Dosage Regimen of Anti-CDH6 Antibody-Drug Conjugate

Abstract

The present disclosure relates to the fields of pharmaceutical preparations, dosing regimens, and administration of antibody-drug conjugates (ADCs). More specifically, the ADC is composed of an anti-cadherin-6 (CDH6) antibody connected via a linker to an anti-cancer agent such as a topoisomerase I inhibitor.

Description

Technical Field

[0001] Statement Regarding Sequence Listing This application has been electronically filed in XML format and contains a sequence listing that is hereby incorporated by reference in its entirety into this specification. The XML copy created on May 23, 2023, is named 098065-0328.xml and is 118 kb in size.

[0002] This disclosure relates to the field of pharmaceutical preparations, dosage regimens, and administration of antibody-drug conjugates (ADCs). More specifically, the ADC is composed of an anti-cadherin-6 (CDH6) antibody connected via a linker to a topoisomerase I inhibitor such as a derivative of exatecan.

Background Art

[0003] Cadherins are glycoproteins present on the surface of cell membranes and function as cell-cell adhesion molecules or as signaling molecules involved in cell-cell interactions through calcium ion-dependent binding of their N-terminal extracellular domains. Classical cadherins are in the cadherin superfamily and are single-pass transmembrane proteins composed of five extracellular domains (EC domains), one transmembrane region, and an intracellular domain. Classical cadherins are classified into the type I family represented by E-cadherin and N-cadherin, and the type II family, according to the homology of their amino acid sequences.

[0004] Cadherin-6 (CDH6) is a single-pass transmembrane protein composed of 790 amino acids, which is classified into the type II cadherin family, and this protein has an N-terminal extracellular and a C-terminal intracellular domain. The human CDH6 gene was first cloned in 1995 (Non-Patent Document 1), and its sequence can be referred to, for example, under accession numbers NM_004932 and NP_004923 (NCBI).

[0005] CDH6 is specifically expressed in the brain or kidneys at the stage of development and has been reported to play important roles in the formation of central nervous system circuits (Non-Patent Documents 2 and 3) and nephron development in the kidneys (Non-Patent Documents 4 and 5). The expression of CDH6 in normal adult tissues is localized in the renal tubules, bile duct epithelial cells, etc. of the kidneys.

[0006] On the other hand, it is known that CDH6 is specifically overexpressed at the tumor site in some types of adult cancers. The correlation between CDH6 expression and poor prognosis, and its applicability as a tumor marker, have been reported for human renal cell carcinoma, particularly clear cell renal carcinoma (Non-Patent Documents 6 and 7). High expression of CDH6 has also been reported for human ovarian cancer (Non-Patent Document 8). CDH6 has also been reported to be involved in the epithelial-mesenchymal transition of human thyroid cancer (Non-Patent Document 9). Furthermore, CDH6 has been reported to be expressed in human cholangiocarcinoma and human small cell lung cancer as well (Non-Patent Documents 12 and 13).

[0007] Cancer ranks among the top causes of death. Although the number of cancer patients is expected to increase with the aging of the population, the treatment needs have not yet been fully met. The problems with conventional chemotherapeutic drugs are that, due to their low selectivity, these chemotherapeutic drugs are toxic not only to tumor cells but also to normal cells, thereby having adverse reactions; and that the chemotherapeutic drugs cannot be administered in sufficient amounts and thus cannot fully bring about their effects. Therefore, in recent years, more highly selective molecular target drugs or antibody drugs that target molecules that exhibit mutations or high expression characteristics in cancer cells, or specific molecules involved in the malignant transformation of cells, have been developed.

[0008] Antibodies are highly stable in the blood and specifically bind to their target antigens. For these reasons, a reduction in adverse reactions is expected, and numerous antibody drugs have been developed against molecules highly expressed on the surface of cancer cells. One technique that relies on the antigen-specific binding ability of antibodies is the use of antibody-drug conjugates (ADCs). An ADC is a conjugate in which an antibody that can bind to an antigen expressed on the surface of a cancer cell and internalize the antigen into the cell upon binding is conjugated to a drug having cytotoxic activity. It is expected that an ADC can efficiently deliver a drug to cancer cells and thereby kill the cancer cells by accumulating the drug in the cancer cells (Non-Patent Document 10, and Patent Documents 1 and 2). Regarding ADCs, for example, Adcetris (trademark) (brentuximab vedotin), which contains an anti-CD30 monoclonal antibody conjugated to monomethyl auristatin E, is approved as a therapeutic agent for Hodgkin lymphoma and anaplastic large cell lymphoma. Also, Kadcyla (trademark) (trastuzumab emtansine), which contains an anti-HER2 monoclonal antibody conjugated to emtansine, is used in the treatment of HER2-positive advanced or recurrent breast cancer.

[0009] The characteristics of a target antigen suitable for an ADC as an antitumor drug are that the antigen is specifically highly expressed on the surface of cancer cells but has low expression or is not expressed in normal cells; the antigen can be internalized into the cell; the antigen is not secreted from the cell surface; etc. The internalization ability of an antibody depends on the characteristics of both the target antigen and the antibody. It is difficult to predict an antigen-binding site suitable for internalization from the molecular structure of the target or to predict an antibody having a high internalization ability from the binding strength, physical properties, etc. of the antibody. Therefore, an important issue in developing an ADC having high efficacy is to obtain an antibody having a high internalization ability against the target antigen (Non-Patent Document 11).

[0010] An ADC comprising DM4 conjugated to an anti-CDH6 antibody that specifically binds to the EC domain 5 (EC5) of CDH6 is known as an ADC targeting CDH6 (Patent Document 3, Non-Patent Documents 14 and 15). An ADC comprising a derivative of exatecan conjugated to an anti-CDH6 antibody that specifically binds to the EC domain 3 (EC3) of CDH6 is known (Patent Documents 4 and 5).

Prior Art Documents

Patent Documents

[0011]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Non-Patent Documents

[0012]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Non-Patent Document 7

Non-Patent Document 8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 13

Non-Patent Document 14

Non-Patent Document 15

Summary of the Invention

[0013] The present disclosure provides a method for treating cancer in a subject in need thereof, comprising administering a specific anti-CDH6 antibody-drug conjugate to the subject according to a specific dosage regimen, and a pharmaceutical composition for use in treating cancer according to the dosage regimen. The present disclosure also provides a method for treating renal cell carcinoma or ovarian cancer in a subject in need thereof, comprising administering a specific anti-CDH6 antibody-drug conjugate to the subject, and a pharmaceutical composition for use in treating renal cell carcinoma or ovarian cancer, comprising the anti-CDH6 antibody-drug conjugate.

[0014] The present disclosure includes the following aspects: [1] An anti-CDH6 antibody-drug conjugate for use in treating or preventing cancer, wherein the antibody-drug conjugate comprises an anti-CDH6 antibody and an anti-tumor compound connected by a linker, and the linker and the anti-tumor compound have the following formula: -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-(NH-DX) as represented by wherein -(Succinimid-3-yl-N)- has the structure represented by the following formula:

[0015]

Chemical formula

[0016]

Chemical formula

[0017] [Chemical formula] has a structure represented by, wherein Ab is an antibody comprising a heavy chain variable region containing SEQ ID NO: 71 and a light chain variable region containing SEQ ID NO: 63, n represents the average number of units of the drug-linker structure conjugated to the antibody per antibody, and the average number of units of the conjugated selected drug-linker structure per antibody is in the range of 1 to 10, the antibody-drug conjugate according to [1] or [2]. [4] The antibody comprises a heavy chain containing the amino acid sequence at positions 20 to 471 in SEQ ID NO: 69 and a light chain containing the amino acid sequence at positions 21 to 233 in SEQ ID NO: 61, the antibody-drug conjugate according to any one of [1] to [3]. [5] The heavy or light chain is subjected to one or more types of modifications selected from the group consisting of N-linked glycosylation, O-linked glycosylation, N-terminal processing, C-terminal processing, deamidation, isomerization of aspartic acid, oxidation of methionine, addition of a methionine residue to the N-terminus, amidation of a proline residue, conversion of an N-terminal glutamine or N-terminal glutamic acid to pyroglutamic acid, and deletion of one or two amino acids from the carboxyl terminus, the antibody-drug conjugate according to any one of [1] to [4]. [6] The anti-CDH6 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain, the antibody-drug conjugate according to any one of [1] to [5]. [7] The average number of units of the conjugated anti-tumor compound per antibody is in the range of 2 to 8, the antibody-drug conjugate according to any one of [1] to [6]. [8] The average number of units of the conjugated anti-tumor compound per antibody is 7 to 8, the antibody-drug conjugate according to any one of [1] to [7]. [9] A dose of the antibody-drug conjugate in the range of 1.6 mg / kg to 9.6 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[10] A dose of the antibody-drug conjugate in the range of 3.2 mg / kg to 9.6 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[11] A dose of the antibody-drug conjugate in the range of 4.8 mg / kg to 9.6 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[12] A dose of the antibody-drug conjugate in the range of 3.2 mg / kg to 8.0 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[13] An antibody-drug conjugate in the range of 3.2 mg / kg to 6.4 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[14] An antibody-drug conjugate in the range of 4.8 mg / kg to 8.0 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[15] An antibody-drug conjugate in the range of 4.8 mg / kg to 6.4 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[16] A dose of the antibody-drug conjugate of about 3.2 mg / kg, about 4.8 mg / kg, about 5.4 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg, or about 8.0 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[17] A dose of the antibody-drug conjugate of about 3.2 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[18] A dose of the antibody-drug conjugate of about 4.8 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[19] A dose of the antibody-drug conjugate of about 5.4 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[20] A dose of the antibody-drug conjugate of about 5.6 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[21] A dose of the antibody-drug conjugate of about 6.4 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[22] A dose of the antibody-drug conjugate of about 8.0 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[23] A dosage of the antibody-drug conjugate of 3.2 mg / kg, 3.3 mg / kg, 3.4 mg / kg, 3.5 mg / kg, 3.6 mg / kg, 3.7 mg / kg, 3.8 mg / kg, 3.9 mg / kg, 4.0 mg / kg, 4.1 mg / kg, 4.2 mg / kg, 4.3 mg / kg, 4.4 mg / kg, 4.5 mg / kg, 4.6 mg / kg, 4.7 mg / kg, 4.8 mg / kg, 4.9 mg / kg, 5.0 mg / kg, 5.1 mg / kg, 5.2 mg / kg, 5.3 mg / kg, 5.4 mg / kg, 5.5 mg / kg, 5.6 mg / kg, 5.7 mg / kg, 5.8 mg / kg, 5.9 mg / kg, 6.0 mg / kg, 6.1 mg / kg, 6.2 mg / kg, 6.3 mg / kg, or 6.4 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1] to [8].

[24] The antibody-drug conjugate according to any one of [1] to

[23] , which is administered by intravenous administration.

[25] The antibody-drug conjugate according to any one of [1] to

[24] , which is administered once every three weeks.

[26] The cancer is selected from the group consisting of renal cell carcinoma, clear cell renal carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, clear cell carcinoma of the ovary, endometrioid carcinoma of the ovary, mucinous tumor of the ovary, thyroid cancer, cholangiocarcinoma, lung cancer, non-small cell lung cancer, cervical cancer, brain tumor, head and neck cancer, sarcoma, osteosarcoma, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, neuroblastoma, colorectal cancer, gastric cancer, endometrial cancer, nasopharyngeal cancer, prostate cancer, or cancer associated with von Hippel-Lindau disease, the antibody-drug conjugate according to any one of [1] to

[25] .

[27] The cancer is renal cell carcinoma, clear cell renal carcinoma, or papillary renal cell carcinoma, the antibody-drug conjugate according to

[26] .

[28] The cancer is ovarian cancer, the antibody-drug conjugate according to

[26] .

[29] The ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal cancer, the antibody-drug conjugate according to

[28] . The antibody-drug conjugate according to any one of [1] to

[29] , wherein the cancer is metastatic. The antibody-drug conjugate according to any one of [1] to

[30] , wherein the cancer is resistant or refractory. The antibody-drug conjugate according to

[31] , wherein the resistance or refractoriness is resistance or refractoriness acquired by the cancer due to treatment with an anticancer drug. The antibody-drug conjugate according to

[32] , wherein the anticancer drug is a platinum-based chemotherapeutic agent, a chemotherapeutic agent, a PARP inhibitor, an immune checkpoint inhibitor, an angiogenesis inhibitor, or a VEGFR-TKI. The antibody-drug conjugate according to any one of [1] to

[33] , wherein the subject has a treatment history with one or more types of anticancer drugs selected from the group consisting of a platinum-based chemotherapeutic agent, a chemotherapeutic agent, a PARP inhibitor, an immune checkpoint inhibitor, an angiogenesis inhibitor, and a VEGFR-TKI. The antibody-drug conjugate according to

[32] , wherein the anticancer drug is a platinum-based chemotherapeutic agent. The antibody-drug conjugate according to

[35] , wherein the platinum-based chemotherapeutic agent includes a platinum-based drug and a taxane. The antibody-drug conjugate according to any one of [1] to

[36] , wherein the cancer is a CDH6-expressing cancer. The antibody-drug conjugate according to

[37] , wherein the CDH6-expressing cancer is a CDH6-overexpressing cancer. The antibody-drug conjugate according to any one of [1] to

[38] , wherein the cancer is inoperable or recurrent cancer. The antibody-drug conjugate according to any one of [1] to

[39] , wherein the subject exhibits recurrence of cancer before administration of the antibody-drug conjugate. The antibody-drug conjugate according to

[40] , wherein the recurrence of cancer occurs within or less than about 6 months after completion of a chemotherapy regimen including a platinum-based drug.

[42] Cancer recurrence is the antibody-drug conjugate described in

[40] that occurs less than about 6 months after or within about 6 months after completion of a chemotherapy regimen including platinum-based drugs and taxanes.

[43] Cancer recurrence is the antibody-drug conjugate described in

[40] that occurs about 6 months after or after about 6 months have elapsed after completion of a chemotherapy regimen including platinum-based drugs.

[44] Cancer recurrence is the antibody-drug conjugate described in

[40] that occurs about 6 months after or after about 6 months have elapsed after completion of a chemotherapy regimen including platinum-based drugs and taxanes.

[45] The objective response rate of the subject is at least about 20%, and the antibody-drug conjugate described in any one of [1] to

[44] .

[46] The objective response rate of the subject is at least about 30%, and the antibody-drug conjugate described in any one of [1] to

[44] .

[47] The progression-free survival period of the subject is at least 5 months after administration of the antibody-drug conjugate, and the antibody-drug conjugate described in any one of [1] to

[46] .

[48] The progression-free survival period of the subject is at least 5.5 months after administration of the antibody-drug conjugate, and the antibody-drug conjugate described in any one of [1] to

[46] .

[49] The antibody-drug conjugate described in any one of [1] to

[48] , which is administered as a single-agent therapy.

[50] The subject is administered an antibody-drug conjugate together with a second drug, and the antibody-drug conjugate described in any one of [1] to

[48] .

[51] The antibody-drug conjugate described in

[50] , which is administered before, after, or simultaneously with the second drug.

[52] The antibody-drug conjugate described in any one of [1] to

[44] , which is administered as maintenance therapy.

[53] The antibody-drug conjugate described in any one of [1] to

[44] , which is administered as adjuvant therapy.

[54] An antibody-drug conjugate according to any one of [1] to

[44] , which is administered after surgical resection of a tumor.

[55] An antibody-drug conjugate according to any one of [1] to

[44] , which is administered as neoadjuvant therapy.

[56] An antibody-drug conjugate according to any one of [1] to

[44] , which is administered before surgical resection of a tumor.

[57] The following formula:

[0018] [Chemical formula] An anti-CDH6 antibody-drug conjugate for use in treating or preventing cancer, having a structure represented by: In the formula, AB represents an anti-CDH6 antibody or a functional fragment of the antibody, n represents the average number of units of the drug-linker structure conjugated to the antibody per antibody, and the antibody is connected to the linker via a sulfhydryl group derived from the antibody; the anti-CDH6 antibody-drug conjugate is a salt thereof, or a hydrate or salt of the anti-CDH6 antibody-drug conjugate, the average number of units of the conjugated drug-linker structure per antibody is 7 to 8, the antibody comprises a heavy chain amino acid sequence represented by SEQ ID NO: 87, or an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 87 with one or two amino acids deleted from its carboxyl terminus, and a light chain amino acid sequence represented by SEQ ID NO: 88; the dosage of the antibody-drug conjugate is in the range of 1.6 mg / kg to 9.6 mg / kg, the antibody-drug conjugate is administered by intravenous administration, and the antibody-drug conjugate is administered once every three weeks. An anti-CDH6 antibody-drug conjugate.

[58] A pharmaceutical composition containing an antibody-drug conjugate according to any one of [1] to

[57] or a salt thereof as an active ingredient, and a pharmaceutically acceptable formulation ingredient. A method of treating or preventing cancer in a subject, comprising administering to the subject having cancer an anti-CDH6 antibody-drug conjugate comprising an anti-CDH6 antibody and an anti-tumor compound connected by a linker, wherein the linker and the anti-tumor compound have the following formula: -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-(NH-DX) represented by wherein -(Succinimid-3-yl-N)- has the structure represented by the following formula:

[0019]

Chemical formula

[0020]

Chemical formula

[60] The method according to

[59] , wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 71 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 63.

[61] The antibody-drug conjugate has the following formula:

[0021] [Chemical formula] has a structure represented by wherein Ab is an antibody comprising a heavy chain variable region containing SEQ ID NO: 71 and a light chain variable region containing SEQ ID NO: 63, and n represents the average number of units of the drug-linker structure conjugated to the antibody per antibody, and the average number of units of the conjugated selected drug-linker structure per antibody is in the range of 1 to 10, the method described in

[59] or

[60] .

[62] The antibody comprises a heavy chain containing the amino acid sequence at positions 20 to 471 in SEQ ID NO: 69 and a light chain containing the amino acid sequence at positions 21 to 233 in SEQ ID NO: 61, the method described in any one of

[59] to

[61] .

[63] The heavy chain or light chain has undergone one or more types of modifications selected from the group consisting of N-linked glycosylation, O-linked glycosylation, N-terminal processing, C-terminal processing, deamidation, asparagine isomerization, methionine oxidation, addition of a methionine residue to the N-terminus, amidation of a proline residue, conversion of the N-terminal glutamine or N-terminal glutamic acid to pyroglutamic acid, and deletion of one or two amino acids from the carboxyl terminus, the method described in any one of

[59] to

[62] .

[64] The anti-CDH6 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain, the method described in any one of

[59] to

[63] .

[65] The average number of units of the conjugated anti-tumor compound per antibody is in the range of 2 to 8, the method described in any one of

[59] to

[64] .

[66] The average number of units of the conjugated anti-tumor compound per antibody is 7 to 8, the method described in any one of

[59] to

[65] .

[67] A dose of the antibody-drug conjugate in the range of 1.6 mg / kg to 9.6 mg / kg is administered to a subject having cancer, the method described in any one of

[59] to

[66] .

[68] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate within the range of 3.2 mg / kg to 9.6 mg / kg is administered to a subject having cancer.

[69] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate within the range of 4.8 mg / kg to 9.6 mg / kg is administered to a subject having cancer.

[70] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate within the range of 3.2 mg / kg to 8.0 mg / kg is administered to a subject having cancer.

[71] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate within the range of 3.2 mg / kg to 6.4 mg / kg is administered to a subject having cancer.

[72] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate within the range of 4.8 mg / kg to 8.0 mg / kg is administered to a subject having cancer.

[73] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate within the range of 4.8 mg / kg to 6.4 mg / kg is administered to a subject having cancer.

[74] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate of about 3.2 mg / kg, about 4.8 mg / kg, about 5.4 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg, or about 8.0 mg / kg is administered to a subject having cancer.

[75] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate of about 3.2 mg / kg is administered to a subject having cancer.

[76] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate of about 4.8 mg / kg is administered to a subject having cancer.

[77] The method according to any one of

[59] to

[66] , wherein an amount of the antibody-drug conjugate of about 5.4 mg / kg is administered to a subject having cancer.

[78] The method according to any one of

[59] to

[66] , wherein a dose of the antibody-drug conjugate of about 5.6 mg / kg is administered to a subject having cancer.

[79] The method according to any one of

[59] to

[66] , wherein a dose of the antibody-drug conjugate of about 6.4 mg / kg is administered to a subject having cancer.

[80] The method according to any one of

[59] to

[66] , wherein a dose of the antibody-drug conjugate of about 8.0 mg / kg is administered to a subject having cancer.

[81] The method according to any one of

[59] to

[66] , wherein a dose of the antibody-drug conjugate of 3.2 mg / kg, 3.3 mg / kg, 3.4 mg / kg, 3.5 mg / kg, 3.6 mg / kg, 3.7 mg / kg, 3.8 mg / kg, 3.9 mg / kg, 4.0 mg / kg, 4.1 mg / kg, 4.2 mg / kg, 4.3 mg / kg, 4.4 mg / kg, 4.5 mg / kg, 4.6 mg / kg, 4.7 mg / kg, 4.8 mg / kg, 4.9 mg / kg, 5.0 mg / kg, 5.1 mg / kg, 5.2 mg / kg, 5.3 mg / kg, 5.4 mg / kg, 5.5 mg / kg, 5.6 mg / kg, 5.7 mg / kg, 5.8 mg / kg, 5.9 mg / kg, 6.0 mg / kg, 6.1 mg / kg, 6.2 mg / kg, 6.3 mg / kg, or 6.4 mg / kg is administered to a subject having cancer.

[82] The method according to any one of

[59] or

[81] , wherein the antibody-drug conjugate is administered by intravenous administration.

[83] The method according to any one of

[59] to

[82] , wherein the antibody-drug conjugate is administered once every three weeks.

[84] The cancer is selected from the group consisting of renal cell carcinoma, clear cell renal carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, clear cell carcinoma of the ovary, endometrioid carcinoma of the ovary, ovarian mucinous tumor, thyroid cancer, cholangiocarcinoma, lung cancer, non-small cell lung cancer, cervical cancer, brain tumor, head and neck cancer, sarcoma, osteosarcoma, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, neuroblastoma, colorectal cancer, gastric cancer, endometrial cancer, nasopharyngeal cancer, prostate cancer, or cancer associated with von Hippel-Lindau disease, according to any one of

[59] to

[83] .

[85] The method according to

[84] , wherein the cancer is renal cell carcinoma, clear cell renal cell carcinoma, or papillary renal cell carcinoma.

[86] The method according to

[84] , wherein the cancer is ovarian cancer.

[87] The method according to

[86] , wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal cancer.

[88] The antibody-drug conjugate according to any one of

[59] to

[87] , wherein the cancer is metastatic.

[89] The method according to any one of

[59] to

[88] , wherein the cancer is resistant or refractory.

[90] The method according to

[89] , wherein the resistance or refractoriness is resistance or refractoriness obtained by the cancer due to treatment with an anticancer drug.

[91] The method according to

[90] , wherein the anticancer drug is a platinum-based chemotherapeutic agent, chemotherapeutic agent, PARP inhibitor, immune checkpoint inhibitor, angiogenesis inhibitor, or VEGFR-TKI.

[92] The method according to any one of

[59] to

[91] , wherein the subject has a treatment history with one or more types of anticancer drugs selected from the group consisting of a platinum-based chemotherapeutic agent, chemotherapeutic agent, PARP inhibitor, immune checkpoint inhibitor, angiogenesis inhibitor, and VEGFR-TKI.

[93] The method according to

[90] , wherein the anticancer drug is a platinum-based chemotherapeutic agent.

[94] The method according to

[93] , wherein the platinum-based chemotherapeutic agent includes a platinum-based drug and taxane.

[95] The method according to any one of

[59] to

[94] , wherein the cancer is CDH6-expressing cancer.

[96] The method according to

[95] , wherein the CDH6-expressing cancer is CDH6-overexpressing cancer.

[97] The method according to any one of

[59] to

[96] , wherein the cancer is inoperable or recurrent cancer.

[98] The method according to any one of

[59] to

[97] , wherein the subject exhibits recurrence of cancer before administration of the antibody-drug conjugate. The recurrence of cancer is the method described in

[98] that occurs less than or within about 6 months after completion of a chemotherapy regimen including a platinum-based drug.

[0100] The recurrence of cancer is the method described in

[98] that occurs less than or within about 6 months after completion of a chemotherapy regimen including a platinum-based drug and a taxane.

[0101] The recurrence of cancer is the method described in

[98] that occurs about 6 months after or after about 6 months have elapsed since completion of a chemotherapy regimen including a platinum-based drug.

[0102] The recurrence of cancer is the method described in

[98] that occurs about 6 months after or after about 6 months have elapsed since completion of a chemotherapy regimen including a platinum-based drug and a taxane.

[0103] The objective response rate of the subject is at least about 20%, the method described in any one of

[59] to

[0102] .

[0104] The objective response rate of the subject is at least about 30%, the method described in any one of

[59] to

[0102] .

[0105] The progression-free survival period of the subject is at least 5 months after administration of the antibody-drug conjugate, the method described in any one of

[59] to

[0104] .

[0106] The progression-free survival period of the subject is at least 5.5 months after administration of the antibody-drug conjugate, the method described in any one of

[59] to

[0104] .

[0107] The antibody-drug conjugate is administered as a monotherapy, the method described in any one of

[59] to

[0106] .

[0108] The subject is administered the antibody-drug conjugate together with a second drug, the method described in any one of

[59] to

[0106] .

[0109] The antibody-drug conjugate is administered before, after, or simultaneously with the second drug, the method described in

[0108] .

[0110] The antibody-drug conjugate is administered as maintenance therapy, the method described in any one of

[59] to

[0102] .

[0111] The method according to any one of

[59] to

[0102] , wherein the antibody-drug conjugate is administered as adjuvant therapy.

[0112] The method according to any one of

[59] to

[0102] , wherein the antibody-drug conjugate is administered after surgical resection of the tumor.

[0113] The method according to any one of

[59] to

[0102] , wherein the antibody-drug conjugate is administered as neoadjuvant therapy.

[0114] The method according to any one of

[59] to

[0102] , wherein the antibody-drug conjugate is administered before surgical resection of the tumor.

[0115] The following formula:

[0022] [Chemical formula] A method for treating or preventing cancer in a subject, comprising the step of administering to the subject having cancer an anti-CDH6 antibody-drug conjugate having a structure represented by: In the formula, AB represents an anti-CDH6 antibody or a functional fragment of the antibody, n represents the average number of units of the drug-linker structure conjugated to the antibody per antibody, and the antibody is connected to the linker via a sulfhydryl group derived from the antibody; the anti-CDH6 antibody-drug conjugate is a salt thereof, or a hydrate or salt of the anti-CDH6 antibody-drug conjugate, and the average number of units of the conjugated drug-linker structure per antibody is 7 to 8, and the antibody comprises a heavy chain amino acid sequence represented by SEQ ID NO: 87, or an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 87 in which one or two amino acids are deleted from its carboxyl terminus; and a light chain amino acid sequence represented by SEQ ID NO: 88, and the dose of the antibody-drug conjugate is in the range of 1.6 mg / kg to 9.6 mg / kg, and the antibody-drug conjugate is administered by intravenous administration, and the antibody-drug conjugate is administered once every three weeks.

[0116] The method according to any one of

[59] to

[0115] , wherein the antibody-drug conjugate is administered in the form of a pharmaceutical composition comprising at least one pharmaceutically acceptable formulation ingredient.

[0117] Use of an anti-CDH6 antibody-drug conjugate in the manufacture of a medicament for treating or preventing cancer, wherein the antibody-drug conjugate comprises an anti-CDH6 antibody and an anti-tumor compound connected by a linker, and the linker and the anti-tumor compound have the following formula: -(Succinimid-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GGFG-NH-CH2-O-CH2-C(=O)-(NH-DX) represented by wherein, -(Succinimid-3-yl-N)- has the following formula:

[0023]

Chemical formula

[0024]

Chemical formula

[0118] The use according to

[0117] , wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 71 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 63.

[0119] An antibody-drug conjugate has the structure represented by the following formula:

[0025] [Chemical formula] and has the structure represented by wherein Ab is an antibody comprising a heavy chain variable region containing SEQ ID NO: 71 and a light chain variable region containing SEQ ID NO: 63; n represents the average number of units of the drug-linker structure conjugated to the antibody per antibody, and the average number of units of the conjugated selected drug-linker structure per antibody is in the range of 1 to 10, the use according to

[0117] or

[0118] .

[0120] The antibody comprises a heavy chain containing the amino acid sequence at positions 20 to 471 in SEQ ID NO: 69 and a light chain containing the amino acid sequence at positions 21 to 233 in SEQ ID NO: 61, the use according to any one of

[0117] to

[0119] .

[0121] The heavy chain or the light chain has undergone one or more types of modifications selected from the group consisting of N-linked glycosylation, O-linked glycosylation, N-terminal processing, C-terminal processing, deamidation, asparagine isomerization, methionine oxidation, addition of a methionine residue to the N-terminus, amidation of a proline residue, conversion of the N-terminal glutamine or N-terminal glutamic acid to pyroglutamic acid, and deletion of one or two amino acids from the carboxyl terminus, the use according to any one of

[0117] to

[0120] .

[0122] The anti-CDH6 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain, the use according to any one of

[0117] to

[0121] .

[0123] The average number of units of the conjugated anti-tumor compound per antibody is in the range of 2 to 8, the use according to any one of

[0117] to

[0122] .

[0124] The average number of units of the conjugated anti-tumor compound per antibody is 7 to 8, the use according to any one of

[0117] to

[0123] .

[0125] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate within the range of 1.6 mg / kg to 9.6 mg / kg is administered to a subject having cancer.

[0126] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate within the range of 3.2 mg / kg to 9.6 mg / kg is administered to a subject having cancer.

[0127] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate within the range of 4.8 mg / kg to 9.6 mg / kg is administered to a subject having cancer.

[0128] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate within the range of 3.2 mg / kg to 8.0 mg / kg is administered to a subject having cancer.

[0129] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate within the range of 3.2 mg / kg to 6.4 mg / kg is administered to a subject having cancer.

[0130] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate within the range of 4.8 mg / kg to 8.0 mg / kg is administered to a subject having cancer.

[0131] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate within the range of 4.8 mg / kg to 6.4 mg / kg is administered to a subject having cancer.

[0132] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate of about 3.2 mg / kg, about 4.8 mg / kg, about 5.4 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg, or about 8.0 mg / kg is administered to a subject having cancer.

[0133] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate of about 3.2 mg / kg is administered to a subject having cancer.

[0134] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate of about 4.8 mg / kg is administered to a subject having cancer.

[0135] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate of about 5.4 mg / kg is administered to a subject having cancer.

[0136] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate of about 5.6 mg / kg is administered to a subject having cancer.

[0137] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate of about 6.4 mg / kg is administered to a subject having cancer.

[0138] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate of about 8.0 mg / kg is administered to a subject having cancer.

[0139] The use according to any one of

[0117] to

[0124] , wherein a dose of the antibody-drug conjugate of 3.2 mg / kg, 3.3 mg / kg, 3.4 mg / kg, 3.5 mg / kg, 3.6 mg / kg, 3.7 mg / kg, 3.8 mg / kg, 3.9 mg / kg, 4.0 mg / kg, 4.1 mg / kg, 4.2 mg / kg, 4.3 mg / kg, 4.4 mg / kg, 4.5 mg / kg, 4.6 mg / kg, 4.7 mg / kg, 4.8 mg / kg, 4.9 mg / kg, 5.0 mg / kg, 5.1 mg / kg, 5.2 mg / kg, 5.3 mg / kg, 5.4 mg / kg, 5.5 mg / kg, 5.6 mg / kg, 5.7 mg / kg, 5.8 mg / kg, 5.9 mg / kg, 6.0 mg / kg, 6.1 mg / kg, 6.2 mg / kg, 6.3 mg / kg, or 6.4 mg / kg is administered to a subject having cancer.

