Combination of Anti-EGFR antibody-drug conjugate with other agent

Abstract

[Problem] To provide a novel drug combination therapy for diseases, in particular, cancer. [Solution] Provided is a combination administration of an anti-EGFR antibody-drug conjugate with another agent such as an immune checkpoint inhibitor.

Description

Combination of anti-EGFR antibody-drug conjugates with other agents

[0001] The present invention relates to pharmaceutical compositions characterized by the use of a specific anti-EGFR antibody-drug conjugate in combination with another drug such as an immune checkpoint inhibitor, and therapeutic methods characterized by administering a specific anti-EGFR antibody-drug conjugate in combination with another drug such as an immune checkpoint inhibitor to a subject, etc.

[0002] In cancer treatment, removal of the cancerous tissue through surgery or killing of cancer cells with anticancer drugs or radiation is expected to result in a cure or prolong life. However, for many cancers, a single anticancer drug treatment alone is insufficient, and therefore treatments using combinations with various drugs have been attempted. Anticancer drugs include chemotherapeutic agents such as paclitaxel, capecitabine, irinotecan, doxorubicin, carboplatin, taxol, or camptothecin (Non-Patent Document 1), molecularly targeted drugs such as imatinib, crizotinib, dasatinib, and lapatinib, cancer therapeutic antibodies such as trastuzumab (Patent Document 1), bevacizumab, cetuximab, and ramucirumab, and antibody-drug conjugates such as trastuzumab emtansine (Patent Document 2). The anticancer drug and combination therapy used are determined depending on the type and stage of cancer and the treatment status.

[0003] Immune checkpoint inhibitors, which have recently become one of the standard treatments for some cancers, are drugs that inhibit the immunosuppressive system and activate antitumor immunity (Non-Patent Documents 2 to 4). Examples of immune checkpoint inhibitors include anti-PD-1 antibodies such as nivolumab (Patent Document 3), pembrolizumab (Patent Document 4), spartalizumab (Patent Document 5), cemiplimab (Patent Document 6), and dostarlimab, as well as anti-PD-L1 antibodies such as atezolizumab (Patent Document 7) and durvalumab. b) (Patent Document 8), Avelumab (Patent Document 9), anti-CTLA-4 antibodies Ipilimumab (Patent Document 10) and Tremelimumab (Patent Document 11), anti-TIGIT antibodies Tiragolumab (Patent Document 12) and Vibostolimab (Patent Document 13), and anti-LAG-3 antibody Relatlimab (Patent Document 14), etc. are known.

[0004] EGFR is known as an important target in cancer therapy (Non-Patent Document 5). Furthermore, EGFR mutations, mutations in EGFR downstream signaling molecules, or EGFR overexpression have been observed in several cancer types, and these have been shown to correlate with treatment resistance and poor prognosis (Non-Patent Documents 6-8). STING (Stimulator of Interferon Genes) is a transmembrane adaptor protein localized in the endoplasmic reticulum (Non-Patent Document 9). STING functions as a central molecule in innate immune activation in mammals and serves as the first line of defense against the invasion of pathogens such as bacteria and viruses. STING activation is known to be triggered by signals generated by multiple cytoplasmic DNA sensors upon sensing exogenous and endogenous DNA (Non-Patent Documents 10-12). Recent studies have shown that STING not only promotes host defense against microorganisms but also promotes antitumor immunity (Non-Patent Document 13). Patent Documents 15 to 18 describe conjugates in which a STING agonist is used as an immunostimulatory compound and is linked to an antibody via a linker, and examples of in vivo administration are described. Patent Document 19 describes examples in which a conjugate in which a STING agonist and an antibody are linked via a linker is combined with trastuzumab or an anti-PD-1 antibody and administered in vivo. Furthermore, Non-Patent Document 14 describes a test in which a conjugate in which a STING agonist and an anti-EGFR antibody are linked via a linker is combined with an anti-PD-L1 antibody and administered in vivo.

[0005] However, the effect of combining a conjugate of a STING agonist having a specific cyclic dinucleotide structure with an antibody (Patent Documents 17 and 18) with other drugs is unknown.

[0006] U.S. Patent No. 5,821,337, International Publication No. WO 2001 / 000244, International Publication No. WO 2006 / 121168, International Publication No. WO 2008 / 156712, International Publication No. WO 2015 / 112900, International Publication No. WO 2015 / 196051, International Publication No. WO 2010 / 077634, International Publication No. WO 2011 / 066389, International Publication No. WO 2013 / 079174, International Publication No. WO 2001 / 014424 International Publication No. 2000 / 037504 International Publication No. 2017 / 053748 International Publication No. 2016 / 028656 International Publication No. 2014 / 008218 International Publication No. 2021 / 202984 International Publication No. 2022 / 097117 International Publication No. 2020 / 050406 International Publication No. 2021 / 177438 International Publication No. 2023 / 172906

[0007] Genes & Diseases 2023, 10, 1367-1401. Cancers2016, 8, 106. Nat. Rev. Cancer 2012, 12, 252-264. Cell 2015, 162, 937. Front. Oncol. 2019,9,800Clin. Cancer. Res. 2013, 19(8), 2240-2247. J. Biomed. Sci. 2012, 19(1), 40. Clin. Cancer. Res. 2015, 21(15), 3377-3383. Nature 2008, 455, 674-678. Mol. Cell 2013, 51, 226-235. Science 2015, 347, aaa2630. J. Virol. 2014, 88, 5328-5341. Immunity 2014, 41, 830-842. Proc. Natl. Acad. Sci. USA 2022, 119 (49), e2214278119.

[0008] An objective of the present invention is to provide a novel drug combination therapy for diseases, particularly cancer.

[0009] The present inventors have conducted extensive research to solve the above problems and have found that the combined administration of an anti-EGFR antibody-drug conjugate of the following formula (I) with another drug such as an immune checkpoint inhibitor safely exhibits excellent combined effects (particularly, significant antitumor effects), thereby completing the present invention. Specifically, the present invention provides the following [A1] to [D33].

