Combination of antibody-drug conjugates and bispecific checkpoint inhibitors

Abstract

A pharmaceutical product for the administration of an antibody-drug conjugate in combination with a bispecific checkpoint inhibitor is provided. This antibody-drug conjugate is an antibody-drug conjugate in which a drug-linker represented by the following formula (wherein A represents a connection position to an antibody) is conjugated to an antibody, preferably an anti-TROP2 or 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 to a subject in combination (Formula I). # imgabs0 #

Description

Technical Field

[0001] The present disclosure relates to a pharmaceutical product for administering a specific-drug conjugate in combination with a bispecific check point inhibitor, and to therapeutic uses and methods in which 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 check point inhibitor combination are administered to a subject. Background Art

[0002] Immune checkpoint inhibitors are agents that inhibit the immunosuppressive system and activate anti-tumor 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 immune receptor present on certain T cells and natural killer cells (NK), is another target. Examples of known immune checkpoint inhibitors include the anti-PD-1 antibodies nivolumab (WO 2006 / 121168) and pembrolizumab (WO 2008 / 156712); the anti-PD-L1 antibodies atezolizumab (WO 2010 / 077634), durvalumab (WO 2011 / 066389) and avelumab (WO 2013 / 079174); the anti-CTLA-4 antibodies ipilimumab (WO 2001 / 014424) and tremelimumab (WO 2000 / 037504).

[0003] Bispecific binding proteins that specifically bind to two immune checkpoint targets are under development. Examples include the PD-1 / CTLA-4 bispecific antibodies AK104 (cadonilimab) and MEDI5752 (U.S. Patent 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 within cancer cells, leading to cancer cell death (Ducry, L., et al., Bioconjugate Chem. (2010) 21, 5-13; Alley, SC et al., Current Opinion in Chemical Biology (2010) 14, 529-537; Damle N.K. Expert Opin. Biol. Ther. (2004) 4, 1445-1452; Senter PD 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 consists of an antibody targeting HER2 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). DS-8201) has shown significant clinical efficacy in solid tumors expressing HER2, including breast cancer, gastric cancer, colorectal cancer, and non-small cell lung cancer. It is worth noting that among the above indications, DS-8201 has shown promising activity in tumors with low HER2 expression.

[0006] Another such antibody-drug conjugate is datopotamab deruxtecan (DS-1062), which consists of an antibody targeting TROP2 and a derivative of exatecan. In particular, WO 2015 / 098099 and WO2020 / 240467 provide detailed descriptions of exemplary antibody-drug conjugates targeting TROP2, including datopotamab deruxtecan (DS-1062). Datopotamab has shown clinical efficacy in a variety of tumor types, including lung cancer and breast cancer.

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

[0008] However, further identification of combination partners for antibody-drug conjugates, including anti-HER2 antibody-drug conjugates such as DS-8201 and anti-TROP2 antibody-drug conjugates such as DS-1062, is needed to enhance their therapeutic potential.

[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 therapeutic responses, improve patient tolerability, reduce dose-dependent toxicities, and / or provide alternative treatments for cancers that have demonstrated resistance or refractory to previous cancer therapies. Summary of the invention

[0010] Antibody-drug conjugates (e.g., anti-TROP2 or anti-HER2 antibody-drug conjugates) comprising derivatives of topoisomerase I inhibitor exatecan as components used in the present disclosure have been shown to exhibit excellent anti-tumor effects in the treatment of certain cancers such as breast cancer when administered alone or in combination with checkpoint inhibitors. In addition, it has been shown that bispecific checkpoint inhibitors exhibit anti-tumor effects in the treatment of certain cancers. However, it is desirable to provide a drug and treatment that can obtain excellent anti-tumor effects in the treatment of cancer, such as enhanced efficacy, increased persistence of therapeutic response, and / or reduced dose-dependent toxicity.

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

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

[93] :

[0013] [1] A pharmaceutical product comprising an antibody-drug conjugate and a bispecific checkpoint inhibitor for combined administration, wherein the antibody-drug conjugate is an antibody-drug conjugate in which the drug-linker is represented by the following formula:

[0014]

[0015] Wherein A represents the connection position to the antibody, which is conjugated to the antibody via a thioether bond, preferably an anti-TROP2 or anti-HER2 antibody;

[0016] [2] The drug product according to [1], wherein the drug-linker is conjugated to an anti-TROP2 antibody;

[0017] [3] The pharmaceutical product according to [2], wherein the anti-TROP2 antibody comprises a heavy chain and

[0018] An antibody comprising a light chain, wherein the heavy chain comprises 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 the light chain comprises 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;

[0019] [4] The pharmaceutical product according to [3], wherein the anti-TROP2 antibody comprises a heavy chain and

[0020] The antibody comprises a heavy chain variable region consisting of an amino acid sequence represented by SEQ ID NO: 9, and a light chain variable region consisting of an amino acid sequence represented by SEQ ID NO: 10.

[0021] A light chain variable region composed of an amino acid sequence;

[0022] [5] The pharmaceutical product according to [4], wherein the anti-TROP2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 12 and a

[0023] An antibody having a light chain composed of the amino acid sequence shown;

[0024] [6] The pharmaceutical product according to [4], wherein the anti-TROP2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 11 and a

[0025] An antibody having a light chain composed of the amino acid sequence shown;

[0026] [7] The pharmaceutical product according to any one of [2] to [6], wherein the antibody-drug conjugate

[0027] The average unit of the drug-linker conjugated to each anti-TROP2 antibody molecule in the composition

[0028] The number is in the range of 3.5 to 4.5;

[0029] [8] The pharmaceutical product according to [7], wherein the anti-TROP2 antibody-drug conjugate is

[0030] Tecan (DS-1062);

[0031] [9] The drug product according to [1], wherein the drug-linker is conjugated to an anti-HER2 antibody

[0032] body;

[0033]

[10] The pharmaceutical product according to [9], wherein the anti-HER2 antibody comprises a heavy chain and a light chain.

[0034] The heavy chain comprises a CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 16, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 17

[0035] and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO:18, the 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;

[0036]

[11] The pharmaceutical product according to

[10] , wherein the anti-HER2 antibody comprises a heavy chain and

[0037] The antibody comprises a heavy chain variable region consisting of an amino acid sequence represented by SEQ ID NO: 22, and a light chain variable region consisting of an amino acid sequence represented by SEQ ID NO: 23.

[0038] A light chain variable region composed of an amino acid sequence;

[0039]

[12] The pharmaceutical product according to

[11] , wherein the anti-HER2 antibody comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 14 and a

[0040] An antibody having a light chain composed of the amino acid sequence shown;

[0041]

[13] The pharmaceutical product according to

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

[0042] An antibody having a light chain composed of the amino acid sequence shown;

[0043]

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

[13] , wherein the antibody-drug conjugate

[0044] The average number of units of the drug-linker conjugated to each anti-HER2 antibody molecule in the compound

[0045] The number of primaries is in the range of 7 to 8;

[0046]

[15] The pharmaceutical product according to

[14] , wherein the anti-HER2 antibody-drug conjugate is Dequ

[0047] Tocilizumab (DS-8201);

[0048]

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

[15] , wherein the bispecific assay

[0049] The inhibitor is a protein that contains a first binding domain that specifically binds to PD-1 and a specific binding domain.

[0050]

[17] A bispecific binding protein that binds to the second binding domain of CTLA-4 or TIGIT;

[17] The pharmaceutical product according to

[16] , wherein the bispecific binding protein comprises:

[0051] a) a first binding domain that specifically binds to PD-1, wherein the first binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDRH1 having an amino acid sequence of SEQ ID NO: 25, CDRH2 having an amino acid sequence of SEQ ID NO: 26, and CDRH3 having an amino acid sequence of SEQ ID NO: 27, and the light chain variable domain comprises CDRL1 having an amino acid sequence of SEQ ID NO: 28, CDRL2 having an amino acid sequence of SEQ ID NO: 29, and CDRL3 having an amino acid sequence of SEQ ID NO: 30; and

[0052] b) a second binding domain that specifically binds to TIGIT, wherein the second binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDRH1 having an amino acid sequence of SEQ ID NO: 35, CDRH2 having an amino acid sequence of SEQ ID NO: 36, and CDRH3 having an amino acid sequence of SEQ ID NO: 37, and the light chain variable domain comprises CDRL1 having an amino acid sequence of SEQ ID NO: 38, CDRL2 having an amino acid sequence of SEQ ID NO: 39, and CDRL3 having an amino acid sequence of SEQ ID NO: 40;

[0053]

[18] The pharmaceutical product according to

[17] , wherein the first

[0054] The binding domain 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;

[0055]

[19] The pharmaceutical product according to

[17] , wherein the first binding agent specifically binds to PD-1

[0056] The domain 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;

[0057]

[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 an amino acid sequence of SEQ ID NO: 32 and a light chain having an amino acid sequence of SEQ ID NO: 34;

[0058]

[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 heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:

[0059] A light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of 34;

[0060]

[22] The 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 of SEQ ID NO: 41 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 43.

[0061] chain variable domain;

[0062]

[23] The 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 heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:43.

[0063] The light chain variable domain of the column;

[0064]

[24] The pharmaceutical product according to any one of

[17] to

[23] , wherein the second binding domain that specifically binds to TIGIT comprises an amino acid sequence having SEQ ID NO: 42

[0065] and a light chain having the amino acid sequence of SEQ ID NO: 44;

[0066]

[25] The 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 heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 43.

[0067] NO:44 has an amino acid sequence that is at least 90% identical to the light chain;

[26] The pharmaceutical product according to

[16] , wherein the bispecific binding protein comprises:

[0068] a) a first binding domain that specifically binds to PD-1, wherein the first binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDRH1 having an amino acid sequence of SEQ ID NO: 52, CDRH2 having an amino acid sequence of SEQ ID NO: 53, and CDRH3 having an amino acid sequence of SEQ ID NO: 54, and the light chain variable domain comprises CDRL1 having an amino acid sequence of SEQ ID NO: 49, CDRL2 having an amino acid sequence of SEQ ID NO: 50, and CDRL3 having an amino acid sequence of SEQ ID NO: 51; and

[0069] b) a second binding domain that specifically binds to CTLA-4, wherein the second binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprising CDRH1 having an amino acid sequence of SEQ ID NO:58, CDRH2 having an amino acid sequence of SEQ ID NO:59, and CDRH3 having an amino acid sequence of SEQ ID NO:60, and the light chain variable domain comprising CDRL1 having an amino acid sequence of SEQ ID NO:55, CDRL2 having an amino acid sequence of SEQ ID NO:56, and CDRL3 having an amino acid sequence of SEQ ID NO:

[0070] CDRL3 with an amino acid sequence of 57;

[0071]

[27] The pharmaceutical product according to

[26] , wherein the first

[0072] The binding domain 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;

[0073]

[28] The pharmaceutical product according to

[26] , wherein the first binding agent specifically binds to PD-1

[0074] The structural domain 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 heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:45.

[0075] light chains with amino acid sequences that are at least 90% identical to those of

[0076]

[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 an amino acid sequence of SEQ ID NO: 48 and a light chain having an amino acid sequence of SEQ ID NO: 47;

[0077]

[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;

[0078]

[31] The pharmaceutical product according to

[20] or

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

[0079]

[32] The pharmaceutical product according to

[24] or

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

[0080]

[33] The pharmaceutical product according to any one of

[16] to

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

[0081]

[34] The pharmaceutical product according to

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

[0082]

[35] The pharmaceutical product according to

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

[0083]

[36] The pharmaceutical product according to

[34] or

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

[37] The pharmaceutical product according to any one of

[33] to

[36] , wherein the bispecific antibody is monovalent;

[0084]

[38] The pharmaceutical product according to any one of

[16] to

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

[0085]

[39] The pharmaceutical product according to any one of

[16] to

[38] , wherein the bispecific binding protein comprises a variant Fc region;

[0086]

[40] The pharmaceutical product of

[39] , wherein the variant Fc region comprises at least one substitution selected from the group consisting of 221K, 221Y, 225E, 225K, 225W, 228P, 234D, 234E, 234N,

[0087] 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235A,

[0088] 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235E, 235F, 236E, 237L, 237M, 237P, 239D, 239E, 239N, 239Q, 239F, 239T,

[0089] 239H, 239Y, 240I, 240A, 240T, 240M, 241W, 241L, 241Y, 241E, 241R, 243W , 243L243Y, 243R, 243Q, 244H, 245A, 247L, 247V, 247G, 250E, 250Q, 251F,

[0090] <h2 style=";text-align:left;direction:ltr">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<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0091] <h2 style=";text-align:left;direction:ltr"> 265Y, 265F, 265V, 265I, 265L, 265H, 265T, 266I<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0092] <h2 style=";text-align:left;direction:ltr"> 266A, 266T, 266M, 267Q, 267L, 268E, 269H, 269Y<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0093] <h2 style=";text-align:left;direction:ltr"> 269F, 269R, 270E, 280A, 284M, 292P, 292L, 296E<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0094] <h2 style=";text-align:left;direction:ltr"> 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0095] <h2 style=";text-align:left;direction:ltr"> 296G, 297S, 297D, 297E, 298A, 298H, 298I, 298T<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0096] <h2 style=";text-align:left;direction:ltr"> 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0097] <h2 style=";text-align:left;direction:ltr"> 299E, 305I, 308F, 313F, 316D, 318A, 318S, 320A<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0098] <h2 style=";text-align:left;direction:ltr"> 320S, 322A, 322S, 325Q, 325L, 3251, 325D, 325E<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0099] <h2 style=";text-align:left;direction:ltr"> 325A, 325T, 325V, 325H, 326A, 326D, 326E, 326G<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0100] <h2 style=";text-align:left;direction:ltr"> 326M、326V、327G、327W、327N、327L、328S、328M、<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0101] <h2 style=";text-align:left;direction:ltr"> 328D、328E、328N、328Q、328F、3281、328V、328T、<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0102] <h2 style=";text-align:left;direction:ltr"> 328H, 328A, 329F, 329H, 329Q, 330K, 330G, 330T<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0103] 330C, 330L, 330Y, 330V, 3301, 330F, 330R, 330H,

[0104] 331G, 331A, 331L, 331M, 331F, 331W, 331K, 331Q,

[0105] 331E, 331S, 331V, 3311, 331C, 331Y, 331H, 331R,

[0106] 331N, 331D, 331T, 332D, 332S, 332W, 332F, 332E,

[0107] 332N, 332Q, 332T, 332H, 332Y, 332A, 333A, 333D,

[0108] 333G, 333Q, 333S, 333V, 334A, 334E, 334H, 334L,

[0109] 334M, 334Q, 334V, 334Y, 339T, 370E, 370N, 378D,

[0110] 392T, 396L, 416G, 419H, 421K, 428L, 428F, 433K,

[0111] 433L, 434A, 434W, 434Y, 436H, 440Y and 443W;

[0112]

[41] The pharmaceutical product of

[39] , wherein the variant Fc region comprises one or more amino acid substitutions at positions selected from 428 and 434 as numbered by the EU index as set forth in Kabat;

[0113]

[42] The pharmaceutical product according to any one of

[39] to

[41] , wherein the variant Fc region comprises one or more amino acid substitutions selected from 428L, 428F, 434A, 434W and 434Y;

[0114]

[43] The pharmaceutical product according to any one of

[39] to

[42] , wherein the variant Fc region comprises a L234F / L235E / P331S triple mutation (TM);

[0115]

[44] The pharmaceutical product according to any one of

[16] to

[43] , wherein the bispecific binding protein comprises an aglycosylated Fc region;

[0116]

[45] The pharmaceutical product according to any one of

[16] to

[43] , wherein the bispecific binding protein comprises a deglycosylated Fc region;

[0117]

[46] The pharmaceutical product according to any one of

[16] to

[43] , wherein the bispecific binding protein comprises an Fc region with reduced fucosylation or afucosylation;

[0118]

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

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

[0119]

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

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

[0120]

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

[48] , wherein the product is used to treat cancer;

[0121]

[50] The pharmaceutical product of

[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 carcinoma, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumors, gastrointestinal stromal tumors, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, uterine corpus cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasma cell neoplasm, myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer and renal cell carcinoma;

[0122]

[51] The pharmaceutical product according to

[50] , wherein the cancer is breast cancer;

[0123]

[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-expressing breast cancer;

[0124]