[0140] The use according to any one of

[0117] to

[0139] , wherein the antibody-drug conjugate is administered by intravenous administration.

[0141] The use according to any one of

[0117] to

[0140] , wherein the antibody-drug conjugate is administered once every three weeks.

[0142] The use according to any one of

[0117] to

[0141] , wherein the cancer is selected from the group consisting of renal cell carcinoma, clear cell renal carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, clear cell carcinoma of the ovary, endometrioid carcinoma of the ovary, mucinous tumor of the ovary, thyroid cancer, cholangiocarcinoma, lung cancer, non-small cell lung cancer, cervical cancer, brain tumor, head and neck cancer, sarcoma, osteosarcoma, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, neuroblastoma, colorectal cancer, gastric cancer, endometrial cancer, nasopharyngeal cancer, prostate cancer, or cancer associated with von Hippel-Lindau disease.

[0143] The use according to

[0142] , wherein the cancer is renal cell carcinoma, clear cell renal carcinoma, or papillary renal cell carcinoma.

[0144] The use according to

[0142] , wherein the cancer is ovarian cancer.

[0145] The use according to

[0144] , wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal cancer.

[0146] The use according to any one of

[0117] to

[0145] , wherein the cancer is metastatic.

[0147] The use according to any one of

[0117] to

[0146] , wherein the cancer is resistant or refractory.

[0148] The use according to

[0147] , wherein the resistance or refractoriness is resistance or refractoriness obtained by the cancer due to treatment with an anticancer drug.

[0149] The use according to

[0148] , wherein the anticancer drug is a platinum-based chemotherapeutic agent, chemotherapeutic agent, PARP inhibitor, immune checkpoint inhibitor, angiogenesis inhibitor, or VEGFR-TKI.

[0150] The subject has a treatment history using one or more types of anticancer drugs selected from the group consisting of platinum-based chemotherapeutic drugs, chemotherapeutic drugs, PARP inhibitors, immune checkpoint inhibitors, angiogenesis inhibitors, and VEGFR-TKIs, the use according to any one of

[0117] to

[0149] .

[0151] The anticancer drug is a platinum-based chemotherapeutic drug, the use according to

[0148] .

[0152] The platinum-based chemotherapeutic drug includes platinum-based drugs and taxanes, the use according to

[0152] .

[0153] The cancer is CDH6-expressing cancer, the use according to any one of

[0117] to

[0152] .

[0154] The CDH6-expressing cancer is CDH6-overexpressing cancer, the use according to

[0153] .

[0155] The cancer is inoperable or recurrent cancer, the use according to any one of

[0117] to

[0154] .

[0156] The subject exhibits cancer recurrence before the administration of the antibody-drug conjugate, the use according to any one of

[0117] to

[0155] .

[0157] The cancer recurrence occurs less than about 6 months after or within the completion of a chemotherapy regimen including a platinum-based drug, the use according to

[0156] .

[0158] The cancer recurrence occurs less than about 6 months after or within the completion of a chemotherapy regimen including a platinum-based drug and taxanes, the use according to

[0156] .

[0159] The cancer recurrence occurs about 6 months after or after about 6 months have elapsed since the completion of a chemotherapy regimen including a platinum-based drug, the use according to

[0156] .

[0160] The cancer recurrence occurs about 6 months after or after about 6 months have elapsed since the completion of a chemotherapy regimen including a platinum-based drug and taxanes, the use according to

[0156] .

[0161] The objective efficacy rate of the subject is at least about 20%, the use according to any one of

[0117] to

[0160] .

[0162] The objective efficacy rate of the subject is at least about 30%, the use according to any one of

[0117] to

[0160] .

[0163] The progression-free survival period of the subject is at least 5 months after administration of the antibody-drug conjugate, the use according to any one of

[0117] to

[0162] .

[0164] The progression-free survival period of the subject is at least 5.5 months after administration of the antibody-drug conjugate, the use according to any one of

[0117] to

[0162] .

[0165] The antibody-drug conjugate is administered as a monotherapy, the use according to any one of

[0117] to

[0164] .

[0166] The subject is administered the antibody-drug conjugate together with a second drug, the use according to any one of

[0117] to

[0164] .

[0167] The antibody-drug conjugate is administered before, after, or simultaneously with the second drug, the use according to

[0166] .

[0168] The antibody-drug conjugate is administered as maintenance therapy, the use according to any one of

[0117] to

[0160] .

[0169] The antibody-drug conjugate is administered as adjuvant therapy, the use according to any one of

[0117] to

[0160] .

[0170] The antibody-drug conjugate is administered after surgical resection of the tumor, the use according to any one of

[0117] to

[0160] .

[0171] The antibody-drug conjugate is administered as neoadjuvant therapy, the use according to any one of

[0117] to

[0160] .

[0172] The antibody-drug conjugate is administered before surgical resection of the tumor, the use according to any one of

[0117] to

[0160] .

[0173] Use of an anti-CDH6 antibody-drug conjugate in the manufacture of a medicament for treating or preventing cancer, wherein the anti-CDH6 antibody-drug conjugate has the following formula:

[0026] [Chemical formula] having a structure represented by: In the formula, AB represents an anti-CDH6 antibody or a functional fragment of the antibody, n represents the average number of units of the drug-linker structure conjugated to the antibody per antibody, and the antibody is connected to the linker via a sulfhydryl group derived from the antibody; the anti-CDH6 antibody-drug conjugate is a salt thereof, or a hydrate or salt of the anti-CDH6 antibody-drug conjugate, and the average number of units of the conjugated drug-linker structure per antibody is 7 to 8, and the antibody comprises a heavy chain amino acid sequence represented by SEQ ID NO: 87, or an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 87 with one or two amino acids deleted from its carboxyl terminus; and a light chain amino acid sequence represented by SEQ ID NO: 88, and the dose of the antibody-drug conjugate is in the range of 1.6 mg / kg to 9.6 mg / kg, and the antibody-drug conjugate is administered by intravenous administration, and the antibody-drug conjugate is administered once every three weeks.

[0174] The use according to any one of

[0117] to

[0173] , wherein the antibody-drug conjugate is administered in the form of a pharmaceutical composition comprising at least one pharmaceutically acceptable formulation ingredient.

[0027] The present invention also includes the following aspects of the present invention: [1A] An anti-CDH6 antibody-drug conjugate for use in treating or preventing cancer, wherein the antibody-drug conjugate comprises an anti-CDH6 antibody and an antitumor compound connected by a linker, and the linker and the antitumor compound have the following formula: -(Succinimid-3-yl-N)-CH 2 CH 2 CH2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-(NH-DX) represented by wherein, -(Succinimid-3-yl-N)- is the following formula:

[0028] [Chemical formula] has a structure represented by, is connected to the antibody at the 3-position, and is connected to the methylene group in the linker structure containing this structure on the nitrogen atom at the 1-position. (NH-DX) is the following formula:

[0029] [Chemical formula] represents a group represented by, the nitrogen atom of the amino group at the 1-position is the connection position, and the anti-CDH6 antibody comprises CDRH1 consisting of the amino acid sequence of SEQ ID NO: 17, CDRH2 consisting of the amino acid sequence of SEQ ID NO: 60, and CDRH3 consisting of the amino acid sequence of SEQ ID NO: 19 in its heavy chain variable region, and CDRL1 consisting of the amino acid sequence of SEQ ID NO: 12, CDRL2 consisting of the amino acid sequence of SEQ ID NO: 13, and CDRL3 consisting of the amino acid sequence of SEQ ID NO: 14 in its light chain variable region, an anti-CDH6 antibody-drug conjugate. [2A] The antibody comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 71 and a light chain variable region containing the amino acid sequence of SEQ ID NO: 63, the antibody-drug conjugate according to [1A]. [3A] The following formula:

[0030] [Chemical formula] has a structure represented by In the formula, Ab is an antibody comprising a heavy chain variable region containing SEQ ID NO: 71 and a light chain variable region containing SEQ ID NO: 63, n represents the average number of units of the drug-linker structure conjugated to the antibody per antibody, and the average number of units of the conjugated selected drug-linker structure per antibody is within the range of 1 to 10, the antibody-drug conjugate described in [1A] or [2A]. [4A] The antibody comprises a heavy chain containing the amino acid sequence at positions 20 to 471 in SEQ ID NO: 69 and a light chain containing the amino acid sequence at positions 21 to 233 in SEQ ID NO: 61, the antibody-drug conjugate according to any one of [1A] to [3A]. [5A] The heavy chain or the light chain has undergone one or more types of modifications selected from the group consisting of N-linked glycosylation, O-linked glycosylation, N-terminal processing, C-terminal processing, deamidation, asparagine isomerization, methionine oxidation, addition of a methionine residue to the N-terminus, amidation of a proline residue, conversion of the N-terminal glutamine or N-terminal glutamate to pyroglutamic acid, and deletion of one or two amino acids from the carboxyl terminus, the antibody-drug conjugate according to any one of [1A] to [4A]. [6A] The anti-CDH6 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain, the antibody-drug conjugate according to any one of [1A] to [5A]. [7A] The average number of units of the conjugated antitumor compound per antibody is within the range of 2 to 8, the antibody-drug conjugate according to any one of [1A] to [6A]. [8A] The average number of units of the conjugated antitumor compound per antibody is 7 to 8, the antibody-drug conjugate according to any one of [1A] to [7A]. [9A] A dose of the antibody-drug conjugate within the range of 1.6 mg / kg to 9.6 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1A] to [8A]. [10A] An antibody-drug conjugate with a dose of about 3.2 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1A] to [9A]. [11A] An antibody-drug conjugate with a dose of about 4.8 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1A] to [9A]. [12A] An antibody-drug conjugate with a dose of about 6.4 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1A] to [9A]. [13A] An antibody-drug conjugate with a dose of about 8.0 mg / kg is administered to a subject having cancer, the antibody-drug conjugate according to any one of [1A] to [9A]. [14A] The antibody-drug conjugate according to any one of [1A] to [13A], which is administered by intravenous administration. [15A] The antibody-drug conjugate according to any one of [1A] to [14A], which is administered once every three weeks. [16A] The cancer is selected from the group consisting of renal cell carcinoma, clear cell renal carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, thyroid cancer, cholangiocarcinoma, lung cancer, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, or neuroblastoma, the antibody-drug conjugate according to any one of [1A] to [15A]. [17A] The antibody-drug conjugate according to [16A], wherein the cancer is renal cell carcinoma. [18A] The antibody-drug conjugate according to [16A], wherein the cancer is ovarian cancer. [19A] The antibody-drug conjugate according to any one of [1A] to [18A], wherein the cancer is resistant or refractory. [20A] The resistance or refractoriness is resistance or refractoriness obtained by the cancer due to treatment with an anticancer drug, the antibody-drug conjugate according to [19A]. [21A] The anti-cancer drug is the antibody-drug conjugate described in [20A], which is a platinum-based chemotherapeutic agent, chemotherapy, PARP inhibitor, immune checkpoint inhibitor, angiogenesis inhibitor, or VEGFR-TKI. [22A] The anti-cancer drug is a platinum-based chemotherapeutic agent, which is the antibody-drug conjugate described in [20A]. [23A] The platinum-based chemotherapeutic agent is the antibody-drug conjugate described in [22A], which includes a platinum-based drug and a taxane. [24A] The cancer is a CDH6-expressing cancer, which is the antibody-drug conjugate described in any one of [1A] to [23A]. [25A] The CDH6-expressing cancer is a CDH6-overexpressing cancer, which is the antibody-drug conjugate described in [24A]. [26A] The cancer is an inoperable or recurrent cancer, which is the antibody-drug conjugate described in any one of [1A] to [25A]. [27A] A pharmaceutical composition containing the antibody-drug conjugate described in any one of [1A] to [26A] or a salt thereof as an active ingredient and a pharmaceutically acceptable formulation ingredient. [28A] A method for treating or preventing cancer in a subject, which includes administering to the subject an anti-CDH6 antibody-drug conjugate containing an anti-CDH6 antibody and an anti-tumor compound connected by a linker, wherein the linker and the anti-tumor compound have the following formula: -(Succinimid-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GGFG-NH-CH2-O-CH2-C(=O)-(NH-DX) is represented by wherein, -(Succinimid-3-yl-N)- has the structure represented by the following formula:

[0031]

Chemical formula

[0032] [Chemical formula] represents a group represented by, the nitrogen atom of the amino group at the 1-position is the connecting position, and the anti-CDH6 antibody, in its heavy chain variable region, has CDRH1 consisting of the amino acid sequence of SEQ ID NO: 17, CDRH2 consisting of the amino acid sequence of SEQ ID NO: 60, and CDRH3 consisting of the amino acid sequence of SEQ ID NO: 19, and in its light chain variable region, has CDRL1 consisting of the amino acid sequence of SEQ ID NO: 12, CDRL2 consisting of the amino acid sequence of SEQ ID NO: 13, and CDRL3 consisting of the amino acid sequence of SEQ ID NO: 14, method. [29A] The antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 71 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 63, the method according to [28A]. [30A] The antibody-drug conjugate has the following formula:

[0033] [Chemical formula] has a structure represented by, wherein Ab is an antibody comprising a heavy chain variable region comprising SEQ ID NO: 71 and a light chain variable region comprising SEQ ID NO: 63, n represents the average number of units of the drug-linker structure conjugated to the antibody per antibody, and the average number of units of the conjugated selected drug-linker structure per antibody is in the range of 1 to 10, the method according to [28A] or [29A]. [31A] The antibody comprises a heavy chain comprising the amino acid sequence at positions 20 to 471 in SEQ ID NO: 69 and a light chain comprising the amino acid sequence at positions 21 to 233 in SEQ ID NO: 61, the method according to any one of [28A] to [30A]. [32A]The heavy or light chain has undergone one or more types of modifications selected from the group consisting of N-linked glycosylation, O-linked glycosylation, N-terminal processing, C-terminal processing, deamidation, isomerization of aspartic acid, oxidation of methionine, addition of a methionine residue to the N-terminus, amidation of a proline residue, conversion of the N-terminal glutamine or N-terminal glutamic acid to pyroglutamic acid, and deletion of one or two amino acids from the carboxyl terminus, the method according to any one of [28A] to [31A]. [33A]The anti-CDH6 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain, the method according to any one of [28A] to [32A]. [34A]The average number of units of conjugated anti-tumor compound per antibody is in the range of 2 to 8, the method according to any one of [28A] to [33A]. [35A]The average number of units of conjugated anti-tumor compound per antibody is 7 to 8, the method according to any one of [28A] to [33A]. [36A]A dose of antibody-drug conjugate in the range of 1.6 mg / kg to 9.6 mg / kg is administered to a subject having cancer, the method according to any one of [28A] to [35A]. [37A]A dose of about 3.2 mg / kg of antibody-drug conjugate is administered to a subject having cancer, the method according to any one of [28A] to [35A]. [38A]A dose of about 4.8 mg / kg of antibody-drug conjugate is administered to a subject having cancer, the method according to any one of [28A] to [35A]. [39A]A dose of about 6.4 mg / kg of antibody-drug conjugate is administered to a subject having cancer, the method according to any one of [28A] to [35A]. [40A]A dose of about 8.0 mg / kg of antibody-drug conjugate is administered to a subject having cancer, the method according to any one of [28A] to [35A]. [41A]The antibody-drug conjugate is administered by intravenous administration, the method according to any one of [28A] to [40A]. [42A] The method according to any one of [28A] to [41A], wherein the antibody-drug conjugate is administered once every three weeks. [43A] The method according to any one of [28A] to [42A], wherein the cancer is selected from the group consisting of renal cell carcinoma, clear cell renal carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, thyroid cancer, cholangiocarcinoma, lung cancer, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, or neuroblastoma. [44A] The method according to [43A], wherein the cancer is renal cell carcinoma. [45A] The method according to [43A], wherein the cancer is ovarian cancer. [46A] The method according to any one of [28A] to [45A], wherein the cancer is resistant or refractory. [47A] The method according to [46A], wherein the resistance or refractoriness is resistance or refractoriness obtained by the cancer due to treatment with an anticancer drug. [48A] The method according to [47A], wherein the anticancer drug is a platinum-based chemotherapeutic agent, chemotherapy, PARP inhibitor, immune checkpoint inhibitor, angiogenesis inhibitor, or VEGFR-TKI. [49A] The method according to [47A], wherein the anticancer drug is a platinum-based chemotherapeutic agent. [50A] The method according to [49A], wherein the platinum-based chemotherapeutic agent includes a platinum-based drug and a taxane. [51A] The method according to any one of [28A] to [50A], wherein the cancer is CDH6-expressing cancer. [52A] The method according to [51A], wherein the CDH6-expressing cancer is CDH6-overexpressing cancer. [53A] The method according to any one of [28A] to [52A], wherein the cancer is inoperable or recurrent cancer. [54A] The method according to any one of [28A] to [53A], wherein the antibody-drug conjugate is administered in the form of a pharmaceutical composition containing at least one pharmaceutically acceptable formulation component. Use of an anti-CDH6 antibody-drug conjugate in the manufacture of a medicament for treating or preventing cancer, wherein the antibody-drug conjugate comprises an anti-CDH6 antibody and an anti-tumor compound connected by a linker, and the linker and the anti-tumor compound have the following formula: -(Succinimid-3-yl-N)-CH2CH2CH2CH2CH2-C(=O)-GGFG-NH-CH2-O-CH2-C(=O)-(NH-DX) represented by wherein -(Succinimid-3-yl-N)- has the structure represented by the following formula:

[0034]

Chemical formula

[0035]

Chemical formula

[0036]

Chemical formula

[0037] In some embodiments, the present disclosure provides an anti-CDH6 antibody-drug conjugate for use in treating or preventing cancer, wherein the antibody-drug conjugate comprises an anti-CDH6 antibody and an anti-tumor compound connected by a linker.

[0038] In some embodiments, the present disclosure provides a method of treating or preventing cancer in a subject, comprising administering to the subject an anti-CDH6 antibody-drug conjugate comprising an anti-CDH6 antibody and an anti-tumor compound connected by a linker.

[0039] In some embodiments, the present disclosure provides the use of an anti-CDH6 antibody-drug conjugate in the manufacture of a medicament for treating or preventing cancer, wherein the antibody-drug conjugate comprises an anti-CDH6 antibody and an anti-tumor compound connected by a linker.

[0040] In some embodiments, the present disclosure provides a pharmaceutical composition for treating or preventing cancer, comprising an anti-CDH6 antibody-drug conjugate as an active ingredient, wherein the antibody-drug conjugate comprises an anti-CDH6 antibody and an anti-tumor compound connected by a linker.

[0041] In some embodiments, the linker and the anti-tumor compound have the following formula: -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-(NH-DX) represented by wherein, -(Succinimid-3-yl-N)- is the following formula:

[0042]

Chemical formula

[0043]

Chemical formula

[0044] In some embodiments, the anti-CDH6 antibody comprises, in its heavy chain variable region, CDRH1 consisting of the amino acid sequence of SEQ ID NO: 17, CDRH2 consisting of the amino acid sequence of SEQ ID NO: 60, and CDRH3 consisting of the amino acid sequence of SEQ ID NO: 19, and in its light chain variable region, CDRL1 consisting of the amino acid sequence of SEQ ID NO: 12, CDRL2 consisting of the amino acid sequence of SEQ ID NO: 13, and CDRL3 consisting of the amino acid sequence of SEQ ID NO: 14.

[0045] In some embodiments, the anti-CDH6 antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 71 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 63.

[0046] In some embodiments, the antibody-drug conjugate is the following formula:

[0047]

Chemical formula

[0048] In some embodiments, the anti-CDH6 antibody comprises a heavy chain comprising the amino acid sequence at positions 20 to 471 in SEQ ID NO: 69 and a light chain comprising the amino acid sequence at positions 21 to 233 in SEQ ID NO: 61.

[0049] In some embodiments, the heavy chain or the light chain has undergone two or more modifications selected from the group consisting of N-linked glycosylation, O-linked glycosylation, N-terminal processing, C-terminal processing, deamidation, asparagine isomerization, methionine oxidation, addition of a methionine residue to the N-terminus, amidation of a proline residue, conversion of the N-terminal glutamine or N-terminal glutamate to pyroglutamic acid, and deletion of one or two amino acids from the carboxyl terminus.

[0050] In some embodiments, the anti-CDH6 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain.

[0051] In some embodiments, the average number of units of the conjugated selected drug-linker structure per antibody is in the range of 1 to 10. In some embodiments, the average number of units of the conjugated selected drug-linker structure per antibody is in the range of 2 to 8. In some embodiments, the average number of units of the conjugated selected drug-linker structure per antibody is in the range of 5 to 8. In some embodiments, the average number of units of the conjugated selected drug-linker structure per antibody is in the range of 7 to 8.

[0052] In some embodiments, a dose of the antibody-drug conjugate in the range of 1.6 mg / kg to 9.6 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate in the range of 3.2 mg / kg to 9.6 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate in the range of 4.8 mg / kg to 9.6 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate in the range of 3.2 mg / kg to 8.0 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate in the range of 3.2 mg / kg to 6.4 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate in the range of 4.8 mg / kg to 8.0 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate in the range of 4.8 mg / kg to 6.4 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate of about 3.2 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate of about 4.8 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate of about 5.4 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate of about 5.6 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate of about 6.4 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate of about 8.0 mg / kg is administered to a subject having cancer. In some embodiments, a dose of the antibody-drug conjugate of about 9.6 mg / kg is administered to a subject having cancer.

[0053] In some embodiments, a dose of the antibody-drug conjugate of 3.2 mg / kg, 3.3 mg / kg, 3.4 mg / kg, 3.5 mg / kg, 3.6 mg / kg, 3.7 mg / kg, 3.8 mg / kg, 3.9 mg / kg, 4.0 mg / kg, 4.1 mg / kg, 4.2 mg / kg, 4.3 mg / kg, 4.4 mg / kg, 4.5 mg / kg, 4.6 mg / kg, 4.7 mg / kg, 4.8 mg / kg, 4.9 mg / kg, 5.0 mg / kg, 5.1 mg / kg, 5.2 mg / kg, 5.3 mg / kg, 5.4 mg / kg, 5.5 mg / kg, 5.6 mg / kg, 5.7 mg / kg, 5.8 mg / kg, 5.9 mg / kg, 6.0 mg / kg, 6.1 mg / kg, 6.2 mg / kg, 6.3 mg / kg, or 6.4 mg / kg is administered to a subject having cancer.

[0054] In some embodiments, the antibody-drug conjugate is administered by intravenous administration.

[0055] In some embodiments, the antibody-drug conjugate is administered once every three weeks.

[0056] In some embodiments, the cancer is selected from the group consisting of renal cell carcinoma, clear cell renal cell carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, clear cell carcinoma of the ovary, endometrioid carcinoma of the ovary, ovarian mucinous tumor, thyroid cancer, cholangiocarcinoma, lung cancer, non-small cell lung cancer, cervical cancer, brain tumor, head and neck cancer, sarcoma, osteosarcoma, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, neuroblastoma, colorectal cancer, gastric cancer, endometrial cancer, nasopharyngeal cancer, prostate cancer, or cancer associated with von Hippel-Lindau disease. In some embodiments, the cancer is selected from the group consisting of renal cell carcinoma, clear cell renal cell carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, thyroid cancer, cholangiocarcinoma, lung cancer, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, and neuroblastoma. In some embodiments, the cancer is ovarian cancer. In some embodiments, the ovarian cancer is epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal cancer. In some embodiments, the cancer is metastatic. In some embodiments, the cancer is renal cell carcinoma, clear cell renal cell carcinoma, or papillary renal cell carcinoma. In some embodiments, the cancer is renal cell carcinoma.

[0057] In some embodiments, the cancer is resistant or refractory. In some embodiments, the resistance or refractoriness is resistance or refractoriness acquired by the cancer due to treatment with an anti-cancer drug. In some embodiments, the anti-cancer drug is a platinum-based chemotherapeutic agent, chemotherapeutic agent, poly ADP-ribose polymerase (PARP) inhibitor, immune checkpoint inhibitor (ICI), angiogenesis inhibitor, or vascular endothelial growth factor-tyrosine kinase inhibitor (VEGFR-TKI). In some embodiments, the anti-cancer drug is a platinum-based chemotherapeutic agent. In some embodiments, the platinum-based chemotherapeutic agent includes a platinum-based drug and a taxane.

[0058] In some embodiments, the subject has cancer that is resistant to platinum-based chemotherapy. In some embodiments, the subject exhibits cancer recurrence prior to administration of the ADC. In some embodiments, the subject has cancer that is resistant or refractory to a chemotherapy regimen that includes a platinum-based drug and a taxane.

[0059] In some embodiments, cancer recurrence occurs less than or within about 6 months after completion of a chemotherapy regimen that includes a platinum-based drug. In some embodiments, cancer recurrence occurs less than about 6 months after completion of a chemotherapy regimen that includes a platinum-based drug. In some embodiments, cancer recurrence occurs within about 6 months after completion of a chemotherapy regimen that includes a platinum-based drug.

[0060] In some embodiments, cancer recurrence occurs less than or within about 6 months after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane. In some embodiments, cancer recurrence occurs less than about 6 months after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane. In some embodiments, cancer recurrence occurs within about 6 months after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane.

[0061] In some embodiments, cancer recurrence occurs about 6 months after or after about 6 months have elapsed after completion of a chemotherapy regimen that includes a platinum-based drug. In some embodiments, cancer recurrence occurs about 6 months after completion of a chemotherapy regimen that includes a platinum-based drug. In some embodiments, cancer recurrence occurs after about 6 months have elapsed after completion of a chemotherapy regimen that includes a platinum-based drug.

[0062] In some embodiments, cancer recurrence occurs about 6 months after or after about 6 months have elapsed after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane. In some embodiments, cancer recurrence occurs about 6 months after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane. In some embodiments, cancer recurrence occurs after about 6 months have elapsed after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane.

[0063] In some embodiments, cancer recurrence occurs less than or within 6 months after completion of a chemotherapy regimen that includes a platinum-based drug. In some embodiments, cancer recurrence occurs less than 6 months after completion of a chemotherapy regimen that includes a platinum-based drug. In some embodiments, cancer recurrence occurs within 6 months after completion of a chemotherapy regimen that includes a platinum-based drug.

[0064] In some embodiments, cancer recurrence occurs less than or within 6 months after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane. In some embodiments, cancer recurrence occurs less than 6 months after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane. In some embodiments, cancer recurrence occurs within 6 months after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane.

[0065] In some embodiments, cancer recurrence occurs 6 months or more after completion of a chemotherapy regimen that includes a platinum-based drug. In some embodiments, cancer recurrence occurs 6 months after completion of a chemotherapy regimen that includes a platinum-based drug. In some embodiments, cancer recurrence occurs 6 months or more after completion of a chemotherapy regimen that includes a platinum-based drug.

[0066] In some embodiments, cancer recurrence occurs 6 months or more after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane. In some embodiments, cancer recurrence occurs 6 months after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane. In some embodiments, cancer recurrence occurs 6 months or more after completion of a chemotherapy regimen that includes a platinum-based drug and a taxane.

[0067] In some embodiments, the subject has cancer that is resistant or refractory to poly(ADP-ribose) polymerase (PARP) inhibitors. In some embodiments, the subject has cancer that is resistant or refractory to immune checkpoint inhibitors (ICIs). In some embodiments, the subject has cancer that is resistant or refractory to angiogenesis inhibitors. In some embodiments, the subject has cancer that is resistant or refractory to vascular endothelial growth factor-tyrosine kinase inhibitors (VEGFR-TKIs).

[0068] In some embodiments, the subject has a treatment history with one or more types of anti-cancer drugs selected from the group consisting of platinum-based chemotherapeutic agents, chemotherapeutic agents, PARP inhibitors, immune checkpoint inhibitors, angiogenesis inhibitors, and VEGFR-TKIs.

[0069] In some embodiments, the cancer comprises one or more tumors that express CDH6. In some embodiments, the CDH6-expressing cancer is a CDH6-overexpressing cancer. In some embodiments, the CDH6-overexpressing cancer is a cancer that is given a high score for the expression of CDH6 in immunohistochemistry. In some embodiments, the CDH6-overexpressing cancer is a cancer that is given a high score for the expression of CDH6 in in situ hybridization. In some embodiments, the CDH6-overexpressing cancer is a cancer that is given a high score for the expression of CDH6 in next-generation sequencing.

[0070] In some embodiments, the cancer is inoperable or recurrent cancer.

[0071] In some embodiments, the objective response rate of a subject administered a therapeutically effective amount of the ADC is at least about 20%. In some embodiments, the objective response rate of a subject administered a therapeutically effective amount of the ADC is at least about 30%. In some embodiments, the objective response rate of a subject administered a therapeutically effective amount of the ADC is at least about 40%. In some embodiments, the objective response rate of a subject administered a therapeutically effective amount of the ADC is at least about 50%. In some embodiments, the objective response rate of a subject administered a therapeutically effective amount of the ADC is at least about 60%.

[0072] In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 5 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 5.5 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 5.6 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 5.8 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 6.0 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 7.0 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 7.9 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 8 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 9 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 10 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 11 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 12 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 13 months. In some embodiments, the progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 13.9 months.

[0073] In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 5 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 5.5 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 5.6 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 5.8 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 6.0 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 7.0 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 7.9 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 8 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 9 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 10 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 11 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 12 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 13 months. In some embodiments, the median progression-free survival period of a subject administered a therapeutically effective amount of the ADC is at least about 13.9 months.

[0074] In some embodiments, the ADC is administered to the subject as a monotherapy.

[0075] In some embodiments, the subject is administered the ADC together with a second drug. In some embodiments, the ADC is administered before the second drug. In some embodiments, the ADC is administered after the second drug. In some embodiments, the ADC is administered simultaneously with the second drug.

[0076] In some embodiments, the ADC is administered to a subject as maintenance therapy.

[0077] In some embodiments, the ADC is administered to a subject as adjuvant therapy. In some embodiments, the ADC is administered to a subject after surgical resection of a tumor.

[0078] In some embodiments, the ADC is administered to a subject as neoadjuvant therapy. In some embodiments, the ADC is administered to a subject before surgical resection of a tumor.

[0079] In some embodiments, the present disclosure generally relates to an anti-CDH6 antibody-drug conjugate (ADC) for use in treating or preventing cancer, wherein the ADC has the following formula:

[0080]

Chemical formula

[0081] Also provided herein is a pharmaceutical composition comprising an antibody-drug conjugate or a salt thereof according to any one of the foregoing embodiments or embodiments as an active ingredient and a pharmaceutically acceptable formulation ingredient.

[0082] The foregoing schematic description and the following detailed description are exemplary and explanatory and are intended to provide a further description of the claimed present disclosure. Other objectives, advantages, and novel features will be readily apparent to those skilled in the art from the following brief description of the drawings and the detailed description of the present disclosure.

Brief Description of the Drawings

[0083]

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Mode for Carrying Out the Invention

[0084] Description of Embodiments Hereinafter, preferred embodiments for carrying out the disclosed methods and uses will be described with reference to the drawings. It should be noted that the embodiments described below are merely illustrative of representative embodiments, and the scope of the present disclosure should not be construed narrowly for these examples.

