Combination immunotherapy and chemotherapy for the treatment of hematological malignancies

Abstract

To provide a pharmaceutical composition for Inhibiting the growth and / or proliferation of CD33-expressing cells in human subjects.SOLUTION: A pharmaceutical composition comprises four individual chemotherapeutic agents and an anti-CD33 targeting agent. The four individual chemotherapeutic agents comprise cladribine, cytarabine, mitoxantrone, and filgrastim or granulocyte colony stimulating factor. The anti-CD33 targeting agent may be, for example, HuM195 conjugated with a radionuclide such as 225Ac. It can be particularly useful for treating recurrent or refractory acute myeloid leukemia.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims the benefit of the prior U.S. Provisional Application No. 62 / 614,658, filed on 8 January 2017 under Section 119(e) of the U.S. Patent Act, which is incorporated herein by reference in its entirety.

[0002] Array List This application includes a computer-readable sequence listing incorporated herein by reference. The text file “Sequencelisting17046_ST25,” submitted via EFS in accordance with 37 CFR 1.52(e)(5) and Rule 13ter.1(a), is identical to the sequence listing that forms part of this international application.

[0003] The present invention relates to a method for treating subjects with proliferative disorders by administration of immunotherapeutic agents and chemotherapeutic agents against the CD33 epitope, and more specifically, the present invention relates to the administration of anti-CD33 antibodies and CLAG-M chemotherapeutic agents for the treatment of hematological diseases or disorders. [Background technology]

[0004] Hematological malignancies have historically been treated with high-dose chemotherapy and / or radiation. Current treatment protocols generally include combinations of chemotherapy agents such as vincristine, carmustine, cytarabine, melphalan, cyclophosphamide, daunorubicin, and steroids such as prednisone or dexamethasone. However, such treatments result in low remission rates and poor overall survival rates for hematological disorders such as acute myeloid leukemia (AML).

[0005] For example, recent advances using autologous bone marrow transplantation or peripheral blood mononuclear cell transplantation following high-dose chemotherapy have increased the complete remission rate and duration of remission in multiple myeloma (MM). However, evidence of a cure has not been obtained. The effectiveness of these available chemotherapy regimens for MM is limited because MM exhibits a low proliferation rate, acquires multidrug resistance, and has high resistance to apoptosis. Approximately 1% of all cancers and over 10% of all hematological malignancies may be attributable to MM. The majority of MM patients relapse within a few years, and in many cases, relapsed patients do not respond to salvage therapy. For more than 90% of MM patients, the disease eventually becomes chemotherapy-resistant.

[0006] Relapsed / refractory acute myeloid leukemia (RR-AML) in adults presents a particularly challenging treatment challenge. Treatment responses vary due to the use of different salvage chemotherapy regimens aimed at achieving disease remission in order to proceed to stem cell transplantation. While no universally accepted regimen has been established, various regimens such as CLAG-M (cladribine, cytarabine, mitoxantrone, and filgrastim), FLAG (fludarabine, cytarabine, idarubicin, and filgrastim), or MEC (mitoxantrone, etoposide, and cytarabine) have been used. CLAG-M has been shown to result in a 58% morphological complete response rate in prospective clinical trials for the treatment of RR-AML.

[0007] A recent retrospective study compared two commonly used regimens in RR-AML: CLAG (cladribine, cytarabine, and filgrastim) and MEC (mitoxantrone, etoposide, and cytarabine). See Price et al., Leukemia Research, vol.35, issue 3, pages 301-304. The complete response rate was 37.9% for CLAG (n=97) and 23.8% for MEC (n=65) (P=0.048), with median overall survival of 7.3 months and 4.5 months, respectively (P=0.05). In primary refractory disease, the complete response rate was 45.5% for CLAG and 22.2% for MEC (P=0.09), with median overall survival of 11 months and 4.5 months, respectively (P=0.07). In patients with relapsed AML, the complete response rate was 36.8% in CLAG and 25.9% in MEC (P=0.35), and the median overall survival was 6.7 months and 6.7 months, respectively (P=0.87). The combination of a purine nucleoside analog (cladribine) and cytarabine increases the intracellular accumulation of Ara-C-5' triphosphate (ara-C TP), which causes cytotoxicity in leukemic blasts. The addition of granulocyte colony-stimulating factor (G-CSF) further enhances the effect of the purine nucleoside analog in combination with Ara-C by activating leukemia cells and making them more receptive to chemotherapy.

[0008] Phenotypic changes that distinguish cancer cells from normal cells originating from the same tissue or cell type are often associated with one or more changes in the expression of specific gene products, including the loss of normal cell surface components or the acquisition of other components (i.e., antigens undetectable in corresponding normal non-cancerous cells). Antigens expressed by cancer cells (i.e., inside or on cancer cells) but not by normal cells, or antigens expressed by cancer cells at substantially higher levels than those found in normal cells, are often referred to as “tumor-specific antigens” or “tumor-associated antigens.” Such tumor-specific antigens can serve as markers of tumor phenotype and have been used as targets in cancer immunotherapy.

[0009] Therefore, a key strategy that can enhance the effectiveness of killing leukemic blasts when added to chemotherapy is monoclonal antibodies against markers expressed in leukemic or malignant cells. The object of the invention of this disclosure is to provide an improved method for treating proliferative disorders such as hematological diseases. [Overview of the project]

[0010] The inventions of this disclosure are based on the discovery that the administration of combinations of at least one immunotherapy agent, such as a monoclonal antibody against CD33, and combinations of two or more chemotherapeutic agents, such as a combination of cladribine, cytarabine, mitoxantrone, and granulocyte colony-stimulating factor or filgrastim (CLAG-M), has a therapeutic synergistic effect and / or improves the efficacy in treating proliferative disorders compared to the use of immunotherapy agents alone or chemotherapeutic agents (or more) alone.

[0011] Accordingly, the inventions of this disclosure relate to methods for treating subjects with proliferative disorders such as multiple myeloma, acute myeloid leukemia, myelodysplastic syndrome, or myeloproliferative neoplasms, and in particular to the treatment of relapsed or refractory states of these diseases. The methods generally involve administering an effective amount of an immunotherapy agent against CD33 and a combination of two or more effective amounts of chemotherapeutic agents. According to certain embodiments, three or more chemotherapeutic agents may be used in the disclosed methods.

[0012] The inventions of this disclosure also relate to methods for inhibiting the growth and / or proliferation of cells expressing CD33, and methods for treating diseases or disorders involving cells expressing CD33. Both methods involve targeting an effective amount of an immunotherapy agent against CD33, an effective amount of a combination of two or more chemotherapeutic agents, or an effective amount of three or more chemotherapeutic agents.

[0013] According to certain aspects of the invention of this disclosure, combinations of two or more chemotherapeutic agents, and combinations of three or more chemotherapeutic agents, may include combinations of individual chemotherapeutic agents selected from cladribine, cytarabine, mitoxantrone, and granulocyte colony-stimulating factor or filgrastim. The chemotherapeutic agents may be delivered together and / or separately during the course of treatment, according to a daily or hourly schedule specific to each chemotherapeutic agent.

[0014] According to certain aspects of the invention of this disclosure, the immunotherapy agent may include an immunotherapy agent against CD33, such as an anti-CD33 antibody conjugated to a radiolabel. The anti-CD33 antibody may be administered on one or more days from day 1 to day 30 of the treatment period. If multiple doses of the anti-CD33 antibody are administered, the doses may be the same or different.

[0015] The object of the invention of this disclosure is realized and achieved by the combination specifically outlined in the appended claims. The above-mentioned general description of the invention and the following detailed description and examples are provided to illustrate various aspects of the invention of this disclosure and should not be considered in any way as limiting any of the embodiments described. [Brief explanation of the drawing]

[0016] [Figure 1] This provides the amino acid sequence of human CD33 as shown in GenBank acceptance number NP_001763. [Modes for carrying out the invention]

[0017] The present invention relates to a method for treating proliferative disorders or disabilities by administering an effective amount of an immunotherapy agent, such as a CD33 targeting agent, and an effective amount of a chemotherapeutic agent, such as CLAG-M. Each treatment regimen (i.e., the immunotherapy agent and the chemotherapeutic agent) may be administered according to a specific dosing schedule, and the method provides the administration of each therapeutic agent sequentially (the antibody dosing schedule is completed before the chemotherapeutic dosing schedule is started, or vice versa) or simultaneously according to the dosing schedule.

[0018] definition Throughout this specification and in the appended claims, unless otherwise specified, the use of the singular form includes the plural form, and the plural form includes the singular form. For example, this specification refers to “an” antibody, “a” radionuclide, and “the” chemotherapeutic agent, but one or more of these components and / or any other components described herein may be used.

[0019] As used herein and in the claims, the words “including” and their forms do not limit the inventions of this disclosure to exclude any modifications or additions. Furthermore, although the inventions of this disclosure are described in terms of “including,” the processes, materials, and compositions detailed herein may be described as essentially or as being. For example, certain aspects of the invention are described relating to a method comprising administering an immunotherapy agent for CD33 and a chemotherapy regimen such as CLAG-M, but methods “essentially consisting of” or “consisting of” the administration of an immunotherapy agent for CD33 and a chemotherapy regimen are also within the scope of the invention. In this context, “essentially consisting of” means that any additional components do not substantially affect the effectiveness of the method.

[0020] Furthermore, beyond the examples provided, or where otherwise indicated, all figures representing quantities of components used herein should be understood in all cases to be modified by the term “approximately.” Therefore, unless otherwise indicated, the numerical parameters described herein are approximations that may vary depending on the desired properties obtained by the invention of this disclosure. At a minimum, and not as an attempt to limit the application of the principle of equivalents to the claims, each numerical parameter should be interpreted at least in light of the number of significant digits reported and by applying common rounding techniques. Thus, the term “approximately,” when used before numerical expressions such as temperature, time, quantity, and concentration (including ranges), indicates an approximation that may vary by ±10%, ±5%, or ±1%.

[0021] As used herein, the term “administer” with respect to targeted agents such as antibodies, antibody fragments, Fab fragments, or aptamers means of delivering the agent to the body of the target by any known method suitable for antibody delivery. Specific modes of administration include, but are not limited to, intravenous, transdermal, subcutaneous, intraperitoneal, subarachnoid, and intratumoral administration. Exemplary methods of antibody administration may be substantially described in International Publication WO2016 / 187514, incorporated herein by reference. For example, according to certain embodiments, the targeted agent may be administered as a patient-specific therapeutic composition contained in a single-dose container, the total volume of which may be administered to the patient in a single therapeutic session. The composition may include a monoclonal antibody or antibody fragment and a pharmaceutically acceptable carrier, and the dose of the effector molecule (e.g., radionuclide) of the monoclonal antibody and the total protein amount of the monoclonal antibody may depend on at least one patient-specific parameter. Patient-specific parameters include, but are not limited to, the patient’s weight, age, height, sex, condition, and medical history.

[0022] Furthermore, in the present invention, antibodies or antibody fragments can be formulated using one or more routinely used pharmaceutically acceptable carriers. Such carriers are well known to those skilled in the art. For example, injectable drug delivery systems include solutions, suspensions, gels, microspheres, and polymeric injectables, and may include excipients such as solubility modifiers (e.g., ethanol, propylene glycol, and sucrose) and polymers (e.g., polycaprylactone and PLGA). Exemplary formulations may be substantially as described in International Publication WO2017 / 155937, which is incorporated herein by reference. For example, according to a particular embodiment, a formulation may contain 0.5% to 5.0% (w / v) of an excipient selected from the group consisting of ascorbic acid, polyvinylpyrrolidone (PVP), human serum albumin (HSA), water-soluble salts of HSA, and mixtures thereof. Certain formulations may contain 0.5–5% ascorbic acid, 0.5–4% polyvinylpyrrolidone (PVP), and a monoclonal antibody in 50 mm PBS buffer (Ph7).