[54] The pharmaceutical product according to

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

[0125]

[55] The pharmaceutical product according to

[51] , wherein the breast cancer is a hormone receptor (HR)

[0126] positive, HER2-negative breast cancer;

[0127]

[56] The pharmaceutical product according to

[50] , wherein the cancer is lung cancer;

[0128]

[57] The pharmaceutical product according to

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

[0129]

[58] The pharmaceutical product according to

[57] , wherein the non-small cell lung cancer is a non-small cell lung cancer with a targetable genomic alteration;

[0130]

[59] The pharmaceutical product according to

[57] , wherein the non-small cell lung cancer is a non-small cell lung cancer without targetable genomic alterations;

[0131]

[60] The pharmaceutical product according to

[50] , wherein the cancer is colorectal cancer;

[0132]

[61] The pharmaceutical product according to

[50] , wherein the cancer is gastric cancer;

[0133]

[62] The pharmaceutical product according to

[50] , wherein the cancer is pancreatic cancer;

[0134]

[63] The pharmaceutical product according to

[50] , wherein the cancer is ovarian cancer;

[0135]

[64] The pharmaceutical product according to

[50] , wherein the cancer is prostate cancer;

[0136]

[65] The pharmaceutical product according to

[50] , wherein the cancer is renal cancer;

[0137]

[66] The pharmaceutical product according to

[50] , wherein the cancer is bladder cancer;

[0138]

[67] The pharmaceutical product according to

[50] , wherein the cancer is endometrial cancer;

[0139]

[68] The pharmaceutical product according to

[50] , wherein the cancer is biliary tract cancer;

[0140]

[69] A pharmaceutical product according to any one of [1] to

[48] , for use in treating cancer;

[0141]

[70] The pharmaceutical product for use according to

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

[50] to

[68] ;

[0142]

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

[48] , further comprising carboplatin for administration in combination with the antibody-drug conjugate and the bispecific checkpoint inhibitor;

[0143]

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

[48] , further comprising a fluoropyrimidine for administration in combination with the antibody-drug conjugate and the bispecific checkpoint inhibitor;

[0144]

[73] Use of an antibody-drug conjugate for the preparation of a medicament for treating cancer in combination with a bispecific checkpoint inhibitor, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as defined in any one of [1] to

[46] ;

[0145]

[74] The use according to

[73] , wherein the drug is used in combination with the bispecific checkpoint inhibitor by sequential administration;

[0146]

[75] The use according to

[73] , wherein the drug is used in combination with the bispecific checkpoint inhibitor by separate simultaneous administration;

[0147]

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

[46] ;

[0148]

[77] The use according to

[76] , wherein the drug is used in combination with the antibody-drug conjugate by sequential administration;

[0149]

[78] The use according to

[76] , wherein the drug is used in combination with the antibody-drug conjugate by separate simultaneous administration;

[0150]

[79] The use according to any one of

[73] to

[78] , wherein the cancer is as described in

[50] to

[68] .

[0151] as defined in any of the following;

[0152]

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

[46] ;

[0153]

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

[80] , wherein the cancer is such as [50 to

[0154]

[68] as defined in any one of the following:

[0155]

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

[80] or

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

[0156]

[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 and simultaneously;

[0157]

[84] An antibody-drug conjugate for use in treating cancer in a subject, wherein the treatment comprises sequentially or separately administering to the subject i) the antibody-drug conjugate and ii) a bispecific checkpoint inhibitor, wherein the antibody-drug conjugate and the bispecific checkpoint inhibitor are as defined in any one of [1] to

[46] ;

[0158]

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

[46] ;

[0159]

[86] A bispecific checkpoint inhibitor for use according to

[85] , wherein the cancer is

[0160] As defined in any of

[50] to

[68] ;

[0161]

[87] The bispecific checkpoint inhibitor for use according to

[85] or

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

[0162]

[88] The bispecific checkpoint inhibitor for use according to

[85] or

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

[0163]

[89] A bispecific checkpoint inhibitor for use in treating cancer in a subject, wherein the treatment comprises sequentially or separately administering to the subject i) the bispecific checkpoint inhibitor and ii) an antibody-drug conjugate, wherein the bispecific checkpoint inhibitor and the antibody-drug conjugate are as defined in any one of [1] to

[46] ;

[0164]

[90] A method for treating cancer, comprising administering to a subject in need thereof a combination of an antibody-drug conjugate according to any one of [1] to

[46] and a bispecific checkpoint inhibitor;

[0165]

[91] The method of

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

[50] to

[68] ;

[0166]

[92] The method according to

[90] or

[91] , wherein the method comprises sequentially administering the antibody-drug conjugate and the bispecific checkpoint inhibitor; and

[0167]

[93] The method of

[90] or

[91] , wherein the method comprises administering the antibody-drug conjugate and the bispecific checkpoint inhibitor separately and simultaneously.

[0168] [Beneficial Effects of the Present Disclosure]

[0169] The present disclosure provides a pharmaceutical product for combined administration comprising 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 in which the specific antibody-drug conjugate and the bispecific checkpoint inhibitor are administered in combination to a subject. Therefore, the present disclosure provides a drug and treatment that can obtain excellent anti-tumor effects in the treatment of cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0170] [Anti-TROP2 Antibody] :

[0171] Figure 1 The diagram shows the amino acid sequence (SEQ ID NO: 1) of the heavy chain of the anti-TROP2 antibody.

[0172] Figure 2 The diagram shows the amino acid sequence (SEQ ID NO: 2) of the light chain of the anti-TROP2 antibody.

[0173] Figure 3 The diagram shows the amino acid sequence of the heavy chain CDRH1 (SEQ ID NO: 3 [=amino acid residues 50 to 54 of SEQ ID NO: 1]).

[0174] Figure 4 The diagram shows the amino acid sequence of heavy chain CDRH2 (SEQ ID NO: 4 [=amino acid residues 69 to 85 of SEQ ID NO: 1]).

[0175] Figure 5 The figure shows the amino acid sequence of heavy chain CDRH3 (SEQ ID NO: 5 [=amino acid residues 118 to 129 of SEQ ID NO: 1]).

[0176] Figure 6 The diagram shows the amino acid sequence of the light chain CDRL1 (SEQ ID NO: 6 [=amino acid residues 44 to 54 of SEQ ID NO: 2]).

[0177] Figure 7 The diagram shows the amino acid sequence of the light chain CDRL2 (SEQ ID NO: 7 [=amino acid residues 70 to 76 of SEQ ID NO: 2]).

[0178] Figure 8 The diagram shows the amino acid sequence of light chain CDRL3 (SEQ ID NO: 8 [=amino acid residues 109 to 117 of SEQ ID NO: 2]).

[0179] Fig. 9 The diagram shows the amino acid sequence of the heavy chain variable region (SEQ ID NO: 9 [=amino acid residues 20 to 140 of SEQ ID NO: 1]).

[0180] Fig.10 The diagram shows the amino acid sequence of the light chain variable region (SEQ ID NO: 10 [=amino acid residues 21 to 129 of SEQ ID NO: 2]).

[0181] Fig.11 It is a diagram showing the amino acid sequence of the heavy chain (SEQ ID NO: 11 [=amino acid residues 20 to 469 of SEQ ID NO: 1]).

[0182] [Anti-HER2 Antibodies] :

[0183] Fig.12 The figure shows the amino acid sequence (SEQ ID NO: 14) of the heavy chain of the anti-HER2 antibody.

[0184] Fig.13 The figure shows the amino acid sequence (SEQ ID NO: 15) of the light chain of the anti-HER2 antibody.

[0185] Fig.14 The diagram shows the amino acid sequence of the heavy chain CDRH1 (SEQ ID NO: 16 [=amino acid residues 26 to 33 of SEQ ID NO: 14]).

[0186] Fig.15 The diagram shows the amino acid sequence of heavy chain CDRH2 (SEQ ID NO: 17 [=amino acid residues 51 to 58 of SEQ ID NO: 14]).

[0187] Fig.16 The diagram shows the amino acid sequence of heavy chain CDRH3 (SEQ ID NO: 18 [=amino acid residues 97 to 109 of SEQ ID NO: 14]).

[0188] Fig.17 The diagram shows the amino acid sequence of the light chain CDRL1 (SEQ ID NO: 19 [=amino acid residues 27 to 32 of SEQ ID NO: 15]).

[0189] Fig.18The diagram shows an amino acid sequence (SEQ ID NO: 20 [= amino acid residues 50 to 56 of SEQ ID NO: 15]) comprising the amino acid sequence of light chain CDRL2 (SAS).

[0190] Fig.19 The diagram shows the amino acid sequence of light chain CDRL3 (SEQ ID NO: 21 [=amino acid residues 89 to 97 of SEQ ID NO: 15]).

[0191] Fig. 20 The diagram shows the amino acid sequence of the heavy chain variable region (SEQ ID NO: 22 [=amino acid residues 1 to 120 of SEQ ID NO: 14]).

[0192] Fig.21 The diagram shows the amino acid sequence of the light chain variable region (SEQ ID NO: 23 [=amino acid residues 1 to 107 of SEQ ID NO: 15]).

[0193] Fig. 22 The diagram shows the amino acid sequence of the heavy chain (SEQ ID NO: 24 [=amino acid residues 1 to 449 of SEQ ID NO: 14]).

[0194] [Anti-PD-1 / TIGIT bispecific antibody] :

[0195] Fig.23 It is a diagram showing the amino acid sequence (SEQ ID NO: 25) of the anti-PD1 heavy chain CDRH1.

[0196] Fig.24 is a diagram showing the amino acid sequence of anti-PD1 heavy chain CDRH2 (SEQ ID NO: 26).

[0197] Fig.25 is a diagram showing the amino acid sequence of anti-PD1 heavy chain CDRH3 (SEQ ID NO: 27).

[0198] Fig.26 is a diagram showing the amino acid sequence (SEQ ID NO: 28) of the anti-PD1 light chain CDRL1.

[0199] Fig. 27 is a diagram showing the amino acid sequence of the anti-PD1 light chain CDRL2 (SEQ ID NO: 29).

[0200] Fig.28 It is a diagram showing the amino acid sequence (SEQ ID NO: 30) of the anti-PD1 light chain CDRL3.

[0201] Fig.29It is a diagram showing the amino acid sequence (SEQ ID NO: 31) of the anti-PD1 heavy chain variable region.

[0202] Fig.30 is a diagram showing the amino acid sequence of the anti-PD1 heavy chain (SEQ ID NO: 32).

[0203] Fig.31 It is a diagram showing the amino acid sequence (SEQ ID NO: 33) of the anti-PD1 light chain variable region.

[0204] Fig.32 is a diagram showing the amino acid sequence of the anti-PD1 light chain (SEQ ID NO: 34).

[0205] Fig.33 is a diagram showing the amino acid sequence of anti-TIGIT heavy chain CDRH1 (SEQ ID NO: 35).

[0206] Fig.34 is a diagram showing the amino acid sequence of anti-TIGIT heavy chain CDRH2 (SEQ ID NO: 36).

[0207] Fig.35 is a diagram showing the amino acid sequence of anti-TIGIT heavy chain CDRH3 (SEQ ID NO: 37).

[0208] Fig.36 is a diagram showing the amino acid sequence (SEQ ID NO: 38) of the anti-TIGIT light chain CDRL1.

[0209] Fig.37 is a diagram showing the amino acid sequence (SEQ ID NO: 39) of the anti-TIGIT light chain CDRL2.

[0210] Fig.38 is a diagram showing the amino acid sequence of the anti-TIGIT light chain CDRL3 (SEQ ID NO: 40).

[0211] Fig.39 is a diagram showing the amino acid sequence of the anti-TIGIT heavy chain variable region (SEQ ID NO: 41).

[0212] Fig.40 is a diagram showing the amino acid sequence of the anti-TIGIT heavy chain (SEQ ID NO: 42).

[0213] Fig.41 is a diagram showing the amino acid sequence (SEQ ID NO: 43) of the anti-TIGIT light chain variable region.

[0214] Fig.42is a diagram showing the amino acid sequence of the anti-TIGIT light chain (SEQ ID NO: 44).

[0215] [Anti-PD-1 / CTLA-4 bispecific antibody]:

[0216] Fig.43 is a diagram showing the amino acid sequence of the anti-PD1 light chain (SEQ ID NO: 45).

[0217] Fig.44 is a diagram showing the amino acid sequence of the anti-PD1 heavy chain (SEQ ID NO: 46).

[0218] Fig.45 is a diagram showing the amino acid sequence of the anti-CTLA-4 light chain (SEQ ID NO: 47).

[0219] Fig.46 is a diagram showing the amino acid sequence of the anti-CTLA-4 heavy chain (SEQ ID NO: 48).

[0220] Fig.47 It is a diagram showing the amino acid sequence (SEQ ID NO: 49) of the anti-PD1 light chain CDRL1.

[0221] Fig.48 is a diagram showing the amino acid sequence (SEQ ID NO: 50) of the anti-PD1 light chain CDRL2.

[0222] Fig.49 is a diagram showing the amino acid sequence (SEQ ID NO: 51) of the anti-PD1 light chain CDRL3.

[0223] Fig.50 is a diagram showing the amino acid sequence of anti-PD1 heavy chain CDRH1 (SEQ ID NO: 52).

[0224] Fig.51 is a diagram showing the amino acid sequence of anti-PD1 heavy chain CDRH2 (SEQ ID NO: 53).

[0225] Fig.52 is a diagram showing the amino acid sequence of anti-PD1 heavy chain CDRH3 (SEQ ID NO: 54).

[0226] Fig.53 is a diagram showing the amino acid sequence of the anti-CTLA-4 light chain CDRL1 (SEQ ID NO: 55).

[0227] Fig.54 is a diagram showing the amino acid sequence of the anti-CTLA-4 light chain CDRL2 (SEQ ID NO: 56).

[0228] Fig.55 is a diagram showing the amino acid sequence of the anti-CTLA-4 light chain CDRL3 (SEQ ID NO: 57).

[0229] Fig.56 is a diagram showing the amino acid sequence of anti-CTLA-4 heavy chain CDRH1 (SEQ ID NO: 58).

[0230] Fig.57 is a diagram showing the amino acid sequence of anti-CTLA-4 heavy chain CDRH2 (SEQ ID NO: 59).

[0231] Fig.58 is a diagram showing the amino acid sequence of anti-CTLA-4 heavy chain CDRH3 (SEQ ID NO: 60).

[0232] [experiment] :

[0233] Fig.59 Graphs showing (A) mean change in tumor volume over time ± SEM (n = 10 mice / treatment group), and (B) change in tumor volume over time for individual mice (CR = complete remission) in BALB / c mice bearing EMT6 hHER2 tumors treated with 10 mg / kg DS-8201 alone or in combination with 10 mg / kg anti-PD-L1, 10 mg / kg anti-PD-L1 + 10 mg / kg anti-CTLA-4, and 10 mg / kg anti-PD-1 / TIGIT DuetMab.

[0234] Fig.60 Presented are graphs showing tumor growth rate in BALB / c mice bearing EMT6 hHER2 tumors treated with 10 mg / kg DS-8201 alone or in combination with 10 mg / kg anti-PD-L1, 10 mg / kg anti-PD-L1 + 10 mg / kg anti-CTLA-4, and 10 mg / kg anti-PD-1 / TIGIT DuetMab (n=10 mice / treatment group).

[0235] Fig.61 Represents a graph reporting pharmacodynamic changes assessed via flow cytometry 8 days after the start of treatment in BALB / c mice bearing EMT6hHER2 tumors treated with 10 mg / kg DS-8201 alone or in combination with 10 mg / kg anti-PD-1 / TIGIT DuetMab.

[0236] Fig.62Represents a graph reporting changes in CTLA-4 expression on tumor CD8+ T cells assessed via flow cytometry 10 days after the start of treatment in EMT6 hHER2 bearing BALB / c mice treated with 10 mg / kg DS-8201.

[0237] Fig.63 Presented is a graph showing tumor growth rate in CD34+ humanized mice bearing Caki-1 tumors treated with a single dose of DS-8201 at 10 mg / kg IV alone, MEDI5752 at 10 mg / kg IP every two weeks alone, or a combination of DS-8201 and MEDI5752 each administered at 10 mg / kg.