[0010] [A1] a) A pharmaceutical composition comprising an anti-EGFR antibody-drug conjugate, wherein a) the anti-EGFR antibody-drug conjugate and b) one or more selected from an immune checkpoint inhibitor, a chemotherapeutic agent, and a molecular targeted drug are used in combination, and the anti-EGFR antibody-drug conjugate is represented by the following formula (I): (In the formula, m ２ represents an integer of 1 or 2, L represents a linker connecting the N297 sugar chain and D, -Lb-La-Lp-Lc-* (wherein the asterisk indicates binding to drug D, and Lb is: (In the structural formula of Lb shown above, the asterisk indicates bonding to La, and the wavy line indicates bonding to the N297 sugar chain), where La is -C(=O)-CH ２ CH ２ Lp represents -GGFG- or -GGPI-, Lc represents -NH-CH ２ -), Ab is an anti-EGFR antibody or an antigen-binding fragment thereof comprising a heavy chain comprising CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and a light chain comprising CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, CDRL2 consisting of the amino acid sequence represented by DAS, and CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 7, wherein the N297 sugar chain is N297-(Fuc)MSG1 having the structure represented by the following formula: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２-NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of the 1-3 chain side of the branched chain of β-Man in the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1- or 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), or N297-(Fuc)SG: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), and D represents (wherein the asterisk indicates binding to L, and W represents -NH-). [A1'] The pharmaceutical composition according to [A1], wherein Ab is an anti-EGFR antibody comprising an Fc region (wild-type or mutant-type) of the antibody, or an antigen-binding fragment of the antibody. [A2] The pharmaceutical composition according to [A1], wherein D is (wherein the asterisk indicates binding to L). [A3] The pharmaceutical composition according to [A1], wherein the anti-EGFR antibody-drug conjugate is represented by the following formula: (Wherein Ab, N297 sugar chain and m ２ [A4] The pharmaceutical composition according to [A1] or [A2], wherein the anti-EGFR antibody-drug conjugate is represented by the following formula: In the formula, m ２is 2, the Ab comprises a heavy chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and a light chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2, and the N297 sugar chain is N297-(Fuc)SG having a structure represented by the following formula: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, the asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring, and n ５[A5] The pharmaceutical composition according to any one of [A1] to [A3], wherein the Ab comprises a heavy chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 119 of SEQ ID NO: 1, and a light chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 107 of SEQ ID NO: 2. [A6] The pharmaceutical composition according to any one of [A1] to [A5], wherein the Ab comprises a heavy chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by SEQ ID NO: 1, and a light chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by SEQ ID NO: 2. [A7] The pharmaceutical composition according to any one of [A1] to [A6], wherein the Ab comprises a heavy chain consisting of the amino acid sequence represented by SEQ ID NO: 1 and a light chain consisting of the amino acid sequence represented by SEQ ID NO: 2. [A8] The pharmaceutical composition according to any one of [A1] to [A5], wherein the Ab comprises an antibody or an antigen-binding fragment of the antibody contained in the pharmaceutical composition according to any one of [A1] to [A7], comprising: (i) a heavy chain consisting of an amino acid sequence in which one or two amino acids are deleted from the carboxyl terminus of the amino acid sequence of the heavy chain; and (ii) a light chain. [A9] The pharmaceutical composition according to any one of [A1] to [A8], wherein a) an anti-EGFR antibody-drug conjugate and b) an immune checkpoint inhibitor are used in combination, and the immune checkpoint inhibitor is an antibody selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-TIGIT antibody, and an anti-LAG-3 antibody, an antigen-binding fragment thereof, or a compound comprising any of them. [A10] The pharmaceutical composition according to [A9], wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, cemiplimab, or dostarlimab. [A11] The pharmaceutical composition according to [A9], wherein the anti-PD-L1 antibody is atezolizumab, duvalumab, or avelumab. [A12] The pharmaceutical composition according to [A9], wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab. [A13] The pharmaceutical composition according to [A9], wherein the anti-TIGIT antibody is tiragolumab or vibostolimab.[A14] The pharmaceutical composition according to [A9], wherein the anti-LAG-3 antibody is leratolimab. [A15] The pharmaceutical composition according to any one of [A1] to [A8], wherein a) an anti-EGFR antibody-drug conjugate and b) a chemotherapeutic agent are used in combination, and the chemotherapeutic agent is an antimetabolite, a tubulin inhibitor, an alkylating agent, a platinum compound, or a topoisomerase inhibitor. [A16] The pharmaceutical composition according to [A15], wherein the antimetabolite is gemcitabine or a pharmaceutically acceptable salt thereof. [A17] The pharmaceutical composition according to [A15], wherein the antimetabolite is fluorouracil or a pharmaceutically acceptable salt thereof. [A18] The pharmaceutical composition according to [A15], wherein the tubulin inhibitor is docetaxel or a pharmaceutically acceptable salt thereof. [A19] The pharmaceutical composition according to [A15], wherein the platinum compound is oxaliplatin or a pharmaceutically acceptable salt thereof. [A20] The pharmaceutical composition according to [A15], wherein the platinum agent is cisplatin or a pharmaceutically acceptable salt thereof. [A21] The pharmaceutical composition according to [A15], wherein the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof. [A22] The pharmaceutical composition according to any one of [A1] to [A8], wherein a) an anti-EGFR antibody-drug conjugate and b) a molecularly targeted drug are used in combination, and the molecularly targeted drug is a VEGF inhibitor or an EGFR inhibitor. [A23] The pharmaceutical composition according to [A22], wherein the VEGF inhibitor is bevacizumab, ramucirumab, aflibercept beta, or axitinib. [A24] The pharmaceutical composition according to [A22], wherein the EGFR inhibitor is osimertinib or a pharmaceutically acceptable salt thereof. [A25] The pharmaceutical composition according to any one of [A1] to [A24], wherein a) the anti-EGFR antibody-drug conjugate; and b) the immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug are contained as active ingredients in separate formulations, and are administered simultaneously (simultaneously) or at different times (asynchronously or separately). [A26] The pharmaceutical composition according to any one of [A1] to [A25], for the treatment of cancer.