[0085] It should be understood that the method is not limited to the specific embodiments described and may therefore vary. It should also be understood that the terms used herein are for the purpose of describing only specific embodiments and are not intended to be limiting. The scope of the present technology will be limited only by the appended claims.

[0086] 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. Although any methods and materials similar or equivalent to those described herein may be used in the practice or testing of the disclosed methods and uses, representative exemplary methods and materials are described herein.

[0087] When ranges of values are provided, each intervening value between the upper and lower limits of the range, to the tenth of the unit of the lower limit, and any other stated value or intervening value within the stated range, is included within the disclosure, unless otherwise clearly indicated in the context. The upper and lower limits of these smaller ranges may independently be included within the smaller ranges and are also included within the disclosure, subject to any specifically excluded limits in the stated range. When the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included within the disclosure.

[0088] As used in this specification and the claims, the singular forms “a,” “an,” and “the” include the singular and plural referents unless the context clearly dictates otherwise.

[0089] As used in this specification, the term “comprising” is intended to mean that the compositions and methods include the recited elements but do not exclude others. When used to define compositions and methods, “consisting essentially of” is intended to mean excluding other elements that are essential to the composition or method. “Consisting of” is intended to mean excluding more than trace amounts of other components with respect to the claimed compositions and substantial method steps. Embodiments defined by each of these transitional terms are within the scope of the disclosure. Thus, methods and compositions can include (comprising) additional steps and components, or alternatively, include (consisting essentially of) non-essential steps and compositions, or alternatively, are intended to include (consisting of) only the recited method steps or compositions.

[0090] As used in this specification, “about” also means plus or minus 10% of the specified number. For example, “about 10” should be understood as both “10” and “9 - 11”.

[0091] As used herein, "optional" or "optionally" means that the event or circumstance described thereafter may or may not occur, and that the description includes both the case where the event or circumstance occurs and the case where it does not occur.

[0092] The terms "individual", "subject", and "patient" are used interchangeably herein and refer to any individual mammal, such as a cow, dog, cat, horse, primate, pig, camel, bat, or human, that is treated according to the disclosed methods or uses. In a preferred embodiment, the subject is a human.

[0093] As used herein, the phrases "effective amount", "therapeutically effective amount", and "therapeutic level" mean, for that purpose, a dosage or concentration in a subject that provides a specific pharmacological effect for which such treatment is required, i.e., to treat or prevent cancer (e.g., renal cell carcinoma, ovarian cancer, CDH6-expressing cancer, or resistant or refractory cancer) for which an ADC is administered to the subject. It is emphasized that a therapeutically effective amount or therapeutic level of an ADC may not necessarily be effective in treating the cancers described herein, even if such a dosage is considered by one of ordinary skill in the art to be a therapeutically effective amount. Merely for convenience, exemplary dosages, drug delivery amounts, therapeutically effective amounts, and therapeutic levels are provided below. One of ordinary skill in the art can adjust such amounts as needed, according to standard practice, to treat a specific subject and / or medical condition. The therapeutically effective amount can vary based on the route and dosage form of administration, the age and weight of the subject, and / or the medical condition of the subject, including the type and severity of the cancer.

[0094] As used herein in the context of cancer, the terms "treatment" or "treating" refer to reducing, suppressing, or eliminating cancer; reducing, suppressing, or eliminating cancer cell growth; reducing, suppressing, or eliminating cancer spread; or shrinking or killing a tumor or metastasis. Treatment and treating can optionally also mean improving the quality of life or overall survival of a subject, even if cancer cell growth is not inhibited and / or the cancer does not die.

[0095] As used herein in the context of cancer, the terms "prevent" or "preventing" refer to delaying or preventing the occurrence of metastasis (i.e., the growth of cancer at a secondary site where cancer was not present at the start of treatment), and, when a subject has reached remission or the cancer / tumor has been completely destroyed or killed, delaying or preventing cancer recurrence.

[0096] As used herein, the term "pharmaceutical composition" refers to a combination of an active agent and an inert or active carrier that renders the composition particularly suitable for diagnostic or therapeutic use in vivo or ex vivo.

[0097] As used herein, the term "pharmaceutically acceptable carrier" refers to any of the standard pharmaceutical carriers such as phosphate buffered saline solution, water, emulsions (e.g., oil / water or water / oil emulsions, etc.), and various types of wetting agents. The composition can also include stabilizers and preservatives. For examples of carriers, stabilizers, and adjuvants, see, e.g., Martin, Remington’s Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA

[1975] .

[0098] As used herein, the terms "parenteral administration" and "administered parenterally" mean modes of administration other than enteral and topical administration, typically by injection, including, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0099] As used herein, the terms "systemic administration", "administered systemically", "peripheral administration", and "administered peripherally" mean administration of a compound, drug, or other material other than directly to the central nervous system, such that the compound, drug, or other material enters the patient's body and is thus subject to metabolism and other similar processes, for example subcutaneous administration.

[0100] In this description, the term "cancer" is used to have the same meaning as the term "tumor".

[0101] In this description, the term "gene" is used to include not only DNA, but also its mRNA and cDNA, and its cRNA.

[0102] In this description, the terms "polynucleotide" or "nucleotide" are used to have the same meaning as nucleic acid, and include DNA, RNA, probes, oligonucleotides, and primers. In this description, the terms "polynucleotide" and "nucleotide" may be used interchangeably with each other unless otherwise specified.

[0103] In this description, the terms "polypeptide" and "protein" may be used interchangeably with each other.

[0104] In this description, the term "cell" includes cells in individual animals and cultured cells.

[0105] In this description, the term "CDH6" may be used to have the same meaning as that of the CDH6 protein. In this description, human CDH6 is also referred to as "hCDH6".

[0106] In this description, the term "cytotoxic activity" is used to mean that pathological changes are caused to cells by any given means. The term means not only direct trauma, but also all types of structural or functional damage caused to cells, such as DNA cleavage, formation of base dimers, chromosomal breakage, damage to the cell mitotic apparatus, and decrease in the activity of various types of enzymes.

[0107] In this description, the phrase "exhibiting toxicity in cells" is used to mean that toxicity is presented in cells by any given means. The term means not only direct trauma, but also all types of structural, functional, or metabolic effects caused to cells, such as DNA cleavage, formation of base dimers, chromosomal breakage, damage to the cell mitotic apparatus, decrease in the activity of various types of enzymes, and suppression of the effects of cell growth factors.

[0108] In this description, the term "functional fragment of an antibody", also referred to as "antigen-binding fragment of an antibody", is used to mean a specific fragment of an antibody that has binding activity to an antigen, and includes Fab, F(ab’)2, scFv, diabody, linear antibody, and multispecific antibody formed from antibody fragments. Fab’, which is a monovalent fragment of the antibody variable region obtained by treating F(ab’)2 under reducing conditions, is also included in the antigen-binding fragment of the antibody. However, as long as the antigen-binding fragment has antigen-binding ability, the antigen-binding fragment of the antibody is not limited to these molecules. These antigen-binding fragments include not only those obtained by treating the full-length molecule of the antibody protein with an appropriate enzyme, but also proteins produced in appropriate host cells using genetically engineered antibody genes.

[0109] In this description, the term "epitope" is used to mean a partial peptide or partial three-dimensional structure of CDH6 to which a specific anti-CDH6 antibody binds. Such an epitope, which is the above-described partial peptide of CDH6, can be determined by methods well known to those skilled in the art, such as immunoassays. First, various partial structures of the antigen are produced. For the production of such partial structures, known oligopeptide synthesis techniques can be applied. For example, a series of polypeptides in which CDH6 is successively truncated by an appropriate length from its C-terminus or N-terminus are produced by genetic recombination techniques well known to those skilled in the art. Then, the reactivity of antibodies against such polypeptides is investigated to roughly determine the recognition site. Thereafter, shorter peptides are further synthesized, and then their reactivity against these peptides is investigated to determine the epitope. When an antibody that binds to a membrane protein having multiple extracellular domains is directed to a three-dimensional structure composed of multiple domains as an epitope, the domain to which the antibody binds can be determined by modifying the amino acid sequence of the specific extracellular domain and thereby modifying the three-dimensional structure. An epitope, which is a partial three-dimensional structure of an antigen that binds to a specific antibody, can also be determined by X-ray structural analysis by specifying the amino acid residues of the antigen adjacent to the antibody.

[0110] In this description, the phrase "antibodies that bind to the same epitope" is used to mean antibodies that bind to a common epitope. When a second antibody binds to a partial peptide or partial three-dimensional structure to which the first antibody binds, the first antibody and the second antibody can be determined to bind to the same epitope. Alternatively, by confirming that the second antibody competes with the first antibody for binding to the antigen (i.e., the second antibody inhibits the binding of the first antibody to the antigen), even if the specific sequence or structure of the epitope has not been determined, the first antibody and the second antibody can be determined to bind to the same epitope. In this description, the phrase "bind to the same epitope" refers to the case where, by either or both of these determination methods, it is determined that the first antibody and the second antibody bind to a common epitope. If the first antibody and the second antibody bind to the same epitope and, further, the first antibody has a special effect such as antitumor activity or internalization activity, the second antibody can be expected to have the same activity as that of the first antibody.

[0111] In this description, the term "CDR" is used to mean complementarity-determining region. It is known that the heavy chain and the light chain of an antibody molecule each have three CDRs. Such CDRs are also referred to as hypervariable regions and are located in the variable regions of the heavy chain and the light chain of the antibody. These regions have a particularly highly variable primary structure and are separated into three sites on the primary structure of the polypeptide chain in each of the heavy chain and the light chain. In this description, with respect to the CDRs of an antibody, the CDRs of the heavy chain are referred to as CDRH1, CDRH2, and CDRH3, respectively, from the amino-terminal side of the amino acid sequence of the heavy chain, while the CDRs of the light chain are referred to as CDRL1, CDRL2, and CDRL3, respectively, from the amino-terminal side of the amino acid sequence of the light chain. These sites are located close to each other in a three-dimensional structure and determine the specificity of the antibody for the antigen to which the antibody binds.

[0112] In the present disclosure, the phrase "hybridize under stringent conditions" is used to mean that hybridization is carried out at 68° C. in a commercially available hybridization solution, ExpressHyb hybridization solution (manufactured by Clontech Laboratories, Inc.), or that hybridization is carried out at 68° C. using a DNA immobilized filter in the presence of 0.7 to 1.0 M NaCl, and the resulting product is then washed with a 0.1 to 2-fold concentrated SSC solution (1-fold concentrated SSC consists of 150 mM NaCl and 15 mM sodium citrate) for identification, or hybridization is carried out under conditions equivalent thereto.

[0113] In this description, the term "one to several" is used to mean one to ten, one to nine, one to eight, one to seven, one to six, one to five, one to four, one to three, or one or two.

[0114] In this description, the term "anticancer drug" is used to mean a drug for the purpose of suppressing, preventing, inhibiting, and / or decelerating the proliferation, growth, and / or spread of cancer cells or tumor cells. In this description, the term "anticancer drug" has the same meaning as the terms "anticancer agent", "antineoplastic agent", or "antineoplastic drug".

[0115] In this description, the term "resistance" is used to mean having no efficacy against treatment with an anticancer drug. The term may also be expressed as "refractory", "inefficacy", or "non-responsiveness". Further, due to the reason that tumor growth cannot be blocked due to the inefficacy characteristics, the term may also be expressed as "intolerance". In this description, the term "resistance" is also used when the cancer of the subject exhibits low sensitivity to treatment with an anticancer drug, and after treatment with an anticancer drug, the cancer cells do not disappear or shrink, CR or PR is not achieved, and / or the cancer cells progress early (for example, within or after less than six months for ovarian cancer).

[0116] In this description, the term "resistance" can mean "having resistance acquired by cancer due to treatment with an anticancer drug" or "having resistance inherent in cancer independently of treatment with an anticancer drug".

[0117] In this description, the term "chemotherapy" is used to mean a therapy using one or more chemotherapeutic drugs used for treating cancer.

[0118] In this description, the term "chemotherapeutic drug" is used to mean a chemotherapeutic agent used for treating cancer. Chemotherapeutic drugs include, but are not limited to, alkylating agents (e.g., mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, hexamethylmelamine, thiotepa, busulfan, carmustine, lomustine, semustine, streptozocin, dacarbazine), antimetabolites (e.g., gemcitabine, methotrexate, fluorouracil, doxifluridine, capecitabine, floxuridine, cytarabine, mercaptopurine, thioguanine, pentostatin), vinca alkaloids (e.g., vinblastine, vincristine), epipodophyllotoxins (e.g., etoposide, teniposide), antibiotics (e.g., dactinomycin, daunorubicin, doxorubicin, bleomycin, plicamycin, mitomycin), platinum complexes (e.g., cisplatin, carboplatin, oxaliplatin), taxanes (e.g., paclitaxel, docetaxel), anthracenediones (e.g., mitoxantrone), substituted ureas (e.g., hydroxyurea), methylhydrazine (e.g., procarbazine hydrochloride), vitamin A metabolites (e.g., tretinoin), and pharmaceutically acceptable salts thereof.

[0119] In this description, the term "platinum-based chemotherapy" is used to mean a cancer therapy using one or more platinum-based drugs, with or without one or more other chemotherapeutic drugs.

[0120] In this description, the term "platinum-based chemotherapeutic agent" is used to mean a platinum-based drug or a series of drugs used in platinum-based chemotherapy.

[0121] In this description, the term "platinum-based drug" is used to mean a platinum complex used to treat cancer. Platinum-based drugs include, but are not limited to, cisplatin, carboplatin, and oxaliplatin.

[0122] In this description, the term "PARP inhibitor" is used to mean a drug that has the function of inhibiting PARP (poly[adenosine-5'-diphosphate (ADP)-ribose] polymerase) and thus prevents single-strand break repair (Benafif S et al., Onco.Targets Ther. (2015) 8, 519-528.) (Fong PC et al., N.Engl.J.Med. (2009) 361, 123-134.) (Gelmon KA et al., Lancet Oncol. (2011) 12, 852-861.). PARP includes multiple subtypes, but the PARP inhibitor in the present invention preferably inhibits PARP-1 and PARP-2. PARP inhibitors include, but are not limited to, olaparib (Menear KA et al., J.Med.Chem. (2008) 51, 6581-6591.), rucaparib (Gillmore AT et al., Org.Process Res.Dev. (2012) 16, 1897-1904.), niraparib (Jones P et al., J.Med.Chem. (2009) 52, 7170-7185.), talazoparib (Shen Y et al., Clin.Cancer Res. (2013) 19(18), 5003-5015.), veliparib, pamiparib, and fluzoparib, as well as pharmaceutically acceptable salts thereof. Olaparib, rucaparib, niraparib, talazoparib, and veliparib, as well as pharmaceutically acceptable salts thereof, may preferably be exemplified.

[0123] In this description, the term "immune checkpoint inhibitor" is used to mean an agent that inhibits the immunosuppressive system and activates tumor immunity. Immune checkpoint inhibitors include, but are not limited to, anti-PD-1 antibodies, anti-PD-L1 antibodies, and anti-CTLA-4 antibodies. Anti-PD-1 antibodies and anti-PD-L1 antibodies can preferably be exemplified.

[0124] In this description, the term "anti-PD-1 antibody" is used to mean an antibody or a functional fragment of an antibody that specifically binds to PD-1 (programmed cell death-1; CD279; PDCD1), and the antibody has the activity of reducing, inhibiting, and / or preventing signal transduction caused by the interaction between PD-1 and PD-L1 or PD-L2 as a binding partner. Anti-PD-1 antibodies include, but are not limited to, nivolumab (International Publication No. WO2006 / 121168, etc.), pembrolizumab (International Publication No. WO2008 / 156712, etc.), tislelizumab, sintilimab, spartalizumab, and semiprimab.

[0125] In this description, the term "anti-PD-L1 antibody" is used to mean an antibody or a functional fragment of an antibody that specifically binds to PD-L1 (programmed cell death ligand 1; CD274; B7-H1), and the antibody has the activity of reducing, inhibiting, and / or preventing signal transduction caused by the interaction between PD-L1 and PD-1 or B7.1 (CD80) as a binding partner. Anti-PD-L1 antibodies include, but are not limited to, atezolizumab (International Publication No. WO2010 / 077634, etc.), durvalumab (International Publication No. WO2011 / 066389, etc.), and avelumab (International Publication No. WO2013 / 079174, etc.).

[0126] In this description, the term "anti-CTLA-4 antibody" is used to mean an antibody or a functional fragment of an antibody that specifically binds to CTLA-4 (cytotoxic T lymphocyte-associated protein 4; CD152), and the antibody has an activity of reducing, inhibiting, and / or preventing signal transduction caused by the interaction between CTLA-4 and B7.1 (CD80) or B7.2 (CD86) as a binding partner. Anti-CTLA-4 antibodies include, but are not limited to, ipilimumab (International Publication No. WO2001 / 014424, etc.) and tremelimumab (International Publication No. WO2000 / 037504, etc.).

[0127] In this description, the term "angiogenesis inhibitor" is used to mean an agent that directly or indirectly inhibits the formation and / or growth of new blood vessels, regardless of the mechanism. VEGF (vascular endothelial growth factor) inhibitors can preferably be exemplified.

[0128] In this description, the term "VEGF inhibitor" is used to mean an agent that inhibits the interaction between VEGF and a VEGF receptor. VEGF inhibitors include, but are not limited to, anti-VEGF antibodies, anti-VEGF receptor antibodies, fusion proteins containing the extracellular domain of a VEGF receptor, and multispecific antibodies having binding specificity for VEGF.

[0129] In this description, the term "anti-VEGF antibody" is used to mean an antibody or a functional fragment of an antibody that specifically binds to VEGF. Anti-VEGF antibodies include, but are not limited to, bevacizumab and sevacizumab. Bevacizumab can preferably be exemplified.

[0130] In this description, the term "anti-VEGF receptor antibody" is used to mean an antibody or a functional fragment of an antibody that specifically binds to a VEGF receptor. Anti-VEGF receptor antibodies include, but are not limited to, ramucirumab.

[0131] In this description, the term "fusion protein containing the extracellular domain of a VEGF receptor" is used to mean a fusion protein containing a binding domain for VEGF, wherein the binding domain is derived from the extracellular domain of a VEGF receptor. The fusion protein containing the extracellular domain of a VEGF receptor includes, but is not limited to, aflibercept.

[0132] In this description, the term "multispecific antibody having binding specificity for VEGF" is used to mean an antibody containing binding specificities for at least two different sites, wherein one of the binding specificities is for VEGF and one or more others are for any other antigen. The multispecific antibody having binding specificity for VEGF includes, but is not limited to, ibonesimab (Esfandiari et al., Nat. Rev. Drug Discov. (2022) 21(6):411-412), CTX-009 (also referred to as ABL-001, Xu et al., Cancer Lett. (2022) 538:215699), BI836880 (Clin. Exp. Metastasis (2020) 37(6):637-648), navicixizumab (Esfandiari et al., Nat. Rev. Drug Discov. (2022) 21(6):411-412), and vanucizumab (Xu et al., Cancer Lett. (2022) 538:215699).

[0133] In this description, the term "VEGFR-TKI" is used to mean an agent that inhibits the tyrosine kinase of vascular endothelial growth factor receptor (VEGFR). A VEGFR-TKI may have an effect of inhibiting kinases other than VEGFR tyrosine kinase. VEGFR-TKIs include, but are not limited to, regorafenib, sorafenib, vandetanib (U.S. Patent No. 7,173,038), sunitinib (U.S. Patent No. 6,573,293), axitinib (U.S. Patent No. 6,534,524), pazopanib (U.S. Patent No. 7,105,530), lenvatinib (U.S. Patent No. 7,253,286), nintedanib (hereinafter also referred to as "BIBF1120") (U.S. Patent No. 6,762,180), cabozantinib (U.S. Patent No. 7,579,473), tivozanib (International Publication No. WO2002 / 088110), brivanib (International Publication No. WO2004 / 009784), linifanib (International Publication No. WO2014 / 022975), lucitanib (International Publication No. WO2008 / 112408), irrotinib, and ENMD-2076, and pharmaceutically acceptable salts thereof.

[0134] In this description, the term "cancer recurrence" is used to mean the return of cancer to the same location as the primary tumor or to another location in the body after a period during which the cancer could not be detected. The term is defined based on "recurrence" in the following references. NCI Dictionaries, "recurrence", NCI Dictionary of Cancer Terms [online]. National Cancer Institute [searched on 2022-09-06]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / recurrence>.

[0135] In this description, the term "chemotherapy regimen" is used to mean a treatment plan for chemotherapy that defines drugs, dosages, frequencies, etc.

[0136] In this description, the term "complete response (CR)" is used to mean that all signs of cancer have disappeared in response to treatment. "Complete response (CR)" does not necessarily mean that the cancer has been cured. The term may also be expressed as "complete remission". The term is defined based on "complete response" in the following references. NCI Dictionaries, "complete response", NCI Dictionary of Cancer Terms [online]. National Cancer Institute [searched on 2022-09-06]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / complete-response>.

[0137] In this description, the term "partial response (PR)" is used to mean that the size of the tumor or the extent of cancer in the body has decreased in response to treatment. The term may also be expressed as "partial remission". The term is defined based on "partial response" in the following references. NCI Dictionaries, "partial response", NCI Dictionary of Cancer Terms [online]. National Cancer Institute [searched on 2022-09-06]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / partial-response>.

[0138] In this description, the term "stable disease (SD)" is used to mean that the extent or severity of cancer has neither decreased nor increased. The term is defined based on "stable disease" in the following references. NCI Dictionaries, "stable disease", NCI Dictionary of Cancer Terms [online]. National Cancer Institute [searched on 2022-09-06]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / stable-disease>.

[0139] In this description, the term "progressive disease" or "PD" is used to mean cancer that is growing, spreading, or worsening. The term is defined based on "progressive disease" in the following references. NCI Dictionaries, "progressive disease", NCI Dictionary of Cancer Terms [online]. National Cancer Institute [searched on 2023-04-02]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / progressive-disease>.

[0140] In this description, the term "objective response rate" or "ORR" is used to mean the percentage of people in an investigational or treatment group who have a partial response (PR) or complete response (CR) to treatment within a specific period. The term is defined based on "ORR" in the following references. NCI Dictionaries, "ORR", NCI Dictionary of Cancer Terms [online]. National Cancer Institute [searched on 2023-04-02]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / orr>.

[0141] In this description, the term "progression-free survival" or "PFS" is used to mean the length of time during and after the treatment of a disease such as cancer, during which the patient lives with the disease without it getting worse. The term is defined based on "PFS" in the following references. NCI Dictionaries, "PFS", NCI Dictionary of Cancer Terms [Online]. National Cancer Institute [searched on 2023-04-02]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / pfs>.

[0142] In this description, the term "maintenance therapy" is used to mean a treatment given after initial therapy to help prevent the cancer from coming back after it has disappeared. The term is defined based on "maintenance therapy" in the following references. NCI Dictionaries, "maintenance therapy", NCI Dictionary of Cancer Terms [Online]. National Cancer Institute [searched on 2023-04-10]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / maintenance-therapy>.

[0143] In this description, the term "adjuvant therapy" is used to mean additional cancer treatment given after primary treatment to lower the risk that the cancer will come back. The term is defined based on "adjuvant therapy" in the following references. NCI Dictionaries, "adjuvant therapy", NCI Dictionary of Cancer Terms [online]. National Cancer Institute [searched on 2023-04-10]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / adjuvant-therapy>.

[0144] In this description, the term "neoadjuvant therapy" is used to mean a treatment given as a first step to shrink a tumor before the main treatment such as surgery is given. The term is defined based on "neoadjuvant therapy" in the following references. NCI Dictionaries, "neoadjuvant therapy", NCI Dictionary of Cancer Terms [online]. National Cancer Institute [searched on 2023-04-10]. Retrieved from <cancer.gov / publications / dictionaries / cancer-terms / def / neoadjuvant-therapy>.

[0145] As amino acid substitutions in this specification, conservative amino acid substitutions are preferred. Conservative amino acid substitutions refer to substitutions that occur within a group of amino acids related to the amino acid side chain. Preferred groups of amino acids are as follows: acidic group (aspartic acid and glutamic acid); basic group (lysine, arginine, and histidine); nonpolar group (alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan); and uncharged polar family (glycine, asparagine, glutamine, cysteine, serine, threonine, and tyrosine). More preferred groups of amino acids are as follows: aliphatic hydroxyl group (serine and threonine); amide-containing group (asparagine and glutamine); aliphatic group (alanine, valine, leucine, and isoleucine); and aromatic group (phenylalanine, tryptophan, and tyrosine). Such amino acid substitutions are preferably carried out within a range that does not impair the properties of the substance having the original amino acid sequence.

[0146] Throughout this description, when a composition is described as having, including, or comprising specific components, or when a process and method are described as having, including, or comprising specific steps, additionally, it is contemplated that there are compositions of the present disclosure consisting essentially of or consisting of the recited components, and processes and methods according to the present disclosure consisting essentially of or consisting of the recited processing steps.

[0147] As a general matter, compositions specifying percentages are by weight unless otherwise specified. Further, when variables are not accompanied by definitions, previous definitions for the variables control.

[0148] 1.CDH6 Cadherins are glycoproteins present on the surface of cell membranes and function as cell-cell adhesion molecules or as signaling molecules involved in cell-cell interactions through calcium ion-dependent binding of their N-terminal extracellular domains. Classical cadherins are within the cadherin superfamily and are single-pass transmembrane proteins composed of five extracellular domains (EC domains), one transmembrane region, and an intracellular domain.

[0149] CDH6 (cadherin-6) is a single-pass transmembrane protein composed of 790 amino acids, which is classified into the type II cadherin family, and this protein has an N-terminal extracellular and a C-terminal intracellular domain. The human CDH6 gene was first cloned in 1995 (Non-Patent Document 1), and its sequence can be referred to, for example, under accession numbers NM_004932 and NP_004923 (NCBI).

[0150] The CDH6 protein used in the present disclosure can be directly purified from CDH6-expressing cells of humans or non-human mammals (e.g., rats, mice, or monkeys) and then used, or the cell membrane fraction of the aforementioned cells can be prepared and used as the CDH6 protein. Alternatively, CDH6 can also be obtained by synthesizing it in vitro or by causing host cells to produce CDH6 by genetic manipulation. According to such genetic manipulation, specifically, by incorporating CDH6 cDNA into a vector capable of expressing CDH6 cDNA and then synthesizing CDH6 in a solution containing enzymes, substrates, and energy materials necessary for transcription and translation, or by transforming other prokaryotic or eukaryotic host cells to cause them to express CDH6, the CDH6 protein can be obtained. Also, CDH6-expressing cells or cell lines expressing CDH6 based on the above genetic manipulation can be used to present the CDH6 protein. Alternatively, an expression vector incorporated with CDH6 cDNA can be directly administered to an animal to be immunized, and CDH6 can be expressed in the body of the animal thus immunized.

[0151] Furthermore, proteins consisting of amino acid sequences containing substitution, deletion, and / or addition of one or several amino acids in the above amino acid sequence of CDH6 and having biological activities equivalent to those of the CDH6 protein are also included within the term "CDH6".

[0152] The human CDH6 protein has the amino acid sequence shown in SEQ ID NO: 1. The extracellular region of the human CDH6 protein has extracellular domain 1 (also referred to as EC1 herein) having the amino acid sequence at positions 54 to 159 in the amino acid sequence shown in SEQ ID NO: 1, extracellular domain 2 (also referred to as EC2 herein) having the amino acid sequence at positions 160 to 268 in the amino acid sequence shown in SEQ ID NO: 1, extracellular domain 3 (also referred to as EC3 herein) having the amino acid sequence at positions 269 to 383 in the amino acid sequence shown in SEQ ID NO: 1, extracellular domain 4 (also referred to as EC4 herein) having the amino acid sequence at positions 384 to 486 in the amino acid sequence shown in SEQ ID NO: 1, and extracellular domain 5 (also referred to as EC5 herein) having the amino acid sequence at positions 487 to 608 in the amino acid sequence shown in SEQ ID NO: 1. The amino acid sequences of EC1 to EC5 are shown in SEQ ID NOs: 2 to 6, respectively (Table 1).

[0153] 2. Production of anti-CDH6 antibody One example of the anti-CDH6 antibody of the present disclosure may include an anti-CDH6 antibody that recognizes an amino acid sequence containing the amino acid sequence shown in SEQ ID NO: 4 and has internalization activity. One example of the anti-CDH6 antibody of the present disclosure may include an anti-CDH6 antibody that specifically recognizes an amino acid sequence containing the amino acid sequence shown in SEQ ID NO: 4 and has internalization activity. One example of the anti-CDH6 antibody of the present disclosure may include an anti-CDH6 antibody that recognizes an amino acid sequence consisting of the amino acid sequence shown in SEQ ID NO: 4 and has internalization activity. One example of the anti-CDH6 antibody of the present disclosure may include an anti-CDH6 antibody that specifically recognizes an amino acid sequence consisting of the amino acid sequence shown in SEQ ID NO: 4 and has internalization activity. The phrase "specifically recognizes an amino acid sequence containing the amino acid sequence shown in SEQ ID NO: 4" or "specifically recognizes the EC3 domain" applied to the antibody is used to mean that the antibody strongly recognizes or strongly binds to the EC3 domain of CDH6 as compared to other extracellular domains of CDH6.

[0154] The anti-CDH6 antibody of the present disclosure may be derived from any species. Preferred examples of the species may include human, monkey, rat, mouse, and rabbit. When the anti-CDH6 antibody of the present disclosure is derived from a species other than human, it is preferable to humanize or chimerize the anti-CDH6 antibody by well-known techniques. The antibody of the present disclosure may be a polyclonal antibody or a monoclonal antibody, and monoclonal antibodies are preferred.

[0155] The anti-CDH6 antibody of the present disclosure is an antibody that can target tumor cells. Specifically, the anti-CDH6 antibody of the present disclosure has properties such as the ability to recognize tumor cells, the ability to bind to tumor cells, and / or the ability to be internalized into tumor cells by cell uptake. Therefore, the anti-CDH6 antibody of the present disclosure can be conjugated to a compound having antitumor activity via a linker to prepare an antibody-drug conjugate.

[0156] The binding activity of an antibody to tumor cells can be confirmed by flow cytometry. The uptake of an antibody into tumor cells can be confirmed by (1) an assay using a secondary antibody (fluorescently labeled) that binds to the antibody to visualize the antibody taken up by cells under a fluorescence microscope (Cell Death and Differentiation, 2008, 15, 751-761), (2) an assay using a secondary antibody (fluorescently labeled) that binds to the antibody to measure the amount of fluorescence taken up by cells (Molecular Biology of the Cell, Vol. 15, 5268-5282, December 2004), or (3) a Mab-ZAP assay using an immunotoxin that binds to the antibody and releases a toxin to inhibit cell growth when there is cell uptake (Bio Techniques 28:162-165, January 2000). The catalytic region of diphtheria toxin and a recombinant conjugate protein of protein G can be used as an immunotoxin.

[0157] In the present description, the term "high internalization ability" means that the survival rate of CDH6-expressing cells administered with the aforementioned antibody and saporin-labeled anti-rat IgG antibody (shown by the ratio compared to the cell survival rate without antibody addition defined as 100%) is preferably 70% or less, more preferably 60% or less.

[0158] The anti-tumor antibody-drug conjugate of the present disclosure contains a conjugated compound that exerts an anti-tumor effect. Therefore, it is preferable but not essential that the antibody itself has an anti-tumor effect. For the purpose of specifically and / or selectively exerting the cytotoxicity of the anti-tumor compound in tumor cells, it is important and preferable that the antibody has the property of being internalized and moving into tumor cells.

