Combination of antibody-drug conjugates and DNMT inhibitors

Abstract

A pharmaceutical product is provided for the administration of an antibody-drug conjugate in combination with a DNA methyltransferase (DNMT) inhibitor. The antibody-drug conjugate is an antibody-drug conjugate in which a drug-linker represented by the formula (wherein A represents a connection position to an antibody) is conjugated to the antibody, particularly an anti-TROP2 antibody, via a thioether bond. Also provided are therapeutic uses and methods in which an antibody-drug conjugate and a DNMT inhibitor are administered in combination to a subject: # imgabs0 #

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 428,945, filed on November 30, 2022, and U.S. Provisional Application No. 63 / 537,454, filed on September 8, 2023. For all purposes, each of the applications listed above is hereby incorporated by reference in its entirety. Technical field

[0003] The present disclosure relates to pharmaceutical products for the combination administration of a specific antibody - drug conjugate and a DNMT inhibitor, wherein the antibody - drug conjugate has an anti - tumor drug conjugated to an antibody, particularly an anti - TROP2 antibody, via a linker structure; and to therapeutic uses and methods in which the specific antibody - drug conjugate and the DNMT inhibitor are administered in combination to a subject.

[0004] Background

[0005] In the field of cancer immunotherapy, there is increasing recognition that DNA methyltransferase (DNMT) inhibitors are reagents that inhibit DNA methyltransferase enzymes, inducing DNA hypomethylation and growth inhibition or apoptosis in rapidly dividing cells. For example, decitabine ( 5 - aza - 2'- deoxycytidine) and azacitidine (Vidiza / 5 - aza - cytidine) are nucleoside analogs used to treat acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), and chronic myelomonocytic leukemia (CMML).

[0006] Antibody - drug conjugates (ADCs) consist of cytotoxic drugs conjugated to antibodies and are capable of selectively delivering the drug into cancer cells, resulting in cancer cell death (Ducry, L., et al., Bioconjugate Chem. (2010) 21, 5 - 13; Alley, S.C., et al., Current Opinion in Chemical Biology (2010) 14, 529 - 537; Damle N.K. Expert Opin. Biol. Ther. (2004) 4, 1445 - 1452; Senter P.D., et al., Nature Biotechnology (2012) 30, 631 - 637; Burris H.A., et al., J. Clin. Oncol. (2011) 29(4): 398 - 405).

[0007] One such antibody-drug conjugate is datopotamab deruxtecan (Dato-DXd, DS-1062a), which consists of a TROP2-targeting antibody and an exatecan derivative. Specifically, WO2015 / 098099 and WO2020 / 240467 provide a detailed description of exemplary TROP2-targeting antibody-drug conjugates including datopotamab deruxtecan. Datopotamab deruxtecan has shown clinical efficacy in multiple tumor types including lung cancer and breast cancer.

[0008] Inactivation of Schlafen11 (SLFN11) in cancer cells has been shown to confer resistance to anticancer agents that cause DNA damage and replication stress. Thus, SLFN11 can serve as a determinant of sensitivity to different types of DNA-damaging agents including, but not limited to, topoisomerase I inhibitors (Zoppoli et al., PNAS 2012; 109: 15030-35; Murai et al., Oncotarget 2016; 7: 76534-50; Murai et al., Mol. Cell 2018; 69: 371-84).

[0009] Zhao M, et al., AACR Cancer Res 2022; 82(12_Suppl): Abstract nr1791 reported that decitabine (a DNMT inhibitor) upregulates TROP2 and SLFN11 expression and enhances the antitumor efficacy of sacituzumab govitecan ( a humanized anti-TROP2 antibody conjugated to SN-38) in metaplastic carcinoma-derived xenograft cell lines and mesenchymal subtype breast cancer cell lines.

[0010] There remains a need for improved therapeutic compositions and methods that can enhance the efficacy of existing cancer therapeutics, increase the durability of the treatment response, improve patient tolerance, reduce dose-dependent toxicity, and / or provide alternative treatments for cancers that are resistant or refractory to previous cancer treatments. More specifically, there remains a need to identify further paired combinations with antibody-drug conjugates, particularly anti-TROP2 antibody-drug conjugates such as DS-1062a, to enhance their therapeutic potential. Accordingly, there is a need to provide drugs and treatment methods that can achieve excellent antitumor effects in cancer treatment, such as enhanced efficacy, increased durability of the treatment response, and / or reduced dose-dependent toxicity. SUMMARY OF THE INVENTION

[0011] The present disclosure provides a pharmaceutical product which exhibits excellent anti-tumor effects in cancer treatment by the combined administration of an antibody-drug conjugate, particularly an anti-TROP2 antibody-drug conjugate and a DNMT inhibitor. The present disclosure also provides a therapeutic use and method, in which the antibody-drug conjugate and the DNMT inhibitor are administered in combination to a subject.

[0012] Specifically, the present disclosure relates to the following [1] to

[27] :

[0013] [1] A pharmaceutical product comprising an antibody-drug conjugate and a DNMT inhibitor for combined administration, wherein the antibody-drug conjugate is an antibody-drug conjugate in which a drug-linker represented by the following formula is conjugated to an anti-TROP2 antibody via a thioether bond:

[0014]

[0015] wherein A represents the position of linkage to the antibody;

[0016] [2] The pharmaceutical product according to [1], wherein the anti-TROP2 antibody is an antibody comprising a heavy chain and a light chain, the heavy chain comprising CDRH1 consisting of the amino acid sequence shown in SEQ ID NO:3, CDRH2 consisting of the amino acid sequence shown in SEQ ID NO:4, and CDRH3 consisting of the amino acid sequence shown in SEQ ID NO:5, and the light chain comprising CDRL1 consisting of the amino acid sequence shown in SEQID NO:6, CDRL2 consisting of the amino acid sequence shown in SEQ ID NO:7, and CDRL3 consisting of the amino acid sequence shown in SEQ ID NO:8;

[0017] [3] The pharmaceutical product according to [2], wherein the anti-TROP2 antibody is an antibody comprising a heavy chain and a light chain, the heavy chain comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:9, and the light chain comprising a light chain variable region consisting of the amino acid sequence shown in SEQ IDNO:10;

[0018] [4] The pharmaceutical product according to [2] or [3], wherein the anti-TROP2 antibody is an antibody comprising a heavy chain and a light chain, the heavy chain consisting of the amino acid sequence shown in SEQ ID NO:12, and the light chain consisting of the amino acid sequence shown in SEQ ID NO:13;

[0019] [5] The pharmaceutical product according to [4], wherein the antibody lacks a lysine residue at the carboxyl terminus of the heavy chain;

[0020] [6] The pharmaceutical product according to any one of [1]-[5], wherein the average number of units of the drug-linker conjugated to each antibody molecule in the antibody-drug conjugate is in the range of 3.5-4.5;

[0021] [7] The pharmaceutical product according to any one of [1]-[5], wherein the antibody-drug conjugate is deruxtecan (DS-1062a);

[0022] [8] The pharmaceutical product according to any one of [1]-[7], wherein the DNMT inhibitor is decitabine or azacitidine, or a pharmaceutically acceptable salt thereof;

[0023] [9] The pharmaceutical product according to [8], wherein the DNMT inhibitor is decitabine or a pharmaceutically acceptable salt thereof;

[0024]

[10] The pharmaceutical product according to any one of [1]-[9], wherein the product is a composition, and the composition comprises an antibody-drug conjugate and a DNMT inhibitor for co-administration;

[0025]

[11] The pharmaceutical product according to any one of [1]-[9], wherein the product is a combination preparation, and the combination preparation comprises an antibody-drug conjugate and a DNMT inhibitor for sequential or separate co-administration;

[0026]

[12] The pharmaceutical product according to any one of [1]-

[11] , wherein the DNMT inhibitor is co-administered with a cytidine deaminase inhibitor;

[0027]

[13] The pharmaceutical product according to

[12] , wherein the cytidine deaminase inhibitor is cedazuridine or a pharmaceutically acceptable salt thereof;

[0028]

[14] The pharmaceutical product according to any one of [1]-

[13] , wherein the product is used for treating cancer;

[0029]

[15] The pharmaceutical product according to

[14] , wherein the cancer is at least one selected from the following: breast cancer, lung cancer, colorectal cancer, gastric cancer, esophageal cancer, head and neck cancer, gastroesophageal junction adenocarcinoma, biliary tract cancer, Paget's disease, pancreatic cancer, ovarian cancer, uterine carcinosarcoma, urothelial carcinoma, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumor, digestive tract stromal tumor, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, uterine body cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, glioblastoma multiforme, osteosarcoma, sarcoma and melanoma, cervical cancer, uterine cancer, testicular cancer and renal cell carcinoma;

[0030]

[16] The pharmaceutical product according to

[15] , wherein the cancer is colorectal cancer;

[0031]

[17] The pharmaceutical product according to

[15] , wherein the cancer is lung cancer;

[0032]

[18] The pharmaceutical product according to

[17] , wherein the lung cancer is non-small cell lung cancer;

[0033]

[19] The pharmaceutical product according to

[15] , wherein the cancer is lung cancer;

[0034]

[20] The pharmaceutical product according to any one of

[14] -

[19] , wherein the cancer cells of the cancer are SLFN11-deficient;

[0035]

[21] The pharmaceutical product according to

[20] , wherein the SLFN11 expression in the cancer cells of the patient is lower than that in the non-cancer cells expressing SLFN11 of the patient;

[0036]

[22] An antibody-drug conjugate for use in combination with a DNMT inhibitor in the treatment of cancer, wherein the antibody-drug conjugate and the DNMT inhibitor are as defined in any one of [1]-[9];

[0037]

[23] The antibody-drug conjugate for use according to

[22] , wherein the cancer is as defined in any one of

[15] -

[21] ;

[0038]

[24] The antibody-drug conjugate for use according to

[22] or

[23] , wherein the use comprises sequential administration of the antibody-drug conjugate and the DNMT inhibitor;

[0039]

[25] The antibody-drug conjugate for use according to

[22] or

[23] , wherein the use comprises separate and simultaneous administration of the antibody-drug conjugate and the DNMT inhibitor;

[0040]

[26] A method for treating cancer, which comprises co-administering an antibody-drug conjugate as defined in any one of [1]-[9] and a DNMT inhibitor to a subject in need thereof; and

[0041]

[27] The method according to

[26] , wherein the cancer is as defined in any one of

[15] -

[21] .