[0238] Fig.64 Shown is a graph reporting changes in peripheral blood populations of CD8+ and CD4+ T cells assessed via flow cytometry 22 days after initiation of treatment with DS-8201, MEDI5752, or the combination of DS-8201 and MEDI5752. DETAILED DESCRIPTION

[0239] In order to more readily understand the present disclosure, certain terms are first defined. Additional definitions are set forth throughout the detailed description.

[0240] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure relates. For example, Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd edition, 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd edition, 1999, Academic Press; and Oxford Dictionary Of Biochemistry And Molecular Biology, Revised Edition, 2000, Oxford University Press provide a general dictionary of many of the terms used in the present disclosure for the skilled person.

[0241] Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0242] Units, prefixes and symbols are denoted in their generally accepted form in the International System of Units (SI). Numerical ranges are inclusive of the values ​​defining the range.

[0243] It should be understood that wherever herein aspects are described with the language "comprising," other similar aspects described with "consisting of" and / or "consisting essentially of" are also provided.

[0244] The terms "inhibit" and "inhibition" can refer to a decrease in biological activity by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. Cell proliferation can be determined using art-recognized techniques that measure the rate of cell division, and / or the fraction of cells within a cell population that undergo cell division, and / or the rate of cell loss in a cell population due to terminal differentiation or cell death (e.g., thymidine incorporation).

[0245] The term "subject" refers to any animal (e.g., mammal), including but not limited to humans, non-human primates, rodents, etc., that will be the recipient of a particular treatment. In general, the terms "subject" and "patient" are used interchangeably herein with respect to human subjects.

[0246] The term "drug product" refers to a formulation in which the active ingredients are in a form that allows them to be biologically active, either as a composition containing all active ingredients (for simultaneous administration), or as a combination of individual compositions (combination preparations) each containing at least one but not all active ingredients (for sequential or simultaneous administration), and which does not contain additional components that are unacceptably toxic to the subject to whom the product will be administered. Such a product may be sterile. "Simultaneous 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 each administration. The time interval may be, for example, less than 24 hours, preferably less than 6 hours, more preferably less than 2 hours.

[0247] Terms such as "treating" or "treatment" or "to treat" or "alleviating" or "to alleviate" refer to (1) therapeutic measures that cure, slow, alleviate the symptoms of, and / or halt the development of a diagnosed pathological condition or disorder, and (2) prophylactic / preventative measures that prevent and / or slow the development of a targeted pathological condition or disorder. Thus, those in need of treatment include those already suffering from the disorder; those susceptible to suffering from the disorder; and those in need of prevention of the disorder. In certain aspects, a subject is successfully "treated" for cancer according to the methods of the present disclosure if the patient shows, for example, complete, partial, or transient remission of a certain type of cancer.

[0248] The terms "cancer", "tumor", "cancerous" and "malignant" refer to or describe the physiological condition in mammals that is typically 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 tumors, gastrointestinal stromal tumors, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, uterine corpus cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer, and renal cell carcinoma. Cancers include hematological malignancies such as acute myeloid leukemia, multiple myeloma, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, Burkitt's lymphoma, follicular lymphoma, and solid tumors such as breast cancer, lung cancer, neuroblastoma, and colon cancer.

[0249] As used herein, the term "cytotoxic drug" is broadly defined and refers to a substance that inhibits or prevents cell function and / or causes cell destruction (cell death) and / or exerts an anti-tumor / anti-proliferative effect. For example, a cytotoxic drug directly or indirectly prevents the development, maturation or spread of neoplastic tumor cells. The term also includes agents that cause only cytostatic effects rather than just cytotoxic effects. The term includes chemotherapeutic agents as described below.

[0250] The term "chemotherapeutic agent" is a subset of the term "cytotoxic drug" that includes natural or synthetic compounds.

[0251] According to the methods or uses of the present disclosure, the compounds of the present disclosure can be administered to patients to promote a positive therapeutic response to cancer. The term "positive therapeutic response" with respect to cancer treatment refers to an improvement in symptoms associated with the disease. For example, the improvement of the disease can be characterized as complete remission. The term "complete remission" refers to the absence of clinically detectable disease and any previous test results are normalized. Alternatively, the improvement of the disease can be classified as partial remission. "Positive therapeutic response" encompasses a reduction or inhibition of cancer progression and / or duration, a reduction or improvement in cancer severity, and / or an improvement in one or more symptoms thereof due to the administration of the compounds of the present disclosure. In specific aspects, such terms refer to one, two, or three or more results after the administration of the compounds of the present disclosure:

[0252] (1) Stabilization, reduction or elimination of cancer cell populations;

[0253] (2) stabilization or reduction of cancer growth;

[0254] (3) Damage to cancer formation;

[0255] (4) eradication, removal, or control of primary, regional, and / or metastatic cancer;

[0256] (5) Reduction in mortality;

[0257] (6) increase in disease-free, relapse-free, progression-free, and / or overall survival, duration, or rate;

[0258] (7) an increase in the response rate, durability of response, or number of patients who respond or are in remission;

[0259] (8) Reduction in hospitalization rates,

[0260] (9) Reduction in hospital stay,

[0261] (10) maintaining the size of the cancer and not increasing or increasing by less than 10%, preferably less than 5%, preferably less than 4%, preferably less than 2%, and

[0262] (11) An increase in the number of patients in remission.

[0263] (12) A reduction in the amount of adjuvant therapy (e.g., chemotherapy or hormonal therapy) otherwise required to treat cancer.

[0264] Clinical response can be assessed using screening techniques such as PET, magnetic resonance imaging (MRI) scans, x-ray radiographic imaging, computed tomography (CT) scans, flow cytometry or fluorescence activated cell sorter (FACS) analysis, histology, gross pathology, and blood chemistry, including but not limited to changes detectable by ELISA, RIA, chromatography, etc. In addition to these positive treatment responses, subjects undergoing treatment may experience the beneficial effects of improved symptoms associated with the disease.

[0265] As used herein, the term "antibody" refers to a protein that can recognize and specifically bind to an antigen. Ordinary or conventional mammalian antibodies include tetramers, which are usually composed of two pairs of identical polypeptide chains, each pair consisting of a "light" chain (usually having a molecular weight of about 25kDa) and a "heavy" chain (usually having a molecular weight of about 50kDa-70kDa). As used herein, the terms "heavy chain" and "light chain" refer to any immunoglobulin polypeptide having sufficient variable domain sequences to confer specificity to a target antigen. The amino terminal portion of each light chain and heavy chain usually includes a variable domain of about 100 to 110 or more amino acids, which is usually responsible for antigen recognition. As used herein, the terms "variable region" or "variable domain" are used interchangeably and are common in the art. The carboxyl terminal portion of each chain usually defines a constant domain responsible for effector function. Therefore, in naturally occurring antibodies, a full-length heavy chain immunoglobulin polypeptide includes a variable domain (V H ) and three constant domains (C H1 , C H2 and C H3 ) and C H1 and C H2 The hinge region between the VH domain and the C H3 The variable domain is at the carboxyl terminus, and the full-length light chain immunoglobulin polypeptide includes the variable domain (V L ) and constant domain (C L ), where V L The domain is at the amino terminus of the polypeptide and the C L The domain is at the carboxyl terminus. However, one skilled in the art will appreciate that the location of the domain in naturally occurring antibodies 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 and lambda light chains.

[0266] In full-length light and heavy chains, the variable and constant domains are usually connected by a "J" region of about 12 or more amino acids, wherein the heavy chain also includes a "D" region of about 10 or more amino acids. The variable region of each light / heavy chain pair usually forms an antigen binding site. The variable domains of naturally occurring antibodies usually present the same general structure of a relatively conservative framework region (FR) connected by three hypervariable regions (also referred to as complementary determining regions or CDRs). The CDRs from the two chains of each pair are usually aligned by the framework region, which can enable binding to specific epitopes. From the amino terminus to the carboxyl terminus, the variable domains of both light and heavy chains usually include the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4.

[0267] The term "antibody fragment" refers to a complete or full-length chain or portion of an antibody, typically the target binding or variable region. Examples of antibody fragments include, but are not limited to, Fab 、F ab' 、F (ab')2 and F v As used herein, the term "functional fragment" is generally synonymous with "antibody fragment" and, with respect to an antibody, may refer to a fragment such as an F v 、F ab 、F (ab')2 of antibody fragments.

[0268] Reference to the numbering of amino acid residues described herein is according to the EU numbering system (also described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed., Public Health Service, National Institutes of Health, Bethesda, MD. (1991)).

[0269] "Monoclonal" antibody or its antigen binding fragment refers to a homogeneous antibody or antigen binding fragment population that participates in highly specific binding of a single antigenic determinant or epitope. This is in contrast to polyclonal antibodies that typically include different antibodies for different antigenic determinants. The term "monoclonal" antibody or its antigen binding fragment includes complete and full-length monoclonal antibodies and antibody fragments (such as Fab, Fab', F(ab')2, Fv), single-chain (scFv) mutants, fusion proteins comprising antibody portions, and any other modified immunoglobulin molecules comprising antigen recognition sites. In addition, "monoclonal" antibody or its antigen binding fragment refers to such antibodies and their antigen binding fragments prepared in a manner including but not limited to by hybridomas, phage selection, recombinant expression, and transgenic animals.

[0270] As used herein, the term "human antibody" includes antibodies having variable regions and constant regions that substantially correspond to human germline immunoglobulin sequences. In some aspects, 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 aspects, human antibodies are produced in hybridoma cells. In other aspects, human antibodies are produced recombinantly. In some aspects, bispecific binding proteins are human antibodies or humanized antibodies.

[0271] As used herein, the term "antigen" or "target antigen" refers to a molecule or a portion of a molecule that can be recognized and bound by a binding protein of the present disclosure. A target antigen can be used in an animal to produce antibodies that can bind to an epitope of the antigen. A target antigen can have one or more epitopes.

[0272] As used herein, the term "epitope" refers to a region or structural element of an antigen that is recognized and bound by a binding protein of the present disclosure. More precisely, an epitope is a specific structure bound by the CDRs of a binding protein. An epitope may include a portion of a protein structural element, a carbohydrate, or even a lipid structure found in a membrane. When a binding protein preferentially recognizes its antigenic target in a complex mixture of proteins and / or macromolecules, it is said that it specifically binds to an antigen. The term "specific binding" refers to a binding protein that specifically binds to a molecule or fragment thereof (e.g., an antigen). A binding protein that specifically binds to a molecule or fragment thereof can be bound to other molecules using a lower affinity, as determined by, for example, an immunoassay, BIAcore, or other assays known in the art. In particular, an antibody or fragment that specifically binds to at least one molecule or fragment thereof can compete for molecules that are non-specifically bound. The present disclosure particularly encompasses antibodies with multiple specificities (e.g., antibodies that are specific to two or more discrete antigens). For example, a bispecific antibody can bind to two adjacent epitopes on a single target antigen, or can bind to two different antigens.

[0273] As used herein, the term "antigen binding site" refers to a site produced on the surface of a binding protein of the present disclosure, wherein an antigen or an epitope on an antigen is bound. The antigen binding site of a binding protein is typically described by reference to the loop structure produced by the complementary determining regions (CDRs) of the binding protein.

[0274] [Description of the implementation plan]

[0275] Hereinafter, a preferred mode for carrying out the present disclosure is described. The embodiment described below is given only as an example to illustrate a typical embodiment of the present disclosure and is not intended to limit the scope of the present disclosure.

[0276] 1. Antibody-drug conjugates

[0277] The antibody-drug conjugate used in the present disclosure is an antibody-drug conjugate in which a drug-linker represented by the following formula is conjugated to an antibody, preferably an anti-TROP2 or anti-HER2 antibody, via a thioether bond:

[0278]

[0279] Wherein A represents the connection position with the antibody.

[0280] 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 attached 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.

[0281] The drug-linker of the present disclosure includes 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, (also represented by chemical name: (1S,9S)-1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3',4':6,7]indolizino[1,2-b]quinoline-10,13(9H,15H)-dione) as a component, which is a topoisomerase I inhibitor. Exatecan is a camptothecin derivative with anti-tumor effect, represented by the following formula:

[0282]

[0283] The antibody-drug conjugates used in the present disclosure can also be represented by the following formula:

[0284]

[0285] Here, the drug-linker is conjugated to an antibody ("antibody-"), preferably an anti-TROP2 or anti-HER2 antibody, via a thioether bond. n has the same meaning as the so-called average number of conjugated drug molecules (DAR; drug to antibody ratio) and represents the average number of drug-linker units conjugated per antibody molecule.

[0286] After migrating into cancer cells, the antibody-drug conjugate used in the present disclosure is cleaved at the linker moiety to release a compound represented by the following formula:

[0287]

[0288] 2. Antibodies in Antibody-Drug Conjugates

[0289] The antibody in the antibody-drug conjugate used in the present disclosure is preferably an anti-TROP2 or anti-HER2 antibody, and can be derived from any species, preferably from humans, rats, mice or rabbits. In the case where the antibody is derived from a species other than humans, it is preferably chimerized or humanized using known techniques. The antibody can be a polyclonal antibody or a monoclonal antibody, and is preferably a monoclonal antibody.

[0290] The antibody in the antibody-drug conjugate used in the present disclosure is preferably an antibody having characteristics capable of targeting cancer cells, and is preferably an antibody having, for example, a property of recognizing cancer cells, a property of binding to cancer cells, a property of internalizing in cancer cells, and / or a cytocidal activity against cancer cells.

[0291] The binding activity of antibodies to cancer cells can be confirmed using flow cytometry. Antibody internalization into cancer cells can be confirmed by: (1) using a secondary antibody (fluorescently labeled) bound to a therapeutic antibody, visualizing the determination of the antibody incorporated into the cell under a fluorescence microscope (Cell Death and Differentiation (2008) 15, 751-761); (2) using a secondary antibody (fluorescently labeled) bound to a therapeutic antibody, measuring the determination of the fluorescence intensity incorporated into the cell (Molecular Biology of the Cell, Vol. 15, 5268-5282, December 2004); or (3) using a Mab-ZAP assay of an immunotoxin bound to a therapeutic antibody, wherein the toxin is released when incorporated into the cell to inhibit cell growth (BioTechniques 28: 162-165, January 2000). As an immunotoxin, a recombinant complex protein of the diphtheria toxin catalytic domain and protein G can be used.

[0292] The anti-tumor activity of the antibody can be confirmed in vitro by measuring the inhibitory activity on cell growth. For example, a cancer cell line overexpressing the target protein of the antibody is cultured, and different concentrations of the antibody are added to the culture system to measure the inhibitory activity on lesion formation, colony formation, and spheroid growth. The anti-tumor activity can be confirmed in vivo, for example, by administering the antibody to nude mice with transplanted cancer cell lines that highly express the target protein and measuring the changes in the cancer cells.

[0293] Since the compound conjugated in the antibody-drug conjugate exerts an anti-tumor effect, it is preferred but not necessary that the antibody itself should have an anti-tumor effect. In order to specifically and selectively exert the cytotoxic activity of the anti-tumor compound on cancer cells, it is important and also preferred that the antibody should have the property of internalization to migrate into cancer cells.

[0294] The antibodies (preferably anti-TROP2 or anti-HER2 antibodies) in the antibody-drug conjugates used in the present disclosure can be obtained by methods well known in the art. For example, the antibodies of the present disclosure can be obtained using methods commonly performed in the art, which include immunizing animals with antigenic polypeptides and collecting and purifying antibodies produced in vivo. The source of the antigen is not limited to humans, and animals can be immunized with antigens derived from non-human animals such as mice, rats, etc. In this case, the cross-reactivity of antibodies bound to the obtained heterologous antigens with human antigens can be tested to screen antibodies suitable for human diseases.

[0295] Alternatively, antibody-producing cells that produce antibodies against an antigen are fused with myeloma cells to establish hybridomas according to methods known in the art (e.g., Kohler and Milstein, Nature (1975) 256, pp. 495-497; and Kennet, R., ed., Monoclonal Antibodies, pp. 365-367, Plenum Press, NY (1980)) to obtain monoclonal antibodies.

[0296] Antigens can be obtained by genetically engineering host cells to produce genes encoding antigenic proteins. Specifically, a vector that allows the expression of the antigenic gene is prepared and transferred to a host cell to express the gene. The antigen thus expressed can be purified. Antibodies can also be obtained by immunizing animals with the above-mentioned genetically engineered antigen expressing cells or cell lines expressing the antigen.