[0159] The anti-CDH6 antibody can be obtained by immunizing an animal with a polypeptide that acts as an antigen by a method commonly practiced in this field, and then recovering and purifying the antibody produced in the organism. It is preferable to use CDH6 that retains its three-dimensional structure as an antigen. Examples of such methods may include DNA immunization.

[0160] The origin of the antigen is not limited to humans, and thus, animals can be immunized with antigens derived from non-human animals such as mice or rats. In this case, antibodies applicable to human diseases can be selected by examining the cross-reactivity of the acquired antibodies that bind to heterologous antigens using human antigens.

[0161] Furthermore, antibody-producing cells that produce antibodies against the antigen can be fused with myeloma cells according to known methods (e.g., Kohler and Milstein, Nature (1975) 256, 495 - 497; and Kennet, R. ed., Monoclonal Antibodies, 365 - 367, Plenum Press, N.Y. (1980)) to establish hybridomas and obtain monoclonal antibodies.

[0162] Hereinafter, methods for obtaining antibodies against CDH6 will be specifically described.

[0163] (1) Preparation of Antigen The antigen can be obtained by causing a host cell to produce a gene encoding the antigen protein according to genetic manipulation. Specifically, a vector capable of expressing the antigen gene is produced, then the vector is introduced into the host cell, whereby the gene is expressed therein, and thereafter, the expressed antigen can be purified. Antibodies can also be obtained by immunizing an animal with antigen-expressing cells based on the above genetic manipulation or a cell line expressing the antigen.

[0164] Alternatively, the antibody can be obtained without using the antigen protein by incorporating the cDNA of the antigen protein into an expression vector, then administering the expression vector to an animal to be immunized, and expressing the antigen protein in the body of the thus-immunized animal, whereby antibodies against the antigen protein are produced therein.

[0165] (2) Production of Anti-CDH6 Monoclonal Antibody The anti-CDH6 antibodies used in the present disclosure are not particularly limited. For example, antibodies specified by the amino acid sequences shown in the sequence listing of the present application can be appropriately used. The anti-CDH6 antibodies used in the present disclosure are preferably antibodies having the following characteristics: (1) Antibodies having the following characteristics: (a) Specifically bind to CDH6, (b) Have the activity of being internalized into CDH6-expressing cells by binding to CDH6; (2) An antibody according to (1) above, wherein CDH6 is human CDH6; or (3) An antibody according to (1) or (2) above, which specifically recognizes EC3 of human CDH6 and has internalization activity.

[0166] The method for obtaining an antibody against CDH6 in the present disclosure is not particularly limited as long as an anti-CDH6 antibody can be obtained. It is preferable to use CDH6 that retains its three-dimensional structure as the antigen.

[0167] One preferred example of a method for obtaining an antibody may include DNA immunization. DNA immunization is a technique involving the steps of transfecting an individual animal (e.g., a mouse or a rat) with an antigen-expressing plasmid, and then inducing immunity against the antigen by expressing the antigen in the individual. Transfection techniques include methods of directly injecting the plasmid into muscle, injecting a transfection reagent such as liposome or polyethyleneimine into a vein, techniques using viral vectors, techniques using a gene gun to inject gold particles attached to the plasmid, hydrodynamic methods of rapidly injecting a large amount of plasmid solution into a vein, etc. Regarding the transfection method of injecting an expression plasmid into muscle, a technique called in vivo electroporation, which involves applying electroporation to the intramuscular injection site of the plasmid, is known as a technique for improving the expression level (Aihara H, Miyazaki J. Nat Biotechnol. September 1998;16(9):867-70 or Mir LM, Bureau MF, Gehl J, Rangara R, Rouy D, Caillaud JM, Delaere P, Branellec D, Schwartz B, Scherman D. Proc Natl Acad Sci U S A. April 13, 1999;96(8):4262-7). This technique can further improve the expression level by treating the muscle with hyaluronidase before intramuscular injection of the plasmid (McMahon JM1, Signori E, Wells KE, Fazio VM, Wells DJ., Gene Ther. August 2001;8(16):1264-70). Furthermore, hybridoma production can be carried out by known methods, and can also be carried out using, for example, the Hybrimune hybridoma production system (Cyto Pulse Sciences, Inc.).

[0168] Specific methods for obtaining monoclonal antibodies may include the following procedures: (a) Incorporate the CDH6 cDNA into an expression vector (e.g., pcDNA3.1; Thermo Fisher Scientific Inc.), and directly administer the vector to an animal to be immunized (e.g., a rat or a mouse) by methods such as electroporation or gene gun, so that CDH6 is expressed in the animal's body, whereby an immune response can be induced. If necessary to enhance the antibody titer, the administration of the vector by electroporation or the like can be carried out once or multiple times, preferably multiple times; (b) Recovery of a tissue (e.g., lymph node) containing antibody-producing cells from the aforementioned animal in which an immune response has been induced; (c) Preparation of myeloma cells (hereinafter referred to as "myeloma") (e.g., mouse myeloma SP2 / 0-ag14 cells); (d) Cell fusion between antibody-producing cells and myeloma; (e) Selection of a group of hybridomas that produce the antibody of interest; (f) Division (cloning) into single cell clones; (g) Optionally, culturing of hybridomas for mass production of monoclonal antibodies, or breeding of animals inoculated with hybridomas; and / or (h) Investigation of the physiological activity (internalization activity) and binding specificity of the monoclonal antibody thus produced, or examination of the properties of the antibody as a labeling reagent.

[0169] Examples of methods for measuring the antibody titer used herein may include, but are not limited to, flow cytometry and Cell-ELISA.

[0170] Examples of hybridoma lines established in this way may include anti-CDH6 antibody-producing hybridomas rG019, rG055, rG056, and rG061. In this description, the antibody produced by anti-CDH6 antibody-producing hybridoma rG019 is referred to as the "rG019 antibody" or simply "rG019", the antibody produced by hybridoma rG055 is referred to as the "rG055 antibody" or simply "rG055", the antibody produced by hybridoma rG056 is referred to as the "rG056 antibody" or simply "rG056", and the antibody produced by hybridoma rG061 is referred to as the "rG061 antibody" or simply "rG061".

[0171] The light chain variable region of the rG019 antibody consists of the amino acid sequence shown in SEQ ID NO: 10. The amino acid sequence of the light chain variable region of the rG019 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 11. The light chain variable region of the rG019 antibody has CDRL1 consisting of the amino acid sequence shown in SEQ ID NO: 12, CDRL2 consisting of the amino acid sequence shown in SEQ ID NO: 13, and CDRL3 consisting of the amino acid sequence shown in SEQ ID NO: 14. The heavy chain variable region of the rG019 antibody consists of the amino acid sequence shown in SEQ ID NO: 15. The amino acid sequence of the heavy chain variable region of the rG019 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 16. The heavy chain variable region of the rG019 antibody has CDRH1 consisting of the amino acid sequence shown in SEQ ID NO: 17, CDRH2 consisting of the amino acid sequence shown in SEQ ID NO: 18, and CDRH3 consisting of the amino acid sequence shown in SEQ ID NO: 19. The sequence of the rG019 antibody is shown in Table 1.

[0172] The light chain variable region of the rG055 antibody consists of the amino acid sequence shown in SEQ ID NO: 20. The amino acid sequence of the light chain variable region of the rG055 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 21. The light chain variable region of the rG055 antibody has CDRL1 consisting of the amino acid sequence shown in SEQ ID NO: 22, CDRL2 consisting of the amino acid sequence shown in SEQ ID NO: 23, and CDRL3 consisting of the amino acid sequence shown in SEQ ID NO: 24. The heavy chain variable region of the rG055 antibody consists of the amino acid sequence shown in SEQ ID NO: 25. The amino acid sequence of the heavy chain variable region of the rG055 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 26. The heavy chain variable region of the rG055 antibody has CDRH1 consisting of the amino acid sequence shown in SEQ ID NO: 27, CDRH2 consisting of the amino acid sequence shown in SEQ ID NO: 28, and CDRH3 consisting of the amino acid sequence shown in SEQ ID NO: 29. The sequence of the rG055 antibody is shown in Table 1.

[0173] The light chain variable region of the rG056 antibody consists of the amino acid sequence shown in SEQ ID NO: 30. The amino acid sequence of the light chain variable region of the rG056 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 31. The light chain variable region of the rG056 antibody has CDRL1 consisting of the amino acid sequence shown in SEQ ID NO: 32, CDRL2 consisting of the amino acid sequence shown in SEQ ID NO: 33, and CDRL3 consisting of the amino acid sequence shown in SEQ ID NO: 34. The heavy chain variable region of the rG056 antibody consists of the amino acid sequence shown in SEQ ID NO: 35. The amino acid sequence of the heavy chain variable region of the rG056 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 36. The heavy chain variable region of the rG056 antibody has CDRH1 consisting of the amino acid sequence shown in SEQ ID NO: 37, CDRH2 consisting of the amino acid sequence shown in SEQ ID NO: 38, and CDRH3 consisting of the amino acid sequence shown in SEQ ID NO: 39. The sequence of the rG056 antibody is shown in Table 1.

[0174] The light chain variable region of the rG061 antibody consists of the amino acid sequence shown in SEQ ID NO: 40. The amino acid sequence of the light chain variable region of the rG061 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 41. The light chain variable region of the rG061 antibody has CDRL1 consisting of the amino acid sequence shown in SEQ ID NO: 42, CDRL2 consisting of the amino acid sequence shown in SEQ ID NO: 43, and CDRL3 consisting of the amino acid sequence shown in SEQ ID NO: 44. The heavy chain variable region of the rG061 antibody consists of the amino acid sequence shown in SEQ ID NO: 45. The amino acid sequence of the heavy chain variable region of the rG061 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 46. The heavy chain variable region of the rG061 antibody has CDRH1 consisting of the amino acid sequence shown in SEQ ID NO: 47, CDRH2 consisting of the amino acid sequence shown in SEQ ID NO: 48, and CDRH3 consisting of the amino acid sequence shown in SEQ ID NO: 49. The sequence of the rG061 antibody is shown in Table 1.

[0175] Furthermore, when steps (a) to (h) in the above “2. Production of anti-CDH6 antibody” are performed again to separately obtain monoclonal antibodies independently, and also when monoclonal antibodies are separately obtained by other methods, antibodies having internalization activity equivalent to that of the rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody can be obtained. One example of such an antibody may include an antibody that binds to the same epitope to which the rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody binds. When a newly prepared monoclonal antibody binds to a partial peptide or partial three-dimensional structure to which the rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody binds, the monoclonal antibody can be determined to bind to the same epitope to which the rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody binds. Furthermore, even if the specific sequence or structure of the epitope is not determined, by confirming that the monoclonal antibody competes with the rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody in the binding of the antibody to CDH6 (i.e., the monoclonal antibody inhibits the binding of the rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody to CDH6), the monoclonal antibody can be determined to bind to the same epitope to which the anti-CDH6 antibody binds. When it is confirmed that the monoclonal antibody binds to the same epitope to which the rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody binds, it is strongly predicted that the monoclonal antibody should have antigen-binding ability, biological activity, and / or internalization activity equivalent to that of the rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody.

[0176] (3) Other antibodies The antibodies of the present disclosure also include chimeric antibodies, humanized antibodies, and human antibodies, as well as genetically engineered antibodies artificially modified for the purpose of reducing heterologous antigenicity to humans, such as the above monoclonal antibodies against CDH6. These antibodies can be produced by known methods.

[0177] Examples of chimeric antibodies can include antibodies in which the variable and constant regions are heterologous to each other, such as chimeric antibodies formed by conjugating the variable regions of antibodies derived from mice or rats to the constant regions derived from humans (see Proc. Natl. Acad. Sci. U.S.A., 81, 6851-6855, (1984)).

[0178] Examples of chimeric antibodies derived from rat anti-human CDH6 antibodies include antibodies consisting of a light chain comprising the light chain variable region of each rat anti-human CDH6 antibody described in this specification (e.g., rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody) and a human-derived constant region, and a heavy chain comprising the heavy chain variable region and a human-derived constant region.

[0179] Other examples of chimeric antibodies derived from rat anti-human CDH6 antibodies include antibodies consisting of a light chain comprising a light chain variable region having substitution of 1 to several residues, 1 to 3 residues, 1 or 2 residues, preferably 1 residue of amino acids in the light chain variable region of each rat anti-human CDH6 antibody described in this specification (e.g., rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody) with other amino acid residues, and a heavy chain comprising a heavy chain variable region having substitution of 1 to several residues, 1 to 3 residues, 1 or 2 residues, preferably 1 residue of amino acids in the heavy chain variable region with other amino acid residues. This antibody can have any given human-derived constant region.

[0180] Other examples of chimeric antibodies derived from rat anti-human CDH6 antibodies include light chains containing a light chain variable region having a substitution of one or two residues, preferably one residue, of amino acids in any one to three CDRs in the light chain variable region of each rat anti-human CDH6 antibody described in this description (e.g., rG019 antibody, rG055 antibody, rG056 antibody, or rG061 antibody) at other amino acid residues, and heavy chains containing a heavy chain variable region having a substitution of one or two residues, preferably one residue, of amino acids in any one to three CDRs in the heavy chain variable region at other amino acid residues. This antibody may have a constant region derived from any given human.

[0181] Examples of chimeric antibodies derived from the rG019 antibody include antibodies consisting of a light chain containing a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 10 and a heavy chain containing a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 15. This antibody may have a constant region derived from any given human.

[0182] Other examples of chimeric antibodies derived from the rG019 antibody include light chains containing a light chain variable region having a substitution of one to several residues, one to three residues, one or two residues, preferably one residue, of amino acids in the light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 10 at other amino acid residues, and heavy chains containing a heavy chain variable region having a substitution of one to several residues, one to three residues, one or two residues, preferably one residue, of amino acids in the heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 15 at other amino acid residues. This antibody may have a constant region derived from any given human.

[0183] Other examples of chimeric antibodies derived from the rG019 antibody include a light chain comprising a variable light chain region having a substitution of one or two residues (preferably one residue) of amino acid in any one to three CDRs in the variable light chain region consisting of the amino acid sequence shown in SEQ ID NO: 10 with other amino acid residues, and a heavy chain comprising a variable heavy chain region having a substitution of one or two residues (preferably one residue) of amino acid in any one to three CDRs in the variable heavy chain region consisting of the amino acid sequence shown in SEQ ID NO: 15 with other amino acid residues. This antibody can have a constant region derived from any given human.

[0184] Other examples of chimeric antibodies derived from the rG019 antibody include an antibody consisting of a light chain comprising a variable light chain region consisting of the amino acid sequence shown in SEQ ID NO: 10 and a heavy chain comprising a variable heavy chain region consisting of the amino acid sequence shown in SEQ ID NO: 58. This antibody can have a constant region derived from any given human. The amino acid sequence shown in SEQ ID NO: 58 is a sequence having a cysteine residue substituted with a proline residue in CDRH2 in the amino acid sequence shown in SEQ ID NO: 15.

[0185] Specific examples of chimeric antibodies derived from the rG019 antibody include an antibody consisting of a light chain consisting of the full-length amino acid sequence of the light chain shown in SEQ ID NO: 53 and a heavy chain consisting of the full-length amino acid sequence of the heavy chain shown in SEQ ID NO: 56. In this description, this chimeric anti-human CDH6 antibody is referred to as "chimeric G019 antibody", "chG019 antibody", or "chG019". The full-length amino acid sequence of the light chain of the chG019 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 54, and the full-length amino acid sequence of the heavy chain of the chG019 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 57.

[0186] The amino acid sequence of the light chain variable region of the chG019 antibody is identical to the amino acid sequence of the light chain variable region of the rG019 antibody and consists of the amino acid sequence shown in SEQ ID NO: 10. The light chain of the chG019 antibody consists of CDRL1, CDRL2, and CDRL3 that are identical to the light chain CDRL1, CDRL2, and CDRL3 of rG019, respectively, and has CDRL1 consisting of the amino acid sequence shown in SEQ ID NO: 12, CDRL2 consisting of the amino acid sequence shown in SEQ ID NO: 13, and CDRL3 consisting of the amino acid sequence shown in SEQ ID NO: 14. The amino acid sequence of the light chain variable region of the chG019 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 55.

[0187] The amino acid sequence of the heavy chain variable region of the chG019 antibody consists of the amino acid sequence shown in SEQ ID NO: 58. The heavy chain of the chG019 antibody has CDRH1 consisting of the amino acid sequence shown in SEQ ID NO: 17, CDRH2 consisting of the amino acid sequence shown in SEQ ID NO: 60, and CDRH3 consisting of the amino acid sequence shown in SEQ ID NO: 19. The amino acid sequence shown in SEQ ID NO: 58 is a sequence having a cysteine residue substituted with a proline residue in CDRH2 of the amino acid sequence shown in SEQ ID NO: 15. CDRH2 consisting of the amino acid sequence shown in SEQ ID NO: 60 is a sequence having a cysteine residue substituted with a proline residue in rG019 CDRH2 shown in SEQ ID NO: 18. The amino acid sequence of the heavy chain variable region of the chG019 antibody is encoded by the nucleotide sequence shown in SEQ ID NO: 59.

[0188] The sequence of the chG019 antibody is shown in Table 1.

[0189] Examples of chimeric antibodies derived from the rat anti-human CDH6 antibody rG055 antibody include chimeric antibodies consisting of a light chain containing a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 20 and a heavy chain containing a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 25. This antibody can have a constant region derived from any given human.

[0190] Examples of chimeric antibodies derived from the rat anti-human CDH6 antibody rG056 antibody include chimeric antibodies consisting of a light chain comprising a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 30 and a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 35. This antibody may have a constant region derived from any given human.

[0191] Examples of chimeric antibodies derived from the rat anti-human CDH6 antibody rG061 antibody include chimeric antibodies consisting of a light chain comprising a light chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 40 and a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence shown in SEQ ID NO: 45. This antibody may have a constant region derived from any given human.

[0192] Examples of humanized antibodies can include antibodies formed by incorporating only complementarity-determining regions (CDRs) into human-derived antibodies (see Nature (1986) 321, pages 522-525), antibodies formed by incorporating amino acid residues derived from CDR sequences as well as some frameworks into human antibodies according to the CDR grafting method (International Publication No. WO90 / 07861), and antibodies formed by modifying the amino acid sequences of some CDRs while maintaining antigen-binding ability.

[0193] In this description, humanized antibodies derived from the rG019 antibody, rG055 antibody, rG056 antibody, rG061 antibody, or chG019 antibody are not limited to specific humanized antibodies as long as they retain all six CDR sequences unique to the rG019 antibody, rG055 antibody, rG056 antibody, rG061 antibody, or chG019 antibody and have internalization activity. The amino acid sequences of some CDRs of this humanized antibody can be further modified as long as it has internalization activity.

[0194] Specific examples of humanized antibodies of the chG019 antibody include: (1) the amino acid sequences shown in SEQ ID NO: 63 or 67; (2) amino acid sequences having at least 95% identity with the above amino acid sequence (1) (preferably, an amino acid sequence having at least 95% sequence identity with the sequence of the framework region except for each CDR sequence); and (3) any one amino acid sequence selected from the group consisting of amino acid sequences containing deletions, substitutions, or additions of one or several amino acids in the above amino acid sequence (1), and a light chain containing a light chain variable region consisting of such an amino acid sequence; (4) the amino acid sequences shown in SEQ ID NO: 71, 75, or 79; (5) amino acid sequences having at least 95% identity with the above amino acid sequence (4) (preferably, an amino acid sequence having at least 95% sequence identity with the sequence of the framework region except for each CDR sequence); and (6) any one amino acid sequence selected from the group consisting of amino acid sequences containing deletions, substitutions, or additions of one or several amino acids in the above amino acid sequence (4), and a heavy chain containing a heavy chain variable region consisting of such an amino acid sequence, and may include any given combination thereof.

[0195] Alternatively, an antibody having a humanized heavy or light chain and other chains derived from a rat antibody or a chimeric antibody can also be used. Examples of such antibodies include: (1) an amino acid sequence shown in SEQ ID NO: 63 or 67; (2) an amino acid sequence having at least 95% identity with the above amino acid sequence (1) (preferably, an amino acid sequence having at least 95% sequence identity with the sequence of the framework region except for each CDR sequence); and (3) any one amino acid sequence selected from the group consisting of amino acid sequences containing deletion, substitution, or addition of one or several amino acids in the above amino acid sequence (1), a light chain comprising a variable region of the light chain; and (4) an amino acid sequence shown in SEQ ID NO: 15, 25, 35, 45, or 58; (5) an amino acid sequence having at least 95% identity with the above amino acid sequence (4) (preferably, an amino acid sequence having at least 95% sequence identity with the sequence of the framework region except for each CDR sequence); and (6) any one amino acid sequence selected from the group consisting of amino acid sequences containing deletion, substitution, or addition of one or several amino acids in the above amino acid sequence (4), a heavy chain comprising a variable region of the heavy chain, and may include any given combination thereof.Other examples of such antibodies include: (1) an amino acid sequence shown in SEQ ID NO: 10, 20, 30, or 40; (2) an amino acid sequence having at least 95% identity with the above amino acid sequence (1) (preferably, an amino acid sequence having at least 95% sequence identity with the sequence of the framework region except for each CDR sequence); and (3) any one selected from the group consisting of amino acid sequences including deletion, substitution, or addition of one or several amino acids in the above amino acid sequence (1), and a light chain containing a variable region of the light chain consisting of such an amino acid sequence; and (4) an amino acid sequence shown in SEQ ID NO: 71, 75, or 79; (5) an amino acid sequence having at least 95% identity with the above amino acid sequence (4) (preferably, an amino acid sequence having at least 95% sequence identity with the sequence of the framework region except for each CDR sequence); and (6) any one selected from the group consisting of amino acid sequences including deletion, substitution, or addition of one or several amino acids in the above amino acid sequence (4), and a heavy chain containing a variable region of the heavy chain consisting of such an amino acid sequence, and may include any given combination thereof.

[0196] Amino acid substitutions in this description are preferably conservative amino acid substitutions. A conservative amino acid substitution is a substitution that occurs within a group of amino acids associated with a particular amino acid side chain. Preferred groups of amino acids are as follows: acidic group = aspartic acid and glutamic acid; basic group = lysine, arginine, and histidine; non-polar group = alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, and tryptophan; and uncharged polar family = glycine, asparagine, glutamine, cysteine, serine, threonine, and tyrosine. Other preferred groups of amino acids are as follows: aliphatic hydroxy group = serine and threonine; amide-containing group = asparagine and glutamine; aliphatic group = alanine, valine, leucine, and isoleucine; and aromatic group = phenylalanine, tryptophan, and tyrosine. Such amino acid substitutions are preferably made without impairing the properties of the substance having the original amino acid sequence.

[0197] Examples of antibodies having the preferred combinations of the above light and heavy chains include antibodies comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 63 (also referred to herein as the hL02 light chain variable region amino acid sequence), or a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 67 (also referred to herein as the hL03 light chain variable region amino acid sequence), and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 71 (also referred to herein as the hH01 heavy chain variable region amino acid sequence), a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 75 (also referred to herein as the hH02 heavy chain variable region amino acid sequence), or a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 79 (also referred to herein as the hH04 heavy chain variable region amino acid sequence). Preferred examples thereof include antibodies comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 63 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 71; antibodies comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 63 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 75; antibodies comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 63 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 79; antibodies comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 67 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 71; antibodies comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 67 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 75; and antibodies comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 67 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 79.More preferred examples include antibodies comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 63 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 71; an antibody comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 63 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 75; an antibody comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 63 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 79; and an antibody comprising a light chain having the light chain variable region amino acid sequence shown in SEQ ID NO: 67 and a heavy chain having the heavy chain variable region amino acid sequence shown in SEQ ID NO: 75.

[0198] Other examples of antibodies having the preferred combinations of the light and heavy chains described above include a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 (also referred to in this description as the hL02 full-length light chain amino acid sequence), or a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 65 (also referred to in this description as the hL03 full-length light chain amino acid sequence), and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 69 (also referred to in this description as the hH01 full-length heavy chain amino acid sequence), a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 73 (also referred to in this description as the hH02 full-length heavy chain amino acid sequence), or a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 77 (also referred to in this description as the hH04 full-length heavy chain amino acid sequence).Preferred examples include antibodies comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 69; antibodies comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 73; antibodies comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 77; antibodies comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 65 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 69; antibodies comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 65 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 73; and antibodies comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 65 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 77.More preferred examples include an antibody comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 69 (in this description, also referred to as "H01L02 antibody" or "H01L02"); an antibody comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 73 (in this description, also referred to as "H02L02 antibody" or "H02L02"); an antibody comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 77 (in this description, also referred to as "H04L02 antibody" or "H04L02"); and an antibody comprising a light chain consisting of the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 65 and a heavy chain consisting of the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 73 (in this description, also referred to as "H02L03 antibody" or "H02L03"). The sequences of the H01L02 antibody, H02L02 antibody, H02L03 antibody, or H04L02 antibody are shown in Table 1.

[0199] By combining together sequences showing high identity with the above heavy chain amino acid sequence and light chain amino acid sequence, it is possible to select antibodies having biological activities equivalent to those of each of the above antibodies. Such identity is generally 80% or more, preferably 90% or more, more preferably 95% or more, and most preferably 99% or more identity. Furthermore, by combining amino acid sequences of the heavy chain and light chain containing substitution, deletion, or addition of one or several amino acid residues with respect to the amino acid sequence of the heavy chain or light chain, it is also possible to select antibodies having biological activities equivalent to those of each of the above antibodies.

[0200] The identity between two types of amino acid sequences can be determined by aligning the sequences using the default parameters of Clustal W version 2 (Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, and Higgins DG (2007), "Clustal W and Clustal X version 2.0", Bioinformatics. 23(21):2947-2948).

[0201] In the full-length amino acid sequence of the hL02 light chain shown in SEQ ID NO: 61, the amino acid sequence consisting of amino acid residues at positions 1 to 20 is a signal sequence, the amino acid sequence consisting of amino acid residues at positions 21 to 128 is a variable region, and it should be noted that the amino acid sequence consisting of amino acid residues at positions 129 to 233 is a constant region. In the full-length nucleotide sequence of the hL02 light chain shown in SEQ ID NO: 62, the nucleotide sequence consisting of nucleotides at positions 1 to 60 encodes a signal sequence, the nucleotide sequence consisting of nucleotides at positions 61 to 384 encodes a variable region, and the nucleotide sequence consisting of nucleotides at positions 385 to 699 encodes a constant region.

[0202] In the full-length amino acid sequence of the hL03 light chain shown in SEQ ID NO: 65, the amino acid sequence consisting of amino acid residues at positions 1 to 20 is a signal sequence, the amino acid sequence consisting of amino acid residues at positions 21 to 128 is a variable region, and the amino acid sequence consisting of amino acid residues at positions 129 to 233 is a constant region. In the full-length nucleotide sequence of the hL03 light chain shown in SEQ ID NO: 66, the nucleotide sequence consisting of nucleotides at positions 1 to 60 encodes a signal sequence, the nucleotide sequence consisting of nucleotides at positions 61 to 384 encodes a variable region, and the nucleotide sequence consisting of nucleotides at positions 385 to 699 encodes a constant region.

[0203] In the full-length amino acid sequence of the hH01 heavy chain shown in SEQ ID NO: 69, the amino acid sequence consisting of amino acid residues at positions 1 to 19 is a signal sequence, the amino acid sequence consisting of amino acid residues at positions 20 to 141 is a variable region, and the amino acid sequence consisting of amino acid residues at positions 142 to 471 is a constant region. In the full-length nucleotide sequence of the hH01 heavy chain shown in SEQ ID NO: 70, the nucleotide sequence consisting of nucleotides at positions 1 to 57 encodes a signal sequence, the nucleotide sequence consisting of nucleotides at positions 58 to 423 encodes a variable region, and the nucleotide sequence consisting of nucleotides at positions 424 to 1413 encodes a constant region.

[0204] In the full-length amino acid sequence of the hH02 heavy chain shown in SEQ ID NO: 73, the amino acid sequence consisting of amino acid residues at positions 1 to 19 is a signal sequence, the amino acid sequence consisting of amino acid residues at positions 20 to 141 is a variable region, and the amino acid sequence consisting of amino acid residues at positions 142 to 471 is a constant region. In the full-length nucleotide sequence of the hH02 heavy chain shown in SEQ ID NO: 74, the nucleotide sequence consisting of nucleotides at positions 1 to 57 encodes a signal sequence, the nucleotide sequence consisting of nucleotides at positions 58 to 423 encodes a variable region, and the nucleotide sequence consisting of nucleotides at positions 424 to 1413 encodes a constant region.

[0205] In the full-length amino acid sequence of the hH04 heavy chain shown in SEQ ID NO: 77, the amino acid sequence consisting of amino acid residues at positions 1 to 19 is a signal sequence, the amino acid sequence consisting of amino acid residues at positions 20 to 141 is a variable region, and the amino acid sequence consisting of amino acid residues at positions 142 to 471 is a constant region. In the full-length nucleotide sequence of the hH04 heavy chain shown in SEQ ID NO: 78, the nucleotide sequence consisting of nucleotides at positions 1 to 57 encodes a signal sequence, the nucleotide sequence consisting of nucleotides at positions 58 to 423 encodes a variable region, and the nucleotide sequence consisting of nucleotides at positions 424 to 1413 encodes a constant region.

[0206]

Table 1

[0207]

Table 2

[0208]

Table 3

[0209]

Table 4

[0210]

Table 5

[0211]

Table 6

[0212]

Table 7

[0213]

Table 8

[0214]

Table 9

[0215]

Table 10

[0216]

Table 11

[0217]

Table 12

[0218]

Table 13

[0219]

Table 14

[0220]

Table 15

[0221]

Table 16

[0222] In this description, Tables 1-1 to 1-16 are collectively also referred to as Table 1.

[0223] Further examples of the antibodies of the present disclosure may include human antibodies that bind to CDH6. An anti-CDH6 human antibody means a human antibody having only the gene sequences of antibodies derived from the human chromosome. An anti-CDH6 human antibody can be obtained by a method using a human antibody-producing mouse having a human chromosome fragment containing the heavy and light chain genes of a human antibody (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, Volume 10, pp. 69-73 (edited by Kitagawa, Y., Matsuda, T., and Iijima, S.), Kluwer Academic Publishers, 1999; Tomizuka, K. et al., Proc. Natl. Acad. Sci. USA (2000) 97, pp. 722-727; etc.).

[0224] The loci of the endogenous immunoglobulin heavy and light chains are disrupted, and then a genetically modified animal is used in which the loci of the human immunoglobulin heavy and light chains are introduced instead using a yeast artificial chromosome (YAC) vector or the like. Then, knockout animals and transgenic animals are produced from such genetically modified animals, and then such animals are mated with each other, whereby such human antibody-producing mice can be specifically produced.

[0225] Alternatively, an anti-CDH6 human antibody can also be obtained by transforming eukaryotic cells according to genetic recombination techniques with cDNA encoding each of the heavy and light chains of such human antibodies, or preferably with a vector containing the cDNA, and then culturing the transformed cells that produce the genetically modified human monoclonal antibody so that the antibody can be obtained from the culture supernatant.

[0226] In this context, eukaryotic cells, and preferably mammalian cells such as CHO cells, lymphocytes, or myelomas, can be used as hosts, for example.

[0227] Furthermore, methods for obtaining phage display-derived human antibodies selected from human antibody libraries (see, e.g., Wormstone, I.M. 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.) are also known.