[0042] Advantages of the Invention

[0043] The present disclosure provides pharmaceutical products comprising a specific antibody-drug conjugate and a DNMT inhibitor for combination administration, wherein the antibody-drug conjugate has an anti-tumor drug conjugated to an antibody (particularly an anti-TROP2 antibody) via a linker structure; and provides therapeutic uses and methods, wherein the specific antibody-drug conjugate and the DNMT inhibitor are administered in combination to a subject. Accordingly, the present disclosure provides pharmaceuticals and treatments capable of achieving excellent anti-tumor effects in the treatment of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] [Anti-TROP2 antibody]:

[0045] Figure 1 is a diagram showing the amino acid sequence (SEQ ID NO:1) of the heavy chain of the anti-TROP2 antibody.

[0046] Figure 2 is a diagram showing the amino acid sequence (SEQ ID NO:2) of the light chain of the anti-TROP2 antibody.

[0047] Figure 3 is a diagram showing the amino acid sequence (SEQ ID NO:3 [= amino acid residues 50 - 54 of SEQ ID NO:1]) of heavy chain CDRH1.

[0048] Figure 4 is a diagram showing the amino acid sequence (SEQ ID NO:4 [= amino acid residues 69 - 85 of SEQ ID NO:1]) of heavy chain CDRH2.

[0049] Figure 5 is a diagram showing the amino acid sequence (SEQ ID NO:5 [= amino acid residues 118 - 129 of SEQ ID NO:1]) of heavy chain CDRH3.

[0050] Figure 6 is a diagram showing the amino acid sequence (SEQ ID NO:6 [= amino acid residues 44 - 54 of SEQ ID NO:2]) of light chain CDRL1.

[0051] Figure 7 is a diagram showing the amino acid sequence (SEQ ID NO:7 [= amino acid residues 70 - 76 of SEQ ID NO:2]) of light chain CDRL2.

[0052] Figure 8 is a diagram showing the amino acid sequence (SEQ ID NO:8 [= amino acid residues 109 - 117 of SEQ ID NO:2]) of light chain CDRL3.

[0053] Figure 9It is a figure showing the amino acid sequence of the heavy chain variable region (SEQ ID NO:9 [= amino acid residues 20 - 140 of SEQ ID NO:1]).

[0054] Figure 10 It is a figure showing the amino acid sequence of the light chain variable region (SEQ ID NO:10 [= amino acid residues 21 - 129 of SEQ ID NO:2]).

[0055] Figure 11 It is a figure showing the amino acid sequence of the heavy chain (SEQ ID NO:11 [= amino acid residues 20 - 469 of SEQ ID NO:1]).

[0056] [Experimental]:

[0057] Figure 12A and 12B It shows figures demonstrating the cell growth inhibitory activity of DS - 1062a in DLD - 1 cells and HCT - 15 cells, respectively, with or without pretreatment with decitabine (DAC).

[0058] Figure 13 It shows figures demonstrating the antitumor activity of DS - 1062a in a DLD - 1 xenograft mouse model with or without pretreatment with decitabine (DAC).

[0059] Figure 14 It shows figures demonstrating the cell growth inhibitory activity of DS - 1062a in DLD - 1 cells with or without pretreatment with azacitidine (AZA).

[0060] Figure 15 It shows figures demonstrating the antitumor activity of IMMU - 132 in a DLD - 1 xenograft mouse model with or without pretreatment with decitabine (DAC). Detailed Description of the Invention

[0062] To facilitate a better understanding of the present disclosure, certain terms are first defined. Additional definitions are set forth throughout the detailed description of the invention.

[0063] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised, 2000, Oxford University Press provide general dictionaries of many of the terms used in the art to one of ordinary skill in the art.

[0064] Unless the context requires otherwise, singular terms shall include the plural and plural terms shall include the singular.

[0065] Units, prefixes, and symbols are expressed in their accepted form of the International System of Units (SI). Numerical ranges include the numbers defining the range.

[0066] It should be understood that wherever aspects are described herein in the language “comprising,” other similar aspects described in “consisting of” and / or “consisting essentially of” are also provided.

[0067] The terms “inhibit” and “inhibition” can refer to a reduction of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of biological activity. Cell proliferation can be analyzed using techniques well known in the art, which measure the rate of cell division, and / or the fraction of cells in a cell population that undergo cell division, and / or the rate of loss of cells from a cell population due to terminal differentiation or cell death (e.g., thymidine incorporation).

[0068] The term “subject” refers to any animal (e.g., a mammal), including but not limited to humans, non-human primates, rodents, etc., that will be the recipient of a particular treatment. Generally, the terms “subject” and “patient” are used interchangeably herein when referring to a human subject.

[0069] The term "pharmaceutical product" means a preparation which is in a form permitting the biological activity of the active ingredient(s), as a composition containing all the active ingredients (for simultaneous administration), or as a combination of separate compositions each containing at least one but not all of the active ingredients (combination preparation) (for sequential or simultaneous administration), and which does not contain additional ingredients having unacceptable toxicity for the subject to whom the product is to be administered. Such a product may be sterile. "Simultaneous administration" means that the active ingredients are administered at the same time. "Sequential administration" means that the active ingredients are administered one after another in either order and with a time interval between the individual administrations. The time interval may be, for example, less than 24 hours, preferably less than 6 hours, more preferably less than 2 hours.

[0070] Terms such as "treat" or "treatment" or "manage" or "alleviate" or "mitigate" refer to both of the following: (1) therapeutic measures that cure, slow down, alleviate the symptoms of a diagnosed pathological condition or disorder and / or stop the progression of a diagnosed pathological condition or disorder; (2) preventive or prophylactic measures that prevent and / or slow down the development of a target pathological condition or disorder. Thus, those in need of treatment include: those who have already developed the disorder; those who are predisposed to developing the disorder; and those who need to prevent the disorder. In some aspects, if a patient shows, for example, a complete, partial or temporary remission of a certain type of cancer, the subject's cancer has been successfully "treated" according to the methods of the present disclosure.

[0071] The terms "cancer", "tumor", "cancerous" and "malignant" refer to or describe a physiological condition in mammals that is generally characterized by unregulated cell growth. Examples of cancers include, but are not limited to: breast cancer, lung cancer, colorectal cancer, gastric cancer, esophageal cancer, head and neck cancer, adenocarcinoma of the esophagogastric junction, biliary tract cancer, Paget's disease, pancreatic cancer, ovarian cancer, uterine carcinosarcoma, urothelial cancer, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumor, digestive tract stromal tumor, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, corpus cancer, renal cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer and renal cell carcinoma. Cancers include hematological malignancies and solid tumors, such as acute myeloid leukemia, multiple myeloma, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, Burkitt's lymphoma, follicular lymphoma; solid tumors such as breast cancer, lung cancer, neuroblastoma and colon cancer.

[0072] The term "cytotoxic agent" as used herein is defined broadly and refers to a substance that inhibits or interferes with cell function and / or causes cell destruction (cell death) and / or exerts an anti-tumor / anti-proliferative effect. For example, a cytotoxic agent directly or indirectly prevents the development, maturation, or spread of nascent tumor cells. The term also includes such agents that cause only cell growth inhibition and not just cytotoxicity. The term includes chemotherapeutic agents as specifically set forth below.

[0073] The term "chemotherapeutic agent" is a subset of the term "cytotoxic agent" and encompasses natural or synthetic chemical compounds.

[0074] In accordance with the methods or uses of the present disclosure, a compound of the present disclosure can be administered to a patient to facilitate a positive therapeutic response to cancer. The term "positive therapeutic response" with respect to cancer treatment refers to an improvement in symptoms associated with the disease. For example, an improvement in the disease can be characterized by a complete response. The term "complete response" means the absence of clinically detectable disease and normalization of any previous test results. Alternatively, an improvement in the disease can be classified as a partial response. "Positive therapeutic response" encompasses a reduction or inhibition in the progression and / or duration of cancer, a reduction or improvement in the severity of cancer, and / or an improvement in one or more of its symptoms caused by administration of a compound of the present disclosure. In certain aspects, such a term refers to one, two, or more than two results after administration of a compound of the present disclosure:

[0075] (1) Stabilization, reduction, or elimination of a population of cancer cells;

[0076] (2) Stabilization or reduction of cancer growth;

[0077] (3) Prevention of cancer formation;

[0078] (4) Eradication, removal, or control of primary, regional, and / or metastatic cancer;

[0079] (5) Reduction in mortality;

[0080] (6) Increase in disease-free, recurrence-free, progression-free, and / or overall survival period or survival rate;

[0081] (7) Increase in response rate, response durability, or number of responsive or remitting patients;

[0082] (8) Reduction in hospitalization rate;

[0083] (9) Reduction in length of hospitalization;

[0084] (10) The size of the cancer is maintained and does not increase or increases by less than 10%, preferably less than 5%, more preferably less than 4%, even more preferably less than 2%; and

[0085] (11) Increase in the number of patients in remission.

[0086] (12) Reduction in the number of adjuvant therapies (such as chemotherapy or hormone therapy) required for other cancer treatments.