[0297] The antibody (preferably anti-TROP2 or anti-HER2 antibody) in the antibody-drug conjugate used in the present disclosure is preferably a recombinant antibody obtained by artificial modification for the purpose of reducing heterologous antigenicity to humans, such as a chimeric antibody or a humanized antibody, or preferably an antibody having only a gene sequence of an antibody derived from humans, i.e., a human antibody. These antibodies can be prepared using known methods.

[0298] Examples of chimeric antibodies include antibodies in which the variable region and constant region are derived from different species, for example, antibodies in which the variable region of an antibody derived from mouse or rat is linked to the constant region of an antibody derived from human (Proc. Natl. Acad. Sci. USA, 81, 6851-6855, (1984)).

[0299] As humanized antibodies, there can be exemplified antibodies obtained by integrating only the complementarity determining region (CDR) of a heterologous antibody into an antibody of human origin (Nature (1986) 321, pp. 522-525), antibodies obtained by grafting a portion of the amino acid residues of the framework of a heterologous antibody as well as the CDR sequence of a heterologous antibody into a human antibody by a CDR grafting method (WO90 / 07861), and antibodies humanized using a gene conversion mutagenesis strategy (U.S. Patent No. 5,821,337).

[0300] As human antibodies, antibodies produced using human antibody-producing mice having human chromosome fragments containing genes for heavy and light chains of human antibodies can be exemplified (see Tomizuka, K. et al., Nature Genetics (1997) 16, pp. 133-143; Kuroiwa, Y. et al., Nucl. Acids Res. (1998) 26, pp. 3447-3448; Yoshida, H. et al., Animal Cell Technology: Basic and Applied Aspects vol. 10, pp. 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, pp. 722-727, etc.). As an alternative, antibodies obtained by phage display, which are selected from a human antibody library (see Wormstone, IM et al., Investigative Ophthalmology & Visual Science. (2002) 43 (7), pp. 2301-2308; Carmen, S. et al., Briefings in Functional Genomics and Proteomics (2002), 1 (2), pp. 189-203; Siriwardena, D. et al., Ophthalmology (2002) 109 (3), pp. 427-431, etc.) can be exemplified.

[0301] In the antibody in the antibody-drug conjugate used in the present invention, the modified variant of the antibody is also included. The modified variant refers to a variant obtained by chemically or biologically modifying the antibody according to the present disclosure. Examples of chemically modified variants include variants comprising the connection of a chemical part to an amino acid backbone, variants comprising the connection of a chemical part to an N-connected or O-connected sugar chain, etc. Examples of biologically modified variants include variants obtained by post-translational modification (such as N-connected or O-connected glycosylation, N-terminal or C-terminal processing, deamidation, isomerization of aspartic acid or oxidation of methionine), and variants in which methionine residues are added to the N-terminal by expression in a prokaryotic host cell. In addition, antibodies labeled to enable detection or separation of antibodies or antigens according to the present disclosure, such as enzyme-labeled antibodies, fluorescently labeled antibodies, and affinity-labeled antibodies are also included in the meaning of modified variants. Such modified variants of antibodies according to the present disclosure can be used to improve the stability and blood retention of antibodies, reduce their antigenicity, detect or separate antibodies or antigens, etc.

[0302] In addition, by adjusting the modification (glycosylation, defucosylation, etc.) of the polysaccharide connected to the antibody according to the present disclosure, the antibody-dependent cellular toxicity activity can be enhanced. As the technology of adjusting the modification of the polysaccharide of the antibody, those disclosed in WO99 / 54342, WO00 / 61739, WO02 / 31140, WO2007 / 133855, WO2013 / 120066, etc. are known. However, the technology is not limited thereto. In the antibody according to the present disclosure (preferably anti-TROP2 or anti-HER2 antibody), the modified antibody of the polysaccharide is also included.

[0303] It is known that the lysine residue at the carboxyl terminus of the heavy chain of an antibody produced in cultured mammalian cells is deleted (Journal of Chromatography A, 705: 129-134 (1995)), and it is also known that two amino acid residues (glycine and lysine) at the carboxyl terminus of the heavy chain of an antibody produced in cultured mammalian cells are deleted, and 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 function (activation of complement, antibody-dependent cellular toxicity, etc.) of the antibody. Therefore, in the antibodies according to the present disclosure (preferably anti-TROP2 or anti-HER2 antibodies), antibodies and functional fragments of antibodies subjected to such modifications are also included, and deletion variants in which one or two amino acids are deleted at the carboxyl terminus of the heavy chain, variants obtained by amidation of deletion variants (e.g., heavy chains in which the carboxyl terminal proline residue has been amidated), etc. are also included. The types of deletion variants 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 variants, as long as the antigen binding affinity and effector function are conserved. The two heavy chains constituting the antibody according to the present disclosure may be one type selected from the group consisting of the full-length heavy chain and the above-mentioned deletion variants, or may be a combination of two types selected therefrom. The ratio of the amount of each deletion variant may be affected by the type and culture conditions of the cultured mammalian cells producing the antibody according to the present disclosure; however, it may be preferred to exemplify an antibody in which one amino acid residue at the carboxyl terminus is deleted in both heavy chains of the antibody according to the present disclosure.

[0304] As the isotype of the antibody according to the present disclosure (preferably an anti-TROP2 or anti-HER2 antibody), for example, IgG (IgG1, IgG2, IgG3, IgG4) can be exemplified, and IgG1 or IgG2 can be preferably exemplified.

[0305] The antibodies suitable for producing the antibody-drug conjugates according to the present disclosure are not particularly limited to any specific antigen. However, anti-TROP2 antibodies and anti-HER2 antibodies can be preferably exemplified.

[0306] 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 an activity of internalization in a cell expressing TROP2 by binding to TROP2.

[0307] Examples of anti-TROP2 antibodies include hTINA1-H1L1 (WO 2015 / 098099), and datopotamab can be preferably exemplified.

[0308] In the present disclosure, the term "anti-HER2 antibody" refers to an antibody that specifically binds to HER2 (human epidermal growth factor receptor type 2; ErbB-2), and preferably has an activity of internalizing in cells expressing HER2 by binding to HER2.

[0309] Examples of the anti-HER2 antibody include trastuzumab (U.S. Patent No. 5,821,337) and pertuzumab (WO01 / 00245), and trastuzumab can be preferably exemplified.

[0310] 3. Preparation of Antibody-Drug Conjugates

[0311] The drug-linker intermediate used to prepare the antibody-drug conjugate according to the present disclosure is represented by the following formula:

[0312]

[0313] 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 prepared with reference to the descriptions in WO2014 / 057687, WO2015 / 098099, WO2015 / 115091, WO2015 / 155998, WO2019 / 044947, etc.

[0314] The antibody-drug conjugate used in the present disclosure can be prepared by reacting the above-mentioned drug-linker intermediate with an antibody (preferably an anti-TROP2 or anti-HER2 antibody) having a thiol group (also called a sulfhydryl group).

[0315] Antibodies with thiol groups (preferably anti-TROP2 or anti-HER2 antibodies) can be obtained by methods known in the art (Hermanson, GT, Bioconjugate Techniques, pp. 56-136, pp. 456-493, Academic Press (1996)). For example, by using 0.3 to 3 molar equivalents of a reducing agent such as tris(2-carboxyethyl)phosphine hydrochloride (TCEP) per interchain disulfide in an antibody and reacting with the antibody in a buffer solution containing a chelating agent such as ethylenediaminetetraacetic acid (EDTA), an antibody with thiol groups of interchain disulfides in the antibody partially or completely reduced can be obtained.

[0316] Furthermore, by using 2 to 20 molar equivalents of the drug-linker intermediate per antibody having a thiol group (preferably an anti-TROP2 or anti-HER2 antibody), an antibody-drug conjugate in which 2 to 8 drug molecules are conjugated per antibody molecule can be prepared.

[0317] The average number of conjugated drug molecules per antibody (preferably anti-TROP2 or anti-HER2 antibody) molecule of the prepared antibody-drug conjugate can be determined, for example, by a calculation method based on measuring the UV absorbance of the antibody-drug conjugate and its conjugation precursor at two wavelengths of 280 nm and 370 nm (UV method), or by a calculation method based on quantification by HPLC measurement of fragments obtained by treating the antibody-drug conjugate with a reducing agent (HPLC method).

[0318] The conjugation between the antibody (preferably anti-TROP2 or anti-HER2 antibody) and the 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 WO2014 / 057687, WO2015 / 098099, WO2015 / 115091, WO2015 / 155998, WO2017 / 002776, WO2018 / 212136, etc.

[0319] In the present invention, the term "anti-TROP2 antibody-drug conjugate" refers to an antibody-drug conjugate, such that the antibody in the antibody-drug conjugate according to the present invention is an anti-TROP2 antibody.

[0320] The anti-TROP2 antibody is preferably an antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises CDRH1 consisting of the amino acid sequence represented by SEQ ID NO:3 [= an 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 [= an 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 [= an amino acid sequence consisting of amino acid residues 118 to 129 of SEQ ID NO:1], and the light chain comprises CDRL1 consisting of the amino acid sequence represented by SEQ ID NO:6 [= an 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 [= an 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 [= an amino acid sequence consisting of amino acid residues 109 to 117 of SEQ ID NO:2],

[0321] More preferably, an antibody comprises a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO:9 [= an 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 [= an amino acid sequence consisting of amino acid residues 21 to 129 of SEQ ID NO:2], and

[0322] Even more preferred is an antibody 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 [= the amino acid sequence consisting of amino acid residues 21 to 234 of SEQ ID NO:2].

[0323] In the anti-TROP2 antibody-drug conjugate, the average number of drug-linker units conjugated per antibody molecule is preferably 2 to 8, more preferably 3 to 5, even more preferably 3.5 to 4.5, and even more preferably about 4.

[0324] Anti-TROP2 antibody-drug conjugates can be prepared with reference to the descriptions in WO 2015 / 098099 and WO 2017 / 002776.

[0325] In a preferred embodiment, the anti-TROP2 antibody-drug conjugate is delutec (DS-1062).

[0326] In the present disclosure, the term "anti-HER2 antibody-drug conjugate" refers to an antibody-drug conjugate, such that the antibody in the antibody-drug conjugate according to the present disclosure is an anti-HER2 antibody.

[0327] The anti-HER2 antibody is preferably an antibody comprising a heavy chain and a light chain, the heavy chain comprising CDRH1 consisting of an amino acid sequence consisting of amino acid residues 26 to 33 of SEQ ID NO: 14, CDRH2 consisting of an amino acid sequence consisting of amino acid residues 51 to 58 of SEQ ID NO: 14, and CDRH3 consisting of an amino acid sequence consisting of amino acid residues 97 to 109 of SEQ ID NO: 14, and the light chain comprising CDRL1 consisting of an amino acid sequence consisting of amino acid residues 27 to 32 of SEQ ID NO: 15, CDRL2 consisting of an amino acid sequence consisting of amino acid residues 50 to 52 of SEQ ID NO: 15, and CDRL3 consisting of an amino acid sequence consisting of amino acid residues 89 to 97 of SEQ ID NO: 15; and more preferably an antibody comprising a heavy chain and a light chain, the heavy chain comprising a heavy chain variable region consisting of an amino acid sequence consisting of amino acid residues 1 to 120 of SEQ ID NO: 14, and a light chain variable region consisting of an amino acid sequence consisting of amino acid residues 1 to 107 of SEQ ID NO: 15; and even more preferably comprising a heavy chain consisting of an amino acid sequence represented by SEQ ID NO: 14 and a light chain variable region consisting of an amino acid sequence consisting of SEQ ID NO: 14. The invention relates to an antibody comprising 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 an amino acid sequence consisting of all amino acid residues 1 to 214 of SEQ ID NO:15.

[0328] In the anti-HER2 antibody-drug conjugates, the average number of drug-linker units conjugated per antibody molecule 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.

[0329] The anti-HER2 antibody-drug conjugate used in the present disclosure can be prepared with reference to the description in WO2015 / 115091 and the like.

[0330] In a preferred embodiment, the anti-HER2 antibody-drug conjugate is trastuzumab (DS-8201).

[0331] 4. Dual-specific checkpoint inhibitors

[0332] In the present disclosure, the term "bispecific checkpoint inhibitor" refers to a bispecific binding protein. As used herein, a bispecific binding protein has binding specificity for different epitopes within at least two independent antigens (or targets) or the same antigen. An exemplary bispecific binding protein may bind to two different epitopes of a target, or may bind to two different targets. Other such binding proteins may combine a first target binding site with a second binding site of another target. In some aspects, the binding protein is a bispecific antibody.

[0333] In some aspects, bispecific antibodies provide additive and / or synergistic therapeutic effects derived from simultaneously targeting two antigens through administration of a single manufactured molecule.

[0334] In some aspects, antibody provided herein is a monovalent bispecific antibody (MBab). Monovalent bispecific antibody scaffold as described herein provides an excellent platform for generating bispecific antibodies, which meet all the benefits associated with bispecific antibodies, while reducing the above-mentioned potential treatment risks due to their monovalent properties. In addition, MBab provided herein is easy to express, stable and may have low immunogenicity. As used herein, the term "monovalent bispecific" (can be abbreviated as "MBab") refers to bispecific antibodies, wherein each arm can specifically bind to different target antigens, and for a given pair of different target antigens (A and B), MBab can be bound to a kind of in each. In some aspects, monovalent bispecific antibodies can specifically bind to two independent epitopes on two independent antigens (or targets) or the same antigen. Typically, monovalent bispecific antibodies include two different variable regions. In some aspects, the binding affinity for two independent antigens is roughly the same. In some aspects, the binding affinity for two independent antigens is different.

[0335] 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.

[0336] In a preferred embodiment, the bispecific binding protein comprises:

[0337] a) a first binding domain that specifically binds to PD-1, wherein the first binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises a CDRH1 having an amino acid sequence of SEQ ID NO: 25, a light chain variable domain having an amino acid sequence of SEQ ID NO:

[0338] The light chain variable domain comprises a CDRH1 having an amino acid sequence of SEQ ID NO: 28, a CDRL2 having an amino acid sequence of SEQ ID NO: 29, and a CDRL3 having an amino acid sequence of SEQ ID NO: 30; and

[0339] b) a second binding domain that specifically binds to TIGIT, wherein the second binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDRH1 having an amino acid sequence of SEQ ID NO: 35, CDRH2 having an amino acid sequence of SEQ ID NO: 36, and CDRH3 having an amino acid sequence of SEQ ID NO: 37, and the light chain variable domain comprises CDRL1 having an amino acid sequence of SEQ ID NO: 38, CDRL2 having an amino acid sequence of SEQ ID NO: 39.

[0340] and CDRL3 having the amino acid sequence of SEQ ID NO:40.

[0341] More preferably, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having an amino acid sequence of SEQ ID NO: 31 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 33. In some aspects, 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 aspects, 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.

[0342] More preferably, the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence of SEQ ID NO: 32 and a light chain having an amino acid sequence of SEQ ID NO: 34. In some aspects, 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 aspects, 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.

[0343] More preferably, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 43. In some aspects, 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 aspects, 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.

[0344] More preferably, the second binding domain that specifically binds to TIGIT comprises a heavy chain having an amino acid sequence of SEQ ID NO: 42 and a light chain having an amino acid sequence of SEQ ID NO: 44. In some aspects, 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 aspects, 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.

[0345] In a particularly preferred embodiment, the bispecific binding protein is an antibody comprising:

[0346] (a) a first binding domain that specifically binds to PD-1, the first binding domain comprising a heavy chain having an amino acid sequence of SEQ ID NO: 32 and a light chain having an amino acid sequence of SEQ ID NO: 34; and

[0347] (b) a second binding domain that specifically binds to TIGIT, the second binding domain comprising a heavy chain having an amino acid sequence of SEQ ID NO:42 and a light chain having an amino acid sequence of SEQ ID NO:44.

