapy), The limits for tapering or completely stopping the dose of nicotine and NMI should be adjusted for each individual patient. The clinical outcomes are based on the individual patient's response to therapy and their medical history and are determined by the clinician. The following embodiments are therefore offered as non-limiting examples. In some embodiments, induction therapy involves administering to the patient a blastoma cell line containing the blastoma cell line of the patient's bone marrow and / or peripheral blood. The concentration is then adjusted to less than 10%, optionally less than 5%. In accordance with the present invention, induction therapy includes at least five NMI dosing periods, optionally at least six, seven, eight, nine, or fewer NMI dosing periods. The NMI treatment will be administered over a period of at least 10 sessions.

[0145] In some embodiments, the dose of the NMI during the maintenance period is 50 mg / m per day. 2 Exceeding or optionally 40 mg / m 2 Not to exceed 30 mg / m2 per day, optionally 2 Not to exceed 1 day at your discretion per 20 mg / m 2 In some embodiments, the dose of the NMI in the maintenance period does not exceed is zero.

[0146] As described elsewhere herein, the agents of the combination may be administered by any suitable route. Thus, administration of the agent according to the method of the present invention may be optionally The individual drugs may be administered via any suitable route, and it is not necessary for each drug to be administered via the same route. For example, In one embodiment, a CD70 antibody or antigen-binding fragment thereof is administered intravenously while a BCL-2 inhibitor (e.g., venetoclave) is administered intravenously. Alternatively, a low methylprednisolone drug such as azacitidine or decitabine may be administered orally to the patient or subject. In embodiments in which a vasopressin-releasing agent is administered, the agent is administered intravenously or subcutaneously by injection. It is also possible.

[0147] (Treatment effect) In some embodiments, the methods described herein include determining the patient's blast count, i.e., blast count. As used herein, "blast cells" or "blasts" refers to Myeloblasts or myeloid blasts are myeloid precursor cells in the bone marrow. In healthy individuals, blasts are They are not found in the peripheral blood circulation, and there are less than 5% blast cells in the bone marrow. Increased production of abnormal blast cells with disrupted differentiation potential in patients with cancer, especially those with AML and MDS. The overproduction of these abnormal blast cells may result in the accumulation of abnormal cells in the peripheral blood circulation or bone marrow or both. This can be detected by monitoring the blast count in the blood.

[0148] The proportion of blast cells in bone marrow or peripheral blood can be determined by methods known in the art, for example, by measuring the bone marrow blast ratio of a subject. Flow cytometric evaluation of cells obtained from bone marrow biopsies or peripheral blood smears or cell cytotoxicity The percentage of blasts can be determined relative to the total cells in the sample. For example, flow cytometry can be used to measure the ratio of CD45 to total cell counts. dim , S.S.C. lo w The number of cells can be used to determine the proportion of blast cells. The morphological assessment was performed to determine the number of morphologically identified blasts relative to the total number of cells in the examined field. can be used for.

[0149] In some embodiments, the percentage of blast cells in the bone marrow is reduced to less than 25%, less than 20%, e.g. In some embodiments, methods are provided in which the blastocysts in the bone marrow are reduced, for example to less than 10%. Methods are provided for reducing the percentage of spherical cells to less than 5%. In some embodiments, and reducing the percentage of blast cells in bone marrow to about 5% to about 25%, , the percentage of myeloblasts compared to the percentage of myeloblasts before carrying out the method (or treatment) The percentage of cells is reduced by more than 50%.

[0150] In some embodiments, the percentage of blast cells in the peripheral blood is reduced to less than 25%, less than 20%, e.g., In some embodiments, the method further comprises the steps of: In some embodiments, a method is provided for reducing the percentage of blast cells in a patient to less than 5%. The present invention provides a method for reducing the ratio of blast cells in peripheral blood to about 5% to about 25% in such a patient. In this case, the percentage of peripheral blast cells is compared to the percentage before the method (or before treatment). In all cases, the reduction in the percentage of peripheral blood blast cells was greater than 50%. For clinical determination of blast cell percentage, cell morphology (referred to as cytomorphology) is typically used. (also known as 'non-linear') evaluation is preferred.

[0151] In certain embodiments, the methods described herein induce a complete response. In this context, a complete response or "complete remission" is defined as: bone marrow blasts <5%; circulating blasts and Auer rods absence of blasts with cytology; absence of extramedullary disease; ANC ≥ 1.0 × 10 9 / L (1000μL); Platelet count ≧100×10 9 / L (100,000 μL); as described in Dohner et al. (2017) lood, 129(4):424-447.

[0152] The method demonstrates a complete response with platelet recovery, i.e., a platelet count >100×10 9 / L (100,000 / These methods can achieve a complete response with neutrophil recovery. , i.e., neutrophil count >1.0 × 10 9 A response of 1000 / μL can be achieved. Alternatively or additionally, the method may include culturing for 8 weeks or longer, 10 weeks or longer. for 12 weeks or longer, or for transfusion of red blood cells or platelets or both. It is possible to induce blood-dependence withdrawal.