[0087] Screening techniques can be used to assess clinical response, such as PET, magnetic resonance imaging (MRI) scans, X-ray imaging, computed tomography (CT) scans, flow cytometry or fluorescence-activated cell sorting (FACS) analysis, histology, gross pathology, and blood chemistry, including but not limited to changes detectable by ELISA, RIA, chromatography, etc. In addition to these positive treatment responses, subjects undergoing treatment can experience beneficial effects of improved symptoms related to the disease.

[0088] As used herein, the term "the expression level of SLFN11 is" a certain amount, such as 0%, means that the cancer cells of the said amount in the patient's cancer tissue express SLFN11. Similarly, as used herein, the term "the expression level of SLFN11 is <" a certain amount, such as 10%, means that less than the said amount of cancer cells in the patient's cancer tissue express SLFN11. The expression level of SLFN11 can be, for example, <25%, <20%, <15%, <10%, <9%, <8%, <7%, <6%, <5%, <4%, <3%, <2%, <1% or 0%.

[0089] As used herein, the term "deficient" means that the expression level of SLFN11 in a relevant patient, animal, tissue, cell, etc. is insufficient to exhibit a normal phenotype related to the gene, or insufficient to enable the protein to exhibit its physiological function. In the context of preclinical models, cells or animals in which the SLFN11 gene has been knocked out (KO) are examples of "SLFN11-deficient".

[0090] As used herein, the term "antibody" refers to a protein capable of recognizing and specifically binding to an antigen. A normal or conventional mammalian antibody comprises a tetramer, which typically consists of two pairs of identical polypeptide chains, each pair consisting of one "light" chain (usually having a molecular weight of about 25 kDa) and one "heavy" chain (usually having a molecular weight of about 50 - 70 kDa). As used herein, the terms "heavy chain" and "light chain" refer to any immunoglobulin polypeptide having sufficient variable domain sequence to confer specificity for a target antigen. The amino-terminal portion of each light and heavy chain typically comprises a variable domain of about 100 - 110 or more amino acids, which is generally responsible for antigen recognition. As used herein, the terms "variable region" or "variable domain" are used interchangeably and are common in the art. The carboxyl-terminal portion of each chain is typically defined as the constant domain responsible for effector function. Thus, in a naturally occurring antibody, the full-length heavy-chain immunoglobulin polypeptide comprises a variable domain (V H ) and three constant domains (C H1 , C H2 and C H3 ) and a hinge region between C H1 and C H2 , wherein the VH domain is located at the amino terminus of the polypeptide and the C H3 domain is located at the carboxyl terminus, and the full-length light-chain immunoglobulin polypeptide comprises a variable domain (V L ) and a constant domain (C L ), wherein the V L domain is located at the amino terminus of the polypeptide and the C L domain is located at the carboxyl terminus. However, those skilled in the art will understand that the positions of the domains in a naturally occurring antibody can be modified in certain antibody-like binding proteins without loss of antigen-binding ability. The classes of human light chains are called kappa and lambda light chains.

[0091] Within the full-length light and heavy chains, the variable and constant domains are typically joined by a "J" region of about 12 or more amino acids, and the heavy chain also includes a "D" region of about 10 or more amino acids. The variable regions of each light / heavy chain pair typically form the antigen-binding site. The variable domains of naturally occurring antibodies generally exhibit the same general structure as relatively conserved framework regions (FRs) connected by three hypervariable regions (also called complementarity-determining regions or CDRs). The CDRs from each pair of two chains are typically aligned by the framework regions, which may enable binding to a specific epitope. From the amino terminus to the carboxyl terminus, the light and heavy chain variable domains generally each contain the domains FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0092] The term "antibody fragment" refers to an intact or full-length chain or part of an antibody, typically the target binding region or variable region. Examples of antibody fragments include, but are not limited to: F ab -, F ab' -, F (ab')2 - and F v -fragments. As used herein, the term "functional fragment" is generally synonymous with "antibody fragment", and for antibodies, antibody fragments such as F v -, F ab -, F (ab')2 - may be mentioned.

[0093] The numbering of the amino acid residues referred to herein is according to the EU numbering system (also described in Kabat et al, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)).

[0094] A "monoclonal" antibody or an antigen-binding fragment thereof refers to a population of homologous antibodies or antigen-binding fragments that exhibit highly specific binding to a single antigenic determinant or epitope. This is in contrast to polyclonal antibodies, which typically include different antibodies directed against different antigenic determinants. The term "monoclonal" antibody or an antigen-binding fragment thereof encompasses intact and full-length monoclonal antibodies as well as antibody fragments (such as Fab, Fab’, F(ab’)2, Fv), single-chain (scFv) mutants, fusion proteins comprising antibody portions, and any other modified immunoglobulin molecules comprising an antigen recognition site. In addition, a "monoclonal" antibody or an antigen-binding fragment thereof refers to such antibodies and their antigen-binding fragments produced by means including, but not limited to, hybridoma, phage selection, recombinant expression, and transgenic animals.

[0095] As used herein, the term "antigen" or "target antigen" refers to a molecule or part of a molecule that can be recognized and bound by a binding protein disclosed herein. This target antigen can be used in animals to produce antibodies that can bind to the epitopes of the antigen. The target antigen can have one or more epitopes.

[0096] As used herein, the term "epitope" refers to the region or structural element of an antigen that is recognized and bound by a binding protein of the present disclosure. More precisely, an epitope is a specific structure bound by the CDR of a binding protein. An epitope can include protein structural elements, carbohydrates, or even portions of lipid structures found in membranes. When a binding protein preferentially recognizes its antigen target in a complex mixture of proteins and / or macromolecules, the binding protein is said to specifically bind to the antigen. The term "specifically binds" refers to a binding protein that specifically binds to a specific binding molecule or a fragment thereof (such as an antigen). As determined by, for example, immunoassays, BIAcore, or other assays known in the art, a binding protein of a specific binding molecule or a fragment thereof can bind to other molecules with a lower affinity. Specifically, an antibody or fragment that specifically binds to at least one molecule or a fragment thereof can compete off non-specifically bound molecules.

[0097] As used herein, the term "antigen-binding site" refers to the site created on the surface of a binding protein of the present disclosure to which an antigen or an epitope on the antigen binds. The antigen-binding site of a binding protein is typically described with reference to the loop structures created by the complementarity-determining regions (CDRs) of the binding protein. Detailed Description

[0098] Hereinafter, preferred modes for implementing the present disclosure will be described. The embodiments described below are merely examples of typical embodiments of the present disclosure and are not intended to limit the scope of the present disclosure.

[0099] 1. Antibody-Drug Conjugates

[0100] The antibody-drug conjugates used in the present disclosure are the following antibody-drug conjugates, in which the drug-linker represented by the following formula is conjugated to an antibody, particularly an anti-TROP2 antibody, via a thioether bond:

[0101]

[0102] Wherein A represents the position of attachment to the antibody.

[0103] In the present disclosure, the partial structure composed of the linker and the drug in the antibody-drug conjugate is referred to as the "drug-linker". This drug-linker is attached to the thiol group (in other words, the sulfur atom of the cysteine residue) at the interchain disulfide bond sites formed in the antibody (two sites between the heavy chains and two sites between the heavy chain and the light chain).

[0104] The drug-linker of the present disclosure includes irinotecan (IUPAC name: (1S,9S)-1-amino-9-ethyl-5-fluoro-1,2,3,9,12,15-hexahydro-9-hydroxy-4-methyl-10H,13H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13-dione (also represented as the chemical name: (1S,9S)-1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13(9H,15H)-dione)) as a component, which is a topoisomerase I inhibitor. Irinotecan is a camptothecin derivative with antitumor effects and is represented by the following formula:

[0105]

[0106] The antibody-drug conjugate used in the present disclosure can also be represented by the following formula:

[0107]

[0108] Here, the drug-linker is conjugated to the antibody ("Antibody-") via a thioether bond, especially an anti-TROP2 antibody. The meaning of n is the same as the so-called average number of conjugated drug molecules (DAR; Drug-to-Antibody Ratio), which represents the average number of units of the drug-linker conjugated to each antibody molecule.

[0109] After migrating into cancer cells, the antibody-drug conjugate used in the present disclosure is cleaved at the linker portion to release a compound represented by the following formula:

[0110]

[0111] 2. Antibody in the Antibody-Drug Conjugate

[0112] The antibody in the antibody-drug conjugate used in the present disclosure is an anti-TROP2 antibody and can be derived from any species, preferably from humans, rats, mice or rabbits. When the antibody is derived from a species other than the human species, chimerization or humanization is preferably performed using well-known techniques. The antibody can be a polyclonal antibody or a monoclonal antibody, and is preferably a monoclonal antibody.

[0113] The antibody in the antibody-drug conjugate used in the present disclosure is preferably an antibody having the property of being able to target cancer cells, and is preferably an antibody having the following properties, such as the property of recognizing cancer cells, the property of binding to cancer cells, the property of internalizing into cancer cells and / or the cytotoxic activity against cancer cells.

[0114] The binding activity of the antibody to cancer cells can be confirmed by flow cytometry. The following assays can be used to confirm the internalization of the antibody into cancer cells: (1) an assay for visualizing the antibody integrated in cells under a fluorescence microscope using a secondary antibody (fluorescently labeled) that binds to the therapeutic antibody (Cell Death and Differentiation (2008) 15, 751-761); (2) an assay for measuring the fluorescence intensity integrated in cells using a secondary antibody (fluorescently labeled) that binds to the therapeutic antibody (Molecular Biology of the Cell, Vol. 15, 5268-5282, December 2004); or (3) the Mab-ZAP assay using an immunotoxin that binds to the therapeutic antibody, wherein the toxin is released after integration into cells to inhibit cell growth (Bio Techniques 28:162-165, January 2000). As the immunotoxin, a recombinant complex protein of the catalytic domain of diphtheria toxin and protein G can be used.

