Combination of an Antibody-Drug Conjugate and a Bispecific Checkpoint Inhibitor

Abstract

A pharmaceutical product is provided for the administration of an antibody-drug conjugate in combination with a bispecific checkpoint inhibitor. The antibody-drug conjugate is an antibody-drug conjugate in which a drug-linker represented by the following formula (wherein A represents the position of connection to the antibody) is conjugated to an antibody, preferably an anti-TROP2 antibody or an anti-HER2 antibody, via a thioether bond. Also provided are therapeutic uses and methods in which the antibody-drug conjugate and the bispecific checkpoint inhibitor are administered in combination to a subject (Formula I). JPEG2025524992000015.jpg43128

Description

Technical Field

[0001] The present disclosure relates to a pharmaceutical product for the administration of a specific antibody-drug conjugate having an antitumor drug conjugated, via a linker structure, preferably to an anti-TROP2 or anti-HER2 antibody, in combination with a bispecific checkpoint inhibitor, and to the therapeutic use and method in which a specific antibody-drug conjugate and a bispecific checkpoint inhibitor are administered in combination to a subject.

Background Art

[0002] Immune checkpoint inhibitors are agents that inhibit the immunosuppressive system and activate antitumor immunity (Menon S. et al., Cancers (2016) 8, 106; Pardoll DM., Nat Rev Cancer (2012) 12, 252-264; Wolchok JD., Cell (2015) 162, 937). Well-validated targets for immune-mediated therapy in oncology include PD-1 (programmed cell death protein-1) and CTLA-4 (cytotoxic T lymphocyte-associated protein 4). TIGIT (T cell immunoreceptor with Ig and ITIM domains), an immunoreceptor present on some T cells and natural killer (NK) cells, is another target. Examples of known immune checkpoint inhibitors include the anti-PD-1 antibodies nivolumab (International Publication No. WO2006 / 121168) and pembrolizumab (International Publication No. WO2008 / 156712); the anti-PD-L1 antibodies atezolizumab (International Publication No. WO2010 / 077634), durvalumab (International Publication No. WO2011 / 066389), and avelumab (International Publication No. WO2013 / 079174); and the anti-CTLA-4 antibodies ipilimumab (International Publication No. WO2001 / 014424) and tremelimumab (International Publication No. WO2000 / 037504).

[0003] Bispecific binding proteins that specifically bind to two immune checkpoint targets are under development. Examples include the PD-1 / CTLA-4 bispecific antibody AK104 (cadonilimab) and MEDI5752 (U.S. Patent No. 10,457,732), which contain a first domain that specifically binds to PD-1 and a second domain that specifically binds to CTLA-4.

[0004] Antibody-drug conjugates (ADCs) composed of a cytotoxic drug conjugated to an antibody can selectively deliver the drug to and 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 HA., et al., J. Clin. Oncol. (2011) 29(4):398-405).

[0005] One such antibody-drug conjugate is trastuzumab deruxtecan, which is composed of a HER2-targeted antibody and a derivative of exatecan (Ogitani Y.et al., Clinical Cancer Research (2016) 22(20), 5097-5108; Ogitani Y.et al., Cancer Science (2016) 107, 1039-1046). Trastuzumab deruxtecan (Enhertu®, DS-8201) has shown significant clinical efficacy in HER2-expressing solid tumors, including breast cancer, gastric cancer, colorectal cancer, and non-small cell lung cancer. Importantly, DS-8201 has shown promising activity in HER2-low tumors in the above indications.

[0006] Another such antibody-drug conjugate is datopotamab deruxtecan (DS-1062), which is composed of a TROP2-targeting antibody and a derivative of exatecan. In particular, WO 2015 / 098099 and WO 2020 / 240467 provide a detailed description of exemplary TROP2-targeted antibody-drug conjugates including datopotamab deruxtecan (DS-1062). Datopotamab deruxtecan has shown clinical efficacy in multiple tumor types including lung cancer and breast cancer.

[0007] References disclosing the combined administration of antibody-drug conjugates and immune checkpoint inhibitors include Muller P. et al., Science Translational Medicine (2015) 7(315), 315ra188) (trastuzumab emtansine (T-DM1) in combination with both anti-CTLA-4 and anti-PD-1 antibodies); and WO 2018 / 110515 (trastuzumab deruxtecan (DS-8201) in combination with anti-PD-1, anti-PD-L1, anti-CD4 and anti-CD8 antibodies).

[0008] However, there is a need to identify further combination partners for antibody-drug conjugates, including those for anti-HER2 antibody-drug conjugates such as DS-8201 and anti-TROP2 antibody-drug conjugates such as DS-1062, to enhance their therapeutic capabilities.

[0009] Despite the therapeutic potential of antibody-drug conjugates such as DS-8201 and DS-1062 as monotherapies or in combination with checkpoint inhibitors, and the therapeutic potential of bispecific checkpoint inhibitors, 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. SUMMARY OF THE INVENTION

[0010] Antibody-drug conjugates (e.g., anti-TROP2 or anti-HER2 antibody-drug conjugates) used in the present disclosure, which contain a derivative of the topoisomerase I inhibitor exatecan as a component, have been confirmed to exhibit excellent antitumor effects in the treatment of certain cancers such as breast cancer when administered alone or in combination with checkpoint inhibitors. Furthermore, bispecific checkpoint inhibitors have been confirmed to exhibit antitumor effects in the treatment of certain cancers. However, there is a desire to provide pharmaceuticals and treatments that can achieve excellent antitumor effects in the treatment of cancer, such as enhanced efficacy, increased durability of treatment response, and / or decreased dose-dependent toxicity.

[0011] The present disclosure provides a pharmaceutical product that can exhibit excellent antitumor effects in the treatment of cancer by administering an antibody-drug conjugate, preferably an anti-TROP2 or anti-HER2 antibody-drug conjugate, in combination with a bispecific checkpoint inhibitor, preferably an anti-PD-1 / CTLA-4 or anti-PD-1 / TIGIT bispecific binding protein. The present disclosure also provides a therapeutic use and method in which an antibody-drug conjugate and a bispecific checkpoint inhibitor are administered in combination to a subject.

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

[93] : [1] A pharmaceutical product containing an antibody-drug conjugate and a bispecific checkpoint inhibitor for combined administration, wherein the antibody-drug conjugate has a drug-linker represented by the following formula:

[0013]

Chemical formula

[10] The pharmaceutical product according to [9], wherein the anti-HER2 antibody comprises a heavy chain comprising CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 16, CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 17, and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 18, and a light chain comprising CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 19, CDRL2 consisting of the amino acid sequence consisting of amino acid residues 1 to 3 of SEQ ID NO: 20, and CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 21;

[11] The pharmaceutical product according to

[10] , wherein the anti-HER2 antibody is an antibody comprising a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22 and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 23;

[12] The pharmaceutical product according to

[11] , wherein the anti-HER2 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 14 and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 15;

[13] The pharmaceutical product according to

[11] , wherein the anti-HER2 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 24 and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 15;

[14] The pharmaceutical product according to any one of [9] to

[13] , wherein the average number of conjugated drug-linkers per anti-HER2 antibody molecule in the antibody-drug conjugate ranges from 7 to 8;

[15] The pharmaceutical product according to

[14] , wherein the anti-HER2 antibody-drug conjugate is trastuzumab deruxtecan (DS-8201);

[16] The pharmaceutical product according to any one of [1] to

[15] , wherein the bispecific checkpoint inhibitor is a bispecific binding protein comprising a first binding domain that specifically binds to PD-1 and a second binding domain that specifically binds to CTLA-4 or TIGIT;

[17] The bispecific binding protein is a) A first binding domain that specifically binds to PD-1, comprising a heavy chain variable domain containing CDRH1 having the amino acid sequence of SEQ ID NO: 25, CDRH2 having the amino acid sequence of SEQ ID NO: 26, and CDRH3 having the amino acid sequence of SEQ ID NO: 27, and a light chain variable domain containing CDRL1 having the amino acid sequence of SEQ ID NO: 28, CDRL2 having the amino acid sequence of SEQ ID NO: 29, and CDRL3 having the amino acid sequence of SEQ ID NO: 30, and the first binding domain; b) A second binding domain that specifically binds to TIGIT, comprising a heavy chain variable domain containing CDRH1 having the amino acid sequence of SEQ ID NO: 35, CDRH2 having the amino acid sequence of SEQ ID NO: 36, and CDRH3 having the amino acid sequence of SEQ ID NO: 37, and a light chain variable domain containing CDRL1 having the amino acid sequence of SEQ ID NO: 38, CDRL2 having the amino acid sequence of SEQ ID NO: 39, and CDRL3 having the amino acid sequence of SEQ ID NO: 40, and the second binding domain, and the pharmaceutical product according to

[16] ;

[18] The pharmaceutical product according to

[17] , wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 31 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 33;

[19] The pharmaceutical product according to

[17] , wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 31 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 33;

[20] The pharmaceutical product according to any one of

[17] to

[19] , wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 32 and a light chain having the amino acid sequence of SEQ ID NO: 34;

[21] The pharmaceutical product according to any one of

[17] to

[19] , wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 32 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 34;

[22] A pharmaceutical product according to any one of

[17] to

[21] , wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 43;

[23] A pharmaceutical product according to any one of

[17] to

[21] , wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 43;

[24] A pharmaceutical product according to any one of

[17] to

[23] , wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino acid sequence of SEQ ID NO: 42 and a light chain having the amino acid sequence of SEQ ID NO: 44;

[25] A pharmaceutical product according to any one of

[17] to

[23] , wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 42 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 44;

[26] The bispecific binding protein is a) a first binding domain that specifically binds to PD-1, the first binding domain comprising a heavy chain variable domain comprising CDRH1 having the amino acid sequence of SEQ ID NO: 52, CDRH2 having the amino acid sequence of SEQ ID NO: 53, and CDRH3 having the amino acid sequence of SEQ ID NO: 54, and a light chain variable domain comprising CDRL1 having the amino acid sequence of SEQ ID NO: 49, CDRL2 having the amino acid sequence of SEQ ID NO: 50, and CDRL3 having the amino acid sequence of SEQ ID NO: 51; b) A second binding domain that specifically binds to CTLA-4, comprising a heavy chain variable domain comprising CDRH1 having the amino acid sequence of SEQ ID NO: 58, CDRH2 having the amino acid sequence of SEQ ID NO: 59, and CDRH3 having the amino acid sequence of SEQ ID NO: 60, and a light chain variable domain comprising CDRL1 having the amino acid sequence of SEQ ID NO: 55, CDRL2 having the amino acid sequence of SEQ ID NO: 56, and CDRL3 having the amino acid sequence of SEQ ID NO: 57; a second binding domain; the pharmaceutical product according to

[16] ;

[27] The pharmaceutical product according to

[26] , wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 46 and a light chain having the amino acid sequence of SEQ ID NO: 45;

[28] The pharmaceutical product according to

[26] , wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 46 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 45;

[29] The pharmaceutical product according to any one of

[26] to

[28] , wherein the second binding domain that specifically binds to CTLA-4 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 48 and a light chain having the amino acid sequence of SEQ ID NO: 47;

[30] The pharmaceutical product according to any one of

[26] to

[28] , wherein the second binding domain that specifically binds to CTLA-4 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 48 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 47;

[31] The pharmaceutical product according to

[20] or

[27] , wherein the light chain constant region is a kappa chain;

[32] The pharmaceutical product according to

[24] or

[29] , wherein the light chain constant region is a lambda chain;

[33] The pharmaceutical product according to any one of

[16] to

[32] , wherein the binding protein is an antibody;

[34] The pharmaceutical product according to

[33] , wherein the antibody is an IgG antibody;

[35] The pharmaceutical product according to

[34] , wherein the antibody is an IgG1 antibody;

[36] The pharmaceutical product according to

[34] or

[35] , wherein the antibody is a human antibody or a humanized antibody;

[37] The pharmaceutical product according to any one of

[33] to

[36] , wherein the bispecific antibody is monovalent;

[38] The pharmaceutical product according to any one of

[16] to

[37] , wherein the bispecific binding protein is DuetMab;

[39] The pharmaceutical product according to any one of

[16] to

[38] , wherein the bispecific binding protein contains a mutated Fc region;

[40] When the mutant Fc region is numbered according to the EU index described in Kabat, 221K, 221Y, 225E, 225K, 225W, 228P, 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235E, 235F, 236E, 237L, 237M, 237P, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 240I, 240A, 240T, 240M, 241W, 241L, 241Y, 241E, 241R, 243W, 243L, 243Y, 243R, 243Q, 244H, 245A, 247L, 247V, 247G, 250E, 250Q, 251F, 252L, 252Y, 254S, 254T, 255L, 256E, 256F, 256M, 257C, 257M, 257N, 262I, 262A, 262T, 262E, 263I, 263A, 263T, 263M, 264L, 264I, 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265A, 265G, 265N, 265Q, 265Y, 265F, 265V, 265I, 265L, 265H, 265T, 266I, 266A, 266T, 266M, 267Q, 267L, 268E, 269H, 269Y, 269F, 269R, 270E, 280A, 284M, 292P, 292L, 296E, 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H, 296G, 297S, 297D, 297E, 298A, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 305I, 308F, 313F, 316D, 318A, 318S, 320A, 320S, 322A, 322S, 325Q, 325L, 3251, 325D, 325E, 325A, 325T, 325V, 325H, 326A, 326D, 326E, 326G, 326M, 326V, 327G, 327W, 327N, 327L, 328S, 328M, 328D, 328E, 328N, 328Q, 328F, 3281, 328V, 328T, 328H, 328A, 329F, 329H, 329Q, 330K,The pharmaceutical product according to

[39] , comprising at least one substitution selected from 330G, 330T, 330C, 330L, 330Y, 330V, 3301, 330F, 330R, 330H, 331G, 331A, 331L, 331M, 331F, 331W, 331K, 331Q, 331E, 331S, 331V, 3311, 331C, 331Y, 331H, 331R, 331N, 331D, 331T, 332D, 332S, 332W, 332F, 332E, 332N, 332Q, 332T, 332H, 332Y, 332A, 333A, 333D, 333G, 333Q, 333S, 333V, 334A, 334E, 334H, 334L, 334M, 334Q, 334V, 334Y, 339T, 370E, 370N, 378D, 392T, 396L, 416G, 419H, 421K, 428L, 428F, 433K, 433L, 434A, 434W, 434Y, 436H, 440Y and 443W; The pharmaceutical product according to

[39] , wherein the mutant Fc region comprises one or more amino acid substitutions at positions selected from 428 and 434 numbered according to the EU index described in Kabat; The pharmaceutical product according to any one of

[39] to

[41] , wherein the mutant Fc region comprises one or more amino acid substitutions selected from 428L, 428F, 434A, 434W, and 434Y; The pharmaceutical product according to any one of

[39] to

[42] , wherein the mutant Fc region comprises the L234F / L235E / P331S triple mutation (TM); The pharmaceutical product according to any one of

[16] to

[43] , wherein the bispecific binding protein comprises a non-glycosylated Fc region; The pharmaceutical product according to any one of

[16] to

[43] , wherein the bispecific binding protein comprises a deglycosylated Fc region; The pharmaceutical product according to any one of

[16] to

[43] , wherein the bispecific binding protein comprises an Fc region having reduced fucosylation or no fucosylation;

[47] The pharmaceutical product according to any one of [1] to

[46] , wherein the product is a combination preparation comprising an antibody-drug conjugate and a bispecific checkpoint inhibitor for separate simultaneous administration;

[48] The pharmaceutical product according to any one of [1] to

[46] , wherein the product is a combination preparation comprising an antibody-drug conjugate and a bispecific checkpoint inhibitor for sequential or separate simultaneous administration;

[49] The pharmaceutical product according to any one of [1] to

[48] , wherein the product is for treating cancer;

[50] The pharmaceutical product according to

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

[51] The pharmaceutical product according to

[50] , wherein the cancer is breast cancer;