[0228] For example, a phage display method can be applied that includes the steps of expressing the variable region of a human antibody as a single-chain antibody (scFv) on the surface of a phage, and then selecting phages that bind to an antigen (Nature Biotechnology (2005), 23, (9), pp. 1105-1116).

[0229] By analyzing the phage gene selected because of its ability to bind to an antigen, the DNA sequence encoding the variable region of the human antibody that binds to the antigen can be determined.

[0230] Once the DNA sequence of the scFv that binds to the antigen is determined, an expression vector having the aforementioned sequence can be produced, and the produced expression vector can then be introduced into a suitable host and expressed therein, thereby obtaining a human antibody (International Publication Nos. WO92 / 01047, WO92 / 20791, WO93 / 06213, WO93 / 11236, WO93 / 19172, WO95 / 01438, and WO95 / 15388, Annu. Rev. Immunol (1994) 12, pp. 433-455, Nature Biotechnology (2005) 23(9), pp. 1105-1116).

[0231] If a newly generated human antibody binds to a partial peptide or partial three-dimensional structure to which any one of the rat anti-human CDH6 antibodies, chimeric anti-human CDH6 antibodies, or humanized anti-human CDH6 antibodies (e.g., rG019 antibody, rG055 antibody, rG056 antibody, rG061 antibody, chG019 antibody, H01L02 antibody, H02L02 antibody, H02L03 antibody, or H04L02 antibody) described in this specification binds, the human antibody can be determined to bind to the same epitope to which the rat anti-human CDH6 antibody, chimeric anti-human CDH6 antibody, or humanized anti-human CDH6 antibody binds. Alternatively, by confirming that a human antibody competes with the rat anti-human CDH6 antibody, chimeric anti-human CDH6 antibody, or humanized anti-human CDH6 antibody (e.g., rG019 antibody, rG055 antibody, rG056 antibody, rG061 antibody, chG019 antibody, H01L02 antibody, H02L02 antibody, H02L03 antibody, or H04L02 antibody) described in this specification in the binding of the antibody to CDH6 (e.g., the human antibody inhibits the binding of the rG019 antibody, rG055 antibody, rG056 antibody, rG061 antibody, chG019 antibody, H01L02 antibody, H02L02 antibody, H02L03 antibody, or H04L02 antibody to CDH6, preferably EC3 of CDH6), even if the specific sequence or structure of the epitope has not been determined, the human antibody can be determined to bind to the same epitope to which the rat anti-human CDH6 antibody, chimeric anti-human CDH6 antibody, or humanized anti-human CDH6 antibody described in this specification binds. In this specification, when it is determined by at least one of these determination methods that a human antibody "binds to the same epitope", the newly prepared human antibody is concluded to "bind to the same epitope" as that against the rat anti-human CDH6 antibody, chimeric anti-human CDH6 antibody, or humanized anti-human CDH6 antibody described in this specification.When it is confirmed that a human antibody binds to the same epitope, the human antibody is expected to have biological activity equivalent to that of a rat anti-human CDH6 antibody, a chimeric anti-human CDH6 antibody, or a humanized anti-human CDH6 antibody (e.g., rG019 antibody, rG055 antibody, rG056 antibody, rG061 antibody, chG019 antibody, H01L02 antibody, H02L02 antibody, H02L03 antibody, or H04L02 antibody).

[0232] The chimeric antibody, humanized antibody, or human antibody obtained by the above method is evaluated for their binding activity to the antigen according to known methods or the like, whereby a preferable antibody can be selected.

[0233] One example of another index for comparing antibody characteristics may include antibody stability. A differential scanning calorimeter (DSC) is a device that can rapidly and accurately measure the heat denaturation midpoint (Tm), which serves as a good index for the relative structural stability of a protein. By measuring the Tm value using DSC and comparing the obtained values, the difference in thermal stability can be compared. It is known that the storage stability of an antibody has a certain correlation with the thermal stability of the antibody (Lori Burton et al., Pharmaceutical Development and Technology (2007) 12, pages 265-273), and thus, a preferable antibody can be selected using thermal stability as an index. Other examples of indices for antibody selection may include high yield in a suitable host cell and low aggregation in an aqueous solution. For example, since an antibody with the highest yield does not necessarily exhibit the highest thermal stability, it is necessary to comprehensively determine it based on the aforementioned indices to select the most suitable antibody for administration to humans.

[0234] The antibodies of the present disclosure also include modifications of the antibodies. The modifications are used to mean the antibodies of the present disclosure that are chemically or biologically modified. Examples of such chemical modifications include the attachment of chemical moieties to the amino acid backbone and the chemical modification of N-linked or O-linked hydrocarbon chains. Examples of such biological modifications include antibodies that have undergone post-translational modifications (e.g., N-linked or O-linked glycosylation, N-terminal or C-terminal processing, deamidation, isomerization of aspartic acid, oxidation of methionine, and conversion of N-terminal glutamine or N-terminal glutamate to pyroglutamic acid), and antibodies that have a methionine residue added to their N-terminus as a result of expression using a prokaryotic host cell. In addition, such modifications are also intended to include the antibodies of the present disclosure, or labeled antibodies, such as enzyme-labeled antibodies, fluorescent-labeled antibodies, and affinity-labeled antibodies, for enabling the detection or isolation of the antibodies or antigens. Such modifications of the antibodies of the present disclosure are useful for improving the stability and retention of the antibodies in the blood; reducing antigenicity; detecting or isolating the antibodies or antigens; and the like.

[0235] Furthermore, antibody-dependent cellular cytotoxicity activity can be enhanced by regulating the sugar chain modifications (such as glycosylation, defucosylation, etc.) that bind to the antibodies of the present disclosure. Techniques for regulating the sugar chain modifications of antibodies are known, such as those described in International Publication Nos. WO1999 / 54342, WO2000 / 61739, and WO2002 / 31140, etc., but the techniques are not limited thereto. The antibodies of the present disclosure also include antibodies in terms of the regulated sugar chain modifications described above.

[0236] Once the antibody gene is isolated, an appropriate combination of a host and an expression vector can be used to introduce the gene into an appropriate host to produce an antibody. Specific examples of the antibody gene can be a combination of the gene encoding the heavy chain sequence of the antibody described in this description and the gene encoding the light chain sequence of the antibody described therein. Once the host cell is transformed, such heavy chain sequence gene and light chain sequence gene can be inserted into a single expression vector, or alternatively, these genes can be inserted into different expression vectors, respectively.

[0237] If a eukaryotic cell is used as a host, an animal cell, a plant cell, or a eukaryotic microorganism can be used. In particular, examples of animal cells include COS cells, which are monkey cells (Gluzman, Y., Cell (1981) 23, pp. 175-182, ATCC CRL-1650), mouse fibroblast NIH3T3 (ATCC number CRL-1658), dihydrofolate reductase-deficient cell line of Chinese hamster ovary cells (CHO cells, ATCC CCL-61) (Urlaub, G. and Chasin, L.A. Proc. Natl. Acad. Sci. U.S.A. (1980) 77, pp. 4126-4220), and mammalian cells such as FreeStyle 293F cells (Invitrogen Corp.).

[0238] If a prokaryotic cell is used as a host, for example, Escherichia coli or Bacillus subtilis can be used.

[0239] The antibody gene of interest is introduced into these cells for transformation, and the transformed cells are then cultured in vitro to obtain an antibody. In the above-mentioned culture, the yield may vary depending on the sequence of the antibody. Therefore, it is possible to select an antibody produced as a medicine from antibodies having equivalent binding activity using the yield as an index. Accordingly, the antibody of the present disclosure also includes an antibody obtained by the above method for producing an antibody, which includes the step of culturing the transformed host cell and the step of recovering the antibody of interest or a functional fragment of the antibody from the culture obtained in the above-mentioned step.

[0240] It is known that lysine residues at the carboxyl terminus of the heavy chain of antibodies produced in cultured mammalian cells are deleted (Journal of Chromatography A, 705:129 - 134 (1995)), and that two amino acid residues, glycine and lysine, at the carboxyl terminus of the heavy chain are deleted, and that proline residues newly located at the carboxyl terminus are amidated (Analytical Biochemistry, 360:75 - 83 (2007)). However, such deletions and modifications of these heavy chain sequences have no effect on the antigen - binding activity and effector functions (complement activation, antibody - dependent cell cytotoxicity, etc.) of the antibody. Therefore, the antibodies according to the present disclosure include antibodies and functional fragments of antibodies that have undergone the aforementioned modifications. Specific examples of such antibodies include deletion mutants containing a deletion of one or two amino acids at the carboxyl terminus of the heavy chain, and deletion mutants formed by amidating the aforementioned deletion mutants (for example, heavy chains in which the proline residue at the carboxyl - terminal site is amidated). However, deletion mutants with deletions at the carboxyl terminus of the heavy chain of the antibodies according to the present disclosure are not limited to the above - mentioned deletion mutants as long as they retain antigen - binding activity and effector functions. The two heavy chains constituting the antibodies according to the present disclosure can be any one type of heavy chain selected from the group consisting of full - length antibodies and the above - mentioned deletion mutants, or can be a combination of any two types selected from the aforementioned group. The proportion of each deletion mutant can be affected by the type of cultured mammalian cells producing the antibodies according to the present disclosure and the culture conditions. Examples of the main component of the antibodies according to the present disclosure can include antibodies in which one amino acid residue is deleted at each of the carboxyl termini of the two heavy chains.

[0241] Examples of the isotypes of the antibodies of the present disclosure can include IgG (IgG1, IgG2, IgG3, and IgG4). Among them, IgG1 and IgG4 are preferred.

[0242] Examples of the biological activities of antibodies generally include antigen-binding activity, the activity of being internalized into cells expressing the antigen by binding to the antigen, the activity of neutralizing the activity of the antigen, the activity of enhancing the activity of the antigen, antibody-dependent cell cytotoxicity (ADCC) activity, complement-dependent cell cytotoxicity (CDC) activity, and antibody-dependent cell phagocytosis (ADCP). The function of the antibody according to the present disclosure is binding activity to CDH6, preferably the activity of being internalized into CDH6-expressing cells by binding to CDH6. Furthermore, the antibody of the present disclosure may have ADCC activity, CDC activity, and / or ADCP activity, as well as cell internalization activity.

[0243] The obtained antibody can be purified to a homogeneous state. For the separation and purification of antibodies, separation and purification methods used for ordinary proteins can be used. For example, column chromatography, filtration, ultrafiltration, salting out, dialysis, preparative polyacrylamide gel electrophoresis, and isoelectric focusing can be appropriately selected and combined with each other, whereby the antibody can be separated and purified (Strategies for Protein Purification and Characterization: A Laboratory Course Manual, edited by Daniel R. Marshak et al., Cold Spring Harbor Laboratory Press (1996); and Antibodies: A Laboratory Manual. Ed Harlow and David Lane, Cold Spring Harbor Laboratory (1988)), although examples of separation and purification methods are not limited thereto.

[0244] Examples of chromatography can include affinity chromatography, ion exchange chromatography, hydrophobic chromatography, gel filtration chromatography, reverse phase chromatography, and absorption chromatography.

[0245] These chromatography techniques can be performed using liquid chromatography such as HPLC or FPLC.

[0246] Examples of columns used in affinity chromatography can include Protein A columns and Protein G columns. Examples of columns involving the use of Protein A can include Hyper D, POROS, and Sepharose F.F. (Pharmacia).

[0247] Also, using an antigen-immobilized carrier, an antibody can be purified by utilizing the binding activity of the antibody to the antigen.

[0248] 3. Anti-CDH6 Antibody-Drug Conjugate (1) Drug The anti-CDH6 antibody obtained in the above "2. Production of anti-CDH6 antibody" can be conjugated to a drug via a linker structural moiety to prepare an anti-CDH6 antibody-drug conjugate. The drug is not particularly limited as long as it has a substituent or partial structure that can be connected to the linker structure. The anti-CDH6 antibody-drug conjugate can be used for various purposes according to the conjugated drug. Examples of such drugs can include substances having antitumor activity, substances effective for blood diseases, substances effective for autoimmune diseases, anti-inflammatory substances, anti-microbial substances, anti-fungal substances, anti-parasitic substances, anti-viral substances, and anti-anesthetic substances.

[0249] (1)-1 Antitumor Compound Examples of using an antitumor compound as the compound to be conjugated in the anti-CDH6 antibody-drug conjugate of the present disclosure will be described below. The antitumor compound is not particularly limited as long as the compound has an antitumor effect and has a substituent or partial structure that can be connected to the linker structure. As soon as there is cleavage of part or all of the linker in tumor cells, the antitumor compound moiety is released, whereby the antitumor compound exhibits an antitumor effect. Since the linker is cleaved at the connection position with the drug, the antitumor compound is released in its original structure and exerts its original antitumor effect.

[0250] The anti-CDH6 antibody obtained in the above “2. Production of anti-CDH6 antibody” can be conjugated to an antitumor compound via a linker structure moiety to prepare an anti-CDH6 antibody-drug conjugate.

[0251] As an example of one of the antitumor compounds used in the present disclosure, exatecan, a camptothecin derivative ((1S,9S)-1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo [d] pyrano [3’,4’:6,7] indolizino [1,2-b] quinoline-10,13(9H,15H)-dione) represented by the following formula, can preferably be used.

[0252]

Chemical formula

[0253] The compound can be obtained, for example, by the method described in US Patent Publication No. US2016 / 0297890 or other known methods, and the amino group at the 1-position can preferably be used as the connection position to the linker structure. Furthermore, exatecan can be released in tumor cells, while a part of the linker still adheres to it. However, the compound exhibits an excellent antitumor effect even in such a state.

[0254] Since exatecan has a camptothecin structure, it is known that in an acidic aqueous medium (e.g., around pH 3), the equilibrium shifts to a structure having a formed lactone ring (closed ring), while in a basic aqueous medium (e.g., around pH 10), the equilibrium shifts to a structure having an open lactone ring (open ring). Drug conjugates into which exatecan residues corresponding to such closed-ring and open-ring structures are introduced are also expected to have an equivalent antitumor effect, and it goes without saying that any of such drug conjugates is included within the scope of the present disclosure.

[0255] Other examples of anti-tumor compounds may include anti-tumor compounds described in the literature (Pharmacological Reviews, 68, pp. 3-19, 2016). Specific examples thereof may include auristatins such as doxorubicin, calicheamicin, dolastatin 10, monomethyl auristatin E (MMAE) and monomethyl auristatin F (MMAF), maytansinoids such as DM1 and DM4, pyrrolobenzodiazepine dimer SG2000 (SJG-136), camptothecin derivative SN-38, duocarmycin such as CC-1065, amanitin, daunorubicin, mitomycin C, bleomycin, cytarabine, vincristine, vinblastine, methotrexate, platinum-based anti-tumor agents (cisplatin and its derivatives), and taxol and its derivatives.

[0256] In an antibody-drug conjugate, the number of drug molecules conjugated per antibody molecule is a major factor affecting its efficacy and safety. The production of an antibody-drug conjugate is carried out by specifying reaction conditions such as the amounts of starting materials and reagents used in the reaction to achieve a certain number of conjugated drug molecules. Unlike chemical reactions of low molecular weight compounds, a mixture containing various numbers of conjugated drug molecules is usually obtained. The number of drug molecules conjugated per antibody molecule is defined and shown as an average value, i.e., the average number of conjugated drug molecules. Unless otherwise specified, i.e., except when representing an antibody-drug conjugate having a specific number of conjugated drug molecules included in an antibody-drug conjugate mixture having various numbers of conjugated drug molecules, the number of conjugated drug molecules according to the present disclosure also generally means an average value. The number of exatecan molecules conjugated to an antibody molecule is controllable, and approximately 1 to 10 exatecan molecules can be conjugated as the average number of conjugated drug molecules per antibody. The number of exatecan molecules is preferably 2 to 8, 3 to 8, 4 to 8, 5 to 8, 6 to 8, or 7 to 8, more preferably 5 to 8, still more preferably 7 to 8, and yet even more preferably 8. Those skilled in the art should note that, based on the description of the examples of the present application, they can design a reaction for conjugating the required number of drug molecules to an antibody molecule and obtain an antibody-drug conjugate having a controlled number of conjugated exatecan molecules.

[0257] (2) Linker structure The linker structure for conjugating a drug to an anti-CDH6 antibody in the anti-CDH6 antibody-drug conjugate of the present disclosure will be described.

[0258] In the antibody-drug conjugate of the present application, the linker structure that conjugates the anti-CDH6 antibody to the drug is not particularly limited as long as the resulting antibody-drug conjugate can be used. The linker structure can be appropriately selected and used according to the purpose of use. One example of the linker structure can include linkers described in known literature (Pharmacol Rev 68:3-19, January 2016, Protein Cell DOI 10.1007 / s13238-016-0323-0, etc.). More specific examples thereof can include VC (valine-citrulline), MC (maleimidocaproyl), SMCC (succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate), SPP (N-succinimidyl 4-(2-pyridyldithio)pentanoate), SS (disulfide), SPDB (N-succinimidyl 4-(2-pyridyldithio)butyrate), SS / hydrazone, hydrazone, and carbonate.

[0259] Another example can include the linker structure described in US Patent Publication No. US2016 / 0297890 (for example, those described in paragraphs

[0260] ~

[0289] thereof). Any linker structure given below can preferably be used. It should be noted that the left end of the structure is the connection position to the antibody and the right end is the connection position to the drug. Furthermore, GGFG (SEQ ID NO: 89) in the linker structure given below represents an amino acid sequence consisting of glycine-glycine-phenylalanine-glycine (GGFG) (SEQ ID NO: 89) linked through a peptide bond. -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-, -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH2 CH 2 CH 2 -C(=O)-、 -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-、 -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 -O-CH 2 -C(=O)-、 -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-、 and -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-

[0260] More preferred are the following: -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-、 -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 -O-CH 2 -C(=O)-、および -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-

[0261] Even more preferred are the following: -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-、および -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH2 CH 2 -C(=O)-

[0262] The antibody is at the end opposite to the end where -(Succinimid-3-yl-N) is attached (e.g., in "-(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-」, -CH 2 CH 2 CH 2 CH 2 CH 2 - is connected to the end opposite to the end where the antibody is connected to -(Succinimid-3-yl-N) (left end in the above example), and the anti-tumor compound is connected to the end opposite to the end where the antibody is connected to -(Succinimid-3-yl-N) (in the above example, the CH at the right end 2 -O-CH 2 -C(=O)-'s carbonyl group). "-(Succinimid-3-yl-N)-" has a structure represented by the following formula:

[0263]

Chemical formula

[0264] The 3-position of this partial structure is the connection position to the anti-CDH6 antibody. This connection to the antibody at the 3-position is characterized by forming a thioether bond. The nitrogen atom at the 1-position of this structural moiety is connected to the carbon atom of the methylene present in the linker containing the structure.

[0265] In the antibody-drug conjugate of the present disclosure having exatecan as a drug, a drug-linker structural moiety having any of the structures given below is preferred for conjugation to the antibody. Regarding these drug-linker structural moieties, the average number of conjugated per antibody can be 1 to 10, preferably 2 to 8, more preferably 5 to 8, still more preferably 7 to 8, and even more preferably 8. -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-(NH-DX), -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-(NH-DX), -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-(NH-DX), -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 -O-CH 2 -C(=O)-(NH-DX), -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-(NH-DX), and -(Succinimid-3-yl-N)-CH 2 CH 2-C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-(NH-DX)

[0266] More preferred are the following: -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-(NH-DX), -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 -O-CH 2 -C(=O)-(NH-DX), and -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-(NH-DX)

[0267] Even more preferred are the following: -(Succinimid-3-yl-N)-CH2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-(NH-DX), and -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-(NH-DX)

[0268] -(NH-DX) is the following formula:

[0269] [Chemical formula] It has a structure represented by, which represents a group obtained by removing one hydrogen atom from the amino group at the 1-position of exatecan.

[0270] (3) Method for producing an antibody-drug conjugate The antibody that can be used in the antibody-drug conjugate of the present disclosure is not particularly limited as long as it is an anti-CDH6 antibody having internalization activity or a functional fragment of the antibody, as described in the above section "2. Production of anti-CDH6 antibody" and the examples.

[0271] Next, a typical method for producing the antibody-drug conjugate of the present disclosure will be described. In the following description, it should be noted that the "compound number" shown in each reaction scheme is used to represent the compound. Specifically, each compound is referred to as "compound of formula (1)", "compound (1)", etc. The same applies to other compound numbers.

[0272] (3)-1 Production method 1 An antibody-drug conjugate represented by formula (1) given below, in which an anti-CDH6 antibody is connected to a linker structure via a thioether, can be produced by reacting an antibody having a sulfhydryl group converted from a disulfide bond by reduction of the anti-CDH6 antibody with compound (2), and compound (2) can be obtained by a known method (for example, by the method described in Patent Publication Document US2016 / 297890 (for example, the method described in paragraphs

[0336] to

[0374] )). This antibody-drug conjugate can be produced, for example, by the following method.

[0273] [Number]

[0274] In the formula, AB represents an antibody having a sulfhydryl group. In the formula, L 1 has a structure represented by -(Succinimid-3-yl-N)-, L 1 ’ represents a maleimidyl group represented by the following formula.

[0275] [Chemical formula]

[0276] -L 1 -L X has a structure represented by any of the following formulas: -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-, -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-、 -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-、 -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 -O-CH 2 -C(=O)-、 -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-、および -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-

[0277] Among them, the more preferred ones are as follows: -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-, -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 -O-CH 2 -C(=O)-, and -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-

[0278] Even more preferred are the following: -(Succinimid-3-yl-N)-CH 2 CH 2 CH 2 CH 2 CH 2 -C(=O)-GGFG-NH-CH 2 -O-CH 2 -C(=O)-, and -(Succinimid-3-yl-N)-CH 2 CH 2 -C(=O)-NH-CH 2 CH 2 O-CH 2 CH 2 O-CH 2CH 2 -C(=O)-GGFG-NH-CH 2 CH 2 CH 2 -C(=O)-

[0279] In the above reaction scheme, the antibody-drug conjugate (1) can be understood to have a structure in which one structural moiety from the drug to the linker terminus is connected to one antibody. However, this explanation is provided for convenience, and in fact, there are many cases where a plurality of the aforementioned moieties are connected to one antibody molecule. The same also applies to the description of the production method described below.

[0280] Specifically, the antibody-drug conjugate (1) can be produced by reacting a compound (2) obtainable by a known method (for example, obtainable by the method described in Patent Publication Document US2016 / 297890 (for example, obtainable by the method described in paragraphs

[0336] to

[0374] )) with an antibody (3a) having a sulfhydryl group.

[0281] The antibody (3a) having a sulfhydryl group can be obtained by methods well known to those skilled in the art (Hermanson, G.T., Bioconjugate Techniques, pages 56 - 136, 456 - 493, Academic Press (1996)). Examples of the methods can include, but are not limited to: reacting a Traut reagent with the amino group of the antibody; reacting N-succinimidyl S-acetylthioalkanoate with the amino group of the antibody, followed by a reaction with hydroxylamine; reacting N-succinimidyl 3-(pyridyldithio)propionate with the antibody, followed by a reaction with a reducing agent; reacting the antibody with a reducing agent such as dithiothreitol, 2-mercaptoethanol, or tris(2-carboxyethyl)phosphine hydrochloride (TCEP) to reduce the interchain disulfide bond in the antibody to form a sulfhydryl group.

[0282] Specifically, an antibody having a partially or fully reduced interchain disulfide bond can be obtained by using 0.3 to 3 molar equivalents of TCEP per interchain disulfide bond in the antibody as a reducing agent and reacting the reducing agent with the antibody in a buffer solution containing a chelating agent. Examples of the chelating agent can include ethylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DTPA). The chelating agent can be used at a concentration of 1 mM to 20 mM. Solutions such as sodium phosphate, sodium borate, and sodium acetate can be used as the buffer solution. As a specific example, an antibody (3a) having a partially or fully reduced sulfhydryl group can be obtained by reacting the antibody with TCEP at 4°C to 37°C for 1 to 4 hours.

[0283] It should be noted that by performing an addition reaction of a sulfhydryl group to the drug-linker moiety, the drug-linker moiety can be conjugated by a thioether bond.

[0284] Next, an antibody-drug conjugate (1) in which 2 to 8 drug molecules are conjugated per antibody can be produced using 2 to 20 molar equivalents of compound (2) per antibody having a sulfhydryl group. Specifically, a solution containing compound (2) dissolved therein can be added to a buffer solution containing an antibody (3a) having a sulfhydryl group for the reaction. In this context, a sodium acetate solution, sodium phosphate, sodium borate, etc. can be used as the buffer solution. The pH for the reaction is 5 to 9, and more preferably the reaction can be carried out around pH 7. An organic solvent such as dimethyl sulfoxide (DMSO), dimethylformamide (DMF), dimethylacetamide (DMA), or N-methyl-2-pyrrolidone (NMP) can be used as the solvent for dissolving compound (2). The reaction can be carried out by adding a solution containing compound (2) dissolved in an organic solvent at 1 to 20% v / v to a buffer solution containing an antibody (3a) having a sulfhydryl group. The reaction temperature is 0 to 37°C, more preferably 10 to 25°C, and the reaction time is 0.5 to 2 hours. The reaction can be terminated by inactivating the reactivity of the unreacted compound (2) using a thiol-containing reagent. The thiol-containing reagent is, for example, cysteine or N-acetyl-L-cysteine (NAC). More specifically, the reaction can be terminated by adding 1 to 2 molar equivalents of NAC to the compound (2) used and incubating the obtained mixture at room temperature for 10 to 30 minutes.

[0285] (4) Identification of the antibody-drug conjugate The produced antibody-drug conjugate (1) can be subjected to concentration, buffer exchange, purification, and measurement of the antibody concentration and the average number of conjugated drug molecules per antibody molecule according to the common procedure described below to identify the antibody-drug conjugate (1).

[0286] (4)-1 Common procedure A: Concentration of an aqueous solution of an antibody or an antibody-drug conjugate An antibody or antibody-drug conjugate solution was added to an Amicon Ultra (50,000 MWCO, Millipore Corporation) vessel, and the antibody or antibody-drug conjugate solution was concentrated by centrifugation (centrifugation for 5 to 20 minutes at 2000G to 3800G) using a centrifuge (Allegra X-15R, Beckman Coulter, Inc.).

[0287] (4)-2 Common Procedure B: Measurement of Antibody Concentration Using a UV detector (Nanodrop 1000, Thermo Fisher Scientific Inc.), the antibody concentration was measured according to the method specified by the manufacturer. In this regard, different 280 nm extinction coefficients (1.3 mL mg -1 cm -1 ~1.8 mL mg -1 cm -1 ) were used depending on the antibody.

[0288] (4)-3 Common Procedure C: Buffer Exchange for Antibody A NAP-25 column (catalog number 17-0852-02, GE Healthcare Japan Corporation) using Sephadex G-25 carrier was equilibrated with a phosphate buffer (50 mM, pH 6.0) containing sodium chloride (50 mM) and EDTA (2 mM) (referred to as PBS6.0 / EDTA in this description) according to the method specified by the manufacturer. An aqueous solution of the antibody was applied in an amount of 2.5 mL per NAP-25 column, and then the eluted fraction (3.5 mL) with 3.5 mL of PBS6.0 / EDTA was collected. This fraction was concentrated by Common Procedure A. After measuring the antibody concentration using Common Procedure B, the antibody concentration was adjusted to 20 mg / mL using PBS6.0 / EDTA.

[0289] (4)-4 Common Procedure D: Purification of Antibody-Drug Conjugate The NAP-25 column was equilibrated with any commercially available buffer solution such as acetate buffer (10 mM, pH 5.5; referred to as ABS in this description) containing sorbitol (5%). An aqueous reaction solution of the antibody-drug conjugate (approximately 2.5 mL) was applied to the NAP-25 column, and then elution was performed with a specified amount of buffer solution by the manufacturer to recover the antibody fraction. The recovered fraction was reapplied to the NAP-25 column, and the gel filtration purification process of eluting with the buffer solution was repeated a total of two or three times to obtain an antibody-drug conjugate excluding unconjugated drug linker and low molecular weight compounds (tris(2-carboxyethyl)phosphine hydrochloride (TCEP), N-acetyl-L-cysteine (NAC), and dimethyl sulfoxide).

[0290] (4)-5 Common Procedure E: Measurement of Antibody Concentration in Antibody-Drug Conjugate and Average Number of Conjugated Drug Molecules per Antibody Molecule The conjugated drug concentration in the antibody-drug conjugate can be calculated by measuring the UV absorbance of an aqueous solution of the antibody-drug conjugate at two wavelengths of 280 nm and 370 nm, and then performing the calculations shown below.

[0291] The total absorbance at any given wavelength is equal to the sum of the absorbances of all light-absorbing chemical species present in the system [additivity of absorbance]. Therefore, based on the hypothesis that the molar extinction coefficients of the antibody and the drug do not vary between before and after conjugation between the antibody and the drug, the antibody concentration and the drug concentration in the antibody-drug conjugate are represented by the following equations. A 280 =A D、280 +A A、280 =ε D、280 C D +ε A、280 C A Equation (1) A 370 =A D、370 +A A、370 =ε D、370 C D +ε A、370 CA Equation (2)

[0292] In this context, A 280 represents the absorbance of an aqueous solution of the antibody-drug conjugate at 280 nm, and A 370 represents the absorbance of an aqueous solution of the antibody-drug conjugate at 370 nm, and A A、280 represents the absorbance of the antibody at 280 nm, and A A、370 represents the absorbance of the antibody at 370 nm, and A D、280 represents the absorbance of the conjugate precursor at 280 nm, and A D、370 represents the absorbance of the conjugate precursor at 370 nm, and ε A、280 represents the molar extinction coefficient of the antibody at 280 nm, and ε A、370 represents the molar extinction coefficient of the antibody at 370 nm, and ε D、280 represents the molar extinction coefficient of the conjugate precursor at 280 nm, and ε D、370 represents the molar extinction coefficient of the conjugate precursor at 370 nm, and C A represents the antibody concentration in the antibody-drug conjugate, and C D represents the drug concentration in the antibody-drug conjugate.

[0293] In this context, for ε A、280 , ε A、370 , ε D、280 , and ε D、370 , pre-prepared values (estimated values based on calculations or measurements obtained by UV measurement of the compound) are used. For example, ε A、280 can be estimated from the amino acid sequence of the antibody by a known calculation method (Protein Science, 1995, Vol. 4, 2411-2423). ε A、370 is generally zero. ε D、280 and ε D、370 can be obtained according to Lambert-Beer's law (absorbance = molar concentration × molar extinction coefficient × cell path length) by measuring the absorbance of a solution in which the conjugate precursor used is dissolved at a certain molar concentration. C A and C Dis the A of the aqueous solution of the antibody-drug conjugate 280 and A 370 can be determined by measuring and then solving the simultaneous equations (1) and (2) by substituting these values. Further, by dividing C D by C A the average number of conjugated drug molecules per antibody can be determined.

[0294] (4)-6 Common Procedure F: Measurement of the average number of conjugated drug molecules per antibody molecule in an antibody-drug conjugate-(2) The average number of conjugated drug molecules per antibody molecule in an antibody-drug conjugate can also be determined by high performance liquid chromatography (HPLC) analysis using the following method in addition to the aforementioned "(4)-5 Common Procedure E". Hereinafter, a method for measuring the average number of conjugated drug molecules by HPLC when the antibody is conjugated to the drug linker by a disulfide bond will be described. A person skilled in the art can appropriately measure the average number of conjugated drug molecules by HPLC according to the mode of connection between the antibody and the drug linker with respect to this method.