[A27] The pharmaceutical composition according to [A26], wherein the cancer is at least one selected from the group consisting of lung cancer, colon cancer, kidney cancer, liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, cervical cancer, head and neck cancer, bile duct cancer, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule. [A28] The pharmaceutical composition according to [A27], wherein the cancer is lung cancer. [A29] The pharmaceutical composition according to [A27], wherein the cancer is colon cancer. [A30] The pharmaceutical composition according to [A27], wherein the cancer is breast cancer. [A31] The pharmaceutical composition according to [A27], wherein the cancer is head and neck cancer. [A32] The pharmaceutical composition according to [A27], wherein the cancer is kidney cancer. [A33] The pharmaceutical composition according to [A27], wherein the cancer is liver cancer. [B1] An anti-EGFR antibody-drug conjugate for use in combination with one or more selected from an immune checkpoint inhibitor, a chemotherapeutic agent, and a molecular targeted drug for the treatment of cancer, wherein the anti-EGFR antibody-drug conjugate is represented by the following formula (I): (In the formula, m ２ represents an integer of 1 or 2, L represents a linker connecting the N297 sugar chain and D, -Lb-La-Lp-Lc-* (wherein the asterisk indicates binding to drug D, and Lb is: (In the structural formula of Lb shown above, the asterisk indicates bonding to La, and the wavy line indicates bonding to the N297 sugar chain), where La is -C(=O)-CH ２ CH ２ Lp represents -GGFG- or -GGPI-, Lc represents -NH-CH ２-), the Ab is an anti-EGFR antibody or an antigen-binding fragment thereof comprising a heavy chain comprising a CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and a light chain comprising a CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, a CDRL2 consisting of the amino acid sequence represented by DAS, and a CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 7, wherein the N297 sugar chain is N297-(Fuc)MSG1 having the structure represented by the following formula: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of the 1-3 chain side of the branched chain of β-Man in the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1- or 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), or N297-(Fuc)SG: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), and D represents (wherein the asterisk indicates binding to L, and W represents -NH-). [B1'] The anti-EGFR antibody-drug conjugate according to [B1], wherein Ab is an anti-EGFR antibody comprising an Fc region (wild-type or mutant-type) of the antibody, or an antigen-binding fragment of the antibody. [B2] D is (wherein the asterisk indicates binding to L). [B3] The anti-EGFR antibody-drug conjugate according to [B1], represented by the following formula: (Wherein Ab, N297 sugar chain and m ２ [B4] The anti-EGFR antibody-drug conjugate according to [B1] or [B2], wherein the anti-EGFR antibody-drug conjugate is represented by the following formula: In the formula, m ２ is 2, the Ab comprises a heavy chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and a light chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2, and the N297 sugar chain is N297-(Fuc)SG having a structure represented by the following formula: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, the asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring, and n ５represents an integer of 3). [B5] The anti-EGFR antibody-drug conjugate of any one of [B1] to [B4], wherein the Ab comprises a heavy chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 119 of SEQ ID NO: 1, and a light chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 107 of SEQ ID NO: 2. [B6] The anti-EGFR antibody-drug conjugate of any one of [B1] to [B5], wherein the Ab comprises a heavy chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by SEQ ID NO: 1, and a light chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by SEQ ID NO: 2. [B7] The anti-EGFR antibody-drug conjugate according to any one of [B1] to [B6], wherein the Ab comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 1 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 2. [B8] The anti-EGFR antibody-drug conjugate according to any one of [B1] to [B7], wherein the Ab comprises an antibody or an antigen-binding fragment of said antibody comprised in the anti-EGFR antibody-drug conjugate according to any one of [B1] to [B7], comprising: (i) a heavy chain consisting of an amino acid sequence in which one or two amino acids are deleted from the carboxyl terminus of the amino acid sequence of the heavy chain; and (ii) a light chain. [B9] An anti-EGFR antibody-drug conjugate according to any one of [B1] to [B8] for use in combination with an immune checkpoint inhibitor for the treatment of cancer, wherein the immune checkpoint inhibitor is an antibody selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-TIGIT antibody, and an anti-LAG-3 antibody, an antigen-binding fragment thereof, or a compound comprising any of them. [B10] The anti-EGFR antibody-drug conjugate according to [B9], wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, cemiplimab, or dostarlimab.[B11] The anti-EGFR antibody-drug conjugate according to [B9], wherein the anti-PD-L1 antibody is atezolizumab, duvalumab, or avelumab. [B12] The anti-EGFR antibody-drug conjugate according to [B9], wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab. [B13] The anti-EGFR antibody-drug conjugate according to [B9], wherein the anti-TIGIT antibody is tiragolumab or vibostolimab. [B14] The anti-EGFR antibody-drug conjugate according to [B9], wherein the anti-LAG-3 antibody is leratolimab. [B15] An anti-EGFR antibody-drug conjugate according to any one of [B1] to [B8] for use in combination with a chemotherapeutic agent for the treatment of cancer, wherein the chemotherapeutic agent is an antimetabolite, a tubulin inhibitor, an alkylating agent, a platinum compound, or a topoisomerase inhibitor. [B16] The anti-EGFR antibody-drug conjugate according to [B15], wherein the antimetabolite is gemcitabine or a pharmaceutically acceptable salt thereof. [B17] The anti-EGFR antibody-drug conjugate according to [B15], wherein the antimetabolite is fluorouracil or a pharmaceutically acceptable salt thereof. [B18] The anti-EGFR antibody-drug conjugate according to [B15], wherein the tubulin inhibitor is docetaxel or a pharmaceutically acceptable salt thereof. [B19] The anti-EGFR antibody-drug conjugate according to [B15], wherein the platinum agent is oxaliplatin or a pharmaceutically acceptable salt thereof. [B20] The anti-EGFR antibody-drug conjugate according to [B15], wherein the platinum agent is cisplatin or a pharmaceutically acceptable salt thereof. [B21] The anti-EGFR antibody-drug conjugate according to [B15], wherein the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof. [B22] The anti-EGFR antibody-drug conjugate according to any one of [B1] to [B8], for use in combination with a molecular targeted drug for the treatment of cancer, wherein the molecular targeted drug is a VEGF inhibitor or an EGFR inhibitor.[B23] The anti-EGFR antibody-drug conjugate according to [B22], wherein the VEGF inhibitor is bevacizumab, ramucirumab, aflibercept beta, or axitinib. [B24] The anti-EGFR antibody-drug conjugate according to [B22], wherein the EGFR inhibitor is osimertinib or a pharmaceutically acceptable salt thereof. [B25] The anti-EGFR antibody-drug conjugate according to any one of [B1] to [B24], wherein the anti-EGFR antibody-drug conjugate; and the immune checkpoint inhibitor, chemotherapeutic agent, or molecularly targeted drug are contained as active ingredients in different formulations and administered simultaneously (simultaneously) or at different times (asynchronously or separately). [B26] The anti-EGFR antibody-drug conjugate according to any one of [B1] to [B25], wherein the cancer is selected from the group consisting of lung cancer, colorectal cancer, kidney cancer, liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, cervical cancer, head and neck cancer, bile duct cancer, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule. [B27] The anti-EGFR antibody-drug conjugate according to [B26], wherein the cancer is lung cancer. [B28] The anti-EGFR antibody-drug conjugate according to [B26], wherein the cancer is colorectal cancer. [B29] The anti-EGFR