[0153] In certain embodiments, the methods described herein include determining whether a patient has minimal residual disease or measurable disease that is negative. The cytotoxic T cell therapy induces a state of functional residual disease (MRD), as described in Schuurhuis et al. (2018) Bl See, ood, 131(12): 1275-1291. In some embodiments, the methods described herein include complete response without minimal residual disease. Effectiveness (CR MRD- ) is derived, see Dohner et al., supra.

[0154] The method can achieve or induce a partial response. In context, a partial response or partial remission is a 5% to 25% decrease in bone marrow blast percentage; and a reduction in pretreatment bone marrow blast percentage of at least 50%, including See Dohner et al., supra.

[0155] The methods described herein can extend survival. The term "survival" includes overall survival, 1-year survival, 2-year survival, and 5-year survival. The methods described herein refer to the specific disease being treated. or improve survival compared with gold standard treatments for symptoms Gold standard treatment is also called best practice, standard of care, standard of medical care or standard of care. For any given disease, different clinical practices may exist in different countries, e.g. There will be one or more gold standard treatments depending on the practice. These treatments available today are diverse and include chemotherapy, radiation therapy, stem cell transplants, and In addition, both US and European clinical guidelines recommend that patients with myeloid malignancies, e.g. For example, standard treatment for AML is described in O'Donnell et al. (2017) Journal of the National Comprehensive Cancer Network, 15(7):926-957, and the article by Dohner et al. See, (2017) Blood, 129(4):424-447, both of which are incorporated by reference. do. The methods of the present invention are more effective than patients receiving any standard treatment for myeloid malignancies. , survival time can be extended or improved.

[0156] The methods described herein may include the further step of subjecting the patient or subject to a bone marrow transplant. can. The methods described herein also include preparing a patient or subject with a myeloid malignancy for bone marrow transplantation. As described above, the method of the present invention can be used to prepare bone marrow or It can be performed to reduce the absolute or relative number of blast cells in the peripheral blood. In some embodiments, the method includes the step of: The method is carried out to reduce the incidence of bone marrow transplantation in a patient or subject. , can be used to reduce blast cell counts to less than 5%.

[0157] (D. Kit) The combinations described herein may be provided in the form of a kit packaged with instructions for use. This is also fine.

[0158] (Incorporating References) Various publications are cited in the foregoing description and throughout the examples which follow, including, but not limited to, the following: Each is incorporated herein by reference in its entirety. EXAMPLES

[0159] (Example) In a recent Phase I clinical trial, elderly, unmatched AML patients were treated with a humanized monoclonal antibody with enhanced ADCC activity. The clonal anti-CD70 antibody (mAb), cusatuzumab (also referred to herein as ARGX-110), is combined with HMA. The combination treatment has shown promising clinical activity and a favorable tolerability profile .

[0160] Venetoclax, a BCL-2 antagonist, is being studied in Phase I and II trials in combination with standard of care Inhibition of oxidative phosphorylation targets leukemia stem cells (LSCs) in elderly AML patients in clinical trials The study showed very promising activity by targeting and eliminating inflammatory cytokines (Pollyea et al., Nature Medicine (2018) 2). 4; 1859-1866). However, even with the use of new drugs such as venetoclax, refractory or recurrent disease remains. There remain some patients who are susceptible to venetoclax. It was hypothesized that combining this with cusatuzumab, which has a mechanism of action, could result in successful elimination of LSCs. I was fooled.

[0161] (method) To test this hypothesis, a drug combination study was performed using the Chou-Talalay method (Chou TC, Cancer According to the method described in Research (2010) 70(2); 440-6, CD70-expressing A cells such as MOLM-13, NB-4, and NOMO-1 cells were cultured in vitro. In vitro studies were performed on AML cell lines. MOLM-13 AML cells express FLT3-ITD and NOMO-1 Cells express t(9;11)(p22;q23). Each of these genetic abnormalities predicts patient outcome. NB-4 is an acute promyelocytic leukemia (APL) cell line, and this APL is a rare cancer in AML patients. Notably, NOMO-1 and MOLM-13 cells express mRNA and protein MOLM-13 AML cells also express BCL-2. It is expressed in the IL-1 receptor β-blocker (IL-1R) and is sensitive to BCL-2 inhibition (Lin et al., Scientific Reports (2016) 6; 27696).

[0162] MOLM-13 cells (Matsuo et al., Leukemia (1997) 11 (9): 1469-77), NOMO-1 cells (Kato (Acta Haematol Jpn, 1986), MV4-11 cells and NB4 cells (Lanotte et al., Blood (1991) 77 (5); 1080-86) were purchased from ATCC. Cell lines were tested for mycoplasma-free status. , ATCC-recommended FCS-containing medium, GlutaMAX supplement, 100 U / mL penicillin, 100 μL Cultures were grown at 37°C in a humidified atmosphere of 95% air and 5% CO2 containing 10 μg / mL streptomycin.