[0023] As used herein, the term “antibody” includes, but is not limited to, (a) immunoglobulin molecules comprising two heavy chains and two light chains that recognize an antigen, (b) polyclonal and monoclonal immunoglobulin molecules, (c) monovalent and bivalent fragments thereof (e.g., di-Fab), and (d) bispecific forms thereof. Immunoglobulin molecules may originate from any of the commonly known classes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG subclasses are also well known to those skilled in the art and include, but are not limited to, human IgG1, IgG2, IgG3, and IgG4. Antibodies can be of both natural and non-natural origin (e.g., IgG-Fc silent). Furthermore, antibodies include chimeric antibodies, fully synthetic antibodies, single-chain antibodies, and fragments thereof. Antibodies may be human, humanized, or non-human.

[0024] As used herein, “immunoreactivity” refers to a measure of the ability of an immunoglobulin to recognize and bind to a specific antigen. “Specific binding,” “specifically binding,” or “binding” refers to an antibody that binds to an antigen or an epitope within an antigen with a higher affinity than to other antigens. Typically, an antibody has an affinity of approximately 1 × 10⁻⁶. -8 M or less, for example, about 1 × 10 -9 M or less, approximately 1×10 -10 M or less, approximately 1×10 -11 M or less, or approximately 1 × 10 -12 Equilibrium dissociation constants (K) less than or equal to M D Typically, the K1 is at least 100 times less than its KD for binding to nonspecific antigens (e.g., BSA, casein). D The antibody then binds to the antigen or an epitope within the antigen. The dissociation constant may be measured using standard procedures. However, antibodies that specifically bind to an antigen or an epitope within an antigen may cross-react to the same antigen (homologous) from other related antigens, such as humans or monkeys, e.g., Macaca fascicularis (cynomolgus monkey, cyno), Pan troglodytes (chimpanzee, chimp), or Callitrisrix jacchus (common marmoset, marmoset).

[0025] As used herein, “anti-CD33 targeting agent” is an antibody, antibody fragment, peptide, Fab fragment, or aptamer that binds to any available epitope of CD33. In a particular embodiment, the anti-CD33 targeting agent is a humanized antibody against CD33, such as lintuzumab (HuM195), gemtuzumab, or vadastuximab. In a particular embodiment, the anti-CD33 targeting agent binds to an epitope recognized by the monoclonal antibody “lintuzumab” or “HuM195”. HuM195 is known, and similarly, methods for producing it are also known.

[0026] An "epitope" refers to a target molecular site (e.g., at least a portion of an antigen) that is recognized and can be bound by a targeting agent such as an antibody, antibody fragment, Fab fragment, or aptamer. In the case of protein antigens, for example, this may refer to a region of the protein to which the antibody binds (i.e., amino acids, and especially their side chains). Overlapping epitopes contain at least 1 to 5 common amino acid residues. Methods for identifying antibody epitopes are known to those skilled in the art, including, for example, those described in Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988).

[0027] As used herein, “cancer” includes, but is not limited to, solid tumors (e.g., tumors) and hematological malignancies.

[0028] "Hematological disorders" or "hematological conditions" can be understood to refer to at least blood cancers. Such cancers originate from hematopoietic tissues such as bone marrow or other cells of the immune system. Hematological disorders or hematological conditions include, but are not limited to, leukemia (e.g., acute myeloid leukemia (AML), acute promyelocytic leukemia, acute lymphoblastic leukemia (ALL), acute mixed lineage leukemia, chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), hairy cell leukemia, and macrogranular lymphocytic leukemia), myelodysplastic syndromes (MDS), and myeloproliferative disorders (polycythemia vera, essential thrombocytosis). This includes disease, primary myelofibrosis and chronic myeloid leukemia, lymphoma, multiple myeloma, MGUS and similar disorders, Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), primary mediastinal large B-cell lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, transformed follicular lymphoma, perisplenic zone lymphoma, lymphocytic lymphoma, T-cell lymphoma, and other B-cell malignancies.

[0029] "Solid tumors" include, but are not limited to, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, malignant melanoma of the skin or eyeball, uterine cancer, ovarian cancer, prostate cancer, rectal cancer, anal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, pediatric tumors, bladder cancer, kidney or ureter cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axial tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermal carcinoma, squamous cell carcinoma, and environmentally induced cancers, including those induced by asbestos.

[0030] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" is a mechanism for inducing cell death that relies on the interaction between antibody-coated target cells and effector cells with lytic activity, such as natural killer (NK) cells, monocytes, macrophages, and neutrophils, via the Fc gamma receptor (FcγR) expressed on effector cells. For example, NK cells express FcγRIIIa, while monocytes express FcγRI, FcγRII, and FcvRIIIa. The death of antibody-coated target cells, such as CD38-expressing cells, occurs as a result of effector cell activity mediated by the secretion of pore-forming proteins and proteases.

[0031] Complement-dependent cell-mediated cytotoxicity (ADCC) refers to a mechanism for inducing cell death in which the Fc effector domain of a target-binding antibody binds to and activates complement component C1q, which then activates the complement cascade, leading to the death of target cells. Complement activation may also result in the accumulation of complement components on the surface of target cells, which promote ADCC by binding to complement receptors (e.g., CR3) on leukocytes.

[0032] "Apoptosis" refers to a programmed cell death mechanism in which antibody binding to target cells disrupts essential cellular signaling pathways, leading to the self-destruction of the cells.

[0033] In certain embodiments, anti-CD33 targeting agents can be labeled with radioisotopes.131 I or 225Methods for labeling proteins such as antibodies with radioisotopes such as Ac are known. These methods are described, for example, in International Publication No. WO2017 / 155937 or International Application No. PCT / US18 / 44531. For example, according to a particular embodiment, an anti-CD33 targeting agent may be labeled by (a) reacting the targeting agent with a chelating agent in a buffer, (b) reacting the chelated targeting agent with a radionuclide in a buffer, (c) stopping the reaction by adding a quenching chelating agent (e.g., diethylenetriaminepentaacetic acid (DTPA)), and (d) purifying the radiolabeled chelated targeting agent. Exemplary chelating agents include compounds having a dual functionality, having the ability to covalently bind to biological carriers such as antibodies in addition to sequestering metal ions.Examples of chelating agents include, but are not limited to, S-2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecanetetraacetic acid (p-SCN-Bn-DOTA), diethylenetriaminepentaacetic acid (DTPA); ethylenediaminetetraacetic acid (EDTA); 1,4,7,10-tetra-azacyclododecane-N,N′,N″,N′″-tetraacetic acid (DOTA); p-isothiocyanatobenzyl-1,4,7,10-tetra-azacyclododecane-1,4,7,10-tetraacetic acid (p- SCN-Bz-DOTA); 1,4,7,10-Tetra-azacyclododecane-N,N′,N″-triacetic acid (DO3A); 1,4,7,10-Tetra-azacyclododecane-1,4,7,10-Tetrakis(2-propionic acid) (DOTMA); 3,6,9-Triaza-12-oxa-3,6,9-Tricarboxymethylene-10-carboxy-13-phenyl-tridecanoic acid ("B-19036"); 1,4,7-Triazacyclononane-N,N′,N″-triacetic acid (NOTA); 1, 4,8,11-Tetra-azacyclotetradecane-N,N′,N″,N′″-tetraacetic acid (TETA); triethylenetetraaminehexaacetic acid (TTHA); trans-1,2-diaminohexanetetraacetic acid (CYDTA); 1,4,7,10-Tetra-azacyclododecane-1-(2-hydroxypropyl)-4,7,10-triacetic acid (HP-DO3A); trans-cyclohexane-diaminetetraacetic acid (CDTA); trans-(1,2)-cyclohexanediethylenetriaminepentaacetic acid (CDTP A) This includes compounds such as 1-oxa-4,7,10-triazacyclododecane-N,N′,N″-triacetic acid (OTTA); 1,4,7,10-tetra-azacyclododecane-1,4,7,10-tetrakis{3-(4-carboxyl)-butanoic acid}; 1,4,7,10-tetra-azacyclododecane-1,4,7,10-tetrakis(acetic acid-methylamide); 1,4,7,10-tetra-azacyclododecane-1,4,7,10-tetrakis(methylenephosphonic acid); and their derivatives.

[0034] As used herein, “radioisotopes” and “radionuclides” may be used interchangeably and may refer to alpha-emitting isotopes, beta-emitting isotopes, and / or gamma-emitting isotopes. Examples of radioisotopes include: 131 I, 125 I, 123 I, 90 Y, 177 Lu, 186 Re, 188 Re, 89 Sr, 153 Sm, 32 P, 225 Ac, 213 Bi, 213 Po, 211 At, 212 Bi, 213 Bi, 223 Ra, 227 Th, 149 Tb, 137 Cs, 212 Pb and 103 Pd.

[0035] According to a particular embodiment, an anti-CD33 targeting agent is 131 It may also be an antibody radiolabeled with I (" 131 (labeled as "I"), the effective dose may be, for example, less than 1200 mCi (i.e., administered to the subject) 131 The amount of I delivers a whole-body radiation dose of less than 1200 mCi). In certain embodiments, the antibody 131 When labeled with I, the effective amount may be less than 1000 mCi, less than 750 mCi, less than 500 mCi, less than 250 mCi, less than 200 mCi, less than 150 mCi, less than 100 mCi, less than 50 mCi, less than 40 mCi, less than 30 mCi, less than 20 mCi, or less than 10 mCi.

[0036] According to a particular aspect, 131The effective dose of I-labeled antibody is 10 mCi to 200 mCi. Examples of effective doses, but not limited to, include 50 mCi to 100 mCi, 50 mCi to 150 mCi, 50 mCi to 200 mCi, 60 mCi to 140 mCi, 70 mCi to 130 mCi, 80 mCi to 120 mCi, 90 mCi to 110 mCi, 100 mCi to 150 mCi, 50 mCi, 60 mCi, 70 mCi, 80 mCi, 90 mCi, 100 mCi, 110 mCi, 120 mCi, 130 mCi, 140 mCi, 150 mCi, or 200 mCi.

[0037] According to a particular aspect, 131 The effective dose of I-labeled antibody is 200 mCi to 1200 mCi. Examples of effective doses, though not limited to these, include 200 mCi to 300 mCi, 200 mCi to 400 mCi, 200 mCi to 500 mCi, 200 mCi to 600 mCi, 200 mCi to 700 mCi, 200 mCi to 800 mCi, 200 mCi to 900 mCi, 200 mCi to 1000 mCi, 200 mCi to 1100 mCi, 300 mCi to 1200 mCi, 400 mCi to 1200 mCi, and 500 mCi to 1 Examples include 200mCi, 600mCi-1200mCi, 700mCi-1200mCi, 800mCi-1200mCi, 900mCi-1200mCi, 1000mCi-1200mCi, 50mCi, 100mCi, 150mCi, 200mCi, 300mCi, 400mCi, 500mCi, 600mCi, 700mCi, 800mCi, 900mCi, 1000mCi, or 1100mCi.

[0038] According to a particular embodiment, an anti-CD33 targeting agent is 225 The antibody may also be radiolabeled with Ac (" 225 (As labeled with Ac), the effective dose may be, for example, less than 5.0 μCi / kg (i.e., administered to the subject) 225 The amount of Ac administered delivers a radiation dose of less than 5.0 μCi per kilogram of the subject's body weight. According to certain embodiments, the antibody 225When labeled with AC, the effective dose is 4.5 μCi / kg, 4.0 μCi / kg, 3.5 μCi / kg, 3.0 μCi / kg, 2.5 μCi / kg, 2.0 μCi / kg, 1.5 μCi / kg, 1.0 μCi / kg, 0.9 μCi / kg, 0.8 μCi / kg, 0.7 μCi / kg, 0.6 μCi / kg, 0.5 μCi / kg, 0.4 μCi / kg, 0.3 μCi / kg, 0.2 μCi / kg, 0.1 μCi / kg, or less than 0.05 μCi / kg.