[0295] F-1. Preparation of a sample for HPLC analysis (reduction of an antibody-drug conjugate) Mix an antibody-drug conjugate solution (approximately 1 mg / mL, 60 μL) with an aqueous solution of dithiothreitol (DTT) (100 mM, 15 μL). By incubating the mixture at 37 °C for 30 minutes, the disulfide bond between the light chain and the heavy chain of the antibody-drug conjugate is cleaved. The resulting sample is used in HPLC analysis.

[0296] F-2. HPLC analysis Perform HPLC analysis under the following measurement conditions.

[0297] HPLC system: Agilent 1290 HPLC system (Agilent Technologies, Inc.) Detector: Ultraviolet absorption spectrometer (measurement wavelength: 280 nm) Column: ACQUITY UPLC BEH Phenyl (2.1×50 mm, 1.7 μm, 130 Å; Waters Corp., P / N 186002884) Column temperature: 80 °C Mobile phase A: Aqueous solution containing 0.10% trifluoroacetic acid (TFA) and 15% 2-propanol Mobile phase B: Acetonitrile solution containing 0.075% TFA and 15% 2-propanol Gradient program: 14% - 36% (0 min - 15 min), 36% - 80% (15 min - 17 min), 80% - 14% (17 min - 17.01 min), and 14% (17.01 min - 25 min) Sample injection: 10 μL

[0298] F-3. Data analysis F-3-1. Compared with unconjugated antibody light (L0) and heavy (H0) chains, the light chain (L i ) bound to the drug molecule and the heavy chain (H i ) bound to the drug molecule exhibit higher hydrophobicity in proportion to the number of conjugated drug molecules and thus have longer retention times. Therefore, these chains elute in the order of, for example, L0 and L1 or H0, H1, H2, and H3. The detection peaks can be assigned to any of L0, L1, H0, H1, H2, and H3 by comparing the retention times with L0 and H0. The number of conjugated drug molecules can be defined by those skilled in the art, but is preferably L0, L1, H0, H1, H2, and H3.

[0299] F-3-2. Since the drug linker has UV absorption, the peak area value is corrected according to the following formula using the molar extinction coefficients of the light or heavy chain and the drug linker, depending on the number of conjugated drug linker molecules.

[0300]

Equation

[0301]

Number

[0302] In this context, values estimated from the amino acid sequences of the light or heavy chains of each antibody can be used as the molar extinction coefficients (280 nm) of the light or heavy chains of the antibody by a known calculation method (Protein Science, 1995, Vol. 4, 2411-2423). In the case of H01L02, molar extinction coefficients of 31710 and 79990 were used as the estimated values for the light and heavy chains, respectively, according to the amino acid sequence of the antibody. The actually measured molar extinction coefficient (280 nm) of a compound in which a maleimide group is converted to a succinimide thioether by the reaction of each drug linker with mercaptoethanol or N-acetylcysteine was used as the molar extinction coefficient (280 nm) of the drug linker. The wavelength for absorbance measurement can be appropriately set by those skilled in the art, but is preferably a wavelength at which the peak of the antibody can be measured, more preferably 280 nm.

[0303] F-3-3. The peak area ratio (%) of each chain is calculated with respect to the total corrected value of the peak areas according to the following formula.

[0304]

Number

[0305] F-3-4. The average number of conjugated drug molecules per antibody molecule in the antibody-drug conjugate is calculated according to the following formula.

[0306] Average number of conjugated drug molecules = (L 0 Peak area ratio × 0 + L 1 Peak area ratio × 1 + H 0 Peak area ratio × 0 + H 1Peak area ratio × 1 + H 2 Peak area ratio × 2 + H 3 (Peak area ratio × 3) / 100 × 2

[0307] It should be noted that in order to ensure the amount of the antibody-drug conjugate, a plurality of antibody-drug conjugates having approximately the same average number (e.g., within ±1 unit) of conjugated drug molecules produced under the same conditions can be mixed to prepare a new lot. In this case, the average number of drug molecules in the new lot falls between the average numbers of drug molecules before mixing.

[0308] One specific example of the antibody-drug conjugate of the present disclosure is the following formula:

[0309]

Chemical formula

[0310] Or the following formula:

[0311]

Chemical formula

[0312] In this context, AB represents the anti-CDH6 antibody disclosed in this description, and the antibody is conjugated to the drug linker via a sulfhydryl group derived from the antibody. In this context, n has the same meaning as that of the so-called DAR (drug-to-antibody ratio) and represents the drug-to-antibody ratio per antibody. Specifically, n represents the number of conjugated drug molecules per antibody molecule, which is a numerical value defined and shown as an average value, that is, the average number of conjugated drug molecules. In the case of the antibody-drug conjugate represented by [Formula 17] or [Formula 18] of the present disclosure, in the measurement by the common procedure F, n can be 2 to 8, preferably 5 to 8, more preferably 7 to 8, and even more preferably 8.

[0313] One example of the antibody-drug conjugate of the present disclosure may include an antibody-drug conjugate having a structure represented by the above formula [Formula 17] or [Formula 18] or a pharmaceutically acceptable salt of the antibody-drug conjugate, wherein the antibody represented by AB includes any one antibody selected from the group consisting of the following antibodies (a) to (g) or a functional fragment of the antibody: (a) An antibody consisting of a light chain comprising the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain comprising the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 69; (b) An antibody consisting of a light chain comprising the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain comprising the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 73; (c) An antibody consisting of a light chain comprising the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 61 and a heavy chain comprising the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 77; (d) An antibody consisting of a light chain comprising the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 65 and a heavy chain comprising the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 69; (e) An antibody consisting of a light chain comprising the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 65 and a heavy chain comprising the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 73; (f) An antibody consisting of a light chain comprising the amino acid sequence at positions 21 to 233 in the full-length light chain amino acid sequence shown in SEQ ID NO: 65 and a heavy chain comprising the amino acid sequence at positions 20 to 471 in the full-length heavy chain amino acid sequence shown in SEQ ID NO: 77; and Any one antibody selected from the group consisting of antibodies (a)-(f), wherein the heavy chain or light chain comprises one or more types of modifications selected from the group consisting of post-translational modifications represented by N-linked glycosylation, O-linked glycosylation, N-terminal processing, C-terminal processing, deamidation, isomerization of aspartic acid, oxidation of methionine, addition of a methionine residue to the N-terminus, amidation of a proline residue, and conversion of an N-terminal glutamine or N-terminal glutamic acid to pyroglutamic acid, and deletion of one or two amino acids at the carboxyl terminus.

[0314] 4. Medicine The anti-CDH6 antibody or functional fragment of the antibody of the present disclosure described in the above section "2. Production of anti-CDH6 antibody" and the Examples binds to CDH6 on the surface of tumor cells and has internalization activity. Therefore, it can be used as a medicine, alone or in combination with additional drugs, particularly for the treatment of cancers such as renal cell tumors or ovarian tumors, such as renal cell carcinoma, clear cell renal cell carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, thyroid cancer, cholangiocarcinoma, lung cancer (e.g., small cell lung cancer or non-small cell lung cancer), glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, or neuroblastoma.

[0315] Furthermore, the anti-CDH6 antibody or functional fragment of the antibody of the present disclosure can be used in the detection of cells expressing CDH6.

[0316] Furthermore, since the anti-CDH6 antibody or functional fragment of the antibody of the present disclosure has internalization activity, it can be applied as an antibody in an antibody-drug conjugate.

[0317] When a drug having antitumor activity such as cytotoxic activity is used as a drug, the anti-CDH6 antibody-drug conjugate of the present disclosure described in the above section "3. Anti-CDH6 antibody-drug conjugate" and in the examples is a conjugate of an anti-CDH6 antibody having internalization activity and / or a functional fragment of the antibody, and a drug having antitumor activity such as cytotoxic activity. Since this anti-CDH6 antibody-drug conjugate exhibits antitumor activity against cancer cells expressing CDH6, it can be used as a medicine, particularly as a therapeutic agent and / or a prophylactic agent for cancer.

[0318] The anti-CDH6 antibody-drug conjugate of the present disclosure may change to a hydrate, for example, by absorbing moisture or having adsorbed water, when it is left in air or subjected to a recrystallization or purification procedure. Such compounds or pharmacologically acceptable salts containing water are also included in the present disclosure.

[0319] When the anti-CDH6 antibody-drug conjugate of the present disclosure has a basic group such as an amino group, it can form a pharmacologically acceptable acid addition salt if desired. Examples of such acid addition salts include hydrohalides such as hydrofluoride, hydrochloride, hydrobromide, and hydroiodide; inorganic acid salts such as nitrate, perchlorate, sulfate, and phosphate; lower alkane sulfonates such as methanesulfonate, trifluoromethanesulfonate, and ethanesulfonate; aryl sulfonates such as benzenesulfonate and p-toluenesulfonate; organic acid salts such as formate, acetate, trifluoroacetate, malate, fumarate, succinate, citrate, tartrate, oxalate, and maleate; and amino acid salts such as ornithine salt, glutamate, and aspartate.

[0320] When the anti-CDH6 antibody-drug conjugate of the present disclosure has an acidic group such as a carboxy group, it may form a pharmaceutically acceptable base addition salt, if necessary. Examples of such base addition salts include alkali metal salts such as sodium salt, potassium salt, and lithium salt; alkaline earth metal salts such as calcium salt and magnesium salt; inorganic salts such as ammonium salt; and organic amine salts such as dibenzylamine salt, morpholine salt, phenylglycine alkyl ester salt, ethylenediamine salt, N-methylglucamine salt, diethylamine salt, triethylamine salt, cyclohexylamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, diethanolamine salt, N-benzyl-N-(2-phenylethoxy)amine salt, piperazine salt, tetramethylammonium salt, and tris(hydroxymethyl)aminomethane salt.

[0321] The present disclosure may also include an anti-CDH6 antibody-drug conjugate in which one or more atoms constituting the antibody-drug conjugate are replaced with isotopes of the atoms. There are two types of isotopes: radioactive isotopes and stable isotopes. Examples of isotopes may include isotopes of hydrogen (2H and 3H), isotopes of carbon (11C, 13C, and 14C), isotopes of nitrogen (13N and 15N), isotopes of oxygen (15O, 17O, and 18O), and isotopes of fluorine (18F). Compositions containing antibody-drug conjugates labeled with such isotopes are useful, for example, as therapeutic agents, prophylactic agents, research reagents, assay reagents, diagnostic agents, and imaging agents for in vivo diagnosis. All antibody-drug conjugates labeled with isotopes, and mixtures of antibody-drug conjugates labeled with isotopes in any given ratio, are included in the present disclosure. Antibody-drug conjugates labeled with isotopes can be produced, for example, by using isotope-labeled starting materials in place of the starting materials for the production methods of the present disclosure mentioned later, according to methods known in the art.

[0322] In vitro cytotoxicity can be measured, for example, based on the activity of suppressing the cell proliferation response. For example, a cancer cell line overexpressing CDH6 is cultured, and an anti-CDH6 antibody-drug conjugate is added to the culture system at various concentrations. Then, its inhibitory activity against lesion formation, colony formation, and spheroid growth can be measured. In this context, for example, by using a cancer cell line derived from a renal cell tumor or an ovarian tumor, the cell growth inhibitory activity against a renal cell tumor or an ovarian tumor can be examined.

[0323] The in vivo therapeutic effect against cancer in experimental animals can be measured, for example, by administering an anti-CDH6 antibody-drug conjugate to nude mice inoculated with a tumor cell line highly expressing CDH6 and then measuring the changes in the cancer cells. In this context, for example, by using an animal model derived from immunodeficient mice by inoculating cells derived from renal cell carcinoma, clear cell renal cell carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, or thyroid cancer, the therapeutic effect against renal cell carcinoma, clear cell renal cell carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, or thyroid cancer can be measured.

[0324] The types of cancer to which the anti-CDH6 antibody-drug conjugate of the present disclosure is applicable are not particularly limited as long as the cancer expresses CDH6 in the cancer cells to be treated. Examples thereof include renal cell carcinoma (e.g., clear cell renal carcinoma or papillary renal cell carcinoma), ovarian cancer, ovarian serous adenocarcinoma, thyroid cancer, cholangiocarcinoma, lung cancer (e.g., small cell lung cancer or non-small cell lung cancer), glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, and neuroblastoma. However, as long as the cancer expresses CDH6, the cancer is not limited thereto. Examples thereof include renal cell carcinoma, clear cell renal carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, clear cell carcinoma of the ovary, endometrioid carcinoma of the ovary, mucinous tumor of the ovary, thyroid cancer, cholangiocarcinoma, lung cancer, non-small cell lung cancer, cervical cancer, brain tumor, head and neck cancer, sarcoma, osteosarcoma, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, neuroblastoma, colorectal cancer, gastric cancer, endometrial cancer, nasopharyngeal cancer, prostate cancer, or cancer associated with von Hippel-Lindau disease. Preferred examples thereof include renal cell carcinoma, clear cell renal carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, clear cell carcinoma of the ovary, endometrioid carcinoma of the ovary, mucinous tumor of the ovary, thyroid cancer, cholangiocarcinoma, lung cancer, non-small cell lung cancer, cervical cancer, brain tumor, head and neck cancer, sarcoma, osteosarcoma, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, and neuroblastoma. More preferred examples of the cancer may include renal cell carcinoma (e.g., clear cell renal carcinoma and papillary renal cell carcinoma) and ovarian cancer. Even more preferred examples of the cancer may include ovarian cancer (e.g., epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer). The cancer may be metastatic.

[0325] The anti-CDH6 antibody-drug conjugate of the present disclosure can preferably be administered to mammals, more preferably to humans.

[0326] Pharmaceutical Compositions and Modes of Administration The substances used in the pharmaceutical composition containing the anti-CDH6 antibody-drug conjugate of the present disclosure can be appropriately selected from pharmaceutical additives and other commonly used ones in this field from the viewpoint of the applied dosage or the applied concentration, and then can be used.

[0327] The anti-CDH6 antibody-drug conjugates of the present disclosure can be administered as a pharmaceutical composition comprising one or more pharmaceutically compatible components. For example, pharmaceutical compositions typically include one or more pharmaceutical carriers (e.g., sterile liquids (e.g., water and oils (petroleum, and oils of animal, vegetable, or synthetic origin (e.g., peanut oil, soybean oil, mineral oil, and sesame oil)))). When the pharmaceutical composition is administered intravenously, water is a more typical carrier. Aqueous saline solutions, aqueous dextrose solutions, and aqueous glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical vehicles are known in the art. Optionally, the composition can also include trace amounts of humectants, emulsifiers, or pH buffering agents. Examples of suitable pharmaceutical carriers are disclosed in "Remington’s Pharmaceutical Sciences" by E.W. Martin. The formulation corresponds to the mode of administration.

[0328] In some embodiments, the antibody-drug conjugate (ADC) comprises Formula 3. In some embodiments, the ADC comprising Formula 3 is administered to a patient exhibiting resistance to platinum-based cancer therapy. In some embodiments, the ADC comprising Formula 3 is administered to a platinum-resistant cancer patient having cancer recurrence within 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after completion of platinum-based cancer therapy. In some embodiments, the ADC comprising Formula 3 is administered to a platinum-resistant cancer patient having disease recurrence within at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after completion of platinum-based cancer therapy. In some embodiments, the ADC comprising Formula 3 is administered to a platinum-resistant cancer patient having disease recurrence within at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after completion of a carboplatin and / or paclitaxel regimen.

[0329] In some embodiments, the ADC comprising Formula 3 is administered after treatment with platinum / taxane chemotherapy, such as carboplatin / docetaxel, cisplatin / paclitaxel, and carboplatin / paclitaxel / bevacizumab regimens.

[0330] In some embodiments, the ADC comprising Formula 3 is administered after treatment with one or more of the following platinum-based chemotherapies, such as carboplatin / liposomal doxorubicin, carboplatin / gemcitabine, cisplatin / gemcitabine, carboplatin / ifosfamide, cisplatin / ifosfamide, oxaliplatin / 5-FU / leucovorin, and oxaliplatin / capecitabine regimens.

[0331] In some embodiments, the ADC comprising Formula 3 is preferably administered to patients in need thereof for the treatment of platinum-resistant disease recurrence, having ovarian cancer, non-small cell lung cancer (NSCLC), breast cancer, bladder cancer, endometrial cancer, castration-resistant prostate cancer (CRPC), and other cancers expressing CDH6. In some embodiments, ovarian cancer includes epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

[0332] In some embodiments, the ADC comprising Formula 3 is indicated for the treatment of patients with ovarian cancer in which the disease has progressed or recurred after platinum-based chemotherapy. In some embodiments, the ADC comprising Formula 3 is indicated for the treatment of advanced ovarian cancer in women who have failed platinum-based primary chemotherapy regimens. In some embodiments, the ADC comprising Formula 3 is indicated for the treatment of ovarian cancer after disease progression following chemotherapy.

[0333] In some embodiments, the ADC comprising Formula 3 is indicated for the treatment of patients with metastatic ovarian cancer as a single agent, at the time of first or subsequent chemotherapy or after disease progression thereafter. In some embodiments, the ADC comprising Formula 3 is indicated for the treatment of patients with metastatic cancer of the ovary after failure of primary or subsequent therapy.

[0334] Various delivery systems are known and can be used to administer the anti-CDH6 antibody-drug conjugates of the present disclosure. Examples of the administration routes can include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, and subcutaneous routes. Administration can be effected, for example, by injection or bolus injection. According to a specific preferred embodiment, the administration of the above antibody-drug conjugate is effected by injection. Parenteral administration is the preferred route of administration.

[0335] According to a representative embodiment, the pharmaceutical composition is formulated as a pharmaceutical composition suitable for intravenous administration to humans according to conventional procedures. The composition for intravenous administration is typically a solution in the form of a sterile and isotonic aqueous buffer solution. Optionally, the pharmaceutical can also contain solubilizing agents and local anesthetics (e.g., lidocaine) to relieve pain in the injection area. Generally, the above components are provided together, separately or in the form of a mixture in a unit dosage form, as a lyophilized powder or anhydrous concentrate contained in a container obtained, for example, by sealing in an ampoule or sachet indicating the amount of the active agent. If the pharmaceutical is to be administered by injection, it can be administered, for example, using an injection bottle containing sterile pharmaceutical-grade water or physiological saline. If the pharmaceutical is to be administered by injection, an ampoule of sterile water for injection or physiological saline can be provided such that the above components are admixed with each other prior to administration.

[0336] The pharmaceutical composition of the present disclosure can be a pharmaceutical composition containing only the anti-CDH6 antibody-drug conjugate of the present application, or can be a pharmaceutical composition containing an anti-CDH6 antibody-drug conjugate and at least one other therapeutic agent for cancer. The anti-CDH6 antibody-drug conjugate of the present disclosure can also be administered together with an additional therapeutic agent for cancer, thereby enhancing the anti-cancer effect. Such additional anti-cancer agents can be administered to an individual together with, simultaneously with, separately from, or sequentially with the antibody-drug conjugate. Alternatively, the additional anti-cancer agent and the anti-CDH6 antibody-drug conjugate can each be administered to the subject at different dosing intervals. In the present disclosure, the phrase "second drug" means a therapeutic agent other than the anti-CDH6 antibody-drug conjugate of the present disclosure. The additional anti-cancer agent can be the "second drug". However, in the present disclosure, the "second drug" does not necessarily have to be a drug used in so-called "second-line therapy". Examples of such therapeutic agents or second drugs for cancer, or additional anti-cancer agents, include tyrosine kinase inhibitors including imatinib, sunitinib, and regorafenib, CDK4 / 6 inhibitors including palbociclib, HSP90 inhibitors including TAS-116, MEK inhibitors including MEK162, and immune checkpoint inhibitors including nivolumab, pembrolizumab, and ipilimumab. However, the therapeutic agent for cancer is not limited thereto as long as the drug has anti-tumor activity. If the cancer is resistant to a specific anti-cancer agent, the second drug can be selected from anti-cancer agents other than the specific therapeutic agent.

[0337] Such pharmaceutical compositions can be prepared as a lyophilized formulation or in the form of a liquid formulation, as a formulation having a selected composition and the required purity. A pharmaceutical composition prepared as a lyophilized formulation can be a formulation containing suitable pharmaceutical additives used in the art. Similarly, the liquid formulation can be prepared such that it contains various pharmaceutical additives used in the art.

[0338] The composition and concentration of the pharmaceutical composition also vary depending on the administration method. Regarding the affinity of the anti-CDH6 antibody-drug conjugate contained in the pharmaceutical composition of the present disclosure for the antigen, that is, the dissociation constant (Kd value) of the anti-CDH6 antibody-drug conjugate for the antigen, as the affinity increases (i.e., the Kd value is low), the pharmaceutical composition can exhibit drug efficacy even if its application dose decreases. Therefore, the application dose of the antibody-drug conjugate can also be determined by setting the application dose based on the state of the affinity of the antibody-drug conjugate for the antigen. When the antibody-drug conjugate of the present disclosure is administered to a human, it can be administered, for example, once or multiple times at intervals of 1 to 180 days at a dose of approximately 0.001 to 100 mg / kg.

[0339] Methods of treatment and uses The present disclosure provides a method for treating cancer, comprising the step of administering the anti-CDH6 antibody-drug conjugate disclosed herein. Any of the disclosed anti-CDH6 ADCs for use in treating cancer are also provided herein.

[0340] In some embodiments, the cancer is a CDH6-expressing cancer. CDH6-expressing cancers can include, but are not limited to, renal cell carcinoma, clear cell renal carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, thyroid cancer, cholangiocarcinoma, lung cancer, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms tumor, and neuroblastoma.

[0341] For the purposes of the present disclosure, the term "CDH6-overexpressing cancer" is not particularly limited insofar as it is recognized as a CDH6-overexpressing cancer by those skilled in the art. Preferred examples of CDH6-overexpressing cancers may include cancers that are scored highly for CDH6 expression in immunohistochemistry (IHC) or in situ hybridization (ISH) or next-generation sequencing (NGS). The in situ hybridization methods of the present disclosure include fluorescence in situ hybridization (FISH) and dual-color in situ hybridization (DISH).

[0342] A method for scoring the degree of CDH6 expression by immunohistochemistry, or a method for determining positive or negative for CDH6 expression by in situ hybridization, is not particularly limited insofar as it is recognized by those skilled in the art.

[0343] In some embodiments, the ADCs, therapies, and uses of the present disclosure can also be used as pharmaceutical compositions for the treatment of cancer, comprising an antibody-drug conjugate, a salt thereof, or a hydrate thereof, used in the present disclosure as an active ingredient, and a pharmaceutically acceptable formulation ingredient.

[0344] In some embodiments, the ADCs, therapies, and uses of the present disclosure exhibit excellent antitumor activity against cancers that are resistant to existing anticancer drugs (i.e., resistant cancers), particularly cancers that have acquired resistance to existing anticancer drugs (i.e., secondary resistant cancers). Thus, the ADCs for the treatment of the present disclosure, when applied to a patient population having cancers resistant to existing anticancer drugs (patients with a treatment history using existing anticancer drugs) among cancer patients, exhibit significant antitumor effects. In particular, the cancers to be treated are resistant or refractory to treatment with platinum-based chemotherapeutic drugs (i.e., cisplatin, carboplatin), chemotherapeutic drugs (i.e., docetaxel, gemcitabine, liposomal doxorubicin, paclitaxel, topotecan), PARP inhibitor therapy (i.e., niraparib, olaparib, rucaparib, talazoparib), immune checkpoint inhibitor therapy (i.e., nivolumab, pembrolizumab, atezolizumab, avelumab, ipilimumab, durvalumab, tislelizumab, sintilimab, semaprilimab), angiogenesis inhibitor therapy (i.e., bevacizumab, everolimus), and / or VEGFR-TKI therapy (i.e., pazopanib, sunitinib, sorafenib, cabozantinib, lenvatinib, axitinib).

[0345] The ADCs, therapies, and uses of the present disclosure can preferably be used for the treatment of inoperable or recurrent cancers.

[0346] The ADCs, methods, or uses for the treatment of the present disclosure can delay the development of cancer cells, inhibit their growth, and further kill cancer cells. These effects can enable cancer patients to be free from the symptoms caused by cancer or achieve an improvement in the quality of life (QOL) of cancer patients, and the therapeutic effect can be realized by supporting the lives of cancer patients. Even if the anti-CDH6 antibody-drug conjugate of the present disclosure does not succeed in killing cancer cells, it can provide a higher QOL for cancer patients by inhibiting or controlling the growth of cancer cells, and at the same time achieve a longer survival period.

[0347] In some embodiments of the method and use, a subject having cancer (e.g., CDH6-expressing cancer) can be administered from about 0.1 to about 15 mg / kg, from about 0.5 to about 12 mg / kg, from about 1.0 to about 10 mg / kg, or from about 4 to about 8 mg / kg. In some embodiments, the dosage is from about 1.0 mg / kg to about 12 mg / kg, from about 1.6 mg / kg to about 3.2 mg / kg, from about 1.6 mg / kg to about 4.8 mg / kg, from about 1.6 mg / kg to about 5.4 mg / kg, from about 1.6 mg / kg to about 5.6 mg / kg, from about 1.6 mg / kg to about 6.4 mg / kg, from about 1.6 mg / kg to about 8.0 mg / kg, from about 1.6 mg / kg to about 9.6 mg / kg, from about 3.2 mg / kg to about 4.8 mg / kg, from about 3.2 mg / kg to about 5.4 mg / kg, from about 3.2 mg / kg to about 5.6 mg / kg, from about 3.2 mg / kg to about 6.4 mg / kg, from about 3.2 mg / kg to about 8.0 mg / kg, from about 3.2 mg / kg to about 9.6 mg / kg, from about 4.8 mg / kg to about 5.4 mg / kg, from about 4.8 mg / kg to about 5.6 mg / kg, from about 4.8 mg / kg to about 6.4 mg / kg, from about 4.8 mg / kg to about 8.0 mg / kg, from about 4.8 mg / kg to about 9.6 mg / kg, from about 5.4 mg / kg to about 5.6 mg / kg, from about 5.4 mg / kg to about 6.4 mg / kg, from about 5.4 mg / kg to about 8.0 mg / kg, from about 5.4 mg / kg to about 9.6 mg / kg, from about 5.6 mg / kg to about 6.4 mg / kg, from about 5.6 mg / kg to about 8.0 mg / kg, from about 5.6 mg / kg to about 9.6 mg / kg, from about 6.4 mg / kg to about 9.6 mg / kg, from about 6.4 mg / kg to about 8.0 mg / kg, or from about 8.0 mg / kg to about 9.6 mg / kg.In some embodiments, the dose of the ADC administered to the subject can be about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, about 8.2, about 8.3, about 8.4, about 8.5, about 8.6, about 8.7, about 8.8, about 8.9, about 9.0, about 9.1, about 9.2, about 9.3, about 9.4, about 9.5, about 9.6, about 9.7, about 9.8, about 9.9, about 10.0, about 10.1, about 10.2, about 10.3, about 10.4, about 10.5, about 10.6, about 10.7, about 10.8, about 10.9, about 11.0, about 11.1, about 11.2, about 11.3, about 11.4, about 11.5, about 11.6, about 11.7, about 11.8, about 11.9, or about 12.0 mg / kg, or an amount greater than that. In a preferred embodiment, the dose is about 1.6 mg / kg, about 3.2 mg / kg, about 4.8 mg / kg, about 5.4 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg, about 8.0 mg / kg, or about 9.6 mg / kg, but more preferably about 3.2 mg / kg, about 4.8 mg / kg, about 5.4 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg, about 8.0 mg / kg, or about 9.6 mg / kg, but even more preferably about 4.8 mg / kg, about 5.4 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg, or about 8.0 mg / kg, but even more preferably about 4.8 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg, or about 8.0 mg / kg.

[0348] In a preferred embodiment, the dosage is about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 4.0 mg / kg, about 4.1 mg / kg, about 4.2 mg / kg, about 4.3 mg / kg, about 4.4 mg / kg, about 4.5 mg / kg, about 4.6 mg / kg, about 4.7 mg / kg, about 4.8 mg / kg, about 4.9 mg / kg, about 5.0 mg / kg, about 5.1 mg / kg, about 5.2 mg / kg, about 5.3 mg / kg, about 5.4 mg / kg, about 5.5 mg / kg, about 5.6 mg / kg, about 5.7 mg / kg, about 5.8 mg / kg, about 5.9 mg / kg, about 6.0 mg / kg, about 6.1 mg / kg, about 6.2 mg / kg, about 6.3 mg / kg, or about 6.4 mg / kg, but more preferably about 4.8 mg / kg, about 4.9 mg / kg, about 5.0 mg / kg, about 5.1 mg / kg, about 5.2 mg / kg, about 5.3 mg / kg, about 5.4 mg / kg, about 5.5 mg / kg, about 5.6 mg / kg, about 5.7 mg / kg, about 5.8 mg / kg, about 5.9 mg / kg, about 6.0 mg / kg, about 6.1 mg / kg, about 6.2 mg / kg, about 6.3 mg / kg, or about 6.4 mg / kg.

[0349] In some embodiments of the methods and uses, a subject having cancer (e.g., CDH6-expressing cancer) may be administered 0.1 to 15 mg / kg, 0.5 to 12 mg / kg, 1.0 to 10 mg / kg, or 4 to 8 mg / kg. In some embodiments, the dosage may be 1.0 mg / kg to 12 mg / kg, 1.6 mg / kg to 3.2 mg / kg, 1.6 mg / kg to 4.8 mg / kg, 1.6 mg / kg to 5.4 mg / kg, 1.6 mg / kg to 5.6 mg / kg, 1.6 mg / kg to 6.4 mg / kg, 1.6 mg / kg to 8.0 mg / kg, 1.6 mg / kg to 9.6 mg / kg, 3.2 mg / kg to 4.8 mg / kg, 3.2 mg / kg to 5.4 mg / kg, 3.2 mg / kg to 5.6 mg / kg, 3.2 mg / kg to 6.4 mg / kg, 3.2 mg / kg to 8.0 mg / kg, 3.2 mg / kg to 9.6 mg / kg, 4.8 mg / kg to 5.4 mg / kg, 4.8 mg / kg to 5.6 mg / kg, 4.8 mg / kg to 6.4 mg / kg, 4.8 mg / kg to 8.0 mg / kg, 4.8 mg / kg to 9.6 mg / kg, 5.4 mg / kg to 5.6 mg / kg, 5.4 mg / kg to 6.4 mg / kg, 5.4 mg / kg to 8.0 mg / kg, 5.4 mg / kg to 9.6 mg / kg, 5.6 mg / kg to 6.4 mg / kg, 5.6 mg / kg to 8.0 mg / kg, 5.6 mg / kg to 9.6 mg / kg, 6.4 mg / kg to 9.6 mg / kg, 6.4 mg / kg to 8.0 mg / kg, or 8.0 mg / kg to 9.6 mg / kg.In some embodiments, the dose of the ADC administered to the subject can be 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, or 12.0 mg / kg, or an amount greater than that. In preferred embodiments, the dose can be 1.6 mg / kg, 3.2 mg / kg, 4.8 mg / kg, 5.4 mg / kg, 5.6 mg / kg, 6.4 mg / kg, 8.0 mg / kg, 9.6 mg / kg, but more preferably 3.2 mg / kg, 4.8 mg / kg, 5.4 mg / kg, 5.6 mg / kg, 6.4 mg / kg, 8.0 mg / kg, or 9.6 mg / kg, but even more preferably 4.8 mg / kg, 5.4 mg / kg, 5.6 mg / kg, 6.4 mg / kg, or 8.0 mg / kg, but even more preferably 4.8 mg / kg, 5.6 mg / kg, 6.4 mg / kg, or 8.0 mg / kg.