[0348] In another preferred embodiment, the bispecific binding protein comprises:

[0349] a) a first binding domain that specifically binds to PD-1, wherein the first binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises a CDRH1 having an amino acid sequence of SEQ ID NO: 52, a light chain variable domain having an amino acid sequence of SEQ ID NO:

[0350] The light chain variable domain comprises a CDRL1 having an amino acid sequence of SEQ ID NO:49, a CDRL2 having an amino acid sequence of SEQ ID NO:50, and a CDRL3 having an amino acid sequence of SEQ ID NO:51; and

[0351] b) a second binding domain that specifically binds to CTLA-4, wherein the second binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprising CDRH1 having an amino acid sequence of SEQ ID NO:58, CDRH2 having an amino acid sequence of SEQ ID NO:59, and CDRH3 having an amino acid sequence of SEQ ID NO:60, and the light chain variable domain comprising CDRL1 having an amino acid sequence of SEQ ID NO:55, CDRL2 having an amino acid sequence of SEQ ID NO:56

[0352] and CDRL3 having the amino acid sequence of SEQ ID NO:57.

[0353] More preferably, the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence of SEQ ID NO: 46 and a light chain having an amino acid sequence of SEQ ID NO: 45. In some aspects, 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 aspects, 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.

[0354] 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 aspects, 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 aspects, 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.

[0355] In a particularly preferred embodiment, the bispecific binding protein is an antibody comprising:

[0356] (a) a first binding domain that specifically binds to PD-1, the first binding domain comprising a heavy chain having an amino acid sequence of SEQ ID NO: 46 and a light chain having an amino acid sequence of SEQ ID NO: 45; and

[0357] (b) a second binding domain that specifically binds to CTLA-4, the second binding domain comprising 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.

[0358] In another particularly preferred embodiment, the bispecific binding protein is MEDI5752. As used herein, the term "MEDI5752" or "volrustomig" refers to an anti-PD-1 / CTLA-4 bispecific antibody comprising a light chain of SEQ ID NO: 45 and a heavy chain of SEQ ID NO: 46 (PD-1) and a light chain of SEQ ID NO: 47 and a heavy chain of SEQ ID NO: 48 (CTLA-4). MEDI5752 is disclosed in U.S. Pat. No. 10,457,732, which is incorporated herein by reference in its entirety.

[0359] In some aspects, the light chain constant region of the bispecific binding protein is a kappa chain. In some aspects, the light chain constant region of the bispecific binding protein is a lambda chain.

[0360] In some aspects, the bispecific binding protein comprises a variant Fc region comprising at least one substitution as numbered by the EU index as set forth in Kabat selected from the group consisting of 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, 24 1E, 241R, 243W, 243L, 243Y, 243R, 243Q, 244H, 245A, 247L, 247V, 247G, 250E, 250Q, 251F, 252L, 252Y, 254S, 254T, 255L, 256E, 256F, 256M, 257C, 257M, 25 7N, 262I, 262A, 262T, 262E, 263I, 263A, 263T, 263M, 264L, 264I, 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265A, 265G, 265N, 265Q, 265Y, 265F, 265V, 26 5I, 265L, 265H, 265T, 266I, 266A, 266T, 266M, 267Q, 267L, 268E, 269H, 269Y, 269F, 269R, 270E, 280A, 284M, 292P, 292L, 296E, 296Q, 296D, 296N, 296S, 29 6T, 296L, 296I, 296H, 296G, 297S, 297D, 297E, 298A, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 305I, 308F, 313F, 316D, 31 8A, 318S, 320A, 320S, 322A, 322S, 325Q, 325L, 3251, 325D, 325E, 325A, 325T, 325V, 325H, 326A, 326D, 326E, 326G, 326M, 326V, 327G, 327W, 327N, 327L, 32 8S, 328M, 328D, 328E, 328N, 328Q, 328F, 3281, 328V, 328T, 328H, 328A, 329F,329H, 329Q, 330K, 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.

[0361] In some aspects, the variant Fc region comprises one or more modifications at a position selected from 428 and 434 as numbered by the EU index as set forth in Kabat. In some aspects, the variant Fc region comprises one or more amino acid substitutions at a position selected from 428 and 434 as numbered by the EU index as set forth in Kabat. In some aspects, the variant Fc region comprises one or more amino acid substitutions selected from 428L, 428F, 434A, 434W, and 434Y.

[0362] Fc region engineering is widely used in the art to extend the half-life of therapeutic antibodies and prevent degradation in vivo. In some aspects, the Fc region of an IgG antibody or antigen binding fragment can be modified to increase the affinity of the IgG molecule for Fc receptor-neonate (FcRn), which mediates IgG catabolism and protects IgG molecules from degradation.

[0363] In some aspects, the antibody or antigen-binding fragment thereof comprises an Fc region that has been engineered to improve half-life.

[0364] In some aspects, the Fc region is non-glycosylated. In some aspects, the Fc region is deglycosylated. In some aspects, the Fc region has reduced fucosylation or is non-fucosylated.

[0365] In some aspects, the Fc variant antibody or binding fragment thereof has increased binding affinity for FcRn.

[0366] The triple mutation (TM) L234F / L235E / P331S in the heavy chain constant region (according to the EU numbering convention; Sazinsky et al., Proc Natl Acad Sci USA, 105:20167-20172 (2008)) can significantly reduce IgG effector function. In some aspects, the antibody or its antigen-binding fragment includes an Fc region with a triple mutation (TM) of L234F / L235E / P331S.

[0367] In some aspects, when used in vivo, Fc variant antibodies or their binding fragments have the complement dependent cytotoxicity (CDC) of reduction. In some aspects, compared to the antibody or its binding variants including wild-type Fc district, Fc variant antibodies or their binding fragments have the CDC of reduction. In some aspects, when used in vivo, Fc variant antibodies or their binding fragments do not trigger CDC. In some aspects, when used in vivo, Fc variant antibodies or their binding fragments cause the CDC of reduction. In some aspects, the Fc variant antibodies or their binding fragments with reduced CDC activity or without CDC activity include triple mutation (L234F / L235E / P331S) in variant Fc district.

[0368] In some aspects, when used in vivo, Fc variant antibodies or their binding fragments have reduced antibody-dependent cellular toxicity (ADCC). In some aspects, compared to the antibody or its binding variants including wild-type Fc district, Fc variant antibodies or their binding fragments have reduced ADCC. In some aspects, when used in vivo, Fc variant antibodies or their binding fragments do not trigger ADCC. In some aspects, when used in vivo, Fc variant antibodies or their binding fragments cause reduced ADCC. In some aspects, there is reduced ADCC activity or Fc variant antibodies or their binding fragments without ADCC activity include triple mutations (L234F / L235E / P331S) in variant Fc district.

[0369] In some aspects, when administered to a subject, an antibody or its binding fragment with reduced CDC activity has reduced toxicity. In some aspects, when administered to a subject, an antibody or its binding fragment with reduced ADCC activity has reduced toxicity.

[0370] As used herein, the term "Fc domain" includes native Fc and Fc variants and sequences as defined above. Like 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.

[0371] As used herein, the term "native Fc" refers to a molecule including a sequence of a non-antigen binding fragment produced by digestion of an antibody or produced by other means, whether in monomeric or multimeric form, and may include a hinge region. The original immunoglobulin source of natural Fc is preferably human, and may be any of immunoglobulins. Natural Fc molecules are composed of monomeric polypeptides, which can be connected to form dimers or multimeric forms by covalent (i.e., disulfide bonds) and non-covalent associations. The number of intermolecular disulfide bonds between the monomer subunits of natural Fc molecules is in the range of 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 natural Fc is a disulfide bonded dimer produced by papain digestion of IgG. As used herein, the term "native Fc" is a generic name for monomeric, dimer and multimeric forms.

[0372] In some aspects, the Fc region is or includes a domain from one or more of the Fc regions of IgGl, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD. In some aspects, the antibody is an IgGl antibody.

[0373] As used herein, the term "Fc variant" refers to a molecule or sequence modified from a native Fc but still including a binding site for the salvage receptor FcRn (neonatal Fc receptor). Exemplary Fc variants, and their interactions with salvage receptors are known in the art. Thus, the term "Fc variant" may include molecules or sequences humanized from non-human native Fc. In addition, native Fc includes regions that can be removed or mutated to produce Fc variants to alter certain residues that provide structural features or biological activity that are not required by the binding proteins of the present disclosure. Thus, the term "Fc variant" includes molecules or sequences that lack one or more native Fc sites or residues, or in which one or more Fc sites or residues have been modified, which affects or involves: (1) disulfide bond formation, (2) incompatibility with a selected host cell, (3) N-terminal heterogeneity when expressed in a selected host cell, (4) glycosylation, (5) interaction with complement, (6) binding to Fc receptors other than salvage receptors, or (7) antibody-dependent cellular cytotoxicity (ADCC).

[0374] In order to improve the yield of binding proteins, the CH3 domain can be altered by the "knob-into-holes" technique, which is described in detail using multiple exemplary molecules in, for example, 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 surfaces of the two CH3 domains are altered to increase the heterodimerization of the two heavy chains containing the two CH3 domains. Each of the two CH3 domains (of the two heavy chains) can be a "knob" and the other a "hole". The introduction of disulfide bridges further stabilizes the heterodimers (Merchant et al., 1998; Atwell et al., 1997, J. Mol. Biol. 270: 26-35) and improves the yield.

[0375] In some aspects, the binding protein has a "DuetMab" format. DuetMab has the following basic structure: an Fc region with a modified heavy chain, wherein the CH1 region of the modified heavy chain has a substitution of a natural cysteine ​​to a non-cysteine ​​amino acid, and a substitution of a natural non-cysteine ​​amino acid to a cysteine ​​amino acid; a modified corresponding light chain, wherein the CL region of the modified light chain also has a substitution of a natural cysteine ​​to a non-cysteine ​​amino acid, and a substitution of a natural non-cysteine ​​amino acid to a cysteine ​​amino acid; a second Fc region with a second heavy chain; and a second corresponding modified light chain, wherein the modified heavy chain is directly connected to the corresponding modified light chain, and on a separate target binding arm, the second heavy chain is directly connected to the second corresponding light chain, and wherein the substituted cysteine ​​of the modified heavy chain (generated by the substitution of a natural non-cysteine ​​amino acid to a cysteine ​​amino acid) and the substituted cysteine ​​of the modified corresponding light chain (generated by the substitution of a natural non-cysteine ​​amino acid to a cysteine ​​amino acid) can form a disulfide bond. Disclosure regarding DuetMab can be found, for example, in U.S. Pat. No. 9,527,927, which is incorporated herein by reference in its entirety.

[0376] As used herein, the term "KD" refers to the dissociation constant (KD = [A] x [B] / [AB]) of the interaction between the binding protein of the present disclosure and the antigen target, and the unit is mol / liter. The binding protein of the present disclosure generally has a dissociation constant (KD) of 10-5 mol / liter to 10-12 mol / liter or less, or 10-7 mol / liter to 10-12 mol / liter or less, or 10-3 mol / liter to 10-12 mol / liter, and / or has 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 mol / liter is generally considered to indicate non-specific binding. Therefore, the lower the KD value, the greater the affinity. In some aspects, the binding proteins of the present disclosure will 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 generally associated with higher potency, but it is not always the case that greater affinity means greater potency.

[0377] The dissociation constant (KD) can be determined, for example, by surface plasmon resonance (SPR). Typically, surface plasmon resonance analysis uses, for example The system (Pharmacia Biosensor; Piscataway, NJ) measures the real-time binding interaction (both association rate and dissociation rate) between the ligand (target antigen on the biosensor matrix) and the analyte by surface plasmon resonance. 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 radioimmunoassays (RIA), enzyme-linked immunosorbent assays (ELISA), enzyme immunoassays (EIA), and sandwich competition assays.

[0378] In one aspect, the equilibrium dissociation constant (KD) of the interaction between the bispecific binding protein as described herein and human TIGIT is less than or equal to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein as described herein and human TIGIT is less than or equal to about 9 pM. In one aspect, the KD of the interaction between the bispecific binding protein as described herein and human TIGIT is less than or equal to about 15 pM, 14 pM, 13 pM, 12 pM, 11 pM, 10 pM, 9 pM or 8 pM.

[0379] In one aspect, the KD of the interaction between the bispecific binding protein as described herein and human TIGIT is about 9 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein as described herein and human TIGIT is about 10 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein as described herein and human TIGIT is about 11 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein as described herein and human TIGIT is about 12 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein as described herein and human TIGIT is about 13 pM to about 15 pM. In one aspect, the KD of the interaction between the bispecific binding protein as described herein and human TIGIT is about 14 pM to about 15 pM.

[0380] In one aspect, the KD of the interaction of the bispecific binding protein as described herein with human PD-L1 is less than or equal to about 0.4 nM. In one aspect, the KD of the interaction of the bispecific binding protein as described herein with human PD-L1 is about 0.2 nM to about 0.5 nM. In one aspect, the KD of the interaction of the bispecific binding protein as described herein with human PD-L1 is about 0.3 nM to about 0.5 nM. In one aspect, the KD of the interaction of the bispecific binding protein as described herein with human PD-L1 is about 0.4 nM to about 0.5 nM.

[0381] 5. Combinations of Antibody-Drug Conjugates and Bispecific Checkpoint Inhibitors

[0382] In a first combination embodiment of the present disclosure, the antibody-drug conjugate combined with a bispecific checkpoint inhibitor is an antibody-drug conjugate in which the antibody is an anti-TROP2 antibody.

[0383] In one embodiment of the first combination embodiment described above, the anti-TROP2 antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises CDRH1 consisting of an amino acid sequence represented by SEQ ID NO:3 [= amino acid residues 50 to 54 of SEQ ID NO:1], CDRH2 consisting of an amino acid sequence represented by SEQ ID NO:4 [= amino acid residues 69 to 85 of SEQ ID NO:1], and CDRH3 consisting of an amino acid sequence represented by SEQ ID NO:5 [= amino acid residues 118 to 129 of SEQ ID NO:1], and the light chain comprises CDRL1 consisting of an amino acid sequence represented by SEQ ID NO:6 [= amino acid residues 44 to 54 of SEQ ID NO:2], CDRL2 consisting of an amino acid sequence represented by SEQ ID NO:7 [= amino acid residues 70 to 76 of SEQ ID NO:2], and CDRL3 consisting of an amino acid sequence represented by SEQ ID NO:8 [= amino acid residues 109 to 117 of SEQ ID NO:2]. In another embodiment of the first combination embodiment described above, the anti-TROP2 antibody comprises a heavy chain comprising a heavy chain variable region consisting of an amino acid sequence represented by SEQ ID NO:9 [=amino acid residues 20 to 140 of SEQ ID NO:1], and a light chain comprising a light chain variable region consisting of an amino acid sequence represented by SEQ ID NO:10 [=amino acid residues 21 to 129 of SEQ ID NO:2]. In another embodiment of the first combination embodiment described above, the anti-TROP2 antibody comprises a heavy chain consisting of an 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 an amino acid sequence represented by SEQ ID NO:13 [=amino acid residues 21 to 234 of SEQ ID NO:2]. In another embodiment of the first combination embodiment described above, the anti-TROP2 antibody comprises a heavy chain consisting of an amino acid sequence represented by SEQ ID NO: 11 [= amino acid residues 20 to 469 of SEQ ID NO: 1] and a light chain consisting of an amino acid sequence represented by SEQ ID NO: 13 [= amino acid residues 21 to 234 of SEQ ID NO: 2]. In another embodiment of the first combination embodiment described above, the anti-TROP2 is deludecanoic acid (DS-1062).

[0384] In a second combination embodiment of the present disclosure, the antibody-drug conjugate combined with the bispecific check point inhibitor is an antibody-drug conjugate wherein the antibody is an anti-HER2 antibody.

[0385] In one embodiment of the second combination embodiment described above, the anti-HER2 antibody comprises a heavy chain and a light chain, the 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 the 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 and a light chain, the heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22, and the 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 above second combination embodiment, 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 above second combination embodiment, the anti-HER2 antibody is trastuzumab (DS-8201).

[0386] In a particularly preferred embodiment of the above-mentioned first combination embodiment, the anti-TROP2 antibody-drug conjugate is derutec (DS-1062), and the bispecific checkpoint inhibitor is an anti-PD-1 / TIGIT bispecific antibody, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence of SEQ ID NO:32 and a light chain having an amino acid sequence of SEQ ID NO:34, and wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence of SEQ ID NO:41 and a light chain variable domain having an amino acid sequence of SEQ ID NO:43.