[0039] According to certain aspects, antibodies 225 When labeled with Ac, the effective amount is 0.05μCi / kg~0.1μCi / kg, 0.1μCi / kg~0.2μCi / kg, 0.2μCi / kg~0.3μCi / kg, 0.3μCi / kg~0.4μCi / kg , 0.4μCi / kg~0.5μCi / kg, 0.5μCi / kg~0.6μCi / kg, 0.6μCi / kg~0.7μCi / kg, 0.7μCi / kg~0.8μCi / kg, 0.8μCi / kg~0.9μ Ci / kg, 0.9μCi / kg~1.0μCi / kg, 1.0μCi / kg~1.5μCi / kg, 1.5μCi / kg~2.0μCi / kg, 2.0μCi / kg~2.5μCi / kg, 2.5μCi / kg ~3.0μCi / kg, 3.0μCi / kg~3.5μCi / kg, 3.5μCi / kg~4.0μCi / kg, 4.0μCi / kg~4.5μCi / kg, or 4.5μCi / kg~5.0μCi / kg.

[0040] According to certain aspects, antibodies 225 When labeled with Ac, the effective amounts are 0.05μCi / kg, 0.1μCi / kg, 0.2μCi / kg, 0.3μCi / kg, 0.4μCi / kg, 0.5μCi / kg, 0.6μCi / kg, 0.7μCi / kg, 0.8μ Ci / kg, 0.9μCi / kg, 1.0μCi / kg, 1.5μCi / kg, 2.0μCi / kg, 2.5μCi / kg, 3.0μCi / kg, 3.5μCi / kg, 4.0μCi / kg or 4.5μCi / kg.

[0041] According to certain aspects of the invention of this disclosure, when the anti-CD33 targeting agent is labeled with a radioisotope, the majority of the targeting agent (antibody, antibody fragment, etc.) administered to the target typically consists of unlabeled targeting agents, with only a small number being labeled targeting agents. The ratio of labeled targeting agents to unlabeled targeting agents can be adjusted using known methods. Accordingly, according to certain aspects of the invention of this disclosure, the anti-CD33 targeting agent may be provided in a total protein amount of up to 100 mg, for example, up to 60 mg, for example, 5 mg to 45 mg, or 0.01 mg / kg of patient body weight to 16.0 mg / kg of patient body weight, for example, 0.01 mg / kg to 10.0 mg / kg, or 0.05 mg / kg to 5.0 mg / kg, or 0.01 mg / kg to 1.0 mg / kg, or 0.01 mg / kg to 0.6 mg / kg of patient body weight, or 0.01 mg / kg, 0.015 mg / kg of patient body weight, 0.02 mg / kg of patient body weight, or 0.04 mg / kg of patient body weight, or 0.06 mg / kg of patient body weight, for a total protein amount. According to certain aspects of the invention of this disclosure, the effective dose of the anti-CD33 antibody is 10 mg / m² 2 Less than, for example, about 6 mg / m² 2 , or 3 mg / m² 2 , or even 2 mg / m² 2 This could be the dosage.

[0042] According to certain aspects of the invention of this disclosure, when the anti-CD33 targeting agent is radiolabeled, the radiolabeled anti-CD33 targeting agent may comprise a labeled fraction and an unlabeled fraction, where the ratio of labeled fraction to unlabeled fraction may be about 0.01:10 to 1:1, for example, the ratio of labeled fraction to unlabeled fraction may be 0.1:10 to 1:1. Furthermore, the radiolabeled anti-CD33 targeting agent may be provided as a single-dose composition tailored to a specific patient, and the amounts of labeled and unlabeled anti-CD33 targeting agent in the composition may depend at least on the patient's weight, age, and / or medical condition, as detailed in International Publication WO2016 / 187514.

[0043] As used herein, the term “subject” includes, but is not limited to, humans, non-human primates, dogs, cats, horses, sheep, goats, cattle, rabbits, pigs, rats, and mice. If the subject is human, the subject may be of any age. For example, a subject may be 60 years or older, 65 years or older, 70 years or older, 75 years or older, 80 years or older, 85 years or older, or 90 years or older. Alternatively, a subject may be 50 years or younger, 45 years or younger, 40 years or younger, 35 years or younger, 30 years or younger, 25 years or younger, or 20 years or younger. In the case of a human subject with cancer, the subject may be newly diagnosed, or may have relapsed and / or refractory cancer, or may be in remission.

[0044] As used herein, “treating” a subject with cancer includes, but is not limited to, (i) slowing, stopping or reversing the progression of cancer; (ii) slowing, stopping or reversing the progression of cancer symptoms; (iii) reducing the likelihood of cancer recurrence; and / or (iv) reducing the likelihood of cancer symptoms recurring. In certain preferred embodiments, treating a subject with cancer means (i) reversing the progression of cancer until, ideally, the cancer is eliminated; and / or (ii) reversing the progression of cancer symptoms until, ideally, the symptoms are eliminated; and / or (iii) reducing or eliminating the likelihood of recurrence (i.e., consolidation therapy that ideally destroys any remaining cancer cells).

[0045] In the context of this invention, “chemotherapeutic agent” means a compound that inhibits or kills proliferating cells and can be used in the treatment of cancer or is approved for use in the treatment of cancer. Exemplary chemotherapeutic agents include alkylating agents, plant alkaloids, antimetabolites, antitumor antibiotics, topoisomerase inhibitors, mitotic inhibitors, corticosteroids, and cell division inhibitors that prevent, inhibit, interfere with, or delay cell division at the mitotic or cytokinesis level.

[0046] The "therapeutic dose" or "effective dose" refers to the amount that is effective in the required dosage and for the required duration to achieve the desired therapeutic outcome. The therapeutic dose may vary depending on factors such as the individual's condition, age, sex, and weight, as well as the ability of the therapeutic agent or combination of therapeutic agents to elicit the desired response in the individual. Examples of indicators of an effective therapeutic agent or combination of therapeutic agents include, for example, improved patient health, reduced tumor burden, cessation or slowing of tumor growth, and / or the absence of metastasis of cancer cells to other parts of the body. In certain embodiments, the "therapeutic dose" or "effective dose" refers to the amount of anti-CD33 targeting agent that can deplete or reduce the total number of CD33-expressing cells, or inhibit the growth of CD33-expressing cells.

[0047] As used herein, “depleting” with respect to CD33-expressing cells means reducing a population of at least one type of cell that expresses or overexpresses CD33 (for example, at least one type of peripheral blood lymphocytes of the subject or at least one type of bone marrow lymphocytes of the subject). According to certain aspects of the present invention, lymphopenia of the subject is determined by measuring the level of peripheral blood lymphocytes of the subject. For example, the lymphocyte population of the subject is depleted if at least one population of peripheral blood lymphocytes of the subject is reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 99%, etc.

[0048] "Growth inhibition" means a measurable reduction or delay in the growth of malignant cells or tissues (e.g., tumors) in vitro or in vivo when exposed to a therapeutic agent or combination of drugs, compared to a reduction or delay in the growth of the same cells or tissues in the absence of the therapeutic agent or combination of drugs. Inhibition of malignant cell or tissue growth in vitro or in vivo may be at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0049] As used herein, “synergistic combination” is a combination of monotherapies that can provide therapeutic effects comparable to the efficacy of monotherapy while mitigating adverse side effects, such as damage to non-target tissues, immune status, and other clinical signs. Alternatively, synergistic combinations can provide improved efficacy, which may be measured by total tumor cell count, time to recurrence, and other indicators of the patient’s health.

[0050] The synergistic combination of the inventions disclosed herein involves combining an immunotherapy agent against CD33, i.e., an agent that targets the suppression or blocking of the function of CD33, such as a monoclonal antibody against CD33, with an agent that targets the reduction of the proliferation of a specific cell type, such as a chemotherapeutic agent or a combination of chemotherapeutic agents.

[0051] Throughout this application, various publications are cited. The disclosures of these publications are incorporated herein by reference to this application in order to more fully illustrate the prior art to which the present invention relates.

[0052] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art in which the inventions of this disclosure pertain. Similar or equivalent methods and materials may be used in the practices or tests described herein, but suitable methods and materials are described below.

[0053] Embodiments of the present invention CD33 overexpression is common in many hematological malignancies, including AML, CML, and MDS. In AML, 85–90% of patients express CD33, which has led to the development of targeted therapies such as gemtuzumab ozogamicin (Mylotarg). Approximately 96% of MDS patients express CD33 in myeloblasts (Sanford et al., “CD33 is frequently expressed in cases of myelodysplastic syndrome and chronic myelomonocytic leukemia with elevated blast count,” 2016, Leukemia & Lymphoma, vol.57(8):1965-1968). In another study, MDS patients showed approximately twice as many CD33 molecules per bone marrow cell compared to control samples (Jilani, et al., “Differences in CD33 intensity between various myeloid neoplasms,” 2002, Am J Clin Pathol 2002, vol.118:560-566). The CD33 antigen is expressed in virtually all cases of CML. Furthermore, the prognosis is poor for patients over 60 years of age, with only 10% to 15% of AML patients achieving a 4-year disease-free survival period. This high relapse rate in AML patients and the poor prognosis in older patients are linked to CD33. + This emphasizes the need for novel therapeutics that preferentially target cells.

[0054] Accordingly, the method disclosed herein includes the administration of an immunotherapy agent against CD33. The method may be used to treat proliferative disorders such as hematological diseases or disorders, and / or to inhibit the growth and / or proliferation of cells expressing CD33, and / or to treat diseases or disorders involving cells expressing or overexpressing CD33. Furthermore, the method can treat relapsed / refractory hematological diseases or disorders, which are selected from multiple myeloma, acute myeloid leukemia, myelodysplastic syndromes, and myeloproliferative neoplasms.

[0055] Human CD33 has the amino acid sequence shown in GenBank acceptance number NP_001763 and Sequence ID No. 1 (Figure 1). CD33 is a 67 kD type I transmembrane receptor glycoprotein that can function as a sialic acid-dependent cell adhesion molecule. CD33 has a long N-terminal extracellular domain, a helical transmembrane domain, and a short C-terminal cytoplasmic domain. CD33 is expressed on early myeloid progenitor cells and myeloid leukemia (e.g., acute myeloid leukemia, AML) cells, but not on stem cells.

[0056] Referring to Figure 1, amino acid residues 1-259 represent the extracellular domain, amino acids 260-282 represent the helical transmembrane domain, and amino acids 283-364 represent the cytoplasmic domain (intracellular). The extracellular domain of CD33 (i.e., W22R, R69G, S128N) contains at least three known single nucleotide polymorphisms ("SNPs"). Therefore, the extracellular domain of human (Homo sapiens) CD33 may have the amino acid sequence of Sequence ID No. 1 containing one or more of these SNPs.

[0057] Recent studies suggest a role for CD33 in regulating inflammatory and immune responses through attenuation of tyrosine kinase-driven signaling pathways. For example, in vitro studies have demonstrated that CD33 constitutively suppresses the production of inflammatory cytokines such as IL-1β, TNF-α, and IL-8 by human monocytes in sialic acid ligand-dependent and SOCS3-dependent manner. Conversely, a decrease in cell surface CD33 or disruption of sialic acid binding may increase the activity of p38 mitogen-activated protein kinase (MAPK), enhancing cytokine secretion and cytokine-induced cell proliferation.

[0058] Antibodies against CD33, such as lintuzumab (HuM195), gemtuzumab, and vadastuximab, have been and continue to be clinically evaluated for their efficacy in treating hematological malignancies, including acute myeloid leukemia (AML), and plasma cell disorders. Each antibody is known to bind to different parts of the extracellular domain of CD33, and each exhibits a different clinical response (e.g., antitumor effect). Gemtuzumab is available from Pfizer as Mylotarg®, and vadastuximab is available from Seattle Genetics as vadastuximab talirin.