[52] The pharmaceutical product according to

[51] , wherein the breast cancer is HER2-positive breast cancer;

[53] The pharmaceutical product according to

[51] , wherein the breast cancer is HER2-low breast cancer;

[54] The pharmaceutical product according to

[51] , wherein the breast cancer is triple-negative breast cancer;

[55] The pharmaceutical product according to

[51] , wherein the breast cancer is hormone receptor (HR)-positive, HER2-negative breast cancer;

[56] The pharmaceutical product according to

[50] , wherein the cancer is lung cancer;

[57] The pharmaceutical product according to

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

[58] The pharmaceutical product according to

[57] , wherein the non-small cell lung cancer is non-small cell lung cancer having actionable genomic alterations;

[59] The pharmaceutical product according to

[57] , wherein the non-small cell lung cancer is non-small cell lung cancer having no actionable genomic alterations; The pharmaceutical product according to

[50] , wherein the cancer is colorectal cancer; The pharmaceutical product according to

[50] , wherein the cancer is gastric cancer; The pharmaceutical product according to

[50] , wherein the cancer is pancreatic cancer; The pharmaceutical product according to

[50] , wherein the cancer is ovarian cancer; The pharmaceutical product according to

[50] , wherein the cancer is prostate cancer; The pharmaceutical product according to

[50] , wherein the cancer is renal cancer; The pharmaceutical product according to

[50] , wherein the cancer is bladder cancer; The pharmaceutical product according to

[50] , wherein the cancer is endometrial cancer; The pharmaceutical product according to

[50] , wherein the cancer is bile duct cancer; The pharmaceutical product according to any one of [1] to

[48] for use in the treatment of cancer; The pharmaceutical product for use according to

[69] , wherein the cancer is as described in any one of

[50] to

[68] ; The pharmaceutical product according to any one of [1] to

[48] , further comprising carboplatin for administration in combination with an antibody-drug conjugate and a bispecific checkpoint inhibitor; The pharmaceutical product according to any one of [1] to

[48] , further comprising a fluoropyrimidine for administration in combination with an antibody-drug conjugate and a bispecific checkpoint inhibitor; Use of an antibody-drug conjugate in the manufacture of a medicament for use in combination with a bispecific checkpoint inhibitor, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as described in any one of [1] to

[46] for treating cancer; The use according to

[73] , wherein the medicament is for use in combination with a bispecific checkpoint inhibitor by continuous administration; The use according to

[73] , wherein the medicament is for use in combination with a bispecific checkpoint inhibitor by separate simultaneous administration;

[76] Use of a bispecific checkpoint inhibitor in the manufacture of a medicament for use in combination with an antibody-drug conjugate, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as described in any one of [1] to

[46] for treating cancer;

[77] The use according to

[76] , wherein the medicament is for use in combination with an antibody-drug conjugate by continuous administration;

[78] The use according to

[76] , wherein the medicament is for use in combination with an antibody-drug conjugate by separate simultaneous administration;

[79] The use according to any one of

[73] to

[78] , wherein the cancer is as described in any one of

[50] to

[68] ;

[80] An antibody-drug conjugate for use in combination with a bispecific checkpoint inhibitor in the treatment of cancer, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as described in any one of [1] to

[46] ;

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

[80] , wherein the cancer is as described in any one of

[50] to

[68] ;

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

[80] or

[81] , wherein the use comprises continuously administering the antibody-drug conjugate and the bispecific checkpoint inhibitor;

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

[80] or

[81] , wherein the use comprises administering the antibody-drug conjugate and the bispecific checkpoint inhibitor separately or simultaneously;

[84] An antibody-drug conjugate for use in the treatment of cancer in a subject, wherein the treatment comprises i) continuous or separate simultaneous administration to the subject of the antibody-drug conjugate and ii) a bispecific checkpoint inhibitor, and the antibody-drug conjugate and the bispecific checkpoint inhibitor are as described in any one of [1] to

[46] ;

[85] A bispecific checkpoint inhibitor for use in combination with an antibody-drug conjugate in the treatment of cancer, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as described in any one of [1] to

[46] ; a bispecific checkpoint inhibitor.

[86] The bispecific checkpoint inhibitor for use according to

[85] , wherein the cancer is as described in any one of

[50] to

[68] ;

[87] The bispecific checkpoint inhibitor for use according to

[85] or

[86] , wherein the use comprises the sequential administration of an antibody-drug conjugate and a bispecific checkpoint inhibitor;

[88] The bispecific checkpoint inhibitor for use according to

[85] or

[86] , wherein the use comprises the separate or simultaneous administration of an antibody-drug conjugate and a bispecific checkpoint inhibitor;

[89] A bispecific checkpoint inhibitor for use in the treatment of cancer in a subject, wherein the treatment comprises i) the sequential or separate simultaneous administration to the subject of a bispecific checkpoint inhibitor and ii) an antibody-drug conjugate, and the bispecific checkpoint inhibitor and the antibody-drug conjugate are as described in any one of [1] to

[46] ; a bispecific checkpoint inhibitor.

[90] A method for treating cancer, comprising administering in combination to a subject in need thereof an antibody-drug conjugate and a bispecific checkpoint inhibitor as described in any one of [1] to

[46] ;

[91] The method according to

[90] , wherein the cancer is as described in any one of

[50] to

[68] ;

[92] The method according to

[90] or

[91] , wherein the method comprises the sequential administration of an antibody-drug conjugate and a bispecific checkpoint inhibitor;

[93] The method according to

[90] or

[91] , wherein the method comprises the separate and simultaneous administration of an antibody-drug conjugate and a bispecific checkpoint inhibitor. [Effect of the Invention]

[0014] The present disclosure provides a pharmaceutical product for administering in combination a specific antibody-drug conjugate having an anti-tumor drug conjugated to an antibody (preferably an anti-TROP2 or anti-HER2 antibody) via a linker structure and a bispecific checkpoint inhibitor, and a therapeutic use and method for administering the specific antibody-drug conjugate and the bispecific checkpoint inhibitor in combination to a subject. Accordingly, the present disclosure provides a pharmaceutical and a treatment capable of obtaining an excellent anti-tumor effect in the treatment of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] [Anti-TROP2 antibody]:

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DETAILED DESCRIPTION OF THE INVENTION

[0016] To make the present disclosure more readily understandable, certain terms are first defined. Further definitions are set forth throughout the detailed description.

[0017] Unless otherwise defined, all scientific and technical 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 many common dictionaries of terms used in the present disclosure to those of ordinary skill in the art.

[0018] Unless the context requires otherwise, singular terms shall include the plural and plural terms shall include the singular.

[0019] Units, prefixes, and symbols are shown in the form approved by the International System of Units (SI). Numerical ranges include the numbers defining the range.

[0020] Whenever an aspect is described herein using the word "comprising", it is understood that other similar aspects described in terms of "consisting of" and / or "consisting essentially of" are also provided.

[0021] The terms "inhibit" and "inhibition" can refer to a decrease of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% of biological activity. Cell proliferation can be assayed using techniques recognized in the art that measure the rate of cell division, and / or the proportion of cells within a cell population undergoing cell division, and / or the rate of cell loss from a cell population due to terminal differentiation or cell death (e.g., thymidine incorporation).

[0022] The term "subject" refers to any animal (e.g., a mammal) including, but not limited to, humans, non-human primates, rodents, etc., that is the recipient of a particular treatment. Typically, the terms "subject" and "patient" are used interchangeably herein with respect to human subjects.

[0023] The term "pharmaceutical product" refers to a composition containing all active ingredients (for co-administration), or a combination of separate compositions, each containing at least one but not all of the active ingredients (combination preparation) (for sequential or simultaneous administration), in a form that enables the biological activity of the active ingredients, and which does not contain additional ingredients that are unacceptably toxic to the subject to whom the product is administered. Such a product may be sterile. "Co-administration" means that the active ingredients are administered simultaneously. "Sequential administration" means that the active ingredients are administered one after another, in any order, with a time interval between individual administrations. The time interval can be, for example, less than 24 hours, preferably less than 6 hours, more preferably less than 2 hours.

[0024] Terms such as "treat" or "treatment" or "for treating" or "alleviate" or "for alleviating" refer to both (1) therapeutic means that cure, slow down, relieve, and / or halt the progression of the symptoms of a diagnosed pathological condition or disorder, and (2) prophylactic or preventive means that prevent and / or delay the onset of a targeted pathological condition or disorder. Thus, those in need of treatment include those who already have a disorder; patients who tend to have a disorder; and patients in whom the disorder is to be prevented. In certain embodiments, when a patient exhibits, for example, a complete, partial, or transient remission of a particular type of cancer, the subject is successfully "treated" for the cancer according to the methods of the present disclosure.

[0025] The terms "cancer", "tumor", "cancerous", and "malignant" typically refer to or describe a physiological state in mammals characterized by unregulated cell growth. Examples of cancer include, but are not limited to, breast cancer, lung cancer, colorectal cancer, gastric cancer, esophageal cancer, head and neck cancer, esophagogastric junction adenocarcinoma, biliary tract cancer, Paget's disease, pancreatic cancer, ovarian cancer, uterine carcinosarcoma, urothelial cancer, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumor, gastrointestinal stromal tumor, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, corpus cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, multiple myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer, and renal cell carcinoma. Cancers include hematological malignancies such as acute myeloid leukemia, multiple myeloma, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, Burkitt lymphoma, follicular lymphoma, and solid tumors such as breast cancer, lung cancer, neuroblastoma, and colon cancer.

[0026] As used herein, the term "cytotoxic drug" is broadly defined to refer to a substance that inhibits or prevents the function of cells and / or causes the destruction of cells (cell death) and / or exerts an antineoplastic / antiproliferative effect. For example, cytotoxic drugs directly or indirectly prevent the development, maturation, or spread of neoplastic tumor cells. This term also includes agents that cause only a cytostatic effect, i.e., an effect that inhibits cell growth only. This term includes the chemotherapeutic agents identified below.

[0027] The term "chemotherapeutic agent" is a subset of the term "cytotoxic drug" and includes natural or synthetic chemical compounds.

[0028] According to the methods or uses of the present disclosure, the compounds of the present disclosure can be administered to a patient to promote a positive therapeutic response regarding cancer. The term "positive therapeutic response" regarding cancer treatment refers to the improvement of symptoms associated with the disease. For example, the improvement of the disease can be characterized as a complete response. The term "complete response" refers to the absence of clinically detectable disease, accompanied by the normalization of any previous test results. Alternatively, the improvement of the disease can be classified as a partial response. "Positive therapeutic response" includes the reduction or inhibition of cancer progression and / or duration, the reduction or improvement of cancer severity, and / or the improvement of one or more of its symptoms resulting from the administration of the compounds of the present disclosure. In certain embodiments, such terms refer to one, two, or more than two results after the administration of the compounds of the present disclosure: (1) Stabilization, reduction, or elimination of a cancer cell population; (2) Stabilization or reduction of cancer growth; (3) Impairment of cancer formation; (4) Eradication, removal, or control of primary, local, and / or metastatic cancer; (5) Reduction in mortality; (6) Increase in disease-free, recurrence-free, progression-free, and / or overall survival duration, persistence, or rate; (7) Increase in response rate, durability of response, or number of patients responding or in remission; (8) Reduction in hospitalization rate, (9) Decrease in hospitalization duration, (10) The cancer size is maintained, does not increase, or increases by less than 10%, preferably less than 5%, preferably less than 4%, preferably less than 2%, and (11) Increase in the number of patients in remission. (12) Decrease in the number of adjuvant therapies (e.g., chemotherapy or hormone therapy) that would otherwise be required to treat the cancer.

[0029] Clinical response, which is not limited to these, can include changes detectable by techniques such as ELISA, RIA, chromatography, etc., and can be evaluated using screening techniques, such as PET, magnetic resonance imaging (MRI) scans, X-ray imaging, computed tomography (CT) scans, flow cytometry or fluorescence-activated cell sorter (FACS) analysis, histology, gross pathology, and blood chemistry. In addition to these positive treatment responses, subjects undergoing treatment can experience the beneficial effect of improvement in symptoms associated with the disease.

[0030] As used herein, the term "antibody" refers to a protein capable of specifically binding by recognizing an antigen. Normal or conventional mammalian antibodies contain a tetramer, which typically consists of two identical pairs of polypeptide chains, and each pair consists of one "light" chain (typically having a molecular weight of about 25 kDa) and one "heavy" chain (typically 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 a variable domain sequence sufficient to confer specificity for the target antigen. The amino-terminal portion of each light and heavy chain typically contains a variable domain of about 100 - 110 or more amino acids typically involved in 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 typically defines a constant domain responsible for effector functions. Thus, in a naturally occurring antibody, a full-length heavy-chain immunoglobulin polypeptide contains a variable domain (V H ) and three constant domains (C H1 , C H2 , and C H3 ), as well as a hinge region between C H1 and C H2 . The VH domain is at the amino terminus of the polypeptide, the C H3 domain is at the carboxyl terminus, and a full-length light-chain immunoglobulin polypeptide contains a variable domain (V L ) and a constant domain (C L) including V L The domain is at the amino terminus of the polypeptide, and C L The domain is at the carboxyl terminus. However, one of ordinary skill in the art will understand that the positions of the domains in a naturally occurring antibody can be modified in certain antibody-like binding protein formats without loss of antigen-binding ability. The classes of human light chains are called kappa light chains and lambda light chains.

[0031] Within the full-length light and heavy chains, the variable and constant domains are typically linked 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 chain / heavy chain pair typically form the antigen-binding site. The variable domains of a naturally occurring antibody typically exhibit the same general structure of relatively conserved framework regions (FRs) linked by three hypervariable regions, also called complementarity-determining regions or CDRs. The CDRs from each pair of the two chains are typically aligned by the framework regions, which can allow for binding to a specific epitope. From the amino terminus to the carboxyl terminus, both the light chain variable domain and the heavy chain variable domain typically include the domains FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0032] The term "antibody fragment" refers to intact or full-length chains or parts of an antibody, generally 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, with respect to an antibody, can refer to antibody fragments such as F v , F ab , F (ab’)2 and the like.

[0033] References to the numbering of amino acid residues described herein are made according to the EU numbering system (and are also described in Kabat et al, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)).

[0034] A "monoclonal" antibody or antigen-binding fragment thereof refers to a homogeneous population of antibodies or antigen-binding fragments that are highly specific for binding to a single antigenic determinant or epitope. This is in contrast to polyclonal antibodies, which typically contain different antibodies against different antigenic determinants. The term "monoclonal" antibody or antigen-binding fragment thereof encompasses both intact monoclonal antibodies and full-length monoclonal antibodies, as well as antibody fragments (e.g., Fab, Fab’, F(ab’)2, Fv), single-chain (scFv) variants, fusion proteins containing antibody moieties, and any other modified immunoglobulin molecule containing an antigen recognition site. Further, a "monoclonal" antibody or antigen-binding fragment thereof refers to such antibodies and antigen-binding fragments produced in a manner including, but not limited to, hybridomas, phage selection, recombinant expression, and transgenic animals.

[0035] The term "human antibody", as used herein, encompasses antibodies having variable and constant regions that substantially correspond to human germline immunoglobulin sequences. In some embodiments, human antibodies are produced in non-human mammals including, but not limited to, rodents such as mice and rats, and lagomorphs such as rabbits. In other embodiments, human antibodies are produced in hybridoma cells. In yet other embodiments, human antibodies are produced recombinantly. In some embodiments, bispecific binding proteins are human or humanized antibodies.

[0036] As used herein, the terms "antigen" or "target antigen" refer to a molecule or a part of a molecule that can be recognized and bound by the binding proteins of the present disclosure. The target antigen can be used in an animal to produce an antibody that can bind to an epitope of the antigen. The target antigen can have one or more epitopes.