[0350] In a preferred embodiment, the dosage may be 3.2 mg / kg, 3.3 mg / kg, 3.4 mg / kg, 3.5 mg / kg, 3.6 mg / kg, 3.7 mg / kg, 3.8 mg / kg, 3.9 mg / kg, 4.0 mg / kg, 4.1 mg / kg, 4.2 mg / kg, 4.3 mg / kg, 4.4 mg / kg, 4.5 mg / kg, 4.6 mg / kg, 4.7 mg / kg, 4.8 mg / kg, 4.9 mg / kg, 5.0 mg / kg, 5.1 mg / kg, 5.2 mg / kg, 5.3 mg / kg, 5.4 mg / kg, 5.5 mg / kg, 5.6 mg / kg, 5.7 mg / kg, 5.8 mg / kg, 5.9 mg / kg, 6.0 mg / kg, 6.1 mg / kg, 6.2 mg / kg, 6.3 mg / kg, or 6.4 mg / kg, but more preferably 4.8 mg / kg, 4.9 mg / kg, 5.0 mg / kg, 5.1 mg / kg, 5.2 mg / kg, 5.3 mg / kg, 5.4 mg / kg, 5.5 mg / kg, 5.6 mg / kg, 5.7 mg / kg, 5.8 mg / kg, 5.9 mg / kg, 6.0 mg / kg, 6.1 mg / kg, 6.2 mg / kg, 6.3 mg / kg, or 6.4 mg / kg.

[0351] In some embodiments of the method and use, a subject having cancer (e.g., CDH6-expressing cancer) is administered from about 5 to 3000 mg, from about 5 to 2500 mg, from about 5 to 2000 mg, from about 5 to about 1500 mg, from about 5 to about 1000 mg, from about 10 to about 3000 mg, from about 10 to about 2500 mg, from about 10 to about 2000 mg, from about 10 to about 1500 mg, from about 10 to about 1000 mg, from about 25 to about 2500 mg, from about 25 to about 2000 mg, from about 25 to about 1500 mg, from about 25 to about 1000 mg, from about 50 to about 2500 mg, from about 50 to about 2000 mg, from about 50 to about 1500 mg, from about 50 to about 1000 mg, from about 50 to about 950 mg, from about 50 to about 900 mg, from about 50 to about 850 mg, from about 50 to about 800 mg, from about 50 to about 750 mg, from about 50 to about 700 mg, from about 50 to about 650 mg, from about 50 to about 600 mg, from about 50 to about 550 mg, from about 50 to about 500 mg, from about 50 to about 450 mg, from about 50 to about 400 mg, from about 50 to about 350 mg, from about 50 to about 300 mg, from about 50 to about 250 mg, from about 100 to about 2500 mg, from about 100 to about 2000 mg, from about 100 to about 1500 mg, from about 100 to about 1000 mg, from about 100 to about 950 mg, from about 100 to about 900 mg, from about 100 to about 850 mg, from about 100 to about 800 mg, from about 100 to about 750 mg, from about 100 to about 700 mg, from about 100 to about 650 mg, from about 100 to about 600 mg, from about 100 to about 550 mg, from about 100 to about 500 mg, from about 100 to about 450 mg, from about 100 to about 400 mg, from about 100 to about 350 mg, from about 100 to about 300 mg, from about 100 to about 250 mg, from about 150 to about 2000 mg, from about 150 to about 1500 mg, from about 150 to about 1000 mg, from about 150 to about 950 mg, from about 150 to about 900 mg, from about 150 to about 850 mg, from about 150 to about 800 mg, from about 150 to about 750 mg, from about 150 to about 700 mg, from about 150 to about 650 mg, from about 150 to about 600 mg, from about 150 to about 550 mg, from about 150 to about 500 mg, from about 150 to about 450 mg, from about 150 to about 400 mg, from about 150 to about 350 mg, from about 150 to about 300 mg, from about 150 to about 250 mg, from about 200 to about 1000 mg, from about 200 to about 950 mg, from about 200 to about 900 mg, from about 200 to about 850 mg, from about 200 to about 800 mg, from about 200 to about 750 mg, from about 200 to about 700 mg, from about 200 to about 650 mg, from about 200 to about 600 mg, from about 200 to about 550 mg, from about 200 to about 500 mgfrom about 200 to about 450 mg, from about 200 to about 400 mg, from about 200 to about 350 mg, from about 200 to about 300 mg, from about 200 to about 250 mg, from about 250 to about 1000 mg, from about 250 to about 950 mg, from about 250 to about 900 mg, from about 250 to about 850 mg, from about 250 to about 800 mg, from about 250 to about 750 mg, from about 250 to about 700 mg, from about 250 to about 650 mg, from about 250 to about 600 mg, from about 250 to about 550 mg, from about 250 to about 500 mg, from about 250 to about 450 mg, from about 250 to about 400 mg, from about 250 to about 350 mg, from about 250 to about 300 mg, from about 300 to about 1000 mg, from about 300 to about 950 mg, from about 300 to about 900 mg, from about 300 to about 850 mg, from about 300 to about 800 mg, from about 300 to about 750 mg, from about 300 to about 700 mg, from about 300 to about 650 mg, from about 300 to about 600 mg, from about 300 to about 550 mg, from about 300 to about 500 mg, from about 300 to about 450 mg, from about 300 to about 400 mg, from about 300 to about 350 mg, from about 350 to about 1000 mg, from about 350 to about 950 mg, from about 350 to about 900 mg, from about 350 to about 850 mg, from about 350 to about 800 mg, from about 350 to about 750 mg, from about 350 to about 700 mg, from about 350 to about 650 mg, from about 350 to about 600 mg, from about 350 to about 550 mg, from about 350 to about 500 mg, from about 350 to about 450 mg, from about 350 to about 400 mg, from about 400 to about 1000 mg, from about 400 to about 950 mg, from about 400 to about 900 mg, from about 400 to about 850 mg, from about 400 to about 800 mg, from about 400 to about 750 mg, from about 400 to about 700 mg, from about 400 to about 650 mg, from about 400 to about 600 mg, from about 400 to about 550 mg, from about 400 to about 500 mg, from about 400 to about 450 mg, from about 450 to about 1000 mg, from about 450 to about 950 mg, from about 450 to about 900 mg, from about 450 to about 850 mg, from about 450 to about 800 mg, from about 450 to about 750 mg, from about 450 to about 700 mg, from about 450 to about 650 mg, from about 450 to about 600 mg, from about 450 to about 550 mg, from about 450 to about 500 mg, from about 500 to about 1000 mg, from about 500 to about 950 mg, from about 500 to about 900 mg, from about 500 to about 850 mg, from about 500 to about 800 mg, from about 500 to about 750 mg, from about 500 to about 700 mg, from about 500 to about 650 mg, from about 500 to about 600 mg, or from about 500 to about 550 mg may be administered. In other words, in some embodiments,The dose of the ADC to be administered to the subject is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 205 mg, about 210 mg, about 215 mg, about 220 mg, about 225 mg, about 230 mg, about 235 mg, about 240 mg, about 245 mg, about 250 mg, about 255 mg, about 260 mg, about 265 mg, about 270 mg, about 275 mg, about 280 mg, about 285 mg, about 290 mg, about 295 mg, about 300 mg, about 305 mg, about 310 mg, about 315 mg, about 320 mg, about 325 mg, about 330 mg, about 335 mg, about 340 mg, about 345 mg, about 350 mg, about 355 mg, about 360 mg, about 365 mg, about 370 mg, about 375 mg, about 380 mg, about 385 mg, about 390 mg, about 395 mg, about 400 mg, about 405 mg, about 410 mg, about 415 mg, about 420 mg, about 425 mg, about 430 mg, about 435 mg, about 440 mg, about 445 mg, about 450 mg, about 455 mg, about 460 mg, about 465 mg, about 470 mg, about 475 mg, about 480 mg, about 485 mg, about 490 mg, about 495 mg, about 500 mg, about 505 mg, about 510 mg, about 515 mg, about 520 mg, about 525 mg, about 530 mg, about 535 mg, about 540 mg, about 545 mg, about 550 mg, about 555 mg, about 560 mg, about 565 mg, about 570 mg, about 575 mg, about 580 mg, about 585 mg, about 590 mg, about 595 mg, about 600 mg, about 605 mg, about 610 mg, about 615 mg, about 620 mg, about 625 mg, about 630 mg, about 635 mg, about 640 mg, about 645 mg, about 650 mg, about 655 mg, about 660 mg, about 665 mg, about 670 mg, about 675 mg, about 680 mg, about 685 mg, about 690 mg, about 695 mg, about 700 mg, about 705 mg, about 710 mg, about 715 mg, about 720 mg, about 725 mg, about 730 mg, about 735 mg, about 740 mg, about 745 mg, about 750 mg, about 755 mg, about 760 mg, about 765 mg, about 770 mg, about 775 mg, about 780 mg, about 785 mg, about 790 mg, about 795 mg,about 800 mg, about 805 mg, about 810 mg, about 815 mg, about 820 mg, about 825 mg, about 830 mg, about 835 mg, about 840 mg, about 845 mg, about 850 mg, about 855 mg, about 860 mg, about 865 mg, about 870 mg, about 875 mg, about 880 mg, about 885 mg, about 890 mg, about 895 mg, about 900 mg, about 905 mg, about 910 mg, about 915 mg, about 920 mg, about 925 mg, about 930 mg, about 935 mg, about 940 mg, about 945 mg, about 950 mg, about 955 mg, about 960 mg, about 965 mg, about 970 mg, about 975 mg, about 980 mg, about 985 mg, about 990 mg, about 995 mg, about 1000 mg, about 1025 mg, about 1050 mg, about 1075 mg, about 1100 mg, about 1125 mg, about 1150 mg, about 1175 mg, about 1200 mg, about 1225 mg, about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about 1350 mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450 mg, about 1475 mg, about 1500 mg, about 1525 mg, about 1550 mg, about 1575 mg, about 1600 mg, about 1625 mg, about 1650 mg, about 1675 mg, about 1700 mg, about 1725 mg, about 1750 mg, about 1775 mg, about 1800 mg, about 1825 mg, about 1850 mg, about 1875 mg, about 1900 mg, about 1925 mg, about 1950 mg, about 1975 mg, or about 2000 mg, or any value therebetween.,

[0352] In a preferred embodiment, the dose of the ADC administered to the subject is about 200 mg, about 205 mg, about 210 mg, about 215 mg, about 220 mg, about 225 mg, about 230 mg, about 235 mg, about 240 mg, about 245 mg, about 250 mg, about 255 mg, about 260 mg, about 265 mg, about 270 mg, about 275 mg, about 280 mg, about 285 mg, about 290 mg, about 295 mg, about 300 mg, about 305 mg, about 310 mg, about 315 mg, about 320 mg, about 325 mg, about 330 mg, about 335 mg, about 340 mg, about 345 mg, about 350 mg, about 355 mg, about 360 mg, about 365 mg, about 370 mg, about 375 mg, about 380 mg, about 385 mg, about 390 mg, about 395 mg, about 400 mg, about 405 mg, about 410 mg, about 415 mg, about 420 mg, about 425 mg, about 430 mg, about 435 mg, about 440 mg, about 445 mg, about 450 mg, about 455 mg, about 460 mg, about 465 mg, about 470 mg, about 475 mg, about 480 mg, about 485 mg, about 490 mg, about 495 mg, about 500 mg, about 505 mg, about 510 mg, about 515 mg, about 520 mg, about 525 mg, about 530 mg, about 535 mg, about 540 mg, about 545 mg, about 550 mg, about 555 mg, about 560 mg, about 565 mg, about 570 mg, about 575 mg, about 580 mg, about 585 mg, about 590 mg, about 595 mg, about 600 mg, about 605 mg, about 610 mg, about 615 mg, about 620 mg, about 625 mg, about 630 mg, about 635 mg, about 640 mg, about 645 mg, about 650 mg, about 655 mg, about 660 mg, about 665 mg, about 670 mg, about 675 mg, about 680 mg, about 685 mg, about 690 mg, about 695 mg, about 700 mg, about 705 mg, about 710 mg, about 715 mg, about 720 mg, about 725 mg, about 730 mg, about 735 mg, about 740 mg, about 745 mg, about 750 mg, about 755 mg, about 760 mg, about 765 mg, about 770 mg, about 775 mg, about 780 mg, about 785 mg, about 790 mg, about 795 mg, about 800 mg, about 805 mg, about 810 mg, about 815 mg, about 820 mg, about 825 mg, about 830 mg, about 835 mg, about 840 mg, about 845 mg, about 850 mg, about 855 mg, about 860 mg, about 865 mg, about 870 mg, about 875 mg, about 880 mg, about 885 mg,It can be about 890 mg, about 895 mg, about 900 mg, about 905 mg, about 910 mg, about 915 mg, about 920 mg, about 925 mg, about 930 mg, about 935 mg, about 940 mg, about 945 mg, about 950 mg, about 955 mg, about 960 mg, about 965 mg, about 970 mg, about 975 mg, about 980 mg, about 985 mg, about 990 mg, about 995 mg, or about 1000 mg.

[0353] In some embodiments of the method and use, a subject having cancer (e.g., CDH6-expressing cancer) is administered 5 to 3000 mg, 5 to 2500 mg, 5 to 2000 mg, 5 to 1500 mg, 5 to 1000 mg, 10 to 3000 mg, 10 to 2500 mg, 10 to 2000 mg, 10 to 1500 mg, 10 to 1000 mg, 25 to 2500 mg, 25 to 2000 mg, 25 to 1500 mg, 25 to 1000 mg, 50 to 2500 mg, 50 to 2000 mg, 50 to 1500 mg, 50 to 1000 mg, 50 to 950 mg, 50 to 900 mg, 50 to 850 mg, 50 to 800 mg, 50 to 750 mg, 50 to 700 mg, 50 to 650 mg, 50 to 600 mg, 50 to 550 mg, 50 to 500 mg, 50 to 450 mg, 50 to 400 mg, 50 to 350 mg, 50 to 300 mg, 50 to 250 mg, 100 to 2500 mg, 100 to 2000 mg, 100 to 1500 mg, 100 to 1000 mg, 100 to 950 mg, 100 to 900 mg, 100 to 850 mg, 100 to 800 mg, 100 to 750 mg, 100 to 700 mg, 100 to 650 mg, 100 to 600 mg, 100 to 550 mg, 100 to 500 mg, 100 to 450 mg, 100 to 400 mg, 100 to 350 mg, 100 to 300 mg, 100 to 250 mg, 150 to 2000 mg, 150 to 1500 mg, 150 to 1000 mg, 150 to 950 mg, 150 to 900 mg, 150 to 850 mg, 150 to 800 mg, 150 to 750 mg, 150 to 700 mg, 150 to 650 mg, 150 to 600 mg, 150 to 550 mg, 150 to 500 mg, 150 to 450 mg, 150 to 400 mg, 150 to 350 mg, 150 to 300 mg, 150 to 250 mg, 200 to 1000 mg, 200 to 950 mg, 200 to 900 mg, 200 to 850 mg, 200 to 800 mg, 200 to 750 mg, 200 to 700 mg, 200 to 650 mg, 200 to 600 mg, 200 to 550 mg, 200 to 500 mg, 200 to 450 mg, 200 to 400 mg, 200 to 350 mg, 200 to 300 mg, 200 to 250 mg, 250 to 1000 mg, 250 to 950 mg, 250 to 900 mg, 250 to 850 mg, 250 to 800 mg, 250 to 750 mg, 250 to 700 mg, 250 to 650 mg, 250 to 600 mg, 250 to 550 mg,250 - 500 mg, 250 - 450 mg, 250 - 400 mg, 250 - 350 mg, 250 - 300 mg, 300 - 1000 mg, 300 - 950 mg, 300 - 900 mg, 300 - 850 mg, 300 - 800 mg, 300 - 750 mg, 300 - 700 mg, 300 - 650 mg, 300 - 600 mg, 300 - 550 mg, 300 - 500 mg, 300 - 450 mg, 300 - 400 mg, 300 - 350 mg, 350 - 1000 mg, 350 - 950 mg, 350 - 900 mg, 350 - 850 mg, 350 - 800 mg, 350 - 750 mg, 350 - 700 mg, 350 - 650 mg, 350 - 600 mg, 350 - 550 mg, 350 - 500 mg, 350 - 450 mg, 350 - 400 mg, 400 - 1000 mg, 400 - 950 mg, 400 - 900 mg, 400 - 850 mg, 400 - 800 mg, 400 - 750 mg, 400 - 700 mg, 400 - 650 mg, 400 - 600 mg, 400 - 550 mg, 400 - 500 mg, 400 - 450 mg, 450 - 1000 mg, 450 - 950 mg, 450 - 900 mg, 450 - 850 mg, 450 - 800 mg, 450 - 750 mg, 450 - 700 mg, 450 - 650 mg, 450 - 600 mg, 450 - 550 mg, 450 - 500 mg, 500 - 1000 mg, 500 - 950 mg, 500 - 900 mg, 500 - 850 mg, 500 - 800 mg, 500 - 750 mg, 500 - 700 mg, 500 - 650 mg, 500 - 600 mg, or 500 - 550 mg may be administered. In other words, in some embodiments, the dose of the ADC administered to the subject is 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg, 295 mg, 300 mg, 305 mg, 310 mg, 315 mg, 320 mg, 325 mg, 330 mg, 335 mg, 340 mg, 345 mg,350 mg, 355 mg, 360 mg, 365 mg, 370 mg, 375 mg, 380 mg, 385 mg, 390 mg, 395 mg, 400 mg, 405 mg, 410 mg, 415 mg, 420 mg, 425 mg, 430 mg, 435 mg, 440 mg, 445 mg, 450 mg, 455 mg, 460 mg, 465 mg, 470 mg, 475 mg, 480 mg, 485 mg, 490 mg, 495 mg, 500 mg, 505 mg, 510 mg, 515 mg, 520 mg, 525 mg, 530 mg, 535 mg, 540 mg, 545 mg, 550 mg, 555 mg, 560 mg, 565 mg, 570 mg, 575 mg, 580 mg, 585 mg, 590 mg, 595 mg, 600 mg, 605 mg, 610 mg, 615 mg, 620 mg, 625 mg, 630 mg, 635 mg, 640 mg, 645 mg, 650 mg, 655 mg, 660 mg, 665 mg, 670 mg, 675 mg, 680 mg, 685 mg, 690 mg, 695 mg, 700 mg, 705 mg, 710 mg, 715 mg, 720 mg, 725 mg, 730 mg, 735 mg, 740 mg, 745 mg, 750 mg, 755 mg, 760 mg, 765 mg, 770 mg, 775 mg, 780 mg, 785 mg, 790 mg, 795 mg, 800 mg, 805 mg, 810 mg, 815 mg, 820 mg, 825 mg, 830 mg, 835 mg, 840 mg, 845 mg, 850 mg, 855 mg, 860 mg, 865 mg, 870 mg, 875 mg, 880 mg, 885 mg, 890 mg, 895 mg, 900 mg, 905 mg, 910 mg, 915 mg, 920 mg, 925 mg, 930 mg, 935 mg, 940 mg, 945 mg, 950 mg, 955 mg, 960 mg, 965 mg, 970 mg, 975 mg, 980 mg, 985 mg, 990 mg, 995 mg, 1000 mg, 1025 mg, 1050 mg, 1075 mg, 1100 mg, 1125 mg, 1150 mg, 1175 mg, 1200 mg, 1225 mg, 1250 mg, 1275 mg, 1300 mg, 1325 mg, 1350 mg, 1375 mg, 1400 mg, 1425 mg, 1450 mg, 1475 mg, 1500 mg, 1525 mg, 1550 mg, 1575 mg, 1600 mg, 1625 mg, 1650 mg, 1675 mg, 1700 mg, 1725 mg, 1750 mgIt can be 1775 mg, 1800 mg, 1825 mg, 1850 mg, 1875 mg, 1900 mg, 1925 mg, 1950 mg, 1975 mg, or 2000 mg, or any value in between.

[0354] In a preferred embodiment, the dose of the ADC administered to the subject is 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg, 295 mg, 300 mg, 305 mg, 310 mg, 315 mg, 320 mg, 325 mg, 330 mg, 335 mg, 340 mg, 345 mg, 350 mg, 355 mg, 360 mg, 365 mg, 370 mg, 375 mg, 380 mg, 385 mg, 390 mg, 395 mg, 400 mg, 405 mg, 410 mg, 415 mg, 420 mg, 425 mg, 430 mg, 435 mg, 440 mg, 445 mg, 450 mg, 455 mg, 460 mg, 465 mg, 470 mg, 475 mg, 480 mg, 485 mg, 490 mg, 495 mg, 500 mg, 505 mg, 510 mg, 515 mg, 520 mg, 525 mg, 530 mg, 535 mg, 540 mg, 545 mg, 550 mg, 555 mg, 560 mg, 565 mg, 570 mg, 575 mg, 580 mg, 585 mg, 590 mg, 595 mg, 600 mg, 605 mg, 610 mg, 615 mg, 620 mg, 625 mg, 630 mg, 635 mg, 640 mg, 645 mg, 650 mg, 655 mg, 660 mg, 665 mg, 670 mg, 675 mg, 680 mg, 685 mg, 690 mg, 695 mg, 700 mg, 705 mg, 710 mg, 715 mg, 720 mg, 725 mg, 730 mg, 735 mg, 740 mg, 745 mg, 750 mg, 755 mg, 760 mg, 765 mg, 770 mg, 775 mg, 780 mg, 785 mg, 790 mg, 795 mg, 800 mg, 805 mg, 810 mg, 815 mg, 820 mg, 825 mg, 830 mg, 835 mg, 840 mg, 845 mg, 850 mg, 855 mg, 860 mg, 865 mg, 870 mg, 875 mg, 880 mg, 885 mg, 890 mg, 895 mg, 900 mg, 905 mg, 910 mg, 915 mg, 920 mg, 925 mg, 930 mg, 935 mg, 940 mg, 945 mg, 950 mg, 955 mg, 960 mg, 965 mg, 970 mg, 975 mg, 980 mg, 985 mg, 990 mg, 995 mg,Or it can be 1000 mg.,

[0355] In some embodiments of the methods and uses, the anti-CDH6 ADC or its pharmaceutical composition is administered to a subject having cancer via parenteral administration. Preferred parenteral routes of administration include, but are not limited to, injections such as intravenous, intramuscular, and subcutaneous injections. The anti-CDH6 antibody-drug conjugate used in the present disclosure can be expected to exert a therapeutic effect by application as a systemic therapy to a patient and additionally by local application to cancer tissue.

[0356] The timing or regimen of administration can be once every week (q1w), once every two weeks (q2w), once every three weeks (q3w), once every four weeks (q4w), once every five weeks (q5w), once every six weeks (q6w), once every seven weeks (q7w), once every eight weeks (q8w), once every nine weeks (q9w), or once every ten weeks (q10w), but is preferably once every three or four weeks, and more preferably once every three weeks.

[0357] The dosing regimen can be adjusted to provide an optimal desired therapeutic effect (e.g., a therapeutic effect such as tumor regression or remission). The dosing below is shown once every three weeks, but can also be provided at q1w, q2w, q4w, q5w, q6w, q7w, q8w, q9w, or q10w. For example, in some embodiments, the dosing regimen is 0.1 mg / kg once every three weeks (q3w), 0.2 mg / kg once every three weeks (q3w), 0.3 mg / kg once every three weeks (q3w), 0.4 mg / kg once every three weeks (q3w), 0.5 mg / kg once every three weeks (q3w), 0.6 mg / kg once every three weeks (q3w), 0.7 mg / kg once every three weeks (q3w), 0.8 mg / kg once every three weeks (q3w), 0.9 mg / kg once every three weeks (q3w), 1.0 mg / kg once every three weeks (q3w), 1.1 mg / kg once every three weeks (q3w), 1.2 mg / kg once every three weeks (q3w), 1.3 mg / kg once every three weeks (q3w), 1.4 mg / kg once every three weeks (q3w), 1.5 mg / kg once every three weeks (q3w), 1.6 mg / kg once every three weeks (q3w), 1.7 mg / kg once every three weeks (q3w), 1.8 mg / kg once every three weeks (q3w), 1.9 mg / kg once every three weeks (q3w), 2.0 mg / kg once every three weeks (q3w), 2.1 mg / kg once every three weeks (q3w), 2.2 mg / kg once every three weeks (q3w), 2.3 mg / kg once every three weeks (q3w), 2.4 mg / kg once every three weeks (q3w), 2.5 mg / kg once every three weeks (q3w), 2.6 mg / kg once every three weeks (q3w), 2.7 mg / kg once every three weeks (q3w), 2.8 mg / kg once every three weeks (q3w), 2.9 mg / kg once every three weeks (q3w), 3.0 mg / kg once every three weeks (q3w), 3.1 mg / kg once every three weeks (q3w), 3.2 mg / kg once every three weeks (q3w), 3.3 mg / kg once every three weeks (q3w), 3.4 mg / kg once every three weeks (q3w), 3.5 mg / kg once every three weeks (q3w), 3.6 mg / kg once every three weeks (q3w), 3.7 mg / kg once every three weeks (q3w), 3.8 mg / kg once every three weeks (q3w), 3.9 mg / kg once every three weeks (q3w), 4.0 mg / kg once every three weeks (q3w), 4.1 mg / kg once every three weeks (q3w), 4.2 mg / kg once every three weeks (q3w), 4.3 mg / kg once every three weeks (q3w), 4.4 mg / kg once every three weeks (q3w), 4.5 mg / kg once every three weeks (q3w), 4.6 mg / kg once every three weeks (q3w), 4.7 mg / kg once every three weeks (q3w), 4.8 mg / kg once every three weeks (q3w), 4.9 mg / kg once every three weeks (q3w), 5.0 mg / kg once every three weeks (q3w), 5.1 mg / kg once every three weeks (q3w), 5.2 mg / kg once every three weeks (q3w), 5.3 mg / kg once every three weeks (q3w), 5.4 mg / kg once every three weeks (q3w), 5.5 mg / kg once every three weeks (q3w), 5.6 mg / kg once every three weeks (q3w), 5.7 mg / kg once every three weeks (q3w), 5.8 mg / kg once every three weeks (q3w), 5.9 mg / kg once every three weeks (q3w), 6.0 mg / kg once every three weeks (q3w), 6.1 mg / kg once every three weeks (q3w), 6.2 mg / kg once every three weeks (q3w), 6.3 mg / kg once every three weeks (q3w), 6.4 mg / kg once every three weeks (q3w), 6.5 mg / kg once every three weeks (q3w), 6.6 mg / kg once every three weeks (q3w), 6.7 mg / kg once every three weeks (q3w), 6.8 mg / kg once every three weeks (q3w), 6.9 mg / kg once every three weeks (q3w), 7.0 mg / kg once every three weeks (q3w), 7.1 mg / kg once every three weeks (q3w), 7.2 mg / kg once every three weeks (q3w), 7.3 mg / kg once every three weeks (q3w), 7.4 mg / kg once every three weeks (q3w), 7.5 mg / kg once every three weeks (q3w), 7.6 mg / kg once every three weeks (q3w), 7.7 mg / kg once every three weeks (q3w), 7.8 mg / kg once every three weeks (q3w), 7.9 mg / kg once every three weeks (q3w), 8.0 mg / kg once every three weeks (q3w), 8.1 mg / kg once every three weeks (q3w), 8.2 mg / kg once every three weeks (q3w), 8.3 mg / kg once every three weeks (q3w), 8.4 mg / kg once every three weeks (q3w), 8.5 mg / kg once every three weeks (q3w), 8.6 mg / kg once every three weeks (q3w), 8.7 mg / kg once every three weeks (q3w), 8.8 mg / kg once every three weeks (q3w), 8.9 mg / kg once every three weeks (q3w), 9.0 mg / kg once every three weeks (q3w), 9.1 mg / kg once every three weeks (q3w), 9.2 mg / kg once every three weeks (q3w), 9.3 mg / kg once every three weeks (q3w), 9.4 mg / kg once every three weeks (q3w), 9.5 mg / kg once every three weeks (q3w), 9.6 mg / kg once every three weeks (q3w), 9.7 mg / kg once every three weeks (q3w), 9.8 mg / kg once every three weeks (q3w), 9.9 mg / kg once every three weeks (q3w), 10.0 mg / kg once every three weeks (q3w), 10.1 mg / kg once every three weeks (q3w), 10.2 mg / kg once every three weeks (q3w), 10.3 mg / kg once every three weeks (q3w), 10.4 mg / kg once every three weeks (q3w), 10.5 mg / kg once every three weeks (q3w), 10.6 mg / kg once every three weeks (q3w), 10.7 mg / kg once every three weeks (q3w), 10.8 mg / kg once every three weeks (q3w), 10.9 mg / kg once every three weeks (q3w), 11.0 mg / kg once every three weeks (q3w), 11.1 mg / kg once every three weeks (q3w), 11.2 mg / kg once every three weeks (q3w), 11.3 mg / kg once every three weeks (q3w), 11.4 mg / kg once every three weeks (q3w), 11.5 mg / kg once every three weeks (q3w), 11.6 mg / kg once every three weeks (q3w), 11.7 mg / kg once every three weeks (q3w), 11.8 mg / kg once every three weeks (q3w), 11.9 mg / kg once every three weeks (q3w), or 12.0 mg / kg once every three weeks (q3w). In a preferred embodiment, the dosing regimen is 1.6 mg / kg once every three weeks (q3w), 3.2 mg / kg once every three weeks (q3w), 4.8 mg / kg once every three weeks (q3w), 6.4 mg / kg once every three weeks (q3w), 8.0 mg / kg once every three weeks (q3w), 9.6 mg / kg once every three weeks (q3w), but more preferably 3.2 mg / kg once every three weeks (q3w), 4.8 mg / kg once every three weeks (q3w), 6.4 mg / kg once every three weeks (q3w), 8.0 mg / kg once every three weeks (q3w), or 9.6 mg / kg once every three weeks (q3w), but even more preferably 6.4 mg / kg once every three weeks (q3w) or 8.0 mg / kg once every three weeks (q3w), but even more preferably 8.0 mg / kg once every three weeks (q3w) may be used.

[0358] In a preferred embodiment, the dosing regimen is 3.2 mg / kg once every three weeks (q3w), 3.3 mg / kg once every three weeks (q3w), 3.4 mg / kg once every three weeks (q3w), 3.5 mg / kg once every three weeks (q3w), 3.6 mg / kg once every three weeks (q3w), 3.7 mg / kg once every three weeks (q3w), 3.8 mg / kg once every three weeks (q3w), 3.9 mg / kg once every three weeks (q3w), 4.0 mg / kg once every three weeks (q3w), 4.1 mg / kg once every three weeks (q3w), 4.2 mg / kg once every three weeks (q3w), 4.3 mg / kg once every three weeks (q3w), 4.4 mg / kg once every three weeks (q3w), 4.5 mg / kg once every three weeks (q3w), 4.6 mg / kg once every three weeks (q3w), 4.7 mg / kg once every three weeks (q3w), 4.8 mg / kg once every three weeks (q3w), 4.9 mg / kg once every three weeks (q3w), 5.0 mg / kg once every three weeks (q3w), 5.1 mg / kg once every three weeks (q3w), 5.2 mg / kg once every three weeks (q3w), 5.3 mg / kg once every three weeks (q3w), 5.4 mg / kg once every three weeks (q3w), 5.5 mg / kg once every three weeks (q3w), 5.6 mg / kg once every three weeks (q3w), 5.7 mg / kg once every three weeks (q3w), 5.8 mg / kg once every three weeks (q3w), 5.9 mg / kg once every three weeks (q3w), 6.0 mg / kg once every three weeks (q3w), 6.1 mg / kg once every three weeks (q3w), 6.2 mg / kg once every three weeks (q3w), 6.3 mg / kg once every three weeks (q3w), or 6.4 mg / kg once every three weeks (q3w), but more preferably 4.8 mg / kg once every three weeks (q3w), 4.9 mg / kg once every three weeks (q3w), 5.0 mg / kg once every three weeks (q3w), 5.1 mg / kg once every three weeks (q3w), 5.2 mg / kg once every three weeks (q3w), 5.3 mg / kg once every three weeks (q3w), 5.4 mg / kg once every three weeks (q3w), 5.5 mg / kg once every three weeks (q3w), 5.It can be 6 mg / kg once every three weeks (q3w), 5.7 mg / kg once every three weeks (q3w), 5.8 mg / kg once every three weeks (q3w), 5.9 mg / kg once every three weeks (q3w), 6.0 mg / kg once every three weeks (q3w), 6.1 mg / kg once every three weeks (q3w), 6.2 mg / kg once every three weeks (q3w), 6.3 mg / kg once every three weeks (q3w), or 6.4 mg / kg once every three weeks (q3w).