[0059] For example, the antibody lintuzumab (HuM195) demonstrated anti-leukemic efficacy in the treatment of AML. HuM195 is a recombinant humanized anti-CD33 monoclonal antibody originally manufactured by Protein Design Labs, Inc. (Fremont, California). M195 is a monoclonal IgG2a antibody that binds to CD33. M195 is derived from mice immunized with live human leukemic myeloblasts. HuM195 was constructed by transplanting the complementarity-determining region of M195 into a human IgG1 framework and scaffold. HuM195 induced antibody-dependent cell-mediated cytotoxicity using human peripheral blood mononuclear cells as effectors. Four clinical trials investigated only native (i.e., non-conjugated) HuM195 in patients with relapsed or refractory AML and CML. Fever, chills, and nausea were the most common toxicities. No human anti-human antibody responses were observed. Some patients showed beneficial biological activity in terms of a reduction in myeloblast cells. Since those who benefited most had fewer blast cells at the start of treatment, it is suggested that Hum195 may be more effective in treating minimal residual disease or cytoreduced diseases.

[0060] The proposed methods for these antibodies to eliminate CD33-positive cells include antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cell-mediated cytotoxicity (CDC), and apoptosis.

[0061] To evaluate the ADCC activity of antibodies that bind to specific antibodies, such as antibodies against CD33, the antibodies may be added to antigen-expressing cells in combination with immunoeffector cells, and each can be activated by an antigen-antibody complex that induces cell lysis of the antigen-expressing cells. Cell lysis is usually detected by the release of a label (e.g., a radioactive substrate, a fluorescent dye, or a native intracellular protein) from the lysed cells. Exemplary effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and NK cells.

[0062] For example, in an exemplary assay of the ADCC activity of an anti-CD33 antibody, CD33-expressing cells 51 The cells may be labeled with Cr and thoroughly washed. Anti-CD33 antibody may be added to CD33-expressing cells at various concentrations, and the assay is initiated by adding effector cells (e.g., NK cells from peripheral blood mononuclear cells). After incubation at 37°C at various time intervals, the assay is stopped by centrifugation, and the lysed cells are extracted. 51 Cr release is measured using a scintillation counter. The cytotoxicity rate can be calculated as the maximum solubility percentage that can be induced by adding 3% perchlorate to CD33-expressing cells.

[0063] In an exemplary cytotoxic assay, tetrazolium salts may be added to CD33-expressing cells treated with varying amounts of anti-CD33 antibody. In living mitochondria, XTT is reduced to an orange product by mitochondrial dehydrogenase and transferred to the cell surface. The orange product can be optically quantified and reflects the number of living cells. Alternatively, esterases from living cells are known to hydrolyze colorless calcein as a fluorescent molecule. The fluorescence can be measured and quantified and reflects the number of living cells in the sample. The total number of dead cells may be measured using propidium iodide, which is excluded from living cells by an intact membrane. Fluorescence from propidium iodide in dead cells can be quantified by flow cytometry.

[0064] To evaluate CDC, it may be necessary to include complement proteins in cytotoxic assays. Measuring apoptosis induction does not require the addition of NK cells or complement proteins in cytotoxic assays.

[0065] As mentioned above, the treatment of certain cancers with monoclonal antibodies against CD33 has yielded various successes. In initial studies using a non-conjugate mouse anti-CD33 antibody (M195), a small number of patients showed a transient decrease in peripheral blast counts at saturated or supersaturated doses. Subsequent studies using lintuzumab (HuM195), a humanized version of M195 with an 8-fold higher binding affinity, showed antibody-dependent cell-mediated cytotoxicity (ADCC), unlike M195. Furthermore, while limited studies showed some activity in acute promyelocytic leukemia (APL) when used in patients with minimal residual disease, lintuzumab had very little activity as a monotherapy in AML, even at supersaturated doses, and achieving complete or partial remission, limited to patients with low tumor burden and complete blocking of the CD33 binding site throughout 4 weeks, was rare. As suggested by a small clinical trial in which very high doses of lintuzumab were administered weekly for 5 weeks, followed by bi-weekly administration to patients who showed clinical benefit, repeated administration of ultrasaturated doses may potentially increase efficacy.

[0066] One approach to improving the efficacy of immunotherapeutic agents (i.e., anti-CD33 antibodies) involves the use of multispecific antibodies. Accordingly, according to certain aspects of the invention of this disclosure, the method may include the administration of an immunotherapeutic agent, the immunotherapeutic agent may include a multispecific antibody against a first epitope of CD33 and a second epitope of CD33, or against the epitope of CD33 and the epitopes of one or more additional different antigens. Accordingly, the immunotherapeutic agent may include a multispecific antibody comprising at least a first target recognition component that specifically binds to the epitope of CD33 and a second target recognition component that specifically binds to the epitope of an antigen other than CD33.

[0067] Additional different antigens may be antigens differentially expressed on cells involved in hematological disorders or impairments, and / or cells involved in solid tumors. For example, additional different antigens may be mesothelin, TSHR, CD19, CD123, CD22, CD30, CD45, CD171, CD138, CS-1, CLL-1, GD2, GD3, B-cell maturation antigen (BCMA), Tn Ag, Prostatic membrane antigen (PSMA), ROR1, FLT3, FAP, TAG72, CD38, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, Interleukin-11 receptor α (IL-11Rα), PSCA, PRSS21, VEGFR2, LewisY, CD24, Platelet-derived growth factor-β (PDGFR-β), SSEA-4, CD20, Folate receptor α (FRa), ERBB2 (Her2 / neu), MUCl, Epidermal growth factor receptor (EGFR), EGFRvIII, NCAM, Prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gplOO, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, folate receptor β, TEM1 / CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-la, MAGE-A1, Legumain, HPV E6,E7, MAGE Al, MAGEA3, MAGEA3 / A6, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, Prostein, survivor and telomerase, PCTA-I / galectin 8, KRAS, MelanA / MARTI, Ras variant, hTERT, sarcoma translocation breakpoint, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin B1, MYCN, RhoC, TRP-2, CYP1B 1, BORIS, SART3, PAX5, OY-TES 1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxylesterase, mut The following can be selected: hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, GPA7, and IGLL1.

[0068] The first target recognition component may comprise one of the first full-length heavy chain and the first full-length light chain, the first Fab fragment, or the first single-chain variable fragment (scFv). Furthermore, the first target recognition component may be derived from lintuzumab (HuM195), gemtuzumab, or vadasutuximab. The second target recognition component may comprise one of the second full-length heavy chain and the second full-length light chain, the second Fab fragment, or the second single-chain variable fragment (scFv). Furthermore, the second target recognition component may be derived from any of the additional different antigens listed above.

[0069] Multispecific antibodies may be recombinant antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, or antibody fragments.

[0070] Alternatively, the present invention envisions a method comprising administering a first antibody against at least one antigen of CD33, and a second antibody, wherein the second antibody is against a different epitope of CD33 than the first antibody, or against an epitope of a different antigen, such as an antigen selected from the list presented above.

[0071] Another approach to improving the therapeutic effect of the monoclonal antibody used in the method of the present invention involves labeling the antibody with a radionuclide. In the treatment of a patient with a radionuclide-labeled monoclonal antibody against CD33, the radionuclide can be localized to cells expressing CD33 at tumor sites, etc., thereby killing those tumor cells. Accordingly, the present invention relates to a method for treating proliferative disorders, comprising administering an effective amount of a monoclonal antibody against CD33 and an effective amount of a chemotherapy regimen, wherein the antibody is labeled with an effector molecule such as a radionuclide. According to a particular aspect of the present invention, the monoclonal antibody against CD33 may be lintuzumab (HuM195), gemtuzumab, and / or vadasutuximab.

[0072] According to a particular aspect of the present invention, the radiolabeled anti-CD33 targeting agent is radiolabeled HuM195 (lintuzumab). The radiolabeled antibodies envisioned in the present invention are not limited to, 131 I-HuM195, 125 I-HuM195, 123 I-HuM195, 90 Y-HuM195, 177 Lu-HuM195, 186 Re-HuM195, 188 Re-HuM195, 89 Sr-HuM195, 153 Sm-HuM195, 32 P-HuM195, 225 AcHuM195, 213 Bi-HuM195, 213 Po-HuM195, 211 At-HuM195, 212 Bi-HuM195, 213 Bi-HuM195, 223 Ra-HuM195, 227 Th-HuM195, 149 Tb-HuM195, 131 I-HuM195, 137 Cs-HuM195, 212 Pb-HuM195 and 103Contains Pd-HuM195. Radiolabeled anti-CD33 antibody, 131 I-HuM195 or 225 This is the Ac-HuM195.

[0073] According to a particular aspect of the invention disclosed herein, the targeting agent is 131 I or 225 It may be labeled with Ac and may be at least 5 times more effective than a control monoclonal antibody in inducing cell death of lymphoblasts or myeloma cells, and the control targeting agent is 131 I or 225 This includes an unlabeled targeting agent for the same antigenic epitope as an Ac-labeled targeting agent. For example, 131 I or 225 Ac-labeled monoclonal antibodies may be at least 10-fold, at least 20-fold, at least 50-fold, or at least 100-fold more effective than control monoclonal antibodies in inducing cell death in lymphoblasts or myeloma cells.

[0074] According to certain aspects of the invention of this disclosure, the method may involve the administration of labeled and unlabeled (e.g., "naked") fractions of an anti-CD33 targeting agent, such as an antibody or antibody fragment. For example, the unlabeled fraction may contain the same antibody against the same epitope as the labeled fraction. In this way, the total radioactivity of the antibody can be varied or kept constant, while the overall antibody protein concentration can be kept constant or varied. For example, the total protein concentration of the administered unlabeled antibody fraction may vary depending on the exact nature of the disease being treated, the patient's age and weight, the identity of the monoclonal antibody, and the label (e.g., radionuclide) selected for labeling the monoclonal antibody.

[0075] According to a particular aspect of the invention disclosed herein, the anti-CD33 targeting agent is administered at a radiation dose of 10 mCi to 1,200 mCi, as already described in detail. 131 It may be I-HuM195. According to a particular aspect of the present invention, the anti-CD33 targeting agent is 131The anti-CD33 antibody labeled with I may be administered in one or more doses according to any of the dosing schedules listed herein until the subject receives a cumulative radiation dose of 10 mCi to 300 mCi, such as 20 mCi to 200 mCi or 20 mCi to 100 mCi.

[0076] According to a particular aspect of the invention disclosed herein, the anti-CD33 targeting agent is administered at a radiation dose of less than 5.0 μCi / kg, as already described in detail. 225 It may be AcHuM195 (i.e., administered to the subject) 225 Antibodies labeled with Ac deliver a radiation dose of less than 5.0 μCi per kg of body weight of the subject. According to a particular aspect of the invention of this disclosure, 225 Ac-labeled anti-CD33 antibody may be administered in one or more doses according to any of the dosing schedules enumerated herein until the subject receives a cumulative radiation dose of 1 μCi / kg of patient body weight, 2 μCi / kg of patient body weight, 3 μCi / kg of patient body weight, 4 μCi / kg of patient body weight, 5 μCi / kg of patient body weight, 6 μCi / kg of patient body weight, 7 μCi / kg of patient body weight, 8 μCi / kg of patient body weight, 9 μCi / kg of patient body weight, or 10 μCi / kg of patient body weight. According to certain aspects of the invention of this disclosure, an effective amount of anti-CD33 antibody may contain a radiation dose of 0.5 to 4 μCi / kg of patient body weight.

[0077] Generally, an effective dose of anti-CD33 antibody contains less than 16 mg / kg body weight, for example, 0.01 mg / kg to 10 mg / kg, or 0.01 mg / kg to 1.0 mg / kg, or 0.01 mg / kg to 0.1 mg / kg of protein.

[0078] According to a particular aspect of the present invention, the effective dose of the anti-CD33 antibody is the maximum tolerated dose (MTD) of the anti-CD33 antibody.

[0079] According to certain aspects of the invention disclosed herein, a radiolabeled monoclonal antibody can exhibit immunoreactivity to essentially the same antigen as a control monoclonal antibody, and the control monoclonal antibody includes an unlabeled monoclonal antibody to the same antigen epitope as the radiolabeled monoclonal antibody.