[0037] As used herein, the term "epitope" refers to a region or structural element of an antigen that is recognized and bound by the binding proteins of the present disclosure. More precisely, an epitope is a specific structure that is bound by the CDR of a binding protein. An epitope can include even a portion of a protein structural element, a carbohydrate, or a lipid structure present in a membrane. A binding protein is said to specifically bind to an antigen when it preferentially recognizes its antigen target in a complex mixture of proteins and / or macromolecules. The term "specifically binds" refers to a binding protein that specifically binds to a molecule or a fragment thereof (e.g., an antigen). A binding protein that specifically binds to a molecule or a fragment thereof can bind to other molecules with lower affinity, for example, as measured by an immunoassay, BIAcore, or other assays known in the art. In particular, an antibody or a fragment that specifically binds to at least one molecule or a fragment thereof can compete with molecules that bind non-specifically. The present disclosure specifically encompasses antibodies having multiple specificities (e.g., antibodies having specificities for two or more distinct antigens). For example, a bispecific antibody can bind to two adjacent epitopes on a single target antigen or can bind to two different antigens.

[0038] As used herein, the term "antigen-binding site" refers to a site created on the surface of a binding protein of the present disclosure to which an antigen or an epitope on an antigen binds. The antigen-binding site of a binding protein is typically described by referring to the loop structures created by the complementarity-determining regions (CDRs) of the binding protein.

[0039] [Description of Embodiments] Hereinafter, preferred embodiments for carrying out the present disclosure will be described. Note that the embodiments described below show examples of representative embodiments of the present disclosure and do not limit the scope of the present disclosure.

[0040] 1. Antibody-drug conjugate The antibody-drug conjugate used in the present disclosure is such that the drug-linker has the following formula:

[0041] [Chemical formula] It is an antibody-drug conjugate represented by In the formula, A represents the connection position to the antibody and is conjugated to the antibody, preferably an anti-TROP2 or anti-HER2 antibody, via a thioether bond.

[0042] In the present disclosure, the partial structure consisting of the linker and the drug in the antibody-drug conjugate is referred to as the "drug-linker". The drug-linker is linked to the thiol group (in other words, the sulfur atom of the cysteine residue) formed at the interchain disulfide bond sites (two sites between the heavy chains and two sites between the heavy chain and the light chain) in the antibody.

[0043] The drug-linker of the present disclosure contains exatecan (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 (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 and is a topoisomerase I inhibitor. Exatecan is a camptothecin derivative having an antitumor effect, represented by the following formula:

[0044] [Chemical formula]

[0045] The antibody-drug conjugates used in the present disclosure can also be represented by the following formula:

[0046]

Chemical formula

[0047] Here, the drug-linker is conjugated to an antibody (“antibody-”), preferably an anti-TROP2 or anti-HER2 antibody, via a thioether bond. The meaning of n is the so-called average number of conjugated drug molecules (DAR; drug-to-antibody ratio), which indicates the average number of units of the conjugated drug-linker per antibody molecule.

[0048] The antibody-drug conjugates used in the present disclosure move into cancer cells and are then cleaved at the linker moiety, releasing a compound represented by the following formula:

[0049]

Chemical formula

[0050] 2. Antibodies in Antibody-Drug Conjugates The antibodies in the antibody-drug conjugates used in the present disclosure are preferably anti-TROP2 or anti-HER2 antibodies and can be derived from any species, preferably human, rat, mouse, or rabbit. When the antibody is derived from a species other than human, it is preferably chimerized or humanized using known techniques. The antibody can be a polyclonal antibody or a monoclonal antibody, preferably a monoclonal antibody.

[0051] 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, for example, an antibody having the property of recognizing cancer cells, the property of binding to cancer cells, the property of internalizing into cancer cells, and / or the property of having cytotoxic activity against cancer cells.

[0052] The binding activity of the antibody to cancer cells can be confirmed using flow cytometry. The internalization of the antibody into cancer cells can be confirmed by (1) an assay for visualizing the antibody taken up into 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 taken up into 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 (Bio Techniques 28:162-165, January 2000) using an immunotoxin that binds to the therapeutic antibody and inhibits cell growth when the toxin is taken up into cells. As the immunotoxin, a recombinant complex protein of the diphtheria toxin catalytic domain and protein G can be used.

[0053] The antitumor activity of the antibody can be confirmed in vitro by measuring the inhibitory activity against cell proliferation. For example, a cancer cell line overexpressing the target protein of the antibody is cultured, and the antibody is added to the culture system at various concentrations to determine the inhibitory activity against focus formation, colony formation, and spheroid growth. The antitumor activity can be confirmed in vivo, for example, by administering the antibody to a mouse transplanted with a cancer cell line highly expressing the target protein and measuring the changes in the cancer cells.

[0054] In an antibody-drug conjugate, the compound conjugated therein preferably has an anti-tumor effect, but it is not essential for the antibody itself to have an anti-tumor effect. In order for the anti-tumor compound to specifically and selectively exhibit cytotoxic activity against cancer cells, it is important and preferable for the antibody to have the property of internalizing and migrating into cancer cells.

[0055] The antibody (preferably, anti-TROP2 or anti-HER2 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 include immunizing an animal with an antigenic polypeptide and collecting and purifying the antibodies produced in vivo. The origin of the antigen is not limited to humans, and animals may be immunized with antigens derived from non-human animals such as mice and rats. In this case, the cross-reactivity of the obtained antibody that binds to the heterologous antigen with human antigens can be tested to screen for antibodies applicable to human diseases.

[0056] Alternatively, antibody-producing cells that produce antibodies against an antigen can be fused with myeloma cells 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)) to establish hybridomas, from which monoclonal antibodies can be obtained in sequence.

[0057] The antigen can be obtained by genetically engineering a host cell to produce a gene encoding the antigen protein. Specifically, a vector for expressing the antigen gene is prepared and introduced into the host cell to express the gene. The antigen expressed in this way can be purified. Antibodies can also be obtained by a method of immunizing an animal with the above-mentioned genetically engineered antigen-expressing cells or cell lines expressing the antigen.

[0058] The antibodies (preferably anti-TROP2 or anti-HER2 antibodies) in the antibody-drug conjugates used in the present disclosure are preferably recombinant antibodies obtained by artificial modification for the purpose of reducing the heterologous antigenicity to humans, such as chimeric antibodies or humanized antibodies, or preferably antibodies having only the gene sequence of human-derived antibodies, that is, human antibodies. These antibodies can be produced using known methods.

[0059] Examples of chimeric antibodies include antibodies in which the variable region and the constant region of the antibody are derived from different species, for example, chimeric antibodies in which the antibody variable region derived from a mouse or a rat is linked to the antibody constant region derived from a human (Proc. Natl. Acad. Sci. USA, 81, 6851-6855, (1984)).

[0060] Examples of humanized antibodies include antibodies in which only the complementarity-determining regions (CDRs) of heterologous antibodies are incorporated into human-derived antibodies (Nature (1986) 321, pp. 522-525), antibodies in which a part of the amino acid residues of the framework of heterologous antibodies and the CDR sequences of heterologous antibodies are transplanted into human antibodies by the CDR transplantation method (International Publication No. 90 / 07861), antibodies humanized using the gene conversion mutagenesis method (U.S. Patent No. 5,821,337), and the like.

[0061] Examples of human antibodies include antibodies produced using human antibody-producing mice having human chromosomal fragments containing the genes for the heavy and light chains of human antibodies (see, for example, 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 method, antibodies obtained by phage display and selected from a human antibody library can be mentioned (see, for example, 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.).

[0062] The antibodies in the antibody-drug conjugates used in the present invention also include modified variants of the antibodies. The modified variants refer to variants obtained by chemically or biologically modifying the antibodies according to the present disclosure. Examples of chemically modified variants include variants in which a chemical moiety is bound to the amino acid backbone, variants in which a chemical moiety is bound to an N-linked or O-linked sugar chain, etc. Examples of biologically modified variants include variants obtained by post-translational modifications (e.g., N-linked or O-linked glycosylation, N-terminal or C-terminal processing, deamidation, isomerization of aspartic acid, oxidation of methionine, etc.), variants in which a methionine residue is added to the N-terminus by expression in a prokaryotic host cell, etc. Furthermore, antibodies labeled so as to be able to detect or isolate the antibodies or antigens according to the present disclosure, such as enzyme-labeled antibodies, fluorescent-labeled antibodies, and affinity-labeled antibodies, are also included in the meaning of the modified variants. Such modified variants of the antibodies according to the present disclosure are useful for improving the stability and blood retention of the antibodies, reducing antigenicity, detecting or isolating the antibodies or antigens, etc.

[0063] Furthermore, by regulating the modification (glycosylation, defucosylation, etc.) of the glycans linked to the antibodies according to the present disclosure, the antibody-dependent cell cytotoxic activity can be improved. Techniques for regulating the modification of the glycans of antibodies are known, such as those disclosed in WO 99 / 54342, WO 00 / 61739, WO 02 / 31140, WO 2007 / 133855, WO 2013 / 120066, etc. However, it is not limited thereto. The antibodies according to the present disclosure (preferably, anti-TROP2 or anti-HER2 antibodies) also include antibodies in which the modification of the glycans is regulated.

[0064] It is known that the lysine residue at the carboxyl terminus of the heavy chain of an antibody produced in mammalian cultured cells is deleted (Journal of Chromatography A, 705:129-134 (1995)), and that the two amino acid residues (glycine and lysine) at the carboxyl terminus of the heavy chain of an antibody produced in mammalian cultured cells are deleted, and that the proline residue newly located 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 (such as complement activation, antibody-dependent cell cytotoxicity, etc.) of the antibody. Therefore, the antibodies according to the present disclosure (preferably, anti-TROP2 or anti-HER2 antibodies) include antibodies and functional fragments of antibodies that have undergone such modifications, deletion mutants in which one or two amino acids are deleted at the carboxyl terminus of the heavy chain, mutants obtained by amidation of the deletion mutants (for example, heavy chains in which the carboxyl-terminal proline residue is amidated), and the like. The types of deletion mutants having a deletion at the carboxyl terminus of the heavy chain of the antibody according to the present disclosure are not limited to the above mutants as long as the antigen-binding affinity and effector functions are conserved. The two heavy chains constituting the antibody according to the present disclosure may be one selected from the group consisting of the full-length heavy chain and the deletion mutants described above, or a combination of two selected therefrom. The ratio of the amounts of the respective deletion mutants may be affected by the type of cultured mammalian cells producing the antibody according to the present disclosure and the culture conditions; however, antibodies in which one amino acid residue at the carboxyl terminus is deleted in both of the two heavy chains of the antibody according to the present disclosure can be exemplified as preferred ones.

[0065] Examples of the isotype of the antibody according to the present disclosure (preferably, anti-TROP2 or anti-HER2 antibody) include, for example, IgG (IgG1, IgG2, IgG3, IgG4), and preferably, IgG1 or IgG2.

[0066] The antibodies applicable to the production of the antibody-drug conjugates according to the present disclosure are not particularly limited to specific antigens. However, anti-TROP2 antibodies and anti-HER2 antibodies can be exemplified as preferred ones.

[0067] 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.

[0068] Examples of anti-TROP2 antibodies include, for example, hTINA1-H1L1 (International Publication No. 2015 / 098099), and preferably datopotamab may be mentioned.

[0069] In the present disclosure, the "anti-HER2 antibody" refers to an antibody that specifically binds to HER2 (human epidermal growth factor receptor type 2; ErbB-2), and preferably has the activity of internalizing into HER2-expressing cells by binding to HER2.

[0070] Examples of anti-HER2 antibodies include trastuzumab (U.S. Patent No. 5821337) and pertuzumab (International Publication No. 01 / 00245), and trastuzumab can be exemplified as a preferred one.

[0071] 3. Preparation of antibody-drug conjugate The drug-linker intermediate used in the production of the antibody-drug conjugate according to the present disclosure is represented by the following formula:

[0072]

Chemical formula

[0073] 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 International Publication No. WO 2014 / 057687, International Publication No. WO 2015 / 098099, International Publication No. WO 2015 / 115091, International Publication No. WO 2015 / 155998, International Publication No. WO 2019 / 044947, etc.

[0074] The antibody-drug conjugate used in the present disclosure can be manufactured by reacting the above drug-linker intermediate with an antibody having a thiol group (also called a sulfhydryl group) (preferably an anti-TROP2 antibody or an anti-HER2 antibody).

[0075] An antibody having a sulfhydryl group (preferably an anti-TROP2 or anti-HER2 antibody) can be obtained by a method 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 such as tris(2-carboxyethyl)phosphine hydrochloride (TCEP) at 0.3 to 3 molar equivalents with respect to the interchain disulfide in the antibody and reacting the antibody in a buffer solution containing a chelating agent such as ethylenediaminetetraacetic acid (EDTA), an antibody having a sulfhydryl group in which the interchain disulfide in the antibody is partially or completely reduced can be obtained.

[0076] Furthermore, by using 2 to 20 molar equivalents of a drug-linker intermediate with respect to an antibody having a sulfhydryl group (preferably an anti-TROP2 antibody or an anti-HER2 antibody), an antibody-drug conjugate in which 2 to 8 drug molecules are bound per antibody molecule can be prepared.

[0077] The average number of conjugated drug molecules per antibody (preferably an anti-TROP2 antibody or an anti-HER2 antibody) molecule of the produced antibody-drug conjugate can be determined, for example, by a method (UV method) calculated based on the measurement of UV absorbance at two wavelengths of 280 nm and 370 nm for the antibody-drug conjugate and its conjugation precursor, or by a method (HPLC method) calculated based on the quantification by HPLC measurement for the fragment obtained by treating the antibody-drug conjugate with a reducing agent.

[0078] The conjugation of an antibody (preferably an anti-TROP2 or anti-HER2 antibody) with a drug-linker intermediate and the calculation of the average number of conjugated drug molecules per antibody molecule of the antibody-drug conjugate can be carried out with reference to the descriptions in International Publication No. WO 2014 / 057687, International Publication No. WO 2015 / 098099, International Publication No. WO 2015 / 115091, International Publication No. WO 2015 / 155998, International Publication No. WO 2017 / 002776, International Publication No. WO 2018 / 212136, etc.

[0079] In the present invention, the term "anti-TROP2 antibody-drug conjugate" refers to an antibody-drug conjugate in which the antibody in the antibody-drug conjugate according to the present invention is an anti-TROP2 antibody.

[0080] The anti-TROP2 antibody preferably comprises a heavy chain comprising CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3 [= the amino acid sequence consisting of amino acid residues 50 to 54 of SEQ ID NO: 1], CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4 [= the amino acid sequence consisting of amino acid residues 69 to 85 of SEQ ID NO: 1], and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5 [= the amino acid sequence consisting of amino acid residues 118 to 129 of SEQ ID NO: 1], and a light chain comprising CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6 [= the amino acid sequence consisting of amino acid residues 44 to 54 of SEQ ID NO: 2], CDRL2 consisting of the amino acid sequence represented by SEQ ID NO: 7 [= the amino acid sequence consisting of amino acid residues 70 to 76 of SEQ ID NO: 2], and CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 8 [= the amino acid sequence consisting of amino acid residues 109 to 117 of SEQ ID NO: 2]. More preferably, the antibody comprises a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 [= the amino acid sequence consisting of amino acid residues 20 to 140 of SEQ ID NO: 1], and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 10 [= the amino acid sequence consisting of amino acid residues 21 to 129 of SEQ ID NO: 2]. Even more preferably, the antibody is one comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 12 [= the amino acid sequence consisting of amino acid residues 20 to 470 of SEQ ID NO: 1] and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 13 [= the amino acid sequence consisting of amino acid residues 21 to 234 of SEQ ID NO: 2], or an antibody comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 11 [= the amino acid sequence consisting of amino acid residues 20 to 469 of SEQ ID NO: 1] and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 13 [= SEQ ID NO: 2 amino acid residues 21 to 234].

[0081] The average number of conjugated drug-linkers per antibody molecule in the anti-TROP2 antibody-drug conjugate is preferably 2 to 8, more preferably 3 to 5, even more preferably 3.5 to 4.5, and even more preferably about 4.