[0163] First, cells from each AML cell line were treated with either decitabine, cusatuzumab or venetoclave. IC for each treatment 50 It was decided. 10 5 AML cells, In the presence of CFSE-labeled NK cells from healthy individuals (1:1 ratio), cusatuzumab (0.1, 1.0 and 10 μg / m l), venetoclax (0.5 and 200 nM), a range of decitabine (0.01–1 μM), or vehicle The assay was performed in triplicate.

[0164] I C 50 Upon determination, the following working concentrations were identified for subsequent synergy experiments: (1) IC 50 , MOLM-13 cells: (decitabine: 0.01 nM, venetoclax: 0.42 nM, kusatsuma B: 0.68 μg / ml) (2) IC 50 , NOMO-1 cells: (decitabine: 0.001 nM, venetoclax: 3.4 nM, kusatsuma B: 0.14 μg / ml) (3) IC 50 , NB4 cells: (decitabine: 4.8 nM, venetoclax: 17.3 nM, cusatuzumab: 0 .3 μg / ml) (4) IC 50 , MV4-11 cells: (decitabine: 2.36 nM, venetoclax: 5 nM, cusatuzumab: 1.2 μg / ml).

[0165] To determine synergy, drugs were compared at equipotent ratios (IC501 / IC502 or IC501 / IC502 / IC503). Fixed combination ratio experiments were performed in which the drugs contributed equally to cell killing. The response was as previously described (Riether et al., Sci Transl Med (2015) 7(298); 298 ra119), and all doses / dose combinations were evaluated in duplicate technical replicates.

[0166] NOMO-1, MOLM-13, NB-4 or MV4-11 AML cell lines were treated with vehicle, decitabine, or kusatsuma. venetoclax or venetoclax alone, venetoclax and decitabine, decitabine and custard venetoclax and cusatuzumab, or a combination of two of each, or desitavi All combinations were compared in the triple combination of venetoclax, cusatuzumab, and venetoclax. High concentration and three low concentrations (specified IC 50 Test at a constant ratio of 1000 mg / kg (higher and lower concentrations than Cells were cultured in the presence of CFSE-labeled NK cells (1:1 ratio). Annexin V staining for 72 h The number of viable AML cells was then assessed, and the effect of drug treatment was assessed as the ratio of viable cells to vehicle-treated cells. The cell ratio was calculated.

[0167] Combination index (CI) was calculated using CompuSyn software and the effect ratio (Fa) value was calculated. The combination index (CI) for all combinations was plotted against the activity ratio (Fa). The results are plotted in FIG. 1, and the individual combinations are shown in FIG.

[0168] Fa values ​​of 0, 0.5, and 1 correspond to 0%, 50%, and 100% dead cells. A CI of 0.01 or less indicates synergy, additivity, and antagonism, respectively. 50 Yes, that Each IC 50 The Fa value reached for the combination of concentrations is shown. Fa-CI plot method for evaluating synergy The principles and advantages of this method are described, for example, in Chou TC, Cancer Research (2010) 70(2); 440-6 and Zhao The articles are provided in the literature by E. et al., Front Biosci (Elite ed.), (2010) 2; 241-249 (each of these (which is incorporated herein by reference in its entirety).

[0169] In addition to the above, cusatuzumab / venetoclax combination and cusatuzumab / venetoclax / H The effect of the MA combination was tested on primary LSCs from AML patients. + CD38 - leukemia stem cell Lymphocyte-specific colonies (LSCs) were isolated from newly diagnosed AML patients and treated with cusatuzumab, decitabine, or The colony-forming and repopulation potential of LSCs was then assessed. The effect on seeding potential was assessed.

[0170] In particular, CD34 from three AML patients (P1, P2, and P3) + CD38 - AML LSCs were combined with NK cells (1:1 ratio) In the presence of 0.3 μg / ml of Cusatuzumab (Cusa) or venetoclax (Ve) (6 nM), Duplicate overnight cultures were grown as single agents or in combination and then plated on methylcellulose. In addition, CD34 cells from two AML patients (P4 and P5) + CD38 -AML LSCs were cultured in the presence of NK cells (1:1 ratio). In the presence of cusatuzumab (Cusa: 0.3 μg / ml), decitabine (0.01 μM), or venetoclax (Ve : 6 nM) as single agents or in combination overnight. Colony formation was assessed after 14 days. .

[0171] (result) (1. Cusatuzumab, when used in combination with venetoclax and / or decitabine, (showed synergistic effects in eliminating AML cell lines in vitro) Venetoclax and / or decitabine in combination with cusatuzumab inhibited CD70-expressing NOMO-1 AM L cells were synergistically eliminated over a wide dose range (see Fig. 1 and Fig. 2B-D).

[0172] This combination resulted in higher efficacy levels (Fa>0.7) associated with greater tumor killing (Chou et al., 2010). Importantly, venetoclax and cusatuzumab (Ven / C) showed strong synergistic effects. usa) and venetoclax, cusatuzumab, and decitabine (Ven / Cusa / Dec) had CIs close to 0.1. Venetoclax and decitabine (Ven / Dec) and Satuzumab and decitabine (Cusa / Dec) also showed a high level of efficacy with a maximum CI of approximately 0.5. The synergistic effect of the Ven / Cusa combination was greater than that observed with the Ven / Cusa / Dec combination. The effect was similar.