[0080] In exemplary embodiments of the invention disclosed herein, the anti-CD33 antibody is 225 The lintuzumab may be conjugated with a radiolabel such as Ac. The formulations of the present invention offer improved stability of radiolabeled antibodies, i.e., maintaining antibody stability for at least 24 hours (see WO2017 / 155937), but ultimately the radioisotope degrades the antibody. Therefore, an additional advantage of conjugating an anti-CD33 targeting agent to a radionuclide or radiotherapy agent disclosed herein is that such an agent can attenuate the antibody after it has reached target cells (cells expressing the CD33 antigen) but before it can damage normal cells.

[0081] The methods of the invention of the present disclosure further include the administration of a combination of two or more individual chemotherapeutic agents and immunotherapeutic agents, and the administration of a combination of two or more individual chemotherapeutic agents and immunotherapeutic agents. Therefore, the invention of the present disclosure envisions a method for treating proliferative disorders or disorders, comprising the administration of an antibody against CD33 and the administration of two or more individual chemotherapeutic agents, such as cladribine, cytarabine, mitoxantrone, and two or more of granulocyte colony-stimulating factor or filgrastim. In particular embodiments, the methods of the invention of the present disclosure include the administration of an antibody against CD33 and the administration of three or more individual chemotherapeutic agents, such as three or more of cladribine, cytarabine, mitoxantrone, and granulocyte colony-stimulating factor or filgrastim. The chemotherapeutic agents may be delivered together and / or separately during the course of treatment, according to a daily or hourly schedule specific to each chemotherapeutic agent.

[0082] The inventions of this disclosure further envision a method for treating proliferative disorders or disorders comprising the administration of an antibody against CD33 and the administration of a chemotherapy regimen such as CLAG-M. Such a combination of an immunotherapy agent and a chemotherapy regimen can provide therapeutic effects comparable to those of monotherapy, while mitigating the adverse side effects of monotherapy, and / or have improved efficacy that can be measured by a reduction in the total number of tumor cells, an increase in the time to recurrence, and other signs of the patient's health: for example, it can provide a synergistic effect superior to monotherapy (i.e., either the immunotherapy agent or the chemotherapy regimen).

[0083] The CLAG-M chemotherapy regimen includes the administration of a combination of chemotherapy agents: cladribine (Leustatin®), filgrastim (Neupogen®), cytarabine (Cytosar-U®), and mitoxantrone (Novantrone®).

[0084] Cladribine is an anti-cancer drug, an antimetabolite and a purine antagonist. Cladribine is structurally related to fludarabine and pentostatin, but has a different mechanism of action. Although the exact mechanism of action is not fully understood, evidence suggests that cladribine is phosphorylated by deoxycytidine kinase to the nucleotide cladribine triphosphate (CdATP; 2-chloro-2'-deoxyadenosine 5'-triphosphate), which aggregates and is incorporated into the DNA of cells such as lymphocytes containing high levels of deoxycytidine kinase and low levels of oxynucleotidase, resulting in DNA strand breakage and inhibition of DNA synthesis and repair. High levels of CDatP are also thought to inhibit ribonucleotide reductase, which leads to depletion of the triphosphate (DnTP) pool and subsequent inhibition of DNA strand breakage, DNA synthesis and repair, depletion of nicotinamide adenine dinucleotide (NAD) and ATP, and cell death. Unlike other antimetabolites, cladribine has a cytotoxic effect at rest and also stimulates lymphocyte proliferation. However, it is suggested that the cytotoxicity is associated with events important for cell entry into the S phase, as it causes cells to accumulate at the G1 / S junction. It also binds to purine nucleoside phosphorylase (PNP), but the relationship between this binding and the mechanism of action has not been established. Cladribine (2-CDa) can be administered at 3 - 7 mg / 1 m 2 of body surface area per subject per day, for example, 5 mg / 1 m 2 of body surface area per subject per day.

[0085] Cytarabine is an antimetabolite anti-cancer drug. Cytarabine acts through direct DNA damage and is incorporated into DNA. Cytarabine is cytotoxic to a wide variety of proliferating mammalian cells in culture. It exhibits cell phase specificity, mainly killing cells in DNA synthesis (S phase) and preventing cell progression from G1 to S phase under certain conditions. The mechanism of action is not fully understood, but it is thought that cytarabine acts through inhibition of DNA polymerase. Limited but significant incorporation of cytarabine into both DNA and RNA has also been reported. Cytarabine (Ara-C) is administered at 1 - 3 g / 1 m 2Body surface area, for example, 2g / 1m² of the target 2 It can be administered based on the body surface area per day.

[0086] Mitoxantrone is an anthracendione-derived anti-cancer drug. A DNA-reactive substance that intercalates into deoxyribonucleic acid (DNA) via hydrogen bonding, mitoxantrone causes crosslinking and strand disruption. Mitoxantrone also interferes with ribonucleic acid (RNA) and is a potent inhibitor of topoisomerase II, an enzyme involved in unwinding and repairing damaged DNA. Its cytotoxic effect on both proliferating and non-proliferating cultured human cells suggests a lack of cell cycle phase specificity. Mitoxantrone is administered at a dose of 5-15 mg / 1m². 2 Body surface area, for example, 10 mg / 1 m² of the target 2 It can be administered based on the body surface area per day.

[0087] Filgrastim is a recombinant, non-glycosylated form of granulocyte colony-stimulating factor (G-CSF), a 175-amino acid protein that induces neutrophil proliferation and maturation. Filgrastim binds to the G-CSF receptor and stimulates neutrophil production in the bone marrow. As a G-CSF analog, it controls the proliferation of involved progenitor cells and influences maturation into mature neutrophils. Filgrastim also stimulates the release of neutrophils from the bone marrow storage pool and shortens their maturation time. Filgrastim plays a role in increasing the phagocytic activity of mature neutrophils. In patients undergoing cytotoxic chemotherapy, filgrastim may accelerate neutrophil recovery and shorten the duration of the neutrophil phase. Filgrastim may be administered at doses of 100-500 ug / day, for example, 300 ug / day.

[0088] According to certain aspects of the invention disclosed herein, cladribine may be administered intravenously, cytarabine may be administered intravenously, mitoxantrone may be administered intravenously, and filgrastim may be administered subcutaneously.

[0089] According to certain embodiments of the methods of the invention described herein, the anti-CD33 antibody and the chemotherapy regimen may be administered simultaneously. Therefore, they may be provided in a single composition, or as several different compositions combined before administration, or administered within similarly short periods (e.g., within minutes or hours of each other). Alternatively, the anti-CD33 antibody and the chemotherapy regimen may be administered sequentially. Therefore, the anti-CD33 antibody may be administered before the chemotherapy regimen, after the chemotherapy regimen, or both before and after the chemotherapy regimen. Furthermore, the chemotherapy regimen may be administered before the anti-CD33 antibody, before the anti-CD33 antibody, or both before and after the anti-CD33 antibody.

[0090] According to a particular aspect of the method of the present invention, the anti-CD33 antibody may be administered throughout the entire treatment period according to a dosing schedule selected from the group consisting of once every 5, 7, 10, 12, 14, 20, 21, 24, 28, 30, 35, and 42 days, the treatment period comprising at least two doses.

[0091] According to certain aspects of the invention disclosed herein, the anti-CD33 targeting agent may be administered in two doses, for example, on day 1 and day 5, 6, 7, 8, 9, or 10 of the treatment period, or on day 1 and day 8 of the treatment period, according to a dosing schedule.

[0092] According to certain aspects of the invention disclosed herein, the chemotherapy treatment regimen may be administered according to a dosing schedule selected from once every 1, 2, 3, 4, 5, or 6 days throughout the entire treatment period, and the treatment period includes at least two dosings.

[0093] According to certain aspects of the invention of this disclosure, the standard administration protocol for the CLAG-M chemotherapy regimen includes intravenous administration of cladribine over a period of approximately 2 hours, followed by intravenous administration of cytarabine over a period of approximately 4 hours. The administration of cytarabine may be approximately 2 hours after the administration of cladribine. These two drugs may be administered daily for 5 days. Furthermore, mitoxantrone may be administered intravenously on days 1 to 3 of the administration protocol. Finally, filgrastim may be administered subcutaneously on days 0 to 5 of the administration protocol.

[0094] According to certain aspects of the invention of this disclosure, the immunotherapy agent (anti-CD33 antibody) may be administered on any one day from day 5 to day 30 of the standard dosing protocol of the CLAG-M chemotherapy regimen. For example, the anti-CD33 antibody may be administered on days 9 to 20 of the standard dosing protocol of the CLAG-M chemotherapy regimen, or on days 12, 13, 14, 15, or 16 of the standard dosing protocol of the CLAG-M chemotherapy regimen, or on day 14 of the standard dosing protocol of the CLAG-M chemotherapy regimen.

[0095] According to certain aspects of the invention of this disclosure, the method may include the administration of a labeled monoclonal antibody against CD33 in one fraction and an unlabeled (e.g., "naked") fraction. The unlabeled fraction may contain the same antibody against the same epitope as the labeled fraction. In this way, the overall antibody concentration can be altered, while the total radioactivity of the antibody can be reduced or set. For example, the total protein concentration of the administered unlabeled antibody fraction may vary depending on the exact nature of the disease being treated, the patient's age and weight, the identity of the monoclonal antibody, and the label (e.g., radionuclide) selected for labeling the monoclonal antibody.

[0096] According to certain aspects of the invention disclosed herein, multiple doses of an immunotherapy agent (anti-CD33 antibody) may be administered. Furthermore, each dose may be the same or different. For example, the first dose of the immunotherapy agent (induction dose) may be greater than additional doses of the immunotherapy agent (sequential administration).

[0097] In certain embodiments, the methods of the inventions of this disclosure include the administration of monospecific and / or multispecific immunological agents and the chemotherapy regimen CLAG-M, and may further include the administration of one or more additional therapeutic agents. The additional therapeutic agents may be related to the disease or condition being treated. Such administrations may be concurrent, separate, or sequential with the administration of effective amounts of immunotherapeutic agents and the CLAG-M chemotherapy regimen as detailed herein. In the case of concurrent administration, the agents may be administered as a single composition or as separate compositions, as necessary.

[0098] Exemplary additional therapeutic agents include, at a minimum, chemotherapeutic agents, anti-inflammatory agents, immunosuppressants, immunomodulators, or combinations thereof. Furthermore, one or more additional therapeutic agents may include antimyeloma agents such as dexamethasone, melphalan, doxorubicin, bortezomib, lenalidomide, prednisone, carmustine, etoposide, cisplatin, vincristine, cyclophosphamide, and thalidomide.

[0099] Examples of chemotherapeutic agents include antimetabolites such as methotrexate, 6-mercaptopurine, 6-thioguanine, fludarabine, 5-fluorouracil, dacarbazine, hydroxyurea, asparaginase, gemcitabine, and similar drugs.

[0100] Examples of chemotherapeutic agents include alkylating agents such as mechloretamine, thiotepa, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, dacarbazine (DTIC), procarbazine, mitomycin C, cisplatin and carboplatin, and other platinum derivatives, as well as similar drugs.

[0101] Examples of chemotherapeutic agents include antibiotics such as dactinomycin (formerly actinomycin), bleomycin, calitiamycin, daunorubicin (formerly daunomycin), doxorubicin, idarubicin, mitramycin, mitomycin, mitoxantrone, plicamycin, anthramycin (AMC), and similar drugs.

[0102] Similar chemotherapeutic agents include, for example, taxanes such as docetaxin and paclitaxel, and mitotic inhibitors such as vinca alkaloids, such as vindesine, vincristine, vinblastine, and vinorelbine.

[0103] Examples of chemotherapeutic agents include topoisomerase inhibitors such as topotecan.

[0104] Exemplary chemotherapeutic agents include, for example, growth factor inhibitors such as ErBb1 (EGFR) inhibitors (gefitinib (Iressa®), cetuximab (Erbitux®), erlotinib (Tarceva®), HuMax-EGFr (2F8 disclosed in WO2002 / 100348) and similar agents), and ErBb2 (Her2 / neu) inhibitors (transtuzumab (Herceptin®) and similar agents). In one embodiment, such growth factor inhibitors may be farnesyltransferase inhibitors such as SCH-66336 and R115777. In one embodiment, such growth factor inhibitors may be vascular endothelial growth factor (VEGF) inhibitors such as bevacizumab (Avastin®).