[0359] In some embodiments, the dosing regimen is 5 mg once every three weeks (q3w), 10 mg once every three weeks (q3w), 15 mg once every three weeks (q3w), 20 mg once every three weeks (q3w), 25 mg once every three weeks (q3w), 30 mg once every three weeks (q3w), 35 mg once every three weeks (q3w), 40 mg once every three weeks (q3w), 45 mg once every three weeks (q3w), 50 mg once every three weeks (q3w), 55 mg once every three weeks (q3w), 60 mg once every three weeks (q3w), 65 mg once every three weeks (q3w), 70 mg once every three weeks (q3w), 75 mg once every three weeks (q3w), 80 mg once every three weeks (q3w), 85 mg once every three weeks (q3w), 90 mg once every three weeks (q3w), 95 mg once every three weeks (q3w), 100 mg once every three weeks (q3w), 125 mg once every three weeks (q3w), 150 mg once every three weeks (q3w), 175 mg once every three weeks (q3w), 200 mg once every three weeks (q3w), 205 mg once every three weeks (q3w), 210 mg once every three weeks (q3w), 215 mg once every three weeks (q3w), 220 mg once every three weeks (q3w), 225 mg once every three weeks (q3w), 230 mg once every three weeks (q3w), 235 mg once every three weeks (q3w), 240 mg once every three weeks (q3w), 245 mg once every three weeks (q3w), 250 mg once every three weeks (q3w), 255 mg once every three weeks (q3w), 260 mg once every three weeks (q3w), 265 mg once every three weeks (q3w), 270 mg once every three weeks (q3w), 275 mg once every three weeks (q3w), 280 mg once every three weeks (q3w), 285 mg once every three weeks (q3w), 290 mg once every three weeks (q3w), 295 mg once every three weeks (q3w), 300 mg once every three weeks (q3w), 305 mg once every three weeks (q3w), 310 mg once every three weeks (q3w), 315 mg once every three weeks (q3w), 320 mg once every three weeks (q3w), 325 mg once every three weeks (q3w), 330 mg once every three weeks (q3w),335 mg once every three weeks (q3w), 340 mg once every three weeks (q3w), 345 mg once every three weeks (q3w), 350 mg once every three weeks (q3w), 355 mg once every three weeks (q3w), 360 mg once every three weeks (q3w), 365 mg once every three weeks (q3w), 370 mg once every three weeks (q3w), 375 mg once every three weeks (q3w), 380 mg once every three weeks (q3w), 385 mg once every three weeks (q3w), 390 mg once every three weeks (q3w), 395 mg once every three weeks (q3w), 400 mg once every three weeks (q3w), 405 mg once every three weeks (q3w), 410 mg once every three weeks (q3w), 415 mg once every three weeks (q3w), 420 mg once every three weeks (q3w), 425 mg once every three weeks (q3w), 430 mg once every three weeks (q3w), 435 mg once every three weeks (q3w), 440 mg once every three weeks (q3w), 445 mg once every three weeks (q3w), 450 mg once every three weeks (q3w), 455 mg once every three weeks (q3w), 460 mg once every three weeks (q3w), 465 mg once every three weeks (q3w), 470 mg once every three weeks (q3w), 475 mg once every three weeks (q3w), 480 mg once every three weeks (q3w), 485 mg once every three weeks (q3w), 490 mg once every three weeks (q3w), 495 mg once every three weeks (q3w), 500 mg once every three weeks (q3w), 505 mg once every three weeks (q3w), 510 mg once every three weeks (q3w), 515 mg once every three weeks (q3w), 520 mg once every three weeks (q3w), 525 mg once every three weeks (q3w), 530 mg once every three weeks (q3w), 535 mg once every three weeks (q3w), 540 mg once every three weeks (q3w), 545 mg once every three weeks (q3w), 550 mg once every three weeks (q3w), 555 mg once every three weeks (q3w), 560 mg once every three weeks (q3w), 565 mg once every three weeks (q3w), 570 mg once every three weeks (q3w), 575 mg once every three weeks (q3w), 580 mg once every three weeks (q3w),585 mg once every three weeks (q3w), 590 mg once every three weeks (q3w), 595 mg once every three weeks (q3w), 600 mg once every three weeks (q3w), 605 mg once every three weeks (q3w), 610 mg once every three weeks (q3w), 615 mg once every three weeks (q3w), 620 mg once every three weeks (q3w), 625 mg once every three weeks (q3w), 630 mg once every three weeks (q3w), 635 mg once every three weeks (q3w), 640 mg once every three weeks (q3w), 645 mg once every three weeks (q3w), 650 mg once every three weeks (q3w), 655 mg once every three weeks (q3w), 660 mg once every three weeks (q3w), 665 mg once every three weeks (q3w), 670 mg once every three weeks (q3w), 675 mg once every three weeks (q3w), 680 mg once every three weeks (q3w), 685 mg once every three weeks (q3w), 690 mg once every three weeks (q3w), 695 mg once every three weeks (q3w), 700 mg once every three weeks (q3w), 705 mg once every three weeks (q3w), 710 mg once every three weeks (q3w), 715 mg once every three weeks (q3w), 720 mg once every three weeks (q3w), 725 mg once every three weeks (q3w), 730 mg once every three weeks (q3w), 735 mg once every three weeks (q3w), 740 mg once every three weeks (q3w), 745 mg once every three weeks (q3w), 750 mg once every three weeks (q3w), 755 mg once every three weeks (q3w), 760 mg once every three weeks (q3w), 765 mg once every three weeks (q3w), 770 mg once every three weeks (q3w), 775 mg once every three weeks (q3w), 780 mg once every three weeks (q3w), 785 mg once every three weeks (q3w), 790 mg once every three weeks (q3w), 795 mg once every three weeks (q3w), 800 mg once every three weeks (q3w), 805 mg once every three weeks (q3w), 810 mg once every three weeks (q3w), 815 mg once every three weeks (q3w), 820 mg once every three weeks (q3w), 825 mg once every three weeks (q3w), 830 mg once every three weeks (q3w),835 mg once every three weeks (q3w), 840 mg once every three weeks (q3w), 845 mg once every three weeks (q3w), 850 mg once every three weeks (q3w), 855 mg once every three weeks (q3w), 860 mg once every three weeks (q3w), 865 mg once every three weeks (q3w), 870 mg once every three weeks (q3w), 875 mg once every three weeks (q3w), 880 mg once every three weeks (q3w), 885 mg once every three weeks (q3w), 890 mg once every three weeks (q3w), 895 mg once every three weeks (q3w), 900 mg once every three weeks (q3w), 905 mg once every three weeks (q3w), 910 mg once every three weeks (q3w), 915 mg once every three weeks (q3w), 920 mg once every three weeks (q3w), 925 mg once every three weeks (q3w), 930 mg once every three weeks (q3w), 935 mg once every three weeks (q3w), 940 mg once every three weeks (q3w), 945 mg once every three weeks (q3w), 950 mg once every three weeks (q3w), 955 mg once every three weeks (q3w), 960 mg once every three weeks (q3w), 965 mg once every three weeks (q3w), 970 mg once every three weeks (q3w), 975 mg once every three weeks (q3w), 980 mg once every three weeks (q3w), 985 mg once every three weeks (q3w), 990 mg once every three weeks (q3w), 995 mg once every three weeks (q3w), 1000 mg once every three weeks (q3w), 1025 mg once every three weeks (q3w), 1050 mg once every three weeks (q3w), 1075 mg once every three weeks (q3w), 1100 mg once every three weeks (q3w), 1125 mg once every three weeks (q3w), 1150 mg once every three weeks (q3w), 1175 mg once every three weeks (q3w), 1200 mg once every three weeks (q3w), 1225 mg once every three weeks (q3w), 1250 mg once every three weeks (q3w), 1275 mg once every three weeks (q3w), 1300 mg once every three weeks (q3w), 1325 mg once every three weeks (q3w), 1350 mg once every three weeks (q3w), 1375 mg once every three weeks (q3w)1400 mg once every three weeks (q3w), 1425 mg once every three weeks (q3w), 1450 mg once every three weeks (q3w), 1475 mg once every three weeks (q3w), 1500 mg once every three weeks (q3w), 1525 mg once every three weeks (q3w), 1550 mg once every three weeks (q3w), 1575 mg once every three weeks (q3w), 1600 mg once every three weeks (q3w), 1625 mg once every three weeks (q3w), 1650 mg once every three weeks (q3w), 1675 mg once every three weeks (q3w), 1700 mg once every three weeks (q3w), 1725 mg once every three weeks (q3w), 1750 mg once every three weeks (q3w), 1775 mg once every three weeks (q3w), 1800 mg once every three weeks (q3w), 1825 mg once every three weeks (q3w), 1850 mg once every three weeks (q3w), 1875 mg once every three weeks (q3w), 1900 mg once every three weeks (q3w), 1925 mg once every three weeks (q3w), 1950 mg once every three weeks (q3w), 1975 mg once every three weeks (q3w), or 2000 mg once every three weeks (q3w) may be used.,

[0360] In a preferred embodiment, the dosing regimen is 200 mg once every three weeks (q3w), 205 mg once every three weeks (q3w), 210 mg once every three weeks (q3w), 215 mg once every three weeks (q3w), 220 mg once every three weeks (q3w), 225 mg once every three weeks (q3w), 230 mg once every three weeks (q3w), 235 mg once every three weeks (q3w), 240 mg once every three weeks (q3w), 245 mg once every three weeks (q3w), 250 mg once every three weeks (q3w), 255 mg once every three weeks (q3w), 260 mg once every three weeks (q3w), 265 mg once every three weeks (q3w), 270 mg once every three weeks (q3w), 275 mg once every three weeks (q3w), 280 mg once every three weeks (q3w), 285 mg once every three weeks (q3w), 290 mg once every three weeks (q3w), 295 mg once every three weeks (q3w), 300 mg once every three weeks (q3w), 305 mg once every three weeks (q3w), 310 mg once every three weeks (q3w), 315 mg once every three weeks (q3w), 320 mg once every three weeks (q3w), 325 mg once every three weeks (q3w), 330 mg once every three weeks (q3w), 335 mg once every three weeks (q3w), 340 mg once every three weeks (q3w), 345 mg once every three weeks (q3w), 350 mg once every three weeks (q3w), 355 mg once every three weeks (q3w), 360 mg once every three weeks (q3w), 365 mg once every three weeks (q3w), 370 mg once every three weeks (q3w), 375 mg once every three weeks (q3w), 380 mg once every three weeks (q3w), 385 mg once every three weeks (q3w), 390 mg once every three weeks (q3w), 395 mg once every three weeks (q3w), 400 mg once every three weeks (q3w), 405 mg once every three weeks (q3w), 410 mg once every three weeks (q3w), 415 mg once every three weeks (q3w), 420 mg once every three weeks (q3w), 425 mg once every three weeks (q3w), 430 mg once every three weeks (q3w), 435 mg once every three weeks (q3w), 440 mg once every three weeks (q3w),445 mg once every three weeks (q3w), 450 mg once every three weeks (q3w), 455 mg once every three weeks (q3w), 460 mg once every three weeks (q3w), 465 mg once every three weeks (q3w), 470 mg once every three weeks (q3w), 475 mg once every three weeks (q3w), 480 mg once every three weeks (q3w), 485 mg once every three weeks (q3w), 490 mg once every three weeks (q3w), 495 mg once every three weeks (q3w), 500 mg once every three weeks (q3w), 505 mg once every three weeks (q3w), 510 mg once every three weeks (q3w), 515 mg once every three weeks (q3w), 520 mg once every three weeks (q3w), 525 mg once every three weeks (q3w), 530 mg once every three weeks (q3w), 535 mg once every three weeks (q3w), 540 mg once every three weeks (q3w), 545 mg once every three weeks (q3w), 550 mg once every three weeks (q3w), 555 mg once every three weeks (q3w), 560 mg once every three weeks (q3w), 565 mg once every three weeks (q3w), 570 mg once every three weeks (q3w), 575 mg once every three weeks (q3w), 580 mg once every three weeks (q3w), 585 mg once every three weeks (q3w), 590 mg once every three weeks (q3w), 595 mg once every three weeks (q3w), 600 mg once every three weeks (q3w), 605 mg once every three weeks (q3w), 610 mg once every three weeks (q3w), 615 mg once every three weeks (q3w), 620 mg once every three weeks (q3w), 625 mg once every three weeks (q3w), 630 mg once every three weeks (q3w), 635 mg once every three weeks (q3w), 640 mg once every three weeks (q3w), 645 mg once every three weeks (q3w), 650 mg once every three weeks (q3w), 655 mg once every three weeks (q3w), 660 mg once every three weeks (q3w), 665 mg once every three weeks (q3w), 670 mg once every three weeks (q3w), 675 mg once every three weeks (q3w), 680 mg once every three weeks (q3w), 685 mg once every three weeks (q3w), 690 mg once every three weeks (q3w),695 mg once every three weeks (q3w), 700 mg once every three weeks (q3w), 705 mg once every three weeks (q3w), 710 mg once every three weeks (q3w), 715 mg once every three weeks (q3w), 720 mg once every three weeks (q3w), 725 mg once every three weeks (q3w), 730 mg once every three weeks (q3w), 735 mg once every three weeks (q3w), 740 mg once every three weeks (q3w), 745 mg once every three weeks (q3w), 750 mg once every three weeks (q3w), 755 mg once every three weeks (q3w), 760 mg once every three weeks (q3w), 765 mg once every three weeks (q3w), 770 mg once every three weeks (q3w), 775 mg once every three weeks (q3w), 780 mg once every three weeks (q3w), 785 mg once every three weeks (q3w), 790 mg once every three weeks (q3w), 795 mg once every three weeks (q3w), 800 mg once every three weeks (q3w), 805 mg once every three weeks (q3w), 810 mg once every three weeks (q3w), 815 mg once every three weeks (q3w), 820 mg once every three weeks (q3w), 825 mg once every three weeks (q3w), 830 mg once every three weeks (q3w), 835 mg once every three weeks (q3w), 840 mg once every three weeks (q3w), 845 mg once every three weeks (q3w), 850 mg once every three weeks (q3w), 855 mg once every three weeks (q3w), 860 mg once every three weeks (q3w), 865 mg once every three weeks (q3w), 870 mg once every three weeks (q3w), 875 mg once every three weeks (q3w), 880 mg once every three weeks (q3w), 885 mg once every three weeks (q3w), 890 mg once every three weeks (q3w), 895 mg once every three weeks (q3w), 900 mg once every three weeks (q3w), 905 mg once every three weeks (q3w), 910 mg once every three weeks (q3w), 915 mg once every three weeks (q3w), 920 mg once every three weeks (q3w), 925 mg once every three weeks (q3w), 930 mg once every three weeks (q3w), 935 mg once every three weeks (q3w), 940 mg once every three weeks (q3w)It can be 945 mg once every three weeks (q3w), 950 mg once every three weeks (q3w), 955 mg once every three weeks (q3w), 960 mg once every three weeks (q3w), 965 mg once every three weeks (q3w), 970 mg once every three weeks (q3w), 975 mg once every three weeks (q3w), 980 mg once every three weeks (q3w), 985 mg once every three weeks (q3w), 990 mg once every three weeks (q3w), 995 mg once every three weeks (q3w), 1000 mg once every three weeks (q3w). Also, in some embodiments, a single bolus can be administered, while in some embodiments, multiple divided doses can be administered over time, or the dose can be proportionally decreased or increased as indicated by the circumstances.,

[0361] Furthermore, the subjects of the methods and uses are mostly cancer patients, but the age of the patients is not limited. The disclosed methods and uses are useful for treating cancer, malignant diseases, or cancer cell proliferation having various recurrence and prognostic outcomes across all age groups and cohorts. Thus, in some embodiments, the subject can be a pediatric subject, while in other embodiments, the subject can be an adult subject.,

[0362] In some embodiments, the objective response rate of a subject administered the ADC of the present disclosure is at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, or about 80%. In a preferred embodiment, the objective response rate of a subject administered the ADC of the present disclosure is at least about 20%. In a more preferred embodiment, the objective response rate of a subject administered the ADC of the present disclosure is at least about 30%. In a more preferred embodiment, the objective response rate of a subject administered the ADC of the present disclosure is at least about 40%. In a more preferred embodiment, the objective response ...

Claims

1. A pharmaceutical composition for use in treating or preventing cancer, comprising an anti-CDH6 antibody-drug conjugate, The antibody-drug conjugate comprises an anti-CDH6 antibody and an anti-tumor compound connected by a linker; The linker and the antitumor compound have the following formula: -(Succinimid-3-yl-N)-CH ２ CH ２ CH ２ CH ２ CH ２ -C(=O)-GGFG-NH-CH ２ -O-CH ２ -C(=O)-(NH-DX) is represented by In the formula, -(Succinimid-3-yl-N)- is a group represented by the following formula: 【Chemistry 1】 and (NH-DX) has a structure represented by the following formula: 【Chemistry 2】 wherein the nitrogen atom of the amino group at position 1 is the attachment position; The anti-CDH6 antibody comprises, in its heavy chain variable region, a CDRH1 consisting of the amino acid sequence of SEQ ID NO: 17, a CDRH2 consisting of the amino acid sequence of SEQ ID NO: 60, and a CDRH3 consisting of the amino acid sequence of SEQ ID NO: 19, and, in its light chain variable region, a CDRL1 consisting of the amino acid sequence of SEQ ID NO: 12, a CDRL2 consisting of the amino acid sequence of SEQ ID NO: 13, and a CDRL3 consisting of the amino acid sequence of SEQ ID NO: 14; The pharmaceutical composition wherein a dose of the antibody-drug conjugate in the range of 3.2 mg / kg to 9.6 mg / kg is administered to the subject.

2. The pharmaceutical composition described in claim 1, wherein the anti-CDH6 antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 71 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

63.

3. The pharmaceutical composition of claim 1, wherein the anti-CDH6 antibody comprises a heavy chain comprising the amino acid sequence of positions 20 to 471 in SEQ ID NO: 69 and a light chain comprising the amino acid sequence of positions 21 to 233 in SEQ ID NO:

61.

4. The pharmaceutical composition described in claim 3, wherein the anti-CDH6 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain.

5. The antibody-drug conjugate has the following formula: 【Transformation 3】 and having a structure represented by wherein Ab is an antibody comprising a heavy chain variable region comprising SEQ ID NO: 71 and a light chain variable region comprising SEQ ID NO: 63; n represents the average number of drug-linker structures bound to an antibody per antibody, and is within the range of 7 to 8; The pharmaceutical composition of claim 1.

6. The pharmaceutical composition of claim 5, wherein the antibody comprises a heavy chain comprising the amino acid sequence of positions 20 to 471 in SEQ ID NO: 69 and a light chain comprising the amino acid sequence of positions 21 to 233 in SEQ ID NO:

61.

7. The pharmaceutical composition of claim 6, wherein the anti-CDH6 antibody lacks a lysine residue at the carboxyl terminus of the heavy chain.

8. 10. The pharmaceutical composition of claim 1, wherein a dose of the antibody-drug conjugate selected from the group consisting of 4.8 mg / kg to 9.6 mg / kg, 3.2 mg / kg to 8.0 mg / kg, 3.2 mg / kg to 6.4 mg / kg, 4.8 mg / kg to 8.0 mg / kg, and 4.8 mg / kg to 6.4 mg / kg is administered to the subject.

9. 10. The pharmaceutical composition of claim 1, wherein a dose of the antibody-drug conjugate selected from the group consisting of about 3.2 mg / kg, about 4.8 mg / kg, about 5.4 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg, and about 8.0 mg / kg is administered to the subject.

10. 3.2mg / kg, 3.3mg / kg, 3.4mg / kg, 3.5mg / kg, 3.6mg / kg, 3.7mg / kg, 3.8mg / kg, 3.9mg / kg, 4.0mg / kg, 4.1mg / kg, 4.2mg / kg, 4.3mg / kg, 4.4mg / kg, 4.5mg / kg, 4.6mg / kg, 4.7mg / kg, 4.8mg / kg, 4.9mg / kg, 5.0mg / kg, 5.1m 10. The pharmaceutical composition of claim 1, wherein a dose of the antibody-drug conjugate selected from the group consisting of: 5.2 mg / kg, 5.3 mg / kg, 5.4 mg / kg, 5.5 mg / kg, 5.6 mg / kg, 5.7 mg / kg, 5.8 mg / kg, 5.9 mg / kg, 6.0 mg / kg, 6.1 mg / kg, 6.2 mg / kg, 6.3 mg / kg, and 6.4 mg / kg is administered to the subject.

11. The pharmaceutical composition of claim 1, wherein a dose of the antibody-drug conjugate selected from the group consisting of 4.8 mg / kg, 4.9 mg / kg, 5.0 mg / kg, 5.1 mg / kg, 5.2 mg / kg, 5.3 mg / kg, 5.4 mg / kg, 5.5 mg / kg, 5.6 mg / kg, 5.7 mg / kg, 5.8 mg / kg, 5.9 mg / kg, 6.0 mg / kg, 6.1 mg / kg, 6.2 mg / kg, 6.3 mg / kg, and 6.4 mg / kg is administered to the subject.

12. The pharmaceutical composition of claim 5, wherein a dose of 4.8 mg / kg of the antibody-drug conjugate is administered to the subject.

13. The pharmaceutical composition of claim 6, wherein a dose of 4.8 mg / kg of the antibody-drug conjugate is administered to the subject.

14. The pharmaceutical composition of claim 7, wherein a dose of 4.8 mg / kg of the antibody-drug conjugate is administered to the subject.

15. The pharmaceutical composition of claim 5, wherein a dose of 5.6 mg / kg of the antibody-drug conjugate is administered to the subject.

16. The pharmaceutical composition of claim 6, wherein a dose of 5.6 mg / kg of the antibody-drug conjugate is administered to the subject.

17. The pharmaceutical composition of claim 7, wherein a dose of 5.6 mg / kg of the antibody-drug conjugate is administered to the subject.

18. The pharmaceutical composition of claim 5, wherein a dose of 6.4 mg / kg of the antibody-drug conjugate is administered to the subject.

19. The pharmaceutical composition of claim 6, wherein a dose of 6.4 mg / kg of the antibody-drug conjugate is administered to the subject.

20. The pharmaceutical composition of claim 7, wherein a dose of 6.4 mg / kg of the antibody-drug conjugate is administered to the subject.

21. The pharmaceutical composition of claim 1, wherein the antibody-drug conjugate is administered intravenously.

22. The pharmaceutical composition of claim 12, wherein the antibody-drug conjugate is administered intravenously.

23. The pharmaceutical composition of claim 13, wherein the antibody-drug conjugate is administered intravenously.

24. The pharmaceutical composition of claim 14, wherein the antibody-drug conjugate is administered intravenously.

25. The pharmaceutical composition of claim 15, wherein the antibody-drug conjugate is administered intravenously.

26. The pharmaceutical composition of claim 16, wherein the antibody-drug conjugate is administered intravenously.

27. The pharmaceutical composition of claim 17, wherein the antibody-drug conjugate is administered intravenously.

28. The pharmaceutical composition of claim 18, wherein the antibody-drug conjugate is administered intravenously.

29. The pharmaceutical composition of claim 19, wherein the antibody-drug conjugate is administered intravenously.

30. The pharmaceutical composition of claim 20, wherein the antibody-drug conjugate is administered intravenously.

31. The pharmaceutical composition of claim 1, wherein the antibody-drug conjugate is administered once every three weeks.

32. The pharmaceutical composition of claim 22, wherein the antibody-drug conjugate is administered once every three weeks.

33. The pharmaceutical composition of claim 23, wherein the antibody-drug conjugate is administered once every three weeks.

34. The pharmaceutical composition of claim 24, wherein the antibody-drug conjugate is administered once every three weeks.

35. The pharmaceutical composition of claim 25, wherein the antibody-drug conjugate is administered once every three weeks.

36. The pharmaceutical composition of claim 26, wherein the antibody-drug conjugate is administered once every three weeks.

37. The pharmaceutical composition of claim 27, wherein the antibody-drug conjugate is administered once every three weeks.

38. The pharmaceutical composition of claim 28, wherein the antibody-drug conjugate is administered once every three weeks.

39. The pharmaceutical composition of claim 29, wherein the antibody-drug conjugate is administered once every three weeks.

40. The pharmaceutical composition of claim 30, wherein the antibody-drug conjugate is administered once every three weeks.

41. 2. The pharmaceutical composition of claim 1, wherein the cancer is selected from the group consisting of renal cell carcinoma, renal clear cell carcinoma, papillary renal cell carcinoma, ovarian cancer, ovarian serous adenocarcinoma, ovarian clear cell carcinoma, endometrioid ovarian carcinoma, ovarian mucinous tumor, thyroid cancer, bile duct cancer, lung cancer, non-small cell lung cancer, cervical cancer, brain tumor, head and neck cancer, sarcoma, osteosarcoma, small cell lung cancer, glioblastoma, mesothelioma, uterine cancer, pancreatic cancer, Wilms' tumor, neuroblastoma, colorectal cancer, gastric cancer, endometrial cancer, nasopharyngeal cancer, prostate cancer, and cancers associated with von Hippel-Lindau disease.

42. 33. The pharmaceutical composition of claim 32, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

43. The pharmaceutical composition described in claim 33, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

44. The pharmaceutical composition described in claim 34, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

45. The pharmaceutical composition described in claim 35, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

46. The pharmaceutical composition described in claim 36, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

47. The pharmaceutical composition described in claim 37, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

48. The pharmaceutical composition described in claim 38, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

49. The pharmaceutical composition described in claim 39, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

50. The pharmaceutical composition described in claim 40, wherein the cancer is renal cell carcinoma, renal clear cell carcinoma, or papillary renal cell carcinoma.

51. 33. The pharmaceutical composition of claim 32, wherein the cancer is ovarian cancer.

52. The pharmaceutical composition described in claim 33, wherein the cancer is ovarian cancer.

53. The pharmaceutical composition described in claim 34, wherein the cancer is ovarian cancer.

54. The pharmaceutical composition described in claim 35, wherein the cancer is ovarian cancer.

55. The pharmaceutical composition described in claim 36, wherein the cancer is ovarian cancer.

56. The pharmaceutical composition described in claim 37, wherein the cancer is ovarian cancer.

57. The pharmaceutical composition described in claim 38, wherein the cancer is ovarian cancer.

58. The pharmaceutical composition described in claim 39, wherein the cancer is ovarian cancer.

59. The pharmaceutical composition described in claim 40, wherein the cancer is ovarian cancer.

60. 52. The pharmaceutical composition of claim 51, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

61. The pharmaceutical composition described in claim 52, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

62. The pharmaceutical composition described in claim 53, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

63. The pharmaceutical composition described in claim 54, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

64. The pharmaceutical composition described in claim 55, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

65. The pharmaceutical composition described in claim 56, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

66. The pharmaceutical composition described in claim 57, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

67. The pharmaceutical composition described in claim 58, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

68. The pharmaceutical composition described in claim 59, wherein the ovarian cancer is selected from the group consisting of epithelial ovarian cancer, fallopian tube cancer, and primary peritoneal cancer.

69. 69. The pharmaceutical composition of any one of claims 1 to 68, wherein the cancer is metastatic.

70. 69. The pharmaceutical composition of any one of claims 1 to 68, wherein the cancer is resistant or refractory.

71. The pharmaceutical composition of claim 70, wherein the resistance or refractory state is resistance or refractory state acquired by the cancer due to treatment with an anticancer drug.

72. 72. The pharmaceutical composition of claim 71, wherein the anticancer drug is a platinum-based chemotherapy drug, a chemotherapy drug, a PARP inhibitor, an immune checkpoint inhibitor, an angiogenesis inhibitor, or a VEGFR-TKI.

73. The pharmaceutical composition of any one of claims 1 to 68, wherein the subject has a history of treatment with one or more anticancer drugs selected from the group consisting of platinum-based chemotherapy drugs, chemotherapy drugs, PARP inhibitors, immune checkpoint inhibitors, angiogenesis inhibitors, and VEGFR-TKIs.

74. 69. The pharmaceutical composition of any one of claims 1 to 68, wherein the cancer is a CDH6-expressing cancer or a CDH6-overexpressing cancer.

75. 69. The pharmaceutical composition of any one of claims 1 to 68, wherein the cancer is inoperable or recurrent cancer.

76. the subject exhibited a recurrence of cancer prior to administration of the antibody-drug conjugate; The recurrence of cancer is (1) Occurring less than or within about 6 months of the completion of a chemotherapy regimen of (a) or (b) below: (a) a chemotherapy regimen including a platinum-based drug; (b) chemotherapy regimens including platinum-based drugs and taxanes; or (2) Occurring at or about 6 months after completion of a chemotherapy regimen of (a) or (b) below: (a) a chemotherapy regimen including a platinum-based drug; (b) chemotherapy regimens including platinum-based drugs and taxanes; 69. A pharmaceutical composition according to any one of claims 1 to 68.

77. 69. The pharmaceutical composition of any one of claims 1 to 68, administered as monotherapy.

78. 69. The pharmaceutical composition of any one of claims 1 to 68, wherein the subject is administered the pharmaceutical composition together with a second drug, wherein the pharmaceutical composition is administered before, after, or simultaneously with the second drug.

79. 69. The pharmaceutical composition of any one of claims 1 to 68, administered as a therapy selected from the group consisting of: (a) maintenance therapy, (b) adjuvant therapy, (c) after surgical resection of a tumor, (d) neoadjuvant therapy, and (e) before surgical resection of a tumor.

80. The following formula: 【Chemistry 4】 A pharmaceutical composition for use in treating or preventing cancer, comprising an anti-CDH6 antibody-drug conjugate having a structure represented by: In the formula, AB represents an anti-CDH6 antibody; n represents the average number of drug-linker structures bound to an antibody per antibody, The antibody is connected to the linker via a sulfhydryl group derived from the antibody, n is 7 to 8; The anti-CDH6 antibody comprises a heavy chain amino acid sequence represented by SEQ ID NO: 87, or an amino acid sequence derived from the amino acid sequence represented by SEQ ID NO: 87, with one or two amino acids deleted from the carboxyl terminus thereof; and a light chain amino acid sequence represented by SEQ ID NO: 88, the dose of the antibody-drug conjugate is in the range of 3.2 mg / kg to 9.6 mg / kg; The antibody-drug conjugate is administered intravenously once every three weeks; Optionally, the anti-CDH6 antibody-drug conjugate is a salt thereof, or a hydrate of the anti-CDH6 antibody-drug conjugate or a salt thereof. Pharmaceutical compositions.

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