[0173] Similar results were observed in NB4 and MV4-11 AML cell lines, with all cusatuzumab combinations showing high The results showed a strong synergistic effect with a low effect level (Fa>0.5) (see Figures 3 and 5).

[0174] In the MOLM-13 cell line, all two combinations showed synergistic effects at lower efficacy levels. Venetoclax / decitabine and cusatutzumab / decitabine had 0. Synergistic effects were observed only at drug concentrations below Fa of 4 and Fa of 0.6. At the efficacy level (Fa of approximately 0.7 to 0.8), cusatutzumab / venetoclax The combination showed strong synergy (see Figure 4), but some antagonism at the maximal efficacy level. The combination of venetoclax, decitabine and cusatuzumab was showed low levels of antagonism at effective levels.

[0175] (2) When used in combination with venetoclax and / or decitabine, cusatuzumab demonstrated synergistic effects in eliminating primary human AML leukemia stem cells (LSCs) in vitro To evaluate the efficacy of the cusatuzumab / venetoclax combination in primary human AML: LSCs from five AML patients (P1–P5), CD34 + CD38 - LSC was treated. Patient characteristics are shown below. vinegar.

[0176] Table 2: Characteristics of patients P1-P5 [Table 2]

[0177] The results for patients P1, P2 and P3 are shown in Figures 6 and 7. Figure 6 shows the wells formed after treatment. The absolute number of colonies per patient is shown, which is an index of LSC numbers. The same data presented as a percentage of the average number of colonies per well in the treatment group are shown.

[0178] Figures 6 and 7 show the synergy between cusatuzumab and venetoclax observed in AML cell lines. The results were manifested as a strong and significant reduction in LSC numbers compared with either treatment alone. As shown in Figures 6A and 7A. 6B and 7B show the effect of reducing LSC numbers by replating primary colonies in the absence of treatment molecules. This indicates that the trait was maintained even when bred.

[0179] The results for patients P4 and P5 are shown in Figure 8. In these patients, cusatuzumab, decitabine and The triple combination of venetoclax had the same efficacy as the combination of cusatuzumab and venetoclax This is because the triple combination showed increased synergistic effects compared to the Ven / Cusa combination (Figure 1). This is consistent with the fact that it was not observed.

[0180] 3. Venetoclax Increases CD70 Expression The effect of venetoclax on CD70 expression was evaluated at both the mRNA and protein levels The results are shown in Figures 9 and 10, and show that in the presence of venetoclax, CD70 expression was reduced in AML cells. It was clearly shown that expression was upregulated.

[0181] (Conclusion) This study will explore the use of cusatuzumab and venetoclax and / or hypomethylating agents (e.g., azathioprine). Combination therapy with cytidine or decitabine was more effective than either monotherapy alone The present study was carried out to determine whether the combination therapy could provide a more effective treatment for AML. We also explored whether it could demonstrate a therapeutic effect.

[0182] Combination index (CI) values ​​were provided by the Chou-Talalay method and indicate whether the drug combination is synergistic. is an established means to determine whether compounds interact in a potent, additive, or antagonistic manner. We hypothesized that venetoclax and cusatuzumab may act in a synergistic manner. Mechanistically, this is because venetoclax treatment upregulates CD70 on AML cells. This is because we were able to show that venetoclax can induce L This would render SCs more susceptible to cytolytic killing by cusatuzumab. , Chou Cancer Res. (2010) Jan 15; 70(2):440-6, incorporated herein by reference. The mechanism of action of each drug is described in Even with some knowledge, synergy between drugs is difficult to predict.

[0183] The data shown in Figures 1 to 5 indicate that the Ven / Cusa combination has a particularly high level of efficacy against AML cells. The strong synergistic effect was also demonstrated by the hypomethylating agent. Patients were maintained on a triple combination containing decitabine as chemotherapy.

[0184] This synergy is due to the fact that when these drugs are combined, they achieve the same effect as monotherapy. This indicates that significantly lower concentrations of each drug can be used compared to the concentrations required for This is particularly advantageous.

[0185] Cell line studies have demonstrated strong synergy between the venetoclax / cusatuzumab combination, The combination was tested in primary AML LSCs, which allows for a more rigorous and clinically relevant assessment of potential therapeutic effects. provides a relevant assessment of the role of these cells in promoting the aberrant growth characteristics of AML. That is why.

[0186] The synergistic effect of Ven / Cusa combination therapy observed in cell lines was consistent with that observed in primary AML leukemia cells from human patients. These data were consistent with the results of the study in all patient samples. The combination therapy produced somewhat significantly greater LSC colony size than either therapy alone. This data indicates a synergistic effect between the components of the Ven / Cusa combination. (Because of the small number of available primary cells, the Chou-Talalay method was used to compare It was not possible to test for synergistic effects.