[0082] The anti-TROP2 antibody-drug conjugate can be manufactured with reference to the descriptions in International Publication No. WO2015 / 098099 and International Publication No. WO2017 / 002776.

[0083] In a preferred embodiment, the anti-TROP2 antibody-drug conjugate is datopotamab deruxtecan (DS-1062).

[0084] In the present disclosure, the term "anti-HER2 antibody-drug conjugate" refers to an antibody-drug conjugate in which the antibody in the antibody-drug conjugate according to the present disclosure is an anti-HER2 antibody.

[0085] The anti-HER2 antibody is preferably an antibody comprising a heavy chain comprising CDRH1 consisting of the amino acid sequence consisting of amino acid residues 26 to 33 of SEQ ID NO: 14, CDRH2 consisting of the amino acid sequence consisting of amino acid residues 51 to 58 of SEQ ID NO: 14, and CDRH3 consisting of the amino acid sequence consisting of amino acid residues 97 to 109 of SEQ ID NO: 14, and a light chain comprising CDRL1 consisting of the amino acid sequence consisting of amino acid residues 27 to 32 of SEQ ID NO: 15, CDRL2 consisting of the amino acid sequence consisting of amino acid residues 50 to 52 of SEQ ID NO: 15, and CDRL3 consisting of the amino acid sequence consisting of amino acid residues 89 to 97 of SEQ ID NO: 15. More preferably, it is an antibody comprising a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence consisting of amino acid residues 1 to 120 of SEQ ID NO: 14 and a light chain comprising a light chain variable region consisting of the amino acid sequence consisting of amino acid residues 1 to 107 of SEQ ID NO: 15. Even more preferably, it is an antibody comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 14 and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 15, or an antibody comprising a heavy chain consisting of amino acid residues 1 to 449 of SEQ ID NO: 14 and a light chain consisting of the amino acid sequence consisting of all of amino acid residues 1 to 214 of SEQ ID NO: 15.

[0086] The average number of drug-linker units conjugated per antibody molecule in the anti-HER2 antibody-drug conjugate is preferably 2 to 8, more preferably 3 to 8, even more preferably 7 to 8, even more preferably 7.5 to 8, and even more preferably about 8.

[0087] The anti-HER2 antibody-drug conjugates used in the present disclosure can be manufactured with reference to the descriptions in WO 2015 / 115091 and the like.

[0088] In a preferred embodiment, the anti-HER2 antibody-drug conjugate is trastuzumab deruxtecan (DS-8201).

[0089] 4. Bispecific checkpoint inhibitors In the present disclosure, the term "bispecific checkpoint inhibitor" refers to a binding protein that is bispecific. As used herein, a bispecific binding protein has binding specificity for at least two independent antigens (or targets), or for different epitopes within the same antigen. Exemplary bispecific binding proteins may bind to two different epitopes within a target or to two different targets. Other such binding proteins may combine a first target binding site with a second binding site for a different target. In some embodiments, the binding protein is a bispecific antibody.

[0090] In some embodiments, the bispecific antibody provides additive and / or synergistic therapeutic effects resulting from simultaneous targeting of two antigens with the administration of a single manufactured molecule.

[0091] In some embodiments, the antibodies provided herein are monovalent bispecific antibodies (MBabs). The monovalent bispecific antibody scaffolds described herein provide an excellent platform for generating bispecific antibodies that, due to their monovalent nature, meet all of the benefits associated with bispecific antibodies while reducing the potential therapeutic risks described above. Additionally, the MBabs provided herein are readily expressed, stable, and likely to have low immunogenicity. As used herein, the term "monovalent bispecific," which may be abbreviated as "MBab," refers to a bispecific antibody in which each arm can specifically bind to a different target antigen, and for a given pair of different target antigens (A and B), the MBab can bind to one of each. In certain embodiments, the monovalent bispecific antibody can specifically bind to two independent antigens (or targets), or to two independent epitopes on the same antigen. Typically, a monovalent bispecific antibody comprises two different variable regions. In some embodiments, the binding affinities for the two independent antigens are approximately the same. In some embodiments, the binding affinities for the two independent antigens are different.

[0092] The bispecific checkpoint inhibitor is preferably a bispecific binding protein comprising a first binding domain that specifically binds to PD-1 and a second binding domain that specifically binds to CTLA-4 or TIGIT.

[0093] In a preferred embodiment, the bispecific binding protein is a) a first binding domain that specifically binds to PD-1, comprising a heavy chain variable domain comprising CDRH1 having the amino acid sequence of SEQ ID NO: 25, CDRH2 having the amino acid sequence of SEQ ID NO: 26, and CDRH3 having the amino acid sequence of SEQ ID NO: 27, and a light chain variable domain comprising CDRL1 having the amino acid sequence of SEQ ID NO: 28, CDRL2 having the amino acid sequence of SEQ ID NO: 29, and CDRL3 having the amino acid sequence of SEQ ID NO: 30, and b) A second binding domain that specifically binds to TIGIT, comprising a heavy chain variable domain comprising CDRH1 having the amino acid sequence of SEQ ID NO: 35, CDRH2 having the amino acid sequence of SEQ ID NO: 36, and CDRH3 having the amino acid sequence of SEQ ID NO: 37, and a light chain variable domain comprising CDRL1 having the amino acid sequence of SEQ ID NO: 38, CDRL2 having the amino acid sequence of SEQ ID NO: 39, and CDRL3 having the amino acid sequence of SEQ ID NO: 40; and a second binding domain.

[0094] More preferably, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 31 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 33. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 31. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a light chain variable domain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 33.

[0095] More preferably, the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 32 and a light chain having the amino acid sequence of SEQ ID NO: 34. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 32. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a light chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 34.

[0096] More preferably, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 43. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 41. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a light chain variable domain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 43.

[0097] More preferably, the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino sequence of SEQ ID NO: 42 and a light chain having the amino acid sequence of SEQ ID NO: 44. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 42. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a light chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 44.

[0098] In a particularly preferred embodiment, the bispecific binding protein is (a) a first binding domain that specifically binds to PD-1 and comprises a heavy chain having the amino acid sequence of SEQ ID NO: 32 and a light chain having the amino acid sequence of SEQ ID NO: 34; (b) a second binding domain that specifically binds to TIGIT and comprises a heavy chain having the amino sequence of SEQ ID NO: 42 and a light chain having the amino acid sequence of SEQ ID NO: 44, and is an antibody.

[0099] In another preferred embodiment, the bispecific binding protein is a) A first binding domain that specifically binds to PD-1, comprising a heavy chain variable domain comprising a CDRH1 having the amino acid sequence of SEQ ID NO: 52, a CDRH2 having the amino acid sequence of SEQ ID NO: 53, and a CDRH3 having the amino acid sequence of SEQ ID NO: 54, and a light chain variable domain comprising a CDRL1 having the amino acid sequence of SEQ ID NO: 49, a CDRL2 having the amino acid sequence of SEQ ID NO: 50, and a CDRL3 having the amino acid sequence of SEQ ID NO: 51; and the first binding domain. b) A second binding domain that specifically binds to CTLA-4, comprising a heavy chain variable domain comprising a CDRH1 having the amino acid sequence of SEQ ID NO: 58, a CDRH2 having the amino acid sequence of SEQ ID NO: 59, and a CDRH3 having the amino acid sequence of SEQ ID NO: 60, and a light chain variable domain comprising a CDRL1 having the amino acid sequence of SEQ ID NO: 55, a CDRL2 having the amino acid sequence of SEQ ID NO: 56, and a CDRL3 having the amino acid sequence of SEQ ID NO: 57; and the second binding domain.

[0100] More preferably, the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 46 and a light chain having the amino acid sequence of SEQ ID NO: 45. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 46. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a light chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 45.

[0101] More preferably, the second binding domain that specifically binds to CTLA-4 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 48 and a light chain having the amino acid sequence of SEQ ID NO: 47. In some embodiments, the second binding domain that specifically binds to CTLA-4 comprises a heavy chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 48. In some embodiments, the second binding domain that specifically binds to CTLA-4 comprises a light chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 47.

[0102] In a particularly preferred embodiment, the bispecific binding protein is (a) a first binding domain that specifically binds to PD-1 and comprises a heavy chain having the amino acid sequence of SEQ ID NO: 46 and a light chain having the amino acid sequence of SEQ ID NO: 45; and (b) a second binding domain that specifically binds to CTLA-4 and comprises a heavy chain having the amino acid sequence of SEQ ID NO: 48 and a light chain having the amino acid sequence of SEQ ID NO: 47.

[0103] In an even more preferred embodiment, the bispecific binding protein is MEDI5752. As used herein, the term "MEDI5752" or "brolucizumab" refers to an anti-PD-1 / CTLA-4 bispecific antibody comprising the light chain of SEQ ID NO: 45 and the heavy chain of SEQ ID NO: 46 (PD-1) and the light chain of SEQ ID NO: 47 and the heavy chain of SEQ ID NO: 48 (CTLA-4). MEDI5752 is disclosed in U.S. Patent No. 10,457,732, which is hereby incorporated by reference in its entirety.

[0104] In some embodiments, the light chain constant region of the bispecific binding protein is a kappa chain. In some embodiments, the light chain constant region is a lambda chain.

[0105] In some embodiments, when numbered by the EU index described in Kabat, the bispecific binding protein has the following amino acid residues: 221K, 221Y, 225E, 225K, 225W, 228P, 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235E, 235F, 236E, 237L, 237M, 237P, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 240I, 240A, 240T, 240M, 241W, 241L, 241Y, 241E, 241R, 243W, 243L, 243Y, 243R, 243Q, 244H, 245A, 247L, 247V, 247G, 250E, 250Q, 251F, 252L, 252Y, 254S, 254T, 255L, 256E, 256F, 256M, 257C, 257M, 257N, 262I, 262A, 262T, 262E, 263I, 263A, 263T, 263M, 264L, 264I, 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265A, 265G, 265N, 265Q, 265Y, 265F, 265V, 265I, 265L, 265H, 265T, 266I, 266A, 266T, 266M, 267Q, 267L, 268E, 269H, 269Y, 269F, 269R, 270E, 280A, 284M, 292P, 292L, 296E, 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H, 296G, 297S, 297D, 297E, 298A, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 305I, 308F, 313F, 316D, 318A, 318S, 320A, 320S, 322A, 322S, 325Q, 325L, 325I, 325D, 325E, 325A, 325T, 325V, 325H, 326A, 326D, 326E, 326G, 326M, 326V, 327G, 327W, 327N, 327L, 328S, 328M, 328D, 328E, 328N, 328Q, 328F, 328I, 328V, 328T, 328H, 328A, 329F,It comprises a mutant Fc region comprising at least one substitution selected from 329H, 329Q, 330K, 330G, 330T, 330C, 330L, 330Y, 330V, 330I, 330F, 330R, 330H, 331G, 331A, 331L, 331M, 331F, 331W, 331K, 331Q, 331E, 331S, 331V, 331I, 331C, 331Y, 331H, 331R, 331N, 331D, 331T, 332D, 332S, 332W, 332F, 332E, 332N, 332Q, 332T, 332H, 332Y, 332A, 333A, 333D, 333G, 333Q, 333S, 333V, 334A, 334E, 334H, 334L, 334M, 334Q, 334V, 334Y, 339T, 370E, 370N, 378D, 392T, 396L, 416G, 419H, 421K, 428L, 428F, 433K, 433L, 434A, 434W, 434Y, 436H, 440Y and 443W.,

[0106] In some embodiments, the mutant Fc region comprises one or more modifications at positions selected from 428 and 434 when numbered by the EU index shown in Kabat. In some embodiments, the mutant Fc region comprises one or more amino acid substitutions at positions selected from 428 and 434 when numbered by the EU index shown in Kabat. In some embodiments, the mutant Fc region comprises one or more amino acid substitutions selected from 428L, 428F, 434A, 434W, and 434Y.,

[0107] Engineering of the Fc region is widely used in the art to extend the half-life of therapeutic antibodies and protect them from in vivo degradation. In some embodiments, the Fc region of an IgG antibody or antigen-binding fragment can be modified to increase the affinity of the IgG molecule for the Fc receptor-neonatal (FcRn) that mediates IgG catabolism and protects the IgG molecule from degradation.,

[0108] In some embodiments, the antibody or antigen-binding fragment thereof comprises an Fc region engineered to improve the half-life.,

[0109] In some embodiments, the Fc region is aglycosylated. In some embodiments, the Fc region is deglycosylated. In some embodiments, the Fc region has reduced fucosylation or is not fucosylated.

[0110] In some embodiments, the Fc mutant antibody or its binding fragment has an increased binding affinity for FcRn.

[0111] The triple mutant (TM) L234F / L235E / P331S (according to the European Union numbering convention, Sazinsky et al. Proc Natl Acad Sci USA, 105:20167-20172 (2008)) can significantly reduce IgG effector function in the heavy chain constant region. In some embodiments, the antibody or its antigen-binding fragment comprises an Fc region having the L234F / L235E / P331S triple mutation (TM).

[0112] In some embodiments, the Fc mutant antibody or its binding fragment has reduced complement-dependent cytotoxicity (CDC) when administered in vivo. In some embodiments, the Fc mutant antibody or its binding fragment has reduced CDC compared to an antibody containing a wild-type Fc region or its binding variant. In some embodiments, the Fc mutant antibody or its binding fragment does not induce CDC when administered in vivo. In some embodiments, the Fc mutant antibody or its binding fragment causes a decrease in CDC when administered in vivo. In some embodiments, the Fc mutant antibody or its binding fragment having reduced or no CDC activity comprises a triple mutation (L234F / L235E / P331S) in the mutant Fc region.

[0113] In some embodiments, the Fc mutant antibody or its binding fragment has reduced antibody-dependent cell-mediated cytotoxicity (ADCC) when administered in vivo. In some embodiments, the Fc mutant antibody or its binding fragment has reduced ADCC compared to an antibody containing a wild-type Fc region or its binding variant. In some embodiments, the Fc mutant antibody or its binding fragment does not induce ADCC when administered in vivo. In some embodiments, the Fc mutant antibody or its binding fragment causes a decrease in ADCC when administered in vivo. In some embodiments, the Fc mutant antibody or its binding fragment having reduced ADCC activity or no ADCC activity comprises a triple mutation (L234F / L235E / P331S) in the mutant Fc region.

[0114] In some embodiments, an antibody or its binding fragment having reduced CDC activity has reduced toxicity when administered to a subject. In some embodiments, an antibody or its binding fragment having reduced ADCC activity has reduced toxicity when administered to a subject.

[0115] As used herein, the term "Fc domain" encompasses the native Fc and Fc variants and sequences defined above. Similar to Fc variants and native Fc molecules, the term "Fc domain" includes molecules in monomeric or multimeric form, whether digested from whole antibodies or produced by other means.

[0116] As used herein, the term "native Fc" refers to a molecule containing the sequence of a non-antigen-binding fragment that results from digestion of an antibody or is produced by other means, whether in monomeric or multimeric form, and can contain a hinge region. The original immunoglobulin source of the native Fc is preferably of human origin and can be any of the immunoglobulins. Native Fc molecules are composed of monomeric polypeptides that can be linked into dimeric or multimeric forms by covalent (i.e., disulfide bonds) and non-covalent bonds. The number of intermolecular disulfide bonds between the monomeric subunits of a native Fc molecule ranges from 1 to 4 depending on the class (e.g., IgG, IgA, and IgE) or subclass (e.g., IgG1, IgG2, IgG3, IgA1, and IgGA2). An example of native Fc is a disulfide-bonded dimer resulting from papain digestion of IgG. As used herein, the term "native Fc" is a general term for monomeric, dimeric, and multimeric forms.

[0117] In some embodiments, the Fc region is or comprises one or more domains that are Fc regions from IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD. In some embodiments, the antibody is an IgG1 antibody.