[0115] The anti-tumor activity of the antibody can be confirmed by in vitro assays for inhibitory activity against cell growth. For example, a cancer cell line overexpressing the target protein for the antibody is cultured, and the antibody is added to the culture system at different concentrations to determine the inhibitory activity against lesion formation, colony formation, and spheroid growth. The anti-tumor activity can be confirmed in vivo, for example, by administering the antibody to nude mice transplanted with a cancer cell line highly expressing the target protein and measuring the changes in the cancer cells.

[0116] Since the conjugated compound in the antibody-drug conjugate exerts an anti-tumor effect, it is preferred but not necessary that the antibody itself should have an anti-tumor effect. In order to specifically and selectively exert the cytotoxic activity of the anti-tumor compound against cancer cells, it is important and preferred that the antibody should have the property of internalizing to migrate into cancer cells.

[0117] The anti-TROP2 antibody in the antibody-drug conjugate used in the present disclosure can be obtained by procedures known in the art. For example, the antibodies of the present disclosure can be obtained using methods commonly practiced in the art, which involve immunizing an animal with an antigenic polypeptide and collecting and purifying the antibody produced in vivo. The source of the antigen is not limited to humans, and animals can be immunized with antigens derived from non-human animals such as mice and rats. In such cases, the cross-reactivity of the antibody binding to the obtained heterologous antigen with the human antigen can be tested to screen antibodies applicable to human diseases.

[0118] Alternatively, according to methods known in the art (e.g., Kohler and Milstein, Nature (1975) 256, p. 495-497; and Kennet, R., ed., Monoclonal Antibodies, p. 365-367, Plenum Press, N.Y. (1980)), antibody-producing cells that produce antibodies against an antigen are fused with myeloma cells to establish hybridomas, from which monoclonal antibodies can be obtained.

[0119] The antigen can be obtained by genetically engineering a host cell to produce a gene encoding an antigenic protein. Specifically, a vector that allows expression of the antigen gene is prepared and transferred into the host cell to express the gene. The antigen thus expressed can be purified. Antibodies can also be obtained by immunizing an animal with the above-described genetically engineered antigen-expressing cells or cell lines expressing the antigen.

[0120] The anti-TROP2 antibody in the antibody-drug conjugate used in the present disclosure is preferably a recombinant antibody obtained by artificial modification for the purpose of reducing heterologous antigenicity to humans, such as a chimeric antibody or a humanized antibody; or preferably an antibody having only antibody gene sequences derived from humans, i.e., a human antibody. These antibodies can be manufactured using known methods.

[0121] As a chimeric antibody, an antibody can be exemplified in which the antibody variable region and constant region are derived from different species. For example, a chimeric antibody in which the antibody variable region derived from a mouse or rat is linked to the antibody constant region derived from a human (Proc. Natl. Acad. Sci. USA, 81, 6851-6855, (1984)).

[0122] As a humanized antibody, an antibody can be exemplified that is obtained by integrating only the complementarity-determining regions (CDRs) of a heterologous antibody into an antibody derived from a human (Nature (1986) 321, pp. 522-525), an antibody obtained by grafting a portion of the amino acid residues of the heterologous antibody framework and the CDR sequence of the heterologous antibody onto a human antibody by the CDR-grafting method (WO90 / 07861), and an antibody humanized using a gene conversion mutagenesis strategy (U.S. Patent No. 5,821,337).

[0123] As a human antibody, antibodies produced by mice that produce human antibodies using a human chromosome fragment (the fragment includes the heavy and light chain genes of a human antibody) can be exemplified (see Tomizuka, K. et al., Nature Genetics (1997) 16, p. 133-143; Kuroiwa, Y. et.al., Nucl. Acids Res. (1998) 26, p. 3447-3448; Yoshida, H. et.al., Animal Cell Technology: Basic and Applied Aspects vol. 10, p. 69-73 (Kitagawa, Y., Matsuda, T. and Iijima, S. eds.), Kluwer Academic Publishers, 1999; Tomizuka, K. et.al., Proc. Natl. Acad. Sci. USA (2000) 97, p. 722-727, etc.). As an alternative, antibodies obtained by phage display and selected from a human antibody library can be exemplified (see Wormstone, I.M. et.al, Investigative Ophthalmology & Visual Science. (2002) 43(7), p. 2301-2308; Carmen, S. et.al., Briefings in Functional Genomics and Proteomics (2002), 1(2), p. 189-203; Siriwardena, D. et.al., Ophthalmology (2002) 109(3), p. 427-431, etc.).

[0124] Among the antibodies in the antibody-drug conjugates used in the present disclosure, also included are modified variants of the antibody. The modified variant refers to a variant obtained by chemically or biologically modifying the antibody described in the present disclosure. Examples of chemically modified variants include the following variants: variants including the linkage of a chemical moiety to the amino acid backbone, variants including the linkage of a chemical moiety to an N-linked or O-linked carbohydrate chain, etc. Examples of biologically modified variants include: variants obtained by post-translational modification (such as N-linked or O-linked glycosylation, N- or C-terminal processing, deamidation, isomerization of aspartic acid, or oxidation of methionine), and variants that have had a methionine residue added to the N-terminus by expression in a prokaryotic host cell. Further, antibodies labeled to achieve detection or separation of the antibody or antigen described in the present disclosure, such as enzyme-labeled antibodies, fluorescent-labeled antibodies, and affinity-labeled antibodies are also included within the meaning of the modified variant. Such modified variants of the antibody described in the present disclosure are useful for improving the stability and blood retention of the antibody, reducing its antigenicity, detecting or separating the antibody or antigen, etc.

[0125] In addition, by regulating the modification of glycans (glycosylation, defucosylation, etc.) linked to the antibody described in the present disclosure, antibody-dependent cell cytotoxic activity can be enhanced. As techniques for regulating the glycan modification of an antibody, those disclosed in WO99 / 54342, WO00 / 61739, WO02 / 31140, WO2007 / 133855, WO2013 / 120066, etc. are known. However, the technique is not limited thereto. Among the anti-TROP2 antibodies described in the present disclosure, also included are antibodies in which the glycan modification is regulated.

[0126] It is known that lysine residues at the carboxyl terminus of the heavy chain of antibodies produced in cultured mammalian cells are deleted (Journal of Chromatography A, 705: 129 - 134 (1995)). It is also known that two amino acid residues (glycine and lysine) at the carboxyl terminus of the heavy chain of antibodies produced in cultured mammalian cells are deleted, and the newly located proline residue at the carboxyl terminus is amidated (Analytical Biochemistry, 360: 75 - 83 (2007)). However, such deletions and modifications of the heavy chain sequence do not affect the antigen - binding affinity and effector functions (activation of complement, antibody - dependent cytotoxicity, etc.) of the antibody. Therefore, among the anti - TROP2 antibodies described in the present disclosure, there are also included antibodies and functional fragments of antibodies that have undergone such modifications, and also deletion variants in which one or two amino acids are deleted at the carboxyl terminus of the heavy chain, variants obtained by amidating the deletion variants (for example, heavy chains in which the carboxyl - terminal proline residue has been amidated), etc. The types of deletion variants with deletions at the carboxyl terminus of the anti - body heavy chain described in the present disclosure are not limited to the above - mentioned variants, as long as the antigen - binding affinity and effector functions are retained. The two heavy chains constituting the antibody described in the present disclosure can be of one type selected from the group consisting of full - length heavy chains and the above - mentioned deletion variants, or can be a combination of two types selected therefrom. The proportional amount of each deletion variant can be affected by the type and culture conditions of the cultured mammalian cells that produce the antibody described in the present disclosure; however, preferably, antibodies in which one amino acid residue at the carboxyl terminus is deleted in both of the two heavy chains of the antibody described in the present disclosure can be exemplified.

[0127] As isotypes of the anti - TROP2 antibodies described in the present disclosure, for example, IgG (IgG1, IgG2, IgG3, IgG4) can be exemplified, and preferably IgG1 can be exemplified.

[0128] In the present disclosure, the term "anti - TROP2 antibody" refers to an antibody that specifically binds to TROP2 (TACSTD2: tumor - associated calcium signal transducer 2; EGP - 1), and preferably has the activity of internalizing into TROP2 - expressing cells by binding to TROP2.

[0129] Examples of anti - TROP2 antibodies include hTINA1 - H1L1 (WO2015 / 098099), and preferably datopotamab can be exemplified.

[0130] 3. Manufacture of Antibody - Drug Conjugates

[0131] The drug - linker intermediate for manufacturing the antibody - drug conjugate described in the present disclosure is represented by the following formula:

[0132]

[0133] The drug-linker intermediate can be represented by the chemical name N-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl]glycylglycyl-L-phenylalanyl-N-[(2-{[(1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinolin-1-yl]amino}-2-oxoethoxy)methyl]glycinamide, and can be manufactured with reference to the descriptions in WO2014 / 057687, WO2015 / 098099, WO2019 / 044947, etc.

[0134] The antibody-drug conjugate used in the present disclosure can be manufactured by reacting the above drug-linker intermediate with an anti-TROP2 antibody having a thiol group (also referred to as a sulfhydryl group).

[0135] The anti-TROP2 antibody having a thiol group can be obtained by methods well known in the art (Hermanson, G.T, Bioconjugate Techniques, pp.56 - 136, pp.456 - 493, Academic Press (1996)). For example, by using a reducing agent with 0.3 - 3 molar equivalents of inter-chain disulfide bonds per chain in the antibody, such as tris(2-carboxyethyl)phosphine hydrochloride (TCEP), and reacting with the antibody in a buffer containing a chelating agent such as ethylenediaminetetraacetic acid (EDTA), an anti-TROP2 antibody having a thiol group can be obtained, and the inter-chain disulfide bonds in the antibody are partially or completely reduced.

[0136] Furthermore, by using 2 - 20 molar equivalents of the drug-linker intermediate relative to the anti-TROP2 antibody having a thiol group, an antibody-drug conjugate conjugated with 2 - 8 drug molecules per antibody molecule can be manufactured.