[0387] In another particularly preferred embodiment of the above-mentioned first combination embodiment, the anti-TROP2 antibody-drug conjugate is derutec (DS-1062), and the bispecific checkpoint inhibitor is an anti-PD-1 / CTLA-4 bispecific antibody, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence of SEQ ID NO:46 and a light chain having an amino acid sequence of SEQ ID NO:45, and wherein the second binding domain that specifically binds to CTLA-4 comprises a heavy chain variable domain having an amino acid sequence of SEQ ID NO:48 and a light chain variable domain having an amino acid sequence of SEQ ID NO:47.

[0388] In a particularly preferred embodiment of the above-mentioned second combination embodiment, the anti-HER2 antibody-drug conjugate is trastuzumab (DS-8201), and the bispecific checkpoint inhibitor is an anti-PD-1 / TIGIT bispecific antibody, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence of SEQ ID NO: 32 and a light chain having an amino acid sequence of SEQ ID NO: 34, and wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence of SEQ ID NO: 41 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 43.

[0389] In another particularly preferred embodiment of the above-mentioned second combination embodiment, the anti-HER2 antibody-drug conjugate is trastuzumab (DS-8201), and the bispecific check point inhibitor is an anti-PD-1 / CTLA-4 bispecific antibody, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence of SEQ ID NO:46 and a light chain having an amino acid sequence of SEQ ID NO:45, and wherein the second binding domain that specifically binds to CTLA-4 comprises a heavy chain variable domain having an amino acid sequence of SEQ ID NO:48 and a light chain variable domain having an amino acid sequence of SEQ ID NO:47.

[0390] In some aspects of the above-mentioned combination embodiments, the antibody-drug conjugate and the bispecific checkpoint inhibitor are further co-administered 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).

[0391] 6. Therapeutic Combination Uses and Methods

[0392] Described below is a pharmaceutical product and therapeutic uses and methods in which an antibody-drug conjugate according to the present disclosure (preferably an anti-TROP2 or anti-HER2 antibody-drug conjugate) and a bispecific checkpoint inhibitor are administered in combination.

[0393] The pharmaceutical products and therapeutic uses and methods of the present disclosure may be characterized in that the antibody-drug conjugate and the bispecific check point inhibitor are contained as active ingredients in separate formulations and administered simultaneously or at different times, or may be characterized in that the antibody-drug conjugate and the bispecific check point inhibitor are contained as active ingredients in a single formulation and administered.

[0394] In the pharmaceutical products and treatment methods of the present disclosure, a single bispecific check point inhibitor used in the present disclosure can be administered in combination with an antibody-drug conjugate, or two or more different bispecific check point inhibitors can be administered in combination with an antibody-drug conjugate.

[0395] The pharmaceutical products and treatment methods of the present disclosure can be used to treat cancer, and can preferably be used to treat at least one cancer 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 carcinoma, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumors, digestive tract stromal tumors, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, uterine corpus cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasma cell tumor, myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer and renal cell carcinoma.

[0396] The presence or absence of tumor markers such as HER2 or TROP2 can be determined, for example, by collecting tumor tissue from a cancer patient to prepare a formalin-fixed paraffin-embedded (FFPE) specimen and testing the specimen for gene products (proteins) using, for example, immunohistochemistry (IHC), flow cytometry or western blotting, or testing the specimen for gene transcription using in situ hybridization (ISH), quantitative PCR (q-PCR) or microarray analysis, or by collecting cell-free circulating tumor DNA (ctDNA) from a cancer patient and testing the ctDNA by a method such as next-generation sequencing (NGS).

[0397] The drug products and treatment methods of the present disclosure, when comprising an anti-HER2 antibody-drug conjugate, can be used for cancers that express HER2, which can be cancers that overexpress HER2 (high or intermediate) or can be cancers that underexpress HER2.

[0398] In the present disclosure, the term "cancer overexpressing HER2" is not particularly limited as long as it is identified as a cancer overexpressing HER2 by a person skilled in the art. Preferred examples of cancer overexpressing HER2 may include cancers with a score of 3+ for HER2 expression in the IHC method, and cancers with a score of 2+ for HER2 expression in the IHC method and determined to be HER2 expression-positive in an in situ hybridization method (ISH). The in situ hybridization method disclosed herein includes a fluorescent in situ hybridization method (FISH) and a dual-color in situ hybridization method (DISH).

[0399] In the present disclosure, the term "cancer that lowly expresses HER2" is not particularly limited as long as it is recognized as a cancer that lowly expresses HER2 by a person skilled in the art. Preferred examples of cancer that lowly expresses HER2 may include cancers that score 2+ for HER2 expression in the IHC method and are determined to be negative for HER2 expression in the in situ hybridization method, and cancers that score 1+ for HER2 expression in the IHC method.

[0400] The method for scoring the degree of HER2 expression by IHC method, or determining the positive or negative method of HER2 expression by in situ hybridization method is not particularly limited, as long as it is recognized by those skilled in the art. The example of the method may include the method described in the 4th edition of the breast cancer HER2 test guide (developed by the Japanese Pathology Board for Optimal Use of HER2 for Breast Cancer).

[0401] The cancer, particularly with respect to the treatment of breast cancer, can be breast cancer that overexpresses HER2 (high or moderate) or underexpresses HER2, or triple-negative breast cancer, and / or can have a HER2 status score of IHC 3+, IHC 2+, IHC 1+, or IHC>0 and <1+.

[0402] In some aspects, the methods of the present disclosure include identifying that the patient has a PD-L1 positive tumor prior to administering the bispecific checkpoint inhibitor.

[0403] In some aspects, a PD-L1 positive tumor comprises 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% of cells expressing PD-L1.

[0404] In some aspects, PD-L1 expression is determined by receiving the results of an assay capable of determining PD-L1 expression. In some aspects, the assay capable of determining PD-L1 expression is the Ventana PD-L1 (SP263) IHC assay, the 22C3 PharmDx assay, or the 28-8PharmDx assay.

[0405] In some aspects, the methods of the present disclosure comprise administering an antibody-drug conjugate and an anti-PD-1 / CTLA-4 bispecific antibody to a patient whose tumor contains less than 50% of cells expressing PD-L1. In some aspects, the methods of the present disclosure comprise administering an antibody-drug conjugate, an anti-PD-1 / CTLA-4 bispecific antibody, and carboplatin to a patient whose tumor contains less than 50% of cells expressing PD-L1.

[0406] In some aspects, the methods of the present disclosure comprise administering an antibody-drug conjugate and an anti-PD-1 / TIGIT bispecific antibody to a patient whose tumor contains at least 50% cells expressing PD-L1. In some aspects, the methods of the present disclosure comprise administering an antibody-drug conjugate, an anti-PD-1 / TIGIT bispecific antibody, and carboplatin to a patient whose tumor contains less than 50% cells expressing PD-L1.

[0407] The pharmaceutical products and methods of treatment of the present disclosure may preferably be used in mammals, but more preferably in humans.

[0408] The anti-tumor effect of the drug product and treatment method of the present disclosure can be confirmed by transplanting cancer cells into a test subject animal to prepare a model and measuring the tumor volume reduction or life extension effect by administering the drug product and treatment method of the present disclosure. Then, the effect of the combined use of the antibody-drug conjugate used in the present disclosure and the bispecific checkpoint inhibitor can be confirmed by comparing the anti-tumor effect of administering the antibody-drug conjugate used in the present disclosure alone with the anti-tumor effect of administering the bispecific checkpoint inhibitor alone.

[0409] The anti-tumor effects of the pharmaceutical products and treatment methods disclosed herein can be confirmed in clinical trials using any of the evaluation methods with solid tumor response evaluation criteria (RECIST), WHO evaluation methods, Macdonald evaluation methods, body weight measurement, and other methods, and can be determined based on indicators such as complete remission (CR), partial remission (PR); disease progression (PD), objective response rate (ORR), duration of remission (DoR), progression-free survival (PFS), overall survival (OS), etc.

[0410] By using the above method, it can be confirmed that the anti-tumor effect of the pharmaceutical product and treatment method disclosed in the present invention is superior to the existing pharmaceutical products and treatment methods for cancer treatment.

[0411] The pharmaceutical product and the method of treatment disclosed herein can delay the development of cancer cells, inhibit their growth, and further kill cancer cells. These effects can make cancer patients get rid of the symptoms caused by cancer or realize the improvement of the quality of life (QOL) of cancer patients, and realize the therapeutic effect by maintaining the life of cancer patients. Even if the pharmaceutical product and the method of treatment disclosed herein cannot realize killing cancer cells, they can also realize the higher QOL of cancer patients by inhibiting or controlling the growth of cancer cells, and realize longer-term survival.

[0412] The pharmaceutical products of the present disclosure can be expected to exert therapeutic effects by administration to the patient as systemic therapy, and additionally by local administration to cancer tissue.

[0413] In another aspect, the pharmaceutical products and treatment methods disclosed herein provide for use as adjuvants in cancer therapy using ionizing radiation or other chemotherapeutic agents. For example, in the treatment of cancer, treatment may include administering a therapeutically effective amount of the pharmaceutical product to a subject in need of treatment, while administering ionizing radiation or other chemotherapeutic agents, or sequentially.

[0414] The pharmaceutical products and treatment methods of the present disclosure can be used as adjuvant chemotherapy in combination with surgery. The pharmaceutical products of the present disclosure can be administered before surgery for the purpose of reducing tumor size (known as preoperative adjuvant chemotherapy or neoadjuvant therapy), or can be administered after surgery for the purpose of preventing tumor recurrence (known as postoperative adjuvant chemotherapy or adjuvant therapy).

[0415] In some embodiments, the cancer cell may have a BRCA1 and / or BRCA2 defective phenotype, i.e., BRCA1 and / or BRCA2 activity is reduced or eliminated in the cancer cell. Cancer cells with this phenotype may lack BRCA1 and / or BRCA2, i.e., the expression and / or activity of BRCA1 and / or BRCA2 may be reduced or eliminated in the cancer cell, for example, by mutation or polymorphism in the encoding nucleic acid, or by amplification, mutation or polymorphism in a gene encoding a regulatory factor (e.g., 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 often lost in tumors of heterozygous carriers (Jasin M., Oncogene, 21 (58), 8981-93 (2002); Tutt et al., Trends Mol Med., 8 (12), 571-6, (2002)). The association of BRCA1 and / or BRCA2 mutations with breast cancer is well characterized in the art (Radice, PJ, Exp Clin Cancer Res., 21(3 Suppl), 9-12 (2002)). Amplification of the EMSY gene, which encodes a BRCA2 binding factor, is also known to be associated with breast and ovarian cancer. Carriers of mutations in BRCA1 and / or BRCA2 are also at high risk for certain cancers, including breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, blood cancer, gastrointestinal cancer, and lung cancer. In some embodiments, the individual is heterozygous for one or more variations (such as mutations and polymorphisms) in BRCA1 and / or BRCA2 or its regulatory factors. Detection of mutations in BRCA1 and BRCA2 is well known in the art and is described, for example, in EP 699754, EP 705903, Neuhausen, SL and Ostrander, EA, Genet. Test, 1, 75-83 (1992); Chappnis, PO and Foulkes, WO, 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 amplification of the BRCA2 binding factor EMSY is described in Hughes-Davies et al., Cell, 115, 523-535.

[0416] Cancer-associated mutations and polymorphisms can be detected at the nucleic acid level by detecting the presence of variant nucleic acid sequences, or at the protein level by detecting the presence of variant (ie, mutant or allelic variant) polypeptides.

[0417] The drug product of the present disclosure containing at least one pharmaceutically suitable ingredient can be administered. According to the dosage, administration concentration, etc. of the antibody-drug conjugate and bispecific checkpoint inhibitor used in the present disclosure, the pharmaceutically suitable ingredient can be appropriately selected and applied from the formulation additives commonly used in the art, etc. The antibody-drug conjugate used in the present disclosure can be administered, for example, as a drug product containing 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 drug product of the present disclosure can be preferably used as an injection, can be more preferably used as an aqueous injection or a lyophilized injection, and can be even more preferably used as a lyophilized injection. In the case where the drug product containing the antibody-drug conjugate used in the present disclosure is an aqueous injection, the aqueous injection can preferably be diluted with a suitable diluent and then administered in the form of intravenous infusion. Examples of diluents can include glucose solutions and saline solutions, and glucose solutions can be preferably exemplified, and 5% glucose solutions can be more preferably exemplified. In the case where the drug product of the present disclosure is a lyophilized injection, the lyophilized injection of the required amount pre-dissolved in water for injection can be preferably diluted with a suitable diluent and then administered in the form of intravenous infusion. Examples of diluents can include glucose solution and physiological saline, glucose solution can be preferably exemplified, and 5% glucose solution can be more preferably exemplified.

[0418] Examples of administration routes suitable for administering the drug product of the present disclosure may include intravenous, intradermal, subcutaneous, intramuscular, and intraperitoneal routes, with the intravenous route being preferred.

[0419] The size of the dose required for treating a particular disease state will necessarily vary depending on the subject being treated, the route of administration, and the severity of the disease being treated. For further information on routes of administration and dosage regimens, reference can be made to Volume 5, Chapter 25.3 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.

[0420] The anti-TROP2 antibody-drug conjugate used in the present disclosure can be administered to humans once at intervals of 1 day to 180 days, and can preferably be administered once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks, and can even more preferably be administered once every 3 weeks. In addition, the antibody-drug conjugate used in the present disclosure can be administered at a dose of about 0.001 mg / kg to 100 mg / kg, and can preferably be administered at a dose of 0.8 mg / kg to 12.4 mg / kg. For example, the anti-TROP2 antibody-drug conjugate can be administered once every 3 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, and can preferably be administered once every 3 weeks at a dose of 4.0 mg / kg or 6.0 mg / kg.

[0421] The anti-HER2 antibody-drug conjugate used in the present disclosure can be applied to humans at intervals of 1 day to 180 days, preferably at intervals of one week, two weeks, three weeks or four weeks, and more preferably at intervals of three weeks. The anti-HER2 antibody-drug conjugate used in the present disclosure can be applied at a dosage of about 0.001 mg / kg to 100 mg / kg per application, and can preferably be applied at a dosage of 0.8 mg / kg to 12.4 mg / kg per application. For example, the anti-HER2 antibody-drug conjugate can be applied once every three weeks at a dosage 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, and can preferably be applied once every three weeks at a dosage of 5.4 mg / kg or 6.4 mg / kg.

[0422] The bispecific checkpoint inhibitors can be administered by any suitable route of administration at a suitable dose.

[0423] In some aspects, 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 aspects, the administered dose 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 1110 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 1210 mg, about 1220 mg, about 1230 mg, about 1240 mg, about 1250 mg, about 1260 mg, about 1270 mg, about 1280 mg, about 1290 mg, about 1300 mg, about 1310 mg, about 1320 mg, about 1330 mg, about 1340 mg, about 1350 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.

[0424] In some aspects, a dose of MEDI5752 or an antigen binding fragment thereof is administered to the subject once per treatment cycle. In some aspects, the treatment cycle is three weeks. In some aspects, a 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.

[0425] In some aspects, the bispecific antibody or its antigen-binding fragment is co-administered 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). In some aspects, the chemotherapeutic agent is pemetrexed. In some aspects, the chemotherapeutic agent is axitinib.

[0426] In some aspects, the bispecific checkpoint inhibitors disclosed herein (such as binding proteins) 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 of the routes of administration using methods known in the art. The term "pharmaceutically acceptable carrier" means one or more non-toxic materials that do not interfere with the effectiveness of the biological activity of the active ingredient. Such preparations may conventionally include salts, buffers, preservatives, compatible carriers and optional other therapeutic agents. Such pharmaceutically acceptable preparations may also include compatible solid or liquid fillers, diluents or encapsulating materials suitable for administration to humans. Other expected carriers, excipients and / or additives that may be utilized in the preparations described herein include, for example, flavorings, antimicrobial agents, sweeteners, antioxidants, antistatic agents, lipids, protein excipients such as serum albumin, gelatin, casein, salt-forming counterions such as sodium, etc. These and additional known pharmaceutical carriers, excipients and / or additives suitable for use in the formulations described herein are known in the art, for example, as listed in "Remington: The Science & Practice of Pharmacy", 21st edition, Lippincott Williams & Wilkins, (2005) and "Physician's Desk Reference", 60th edition, Medical Economics, Montvale, NJ (2005). A pharmaceutically acceptable carrier may be selected to be suitable for the desired or required mode of administration, solubility and / or stability.