[0187] The effect of cusatuzumab / venetoclax treatment on LSC function was examined by sequential replating in vitro When analyzed in a more rigorous manner by seeding experiments, the results after the combination treatments observed after the first seeding The colonization defect was maintained during subsequent reseeding. Although netoclax was not present during reseeding, it effectively attenuated LSCs and their proliferation potential. This is a case that shows a small

[0188] The triple combination, cusatuzumab / venetoclax / decitabine, is Reduced colony and reseeding capacity of primary human LSCs to the same extent as clax combination treatment Therefore, it is possible to combine cusatuzumab and venetaminib without the need to include an HMA such as decitabine. Using the clax combination, it is possible to achieve effective treatment of AML, thereby reducing the toxicity of the patient. This could reduce exposure to the law. The present application provides the following aspects of the invention. (Aspect 1) A combination comprising: (i) an antibody or antigen-binding fragment thereof that binds to CD70; and (ii) a BCL-2 inhibitor. . (Aspect 2) The method of claim 1, comprising: (i) an antibody or antigen-binding fragment thereof that binds to CD70; and (ii) a BCL-2 inhibitor. The combination of claim 1, wherein the BCL-2 inhibitor is compound (I) shown below: (chemical 1) TIFF2025000862000005.tif94170 or a pharma- ceutically acceptable salt thereof. (Aspect 3) The antibody or antigen-binding fragment thereof that binds to CD70 comprises a variable heavy domain (VH) and a variable light domain (VL). and a VL domain (VH) comprising the following CDR sequences: HCDR3 comprising or consisting of SEQ ID NO:3; HCDR2 comprising or consisting of SEQ ID NO:2; HCDR1 comprising or consisting of SEQ ID NO:1; LCDR3 comprising or consisting of SEQ ID NO:7; LCDR2 comprising or consisting of SEQ ID NO:6; and LCDR1 comprising or consisting of SEQ ID NO:5: 3. The combination of embodiment 1 or embodiment 2, comprising: (Aspect 4) The antibody or antigen-binding fragment thereof that binds to CD70 has at least 70% identity to SEQ ID NO:4. A VH domain comprising an amino acid sequence and an amino acid sequence that is at least 70% identical to SEQ ID NO:8. 4. The combination of embodiment 3, comprising a VL domain comprising the sequence (Aspect 5) The antibody or antigen-binding fragment thereof that binds to CD70 has the amino acid sequence represented by SEQ ID NO:4. and a VL domain comprising the amino acid sequence represented by SEQ ID NO: 8. 4. The combination described above. (Aspect 6) The combination according to any one of aspects 1 to 5, wherein the antibody is an IgG. (Aspect 7) The antibody according to any one of aspects 1 to 6, wherein the antibody has ADCC activity, CDC activity, or ADCP activity. combination. (Aspect 8) The combination according to any one of embodiments 1 to 7, wherein the antibody comprises a defucosylated antibody domain. (Aspect 9) The combination according to any one of embodiments 1 to 6, wherein said antibody is ARGX-110 (cusatuzumab). (Aspect 10) The antigen-binding fragment may comprise an antibody light chain variable domain (VL); an antibody heavy chain variable domain (VH); a single chain antibody (scFv); F(ab')2 fragment; Fab fragment; Fd fragment; Fv fragment; one-armed (monovalent) antibody; diabody, Triabodies, tetrabodies, and combinations, assemblies or combinations of such antigen-binding fragments. or any antigen-binding fragment formed by conjugation. The combination according to any one of aspects 1 to 9. (Aspect 11) The antibody or antigen-binding fragment thereof and the BCL-2 inhibitor are formulated as separate compositions. A combination according to any one of aspects 1 to 10. (Aspect 12) The combination may include at least one additional anti-cancer agent, preferably an agent for treating a myeloid malignancy. 12. The combination according to any one of aspects 1 to 11. (Aspect 13) The combination according to embodiment 12, wherein said anti-cancer agent is an agent for treating acute myeloid leukemia (AML). (Aspect 14) 14. The combination according to any one of embodiments 1 to 13, wherein the combination additionally comprises a hypomethylating agent. (Aspect 15) The combination according to embodiment 14, wherein said hypomethylating agent is azacytidine. (Aspect 16) The combination according to embodiment 14, wherein said hypomethylating agent is decitabine. (Aspect 17) The CD70 antibody or antigen-binding fragment thereof and the BCL-2 inhibitor are administered to a human subject with acute myeloid leukemia. each present in the combination in an amount sufficient to provide synergistic therapy when administered to 17. The combination according to any one of aspects 1 to 16. (Aspect 18) The CD70 antibody or antigen-binding fragment thereof and the BCL-2 inhibitor include NOMO-1, MOLM-13, NB4 and M It was enriched to provide synergistic cell killing when cultured with selected AML cell lines from V4-11. 18. The combination according to any one of embodiments 1 to 17, wherein each is present in the combination in a sufficient amount. (Aspect 19) 19. A combination according to any one of embodiments 1 to 18 for use in therapy. (Aspect 20) Aspects 1-1 for use in the treatment of malignancies, preferably myeloid malignancies, in a human subject. 10. The combination according to any one of claims 9. (Aspect 21) A method for treating malignancies, preferably myeloid malignancies, in a human subject, comprising administering to said subject an antibody capable of binding to CD70. an antibody or antigen-binding fragment thereof that binds to a BCL-2 inhibitor, or an antigen-binding fragment thereof. (Aspect 22) The antibody or antigen-binding fragment thereof is compound (I), (Case 2) 22. The use according to embodiment 21, wherein the compound is administered in combination with TIFF2025000862000006.tif96170 or a pharma- ceutical acceptable salt thereof. , an antibody or antigen-binding fragment thereof that binds to CD70. (Aspect 23) The subject is additionally administered azacitidine or decitabine. 