[0137] The average number of drug molecules conjugated to each anti-TROP2 antibody molecule of the manufactured antibody-drug conjugate can be measured, for example, by the following methods: a calculation method based on the measurement of the UV absorbance of the antibody-drug conjugate and its conjugation precursor at two wavelengths of 280 nm and 370 nm (UV method); or a calculation method based on the quantification measured by HPLC of the fragment obtained by treating the antibody-drug conjugate with a reducing agent (HPLC method).

[0138] The conjugation between the anti-TROP2 antibody and the drug-linker intermediate and the calculation of the average number of drug molecules conjugated per antibody molecule of the antibody-drug conjugate can be carried out with reference to the descriptions in WO2014 / 057687, WO2015 / 098099, WO2017 / 002776, WO2022 / 014698, etc.

[0139] In the present disclosure, the term "anti-TROP2 antibody-drug conjugate" refers to an antibody-drug conjugate in which the antibody in the antibody-drug conjugate described in the present disclosure is an anti-TROP2 antibody.

[0140] The anti-TROP2 antibody is preferably an antibody comprising a heavy chain and a light chain, the heavy chain comprising CDRH1 [= the amino acid sequence consisting of amino acid residues 50-54 of SEQ ID NO:1] consisting of the amino acid sequence shown in SEQ ID NO:3, CDRH2 [= the amino acid sequence consisting of amino acid residues 69-85 of SEQ ID NO:1] consisting of the amino acid sequence shown in SEQ ID NO:4, and CDRH3 [= the amino acid sequence consisting of amino acid residues 118-129 of SEQ ID NO:1] consisting of the amino acid sequence shown in SEQ ID NO:5, and the light chain comprising CDRL1 [= the amino acid sequence consisting of amino acid residues 44-54 of SEQ ID NO:2] consisting of the amino acid sequence shown in SEQ ID NO:6, CDRL2 [= the amino acid sequence consisting of amino acid residues 70-76 of SEQ ID NO:2] consisting of the amino acid sequence shown in SEQ ID NO:7, and CDRL3 [= the amino acid sequence consisting of amino acid residues 109-117 of SEQ ID NO:2] consisting of the amino acid sequence shown in SEQ ID NO:8;

[0141] More preferably, it is an antibody comprising a heavy chain and a light chain, the heavy chain comprising a heavy chain variable region [= the amino acid sequence consisting of amino acid residues 20-140 of SEQ ID NO:1] consisting of the amino acid sequence shown in SEQ ID NO:9, and the light chain comprising a light chain variable region [= the amino acid sequence consisting of amino acid residues 21-129 of SEQ ID NO:2] consisting of the amino acid sequence shown in SEQ ID NO:10; and

[0142] Even more preferably, it is an antibody comprising a heavy chain and a light chain, wherein the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 12 [= the amino acid sequence consisting of amino acid residues 20-470 of SEQ ID NO: 1], and the light chain consists of the amino acid sequence shown in SEQ ID NO: 13 [= amino acid residues 21-234 of SEQ ID NO: 2]; or an antibody comprising a heavy chain and a light chain, wherein the heavy chain consists of the amino acid sequence shown in SEQ ID NO: 11 [= the amino acid sequence consisting of amino acid residues 20-469 of SEQ ID NO: 1], and the light chain consists of the amino acid sequence shown in SEQ ID NO: 13 [= amino acid residues 21-234 of SEQ ID NO: 2].

[0143] In the anti-TROP2 antibody-drug conjugate, the average number of drug-linker units conjugated per molecule of antibody is preferably 2-8, more preferably 3-5, even more preferably 3.5-4.5, and even more preferably about 4.

[0144] The anti-TROP2 antibody-drug conjugate can be manufactured with reference to the descriptions in WO2015 / 098099, WO2017 / 002776, and WO2022 / 014698.

[0145] In a preferred embodiment, the anti-TROP2 antibody-drug conjugate is datopotamab deruxtecan (DS-1062a).

[0146] 4. DNA methyltransferase (DNMT) inhibitors

[0147] In the present disclosure, the terms "DNA methyltransferase inhibitor" and "DNMT inhibitor" refer to compounds that inhibit DNA methyltransferase enzymes. In some aspects, the DNMT inhibitor inhibits one or more of human DNA methyltransferase enzymes, including DNMT1, DNMT2, DNMT3a, and DNMT3b.

[0148] In some aspects, the DNA methyltransferase inhibitor is selected from decitabine (5-aza-2'-deoxycytidine), azacitidine (5-azacytidine), guadecitabine, 5,6-dihydro-5-azacytidine, fazarabine, 5-fluoro-2'-deoxycytidine, zebularine, hydralizine, procaine, procainamide, epigallocatechin gallate, palmapurine A, (S)-2-(1,3-dioxo-1,3-dihydroisoindol-2-yl)-3-(1H-indol-3-yl)-propanoic acid, and pharmaceutically acceptable salts thereof. Preferred DNA methyltransferase inhibitors include: decitabine (5-aza-2'-deoxycytidine), azacitidine (5-azacytidine), 5,6-dihydro-5-azacytidine, fazarabine, 5-fluoro-2'-deoxycytidine, zebularine, and pharmaceutically acceptable salts thereof. Preferably, the DNMT inhibitor is decitabine or azacitidine, or a pharmaceutically acceptable salt thereof, particularly decitabine or a pharmaceutically acceptable salt thereof.

[0149] 5. Combination of an antibody-drug conjugate and a DNMT inhibitor

[0150] In the combination embodiments of the present disclosure, the antibody-drug conjugate combined with the DNMT inhibitor is an antibody-drug conjugate in which the antibody is an anti-TROP2 antibody.

[0151] In an embodiment of the above-described combination embodiment, the anti-TROP2 antibody comprises a heavy chain and a light chain. The heavy chain comprises CDRH1 consisting of the amino acid sequence shown in SEQ ID NO:3 [= amino acid residues 50-54 of SEQ ID NO:1], CDRH2 consisting of the amino acid sequence shown in SEQ ID NO:4 [= amino acid residues 69-85 of SEQ ID NO:1], and CDRH3 consisting of the amino acid sequence shown in SEQ ID NO:5 [= amino acid residues 118-129 of SEQ ID NO:1]. The light chain comprises CDRL1 consisting of the amino acid sequence shown in SEQ ID NO:6 [= amino acid residues 44-54 of SEQ ID NO:2], CDRL2 consisting of the amino acid sequence shown in SEQ ID NO:7 [= amino acid residues 70-76 of SEQ ID NO:2], and CDRL3 consisting of the amino acid sequence shown in SEQ ID NO:8 [= amino acid residues 109-117 of SEQ ID NO:2]. In another embodiment of the above-described combination embodiment, the anti-TROP2 antibody comprises a heavy chain and a light chain. The heavy chain comprises a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:9 [= amino acid residues 20-140 of SEQ ID NO:1], and the light chain comprises a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:10 [= amino acid residues 21-129 of SEQ ID NO:2]. In another embodiment of the above-described combination embodiment, the anti-TROP2 antibody comprises a heavy chain and a light chain. The heavy chain consists of the amino acid sequence shown in SEQ ID NO:12 [= amino acid residues 20-470 of SEQ ID NO:1], and the light chain consists of the amino acid sequence shown in SEQ ID NO:13 [= amino acid residues 21-234 of SEQ ID NO:2]. In another embodiment of the above-described combination embodiment, the anti-TROP2 antibody comprises a heavy chain and a light chain. The heavy chain consists of the amino acid sequence shown in SEQ ID NO:11 [= amino acid residues 20-469 of SEQ ID NO:1], and the light chain consists of the amino acid sequence shown in SEQ ID NO:13 [= amino acid residues 21-234 of SEQ ID NO:2]. In another embodiment of the above-described combination embodiment, the anti-TROP2 antibody is datopotamab.

[0152] In a particularly preferred embodiment of the above-described combination embodiment, the anti-TROP2 antibody-drug conjugate is datopotamab deruxtecan (DS-1062a), and the DNMT inhibitor is decitabine.

[0153] In a particularly preferred embodiment of the above combination embodiment, the anti-TROP2 antibody-drug conjugate is datopotamab deruxtecan (DS-1062a), and the DNMT inhibitor is azacitidine.

[0154] In some aspects of the above combination embodiment, the antibody-drug conjugate and / or the DNMT inhibitor are further administered in combination with one or more chemotherapeutic agents. In some aspects, the DNMT inhibitor is further administered in combination with a cytidine deaminase (CDA) inhibitor, preferably in combination with cedazuridine or tetrahydrouridine, particularly preferably in combination with cedazuridine, preferably as a fixed-dose combination for oral administration such as ASTX727 for oral administration ( decitabine / cedazuridine).

[0155] 6. Therapeutic combination uses and methods

[0156] Described below are pharmaceutical products and therapeutic uses and methods in which the anti-TROP2 antibody-drug conjugate described in the present disclosure is administered in combination with a DNMT inhibitor.

[0157] The pharmaceutical products and therapeutic uses and methods of the present disclosure may be characterized in that the antibody-drug conjugate and the DNMT inhibitor are included as active ingredients in different formulations and are administered simultaneously or at different times, or may be characterized in that the antibody-drug conjugate and the DNMT inhibitor are included as active ingredients in a single formulation and are administered.

[0158] In the pharmaceutical products and therapeutic methods of the present disclosure, the single DNMT inhibitor used in the present disclosure may be administered in combination with the antibody-drug conjugate, or two or more different DNMT inhibitors may be administered in combination with the antibody-drug conjugate.

[0159] The pharmaceutical products and therapeutic methods of the present disclosure can be used for the treatment of cancer, and can preferably be used for the treatment of at least 1 cancer selected from the following: breast cancer, lung cancer, colorectal cancer, gastric cancer, esophageal cancer, head and neck cancer, adenocarcinoma of the esophagogastric junction, biliary tract cancer, Paget's disease, pancreatic cancer, ovarian cancer, uterine carcinosarcoma, urothelial cancer, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumor, digestive tract stromal tumor, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, uterine body cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer and renal cell carcinoma.