[0427] In some respects, therapeutic compositions can be formulated for specific routes of administration, such as oral, nasal, pulmonary, local (including oral and sublingual), rectal, vaginal and / or parenteral administration. As used herein, the terms "parenteral administration" and "parenteral administration" refer to modes of administration except 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, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, supradalateric and intrasternal injection and infusion. Preparations of the present disclosure suitable for topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The antibodies and other active substances may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required (see, e.g., U.S. Patents 7,378,110; 7,258,873; and 7,135,180; U.S. Patent Application Publications 2004 / 0042972 and 2004 / 0042971).

[0428] Preparations can exist in unit dosage form and can be prepared by any method known in the pharmaceutical field. The actual dosage level of the active ingredient in the pharmaceutical composition of the present disclosure can be varied, so as to obtain the amount of active ingredient (e.g., "therapeutically effective amount") that effectively achieves the desired therapeutic response for a specific patient, composition, and mode of administration without toxicity to the patient. The selected dosage level will depend on a variety of pharmacokinetic factors, including the activity of the specific composition used, the route of administration, the time of administration, the excretion rate of the specific compound used, the duration of treatment, other drugs, compounds and / or materials used in combination with the specific composition used, the age, sex, body weight, condition, general health and previous medical history of the patient treated, and similar factors well known in the medical field. These dosages can be applied daily, weekly, biweekly, monthly or at a lower frequency (e.g., every six months), depending on dosage, method of administration, illness or symptom to be treated, and individual subject characteristics. Dosage can also be applied via continuous infusion (such as by pump). The dosage applied can also depend on the route of administration. For example, subcutaneous administration may require a higher dosage than intravenous administration. As described above, any commonly used dosing regimen (eg, 1 mg / kg-10 mg / kg, administered daily or twice weekly by injection or infusion) can be adapted and used in connection with the methods of treating human cancer patients.

[0429] [Example]

[0430] The present disclosure is specifically described in view of the embodiments shown below. However, the present disclosure is not limited to these. In addition, it is by no means interpreted in a limiting manner.

[0431] Example 1A: Preparation of anti-TROP2 antibody-drug conjugates

[0432] According to the preparation methods described in WO 2015 / 098099 and WO 2017 / 002776 and using an anti-TROP2 antibody (an antibody comprising a heavy chain consisting of an 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 an amino acid sequence represented by SEQ ID NO: 13 [= amino acid residues 21 to 234 of SEQ ID NO: 2]), an anti-TROP2 antibody-drug conjugate in which a drug-linker represented by the following formula is conjugated to the anti-TROP2 antibody via a thioether bond was prepared:

[0433]

[0434] Where A represents the attachment position to the antibody (DS-1062: derutecan). The DAR of the antibody-drug conjugate is about 4.

[0435] Example 1B: Preparation of anti-HER2 antibody-drug conjugates

[0436] According to the preparation method described in WO2015 / 115091 and using an anti-HER2 antibody (an antibody comprising a heavy chain consisting of an 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 an amino acid sequence consisting of all amino acid residues 1 to 214 of SEQ ID NO: 15), an anti-HER2 antibody-drug conjugate in which a drug-linker represented by the following formula is conjugated to the anti-HER2 antibody via a thioether bond was prepared:

[0437]

[0438] Where A represents the attachment position to the antibody (DS-8201: trastuzumab). The DAR of the antibody-drug conjugate is 7.7 or 7.8.

[0439] Example 2A: Anti-PD-1 / TIGIT Bispecific Antibodies (Bispecific Checkpoint Inhibitors)

[0440] An anti-PD-1 / TIGIT bispecific antibody (monovalent, humanized, IgG1 monoclonal) was produced having a first binding domain that specifically binds to PD-1, the first binding domain comprising a heavy chain having an amino acid sequence of SEQ ID NO: 32 and a light chain having an amino acid sequence of SEQ ID NO: 34, and a second binding domain that specifically binds to TIGIT, the second binding domain comprising a heavy chain variable domain having an amino acid sequence of SEQ ID NO: 42 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 44. The antibody was engineered to have a triple mutation L234F / L235E / P331S within its Fc domain to reduce Fc-mediated effector function.

[0441] Example 2B: Anti-PD-1 / CTLA-4 Bispecific Antibody (Bispecific Checkpoint Inhibitor)

[0442] The anti-PD-1 / CTLA-4 bispecific antibody MEDI5752 as disclosed in U.S. Pat. No. 10,457,732 was produced. MEDI5752 has a first binding domain that specifically binds to PD-1 and a second binding domain that specifically binds to CTLA-4, the first binding domain comprising a heavy chain having an amino acid sequence of SEQ ID NO: 46 and a light chain having an amino acid sequence of SEQ ID NO: 45, and the second binding domain comprising a heavy chain variable domain having an amino acid sequence of SEQ ID NO: 48 and a light chain variable domain having an amino acid sequence of SEQ ID NO: 47.

[0443] Example 3: Anti-tumor test

[0444] Combination of antibody-drug conjugate DS-8201 with anti-PD-L1 + anti-CTLA-4 or anti-PD-1 / TIGIT DuetMab

[0445] (i) Methods :

[0446] Tumor Models

[0447] Female BALB / c mice aged 7-9 weeks were purchased from Envigo and acclimated for at least 7 days before entering the study. For tumor implantation, the right flank of the mice was shaved and injected subcutaneously with 100 μL of 5 × 10 6 EMT6 human HER2 (hHER2) cells. Tumor volume was measured three times per week using an electronic caliper and the formula (width 2 × length) / 2.

[0448] Seven days after cell implantation, when the tumor reached approximately 170 mm 3Tumors of similar size were randomly assigned to treatment groups as shown in Table 1. Mice were euthanized when they reached humane welfare limits related to tumor volume (mean diameter 15 mm) or tumor condition (skin ulceration overlying the tumor).

[0449] CD34+ umbilical cord blood stem cells were transplanted into humanized female NSG mice using 3 different donors at Jackson Laboratory. Approximately 15 weeks after implantation, 1e6 Caki-1 cells were injected subcutaneously into the right flank. Tumor volume was measured twice weekly using a caliper and calculated using the formula (width 2 × length) / 2. When the tumor reaches 115 mm 3 When the mean volume of the tumor was determined, the mice were randomized into treatment groups as shown in Table 2. Mice were randomized by tumor volume and cord blood donor to ensure 3 donors per group.

[0450] Table 1

[0451]

[0452]

[0453] Table 2

[0454]

[0455] Preparation

[0456] Preparation of 10mg / kg DS-8201

[0457] The dosing solution of DS-8201 (HA306) was prepared by diluting the DS-8201 stock solution (20.1 mg / mL) to 2.5 mg / mL in ABS buffer (10 mM sodium acetate, 10 mM acetic acid, 5% sorbitol) and then administered via IV injection at a dosing volume of 4 mL / kg.

[0458] 10mg / kg Anti-PD-L1 Preparation

[0459] The dosing solution of anti-PD-L1 (clone 80, SP21-095, IgG1, D265A) was prepared by diluting the stock solution (11.2 mg / mL) to 1 mg / mL in PBS and then administered via IP injection at a dosing volume of 10 mL / kg.

[0460] 10mg / kg co-formulation of anti-PD-L1 and anti-CTLA-4

[0461] Dosing solutions of anti-PD-L1 (clone 80, SP21-095, IgG1, D265A) and anti-CTLA-4 (SP20-103, murine IgG1) were prepared by diluting stock solutions (anti-PD-L1–11.2 mg / mL; anti-CTLA-4–10 mg / mL) and co-formulating at 1 mg / mL of each agent in PBS and then administered via IP injection at a dose volume of 10 mL / kg.

[0462] Formulation of Anti-PD-1 / TIGIT DuetMab

[0463] The dosing solution of anti-PD-1 / TIGIT DuetMab (mIgG2a LALA-PG) was prepared by diluting the stock solution (C428223DEC21EO, 2.7 mg / mL) to 1 mg / mL in PBS and then administered via IP injection at a dosing volume of 10 mL / kg.

[0464] Formulation of MEDI5752

[0465] The dosing solution of MEDI5752 was prepared by diluting the stock solution (lot ML00669-50, 63.41 mg / ml) in PBS to 2 mg / ml and then administered via IP injection at a dosing volume of 5 mL / kg.

[0466] Fine Needle Aspiration (FNA) Sampling

[0467] The mouse was anesthetized using 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 the tissue. Negative pressure was generated by pulling the syringe plunger back about 100 μL to draw the cells into the medium. About 200 μL of medium containing cells was then flushed from the syringe into a 1.5 mL Eppendorf tube. This step was repeated four more times at different locations around the entire tumor to maximize the representativeness of the sampling, and the tube was immediately placed on ice.

[0468] Flow cytometry staining

[0469] Cells were stained with a fixable viability dye (Thermo Fisher) and blocked with antibodies to CD16 / CD32 (eBioscience) and then stained with fluorescently conjugated antibodies in flow cytometry staining buffer (2% bovine serum albumin, 0.1% sodium azide, 2mM EDTA) with 50% Brilliant staining buffer (BD Biosciences). Intracellular staining was performed using the FoxP3 / transcription factor staining buffer set (eBioscience) and 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). Cells containing less than 1000 CD45 + Samples of cells were excluded from analysis due to low event counts.

[0470] statistics

[0471] The tumor growth rate for each animal was calculated based on fitting the growth curve of each tumor (days 0-15 after the start of treatment) to the exponential model log10(tumor volume) = a + b·time + error, where a and b are parameters corresponding to the logarithmic initial volume and growth rate, respectively. The MannWhitney U test was used to evaluate the statistical significance of differences in tumor growth rate between groups. The box plots show the median and the 25th–75th quartiles, and the whiskers show the minimum and maximum values.

[0472] For flow cytometry data of cell frequency, data were processed using beta regression as fixed effect. p-values ​​are reported as follows; p ≥ 0.05 (ns, not significant); *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

[0473] (ii) Testing :

[0474] Mice bearing EMT6 hHER2 tumors were treated with a combination of DS-8201 and immuno-oncology (IO) agents, i.e., 10 mg / kg DS-8201 alone or in combination with 10 mg / kg anti-PD-L1, 10 mg / kg anti-PD-L1 + 10 mg / kg anti-CTLA-4, and 10 mg / kg anti-PD-1 / TIGIT DuetMab, according to the dosing schedule outlined in Table 1. Fig.59 and Fig.60 and in Tables 3 and 4.

[0475] According to the dosing regimen outlined in Table 1, FNA tumor samples were collected from mice bearing EMT6hHER2 tumors treated with DS-8201 alone or in combination with 10 mg / kg anti-PD-1 / TIGITDuetMab 8 days after the start of treatment, 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. Results are shown in Fig.61 In a similar study, tumor samples were collected from mice bearing EMT6hHER2 tumors treated with vehicle alone or DS-8201 10 days after the start of treatment, 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 middle.

[0476] 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. Results are shown in Fig.63 middle.

[0477] Peripheral blood was sampled on day 22 after treatment initiation in the Caki-1 study and analyzed by flow cytometry for human-specific T cell activation markers (n=5 samples / group). Fig.64 middle.

[0478] (iii) Results :

[0479] Fig.59 and Fig.60 The antitumor efficacy and tumor growth rate of the combination of DS-8201 and IO agents in mice bearing EMT6hHER2 tumors are shown, respectively. Fig.59 Figure 3 shows the mean changes in tumor volume over time. Data shown are mean ± SEM (n = 10 mice / treatment group). Fig.59 In Figure 2, panel B shows the changes in tumor volume over time in individual mice (CR = complete remission). Fig.60 In the Figures, data shown are tumor growth rates (n=10 mice / treatment group).

[0480] Single-agent DS-8201 treatment significantly delayed tumor growth (53.2% growth rate inhibition, p=0.0002), as did 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). Fig.59 Figure A, Fig.60 , Table 3).

[0481] Table 3

[0482]

[0483] 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), tumor growth inhibition was further enhanced compared with DS-8201 monotherapy ( Fig.59 The combination of DS-8201 with anti-PD-L1+anti-CTLA-4 and anti-PD-1 / TIGIT DuetMab resulted in complete remission in 80% and 60% of animals, respectively, whereas the combination with anti-PD-L1 resulted in complete remission in only 10% of animals ( Fig.59 Figure B, Table 4).

[0484] Table 4

[0485]

[0486] Regarding pharmacodynamic changes, FNA tumor samples collected 8 days after the start of DS-8201 treatment showed an increase in the tumor proportion of total immune cells (CD45+), Treg and NK cells, and an increase in the proportion of T cells expressing PD-1, Ki67 and TIGIT. Compared with monotherapy, the combination of DS-8201 and anti-PD-1 / TIGIT DuetMab treatment further increased the tumor T cell (survival %) and NK cell (CD45+%) content, and also increased the proportion of CD8+ T cells (CD3+%) ( Fig.61 In similar studies, DS-8201 monotherapy treatment significantly increased CTLA-4 expression on tumor CD8+ T cells ( Fig.62 ).

[0487] In a humanized mouse model bearing renal Caki-1 tumors transplanted with CD34+ hematopoietic stem cells, the combination of DS-8201 and MEDI5752 resulted in greater tumor growth inhibition than either monotherapy alone and was significantly superior to DS-8201 ( Fig.63 ). Mice receiving MEDI5752 monotherapy exhibited significant weight loss relative to the DS-8201 treatment group, which led to early termination of this treatment group at day 50 after tumor implantation. Interestingly, these effects were not observed in the group receiving MEDI5752 in combination with DS-8201.

[0488] In the Caki-1 study, peripheral blood flow analysis was performed on day 22 after treatment and revealed an increase in proliferating CD4+ and CD8+ T cells as well as an increase in granzyme B+CD8 and ICOS+CD4 cells. These effects were observed with MEDI5752 monotherapy and were maintained in the combination treatment group ( Fig.64 ).

[0489] The above written description is considered sufficient to enable those skilled in the art to practice these embodiments. The above description and examples detail certain embodiments and describe the best mode contemplated by the inventors. However, it should be understood that no matter how detailed the above content may appear in text, the embodiments may be practiced in many ways, and the claims include any equivalents thereof.