[0118] As used herein, the term "Fc variant" refers to a molecule or sequence that has been modified from native Fc but still contains the binding site for the salvage receptor, FcRn (neonatal Fc receptor). Exemplary Fc variants and their interactions with the salvage receptor are known in the art. Thus, the term "Fc variant" can include molecules or sequences that have been humanized from non-human native Fc. Further, native Fc can include regions that are removed or mutated to produce Fc variants in order to modify specific residues that provide structural features or biological activities not required for the binding proteins of the present disclosure. Thus, the term "Fc variant" refers to a molecule or sequence that lacks one or more native Fc sites or residues, or has one or more Fc sites or residues modified, that affect or are involved in (1) disulfide bond formation, (2) incompatibility with a selected host cell, (3) N-terminal heterogeneity upon expression in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to Fc receptors other than the salvage receptor, or (7) antibody-dependent cell cytotoxicity (ADCC).

[0119] To improve the yield of the binding protein, the CH3 domain can be modified by, for example, the "knob-into-hole" technology as described in detail with several exemplary molecules such as WO 96 / 027011, Ridgway et al., 1996, Protein Eng. 9:617-21, and Merchant et al., 1998, Nat. Biotechnol. 16:677-81. Specifically, the interaction surface of the two CH3 domains is changed to increase the heterodimerization of both heavy chains containing these two CH3 domains. Each of the two CH3 domains (of the two heavy chains) can be a "knob", while the other is a "hole". The introduction of a disulfide bridge further stabilizes the heterodimer (Merchant et al., 1998; Atwell et al., 1997, J. Mol. Biol. 270:26-35) and increases the yield.

[0120] In some embodiments, the binding protein has a "DuetMab" format. DuetMab has the following basic structure: An Fc region having a modified heavy chain, wherein the CH1 region of the modified heavy chain has substitutions from native cysteine to non-cysteine amino acids and from native non-cysteine amino acids to cysteine amino acids, an Fc region, a corresponding modified light chain, wherein the CL region of the modified light chain also has substitutions from native cysteine to non-cysteine amino acids and from native non-cysteine amino acids to cysteine amino acids, a second Fc region having a second heavy chain, and A second corresponding modified light chain, wherein the modified heavy chain is directly linked to the corresponding modified light chain, and on a separate target binding arm, the second heavy chain is directly linked to the second corresponding light chain, wherein the substituted cysteine of the modified heavy chain resulting from the substitution of a native non-cysteine amino acid to a cysteine amino acid and the substituted cysteine of the corresponding modified light chain resulting from the substitution of a native non-cysteine amino acid to a cysteine amino acid can form a disulfide bond. The disclosure regarding DuetMab exists, for example, in U.S. Patent No. 9,527,927, which is hereby incorporated by reference in its entirety.

[0121] As used herein, the term "KD" refers to the dissociation constant of the interaction between the binding protein of the present disclosure and the antigen target (KD = [A]×[B] / [AB]) and has units of moles per liter. The binding proteins of the present disclosure typically have a dissociation constant (KD) of 10-5 to 10-12 moles per liter or less, or 10-7 to 10-12 moles per liter or less, or 10-3 to 10-12 moles per liter, and / or a binding affinity of at least 107 M-1, or at least 108 M-1, or at least 109 M-1, or at least 1012 M-1. Any KD value greater than 10-4 moles per liter is generally considered to exhibit non-specific binding. Thus, the lower the KD value, the greater the affinity. In some embodiments, the binding proteins of the present disclosure bind to the desired antigen with an affinity of less than 500 nM, or less than 200 nM, or less than 10 nM, or less than 500 pM. High affinity or very strong binding is often associated with greater potency, but greater affinity does not necessarily mean greater potency.

[0122] The dissociation constant (KD) can be determined, for example, by surface plasmon resonance (SPR). Generally, surface plasmon resonance analysis measures the real-time binding interaction (both on-rate and off-rate) between a ligand (the target antigen on the biosensor matrix) and an analyte by surface plasmon resonance using, for example, a BIAcore® system (Pharmacia Biosensor; Piscataway, NJ). Surface plasmon analysis can also be performed by immobilizing the analyte and presenting the ligand. Specific binding of the binding proteins of the present disclosure to an antigen or antigenic determinant can also be determined in any suitable manner known in the art, including, for example, Scatchard analysis and / or competitive binding assays, such as radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), enzyme immunoassay (EIA), and sandwich competitive assays.

[0123] In one aspect, the equilibrium dissociation constant (KD) of the interaction between the bispecific binding protein described herein and human TIGIT is about 15 pM or less. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human TIGIT is about 9 pM or less. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human TIGIT is about 15, 14, 13, 12, 11, 10, 9, or 8 pM or less.

[0124] In one aspect, the KD of the interaction between the bispecific binding protein described herein and human TIGIT is from about 9 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human TIGIT is from about 10 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human TIGIT is from about 11 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human TIGIT is from about 12 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human TIGIT is from about 13 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human TIGIT is from about 14 pM to about 15 pM.

[0125] In one aspect, the KD of the interaction between the bispecific binding protein described herein and human PD-L1 is about 0.4 nM or less. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human PD-L1 is from about 0.2 nM to about 0.5 nM. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human PD-L1 is from about 0.3 nM to about 0.5 nM. In one aspect, the KD of the interaction between the bispecific binding protein described herein and human PD-L1 is from about 0.4 nM to about 0.5 nM.

[0126] 5. Combination of an Antibody-Drug Conjugate and a Bispecific Checkpoint Inhibitor In the first combination embodiment of the present disclosure, the antibody-drug conjugate combined with the bispecific checkpoint inhibitor is an antibody-drug conjugate in which the antibody is an anti-TROP2 antibody.

[0127] In one embodiment of the above first combination embodiment, the anti-TROP2 antibody comprises a heavy chain comprising CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3 [= amino acid residues 50-54 of SEQ ID NO: 1], CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4 [= amino acid residues 69-85 of SEQ ID NO: 1], and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5 [= amino acid residues 118-129 of SEQ ID NO: 1], and a light chain comprising CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6 [= amino acid residues 44-54 of SEQ ID NO: 2], CDRL2 consisting of the amino acid sequence represented by SEQ ID NO: 7 [= amino acid residues 70-76 of SEQ ID NO: 2], and CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 8 [= amino acid residues 109-117 of SEQ ID NO: 2]. In another embodiment of the above first combination embodiment, the anti-TROP2 antibody comprises a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 [= amino acid residues 20-140 of SEQ ID NO: 1], and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 10 [= amino acid residues twenty-one - 129 of SEQ ID NO: 2]. In another embodiment of the above first combination embodiment, the anti-TROP2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 12 [= amino acid residues 20-470 of SEQ ID NO: 1], and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 13 [= amino acid residues 21-234 of SEQ ID NO: 2]. In another embodiment of the above first combination embodiment, the anti-TROP2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 11 [= amino acid residues 20-469 of SEQ ID NO: 1], and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 13 [= amino acid residues 21-234 of SEQ ID NO: 2]. In another embodiment of the above first combination embodiment, the anti-TROP2 antibody is datopotamab deruxtecan (DS-1062).

[0128] In a second combination embodiment of the present disclosure, the antibody-drug conjugate combined with the bispecific checkpoint inhibitor is an antibody-drug conjugate in which the antibody is an anti-HER2 antibody.

[0129] In one embodiment of the second combination embodiment described above, the anti-HER2 antibody comprises a heavy chain comprising CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 16, CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 17, and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 18, and a light chain comprising CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 19, CDRL2 consisting of the amino acid sequence consisting of amino acid residues 1 to 3 of SEQ ID NO: 20, and CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 21. In another embodiment of the second combination embodiment described above, the anti-HER2 antibody comprises a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22 and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 23. In another embodiment of the second combination embodiment described above, the anti-HER2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 14 and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 15. In another embodiment of the second combination embodiment described above, the anti-HER2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 24 and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 15. In another embodiment of the second combination embodiment described above, the anti-HER2 antibody is trastuzumab deruxtecan (DS-8201).

[0130] In a particularly preferred embodiment of the above-described first combination embodiment, the anti-TROP2 antibody-drug conjugate is datopotamab deruxtecan (DS-1062), and the bispecific checkpoint inhibitor is an anti-PD-1 / TIGIT bispecific antibody in which the first binding domain that specifically binds to PD-1 includes a heavy chain having the amino acid sequence of SEQ ID NO: 32 and a light chain having the amino acid sequence of SEQ ID NO: 34, and the second binding domain that specifically binds to TIGIT includes a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 43.

[0131] In another particularly preferred embodiment of the above-described first combination embodiment, the anti-TROP2 antibody-drug conjugate is datopotamab deruxtecan (DS-1062), and the bispecific checkpoint inhibitor is an anti-PD-1 / CTLA-4 bispecific antibody in which the first binding domain that specifically binds to PD-1 includes a heavy chain having the amino acid sequence of SEQ ID NO: 46 and a light chain having the amino acid sequence of SEQ ID NO: 45, and the second binding domain that specifically binds to CTLA-4 includes a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 48 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 47.

[0132] In a particularly preferred embodiment of the above-described second combination embodiment, the anti-HER2 antibody-drug conjugate is trastuzumab deruxtecan (DS-8201), and the bispecific checkpoint inhibitor is an anti-PD-1 / TIGIT bispecific antibody in which the first binding domain that specifically binds to PD-1 includes a heavy chain having the amino acid sequence of SEQ ID NO: 32 and a light chain having the amino acid sequence of SEQ ID NO: 34, and the second binding domain that specifically binds to TIGIT includes a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 43.

[0133] In another particularly preferred embodiment of the second combination embodiment described above, the anti-HER2 antibody-drug conjugate is trastuzumab deruxtecan (DS-8201), and the bispecific checkpoint inhibitor is a heavy chain having a first binding domain that specifically binds to PD-1 and having the amino acid sequence of SEQ ID NO: 46 and a light chain having the amino acid sequence of SEQ ID NO: 45, and a second binding domain that specifically binds to CTLA-4 is a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 48 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 47, an anti-PD-1 / CTLA-4 bispecific antibody.

[0134] In some aspects of the combination embodiments described above, the antibody-drug conjugate and the bispecific checkpoint inhibitor are administered in further combination with one or more chemotherapeutic agents. In some aspects, the chemotherapeutic agent is carboplatin. In some aspects, the chemotherapeutic agent is a fluoropyrimidine (e.g., fluorouracil, 5-FU).

[0135] 6. Therapeutically combined uses and methods Pharmaceutical products, therapeutic uses and methods of administering in combination an antibody-drug conjugate (preferably an anti-TROP2 or anti-HER2 antibody-drug conjugate) according to the present disclosure and a bispecific checkpoint inhibitor are described below.

[0136] The pharmaceutical products, therapeutic uses and methods of the present disclosure may be characterized in that the antibody-drug conjugate and the bispecific checkpoint inhibitor are separately contained in different formulations as active ingredients and administered simultaneously or at different times, or the antibody-drug conjugate and the bispecific checkpoint inhibitor are contained in a single formulation as active ingredients and administered.

[0137] In the pharmaceutical products and treatment methods of the present disclosure, a single bispecific checkpoint inhibitor used in the present disclosure can be administered in combination with an antibody-drug conjugate, or two or more different bispecific checkpoint inhibitors can be administered in combination with an antibody-drug conjugate.

[0138] The pharmaceutical products and treatment methods of the present disclosure can be used for the treatment of cancer, preferably for the treatment of at least one cancer selected from the group consisting of 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 cancer, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumor, gastrointestinal stromal tumor, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, corpus cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, multiple myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer, and renal cell carcinoma.

[0139] The presence or absence of a tumor marker such as the HER2 or TROP2 tumor marker can be determined, for example, by collecting tumor tissue from a cancer patient to prepare a formalin-fixed paraffin-embedded (FFPE) specimen, and examining the gene product (protein) by immunohistochemistry (IHC), flow cytometry, Western blotting, etc., examining gene transcription by in situ hybridization (ISH), quantitative PCR (q-PCR), microarray analysis, etc., collecting cell-free circulating tumor DNA (ctDNA) from a cancer patient, and examining it by methods such as next-generation sequencing (NGS).

[0140] The pharmaceutical products and treatment methods of the present disclosure can be used for HER2-expressing cancer when they contain an anti-HER2 antibody-drug conjugate, and may be HER2-overexpressing cancer (high or medium), or HER2-low-expressing cancer.

[0141] In the present disclosure, the term "HER2-overexpressing cancer" is not particularly limited as long as it is recognized as HER2-overexpressing cancer by those skilled in the art. Preferred examples of HER2-overexpressing cancer include cancers that are given a score of 3+ for HER2 expression in the IHC method, and cancers that are given a score of 2+ for HER2 expression in the IHC method and are determined to be positive for HER2 expression in the in situ hybridization method (ISH). The in situ hybridization method of the present disclosure includes fluorescence in situ hybridization method (FISH) and dual-color in situ hybridization method (DISH).

[0142] In the present disclosure, the "HER2-low-expressing cancer" is not particularly limited as long as those skilled in the art recognize it as HER2-low-expressing cancer. Preferred examples of HER2-low-expressing cancer include cancers in which HER2 expression is 2+ in the IHC method and HER2 expression is determined to be negative in the in situ hybridization method, and cancers in which HER2 expression is 1+ in the IHC method.

[0143] The method of scoring the degree of HER2 expression by the IHC method or the method of determining positive or negative for HER2 expression by the in situ hybridization method is not particularly limited as long as it is recognized by those skilled in the art. For example, the methods described in the HER2 Testing Guideline, 4th Edition, Breast Cancer (developed by the HER2 Appropriate Use Committee of the Japanese Breast Cancer Pathology Society) can be mentioned.

[0144] The cancer may be HER2-overexpressing (high or medium) or low-expressing breast cancer, or triple-negative breast cancer, particularly with regard to the treatment of breast cancer, and / or may have a HER2 status score of IHC 3+, IHC 2+, IHC 1+ or IHC >0 and <1+.

[0145] In some embodiments, the method of the present disclosure includes identifying that the patient has a PD-L1 positive tumor prior to administration of the bispecific checkpoint inhibitor.

[0146] In some embodiments, the PD-L1 positive tumor comprises cells expressing at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 7%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or 100% PD-L1.

[0147] In some embodiments, PD-L1 expression is determined by receiving the results of an assay capable of determining PD-L1 expression. In some embodiments, the assay capable of determining PD-L1 expression is the Ventana PD-L1 (SP263) IHC assay, the 22C3 PharmDx assay, or the 28-8 PharmDx assay.

[0148] In some embodiments, the methods of the disclosure comprise administering an antibody-drug conjugate and an anti-PD-1 / CTLA-4 bispecific antibody to a patient whose tumor comprises less than 50% PD-L1-expressing cells. In some embodiments, the methods of the disclosure comprise administering an antibody-drug conjugate, an anti-PD-1 / CTLA-4 bispecific antibody, and carboplatin to a patient whose tumor comprises less than 50% PD-L1-expressing cells.

[0149] In some embodiments, the methods of the disclosure comprise administering an antibody-drug conjugate and an anti-PD-1 / TIGIT bispecific antibody to a patient whose tumor comprises at least 50% PD-L1-expressing cells. In some embodiments, the methods of the disclosure comprise administering an antibody-drug conjugate, an anti-PD-1 / TIGIT bispecific antibody, and carboplatin to a patient whose tumor comprises less than 50% PD-L1-expressing cells.

[0150] The pharmaceutical products and treatment methods of the disclosure can preferably be used in mammals, more preferably in humans.

[0151] The antitumor effects of the pharmaceutical products and treatment methods of the present disclosure can be confirmed by transplanting cancer cells into a subject animal to create a model and measuring the reduction in tumor volume or the life-prolonging effect by applying the pharmaceutical products and treatment methods of the present disclosure. Next, the effect of the combined use of the antibody-drug conjugate and the bispecific checkpoint inhibitor used in the present disclosure can be confirmed by comparing the antitumor effect by single administration of the antibody-drug conjugate used in the present disclosure with that of the bispecific checkpoint inhibitor.