[0160] The presence or absence of a tumor marker such as the TROP2 tumor marker can be determined, for example, by collecting tumor tissue from a cancer patient to prepare a formalin-fixed, paraffin-embedded (FFPE) sample and performing a gene product (protein) test on the sample using, for example, immunohistochemistry (IHC), flow cytometry, or western blotting; or performing a gene transcription test using, for example, in situ hybridization (ISH), quantitative PCR (q-PCR), or microarray analysis; or by collecting cell-free circulating tumor DNA (ctDNA) from a cancer patient and testing the ctDNA using a method such as next-generation sequencing (NGS).

[0161] The pharmaceutical products and treatment methods of the present disclosure can be preferably used for mammals, but more preferably for humans.

[0162] The anti-tumor effects of the pharmaceutical products and treatment methods of the present disclosure can be confirmed by transplanting cancer cells into test animals to prepare a model and measuring the tumor volume reduction or life extension effects caused by administering the pharmaceutical products and treatment methods of the present disclosure. Then, the effects of the combined use of the antibody-drug conjugates and DNMT inhibitors used in the present disclosure can be confirmed by comparing the anti-tumor effects of the antibody-drug conjugates and the DNMT inhibitors administered alone used in the present disclosure.

[0163] The anti-tumor effects of the pharmaceutical products and treatment methods of the present disclosure can be confirmed in clinical trials using any evaluation method that utilizes the Response Evaluation Criteria in Solid Tumors (RECIST), WHO evaluation method, Macdonald evaluation method, body weight measurement, and other methods, and can be determined based on indicators such as complete response (CR), partial response (PR); progressive disease (PD), objective response rate (ORR), duration of response (DoR), progression-free survival (PFS), overall survival (OS), etc.

[0164] By using the above methods, the superiority of the anti-tumor effects of the pharmaceutical products and treatment methods of the present disclosure over existing pharmaceutical products and treatment methods for cancer treatment can be confirmed.

[0165] The pharmaceutical products and treatment methods of the present disclosure can delay the development of cancer cells, inhibit their growth, and further kill cancer cells. These effects can relieve cancer patients of the symptoms caused by cancer, or achieve an improvement in the quality of life (QOL) of cancer patients, and achieve a therapeutic effect by maintaining the life of cancer patients. Even if the pharmaceutical products and treatment methods of the present disclosure cannot completely kill cancer cells, they can achieve a higher QOL for cancer patients while achieving longer-term survival by inhibiting or controlling the growth of cancer cells.

[0166] It is expected that the pharmaceutical products of the present disclosure exert a therapeutic effect by administering to a patient as a systemic treatment or by locally administering to cancer tissues.

[0167] On the other hand, the pharmaceutical products and treatment methods of the present disclosure provide for use as an adjuvant in cancer treatment using ionizing radiation or other chemotherapeutic agents. For example, in the treatment of cancer, the treatment may comprise administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical product, either concomitantly or sequentially with ionizing radiation or other chemotherapeutic agents.

[0168] The pharmaceutical products and treatment methods of the present disclosure can be used as adjuvant chemotherapy in combination with surgery. The pharmaceutical products of the present disclosure can be administered before surgery for the purpose of reducing the tumor size (referred to as neoadjuvant chemotherapy or neoadjuvant treatment), or can be administered after surgery for the purpose of preventing tumor recurrence (referred to as adjuvant chemotherapy or adjuvant therapy).

[0169] In some embodiments, cancer cells can have a BRCA1 and / or BRCA2-deficient phenotype, i.e., reduced or absent BRCA1 and / or BRCA2 activity in the cancer cells. Cancer cells with such a phenotype may lack BRCA1 and / or BRCA2, i.e., the expression and / or activity of BRCA1 and / or BRCA2 in the cancer cells may be reduced or absent, for example, by mutations or polymorphisms in the encoding nucleic acid, or by amplification, mutation or polymorphism of a gene encoding a regulator, such as the EMSY gene encoding a BRCA2 regulator (Hughes-Davies, et al., Cell, 115, 523-535). BRCA1 and BRCA2 are known tumor suppressors, and their wild-type alleles are often lost in tumors of heterozygous carriers (Jasin M., Oncogene, 21(58), 8981-93(2002); Tutt, et al., Trends Mol Med., 8(12), 571-6, (2002)). The association of BRCA1 and / or BRCA2 mutations with breast cancer has been well characterized in the art (Radice, P.J., Exp Clin Cancer Res., 21(3 Suppl), 9-12(2002)). Amplification of the EMSY gene encoding a BRCA2-binding factor is also associated with breast and ovarian cancer. Carriers of BRCA1 and / or BRCA2 mutations also have a higher risk of developing certain cancers, including breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer, and lung cancer. In some embodiments, an individual is heterozygous for one or more variants (e.g., mutations and polymorphisms) in BRCA1 and / or BRCA2 or their regulators. Detection of variants in BRCA1 and BRCA2 is well known in the art and is described, for example, in EP 699754, EP 705903, Neuhausen, S.L. and Ostrander, E.A., Genet. Test, 1, 75-83(1992); Chappnis, P.O. and Foulkes, W.O., Cancer Treat Res, 107, 29-59(2002); Janatova M., et al., Neoplasma, 50(4), 246-505(2003); Jancarkova, N., Ceska Gynekol., 68{1), 11-6(2003)). The determination of EMSY amplification of BRCA2-binding factor is described in Hughes-Davies, et al., Cell, 115, 523-535).

[0170] Cancer-related mutations and polymorphisms can be detected at the nucleic acid level by detecting the presence of variant nucleic acid sequences, or at the protein level by detecting the presence of variant (i.e., mutant or allelic variant) polypeptides.

[0171] The pharmaceutical products of the present disclosure can be administered in the presence of at least one pharmaceutically suitable ingredient. Depending on the dosage, administration concentration, etc. of the antibody-drug conjugate and the DNMT inhibitor used in the present disclosure, pharmaceutically suitable ingredients can be appropriately selected and applied from formulation additives commonly used in the art. The antibody-drug conjugate used in the present disclosure can be administered, for example, as a pharmaceutical product containing a buffer such as a histidine buffer, a vehicle such as sucrose and trehalose, and a surfactant such as polysorbate 80 and 20. The antibody-drug conjugate used in the pharmaceutical products of the present disclosure can preferably be used as an injection, more preferably as an aqueous injection or a freeze-dried injection, and even more preferably as a freeze-dried injection. In the case where the pharmaceutical product containing the antibody-drug conjugate used in the present disclosure is an aqueous injection, the aqueous injection can preferably be diluted with a suitable diluent and then administered as an intravenous infusion. Examples of diluents can include dextrose solutions, physiological saline, etc., preferably dextrose solutions can be exemplified, and more preferably 5% dextrose solution can be exemplified. In the case where the pharmaceutical product of the present disclosure is a freeze-dried injection, the required amount of the freeze-dried injection is pre-dissolved in water for injection, preferably diluted with a suitable diluent, and then administered as an intravenous infusion. Examples of diluents can include dextrose solutions, physiological saline, etc., preferably dextrose solutions can be exemplified, and more preferably 5% dextrose solution can be exemplified.

[0172] Examples of administration routes applicable to administering the pharmaceutical products of the present disclosure include intravenous, intradermal, subcutaneous, intramuscular, and intraperitoneal routes, with the intravenous route being preferred.

[0173] The size of the dose required for the therapeutic treatment of a particular disease state will necessarily vary depending on the subject being treated, the administration route, and the severity of the disease to be treated. Further information regarding administration routes and dosage regimens can be found in Chapter 25.3, Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.

[0174] The anti-TROP2 antibody-drug conjugate used in the present disclosure can be administered to humans once at intervals of 1 to 180 days, and can preferably be administered once a week, once every two weeks, once every three weeks, once every four weeks, and even more preferably once every three weeks. Additionally, the antibody-drug conjugate used in the present disclosure can be administered at a dose of about 0.001 - 100 mg / kg, and can preferably be administered at a dose of 0.8 - 12.4 mg / kg. For example, the anti-TROP2 antibody-drug conjugate can be administered once every three weeks at a dose of about 0.27 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 4.0 mg / kg, 6.0 mg / kg, or 8.0 mg / kg, and can preferably be administered once every three weeks at a dose of 4.0 or 6.0 mg / kg.

[0175] The DNMT inhibitor can be administered at an appropriate dose via any appropriate route of administration.

[0176] In some aspects, the DNMT inhibitor is administered to a subject at a dose of about 0.1 - 10000 mg / m 2 body surface area per administration, and can preferably be administered at a dose of 15 - 75 mg / m 2 per administration, and even more preferably at a dose of 15 or 20 mg / m 2 per administration.

[0177] In some aspects, a dose of the DNMT inhibitor is administered to a subject daily for 2 - 7 days at intervals of 3 - 6 weeks, and can preferably be administered daily for 3 or 5 days at intervals of 6 or 4 weeks, and even more preferably at a dose of 20 mg / m 2 per day for 5 days at intervals of 4 weeks.

[0178] In some aspects, the DNMT inhibitor is administered in combination with one or more other chemotherapeutic agents. In some aspects, the other chemotherapeutic agent is cedazuridine. In some aspects, the chemotherapeutic agent is pemetrexed.