antibody-drug conjugate according to [B26], wherein the cancer is breast cancer. [B30] The anti-EGFR antibody-drug conjugate according to [B26], wherein the cancer is head and neck cancer. [B31] The anti-EGFR antibody-drug conjugate according to [B26], wherein the cancer is renal cancer. [B32] The anti-EGFR antibody-drug conjugate according to [B26], wherein the cancer is liver cancer. [C1] Use of an anti-EGFR antibody-drug conjugate in combination with one or more selected from immune checkpoint inhibitors, chemotherapeutic agents, and molecular targeted drugs in the manufacture of a medicament for treating cancer, wherein the anti-EGFR antibody-drug conjugate is represented by the following formula (I): (In the formula, m ２represents an integer of 1 or 2, L represents a linker connecting the N297 sugar chain and D, -Lb-La-Lp-Lc-* (wherein the asterisk indicates binding to drug D, and Lb is: (In the structural formula of Lb shown above, the asterisk indicates bonding to La, and the wavy line indicates bonding to the N297 sugar chain), where La is -C(=O)-CH ２ CH ２ Lp represents -GGFG- or -GGPI-, Lc represents -NH-CH ２ -), the Ab is an anti-EGFR antibody or an antigen-binding fragment thereof comprising a heavy chain comprising a CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and a light chain comprising a CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, a CDRL2 consisting of the amino acid sequence represented by DAS, and a CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 7, wherein the N297 sugar chain is N297-(Fuc)MSG1 having the structure represented by the following formula: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2nd position of the sialic acid at the non-reducing end of the 1-3 chain side of the branched chain of β-Man in the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1st or 3rd position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), or N297-(Fuc)SG: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２-NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), and D represents (wherein the asterisk indicates binding to L, and W represents -NH-). [C1'] The use according to [C1], wherein Ab is an anti-EGFR antibody comprising an Fc region (wild-type or mutant-type) of the antibody, or an antigen-binding fragment of the antibody. [C2] The use according to [C1], wherein D is (wherein the asterisk indicates binding to L). [C3] The use according to [C1], wherein the anti-EGFR antibody-drug conjugate is represented by the following formula: (Wherein Ab, N297 sugar chain and m ２ [C4] The use according to [C1] or [C2], wherein the anti-EGFR antibody-drug conjugate is represented by the following formula: In the formula, m ２ is 2, the Ab comprises a heavy chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and a light chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2, and the N297 sugar chain is N297-(Fuc)SG having a structure represented by the following formula: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２-NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, the asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring, and n ５[C5] The use according to any one of [C1] to [C4], wherein the Ab comprises a heavy chain variable region consisting of the amino acid sequence shown by amino acid residues 1 to 119 of SEQ ID NO: 1, and a light chain variable region consisting of the amino acid sequence shown by amino acid residues 1 to 107 of SEQ ID NO: 2. [C6] The use according to any one of [C1] to [C5], wherein the Ab comprises a heavy chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence shown by SEQ ID NO: 1, and a light chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence shown by SEQ ID NO: 2. [C7] The use according to any one of [C1] to [C6], wherein the Ab comprises a heavy chain consisting of the amino acid sequence shown by SEQ ID NO: 1 and a light chain consisting of the amino acid sequence shown by SEQ ID NO: 2. [C8] The use according to any one of [C1] to [C5], wherein the Ab comprises an antibody or an antigen-binding fragment of the antibody comprised in the anti-EGFR antibody-drug conjugate according to any one of [C1] to [C7], and the antibody or antigen-binding fragment comprises: (i) a heavy chain consisting of an amino acid sequence in which one or two amino acids are deleted from the carboxyl terminus of the amino acid sequence of the heavy chain; and (ii) a light chain. [C9] The use according to any one of [C1] to [C8], wherein the use of an anti-EGFR antibody-drug conjugate in combination with an immune checkpoint inhibitor in the manufacture of a medicament for treating cancer, wherein the immune checkpoint inhibitor is an antibody selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-TIGIT antibody, and an anti-LAG-3 antibody, an antigen-binding fragment thereof, or a compound comprising any of them. [C10] The use according to [C9], wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, cemiplimab, or dostarlimab. [C11] The use according to [C9], wherein the anti-PD-L1 antibody is atezolizumab, duvalumab, or avelumab. [C12] The use according to [C9], wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab. [C13] The use according to [C9], wherein the anti-TIGIT antibody is tiragolumab or vibostolimab.[C14] The use according to [C9], wherein the anti-LAG-3 antibody is leratolimab. [C15] The use according to any one of [C1] to [C8], wherein the anti-EGFR antibody-drug conjugate is combined with a chemotherapeutic agent in the manufacture of a medicament for the treatment of cancer, wherein the chemotherapeutic agent is an antimetabolite, a tubulin inhibitor, an alkylating agent, a platinum compound, or a topoisomerase inhibitor. [C16] The use according to [C15], wherein the antimetabolite is gemcitabine or a pharmaceutically acceptable salt thereof. [C17] The use according to [C15], wherein the antimetabolite is fluorouracil or a pharmaceutically acceptable salt thereof. [C18] The use according to [C15], wherein the tubulin inhibitor is docetaxel or a pharmaceutically acceptable salt thereof. [C19] The use according to [C15], wherein the platinum compound is oxaliplatin or a pharmaceutically acceptable salt thereof. [C20] The use according to [C15], wherein the platinum agent is cisplatin or a pharmaceutically acceptable salt thereof. [C21] The use according to [C15], wherein the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof. [C22] The use according to any one of [C1] to [C8], wherein an anti-EGFR antibody-drug conjugate is used in combination with a molecularly targeted drug in the manufacture of a medicament for the treatment of cancer, wherein the molecularly targeted drug is a VEGF inhibitor or an EGFR inhibitor. [C23] The use according to [C22], wherein the VEGF inhibitor is bevacizumab, ramucirumab, aflibercept beta, or axitinib. [C24] The use according to [C22], wherein the EGFR inhibitor is osimertinib or a pharmaceutically acceptable salt thereof. [C25] The use according to any one of [C1] to [C24], wherein the anti-EGFR antibody-drug conjugate; and the immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug are contained as active ingredients in separate formulations, and are administered simultaneously (simultaneously) or at different times (asynchronously or separately).