[0152] The antitumor effects of the pharmaceutical products and treatment methods of the present disclosure can be confirmed in clinical trials using any of the evaluation methods according to Response Evaluation Criteria in Solid Tumors (RECIST), WHO evaluation method, Macdonald evaluation method, body weight measurement, and other approaches, 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.

[0153] By using the above methods, it can be confirmed that the antitumor effects of the pharmaceutical products and treatment methods of the present disclosure are superior to those of the existing pharmaceutical products and treatment methods for cancer treatment.

[0154] The pharmaceutical products and treatment methods of the present disclosure can delay the occurrence of cancer cells, inhibit their growth, and further kill cancer cells. Due to these effects, cancer patients can be liberated from cancer-related symptoms, the quality of life (QOL) of cancer patients can be improved, and the life of cancer patients can be maintained to obtain a therapeutic effect. Even if the pharmaceutical products and treatment methods of the present disclosure do not achieve the killing of cancer cells, by inhibiting or controlling the growth of cancer cells, a higher QOL of cancer patients can be achieved while achieving a longer survival.

[0155] In addition to its application as a systemic therapy for patients, the pharmaceutical product of the present disclosure can be expected to exert a therapeutic effect by local application to cancer tissues.

[0156] The pharmaceutical product and treatment method of the present disclosure, in another aspect, 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 include administering a therapeutically effective amount of the pharmaceutical product to a subject in need thereof, either simultaneously with or sequentially to ionizing radiation or other chemotherapeutic agents.

[0157] The pharmaceutical product and treatment method of the present disclosure can be used as adjuvant chemotherapy in combination with surgery. The pharmaceutical product of the present disclosure may be administered for the purpose of reducing tumor size before surgery (referred to as neoadjuvant chemotherapy or preoperative adjuvant therapy), or may be administered for the purpose of preventing tumor recurrence after surgery (referred to as postoperative adjuvant chemotherapy or adjuvant therapy).

[0158] In some embodiments, the cancer cells may have a BRCA1 and / or BRCA2-deficient phenotype, i.e., BRCA1 and / or BRCA2 activity is reduced or absent in the cancer cells. Cancer cells having this phenotype may be deficient in BRCA1 and / or BRCA2, i.e., the expression and / or activity of BRCA1 and / or BRCA2 may be reduced or absent in the cancer cells, for example, by a mutation or polymorphism in the coding nucleic acid, or by amplification, mutation or polymorphism in a gene encoding a regulatory factor, such as the EMSY gene encoding a BRCA2 regulatory factor (Hughes-Davies, et al., Cell, 115, 523-535). BRCA1 and BRCA2 are known tumor suppressors whose wild-type alleles are frequently 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 between BRCA1 and / or BRCA2 mutations and 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 known to be associated with breast and ovarian cancers. Carriers of mutations in BRCA1 and / or BRCA2 are also at high risk for certain cancers, including breast, ovarian, pancreatic, prostate, blood, gastrointestinal and lung cancers. In some embodiments, the individual is heterozygous for one or more mutations, e.g., mutations and polymorphisms, in BRCA1 and / or BRCA2 or its regulatory factors.The detection of mutations in BRCA1 and BRCA2 is well known in the art and is described, for example, in European Patent No. 699 754, European Patent No. 705 903, 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 the amplification of the BRCA2 binding factor EMSY is described in Hughes-Davies, et al., Cell, 115, 523-535).

[0159] Cancer-related mutations and polymorphisms can be detected at the nucleic acid level by detecting the presence of mutant nucleic acid sequences or at the protein level by detecting the presence of mutant (i.e., mutant or allelic variant) polypeptides.

[0160] The pharmaceutical products of the present disclosure can be administered containing at least one pharmaceutically suitable component. The pharmaceutically suitable component can be appropriately selected and applied from formulation additives generally used in the art according to the dosage, administration concentration, etc. of the antibody-drug conjugate and bispecific checkpoint inhibitor used in the present disclosure. The antibody-drug conjugate used in the present disclosure can be administered as a pharmaceutical product containing, for example, a buffer such as 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 lyophilized injection, and still more preferably as a lyophilized injection. When the pharmaceutical product containing the antibody-drug conjugate used in the present disclosure is an aqueous injection, the aqueous injection can preferably be administered as an intravenous injection after being diluted with a suitable diluent. Examples of the diluent include, for example, a dextrose solution and physiological saline, preferably a dextrose solution, and more preferably a 5% dextrose solution. When the pharmaceutical product of the present disclosure is a lyophilized injection, the required amount of the lyophilized injection pre-dissolved in distilled water for injection can preferably be administered as an intravenous injection after being diluted with a suitable diluent. Examples of the diluent include, for example, a dextrose solution and physiological saline, preferably a dextrose solution, and more preferably a 5% dextrose solution.

[0161] Examples of the administration routes applicable to the administration of the pharmaceutical products of the present disclosure include intravenous, intradermal, subcutaneous, intramuscular and intraperitoneal routes, with the intravenous route being preferred.

[0162] The size of the dose required for the therapeutic treatment of a particular disease state necessarily varies depending on the subject being treated, the route of administration, and the severity of the disease being treated. Further information regarding the route of administration and dosing schedule can be found, for example, in Chapter 25.3 of Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.

[0163] The anti-TROP2 antibody-drug conjugate used in the present disclosure can be administered to humans once at intervals of 1 to 180 days, preferably once a week, once every two weeks, once every three weeks, or once every four weeks, and more preferably once every three weeks. In addition, the antibody-drug conjugate used in the present invention can be administered at a dose of about 0.001 to 100 mg / kg, preferably at a dose of 0.8 to 12.4 mg / kg. For example, the anti-TROP2 antibody-drug conjugate can be administered once every three weeks at a dose of 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, preferably at a dose of 4.0 or 6.0 mg / kg once every three weeks.

[0164] The anti-HER2 antibody-drug conjugate used in the present disclosure can be administered to humans at intervals of 1 to 180 days, preferably at intervals of 1 week, 2 weeks, 3 weeks, or 4 weeks, and more preferably at intervals of 3 weeks. The anti-HER2 antibody-drug conjugate used in the present disclosure can be administered at a dose of about 0.001 to 100 mg / kg per administration, preferably at a dose of 0.8 to 12.4 mg / kg per administration. For example, the anti-HER2 antibody-drug conjugate can be administered once every 3 weeks at a dose of 0.8 mg / kg, 1.6 mg / kg, 3.2 mg / kg, 5.4 mg / kg, 6.4 mg / kg, 7.4 mg / kg, or 8 mg / kg, preferably at a dose of 5.4 mg / kg or 6.4 mg / kg once every 3 weeks.

[0165] The bispecific checkpoint inhibitor can be administered at an appropriate dose by any suitable route of administration.

[0166] In some embodiments, MEDI5752 or an antigen-binding fragment thereof is administered to a subject at a dose of about 100 mg to about 1500 mg. In some embodiments, the dose for administration is about 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, about 1000 mg, about 1010 mg, about 1020 mg, about 1030 mg, about 1040 mg, about 1050 mg, about 1060 mg, about 1070 mg, about 1080 mg, about 1090 mg, about 1100 mg, about 1120 mg, about 1130 mg, about 1140 mg, about 1150 mg, about 1160 mg, about 1170 mg, about 1180 mg, about 1190 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg.

[0167] In some embodiments, the dose of MEDI5752 or an antigen-binding fragment thereof is administered to a subject once per treatment cycle. In some embodiments, the treatment cycle is three weeks. In some embodiments, the dose of MEDI5752 or an antigen-binding fragment thereof is administered every three weeks for about 12 months, about 24 months, about 36 months, or about 48 months.

[0168] In some embodiments, the bispecific antibody or an antigen-binding fragment thereof is administered in combination with one or more chemotherapeutic agents. In some embodiments, the chemotherapeutic agent is carboplatin. In some embodiments, the chemotherapeutic agent is a fluoropyrimidine (e.g., fluorouracil, 5-FU). In some embodiments, the chemotherapeutic agent is pemetrexed. In some embodiments, the chemotherapeutic agent is axitinib.

[0169] In some aspects, bispecific checkpoint inhibitors such as the binding proteins disclosed herein can be formulated as pharmaceutical compositions together with pharmaceutically acceptable carriers, excipients, or stabilizers. In certain aspects, such pharmaceutical compositions are suitable for administration to humans or non-human animals via any one or more routes of administration using methods known in the art. The term "pharmaceutically acceptable carrier" means one or more non-toxic substances that do not interfere with the biological activity effectiveness of the active ingredient. Such preparations may conventionally contain salts, buffers, preservatives, compatible carriers, and optionally other therapeutic agents. Such pharmaceutically acceptable preparations 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 may be utilized 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, salt-forming counterions such as sodium, and the like. These and additional known pharmaceutical carriers, excipients, and / or additives suitable for use in the formulations described herein are known in the art, as exemplified, for example, in "Remington: The Science & Practice of Pharmacy", 21st Edition, Lippincott Williams & Wilkins, (2005), and "Physician’s Desk Reference", 60th Edition, Medical Economics, Montvale, N.J. (2005). A pharmaceutically acceptable carrier suitable for the desired or required mode of administration, solubility, and / or stability can be selected.

[0170] In some embodiments, the therapeutic composition can be formulated for administration by a particular route, such as oral, nasal, pulmonary, topical (including buccal and sublingual), rectal, vaginal, and / or parenteral. As used herein, the terms “parenteral administration” and “administered parenterally” refer to a mode of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion. Formulations of the 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 as may be required (see, e.g., 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).

[0171] The formulation can be provided in unit dosage form and can be prepared by any method known in the pharmaceutical art. The actual dosage level of the active ingredient in the pharmaceutical composition of the present disclosure is not toxic to the patient and can be varied to obtain an amount of the active ingredient (e.g., a "therapeutically effective amount") effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration. The dosage level selected will depend on various pharmacokinetic factors such as 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, compounds and / or substances used in combination with the particular composition employed, the age, sex, weight, condition, general health, and prior medical history of the patient being treated, and like factors well known in the medical arts. These dosages can be administered daily, weekly, biweekly, monthly, or less frequently (e.g., semi-annually), depending on the dosage, the method of administration, the disorder or condition 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 noted above, any commonly used dosing regimen (e.g., administering 1-10 mg / kg once a day or twice a week by injection or infusion) can be adapted and appropriate in methods for treating human cancer patients.

Example

[0172] Hereinafter, the present disclosure will be specifically described based on examples. However, the present disclosure is not limited thereto. Furthermore, this should in no way be construed as limiting.

[0173] Example 1A: Preparation of Anti-TROP2 Antibody-Drug Conjugate According to the production methods described in International Publication No. WO 2015 / 098099 and International Publication No. WO 2017 / 002776, using an anti-TROP2 antibody (an antibody comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 12 [= amino acid residues 20 to 470 of SEQ ID NO: 1] and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 13 [= amino acid residues 21 to 234 of SEQ ID NO: 2]), a drug-linker has the following formula:

[0174] [Chemical formula] Anti-TROP2 antibody-drug conjugate represented by (In the formula, A represents the connection position to the antibody and is conjugated to the anti-TROP2 antibody via a thioether bond) was prepared (DS-1062: datopotamab deruxtecan). The DAR of the antibody-drug conjugate is ~4.

[0175] Example 1B: Preparation of anti-HER2 antibody-drug conjugate According to the production method described in International Publication No. 2015 / 115091, using an anti-HER2 antibody (an antibody comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 24 (amino acid residues 1 to 449 of SEQ ID NO: 14) and a light chain consisting of the amino acid sequence consisting of all of amino acid residues 1 to 214 of SEQ ID NO: 15), a drug-linker has the following formula:

[0176] [Chemical formula] Anti-HER2 antibody-drug conjugate represented by (In the formula, A represents the connection position to the antibody and is conjugated to the anti-HER2 antibody via a thioether bond) was prepared (DS-8201: trastuzumab deruxtecan). The DAR of the antibody-drug conjugate is 7.7 or 7.8.

[0177] Example 2A: Anti-PD-1 / TIGIT bispecific antibody (bispecific checkpoint inhibitor) A first binding domain that specifically binds to PD-1, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 32 and a light chain having the amino acid sequence of SEQ ID NO: 34, and a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 42 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 44, to produce an anti-PD-1 / TIGIT bispecific antibody (monovalent, humanized, IgG1 monoclonal) having a second binding domain that specifically binds to TIGIT. The antibody is engineered to have a triple mutation L234F / L235E / P331S within its Fc domain to reduce Fc-mediated effector function.

[0178] Example 2B: Anti-PD-1 / CTLA-4 Bispecific Antibody (Bispecific Checkpoint Inhibitor) Produce the anti-PD-1 / CTLA-4 bispecific antibody MEDI5752 disclosed in U.S. Patent No. 10,457,732. MEDI5752 has a first binding domain that specifically binds to PD-1, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 46 and a light chain having the amino acid sequence of SEQ ID NO: 45, and a second binding domain that specifically binds to CTLA-4, comprising a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 48 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 47.

[0179] Example 3: Antitumor Test Combination of Antibody-Drug Conjugate DS-8201 and Anti-PD-L1 + Anti-CTLA-4 or Anti-PD-1 / TIGIT DuetMab (i) Method: Tumor Model Female BALB / c mice, 7 - 9 weeks old, were purchased from Envigo and acclimated for at least 7 days before entering the study. For tumor transplantation, the right flank of the mice was shaved, and 100 μL of cells containing 5 × 10 6 cells of EMT6 human HER2 (hHER2) were injected subcutaneously. Tumor volume was measured three times a week using an electronic caliper and calculated using the formula (width 2 × length) / 2.

[0180] Seven days after cell transplantation, when the tumor was approximately 170 mm3 Once the limit was reached, tumors of the same size were randomly assigned to the treatment groups as shown in Table 1. When the humane welfare limit regarding tumor volume (average diameter 15 mm) or tumor condition (ulceration of the skin over the tumor) was reached, the mice were euthanized.

[0181] Humanized female NSG mice were transplanted with CD34+ cord blood stem cells at the Jackson Laboratory using three different donors. Approximately 15 weeks after engraftment, 1e6 Caki-1 cells were subcutaneously injected into the right flank. Tumor volume was measured twice a week using calipers and calculated using the formula (width 2 × length) / 2. When the tumors reached an average volume of 115 mm 3 the mice were randomized to treatment groups as shown in Table 2. The mice were randomized by both tumor volume and cord blood donor to ensure three donors per group.

[0182]

Table 1

[0183]

Table 2

[0184] Formulation Formulation of 10 mg / kg DS-8201 The dosing solution of DS-8201 (HA306) was prepared by diluting the DS-8201 stock (20.1 mg / mL) in ABS buffer (10 mM sodium acetate, 10 mM acetic acid, 5% sorbitol) to 2.5 mg / mL and then administered by IV injection at a dosing volume of 4 mL / kg.

[0185] Formulation of 10 mg / kg anti-PD-L1 The dosing solution of anti-PD-L1 (clone 80, SP21-095, IgG1, D265A) was prepared by diluting the stock (11.2 mg / mL) in PBS to 1 mg / mL and then administered by IP injection at a dosing volume of 10 mL / kg.

[0186] Co-formulation of anti-PD-L1 and anti-CTLA-4 at 10 mg / kg The dosing solutions of anti-PD-L1 (clone 80, SP21-095, IgG1, D265A) and anti-CTLA-4 (SP20-103, mouse IgG1) were prepared by diluting the stocks (anti-PD-L1 - 11.2 mg / mL; anti-CTLA-4 antibody - 10 mg / mL) and co-formulating in PBS at 1 mg / mL for each agent, and then administered by IP injection at a dosing volume of 10 mL / kg.