[0179] In some aspects, the DNMT inhibitors disclosed herein can be formulated with a pharmaceutically acceptable carrier, excipient, or stabilizer into a pharmaceutical composition. In certain aspects, the pharmaceutical composition is suitable for administration to a human or non-human animal via any one or more routes of administration known in the art. The term "pharmaceutically acceptable carrier" means one or more non-toxic materials that do not interfere with the biological activity effectiveness of the active ingredient. Such formulations routinely may contain salts, buffers, preservatives, compatible carriers, and optionally other therapeutic agents. Such pharmaceutically acceptable formulations may also contain compatible solid or liquid fillers, diluents, or encapsulating substances suitable for administration to humans. Other contemplated carriers, excipients, and / or additives that can be used in the formulations described herein include, for example, flavoring agents, antimicrobial agents, sweetening agents, antioxidants, antistatic agents, lipids, protein excipients (such as serum albumin, gelatin, casein), counterions forming salts (such as sodium), etc. Such and additional known pharmaceutical carriers, excipients, and / or additives suitable for the formulations described herein are known in the art, for example, as listed in "Remington: The Science & Practice of Pharmacy", 21st ed., Lippincott Williams & Wilkins, (2005), and in "Physician's Desk Reference", 60th ed., Medical Economics, Montvale, N.J. (2005). A pharmaceutically acceptable carrier can be selected that is suitable for the desired or required mode of administration, solubility, and / or stability.

[0180] In some aspects, the therapeutic composition can be formulated for a particular route of administration, such as oral, nasal, pulmonary, topical (including buccal and sublingual), rectal, vaginal, and / or parenteral administration. As used herein, the terms "parenteral administration" and "being administered parenterally" refer to modes of administration other than enteral and topical administration, usually via injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion. Formulations of the present disclosure suitable for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. Antibodies and other active substances can be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers, or propellants that may be required (see, for example, U.S. Patent Nos. 7,378,110; 7,258,873; and 7,135,180; U.S. Patent Application Publication Nos. 2004 / 0042972 and 2004 / 0042971).

[0181] The pharmaceutical composition can be in unit dosage form and can be prepared by any method known in the art of pharmacy. The actual dosage level of the active ingredient in the pharmaceutical composition of the present disclosure can be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient (e.g., "therapeutically effective amount"). The selected dosage level will depend on a variety of pharmacokinetic factors, including the activity of the particular composition employed, the route of administration, the time of administration, the rate of excretion of the particular compound used, the duration of the treatment, other drugs, the compounds and / or materials used in combination with the particular composition employed, the age, sex, weight, condition, general health, and prior medical history of the patient to be treated, and like factors well known in the medical arts. These dosages can be administered daily, weekly, bi-weekly, monthly, or less frequently, e.g., once every six months, depending on the dosage, the method of administration, the condition or symptoms to be treated, and the characteristics of the individual subject. The dosage can also be administered via continuous infusion (e.g., by pump). The dosage administered can also depend on the route of administration. For example, subcutaneous administration may require a higher dosage than intravenous administration. As described above, any conventional dosing regimen can be adapted (e.g., administering 1 - 10 mg / kg by injection or infusion once daily or twice weekly) and is suitable for methods related to treating human cancer patients.

[0182] Examples

[0183] The present disclosure is specifically described with reference to the examples shown below. However, the present disclosure is not limited by them. Further, it should in no way be construed in a limiting manner.

[0184] Example 1A: Preparation of Anti-TROP2 Antibody-Drug Conjugate (1)

[0185] According to the production methods described in WO2015 / 098099, WO2017 / 002776, and WO2022 / 014698, and using an anti-TROP2 antibody (an antibody comprising a heavy chain and a light chain, the heavy chain consisting of the amino acid sequence shown in SEQ ID NO:12 [= amino acid residues 20 - 470 of SEQ ID NO:1], and the light chain consisting of the amino acid sequence shown in SEQ ID NO:13 [= amino acid residues 21 - 234 of SEQ ID NO:2]), an anti-TROP2 antibody-drug conjugate (DS-1062a: deruxtecan) was prepared, in which the drug-linker shown in the following formula was conjugated to the anti-TROP2 antibody via a thioether bond:

[0186]

[0187] Among them, A represents the conjugation position with the antibody. The DAR of the antibody-drug conjugate (1) is 4.0.

[0188] Example 1B: Preparation of anti-TROP2 antibody-drug conjugate (2)

[0189] According to the manufacturing method described in Example 12 of US 7,999,083 and using the hRS7 antibody (an antibody comprising a heavy chain and a light chain, the heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 14, and the light chain comprising the amino acid sequence shown in SEQ ID NO: 15), sacituzumab govitecan (IMMU-132) was prepared. The DAR of the antibody-drug conjugate (2) is 7.5.

[0190] Example 2: Cell growth inhibition study

[0191] Combination of antibody-drug conjugate DS-1062a and decitabine

[0192] Human colorectal cancer cell lines DLD-1 and HCT-15 obtained from the American Type Culture Collection (ATCC) were incubated for 3 days at 37 °C, 5% CO2 in RPMI 1640 medium supplemented with 10% (v / v) heat-inactivated FBS, 1% (v / v) penicillin-streptomycin solution, and 1 mM sodium pyruvate (medium) with or without 1, 0.3, and 0.1 μM of decitabine (DAC) (Tokyo Chemical Industry Co., Ltd). The cells were harvested and re-seeded at 1000 cells / 80 μL / well in a 96-well black clear bottom plate and incubated overnight at 37 °C, 5% CO2. Then, 20 μL of a DS-1062a solution diluted with medium at concentrations of 500 nM - 0.5 nM was added to the wells, and the DLD-1 cells were incubated for 6 days and the HCT-15 cells were incubated for 8 days. The final concentration of DS-1062a was 100, 10, 1, and 0.1 nM. After incubation, the cell ATP level was measured using the CellTiter-Glo luminescent cell viability assay (Promega) and a microplate reader.

[0193] Cell viability was evaluated and defined as the percentage relative to the value of the cells not treated with DS-1062a in each group treated with or without DAC (N = 1 (in triplicate), mean ± SD).

[0194] The individual cell viability (%) was calculated by the following equation:

[0195] Cell viability (%) = 100 × T / C

[0196] T: The individual luminescence intensity of the wells treated with DS-1062a at each concentration

[0197] C: Average luminescence intensity of the holes treated with DS-1062a

[0198] Results:

[0199] As Figure 12A and 12B shown in, in both the DLD-1 and HCT-15 cell lines, the cell growth inhibitory activity of DS-1062a was enhanced in the DAC-treated cells compared to the untreated cells.

[0200] Example 3: Antitumor assay

[0201] Combination of the antibody-drug conjugate DS-1062a and decitabine

[0202] Female BALB / c-nu mice (The Jackson Laboratory Japan, Inc.) at 5-6 weeks of age were acclimated for 4 days before entering the study.

[0203] 3×10 6 DLD-1 cells suspended in saline were implanted subcutaneously into the flanks of the mice. The long and short axes of the tumors were measured twice a week with an electronic digital caliper, and the tumor volume was calculated by the following equation:

[0204] Tumor volume (mm 3 ) = 1 / 2 × long axis (mm) × [short axis (mm)] 2 .

[0205] When the tumor volume reached approximately 100-150 mm 3 , the tumor-bearing mice were randomly assigned to the treatment groups (day 0), as shown in Table 1:

[0206] Table 1

[0207]

[0208] QD: Administered once daily (quaquedie),

[0209] SC: Subcutaneous administration,

[0210] IV: Intravenous administration,

[0211] NT: Untreated

[0212] ABS: 10 mM acetate buffer [pH 5.5], 5% sorbitol

[0213] Decitabine (Tokyo Chemical Industry Co., Ltd) was dissolved and diluted with phosphate buffered saline (PBS), and administered subcutaneously to mice. The DS-1062a solution was diluted with ABS buffer (10 mM acetate buffer [pH 5.5], 5% sorbitol) and administered intravenously to the tail vein of mice. The dose of the compound for each mouse was calculated based on the individual body weight on the day of administration. From day 0 to day 4, decitabine was administered subcutaneously once a day at 0.5 mg / kg (total of 5 administrations). On day 7, DS-1062a was administered intravenously (single dose) to the tail vein at a liquid volume of 10 mL / kg at 10 mg / kg.

[0214] On the day of the final measurement (day 28), the Welch's t-test was used to evaluate the statistical significance between the "Ds-1062a" group and the "decitabine + Ds-1062a" group in terms of tumor volume.

[0215] Results:

[0216] As Figure 13 shown, compared with the untreated group, the anti-tumor activity of DS-1062a was significantly enhanced in the DAC pretreatment group (P < 0.05).

[0217] Example 4: Cell growth inhibition study

[0218] Combination of antibody-drug conjugate DS-1062a and azacitidine

[0219] The human colorectal cancer cell line DLD-1 was incubated for 3 days at 37 °C, 5% CO2 in RPMI 1640 medium supplemented with 10% (v / v) heat-inactivated FBS (medium) with or without 10, 3.3, and 1.1 μM azacitidine (AZA). The cells were harvested and re-seeded at 1000 cells / 90 μL / well in a 96-well black clear bottom plate and incubated overnight at 37 °C, 5% CO2. Then, 10 μL of the DS-1062a solution diluted with medium at a concentration of 1000 nM - 1 nM was added to the wells and incubated for 6 days. The final concentration of DS-1062a was 100, 10, 1, and 0.1 nM. After incubation, the cell ATP level was measured using the CellTiter-Glo Luminescent Cell Viability Assay (Promega Corporation) and a microplate reader.

[0220] Cell viability was evaluated and defined as the percentage relative to the value of the cells untreated with DS-1062a in each group treated with or without AZA (N = 1 (in triplicate), mean ± SD).

[0221] The viability (%) of single cells was calculated by the following equation:

[0222] Cell viability (%) = 100 × T / C

[0223] T: The single luminescence intensity of the wells treated with DS-1062a at each concentration

[0224] C: The average luminescence intensity of the wells treated with DS-1062a

[0225] Results:

[0226] As Figure 14 shown, the cell growth inhibitory activity of DS-1062a was enhanced in the cells treated with AZA compared with the cells untreated with AZA.