[C26] The use according to any one of [C1] to [C25], wherein the cancer is selected from the group consisting of lung cancer, colon cancer, kidney cancer, liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, cervical cancer, head and neck cancer, bile duct cancer, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule. [C27] The use according to [C26], wherein the cancer is lung cancer. [C28] The use according to [C26], wherein the cancer is colon cancer. [C29] The use according to [C26], wherein the cancer is breast cancer. [C30] The use according to [C26], wherein the cancer is head and neck cancer. [C31] The use according to [C26], wherein the cancer is kidney cancer. [C32] The use according to [C26], wherein the cancer is liver cancer. [D1] A method for treating a disease, comprising administering to a subject in need of disease treatment a combination of: a) an anti-EGFR antibody-drug conjugate; and b) one or more selected from an immune checkpoint inhibitor, a chemotherapeutic agent, and a molecular targeted drug, wherein the anti-EGFR antibody-drug conjugate is represented by the following formula (I): (In the formula, m ２ represents an integer of 1 or 2, L represents a linker connecting the N297 sugar chain and D, -Lb-La-Lp-Lc-* (wherein the asterisk indicates binding to drug D, and Lb is: (In the structural formula of Lb shown above, the asterisk indicates bonding to La, and the wavy line indicates bonding to the N297 sugar chain), where La is -C(=O)-CH ２ CH ２ Lp represents -GGFG- or -GGPI-, Lc represents -NH-CH ２-), the Ab is an anti-EGFR antibody or an antigen-binding fragment thereof comprising a heavy chain comprising a CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and a light chain comprising a CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, a CDRL2 consisting of the amino acid sequence represented by DAS, and a CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 7, wherein the N297 sugar chain is N297-(Fuc)MSG1 having the structure represented by the following formula: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of the 1-3 chain side of the branched chain of β-Man in the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1- or 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), or N297-(Fuc)SG: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), and D represents (wherein the asterisk indicates binding to L, and W represents -NH-). [D1'] The method according to [D1], wherein Ab is an anti-EGFR antibody comprising an Fc region (wild-type or mutant-type) of the antibody, or an antigen-binding fragment of the antibody. [D2] The method according to [D1], wherein D is (wherein the asterisk indicates binding to L). [D3] The method according to [D1], wherein the anti-EGFR antibody-drug conjugate is represented by the following formula: (Wherein Ab, N297 sugar chain and m ２ [D4] The method according to [D1] or [D2], wherein the anti-EGFR antibody-drug conjugate is represented by the following formula: In the formula, m ２ is 2, the Ab comprises a heavy chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and a light chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2, and the N297 sugar chain is N297-(Fuc)SG having a structure represented by the following formula: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, the asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring, and n ５[D5] The method of any one of [D1] to [D4], wherein the Ab comprises a heavy chain variable region consisting of the amino acid sequence shown by amino acid residues 1 to 119 of SEQ ID NO: 1, and a light chain variable region consisting of the amino acid sequence shown by amino acid residues 1 to 107 of SEQ ID NO: 2. [D6] The method of any one of [D1] to [D5], wherein the Ab comprises a heavy chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence shown by SEQ ID NO: 1, and a light chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence shown by SEQ ID NO: 2. [D7] The method of any one of [D1] to [D6], wherein the Ab comprises a heavy chain consisting of the amino acid sequence shown by SEQ ID NO: 1 and a light chain consisting of the amino acid sequence shown by SEQ ID NO: 2. [D8] The method of any one of [D1] to [D5], wherein Ab is comprised in an antibody or an antigen-binding fragment of the antibody contained in the anti-EGFR antibody-drug conjugate of any one of [D1] to [D7], and comprises: (i) a heavy chain consisting of an amino acid sequence in which one or two amino acids are deleted from the carboxyl terminus of the amino acid sequence of the heavy chain; and (ii) a light chain. [D9] The method of any one of [D1] to [D8], wherein the method comprises administering in combination to a subject in need of disease treatment a) an anti-EGFR antibody-drug conjugate; and b) an immune checkpoint inhibitor, wherein the immune checkpoint inhibitor is an antibody selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-TIGIT antibody, and an anti-LAG-3 antibody, an antigen-binding fragment thereof, or a compound comprising any of them. [D10] The method according to [D9], wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, cemiplimab, or dostarlimab. [D11] The method according to [D9], wherein the anti-PD-L1 antibody is atezolizumab, duvalumab, or avelumab. [D12] The method according to [D9], wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab.[D13] The method of [D9], wherein the anti-TIGIT antibody is tiragolumab or vibostolimab. [D14] The method of [D9], wherein the anti-LAG-3 antibody is leratolimab. [D15] The method of any one of [D1] to [D8], wherein the method comprises administering in combination to a subject in need of disease treatment a) an anti-EGFR antibody-drug conjugate; and b) a chemotherapeutic agent, wherein the chemotherapeutic agent is an antimetabolite, a tubulin inhibitor, an alkylating agent, a platinum compound, or a topoisomerase inhibitor. [D16] The method of [D15], wherein the antimetabolite is gemcitabine or a pharmaceutically acceptable salt thereof. [D17] The method of [D15], wherein the antimetabolite is fluorouracil or a pharmaceutically acceptable salt thereof. [D18] The method of [D15], wherein the tubulin inhibitor is docetaxel or a pharmaceutically acceptable salt thereof. [D19] The method according to [D15], wherein the platinum agent is oxaliplatin or a pharmaceutically acceptable salt thereof. [D20] The method according to [D15], wherein the platinum agent is cisplatin or a pharmaceutically acceptable salt thereof. [D21] The method according to [D15], wherein the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof. [D22] The method according to any one of [D1] to [D8], wherein the method comprises administering to a subject in need of disease treatment a) an anti-EGFR antibody-drug conjugate; and b) a molecular targeted drug in combination, wherein the molecular targeted drug is a VEGF inhibitor or an EGFR inhibitor. [D23] The method according to [D22], wherein the VEGF inhibitor is bevacizumab, ramucirumab, aflibercept beta, or axitinib. [D24] The method according to [D22], wherein the EGFR inhibitor is osimertinib or a pharmaceutically acceptable salt thereof. [D25] The method according to any one of [D1] to [D24], wherein the anti-EGFR antibody-drug conjugate; and the immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug are contained as active ingredients in separate formulations, and are administered simultaneously (simultaneously) or at different times (asynchronously or separately).[D26] The method according to any one of [D1] to [D25], wherein the disease is cancer. [D27] The method according to [D26], wherein the cancer is selected from the group consisting of lung cancer, colon cancer, kidney cancer, liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, cervical cancer, head and neck cancer, bile duct cancer, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule. [D28] The method according to [D27], wherein the cancer is lung cancer. [D29] The method according to [D27], wherein the cancer is colon cancer. [D30] The method according to [D27], wherein the cancer is breast cancer. [D31] The method according to [D27], wherein the cancer is head and neck cancer. [D32] The method according to [D27], wherein the cancer is kidney cancer. [D33] The method according to [D27], wherein the cancer is liver cancer.