[0187] Formulation of anti-PD-1 / TIGIT DuetMab The dosing solution of anti-PD-1 / TIGIT DuetMab (mIgG2a LALA-PG) was prepared by diluting the stock in PBS (C428223DEC21EO, 2.7 mg / mL) to 1 mg / mL, and then administered by IP injection at a dosing volume of 10 mL / kg.

[0188] Formulation of MEDI5752 The dosing solution of MEDI5752 was prepared by diluting the stock in PBS (lot number ML00669-50, 63.41 mg / mL) to 2 mg / mL, and then administered by IP injection at a dosing volume of 5 mL / kg.

[0189] Fine needle aspiration (FNA) sampling Mice were anesthetized with isoflurane and placed on a rotating anesthesia platform. A 25-gauge needle was attached to a 1 mL syringe filled with 0.9 mL of cold RPMI medium. The needle was inserted horizontally into the tumor and rotated to remove tissue. Negative pressure was generated by pulling back the syringe plunger approximately 100 μL to draw cells into the medium. Approximately 200 μL of the medium containing the cells was then flushed from the syringe into a 1.5 mL Eppendorf tube. This was repeated 4 more times at different sites around the entire tumor to maximize the representativeness of the sampling, and the tube was immediately placed on ice.

[0190] Flow cytometry staining Cells were stained with a fixable viability dye (Thermo Fisher), blocked with an antibody against CD16 / CD32 (eBioscience), and then stained with fluorescent-conjugated antibodies in flow cytometry staining buffer (2% bovine serum albumin, 0.1% sodium azide, 2 mM EDTA) containing 50% Brilliant Stain Buffer (BD Biosciences). Intracellular staining was performed using the FoxP3 / Transcription Factor Staining Buffer Set (eBioscience), and the cells were fixed in 3.7% formaldehyde. Cells were acquired in flow cytometry staining buffer on a BD FACSymphony flow cytometer (BD Biosciences) and analyzed using FlowJo (TreeStar). Samples containing less than 1000 CD45 + cells were excluded from the analysis due to low event numbers.

[0191] Statistics The tumor growth rate for each mouse was calculated based on fitting the growth curve (days 0 to 15 after treatment initiation) of each tumor to an exponential model log10(tumor volume) = a + b·time + error, where a and b are parameters corresponding to log initial volume and growth rate, respectively. The statistical significance of the difference in tumor growth rates between groups was evaluated using the Mann–Whitney U test. Box-and-whisker plots show the median and 25 番目 –75 番目 th quartiles, and whiskers show the minimum and maximum values.

[0192] For flow cytometry data of cell frequencies, the data were processed using beta regression as a fixed effect. p-values were reported as follows: p ≥ 0.05 (ns, not significant); * , p < 0.05; ** , p < 0.01; *** , p < 0.001; **** , p < 0.0001.

[0193] (ii) Tests: Mice bearing EMT6 hHER2 tumors were treated with a combination of DS-8201 and Immuno-Oncology (IO) agents according to the dosing schedule outlined in Table 1, i.e., with 10 mg / kg of DS-8201 alone or in combination with 10 mg / kg of anti-PD-L1, 10 mg / kg of anti-PD-L1 + 10 mg / kg of anti-CTLA-4, and 10 mg / kg of anti-PD-1 / TIGIT DuetMab. The results are shown in FIGS. 59 and 60 and Tables 3 and 4.

[0194] FNA tumor samples were collected 8 days after the start of treatment from EMT6 hHER2 tumor-bearing mice treated with a combination of DS-8201 alone or in combination with 10 mg / kg of anti-PD-1 / TIGIT DuetMab according to the dosing schedule outlined in Table 1, and pharmacodynamic changes were evaluated by flow cytometry. Samples with less than 1000 CD45+ cells were excluded from the analysis (n = 7 - 9 FNA samples / group). T cells = CD45+CD3+ cells. Tregs = CD45+CD3+CD4+FoxP3+ cells. NK cells = CD45+CD3-NKp46+ cells. The results are shown in FIG. 61. In a similar study, tumor samples were collected 10 days after the start of treatment from mice bearing EMT6 hHER2 tumors treated with vehicle or DS-8201 alone, and pharmacodynamic changes in CD8+ T cell CTLA-4 expression were evaluated by flow cytometry (n = 8 samples / group). The results are shown in FIG. 62.

[0195] In a separate study, mice bearing Caki-1 tumors were treated with a combination of DS-8201 and / or the IO agent MEDI5752 according to the dosing schedule outlined in Table 2. The results are shown in FIG. 63.

[0196] Peripheral blood was sampled on day 22 after the start of treatment in the Caki-1 study and analyzed via flow cytometry for human-specific T cell activation markers (n = 5 samples / group). The results are shown in FIG. 64.

[0197] (iii) Results: Figures 59 and 60 show the antitumor efficacy and tumor growth rate of the combination of DS-8201 and IO agents in EMT6 hHER2 tumor-bearing mice. In Figure 59, Graph A shows the average change in tumor volume over time. The data shown are mean ± SEM (n = 10 mice / treatment group). In Figure 59, Graph B shows the change in tumor volume over time for individual mice (CR = complete response). In Figure 60, the data shown are tumor growth rate (n = 10 mice / treatment group).

[0198] Monotherapy with DS-8201 significantly delayed tumor growth (53.2% growth rate inhibition, p = 0.0002), similar to anti-PD-L1 (46.2% growth rate inhibition, p = 0.0185), anti-PD-L1 + anti-CTLA-4 (86.2% growth rate inhibition, p = 0.0355), and anti-PD-1 / TIGIT DuetMab (120.3% growth rate inhibition, p < 0.0001) (Figure 59, Graph A, Figure 60, Table 3).

[0199]

Table 3

[0200] Tumor growth inhibition was further enhanced when DS-8201 was combined with anti-PD-L1 (117.4% growth rate inhibition, p = 0.0007), anti-PD-L1 + anti-CTLA-4 (253.5% growth rate inhibition, p = 0.0002), and anti-PD-1 TIGIT DuetMab (273.6% growth rate inhibition, p < 0.0001) compared to DS-8201 monotherapy (Figure 59, Graph A, Table 4). The combinations of DS-8201 with anti-PD-L1 + anti-CTLA-4 and anti-PD-1 / TIGIT DuetMab resulted in complete responses in 80% and 60% of the animals, respectively, while the combination with anti-PD-L1 resulted in a complete response in only 10% of the animals (Graph B in Figure 59, Table 4).

[0201]

Table 4

[0202] Regarding the pharmacodynamic changes, FNA tumor samples collected 8 days after the start of DS-8201 treatment revealed an increase in the tumor proportion of total immune cells (CD45+), Tregs, and NK cells, as well as an increase in the proportion of T cells expressing PD-1, Ki67, and TIGIT. The combination of DS-8201 and anti-PD-1 / TIGIT DuetMab treatment further increased the tumor T cell (% survival) and NK cell (% CD45+) content compared to monotherapy, and also increased the proportion of CD8+ T cells (% CD3+) (Figure 61). In a similar study, DS-8201 monotherapy significantly increased the expression of CTLA-4 on tumor CD8+ T cells (Figure 62).

[0203] In a CD34+ hematopoietic stem cell transplant humanized mouse model bearing renal Caki-1 tumors, the combination of DS-8201 and MEDI5752 resulted in greater tumor growth inhibition than either monotherapy alone and was significantly better than DS-8201 (Figure 63). Compared to the DS-8201 treatment group, mice receiving MEDI5752 monotherapy showed substantial weight loss, which led to the early termination of this treatment group on day 50 after tumor transplantation. Interestingly, these effects were not observed in the group receiving MEDI5752 in combination with DS-8201.

[0204] Peripheral blood flow analysis was performed on day 22 after treatment in the Caki-1 study, revealing an increase in proliferating CD4+ and CD8+ T cells and an increase in granzyme B+ CD8 and ICOS+ CD4 cells. These effects were observed with monotherapy MEDI5752 and maintained in the combination treatment group (Figure 64).

[0205] The foregoing specification is considered to be sufficient to enable those skilled in the art to practice the embodiments. The foregoing description and examples detail particular embodiments and illustrate the best mode contemplated by the inventors. However, it will be understood that, however detailed the foregoing appears in text, the embodiments may be practiced in many ways and the claims include any equivalents thereof.

[0206] Free text of the sequence listing Amino acid sequence of the heavy chain of SEQ ID NO: 1 - anti-TROP2 antibody Amino acid sequence of the light chain of SEQ ID NO: 2 - anti-TROP2 antibody Amino acid sequence of heavy chain CDRH1 [= amino acid residues 50 - 54 of SEQ ID NO: 1] Amino acid sequence of heavy chain CDRH2 [= amino acid residues 69 - 85 of SEQ ID NO: 1] Amino acid sequence of heavy chain CDRH3 [= amino acid residues 118 - 129 of SEQ ID NO: 1] Amino acid sequence of light chain CDRL1 [= amino acid residues 44 - 54 of SEQ ID NO: 2] Amino acid sequence of light chain CDRL2 [= amino acid residues 70 - 76 of SEQ ID NO: 2] Amino acid sequence of light chain CDRL3 [= amino acid residues 109 - 117 of SEQ ID NO: 2] Amino acid sequence of the heavy chain variable region [= amino acid residues 20 - 140 of SEQ ID NO: 1] Amino acid sequence of the light chain variable region [= amino acid residues 21 - 129 of SEQ ID NO: 2] Amino acid sequence of the heavy chain [= amino acid residues 20 - 469 of SEQ ID NO: 1] Amino acid sequence of the heavy chain [= amino acid residues 20 - 470 of SEQ ID NO: 1] Amino acid sequence of the light chain [= amino acid residues 21 - 234 of SEQ ID NO: 2] Amino acid sequence of the heavy chain of SEQ ID NO: 14 - anti-HER2 antibody Amino acid sequence of the light chain of SEQ ID NO: 15 - anti-HER2 antibody Amino acid sequence of heavy chain CDRH1 [= amino acid residues 26 - 33 of SEQ ID NO: 14] Amino acid sequence of heavy chain CDRH2 [= amino acid residues 51 - 58 of SEQ ID NO: 14] Amino acid sequence of heavy chain CDRH3 [= amino acid residues 97 - 109 of SEQ ID NO: 14] Amino acid sequence of light chain CDRL1 [= amino acid residues 27 - 32 of SEQ ID NO: 15] Amino acid sequence containing the amino acid sequence of SEQ ID NO: 20 - light chain CDRL2 (SAS) [= amino acid residues 50 - 56 of SEQ ID NO: 15] Amino acid sequence of SEQ ID NO: 21 - light chain CDRL3 [= amino acid residues 89 - 97 of SEQ ID NO: 15] Amino acid sequence of SEQ ID NO: 22 - heavy chain variable region [= amino acid residues 1 - 120 of SEQ ID NO: 14] Amino acid sequence of SEQ ID NO: 23 - light chain variable region [= amino acid residues 1 - 107 of SEQ ID NO: 15] Amino acid sequence of SEQ ID NO: 24 - heavy chain [= amino acid residues 1 - 449 of SEQ ID NO: 14] Amino acid sequence of SEQ ID NO: 25 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 heavy chain CDRH1 Amino acid sequence of SEQ ID NO: 26 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 heavy chain CDRH2 Amino acid sequence of SEQ ID NO: 27 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 heavy chain CDRH3 Amino acid sequence of SEQ ID NO: 28 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 light chain CDRL1 Amino acid sequence of SEQ ID NO: 29 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 light chain CDRL2 Amino acid sequence of SEQ ID NO: 30 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 light chain CDRL3 Amino acid sequence of SEQ ID NO: 31 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 heavy chain variable region Amino acid sequence of SEQ ID NO: 32 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 heavy chain Amino acid sequence of SEQ ID NO: 33 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 light chain variable region Amino acid sequence of SEQ ID NO: 34 - anti - PD - 1 / TIGIT bispecific antibody, anti - PD1 light chain Amino acid sequence of SEQ ID NO: 35 - anti - PD - 1 / TIGIT bispecific antibody, anti - TIGIT heavy chain CDRH1 Amino acid sequence of SEQ ID NO: 36 - anti - PD - 1 / TIGIT bispecific antibody, anti - TIGIT heavy chain CDRH2 Amino acid sequence of the anti-TIGIT heavy chain CDRH3 of the array number 37 - anti-PD-1 / TIGIT bispecific antibody Amino acid sequence of the anti-TIGIT light chain CDRL1 of the array number 38 - anti-PD-1 / TIGIT bispecific antibody Amino acid sequence of the anti-TIGIT light chain CDRL2 of the array number 39 - anti-PD-1 / TIGIT bispecific antibody Amino acid sequence of the anti-TIGIT light chain CDRL3 of the array number 40 - anti-PD-1 / TIGIT bispecific antibody Amino acid sequence of the anti-TIGIT heavy chain variable region of the array number 41 - anti-PD-1 / TIGIT bispecific antibody Amino acid sequence of the anti-TIGIT heavy chain of the array number 42 - anti-PD-1 / TIGIT bispecific antibody Amino acid sequence of the anti-TIGIT light chain variable region of the array number 43 - anti-PD-1 / TIGIT bispecific antibody Amino acid sequence of the anti-TIGIT light chain of the array number 44 - anti-PD-1 / TIGIT bispecific antibody Amino acid sequence of the anti-PD1 light chain of the array number 45 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-PD1 heavy chain of the array number 46 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-CTLA-4 light chain of the array number 47 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-CTLA-4 heavy chain of the array number 48 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-PD1 light chain CDRL1 of the array number 49 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-PD1 light chain CDRL2 of the array number 50 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-PD1 light chain CDRL3 of the array number 51 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-PD1 heavy chain CDRH1 of the array number 52 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-PD1 heavy chain CDRH2 of the array number 53 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-PD1 heavy chain CDRH3 of the array number 54 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-CTLA-4 light chain CDRL1 of the array number 55 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-CTLA-4 light chain CDRL2 of the array number 56 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-CTLA-4 light chain CDRL3 of the array number 57 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-CTLA-4 heavy chain CDRH1 of the array number 58 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-CTLA-4 heavy chain CDRH2 of the array number 59 - anti-PD-1 / CTRL-4 bispecific antibody Amino acid sequence of the anti-CTLA-4 heavy chain CDRH3 of the array number 60 - anti-PD-1 / CTRL-4 bispecific antibody

Claims

**Claim 1** A pharmaceutical product comprising an antibody-drug conjugate and a bispecific checkpoint inhibitor for combined administration, wherein the antibody-drug conjugate has a drug-linker represented by the following formula: 【Chemical 1】 An antibody-drug conjugate represented by the formula: wherein A represents the position of connection to the antibody and is conjugated to the antibody, preferably an anti-TROP2 or anti-HER2 antibody, via a thioether bond. A pharmaceutical product. **Claim 2** The pharmaceutical product according to claim 1, wherein the drug-linker is conjugated to an anti-TROP2 antibody. **Claim 3** The anti-TROP2 antibody comprises a heavy chain comprising CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and a light chain comprising CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, CDRL2 consisting of the amino acid sequence represented by SEQ ID NO: 7, and CDRL3 consisting of the amino acid sequence represented by SEQ ID NO:

8. The pharmaceutical product according to claim 2. **Claim 4** The anti-TROP2 antibody is an antibody comprising a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 9 and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO:

10. The pharmaceutical product according to claim 3. **Claim 5** The anti-TROP2 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 12 and a light chain consisting of the amino acid sequence represented by SEQ ID NO:

13. The pharmaceutical product according to claim 4. **Claim 6** The anti-TROP2 antibody is an antibody comprising a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 11 and a light chain consisting of the amino acid sequence represented by SEQ ID NO:

13. The pharmaceutical product according to claim 4. **Claim 7** The pharmaceutical product according to any one of claims 2 to 6, wherein the average number of conjugated drug-linkers per anti-TROP2 antibody molecule in the antibody-drug conjugate ranges from 3.5 to 4.