[0105] Exemplary chemotherapeutic agents include tyrosine kinase inhibitors such as imatinib (Glivec, Gleevec ST1571), lapatinib, PTK787 / ZK222584, and similar drugs.

[0106] Exemplary chemotherapeutic agents include histone deacetylase inhibitors. Examples of such histone deacetylase inhibitors include hydroxamic acid hybrid compounds such as SAHA (subloanilide hydroxamic acid).

[0107] Exemplary chemotherapeutic agents include P38a MAP kinase inhibitors such as SCIO-469.

[0108] Exemplary chemotherapeutic agents include inhibitors of angiogenesis, neoangiogenesis, and / or other forms of angiogenesis. Examples of such inhibitors include urokinase inhibitors, matrix metalloproteinase inhibitors (such as Marimast, Neovastat, BAY 12-9566, AG 3340, BMS-275291, and similar agents), and inhibitors of endothelial cell migration and proliferation (such as TNP-470, squalamine, 2-methoxyestradiol, combretastatin, endostatin, angiostatin, penicylamine, SCH66336 (Schering-Plough Corp, Madison, NJ), R115777 (Janssen)). Pharmaceutica, Inc. (Titusville, NJ) and similar drugs, etc., angiogenic growth factor antagonists (ZD6474, SU6668), antibodies against angiogenic substances and / or their receptors (VEGF, bFGF, and angiopoietin-1, etc.), thalidomide (Thalomid®), thalidomide derivatives (CC-5013 (Lenalidomide, Revlimid®, etc.) and CC4047 (Actimid®), Sugen Examples include 5416, SU5402, anti-angiogenic ribozymes (angiozymes, etc.), interferon α (interferon α2α, etc.), suramin and similar drugs), VEGF-R kinase inhibitors and other anti-angiogenic tyrosine kinase inhibitors (SU011248, etc.), endothelium-specific integrin / survival signaling inhibitors (vitaxin and similar drugs, etc.), copper antagonists / chelators (tetrathiomolybdate, captopril and similar drugs, etc.), carboxamide triazole (CAI), ABT-627, CM101, interleukin-12 (IL-12), IM862, PNU145156E, and nucleotide molecules that inhibit angiogenesis (antisense-VEGF-cDNA, cDNA encoding angiostatin, cDNA encoding p53, and cDNA encoding deficient VEGF receptor-2) and similar drugs.

[0109] Other examples of such inhibitors of angiogenesis, neoangiogenesis, and / or other forms of angiogenesis include anti-angiogenic heparin derivatives and related molecules (e.g., heparinase III), tetozolomide, NK4, macrophage migration inhibitors (MIFs), cyclooxygenase-2 inhibitors, hypoxia-inducible factor 1 inhibitors, anti-angiogenic soy isoflavones, ortipraz, fumagiline and its derivatives, somatostatin derivatives, pentosan polysulfate, tecogalan sodium, dalteparin, tumstatin, thrombospondin, NM-3, combrestatin, canstatin, avastatin, antibodies against other related targets (anti-α-v / β-3 integrin anti-quininostatin mAbs, etc.) and similar agents.

[0110] Exemplary chemotherapeutic agents include thalidomide (Thalomid®), thalidomide analogs (CC-5013 (Lenalidomide, Revlimid®) and / or CC4047 (Actimid®)).

[0111] Exemplary chemotherapeutic agents may include additional antibody therapies or proteasome inhibitors, such as bortezomib (Velcade®), corticosteroids, such as prednisone, prednisolone, dexamethasone, etc., and bisphosphonates. Potentially suitable examples of bisphosphonates include pamidronate (Aredia®), zoledronic acid (Zometa®), clodronate (Bonefos®), lysendronate (Actonel®), ibandronate (Boniva®), etidronate (Didronel®), alendronate (Fosamax®), tildronate (Skelid®), incadronate (Yamanouchi Pharmaceutical), and minodronate (YM529, Yamanouchi).

[0112] Exemplary chemotherapeutic agents include erythropoiesis agents. Examples of suitable erythropoiesis agents include erythropoietins (EPOs) such as epoetin alfa (e.g., Procrit®, Epogen®, and Eprex®) and epoetin beta (e.g., NeoRecormon®), as well as erythropoiesis-promoting proteins (e.g., Aranesp®).

[0113] Exemplary chemotherapeutic agents include anti-anergistic agents (e.g., small molecule compounds, proteins, glycoproteins, or antibodies that disrupt tolerance to tumor and cancer antigens).

[0114] Exemplary chemotherapeutic agents include viruses, viral proteins, etc. Replication-deficient viruses that are typically capable of one or only a few replications in vivo and that target tumor cells may be useful components of such compositions and methods, for example. Such viral agents may include, or be associated with, nucleic acids encoding immunostimulants such as GM-CSF and / or IL-2. Both naturally occurring oncolytic viruses and such recombinant oncolytic viruses (e.g., HSV-1 virus, reovirus, replication-deficient and replication-susceptible adenoviruses) may be useful components of such methods and compositions.

[0115] Exemplary anti-inflammatory agents may be selected from steroidal drugs and NSAIDs (non-steroidal anti-inflammatory drugs). Other anti-inflammatory agents include aspirin and other salicylates, Cox-2 inhibitors (e.g., rofecoxib and celecoxib), NSAIDs (e.g., ibuprofen, fenoprofen, naproxen, sulindac, diclofenac, piroxicam, ketoprofen, diflunisal, nabumeton, etodrug, oxaprozin, and indomethacin), anti-IL6R antibodies, anti-IL8 antibodies, anti-IL15 antibodies, anti-IL15R antibodies, anti-CD4 antibodies, anti-CD11a antibodies (e.g., Efa The following may be selected from lizumab, anti-α4 / β-1 integrin (VLA4) antibodies (e.g., natalizumab), CTLA4-1g for the treatment of inflammatory diseases, disease-modifying antirheumatic drugs (DMARDs) such as prednisolone, prednisone, and methotrexate, sulfasalazine, pyrimidine synthesis inhibitors (e.g., leflunomide), IL-1 receptor blockers (e.g., anakinra), TNF-α blockers (e.g., etanercept, infliximab, and adalimumab), and similar drugs.

[0116] Examples of immunosuppressants and / or immunomodulators include corticosteroids such as cyclosporine, azathioprine, mycophenolic acid, mycophenolate mofetil, and prednisone, methotrexate, gold salts, sulfasalazine, antimalarial drugs, Brekinal, leflunomide, mizoribine, 15-deoxysperguarine, 6-mercaptopurine, cyclophosphamide, rapamycin, tacrolimus (FK-506), OKT3, antithymocyte globulin, thymopentin, thymosin-α, and similar drugs.

[0117] According to certain aspects of the inventions of this disclosure, additional therapeutic agents may include antimyeloma agents. Exemplary antimyeloma agents include dexamethasone, melphalan, doxorubicin, bortezomib, lenalidomide, prednisone, carmustine, etoposide, cisplatin, vincristine, cyclophosphamide, and thalidomide, some of which are shown above as chemotherapeutic agents, anti-inflammatory agents, or immunosuppressants.

[0118] According to certain aspects of the invention of the present disclosure, the additional therapeutic agent may include allopurinol, which is orally administered at a dose of 300 to 600 mg / day starting on the first day of the treatment period and continued until at least 7 days after the CD33 targeting agent. For patients with an absolute neutrophil count of less than 500 / ul, prophylactic antibiotics and antifungal therapies can be included. To reduce infusion-related reactions, analgesics and antihistamines may also be included before the administration of the CD33 targeting agent by infusion.

[0119] The additional therapeutic agent can be administered according to any standard regimen known in the art. For example, the therapeutic agent may be administered at a concentration in the range of 1 to 500 mg / m 2 , and the amount is calculated as a function of the patient's surface area (m 2 ). For example, an exemplary dose of paclitaxel may include 15 mg / m 2 to 275 mg / m 2 , an exemplary dose of docetaxel may include 60 mg / m 2 to 100 mg / m 2 , an exemplary dose of epothilone may include 10 mg / m 2 to 20 mg / m 2 , and an exemplary dose of calicheamicin may include 1 mg / m 2 to 10 mg / m 2 . Exemplary doses are listed herein, but they are provided for reference only and are not intended to limit the dosage range of the agents of the invention of the present disclosure.

[0120] The method of the invention of the present disclosure further includes transplanting autologous or allogeneic stem cells to the subject after the administration of the anti-CD33 antibody. When the anti-CD33 antibody is labeled with a radionuclide, the stem cells can be transplanted 8 to 20 days after the administration of the anti-CD33 antibody, or further 10 to 16 days after the administration of the anti-CD33 antibody, when the radiation dose from the anti-CD33 antibody is not harmful to the transplanted cells.

[0121] The inventions of this disclosure provide methods for treating subjects having proliferative disorders, methods for inhibiting the growth and / or proliferation of CD33-expressing cells, and methods for treating diseases or disorders involving CD33-expressing cells. Proliferative disorders can be hematological malignancies.

[0122] Accordingly, the invention of this disclosure provides a method for treating subjects having proliferative disorders. In particular cases, the proliferative disorder may be a hematological malignancy. According to a particular aspect of the invention of this disclosure, the hematological malignancy may be one or more of multiple myeloma, acute myeloid leukemia, myelodysplastic syndrome, and myeloproliferative neoplasm. According to a particular aspect of the invention, the CD33-expressing cells are multiple myeloma cells.

[0123] In particular aspects of the invention of this disclosure, a method is provided for treating a target solid tumor, the method comprising administering a therapeutically effective amount of a monoclonal antibody detailed herein (i.e., an anti-CD33 antibody, and / or a bispecific antibody against the epitopes of CD33 and another antigen detailed above) to the target in combination with the chemotherapy regimen CLAG-M. While not intending to be constrained by any particular theory, based on the immunomodulatory effects observed with the anti-CD33 monoclonal antibody described herein, the method of the invention of this disclosure may be effective in treating solid tumors. The treatment may include intravenous, intraperitoneal, or intratumoral infusion of the immunotherapy agent.

[0124] The following aspects are disclosed in this application.

[0125] Embodiment 1. A method for inhibiting the growth and / or proliferation of CD33-expressing cells, comprising administering an effective amount of an anti-CD33 targeting agent and an effective amount of a combination of two or more individual chemotherapeutic agents to the target.

[0126] Embodiment 2. The method according to Embodiment 1, wherein the combination of two or more individual chemotherapeutic agents comprises two or more of cladribine, cytarabine, mitoxantrone, and filgrastim.

[0127] Embodiment 3. The method according to either Embodiment 1 or 2, wherein the combination of two or more individual chemotherapeutic agents comprises two or more of the following: cladribine administered intravenously, cytarabine administered intravenously, mitoxantrone administered intravenously, and filgrastim administered subcutaneously.

[0128] Apparatus 4. The effective dose of a combination of two or more individual chemotherapeutic agents is 3-7 mg of cladribine / 1 mg of the target. 2 Body surface area / day, 1-3g of cytarabine / 1 m² of the target area 2 Body surface area / day, 5-15 mg of mitoxantrone / target 1 m² 2 The method according to any one of embodiments 1 to 3, comprising two or more of the following: body surface area / day and 100-500 ug of filgrastim / day.

[0129] Applicable aspect 5. The effective dose of a combination of two or more individual chemotherapeutic agents is 5 mg cladribine / 1 mg of the target. 2 Body surface area / day, 2g of cytarabine / 1 m² of the target area 2 Body surface area / day, 10 mg of mitoxantrone / target area 1 m 2 The method according to any one of embodiments 1 to 4, comprising body surface area / day and 300 ug of filgrastim / day.