23. An antibody or an antigen-binding fragment thereof for use as described in claim 22. (Aspect 24) BCL-2 Inhibition for Use in the Treatment of Malignancies, Preferably Myeloid Malignancies, in Human Subjects The BCL- 2 inhibitors. (Aspect 25) The BCL-2 inhibitor is compound (I), (3) 25. The BCL-2 inhibitor for use according to embodiment 24, which is TIFF2025000862000007.tif73170 or a pharma- ceutically acceptable salt thereof. (Aspect 26) The subject is additionally administered azacitidine or decitabine. 25. A BCL-2 inhibitor for use as described in claim 25. (Aspect 27) A method for treating a malignancy, preferably a myeloid malignancy, in a human subject, comprising administering to the subject the The method comprises administering an effective amount of a combination according to any one of claims 1 to 18. (Aspect 28) 1. A method of treating a malignancy, preferably a myeloid malignancy, in a human subject, comprising: (i) administering to the subject an antibody or antigen-binding fragment thereof that binds to CD70; and (ii) administering to the subject a BCL-2 inhibitor, preferably compound (I) or a pharma- ceutically acceptable salt thereof. administering a salt; The method comprising: (Aspect 29) The myeloid malignancy is selected from newly diagnosed or relapsed / refractory myeloid malignancies. The method of embodiment 27 or embodiment 28, (Aspect 30) The myeloid malignancies include acute myeloid leukemia (AML); myelodysplastic syndrome (MDS); myeloproliferative tumors. chronic myelogenous leukemia (CML); and chronic myelomonocytic leukemia (CMML). 30. The method according to any one of items 27 to 29. (Aspect 31) The method of embodiment 30, wherein said myeloid malignancy is acute myeloid leukemia (AML). (Aspect 32) The subject is a newly diagnosed AML patient who is ineligible for standard intensive chemotherapy. The method described in Example 31. (Aspect 33) The subject is a newly diagnosed AML patient aged 75 years or older, or a newly diagnosed and standard 33. The method of embodiment 32, wherein the AML patient has comorbidities that prevent the use of selective intensive chemotherapy. (Aspect 34) The CD70 antibody or antigen-binding fragment thereof is administered in a dose range of 0.1 to 25 mg / kg, preferably 10 mg / kg. The method according to any one of embodiments 27 to 33, wherein the dose is administered. (Aspect 35) 35. The method of any one of aspects 27 to 34, wherein the BCL-2 inhibitor is administered at a dose ranging from 100 mg to 600 mg. or the method described in paragraph 1. (Aspect 36) The combination may additionally comprise azacitidine, or the method may comprise administering azacitidine to the subject. The method of any one of embodiments 27-35, comprising the additional step of administering (Aspect 37) The azacitidine is 70 to 80 mg / m 2 , preferably 75 mg / m 2 and The method described. (Aspect 38) The combination additionally comprises decitabine, or the method further comprises administering decitabine to the subject. The method of any one of embodiments 27 to 35, comprising the additional step of administering (Aspect 39) The decitabine is 15 to 25 mg / m 2 , preferably 20 mg / m 2 The method of claim 38, wherein the Method of posting. (Aspect 40) The dose at which the CD70 antibody or antigen-binding fragment thereof is administered and the dose at which the BCL-2 inhibitor is administered 40. The method of claim 27, wherein the amounts are each selected so that the combination provides a synergistic treatment. 10. The method according to any one of the preceding claims. (Aspect 41) Any one of aspects 31 to 40, further comprising monitoring the patient's blast count. The method described in paragraph . (Aspect 42) The method of embodiment 41, wherein said patient's bone marrow blast count is reduced to less than 5%. (Aspect 43) The patient's bone marrow blast count is reduced by 5% to 25% compared to before treatment, and The method of embodiment 41, wherein the myeloblast percentage compared to is reduced by more than 50%. (Aspect 44) The method according to any one of aspects 27 to 43, which induces a partial or complete response. (Aspect 45) The method of embodiment 44, wherein the method induces a complete response accompanied by platelet recovery. (Aspect 46) 46. ​​The method of embodiment 44 or 45, which induces a complete response accompanied by neutrophil recovery. (Aspect 47) Inducing transfusion-dependence weaning of red blood cells or platelets, or both, for 8 weeks or more, in accordance with aspects 27-4. 7. The method according to any one of claims 6 to 7. (Aspect 48) The method according to any one of embodiments 27 to 47, which extends survival time. (Aspect 49) 27. Extending survival time relative to standard of care therapeutics used to treat myeloid malignancies. The method according to any one of claims 1 to 48. (Aspect 50) The method according to any one of embodiments 27 to 49, which induces a negative minimal residual disease status. (Aspect 51) The method of any one of aspects 27 to 50, further comprising providing a bone marrow transplant to the subject. (Aspect 52) administering one or more additional anti-cancer agents suitable for the treatment of malignancies, preferably myeloid malignancies; The method of any one of embodiments 27 to 51, further comprising: (Aspect 53) According to embodiment 52, the one or more additional anti-cancer agents are selected from agents suitable for treating AML. How to. (Aspect 54) The one or more additional anti-cancer agents may be a selectin inhibitor (e.g., GMI-1271); an FMS-like tyrosine kinase inhibitor (e.g., FLT3 inhibitors (e.g., midostaurin or gilteritinib); Phospho-dependent kinase inhibitors; Aminopeptidase inhibitors; JAK / STAT inhibitors; Cytarabine; Fludarabine; anthracycline compounds (e.g., daunorubicin, idarubicin); Xorubicin; Hydroxyurea; Vyxeos; IDH1 or IDH2 inhibitors, IDH Idhifa (or Enasidenib) or Tibsovo (or Ivosidenib), etc.; moothened inhibitors, glasdegib, etc.; BET bromodomain inhibitors; CD123 or CD33 targeting agents; HDAC inhibitor; LSC targeting agent; AML bone marrow niche targeting agent; NEDD8 activating enzyme inhibitor, Pebonedi Stats, etc.; G-CSF, as well as topoisomerase inhibitors, mitoxantrone, selinexon 54. The method of claim 52 or 53, wherein the compound is selected from the group consisting of cyclosporine and etoposide.