[0011] The present invention provides a novel, safe drug combination therapy that exhibits significant antitumor effects against diseases, particularly cancer.

[0012] Figure 1 shows the amino acid sequence (SEQ ID NO: 1) of the heavy chain of anti-EGFR antibody 1. Figure 2 shows the amino acid sequence (SEQ ID NO: 2) of the light chain of anti-EGFR antibody 1. Figure 3 shows the amino acid sequence of CDRH1 (SEQ ID NO: 3), CDRH2 (SEQ ID NO: 4), CDRH3 (SEQ ID NO: 5), CDRL1 (SEQ ID NO: 6), CDRL2, and CDRL3 (SEQ ID NO: 7) of anti-EGFR antibody 1. Figure 1 shows the antitumor combination effect of intravenous administration of EGFR-ADC (1) and an anti-PD-1 surrogate antibody in mice subcutaneously implanted with EMT6-chimeraEGFR cells, which were prepared by introducing a human-mouse chimeric EGFR gene into the mouse breast cancer cell line EMT6. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the anti-PD-1 surrogate antibody administration group, and the black inverted triangle line indicates the EGFR-ADC(1) and anti-PD-1 surrogate antibody combined administration group. The vertical axis indicates tumor volume (mm ３The horizontal axis indicates the number of days after tumor implantation. The figure shows the antitumor combined effect of intravenous administration of EGFR-ADC(1) and an anti-PD-1 surrogate antibody in mice subcutaneously implanted with CT26.WT-chimeraEGFR cells, a mouse colon cancer cell line CT26.WT introduced with a human mouse chimeric EGFR gene. In the figure, the black circle line indicates the vehicle-administered group, the black square line indicates the EGFR-ADC(1)-administered group, the black triangle line indicates the anti-PD-1 surrogate antibody-administered group, and the black inverted triangle line indicates the group administered with a combination of EGFR-ADC(1) and an anti-PD-1 surrogate antibody. The vertical axis indicates tumor volume (mm ３ The horizontal axis represents the number of days after tumor implantation. This figure shows the antitumor combination effect of intravenous administration of EGFR-ADC(1) and an anti-PD-L1 surrogate antibody in mice subcutaneously implanted with CT26.WT-chimeraEGFR cells, a mouse colon cancer cell line created by introducing a human mouse chimeric EGFR gene into the CT26.WT. In the figure, the black circle line represents the vehicle-administered group, the black square line represents the EGFR-ADC(1)-administered group, the black triangle line represents the anti-PD-L1 surrogate antibody-administered group, and the black inverted triangle line represents the group administered with a combination of EGFR-ADC(1) and an anti-PD-L1 surrogate antibody. The vertical axis represents tumor volume (mm ３ The horizontal axis represents the number of days after tumor implantation. Figure 1 shows the antitumor combined effect of intravenous administration of EGFR-ADC(1) and an anti-CTLA4 surrogate antibody in mice subcutaneously implanted with CT26.WT-chimeraEGFR cells, a mouse colon cancer cell line created by introducing a human mouse chimeric EGFR gene into the CT26.WT. In the figure, the black circle line represents the vehicle-administered group, the black square line represents the EGFR-ADC(1)-administered group, the black triangle line represents the anti-CTLA4 surrogate antibody-administered group, and the black inverted triangle line represents the group administered with a combination of EGFR-ADC(1) and an anti-CTLA4 surrogate antibody. The vertical axis represents tumor volume (mm ３The horizontal axis represents the number of days after tumor implantation. This figure shows the antitumor combined effect of intravenous administration of EGFR-ADC (2) and cisplatin in mice subcutaneously implanted with 4T1-chimeraEGFR cells, which were mouse breast cancer cell line 4T1 introduced with a human-mouse chimeric EGFR gene. In the figure, the black circle line represents the vehicle-administered group, the black square line represents the EGFR-ADC (2)-administered group, the black triangle line represents the cisplatin-administered group, and the black inverted triangle line represents the group administered with a combination of EGFR-ADC (2) and cisplatin. The vertical axis represents tumor volume (mm ３ The horizontal axis represents the number of days after tumor implantation. This figure shows the antitumor combined effect of intravenous administration of EGFR-ADC (2) and oxaliplatin in mice subcutaneously implanted with 4T1-chimeraEGFR cells, which were mouse breast cancer cell line 4T1 introduced with a human mouse chimeric EGFR gene. In the figure, the black circle line represents the vehicle administration group, the black square line represents the EGFR-ADC (2) administration group, the black triangle line represents the oxaliplatin administration group, and the black inverted triangle line represents the EGFR-ADC (2) and oxaliplatin combined administration group. The vertical axis represents tumor volume (mm ３ The horizontal axis represents the number of days after tumor implantation. Figure 1 shows the antitumor effect of intravenous administration of EGFR-ADC(1) and cisplatin in mice subcutaneously implanted with CT26.WT-chimeraEGFR cells, a mouse colon cancer cell line CT26.WT introduced with a human mouse chimeric EGFR gene. In the figure, the black circle line represents the vehicle-administered group, the black square line represents the EGFR-ADC(1)-administered group, the black triangle line represents the cisplatin-administered group, and the black inverted triangle line represents the group administered with a combination of EGFR-ADC(1) and cisplatin. The vertical axis represents tumor volume (mm ３ The horizontal axis represents the number of days after tumor implantation. Figure 1 shows the antitumor effect of intravenous administration of EGFR-ADC(1) and oxaliplatin in mice subcutaneously implanted with CT26.WT-chimeraEGFR cells, a mouse colon cancer cell line CT26.WT introduced with a human mouse chimeric EGFR gene. In the figure, the black circle line represents the vehicle-administered group, the black square line represents the EGFR-ADC(1)-administered group, the black triangle line represents the oxaliplatin-administered group, and the black inverted triangle line represents the group administered with a combination of EGFR-ADC(1) and oxaliplatin. The vertical axis represents tumor volume (mm ３The horizontal axis represents the number of days after tumor implantation. This figure shows the antitumor combined effect of intravenous administration of EGFR-ADC (2) and gemcitabine in mice subcutaneously implanted with 4T1-chimeraEGFR cells, which were mouse breast cancer cell line 4T1 introduced with a human-mouse chimeric EGFR gene. In the figure, the black circle line represents the vehicle-administered group, the black square line represents the EGFR-ADC (2)-administered group, the black triangle line represents the gemcitabine-administered group, and the black inverted triangle line represents the group administered with a combination of EGFR-ADC (2) and gemcitabine. The vertical axis represents tumor volume (mm ３ The horizontal axis represents the number of days after tumor implantation. This figure shows the antitumor combined effect of intravenous administration of EGFR-ADC (2) and fluorouracil in mice subcutaneously implanted with 4T1-chimeraEGFR cells, which were mouse breast cancer cell line 4T1 introduced with a human-mouse chimeric EGFR gene. In the figure, the black circle line represents the vehicle-administered group, the black square line represents the EGFR-ADC (2)-administered group, the black triangle line represents the fluorouracil-administered group, and the black inverted triangle line represents the group administered with a combination of EGFR-ADC (2) and fluorouracil. The vertical axis represents tumor volume (mm ３ The horizontal axis represents the number of days after tumor implantation. This figure shows the antitumor effect of intravenous administration of EGFR-ADC(1) and docetaxel in mice subcutaneously implanted with the human lung adenocarcinoma cell line NCI-H1975 cells. In the figure, the black circle line represents the vehicle-administered group, the black square line represents the EGFR-ADC(1)-administered group, the black triangle line represents the docetaxel-administered group, and the black inverted triangle line represents the EGFR-ADC(1) and docetaxel-administered group. The vertical axis represents tumor volume (mm ３ The horizontal axis indicates the number of days after tumor implantation. Figure 1 shows the antitumor combined effect of intravenous administration of EGFR-ADC (1) and an anti-VEGF surrogate antibody in mice subcutaneously implanted with EMT6-chimeraEGFR cells, which were mouse breast cancer cell line EMT6 introduced with a human-mouse chimeric EGFR gene. In the figure, the black circle line indicates the vehicle-administered group, the black square line indicates the EGFR-ADC (1)-administered group, the black triangle line indicates the anti-VEGF surrogate antibody-administered group, and the black inverted triangle line indicates the combined administration group of EGFR-ADC (1) and an anti-VEGF surrogate antibody. The vertical axis indicates tumor volume (mm ３), the horizontal axis represents the number of days after tumor implantation. This figure shows the antitumor combined effect of intravenous administration of EGFR-ADC (1) and oral administration of osimertinib mesylate in mice subcutaneously implanted with human lung cancer cell line PC-9 cells. In the figure, the black circle line represents the vehicle administration group, the black square line represents the EGFR-ADC (1) administration group, the black triangle line represents the osimertinib mesylate administration group, and the black inverted triangle line represents the EGFR-ADC (1) and osimertinib mesylate combined administration group. The vertical axis represents tumor volume (mm ３ The horizontal axis indicates the number of days after tumor implantation. Figure 1 shows the antitumor effect of intravenously administered EGFR-ADC(2), an anti-PD-1 surrogate antibody, and cisplatin in mice subcutaneously implanted with LL / 2-chimeraEGFR cells, which were prepared by introducing a human-mouse chimeric EGFR gene into the mouse lung cancer cell line LL / 2. In the figure, the black circle line represents the vehicle administration group, the black square line represents the EGFR-ADC (2) administration group, the black triangle line represents the anti-PD-1 surrogate antibody administration group, the black inverted triangle line represents the cisplatin administration group, the black diamond line represents the EGFR-ADC (2) and anti-PD-1 surrogate antibody combined administration group, the white circle line represents the EGFR-ADC (2) and cisplatin combined administration group, the white square line represents the anti-PD-1 surrogate antibody and cisplatin combined administration group, and the white triangle line represents the EGFR-ADC (2), anti-PD-1 surrogate antibody and cisplatin combined administration group. The vertical axis represents tumor volume (mm ３ The horizontal axis indicates the number of days after tumor inoculation. The mean tumor volume (mm ) on each day after tumor inoculation in each administration group in the test shown in Figure 17 ３ ) are shown. The figure shows the antitumor combined effect of intravenous administration of EGFR-ADC (1) and intraperitoneal administration of irinotecan hydrochloride hydrate in mice subcutaneously implanted with human lung adenocarcinoma cell line NCI-H1975 cells. In the figure, the black circle line represents the vehicle administration group, the black square line represents the EGFR-ADC (1) administration group, the black triangle line represents the irinotecan hydrochloride hydrate administration group, and the black inverted triangle line represents the group administered with a combination of EGFR-ADC (1) and irinotecan hydrochloride hydrate. The vertical axis represents tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation.