[0130] Embodiment 6. The method according to any one of Embodiments 2 to 5, wherein the effective dose of the combination is administered to the subject as follows: cladribine on days 1, 2, 3, 4, and 5 of the treatment period; cytarabine on days 1, 2, 3, 4, and 5 of the treatment period; mitoxantrone on days 1, 2, and 3 of the treatment period; and filgrastim on days 0, 1, 2, 3, 4, and 5 of the treatment period.

[0131] Embodiment 7. The method according to any one of Embodiments 1 to 6, wherein the anti-CD33 targeting agent is administered on any one day from day 1 to day 30 of the treatment period, or on any one day from day 9 to day 20 of the treatment period, or on day 12, day 13, day 14, day 15, or day 16 of the treatment period, or on day 14 of the treatment period.

[0132] Embodiment 8. The method according to any one of Embodiments 1 to 7, wherein an anti-CD33 targeting agent and a chemotherapeutic agent are administered sequentially.

[0133] Embodiment 9. The method according to any one of Embodiments 1 to 8, wherein the anti-CD33 targeting agent is administered before the chemotherapeutic agent, or the anti-CD33 targeting agent is administered after the chemotherapeutic agent.

[0134] Embodiment 10. The anti-CD33 targeting agent comprises a radiolabeled antibody against CD33, wherein the radiolabeling is 131 I, 125 I, 123 I, 90 Y, 177 Lu, 186 Re, 188 Re, 89 Sr, 153 Sm, 32 P, 225 Ac, 213 Bi, 213 Po, 211 At, 212 Bi, 213 Bi, 223 Ra, 227 Th, 149 Tb, 137 Cs, 212 Pb or 103 The method according to any one of embodiments 1 to 9, selected from Pd, or a combination thereof.

[0135] Embodiment 11. The method according to any one of Embodiments 1 to 10, wherein the anti-CD33 targeting agent comprises lintuzumab, gemtuzumab, vadasutuximab, or a combination thereof.

[0136] Embodiment 12. The method according to any one of Embodiments 1 to 11, wherein the anti-CD33 targeting agent comprises HuM195.

[0137] Embodiment 13. Anti-CD33 targeting agents are 225The method according to any one of embodiments 1 to 12, wherein the anti-CD33 targeting agent is labeled with Ac and the effective amount of the anti-CD33 targeting agent includes a radiation dose of 0.1 to 10 uCi / kg of body weight of the target, or 0.2 to 8 uCi / kg of body weight of the target, or 0.5 to 4 uCi / kg of body weight of the target, or 0.2 to 2.0 uCi / kg of body weight of the target.

[0138] Apparatus 14. Anti-CD33 targeting agent 131 The method according to any one of embodiments 1 to 13, wherein the effective amount of the anti-CD33 targeting agent is labeled with I and includes a radiation dose of 10 mCi to 200 mCi, or 50 mCi to 150 mCi, or 50 mCi to 100 mCi.

[0139] Embodiment 15. The method according to any one of Embodiments 1 to 14, wherein the effective dose of the anti-CD33 targeting agent comprises a protein dose of less than 16 mg / kg of body weight of the target, less than 10 mg / kg of body weight of the target, or less than 6 mg / kg of body weight of the target, less than 500 μg / kg of body weight of the target, less than 100 μg / kg of body weight of the target, less than 50 μg / kg of body weight of the target, or less than 20 μg / kg of body weight of the target.

[0140] Apparatus 16. The method according to any one of Apparatus 1 to 15, which can treat one or more hematological diseases or disorders selected from the following: acute lymphoblastic leukemia, acute myeloid leukemia, chronic myelomonocytic leukemia, erythrocytic leukemia, acute megakaryoblastic leukemia, histiocytic leukemia, myelosarcoma, mast cell proliferation disorder, and myelodysplastic syndrome.

[0141] Embodiment 17. The method according to any one of Embodiments 1 to 16, wherein the method can treat a relapsed / refractory hematological disorder or disability, the hematological disorder or disability being selected from multiple myeloma, acute myeloid leukemia, myelodysplastic syndrome, and myeloproliferative neoplasm.

[0142] Embodiment 18. The method according to any one of Embodiments 1 to 17, wherein the cells expressing CD33 are blast cells such as myeloblast cells or malignant plasma cells. [Examples]

[0143] Example 1: CD33 specificity and stability Actinium-225 (Ac 225 Lintuzumab conjugated with ) was tested for cytotoxicity against specific cell types expressing CD33. For example, a suspension of HL60 (leukocytes) was tested with various doses of radiolabeled lintuzumab (lintuzumab-Ac). 225 When incubated with the dose (LD50), 50% of the cells were killed. 50 It was found that the concentration was 8 Pci per 1 mL of cell suspension.

[0144] In a study to explore the reactivity of radiolabeled lintuzumab with peripheral blood and bone marrow cells from non-human primates and human frozen tissues, radiolabeled lintuzumab showed reactivity only to mononuclear cells, demonstrating its specificity. Furthermore, in a study to determine the stability of the radiolabeling on the antibody, 300 nCi of radiolabeled lintuzumab (0.12 ml) was injected into the tail of 10 normal mice (8-week-old female Balb / c mice from Taconic, Germantown, and New York). Serum samples collected over 5 days showed that actinium-225 remained bound to lintuzumab, demonstrating the in vivo stability of the radiolabeling on the antibody.

[0145] The maximum tolerated dose (MTD) for a single injection of radiolabeled lintuzumab was determined to be 3 Uci / patient body weight kg. For divided doses (e.g., two equal doses administered over 4–7 days), the MTD was determined to be 2 Uci / kg per dose, or a total of 4 Uci / kg. This data was determined by injections in patients with relapsed / refractory AML: 21 patients received escalating doses of radiolabeled lintuzumab (0.5 Uci / kg–4 Uci / kg). MTD determination was based on the severity of adverse effects observed at each dose level. Antileukemic effects included peripheral blood blast removal in 13 of 19 evaluable patients. Myeloblast reduction was observed in 12 of 18 evaluable patients at 4 weeks post-treatment (including 9 showing a reduction of ≥50%). Three patients treated with 1 uCi / kg, 3 uCi / kg, and 4 uCi / kg, respectively, had ≤5% blast cells after treatment.

[0146] Example 2: Maximum human tolerance and efficacy of CD33 In the Phase I trial, lintuzumab-Ac 225 This is used to determine the MTD of divided doses and subsequent supportive therapy with granulocyte colony-stimulating factor (GCSF). A typical cycle is approximately 42 days. The cycle consists of lintuzumab-Acid on day 1. 225 Treatment is initiated with divided doses of lintuzumab Acetate (GCSF) on day 9, and GCSF is continued according to appropriate medication instructions until the absolute neutrophil count (ANC) exceeds 1,000 (expected to occur within 5-10 days). Peripheral blood is assessed for paraprotein loading on days 14, 21, 28, 35, and 42. Bone marrow aspiration is performed on day 42 to assess plasma cell infiltration. If the response on day 42 is partial or better than before but not complete, and the patient is otherwise eligible, the patient is readministered in a new cycle at the same dose level from day 1 to day 60 of the first cycle. If no dose-limiting toxicity is observed, the patient is given 4 μCi / kg of lintuzumab Acetate (Acetate Acetate). 225 The cycle continues to use the above algorithm until it receives the cumulative dose.

[0147] Example 3: Maximum CD33 tolerance when CLAG-M is added The CLAG-M treatment regimen consists of the following medications: cladribine 5 mg / m² administered over 2 hours from day 1 to day 5. 2 IV; Cytarabine 2 g m / m² administered over 4 hours from day 1 to day 5. 2 IV (start 2 hours after cladribine infusion is complete); mitoxantrone 10 mg / m² on days 1-3. 2 IV; G-CSF (filgrastim) at a dose of 300 μg from day 0 to day 5. Lintuzumab-Ac 225 It is administered as an IV infusion over 30 minutes on day 14 of treatment. Lintuzumab-Ac for 3 trial cohorts 225 The dosage of 225 is described below. Prophylaxis for radioactive nephrotoxicity is performed using oral spironolactone for up to one year after treatment. [Table 1]

[0148] Monitoring and response assessment: At baseline (if relapse is diagnosed) and after CLAG-M therapy, lintuzumab-Ac 225A bone marrow biopsy should be performed within 42 days after administration of the drug, or when the blood cell count recovers, whichever comes first. The patient's complete blood count and metabolic panel should be monitored daily during hospitalization. The response to treatment should be evaluated according to the standard definitions developed by the International Working Group, with complete remission defined as myeloblasts <5%, absolute neutrophil count ≥1000 / L, and platelet count ≥100,000 / L. Complete remission (CR) without platelet recovery is defined as CRp, and complete remission (CR) without white blood cell recovery is defined as CRi. Partial remission (PR) is defined as a reduction in the percentage of blasts in the bone marrow from at least 50% to 5-25% and normalized blood cell count. Patients who do not achieve CR, CRi, CRp, or PR are defined as ineffective (NR). Progression-free survival (PFS) is calculated from day 1 of remission until relapse is confirmed. Overall survival (OS) is calculated from day 1 of salvage therapy until death. Hematological and excessive hematological toxicity will be assessed according to CTCAE criteria. DLTs include subsequent events unless they are caused by non-treatment alternatives.

[0149] Example 4: CD33 combination therapy using chemotherapeutic agents In the Phase I trial, 18 patients with relapsed AML were administered in divided doses on days 1 and 8 in combination with low-dose Ara-C (LDAC). 225 Treatment with Ac-lintuzumab was used. The treatment was found to induce remission in elderly patients with untreated AML at doses exceeding 0.5 uCi / kg / fraction. In the phase 2 portion of the clinical trial, 13 patients with early symptoms of AML who were deemed unsuitable for cytotoxic chemotherapy were given divided doses on days 1 and 8 without LDAC. 225 Treatment with Ac-lintuzumab was performed. Preliminary data are available for median age 75 years (range: 65–82) and median PS 2 (2 patients 0–1, 3 patients 2, and 2 patients 3). Six patients (67%) had previously received treatment for AHD (5 with MDS, 1 with atypical CML). At baseline, 5 patients had ANC ≥ 500 / μL, only 2 had ANC ≥ 1000 / μL, and only 1 had platelet > 50,000 / μL.

[0150] In all three patients, myelosuppression, including grade 4 thrombocytopenia with myelopyelopathy, was observed for more than 6 weeks post-treatment. The only non-hematological toxicities reported as grade 3 or higher with a score of 1 or higher were pneumonia and cellulitis. No venous occlusion occurred. The 30-day mortality rate was 33% (disease progression, acute or chronic respiratory failure, and traumatic intracranial hemorrhage after falls).

[0151] The target response was observed in 5 out of 9 patients (56%): 2 patients achieved complete remission (CRp) with incomplete platelet count recovery, and 3 patients achieved complete remission (CRi) with incomplete hematological recovery. Two patients had resistant disease.

[0152] The median neutrophil recovery (ANC ≥ 500 / μl) is: 225 The timeframe was 36 days (range: 20-60) from the first dose of Ac-lintuzumab. Two patients with CRp recovered neutrophil counts on days 60 and 36. Two patients with CRi did not reach ANC ≥ 500 / μL when the infectious period ended on days 65 and 56, and the third patient's infectious period ended beyond day 66 without ANC recovery. Patients without pre-existing hematological disorders (AHD) may not be able to recover to normal neutrophil production, so patients without AHD may be more beneficial. Of the three patients without AHD, one recovered ANC on day 36, one died from the infection on day 56 without ANC recovery, and one had not shown ANC recovery results beyond day 66. No patients reached platelet count > 20,000 / μL without transfusion.

[0153] 2 μCi / kg / fraction 225 Preliminary data from this phase 2 trial of Ac-lintuzumab monotherapy confirmed a 56% remission rate in elderly patients, many of whom had AHD, who were unsuitable for more aggressive treatments. Myelosuppression at this dose was considered to be longer than acceptable in this population, but additional trials will be continued at 1.5 μCi / kg / fraction to shorten the recovery time.