Claims

1. 1. A pharmaceutical composition for use in treating a myeloid malignancy in a human subject, comprising an antibody or antigen-binding fragment thereof that binds to CD70, the pharmaceutical composition is for administration of the antibody or antigen-binding fragment thereof in combination with a BCL-2 inhibitor and a nucleoside metabolic inhibitor; the BCL-2 inhibitor is venetoclax or a pharma- ceutical acceptable salt thereof; The subject is treated with a treatment cycle consisting of 28 days, The pharmaceutical composition, wherein the nucleoside metabolic inhibitor is for daily administration for the first 5 to 10 days of a 28 day cycle.

2. 2. The pharmaceutical composition of claim 1, wherein the nucleoside metabolic inhibitor is for administration daily for the first 7 days of the 28 day cycle.

3. 3. The pharmaceutical composition of claim 1 or 2, wherein the dose of the nucleoside metabolism inhibitor is for administration on the same day as the dose of the antibody or antigen-binding fragment thereof.

4. The pharmaceutical composition of claim 3, wherein each of the nucleoside metabolic inhibitor and the antibody or antigen-binding fragment thereof is for administration on day 1, day 2, day 3, day 4, day 5, day 6 or day 7 of the treatment cycle.

5. The pharmaceutical composition of claim 4, wherein the initial dose of the antibody or antigen-binding fragment thereof is for administration on any one of days 1 to 3 of the treatment cycle.

6. The pharmaceutical composition of claim 5, wherein the second dose of the antibody or antigen-binding fragment thereof is for administration 14 to 17 days after the first dose.

7. 7. The pharmaceutical composition of claim 6, wherein a first dose of the antibody or antigen-binding fragment thereof is for administration on day 3 and the second dose is for administration on day 17 of the 28-day cycle.

8. The pharmaceutical composition of any one of claims 1 to 6, wherein the antibody or antigen-binding fragment thereof is for administration on days 1 and 15 of the 28-day cycle.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein the pharmaceutical composition comprises the antibody or antigen-binding fragment thereof in an amount ranging from 0.1 to 25 mg / kg.

10. 10. The pharmaceutical composition of claim 9, wherein the amount of the antibody or antigen-binding fragment thereof in the pharmaceutical composition is 10 mg / kg.

11. The pharmaceutical composition of any one of claims 1 to 10, wherein the BCL-2 inhibitor is for administration in an amount ranging from 100 mg to 600 mg.

12. The antibody or antigen-binding fragment thereof comprises a variable heavy chain (VH) domain and a variable light chain (VL) domain, wherein the VH domain and the VL domain have the following CDR sequences: HCDR3 comprising or consisting of SEQ ID NO:3; HCDR2 comprising or consisting of SEQ ID NO:2; HCDR1 comprising or consisting of SEQ ID NO:1; LCDR3 comprising or consisting of SEQ ID NO:7; LCDR2 comprising or consisting of SEQ ID NO:6; and LCDR1 comprising or consisting of SEQ ID NO:5: The pharmaceutical composition according to any one of claims 1 to 11, comprising:

13. The pharmaceutical composition of claim 12, wherein the VH domain comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO:4, and the VL domain comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO:

8.

14. The pharmaceutical composition of claim 13, wherein the antibody or antigen-binding fragment thereof comprises a VH domain comprising the amino acid sequence represented by SEQ ID NO:4 and a VL domain comprising the amino acid sequence represented by SEQ ID NO:

8.