[0490] Free text for sequence listings

[0491] SEQ ID NO: 1—Amino acid sequence of the heavy chain of an anti-TROP2 antibody

[0492] SEQ ID NO:2—Amino acid sequence of the light chain of an anti-TROP2 antibody

[0493] SEQ ID NO:3—Amino acid sequence of heavy chain CDRH1 [=amino acid residues 50 to 54 of SEQ ID NO:1]

[0494] SEQ ID NO:4—Amino acid sequence of heavy chain CDRH2 [=amino acid residues 69 to 85 of SEQ ID NO:1]

[0495] SEQ ID NO:5—Amino acid sequence of heavy chain CDRH3 [=amino acid residues 118 to 129 of SEQ ID NO:1]

[0496] SEQ ID NO:6—Amino acid sequence of light chain CDRL1 [=amino acid residues 44 to 54 of SEQ ID NO:2]

[0497] SEQ ID NO:7—Amino acid sequence of light chain CDRL2 [=amino acid residues 70 to 76 of SEQ ID NO:2]

[0498] SEQ ID NO:8—Amino acid sequence of light chain CDRL3 [=amino acid residues 109 to 117 of SEQ ID NO:2]

[0499] SEQ ID NO:9—Amino acid sequence of the heavy chain variable region [=amino acid residues 20 to 140 of SEQ ID NO:1]

[0500] SEQ ID NO: 10—Amino acid sequence of the light chain variable region [=amino acid residues 21 to 129 of SEQ ID NO: 2]

[0501] SEQ ID NO: 11 - amino acid sequence of the heavy chain [= amino acid residues 20 to 469 of SEQ ID NO: 1]

[0502] SEQ ID NO: 12—Amino acid sequence of the heavy chain [=amino acid residues 20 to 470 of SEQ ID NO: 1]

[0503] SEQ ID NO: 13—Amino acid sequence of light chain [=amino acid residues 21 to 234 of SEQ ID NO: 2]

[0504] SEQ ID NO: 14 - Amino acid sequence of the heavy chain of an anti-HER2 antibody

[0505] SEQ ID NO: 15 - Amino acid sequence of the light chain of an anti-HER2 antibody

[0506] SEQ ID NO: 16—Amino acid sequence of heavy chain CDRH1 [=amino acid residues 26 to 33 of SEQ ID NO: 14]

[0507] SEQ ID NO: 17—Amino acid sequence of heavy chain CDRH2 [=amino acid residues 51 to 58 of SEQ ID NO: 14]

[0508] SEQ ID NO: 18—Amino acid sequence of heavy chain CDRH3 [=amino acid residues 97 to 109 of SEQ ID NO: 14]

[0509] SEQ ID NO: 19—Amino acid sequence of light chain CDRL1 [=amino acid residues 27 to 32 of SEQ ID NO: 15]

[0510] SEQ ID NO:20—Amino acid sequence comprising the amino acid sequence of light chain CDRL2 (SAS) [=amino acid residues 50 to 56 of SEQ ID NO:15]

[0511] SEQ ID NO:21—Amino acid sequence of light chain CDRL3 [=amino acid residues 89 to 97 of SEQ ID NO:15]

[0512] SEQ ID NO:22—Amino acid sequence of the heavy chain variable region [=amino acid residues 1 to 120 of SEQ ID NO:14]

[0513] SEQ ID NO:23—Amino acid sequence of light chain variable region [=amino acid residues 1 to 107 of SEQ ID NO:15]

[0514] SEQ ID NO:24—Amino acid sequence of the heavy chain [=amino acid residues 1 to 449 of SEQ ID NO:14]

[0515] SEQ ID NO:25 - Amino acid sequence of anti-PD1 heavy chain CDRH1 of anti-PD-1 / TIGIT bispecific antibody

[0516] SEQ ID NO:26 - Amino acid sequence of anti-PD1 heavy chain CDRH2 of anti-PD-1 / TIGIT bispecific antibody

[0517] SEQ ID NO:27 - Amino acid sequence of anti-PD1 heavy chain CDRH3 of anti-PD-1 / TIGIT bispecific antibody

[0518] SEQ ID NO:28 - Amino acid sequence of anti-PD1 light chain CDRL1 of anti-PD-1 / TIGIT bispecific antibody

[0519] SEQ ID NO:29 - Amino acid sequence of anti-PD1 light chain CDRL2 of anti-PD-1 / TIGIT bispecific antibody

[0520] SEQ ID NO:30 - Amino acid sequence of anti-PD1 light chain CDRL3 of anti-PD-1 / TIGIT bispecific antibody

[0521] SEQ ID NO:31 - Amino acid sequence of the anti-PD1 heavy chain variable region of the anti-PD-1 / TIGIT bispecific antibody

[0522] SEQ ID NO:32 - Amino acid sequence of the anti-PD1 heavy chain of the anti-PD-1 / TIGIT bispecific antibody

[0523] SEQ ID NO:33 - Amino acid sequence of the anti-PD-1 light chain variable region of the anti-PD-1 / TIGIT bispecific antibody

[0524] SEQ ID NO:34 - Amino acid sequence of the anti-PD1 light chain of the anti-PD-1 / TIGIT bispecific antibody

[0525] SEQ ID NO:35 - Amino acid sequence of anti-TIGIT heavy chain CDRH1 of anti-PD-1 / TIGIT bispecific antibody

[0526] SEQ ID NO:36 - Amino acid sequence of anti-TIGIT heavy chain CDRH2 of anti-PD-1 / TIGIT bispecific antibody

[0527] SEQ ID NO:37 - Amino acid sequence of anti-TIGIT heavy chain CDRH3 of anti-PD-1 / TIGIT bispecific antibody

[0528] SEQ ID NO:38 - Amino acid sequence of anti-TIGIT light chain CDRL1 of anti-PD-1 / TIGIT bispecific antibody

[0529] SEQ ID NO:39 - Amino acid sequence of anti-TIGIT light chain CDRL2 of anti-PD-1 / TIGIT bispecific antibody

[0530] SEQ ID NO:40 - Amino acid sequence of anti-TIGIT light chain CDRL3 of anti-PD-1 / TIGIT bispecific antibody

[0531] SEQ ID NO:41 - Amino acid sequence of the anti-TIGIT heavy chain variable region of the anti-PD-1 / TIGIT bispecific antibody

[0532] SEQ ID NO:42 - Amino acid sequence of the anti-TIGIT heavy chain of the anti-PD-1 / TIGIT bispecific antibody

[0533] SEQ ID NO:43 - Amino acid sequence of the anti-TIGIT light chain variable region of the anti-PD-1 / TIGIT bispecific antibody

[0534] SEQ ID NO:44 - Amino acid sequence of the anti-TIGIT light chain of the anti-PD-1 / TIGIT bispecific antibody

[0535] SEQ ID NO:45 - Amino acid sequence of the anti-PD1 light chain of the anti-PD-1 / CTRL-4 bispecific antibody

[0536] SEQ ID NO:46 - Amino acid sequence of the anti-PD1 heavy chain of the anti-PD-1 / CTRL-4 bispecific antibody

[0537] SEQ ID NO:47 - Amino acid sequence of the anti-CTLA-4 light chain of the anti-PD-1 / CTRL-4 bispecific antibody

[0538] SEQ ID NO:48 - Amino acid sequence of the anti-CTLA-4 heavy chain of the anti-PD-1 / CTRL-4 bispecific antibody

[0539] SEQ ID NO:49 - Amino acid sequence of anti-PD1 light chain CDRL1 of anti-PD-1 / CTRL-4 bispecific antibody

[0540] SEQ ID NO:50 - Amino acid sequence of anti-PD1 light chain CDRL2 of anti-PD-1 / CTRL-4 bispecific antibody

[0541] SEQ ID NO:51 - Amino acid sequence of anti-PD1 light chain CDRL3 of anti-PD-1 / CTRL-4 bispecific antibody

[0542] SEQ ID NO:52 - Amino acid sequence of anti-PD1 heavy chain CDRH1 of anti-PD-1 / CTRL-4 bispecific antibody

[0543] SEQ ID NO:53 - Amino acid sequence of anti-PD1 heavy chain CDRH2 of anti-PD-1 / CTRL-4 bispecific antibody

[0544] SEQ ID NO:54 - Amino acid sequence of anti-PD1 heavy chain CDRH3 of anti-PD-1 / CTRL-4 bispecific antibody

[0545] SEQ ID NO:55 - Amino acid sequence of anti-CTLA-4 light chain CDRL1 of anti-PD-1 / CTRL-4 bispecific antibody

[0546] SEQ ID NO:56 - Amino acid sequence of anti-CTLA-4 light chain CDRL2 of anti-PD-1 / CTRL-4 bispecific antibody

[0547] SEQ ID NO:57 - Amino acid sequence of anti-CTLA-4 light chain CDRL3 of anti-PD-1 / CTRL-4 bispecific antibody

[0548] SEQ ID NO:58 - Amino acid sequence of anti-CTLA-4 heavy chain CDRH1 of anti-PD-1 / CTRL-4 bispecific antibody

[0549] SEQ ID NO:59 - Amino acid sequence of anti-CTLA-4 heavy chain CDRH2 of anti-PD-1 / CTRL-4 bispecific antibody

[0550] SEQ ID NO:60—Amino acid sequence of anti-CTLA-4 heavy chain CDRH3 of anti-PD-1 / CTRL-4 bispecific antibody

Claims

1. A pharmaceutical product comprising an antibody-drug conjugate and a bispecific checkpoint inhibitor for combined administration, wherein the antibody-drug conjugate is an antibody-drug conjugate in which a drug-linker represented by the following formula is conjugated to an antibody, preferably an anti-TROP2 or anti-HER2 antibody, via a thioether bond: Wherein A represents the connection position with the antibody.

2. The drug product of claim 1, wherein the drug-linker is conjugated to an anti-TROP2 antibody.

3. The pharmaceutical product according to claim 2, wherein the anti-TROP2 antibody is an antibody comprising a heavy chain and a light chain, the heavy chain comprising 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 the 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.

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

10. 5 . The pharmaceutical product according to claim 4 , wherein 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. 6 . The pharmaceutical product according to claim 4 , wherein 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.

7. The pharmaceutical product according to any one of claims 2 to 6, wherein the average number of drug-linker units conjugated per anti-TROP2 antibody molecule in the antibody-drug conjugate is in the range of 3.5 to 4.

5.

8. The pharmaceutical product according to claim 7, wherein the anti-TROP2 antibody-drug conjugate is deludecant (DS-1062).

9. The drug product of claim 1, wherein the drug-linker is conjugated to an anti-HER2 antibody.

10. The drug product according to claim 9, wherein the anti-HER2 antibody is an antibody comprising a heavy chain and a light chain, the 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 the 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 is an antibody comprising a heavy chain and a light chain, the heavy chain comprising a heavy chain variable region consisting of the amino acid sequence represented by SEQ ID NO: 22, and the 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 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 claim 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 drug product according to any one of claims 9 to 13, wherein the average number of drug-linker units conjugated per anti-HER2 antibody molecule in the antibody-drug conjugate is in the range of 7 to 8.

15. The drug product of claim 14, wherein the anti-HER2 antibody-drug conjugate is trastuzumab (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 pharmaceutical product of claim 16, wherein the bispecific binding protein comprises: a) a first binding domain that specifically binds to PD-1, wherein the first binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDRH1 having an amino acid sequence of SEQ ID NO: 25, CDRH2 having an amino acid sequence of SEQ ID NO: 26, and CDRH3 having an amino acid sequence of SEQ ID NO: 27, and the light chain variable domain comprises CDRL1 having an amino acid sequence of SEQ ID NO: 28, CDRL2 having an amino acid sequence of SEQ ID NO: 29, and CDRL3 having an amino acid sequence of SEQ ID NO: 30; and b) a second binding domain that specifically binds to TIGIT, wherein the second binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDRH1 having an amino acid sequence of SEQ ID NO:35, CDRH2 having an amino acid sequence of SEQ ID NO:36, and CDRH3 having an amino acid sequence of SEQ ID NO:37, and the light chain variable domain comprises CDRL1 having an amino acid sequence of SEQ ID NO:38, CDRL2 having an amino acid sequence of SEQ ID NO:39, and CDRL3 having an amino acid sequence of SEQ ID NO:

40.

18. The drug 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 of SEQ ID NO: 31 and a light chain variable domain having an amino acid sequence of SEQ ID NO:

33.

19. The drug product of 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 an amino acid sequence of SEQ ID NO: 32 and a light chain having an 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 an amino acid sequence of SEQ ID NO:41 and a light chain variable domain having an amino acid sequence of SEQ ID NO:

43.

23. A 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 an amino acid sequence of SEQ ID NO:42 and a light chain having an amino acid sequence of SEQ ID NO:

44.

25. A 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 pharmaceutical product of claim 16, wherein the bispecific binding protein comprises: a) a first binding domain that specifically binds to PD-1, wherein the first binding domain comprises a heavy chain variable domain and a light chain variable domain, the heavy chain variable domain comprises CDRH1 having an amino acid sequence of SEQ ID NO: 52, CDRH2 having an amino acid sequence of SEQ ID NO: 53, and CDRH3 having an amino acid sequence of SEQ ID NO: 54, and the light chain variable domain comprises CDRL1 having an amino acid sequence of SEQ ID NO: 49, CDRL2 having an amino acid sequence of SEQ ID NO: 50, and CDRL3 having an amino acid sequence of SEQ ID NO: 51; and b) a second binding domain that specifically binds to CTLA-4, wherein the second binding domain comprises a heavy chain variable domain and a light chain variable domain, the 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 the 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.

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

45.

28. The drug product of 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 of 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 acid sequence of SEQ ID NO: 48 and a light chain having the amino acid sequence of SEQ ID NO:

47.

30. The drug product of 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 of claim 20 or 27, wherein the light chain constant region is a kappa chain.

32. The pharmaceutical product of 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 drug product of claim 33, wherein the antibody is an IgG antibody.

35. The drug product of claim 34, wherein the antibody is an IgG1 antibody.

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

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 DuetMab.

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

40. The drug product of claim 39, wherein the variant Fc region comprises at least one substitution selected from the group consisting of 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, 2 41E, 241R, 243W, 243L243Y, 243R, 243Q, 244H, 245A, 247L, 247V, 247G, 250E, 250Q, 251F, 252L, 252Y, 254S, 254T, 255L, 256E, 256F, 256M, 257C, 257M, 25 7N, 262I, 262A, 262T, 262E, 263I, 263A, 263T, 263M, 264L, 264I, 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265A, 265G, 265N, 265Q, 265Y, 265F, 265V, 26 5I, 265L, 265H, 265T, 266I, 266A, 266T, 266M, 267Q, 267L, 268E, 269H, 269Y, 269F, 269R, 270E, 280A, 284M, 292P, 292L, 296E, 296Q, 296D, 296N, 296S, 29 6T, 296L, 296I, 296H, 296G, 297S, 297D, 297E, 298A, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 305I, 308F, 313F, 316D, 31 8A, 318S, 320A, 320S, 322A, 322S, 325Q, 325L, 3251, 325D, 325E, 325A, 325T, 325V, 325H, 326A, 326D, 326E, 326G, 326M, 326V, 327G, 327W, 327N, 327L, 32 8S, 328M, 328D, 328E, 328N, 328Q, 328F, 3281, 328V, 328T, 328H, 328A, 329F,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.

41. The pharmaceutical product of claim 39, wherein the variant Fc region comprises one or more amino acid substitutions at positions selected from 428 and 434 numbered by the EU index as set forth in Kabat.

42. The pharmaceutical product according to any one of claims 39 to 41, wherein the variant 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 variant Fc region comprises a 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 aglycosylated Fc region.

45. The pharmaceutical product of 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 with reduced fucosylation or afucosylation.

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

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

49. A pharmaceutical product according to any one of claims 1 to 48, wherein the product is for use in the treatment of cancer.

50. The pharmaceutical product of 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, esophagogastric junction adenocarcinoma, biliary tract cancer, Paget's disease, pancreatic cancer, ovarian cancer, uterine carcinosarcoma, urothelial carcinoma, prostate cancer, bladder cancer, endometrial cancer, gastrointestinal stromal tumors, gastrointestinal stromal tumors, cervical cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, uterine corpus cancer, kidney cancer, vulvar cancer, thyroid cancer, penile cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, glioblastoma multiforme, osteosarcoma, sarcoma, melanoma, cervical cancer, uterine cancer, testicular cancer, and renal cell carcinoma.

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

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

53. The pharmaceutical product of claim 51, wherein the breast cancer is a HER2 low expressing breast cancer.

54. The pharmaceutical product of claim 51 , wherein the breast cancer is triple negative breast cancer.

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

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

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

58. The drug product of claim 57, wherein the non-small cell lung cancer is a non-small cell lung cancer with a targetable genomic alteration.

59. The drug product of claim 57, wherein the non-small cell lung cancer is a non-small cell lung cancer lung cancer without a targetable genomic alteration.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

79. 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 check point inhibitor for treating cancer, wherein the antibody-drug conjugate and the bispecific check point 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 sequentially administering the antibody-drug conjugate and the bispecific check point inhibitor.

83. The antibody-drug conjugate for use according to claim 80 or 81, wherein the use comprises separate and simultaneous administration of the antibody-drug conjugate and the bispecific check point inhibitor.

84. An antibody-drug conjugate for treating cancer in a subject, wherein the treatment comprises administering to the subject i) the antibody-drug conjugate and ii) a bispecific check point inhibitor sequentially or separately simultaneously, wherein the antibody-drug conjugate and the bispecific check point inhibitor are as defined in any one of claims 1 to 46.

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

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

87. A bispecific check point inhibitor for use according to claim 85 or 86, wherein the use comprises sequentially administering the antibody-drug conjugate and the bispecific check point inhibitor.

88. A bispecific check point inhibitor for use according to claim 85 or 86, wherein the use comprises separate and simultaneous administration of the antibody-drug conjugate and the bispecific check point inhibitor.

89. A bispecific check point inhibitor for use in treating cancer in a subject, wherein the treatment comprises administering to the subject sequentially or separately simultaneously i) the bispecific check point inhibitor and ii) an antibody-drug conjugate, wherein the bispecific check point inhibitor and the antibody-drug conjugate are as defined in any one of claims 1 to 46.

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

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

92. The method of claim 90 or 91, wherein the method comprises sequentially administering the antibody-drug conjugate and the bispecific check point inhibitor.

93. The method of claim 90 or 91, wherein the method comprises administering the antibody-drug conjugate and the bispecific check point inhibitor separately and simultaneously.

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