[0013] Preferred embodiments for carrying out the present invention will be described below. Note that the embodiment described below shows one example of a typical embodiment of the present invention, and the scope of the present invention should not be construed as being narrow.

[0014] 1. Anti-EGFR Antibody-Drug Conjugate The anti-EGFR antibody-drug conjugate used in the present invention has the following formula (I): It is expressed as:

[0015] Ab represents an anti-EGFR antibody or an antigen-binding fragment of the antibody, N297 sugar chain represents an N-linked sugar chain bound to the side chain of Asn 297 of the IgG heavy chain contained in Ab, L represents a linker connecting the N297 sugar chain and D, D represents a drug that is a cyclic dinucleotide derivative, and m ２ represents an integer of 1 or 2.

[0016] 1-1. Anti-EGFR Antibody The anti-EGFR antibody in the antibody-drug conjugate used in the present invention may be derived from any species, but is preferably an antibody derived from human, rat, mouse, or rabbit. When the antibody is derived from a species other than human, it is preferably chimerized or humanized using well-known techniques. The anti-EGFR antibody in the antibody-drug conjugate used in the present invention may be a polyclonal antibody or a monoclonal antibody, but is preferably a monoclonal antibody.

[0017] It is known that the heavy and light chains of an antibody molecule each contain three complementarity determining regions (CDRs). CDRs, also known as hypervariable regions, are located within the variable regions of the heavy and light chains of an antibody and are particularly highly variable in their primary structure. They are separated into three regions in the primary structure of the heavy and light chain polypeptide chains. In this specification, the CDRs of an antibody are represented as CDRH1, CDRH2, and CDRH3 from the amino-terminus of the heavy chain amino acid sequence, and CDRL1, CDRL2, and CDRL3 from the amino-terminus of the light chain amino acid sequence. These regions are close to each other in the three-dimensional structure and determine the specificity for the antigen to which they bind. In the present invention, the positions and lengths of CDRs were determined according to the IMGT definition (Developmental and Comparative Immunology 27 (2003) 55-77).

[0018] It is known that there are multiple allotypes of antibody constant regions. For example, for IgG1 heavy chains, allotypes include G1m17, G1m3, G1m1, and G1m2. The antibody constant region used in the present invention is not particularly limited, but it is preferable to use G1m17 or G1m3.

[0019] In the present invention, the term "antigen-binding fragment of an anti-EGFR antibody" refers to an antibody fragment having antigen-binding activity, and has a well-conserved N-linked glycan (referred to as "Asn297 glycan" or "N297 glycan" in the present specification and claims) at the 297th asparagine residue (hereinafter referred to as "Asn297 or N297") in the Fc region of the IgG heavy chain. Furthermore, these antigen-binding fragments include not only those obtained by treating the full-length antibody protein molecule with an appropriate enzyme, but also proteins produced in appropriate host cells using genetically engineered antibody genes.

[0020] The "anti-EGFR antibody" in the antibody-drug conjugate used in the present invention is an antibody that specifically binds to EGFR (Epidermal Growth Factor Receptor; ERBB; HER1), and preferably has the activity of being internalized into EGFR-expressing cells upon binding to EGFR.

[0021] Examples of anti-EGFR antibodies include panitumumab, cetuximab, necitumumab, matuzumab, nimotuzumab, zalutumumab, ametumumab (SY-101), SYN-004, SCT-200, tomuzotuximab, GC-1118, GR-1401, depatuxizumab (ABT-806), serculutamab, and AMG595, and preferably panitumumab.

[0022] Substitution of some amino acid residues in the constant region can adjust effector function (see WO88 / 07089, WO94 / 28027, WO94 / 29351). An example of an IgG1 mutant is the IgG1 LALA mutation (IgG1-L234A, L235A). L234A and L235A represent substitutions of leucine with alanine at positions 234 and 235, as defined by the EU index (Proceedings of the National Academy of Sciences of the United States of America, Vol. 63, No. 1 (May 15, 1969), pp. 78-85).

[0023] Examples of anti-EGFR antibodies include antibodies whose heavy chain constant region is that of human IgG1, in which the leucine at position 234, the leucine at position 235, and the aspartic acid at position 265, as shown in the EU index, are substituted with alanine, alanine, and glycine, respectively. More preferred examples of anti-EGFR antibodies include antibodies which contain a heavy chain constant region of human IgG1 having the above amino acid substitutions, and which include a heavy chain comprising a CDRH1 consisting of the amino acid sequence represented by SEQ ID NO:3, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO:4, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO:5, and a CDRL1 consisting of the amino acid sequence represented by SEQ ID NO:6, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO:7, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO:8. and a light chain comprising a CDRL2 consisting of the amino acid sequence set forth in SEQ ID NO:7, and a CDRL3 consisting of the amino acid sequence set forth in SEQ ID NO:7; more preferably, an antibody comprising a heavy chain constant region of human IgG1 having the amino acid substitution, and comprising a heavy chain comprising a heavy chain variable region consisting of the amino acid sequence set forth in amino acid residues 1 to 119 of SEQ ID NO:1, and a light chain comprising a light chain variable region consisting of the amino acid sequence set forth in amino acid residues 1 to 107 of SEQ ID NO:2; and even more preferably, an antibody comprising a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:1 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:2.

[0024] Furthermore, it is known that when antibodies are produced in cultured mammalian cells, the lysine residue at the carboxyl terminus of the heavy chain is deleted (Journal of Chromatography A, 705:129-134 (1995)), and that two amino acid residues, glycine and lysine, are deleted from the carboxyl terminus of the heavy chain, and a proline residue newly positioned at the carboxyl terminus is amidated (Analytical Biochemistry, 360:75-83 (2007)). Thus, the antibodies of the present invention also include deletion antibodies in which one or two amino acids are deleted from the carboxy terminus of the heavy chain, as well as amidated versions of such deletion antibodies (for example, antibodies in which the proline residue at the carboxyl terminus of the heavy chain is amidated).