15. The pharmaceutical composition of any one of claims 1 to 14, wherein the antibody or antigen-binding fragment thereof, the BCL-2 inhibitor and the nucleoside metabolism inhibitor are formulated separately as separate compositions.

16. The pharmaceutical composition according to any one of claims 1 to 15, wherein the antibody is an IgG.

17. The pharmaceutical composition according to any one of claims 1 to 16, wherein the antibody has antibody-dependent cell-mediated cytotoxicity (ADCC) activity, complement-dependent cytotoxicity (CDC) activity, or antibody-dependent cellular phagocytosis (ADCP) activity.

18. The pharmaceutical composition of any one of claims 1 to 17, wherein the antibody comprises a defucosylated antibody domain.

19. The pharmaceutical composition of any one of claims 1 to 18, wherein the antibody is ARGX-110 (cusatuzumab).

20. The pharmaceutical composition of any one of claims 1 to 15, wherein the antigen-binding fragment is selected from the group consisting of a single chain antibody (scFv); a F(ab')2 fragment; a Fab fragment; a Fd fragment; a Fv fragment; a one-armed (monovalent) antibody; a diabody, a triabody, a tetrabody, and any antigen-binding fragment formed by a combination, assembly or conjugation of such antigen-binding fragments.

21. 21. The pharmaceutical composition of any one of claims 1 to 20, wherein the pharmaceutical composition is for use in combination with one or more additional anti-cancer agents suitable for the treatment of malignant tumors.

22. 22. The pharmaceutical composition of claim 21, wherein the malignant tumor is a myeloid malignant tumor.

23. 23. The pharmaceutical composition of claim 21 or 22, wherein the one or more additional anti-cancer agents are selected from agents suitable for the treatment of AML.

24. The one or more additional anti-cancer agents may be a selectin inhibitor; an FMS-like tyrosine kinase receptor 3 (FLT3) inhibitor; a cyclin-dependent kinase inhibitor; an aminopeptidase inhibitor; a JAK / STAT inhibitor; cytarabine; fludarabine; an anthracycline compound; doxorubicin; hydroxyurea; Vyxeos; an IDH1 or IDH2 inhibitor, such as Idhifa (or Enasidenib) or Tibsovo (or Ivosidenib); a Smoothened inhibitor, such as glasdegib; a BET bromodomain inhibitor; a CD123 or CD33 targeting agent; an HDAC inhibitor; an LSC targeting agent; an AML bone marrow niche targeting agent; a NEDD8 activating enzyme inhibitor, such as pevonedistat; 24. The pharmaceutical composition of claim 23, selected from G-CSF and topoisomerase inhibitors, such as mitoxantrone, selinexor and etoposide.

25. 25. The pharmaceutical composition of claim 24, wherein the selectin inhibitor is GMI-1271.

26. 25. The pharmaceutical composition of claim 24, wherein the FLT3 inhibitor is midostaurin or gilteritinib.

27. 25. The pharmaceutical composition of claim 24, wherein the anthracycline compound is daunorubicin or idarubicin.

28. 25. The pharmaceutical composition of claim 24, wherein the one or more additional anti-cancer agents is a CD123 targeting agent.

29. The pharmaceutical composition according to any one of claims 1 to 28, wherein the nucleoside metabolism inhibitor is a hypomethylating agent.

30. 30. The pharmaceutical composition of claim 29, wherein the hypomethylating agent is azacitidine or decitabine.

31. 31. The pharmaceutical composition of claim 30, wherein the hypomethylating agent is azacytidine.

32. The azacitidine is 70 to 80 mg / m 2 32. The pharmaceutical composition of claim 31, for administration at a dose of azacitidine of

33. The dose of azacitidine is 75 mg / m 2 33. The pharmaceutical composition of claim 32,

34. 31. The pharmaceutical composition of claim 30, wherein the hypomethylating agent is decitabine.

35. The decitabine is 15 to 25 mg / m 2 35. The pharmaceutical composition of claim 34, for administration at a dose of decitabine of

36. The dose of decitabine is 20 mg / m 2 36. The pharmaceutical composition of claim 35,

37. The pharmaceutical composition of any one of claims 1 to 36, wherein the myeloid malignancy is a newly diagnosed or relapsed / refractory myeloid malignancy.

38. The pharmaceutical composition of any one of claims 1 to 37, wherein the myeloid malignancy is selected from AML; myelodysplastic syndrome (MDS); myeloproliferative neoplasms (MPN); chronic myelogenous leukemia (CML); and chronic myelomonocytic leukemia (CMML).

39. 39. The pharmaceutical composition of claim 38, wherein the myeloid malignancy is AML.

40. The pharmaceutical composition of any one of claims 1 to 39, wherein the subject is a newly diagnosed AML patient who is ineligible for standard intensive chemotherapy.

41. 41. The pharmaceutical composition of claim 40, wherein the subject is a newly diagnosed AML patient aged 75 years or older or a newly diagnosed AML patient with comorbidities that prevent the use of standard intensive chemotherapy.

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