[0154] Example 5: CD33 combination therapy using CLAG-M In the Phase I clinical trial, patients will be hospitalized for administration of the CLAG-M chemotherapy agent. Baseline characteristics will be obtained, including age, sex, race, AML subtype, date of initial diagnosis, date of relapse, interval between diagnosis and relapse, cytogenetic and molecular mutations at diagnosis, previous chemotherapy or human stem cell transplantation cycles (HSCT), baseline clinical examinations at admission, and percentages of blasts in peripheral blood and bone marrow at diagnosis and relapse. The CLAG-M chemotherapy regimen will be administered as described herein. During hospitalization, the patient's complete blood count and metabolic panel will be monitored daily. The bone marrow (BM) test obtained at the time of diagnosis of RR-AML will be used as baseline. When the absolute neutrophil count (ANC) recovers to >1000 cells / cu.mm, or when lintuzumab-Ac 225 A repeated BM trial will be obtained after completion of treatment, either up to 42 days (or beyond 49 days) after administration, whichever comes first. The response to treatment will be evaluated according to the standard definition developed by the International Working Group. Patients who respond to treatment may be considered for allogeneic stem cell transplantation later. Patients who show a partial response (PR) or are ineffective may be given additional chemotherapeutic agents at the discretion of the treating physician and the patient's tolerance. Hematological and excessive hematological toxicity will be evaluated according to the Common Terminology Criteria for Adverse Events, version 4.0.

[0155] Dose escalation will be carried out according to a 3+3 design. CLAG-M chemotherapy drugs will be administered at fixed doses and schedules. Lintuzumab-Ac 225 The initial dose is 0.25 uCi / kg (dose level 1), and the highest dose administered is 0.75 uCi / kg.

[0156] The dose-limiting toxicity (DLT) observation period for dose escalation is one cycle. The first patient at each new dose level must be observed for one cycle for the occurrence of adverse events (AEs) before the second patient is treated at that dose level. Patients then proceed sequentially to each dose level.

[0157] At each dose level, if the first three patients at that level do not experience a DLT (Drug Limitation Tolerance), new patients can be advanced to the next higher dose level. If one of the three patients experiences a DLT, up to three patients will be treated at the same dose level. If an additional three patients at that dose level do not experience a DLT, new patients can be introduced to the next higher dose level. However, if one or more of the additional three patients experience a DLT, no further patients are initiated at that dose level, and the previous dose is considered the MTD (Mean Time Tolerance). The MTD is defined as the highest dose level at which none of the first three treated patients, or only one of the six first treated patients, experience a DLT. Preferred embodiments of the present invention are as follows: 1. A method for inhibiting the growth and / or proliferation of cells expressing CD33, A method characterized by administering an effective amount of an anti-CD33 targeting agent and an effective amount of a combination of two or more individual chemotherapeutic agents to a target. 2. The method according to 1 above, wherein the combination of two or more individual chemotherapeutic agents comprises two or more of the following: cladribine, cytarabine, mitoxantrone, and granulocyte colony-stimulating factor or filgrastim. 3. The effective amount of the combination of the two or more individual chemotherapeutic agents is 3-7 mg of cladribine / 1 m of the above subject 2 Body surface area / day, 1-3 g of cytarabine / 1 ml of the above-mentioned substance 2 Body surface area / day, 5-15 mg of mitoxantrone / 1 mg of the above-mentioned subject 2 Body surface area / day, and The method described in item 2 above, comprising two or more of the following: 100-500 ug of filgrastim / day. 4. The effective amount of the combination of the two or more individual chemotherapeutic agents is 5 mg of cladribine / 1 mg of the above subject 2 Body surface area / day, 2g cytarabine / 1m of the subject 2 Body surface area / day, 10 mg mitoxantrone / 1 m of the above subject 2 Body surface area / day, and The method according to item 2 above, comprising 300 ug of filgrastim / day. 5. The effective amount of the combination of the two or more individual chemotherapeutic agents is The method according to 3 or 4 above, wherein the subjects are administered cladribine on days 1, 2, 3, 4, and 5 of the treatment period, cytarabine on days 1, 2, 3, 4, and 5 of the treatment period, mitoxantrone on days 1, 2, and 3 of the treatment period, and filgrastim on days 0, 1, 2, 3, 4, and 5 of the treatment period. 6. The method according to 5, wherein the anti-CD33 targeting agent is administered on one of the following days: the 5th to 30th day of the treatment period, the 9th to 20th day of the treatment period, or the 14th day of the treatment period. 7. The method according to claim 1, wherein the combination of the anti-CD33 targeting agent and the two or more individual chemotherapeutic agents is administered sequentially. 8. The method according to 7, wherein the anti-CD33 targeting agent is administered before the combination of the two or more chemotherapeutic agents, or the anti-CD33 targeting agent is administered after the combination of the two or more chemotherapeutic agents. 9. The anti-CD33 targeting agent comprises a radiolabeled antibody against CD33, wherein the radiolabeling is 131I、 125 I、 123 I、 90 Y、 177 Lu, 186 Re, 188 Re, 89 Sr、 153 Sm, 32 P、 225 Ac, 213 Bi, 213 Po, 211 At, 212 Bi, 213 Bi, 223 Ra, 227 Th, 149 Tb, 137 Cs, 212 Pb or 103 The method described in 1 above, selected from Pd, or a combination thereof. 10. The method according to 9 above, wherein the anti-CD33 targeting agent comprises lintuzumab, gemtuzumab, vadasutuximab, or a combination thereof. 11. The method according to 1 above, wherein the anti-CD33 targeting agent comprises HuM195. 12. The anti-CD33 targeting agent is 225 The method according to 11, wherein the anti-CD33 targeting agent is labeled with AC, and the effective amount of the anti-CD33 targeting agent includes a radiation dose of 0.1 to 10 uCi / kg of body weight of the target, or 0.2 to 8 uCi / kg of body weight of the target, or 0.5 to 4 uCi / kg of body weight of the target, or 0.2 to 2.0 uCi / kg of body weight of the target. 13. The anti-CD33 targeting agent is 131 The method according to 11, wherein the anti-CD33 targeting agent is labeled with I, and the effective amount of the anti-CD33 targeting agent includes a radiation dose of 10 mCi to 1200 mCi, or 20 mCi to 200 mCi, or 50 mCi to 150 mCi. 14. The method according to 12 or 13, wherein the effective amount of the anti-CD33 targeting agent comprises a protein dose of less than 16 mg / kg of body weight of the subject, less than 10 mg / kg of body weight of the subject, less than 6 mg / kg of body weight of the subject, less than 50 μg / kg of body weight of the subject, or less than 20 μg / kg of body weight of the subject. 15. The method according to item 1 above, which can treat a hematological disorder or condition selected from one or more of the following: multiple myeloma, acute myeloid leukemia, myelodysplastic syndrome, and myeloproliferative neoplasm. 16. The method according to item 1 above, which can treat relapsed / refractory acute myeloid leukemia. 17. The method according to item 1 above, wherein the cells expressing CD33 are myeloblast cells or blast cells such as malignant plasma cells. 18. Methods for treating hematological diseases or disorders, The procedure involves administering to a subject with the aforementioned hematological disorder or disability an effective dose of a radiolabeled anti-CD33 targeting agent and an effective dose of a chemotherapy regimen containing cladribine, cytarabine, mitoxantrone, and filgrastim. The aforementioned radiolabeled anti-CD33 targeting agent 225 A method characterized by comprising compositions of AC-labeled HuM195 and unlabeled HuM195. 19. The method according to 18 above, wherein the hematological disorder or disability is relapsed / refractory acute myeloid leukemia. 20. 225 The composition comprising Ac-labeled HuM195 and unlabeled HuM195 is A radiation dose of 0.1~10 uCi / kg of the subject's body weight, or 0.2~8 uCi / kg of the subject's body weight, or 0.5~4 uCi / kg of the subject's body weight, or 0.2~2.0 uCi / kg of the subject's body weight, The method according to 18 above, comprising a protein dose of less than 16 mg / kg of body weight of the subject, less than 10 mg / kg of body weight of the subject, less than 6 mg / kg of body weight of the subject, less than 50 μg / kg of body weight of the subject, or less than 20 μg / kg of body weight of the subject.

Claims

1. A pharmaceutical composition for inhibiting the growth and / or proliferation of CD33-expressing cells in a human subject, which is combined with a combination of four individual chemotherapeutic agents, wherein the pharmaceutical composition comprises an anti-CD33 targeting agent. The four individual chemotherapeutic agents mentioned above are as follows: 3-7 mg of cladribine / 1 m of the above subject 2 Body surface area / day, 1-3 g of cytarabine / 1 m of the above-mentioned subject 2 Body surface area / day, 5-15 mg of mitoxantrone / 1 m of the above subject 2 Body surface area / day, and 100-500 μg of filgrastim or granulocyte colony-stimulating factor per day. Includes, The anti-CD33 targeting agent comprises HuM195 labeled with an effective amount of an alpha-emitting isotope, The cells expressing CD33 include blast cells, and The pharmaceutical composition is characterized in that it is for treating a hematological disease or disorder selected from the group consisting of multiple myeloma, acute myeloid leukemia, myelodysplastic syndrome, and myeloproliferative neoplasm.

2. The four individual chemotherapeutic agents mentioned above, 5 mg cladribine / 1 m of the subject 2 Body surface area / day, 2 g of cytarabine / 1 m of the subject 2 Body surface area / day, 10 mg of mitoxantrone / 1 m of the above subject 2 Body surface area / day, and 300 μg of filgrastim per day, The pharmaceutical composition according to claim 1, which is administered as such.

3. The four individual chemotherapeutic agents mentioned above, Cladribine was administered on days 1, 2, 3, 4, and 5 of the treatment period. Cytarabine was administered on days 1, 2, 3, 4, and 5 of the treatment period. Mitoxantrone was administered on the first, second, and third days of the treatment period, as well as The pharmaceutical composition according to claim 1 or 2, which is administered to the subject as filgrastim on day 0, day 1, day 2, day 3, day 4, and day 5 of the treatment period.

4. The pharmaceutical composition according to claim 1, wherein the anti-CD33 targeting agent is administered on any of the following days of the treatment period: from day 5 to day 30, from day 9 to day 20, or on day 14 of the treatment period.

5. The pharmaceutical composition according to claim 1, wherein the anti-CD33 targeting agent is administered in a protein dose of less than 16 mg / kg of body weight of the subject, or less than 10 mg / kg of body weight of the subject, or less than 6 mg / kg of body weight of the subject, or less than 50 μg / kg of body weight of the subject, or less than 20 μg / kg of body weight of the subject.

6. The aforementioned anti-CD33 targeting agent 225 The pharmaceutical composition according to claim 1, which is labeled with Ac and administered at a radiation dose of 0.7 to 0.8 μCi / kg of body weight of the subject, 0.8 to 0.9 μCi / kg of body weight of the subject, or 0.9 to 1.0 μCi / kg of body weight of the subject.

7. The pharmaceutical composition according to claim 1 for treating a hematological disorder or disorder selected from the group consisting of acute myeloid leukemia, myelodysplastic syndrome, and myeloproliferative neoplasm.

8. The aforementioned anti-CD33 targeting agent 225 The pharmaceutical composition according to claim 7, which is labeled with Ac and administered at a radiation dose of 0.7 to 0.8 μCi / kg of body weight of the subject, 0.8 to 0.9 μCi / kg of body weight of the subject, or 0.9 to 1.0 μCi / kg of body weight of the subject.

9. The pharmaceutical composition according to claim 8 for treating relapsed / refractory acute myeloid leukemia in the subject.

10. The anti-CD33 targeting agent is 225 Labeled with Ac and administered at a radiation dose of 0.7 to 0.8 μCi / kg of the body weight of the subject, 0.8 to 0.9 μCi / kg of the body weight of the subject, or 0.9 to 1.0 μCi / kg of the body weight of the subject. The pharmaceutical composition according to claim 9.

11. The pharmaceutical composition according to claim 1, wherein the anti-CD33 targeting agent is administered in a single dose.

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