[0227] Example 5: Antitumor assay

[0228] Combination of antibody-drug conjugate IMMU-132 and decitabine

[0229] Five-week-old female BALB / c-nu mice (The Jackson Laboratory Japan, Inc.) were acclimated for 5 days before entering the study.

[0230] 3×10 6 DLD-1 cells suspended in saline were subcutaneously implanted into the flanks of the mice. The major and minor axes of the tumors were measured twice a week with an electronic digital caliper, and the tumor volume was calculated by the following equation:

[0231] Tumor volume (mm 3 ) = 1 / 2 × major axis (mm) × [minor axis (mm)] 2 .

[0232] When the tumor volume reached approximately 100 - 150 mm 3 , the tumor-bearing mice were randomly assigned to the treatment groups (day 0), as shown in Table 2.

[0233] On the final measurement day (day 28), Welch's t-test was used to evaluate the statistical significance between the IMMU-132 group and the decitabine + IMMU-132 group in terms of tumor volume.

[0234] Table 2

[0235]

[0236] QD: Administered once a day (quaquedie),

[0237] QW: Administered once a week (quaqueweek),

[0238] SC: Subcutaneous administration,

[0239] IV: Intravenous administration,

[0240] NT: Untreated

[0241] ABS: 10 mM acetate buffer [pH 5.5], 5% sorbitol

[0242] Dissolve decitabine (DAC) in phosphate-buffered saline (PBS) and administer it subcutaneously to mice. Dissolve IMMU-132 in ABS buffer (10 mM acetate buffer [pH 5.5], 5% sorbitol) and administer it intravenously to the tail vein of mice. Calculate the dosing volume of the compound for each mouse based on the most recent individual body weight measured within 2 days before dosing. From day 0 to day 4, DAC was administered subcutaneously once daily at 0.5 mg / kg (total of 5 doses). IMMU-132 was administered intravenously to the tail vein at 10 mg / kg on days 7 and 14 (weekly dosing × 2).

[0243] Results:

[0244] As Figure 15 shown, the anti-tumor activity of IMMU-132 pretreated with DAC was not significantly different from that of IMMU-132 not pretreated with DAC, while DS-1062a pretreated with DAC showed significantly enhanced anti-tumor activity as Figure 13 shown (Example 3).

[0245] The foregoing written specification is considered to be sufficient to enable those skilled in the art to practice these embodiments. The foregoing description and examples detail certain embodiments and describe the best mode contemplated by the inventors. However, it should be understood that, no matter how detailed the foregoing appears in the text, these embodiments can be practiced in many ways, and the claims include any equivalents thereof.

[0246] Sequence Listing Free Text

[0247] SEQ ID NO:1 - Amino acid sequence of the heavy chain of the anti-TROP2 antibody

[0248] SEQ ID NO:2 - Amino acid sequence of the light chain of the anti-TROP2 antibody

[0249] SEQ ID NO:3 - Amino acid sequence of heavy chain CDRH1 [= amino acid residues 50 - 54 of SEQ ID NO:1]

[0250] SEQ ID NO:4 - Amino acid sequence of heavy chain CDRH2 [= amino acid residues 69 - 85 of SEQ ID NO:1]

[0251] Amino acid sequence of SEQ ID NO:5 - heavy chain CDRH3 [= amino acid residues 118 - 129 of SEQ ID NO:1]

[0252] Amino acid sequence of SEQ ID NO:6 - light chain CDRL1 [= amino acid residues 44 - 54 of SEQ ID NO:2]

[0253] Amino acid sequence of SEQ ID NO:7 - light chain CDRL2 [= amino acid residues 70 - 76 of SEQ ID NO:2]

[0254] Amino acid sequence of SEQ ID NO:8 - light chain CDRL3 [= amino acid residues 109 - 117 of SEQ ID NO:2]

[0255] Amino acid sequence of SEQ ID NO:9 - heavy chain variable region [= amino acid residues 20 - 140 of SEQ ID NO:1]

[0256] Amino acid sequence of SEQ ID NO:10 - light chain variable region [= amino acid residues 21 - 129 of SEQ ID NO:2]

[0257] Amino acid sequence of SEQ ID NO:11 - heavy chain [= amino acid residues 20 - 469 of SEQ ID NO:1]

[0258] Amino acid sequence of SEQ ID NO:12 - heavy chain [= amino acid residues 20 - 470 of SEQ ID NO:1]

[0259] Amino acid sequence of SEQ ID NO:13 - light chain [= amino acid residues 21 - 234 of SEQ ID NO:2]

[0260] Amino acid sequence of the heavy chain of SEQ ID NO:14 - hRS7 antibody

[0261] Amino acid sequence of the light chain of SEQ ID NO:15 - hRS7 antibody

Claims

1. A pharmaceutical product comprising an antibody-drug conjugate and a DNMT inhibitor for combination administration, wherein, The antibody-drug conjugate is an antibody-drug conjugate in which the drug-linker represented by the following formula is conjugated to an anti-TROP2 antibody via a thioether bond: wherein A represents the position of attachment to the antibody.

2. The pharmaceutical product according to claim 1, wherein the anti-TROP2 antibody is an antibody comprising a heavy chain and a light chain, the heavy chain comprising CDRH1 consisting of the amino acid sequence shown in SEQ ID NO:3, CDRH2 consisting of the amino acid sequence shown in SEQ ID NO:4, and CDRH3 consisting of the amino acid sequence shown in SEQ ID NO:5, and the light chain comprising CDRL1 consisting of the amino acid sequence shown in SEQ ID NO:6, CDRL2 consisting of the amino acid sequence shown in SEQ ID NO:7, and CDRL3 consisting of the amino acid sequence shown in SEQ ID NO:

8.

3. The pharmaceutical product according to claim 2, wherein the anti-TROP2 antibody is an antibody comprising a heavy chain and a light chain, the heavy chain comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO:9, and the light chain comprising a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO:

10.

4. The pharmaceutical product according to claim 2 or 3, wherein the anti-TROP2 antibody is an antibody comprising a heavy chain and a light chain, the heavy chain consisting of the amino acid sequence shown in SEQ ID NO:12, and the light chain consisting of the amino acid sequence shown in SEQ ID NO:

13.

5. The pharmaceutical product according to claim 4, wherein the antibody lacks a lysine residue at the carboxyl terminus of the heavy chain.

6. The pharmaceutical product according to any one of claims 1-5, wherein the average number of units of the drug-linker conjugated to each antibody molecule in the antibody-drug conjugate is in the range of 3.5-4.

5.

7. The pharmaceutical product according to any one of claims 1-5, wherein the antibody-drug conjugate is datopotamab deruxtecan (DS-1062a).

8. The pharmaceutical product according to any one of claims 1-7, wherein the DNMT inhibitor is decitabine or azacitidine, or a pharmaceutically acceptable salt thereof.

9. The pharmaceutical product according to claim 8, wherein the DNMT inhibitor is decitabine or a pharmaceutically acceptable salt thereof.

10. The pharmaceutical product according to any one of claims 1-9, wherein the product is a composition, and the composition comprises an antibody-drug conjugate and a DNMT inhibitor for co-administration.

11. The pharmaceutical product according to any one of claims 1-9, wherein the product is a combination preparation, and the combination preparation comprises an antibody-drug conjugate and a DNMT inhibitor for sequential or separate co-administration.

12. The pharmaceutical product according to any one of claims 1-11, wherein the DNMT inhibitor is co-administered with a cytidine deaminase inhibitor.

13. The pharmaceutical product according to claim 12, wherein the cytidine deaminase inhibitor is cedazuridine or a pharmaceutically acceptable salt thereof.

14. The pharmaceutical product according to any one of claims 1-13, wherein the product is for treating cancer.

15. The pharmaceutical product according to claim 14, wherein the cancer is at least one selected from the following: breast cancer, lung cancer, colorectal cancer, gastric cancer, esophageal cancer, head and neck cancer, adenocarcinoma of the esophagogastric junction, biliary tract cancer, Paget's disease, pancreatic cancer, ovarian cancer, uterine carcinosarcoma, urothelial carcinoma, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumor, digestive tract stromal tumor, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, corpus cancer of uterus, renal cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, glioblastoma multiforme, osteosarcoma, sarcoma and melanoma, cervical cancer, uterine cancer, testicular cancer and renal cell carcinoma.

16. The pharmaceutical product according to claim 15, wherein the cancer is colorectal cancer.

17. The pharmaceutical product according to claim 15, wherein the cancer is lung cancer.

18. The pharmaceutical product according to claim 17, wherein the lung cancer is non-small cell lung cancer.

19. The pharmaceutical product according to claim 15, wherein the cancer is breast cancer.

20. The pharmaceutical product according to any one of claims 14-19, wherein the cancer cells of the cancer are SLFN11-deficient.

21. The pharmaceutical product according to claim 20, wherein the SLFN11 expression in the cancer cells of the patient is lower than that in the non-cancer cells expressing SLFN11 of the patient.

22. An antibody-drug conjugate for use in combination with a DNMT inhibitor in the treatment of cancer, wherein the antibody-drug conjugate and the DNMT inhibitor are as defined in any one of claims 1-9.

23. The antibody-drug conjugate for use according to claim 21, wherein the cancer is as defined in any one of claims 15-21.

24. The antibody-drug conjugate for use according to claim 22 or 23, wherein the use comprises sequential administration of the antibody-drug conjugate and the DNMT inhibitor.

25. The antibody-drug conjugate for use according to claim 22 or 23, wherein the use comprises separate and simultaneous administration of the antibody-drug conjugate and the DNMT inhibitor.

26. A method for treating cancer, which comprises co-administering an antibody-drug conjugate and a DNMT inhibitor as defined in any one of claims 1-9 to a subject in need thereof.

27. The method according to claim 26, wherein the cancer is as defined in any one of claims 15-21.

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