5. **Claim 8** The pharmaceutical product according to claim 7, wherein the anti-TROP2 antibody-drug conjugate is datopotamab deruxtecan (DS-1062). **Claim 9** The pharmaceutical product according to claim 1, wherein the drug-linker is conjugated to an anti-HER2 antibody. **Claim 10** The pharmaceutical product according to claim 9, wherein the anti-HER2 antibody comprises a heavy chain comprising CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 16, CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 17, and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 18, and a light chain comprising CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 19, CDRL2 consisting of the amino acid sequence consisting of amino acid residues 1 to 3 of SEQ ID NO: 20, and CDRL3 consisting of the amino acid sequence represented by SEQ ID NO:

21.

11. The pharmaceutical product according to claim 10, wherein the anti-HER2 antibody comprises a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22, and a light chain comprising a light chain variable region consisting of the amino acid sequence represented by SEQ ID NO:

23.

12. The pharmaceutical product according to claim 11, wherein the anti-HER2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 14 and a light chain consisting of the amino acid sequence represented by SEQ ID NO:

15.

13. The pharmaceutical product according to claim 11, wherein the anti-HER2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 24 and a light chain consisting of the amino acid sequence represented by SEQ ID NO:

15.

14. The pharmaceutical product according to any one of claims 9 to 13, wherein the average number of conjugated drug-linker units per anti-HER2 antibody molecule in the antibody-drug conjugate ranges from 7 to 8.

15. The pharmaceutical product according to claim 14, wherein the anti-HER2 antibody-drug conjugate is trastuzumab deruxtecan (DS-8201).

16. The pharmaceutical product according to any one of claims 1 to 15, wherein the bispecific checkpoint inhibitor is a bispecific binding protein comprising a first binding domain that specifically binds to PD-1 and a second binding domain that specifically binds to CTLA-4 or TIGIT.

17. The bispecific binding protein is a) A first binding domain that specifically binds to PD-1, comprising a heavy chain variable domain having a CDRH1 with the amino acid sequence of SEQ ID NO: 25, a CDRH2 with the amino acid sequence of SEQ ID NO: 26, and a CDRH3 with the amino acid sequence of SEQ ID NO: 27, and a light chain variable domain having a CDRL1 with the amino acid sequence of SEQ ID NO: 28, a CDRL2 with the amino acid sequence of SEQ ID NO: 29, and a CDRL3 with the amino acid sequence of SEQ ID NO: 30, the first binding domain; b) A second binding domain that specifically binds to TIGIT, comprising a heavy chain variable domain having a CDRH1 with the amino acid sequence of SEQ ID NO: 35, a CDRH2 with the amino acid sequence of SEQ ID NO: 36, and a CDRH3 with the amino acid sequence of SEQ ID NO: 37, and a light chain variable domain having a CDRL1 with the amino acid sequence of SEQ ID NO: 38, a CDRL2 with the amino acid sequence of SEQ ID NO: 39, and a CDRL3 with the amino acid sequence of SEQ ID NO: 40, the second binding domain, the pharmaceutical product according to claim 16.

18. The pharmaceutical product according to claim 17, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 31 and a light chain variable domain having the amino acid sequence of SEQ ID NO:

33.

19. The pharmaceutical product according to claim 17, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 31 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

33.

20. The pharmaceutical product according to any one of claims 17 to 19, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 32 and a light chain having the amino acid sequence of SEQ ID NO:

34.

21. The pharmaceutical product according to any one of claims 17 to 19, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 32 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

34.

22. The pharmaceutical product according to any one of claims 17 to 21, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having the amino acid sequence of SEQ ID NO:

43.

23. The pharmaceutical product according to any one of claims 17 to 21, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

43.

24. The pharmaceutical product according to any one of claims 17 to 23, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino acid sequence of SEQ ID NO: 42 and a light chain having the amino acid sequence of SEQ ID NO:

44.

25. The pharmaceutical product according to any one of claims 17 to 23, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 42 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

44.

26. The bispecific binding protein is a) a first binding domain that specifically binds to PD-1, the first binding domain comprising a heavy chain variable domain comprising CDRH1 having the amino acid sequence of SEQ ID NO: 52, CDRH2 having the amino acid sequence of SEQ ID NO: 53, and CDRH3 having the amino acid sequence of SEQ ID NO: 54, and a light chain variable domain comprising CDRL1 having the amino acid sequence of SEQ ID NO: 49, CDRL2 having the amino acid sequence of SEQ ID NO: 50, and CDRL3 having the amino acid sequence of SEQ ID NO: 51; and b) a second binding domain that specifically binds to CTLA-4, the second binding domain comprising a heavy chain variable domain comprising CDRH1 having the amino acid sequence of SEQ ID NO: 58, CDRH2 having the amino acid sequence of SEQ ID NO: 59, and CDRH3 having the amino acid sequence of SEQ ID NO: 60, and a light chain variable domain comprising CDRL1 having the amino acid sequence of SEQ ID NO: 55, CDRL2 having the amino acid sequence of SEQ ID NO: 56, and CDRL3 having the amino acid sequence of SEQ ID NO: 57; the pharmaceutical product according to claim 16.

27. The pharmaceutical product according to claim 26, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 46 and a light chain having the amino acid sequence of SEQ ID NO:

45.

28. The pharmaceutical product according to claim 26, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 46 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

45.

29. The pharmaceutical product according to any one of claims 26 to 28, wherein the second binding domain that specifically binds to CTLA-4 comprises a heavy chain having the amino sequence of SEQ ID NO: 48 and a light chain having the amino acid sequence of SEQ ID NO:

47.

30. The pharmaceutical product according to any one of claims 26 to 28, wherein the second binding domain that specifically binds to CTLA-4 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 48 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

47.

31. The pharmaceutical product according to claim 20 or 27, wherein the light chain constant region is a kappa chain.

32. The pharmaceutical product according to claim 24 or 29, wherein the light chain constant region is a lambda chain.

33. The pharmaceutical product according to any one of claims 16 to 32, wherein the binding protein is an antibody.

34. The pharmaceutical product according to claim 33, wherein the antibody is an IgG antibody.

35. The pharmaceutical product according to claim 34, wherein the antibody is an IgG1 antibody.

36. The pharmaceutical product according to claim 34 or 35, wherein the antibody is a human antibody or a humanized antibody.

37. The pharmaceutical product according to any one of claims 33 to 36, wherein the bispecific antibody is monovalent.

38. The pharmaceutical product according to any one of claims 16 to 37, wherein the bispecific binding protein is a DuoBody.

39. The pharmaceutical product according to any one of claims 16 to 38, wherein the bispecific binding protein comprises a mutated Fc region.

40. When the mutant Fc region is numbered by the EU index described in Kabat, 221K, 221Y, 225E, 225K, 225W, 228P, 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235E, 235F, 236E, 237L, 237M, 237P, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 240I, 240A, 240T, 240M, 241W, 241L, 241Y, 241E, 241R, 243W, 243L, 243Y, 243R, 243Q, 244H, 245A, 247L, 247V, 247G, 250E, 250Q, 251F, 252L, 252Y, 254S, 254T, 255L, 256E, 256F, 256M, 257C, 257M, 257N, 262I, 262A, 262T, 262E, 263I, 263A, 263T, 263M, 264L, 264I, 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265A, 265G, 265N, 265Q, 265Y, 265F, 265V, 265I, 265L, 265H, 265T, 266I, 266A, 266T, 266M, 267Q, 267L, 268E, 269H, 269Y, 269F, 269R, 270E, 280A, 284M, 292P, 292L, 296E, 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H, 296G, 297S, 297D, 297E, 298A, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 305I, 308F, 313F, 316D, 318A, 318S, 320A, 320S, 322A, 322S, 325Q, 325L, 3251, 325D, 325E, 325A, 325T, 325V, 325H, 326A, 326D, 326E, 326G, 326M, 326V, 327G, 327W, 327N, 327L, 328S, 328M, 328D, 328E, 328N, 328Q, 328F, 3281, 328V, 328T, 328H, 328A, 329F, 329H, 329Q, 330K,The pharmaceutical product according to claim 39, comprising at least one substitution selected from 330G, 330T, 330C, 330L, 330Y, 330V, 3301, 330F, 330R, 330H, 331G, 331A, 331L, 331M, 331F, 331W, 331K, 331Q, 331E, 331S, 331V, 3311, 331C, 331Y, 331H, 331R, 331N, 331D, 331T, 332D, 332S, 332W, 332F, 332E, 332N, 332Q, 332T, 332H, 332Y, 332A, 333A, 333D, 333G, 333Q, 333S, 333V, 334A, 334E, 334H, 334L, 334M, 334Q, 334V, 334Y, 339T, 370E, 370N, 378D, 392T, 396L, 416G, 419H, 421K, 428L, 428F, 433K, 433L, 434A, 434W, 434Y, 436H, 440Y and 443W.,

41. The pharmaceutical product according to claim 39, wherein the mutated Fc region comprises one or more amino acid substitutions at positions selected from 428 and 434 numbered according to the EU index described in Kabat.

42. The pharmaceutical product according to any one of claims 39 to 41, wherein the mutated Fc region comprises one or more amino acid substitutions selected from 428L, 428F, 434A, 434W, and 434Y.

43. The pharmaceutical product according to any one of claims 39 to 42, wherein the mutated Fc region comprises the L234F / L235E / P331S triple mutation (TM).

44. The pharmaceutical product according to any one of claims 16 to 43, wherein the bispecific binding protein comprises an unglycosylated Fc region.

45. The pharmaceutical product according to any one of claims 16 to 43, wherein the bispecific binding protein comprises a deglycosylated Fc region.

46. The pharmaceutical product according to any one of claims 16 to 43, wherein the bispecific binding protein comprises an Fc region having reduced fucosylation or no fucosylation.

47. The pharmaceutical product according to any one of claims 1 to 46, wherein the product is a combination preparation comprising the antibody-drug conjugate and the bispecific checkpoint inhibitor for separate simultaneous administration.

48. The pharmaceutical product according to any one of claims 1 to 46, wherein the product is a combination preparation comprising the antibody-drug conjugate and the bispecific checkpoint inhibitor for sequential or separate simultaneous administration.

49. The pharmaceutical product according to any one of claims 1 to 48, wherein the product is for treating cancer.

50. The pharmaceutical product according to claim 49, wherein the cancer is at least one selected from the group consisting of 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 cancer, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumor, gastrointestinal 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, multiple myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer, and renal cell carcinoma.

51. The pharmaceutical product according to claim 50, wherein the cancer is breast cancer.

52. The pharmaceutical product according to claim 51, wherein the breast cancer is HER2-positive breast cancer.

53. The pharmaceutical product according to claim 51, wherein the breast cancer is HER2-low breast cancer.

54. The pharmaceutical product according to claim 51, wherein the breast cancer is triple-negative breast cancer.

55. The pharmaceutical product according to claim 51, wherein the breast cancer is hormone receptor (HR)-positive and HER2-negative breast cancer.

56. The pharmaceutical product according to claim 50, wherein the cancer is lung cancer.

57. The pharmaceutical product according to claim 56, wherein the lung cancer is non-small cell lung cancer.

58. The pharmaceutical product according to claim 57, wherein the non-small cell lung cancer has actionable genomic alterations.

59. The pharmaceutical product according to claim 57, wherein the non-small cell lung cancer has no actionable genomic alterations.

60. The pharmaceutical product according to claim 50, wherein the cancer is colorectal cancer.

61. The pharmaceutical product according to claim 50, wherein the cancer is gastric cancer.

62. The pharmaceutical product according to claim 50, wherein the cancer is pancreatic cancer.

63. The pharmaceutical product according to claim 50, wherein the cancer is ovarian cancer.

64. The pharmaceutical product according to claim 50, wherein the cancer is prostate cancer.

65. The pharmaceutical product according to claim 50, wherein the cancer is renal cancer.

66. The pharmaceutical product according to claim 50, wherein the cancer is bladder cancer.

67. The pharmaceutical product according to claim 50, wherein the cancer is endometrial cancer.

68. The pharmaceutical product according to claim 50, wherein the cancer is biliary tract cancer.

69. The pharmaceutical product according to any one of claims 1 to 48 for use in the treatment of cancer.

70. The pharmaceutical product for use according to claim 69, wherein the cancer is as described in any one of claims 50 to 68.

71. The pharmaceutical product according to any one of claims 1 to 48, further comprising carboplatin for administration in combination with the antibody-drug conjugate and the bispecific checkpoint inhibitor.

72. The pharmaceutical product according to any one of claims 1 to 48, further comprising fluoropyrimidine for administration in combination with the antibody-drug conjugate and the bispecific checkpoint inhibitor.

73. Use of an antibody-drug conjugate in the manufacture of a medicament for use in combination with a bispecific checkpoint inhibitor, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as described in any one of claims 1 to 46 for treating cancer.

74. The use according to claim 73, wherein the medicament is for use in combination with the bispecific checkpoint inhibitor by continuous administration.

75. The use according to claim 73, wherein the medicament is for use in combination with the bispecific checkpoint inhibitor by separate simultaneous administration.

76. Use of a bispecific checkpoint inhibitor in the manufacture of a medicament for use in combination with an antibody-drug conjugate, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as defined in any one of claims 1 to 46 for treating cancer.

77. The use according to claim 76, wherein the medicament is for use in combination with the antibody-drug conjugate by continuous administration.

78. The use according to claim 76, wherein the medicament is for use in combination with the antibody-drug conjugate by separate simultaneous administration.

79. The use according to any one of claims 73 to 78, wherein the cancer is as defined in any one of claims 50 to 68.

80. An antibody-drug conjugate for use in combination with a bispecific checkpoint inhibitor in the treatment of cancer, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as defined in any one of claims 1 to 46.

81. The antibody-drug conjugate for use according to claim 80, wherein the cancer is as defined in any one of claims 50 to 68.

82. The antibody-drug conjugate for use according to claim 80 or 81, wherein the use comprises continuously administering the antibody-drug conjugate and the bispecific checkpoint inhibitor.

83. The antibody-drug conjugate for use according to claim 80 or 81, wherein the use comprises administering the antibody-drug conjugate and the bispecific checkpoint inhibitor separately or simultaneously.

84. An antibody-drug conjugate for use in the treatment of cancer in a subject, wherein the treatment comprises i) continuous or separate simultaneous administration of the antibody-drug conjugate and ii) the bispecific checkpoint inhibitor to the subject, and the antibody-drug conjugate and the bispecific checkpoint inhibitor are as described in any one of claims 1 to 46.

85. A bispecific checkpoint inhibitor for use in combination with an antibody-drug conjugate in the treatment of cancer, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as described in any one of claims 1 to 46.

86. The bispecific checkpoint inhibitor for use according to claim 85, wherein the cancer is as described in any one of claims 50 to 68.

87. The bispecific checkpoint inhibitor for use according to claim 85 or 86, wherein the use comprises continuously administering the antibody-drug conjugate and the bispecific checkpoint inhibitor.

88. The bispecific checkpoint inhibitor for use according to claim 85 or 86, wherein the use comprises administering the antibody-drug conjugate and the bispecific checkpoint inhibitor separately or simultaneously.

89. A bispecific checkpoint inhibitor for use in the treatment of cancer in a subject, wherein the treatment comprises i) continuous or separate simultaneous administration of the bispecific checkpoint inhibitor and ii) the antibody-drug conjugate to the subject, and the bispecific checkpoint inhibitor and the antibody-drug conjugate are as described in any one of claims 1 to 46.

90. A method of treating cancer, comprising administering in combination to a subject in need thereof an antibody-drug conjugate and a bispecific checkpoint inhibitor as described in any one of claims 1 to 46.

91. The method according to claim 90, wherein the cancer is as described in any one of claims 50 to 68.

92. The method according to claim 90 or 91, wherein the method comprises continuously administering the antibody-drug conjugate and the bispecific checkpoint inhibitor.

93. The method according to claim 90 or 91, comprising administering the antibody-drug conjugate and the bispecific checkpoint inhibitor separately and simultaneously.

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