[0025] In the present invention, examples of the heavy and light chains contained in an anti-EGFR antibody, i.e., an antibody that binds to EGFR, or an antigen-binding fragment of the antibody include the following (i) to (vi): (i) (i-H) a heavy chain comprising a CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and (i-L) a light chain comprising a CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, a CDRL2 consisting of the amino acid sequence represented by DAS, and a CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 7; (ii) (i) A combination of heavy and light chains according to (ii-H), wherein the heavy chain comprises a heavy chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and (ii-L) a light chain comprising a light chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2; (iii) A combination of heavy and light chains according to (i) or (ii), wherein (iii-H) the heavy chain comprises a heavy chain variable region consisting of the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and (iii-L) the light chain comprises a light chain variable region consisting of the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2; (iv) (iv-H) a heavy chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence shown in SEQ ID NO: 1, and (iv-L) a light chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence shown in SEQ ID NO: 2; (v) a heavy chain and light chain combination according to (i), (ii), (iii) or (iv), wherein the heavy chain and the light chain comprise a heavy chain constant region of human IgG1 in which leucine at position 234, leucine at position 235, and aspartic acid at position 265, as defined by the EU index, are substituted with alanine, alanine, and glycine, respectively;(vi) A combination of a heavy chain and a light chain according to (i), (ii), (iii), (iv), or (v), wherein (v-H) a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 1, and (v-L) a light chain consisting of the amino acid sequence set forth in SEQ ID NO: 2; (vii) a heavy chain consisting of an amino acid sequence in which one or two amino acids are deleted from the carboxyl terminus of the amino acid sequence of the heavy chain (heavy chain a) set forth in any one of (i-H) to (v-H), and a light chain (light chain a) combined with heavy chain a in (i) to (v); Examples include, but are not limited to, preferred examples are (i) to (vii), and more preferred examples are (vi) and (vii). The anti-EGFR antibody or antigen-binding fragment thereof of the present invention preferably comprises an antibody Fc region (wild-type or mutant), more preferably an IgG-derived Fc region (wild-type or mutant), and even more preferably an IgG-derived Fc region (wild-type or mutant). Mutant Fc regions may be naturally occurring or artificially created, and preferably include those in which the leucine at position 234, the leucine at position 235, and the aspartic acid at position 265, as determined by the EU index, are substituted with alanine, alanine, and glycine, respectively.

[0026] In the above (ii) or (iv), the numerical values ​​"80%, " "90%" or "95%" are not limited to these and may be, for example, any of 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, and may preferably be any of the numerical values ​​of 90% or more, and more preferably be any of the numerical values ​​of 95% or more.

[0027] 1-2. Antibody N297 Glycan Chain IgG has a well-conserved N-linked glycan (hereinafter referred to as "Asn297 glycan" or "N297 glycan") at the 297th asparagine residue (hereinafter referred to as "Asn297" or "N297") in the Fc region of its heavy chain, and is known to contribute to the activity, kinetics, etc. of antibody molecules (Eon-Duval, A. et al., Biotechnol. Prog. 2012, 28, 608-622; Sanglier-Cianferani, S., Anal. Chem. 2013, 85, 715-736).

[0028] The amino acid sequence in the constant region of IgG is well conserved, and in a report by Edelman et al. (Proc. Natl. Acad. Sci. U.S.A., 63, 78-85, (1969)), each amino acid is identified by an EU number (EU INDEX). For example, Asn297, to which an N-linked sugar chain is added in the Fc region, corresponds to position 297 in the EU numbering. Even if the actual amino acid position changes due to molecular fragmentation or region deletion, the amino acid can be uniquely identified by displaying it by EU numbering.

[0029] The N297 sugar chain used in the antibody-drug conjugate of the present invention is N297-(Fuc)SG or N297-(Fuc)MSG1, preferably N297-(Fuc)SG.

[0030] N297-(Fuc)SG is represented by the following structural formula or sequence formula:

[0031]

[0032]

[0033] In the above formula, the wavy line indicates binding to Asn297 of the antibody, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２-NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, and the asterisk indicates that it is bonded to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring of Lb in the linker L, in particular, ５ is an integer of 2 to 5, preferably 3.

[0034] N297-(Fuc)MSG1 is represented by the following structural formula or sequence formula:

[0035]

[0036]

[0037] In the above formula, the wavy line indicates binding to Asn297 of the antibody, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, which indicates that the amino group at the right end of L (PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of the 1-3 chain side of the branched chain of β-Man in the N297 sugar chain; an asterisk indicates that the linker L, particularly the linker L, is bonded to the nitrogen atom at the 1- or 3-position on the 1,2,3-triazole ring of Lb; ５ is an integer of 2 to 5, preferably 3.

[0038] When the N297 sugar chain of the antibody in the antibody-drug conjugate of the present invention is N297-(Fuc)SG, the antibody-drug conjugate is a molecule to which four linkers L and four drugs D are bound (the above m ２ = 2).

[0039] When the N297 sugar chain of the antibody in the antibody-drug conjugate of the present invention is N297-(Fuc)MSG1, the antibody-drug conjugate is a molecule to which two linkers L and two drugs D are bound (the above m ２ = 1).

[0040] When the N297 sugar chain of the antibody in the antibody-drug conjugate of the present invention is N297-(Fuc)SG or N297-(Fuc)MSG1, a highly homogeneous ADC can be obtained.

[0041] 1-3. Drug The drug of the anti-EGFR antibody-drug conjugate used in the present invention (D in formula (I) above, also referred to as drug D) is (wherein the asterisk indicates bonding to L, and W represents —NH—).

[0042] Preferably, and more preferably, (where the asterisk indicates binding to L).

[0043] 1-4. Linker The linker connecting the N297 sugar chain and drug D is represented by -Lb-La-Lp-Lc-*, where the asterisk indicates binding to drug D, Lp is -GGGFG- or -GGPI-, preferably -GGGFG-, and La is -C(=O)-CH ２ CH ２ Lb represents —C(═O)—; (In the structural formula of Lb shown above, the asterisk indicates bonding to La, and the wavy line indicates bonding to the N297 sugar chain), and Lc represents -NH-CH ２ Indicates -.

[0044] In the present invention, a drug linker refers to a compound in which a drug D in an antibody-drug conjugate is bound to a linker moiety, and a drug linker of the following formula x1, x2, y1, or y2 is preferably used, and a drug linker of formula x1 or x2 is more preferably used. (In the above formulae x1, x2, y1 and y2, the wavy lines indicate binding to the N297 sugar chain).

[0045] 2. Production of anti-EGFR antibody-drug conjugates 2-1. Production of antibodies The anti-EGFR antibodies described in 1-1 can be obtained by known means (e.g., Proc. Natl. Acad. Sci. U.S.A., 81, 6851-6855, (1984), Nature (1986) 321, pp. 522-525, WO90 / 07861, WO1998 / 050433, WO2002 / 092771).

[0046] Recently, a method has been reported in which heterogeneous antibody sugar chains are remodeled (modified) by enzymatic reaction to uniformly introduce sugar chains bearing functional groups (ACS Chem. Biol. 2012, 7, 110-122, ACS Med. Chem. Lett. 2016, 7, 1005-1008). Attempts have also been made to use this sugar chain remodeling technique to site-specifically introduce drugs and synthesize homogeneous ADCs (Bioconjugate Chem. 2015, 26, 2233-2242, Angew. Chem. Int. Ed. 2016, 55, 2361-2367, US2016361436).

[0047] The antibody having an N297 sugar chain used in the present invention can be produced by applying sugar chain remodeling technology to the antibody described in 1-1., for example, in accordance with the methods described in WO2018 / 003983, WO2020 / 050406, WO2021 / 177438, WO2022 / 050300, WO2023 / 167238, PLos ONE 2018, 13, e0193534, etc.

[0048] 2-2. Production of Drugs and Linkers The drugs and linkers used in the present invention can be produced with reference to the descriptions in WO 2020 / 050406, WO 2021 / 177438, WO 2022 / 163846, and the like.

[0049] 2-3. Production of Drug Linker Intermediate (Conjugation Precursor) The drug linker intermediate (conjugation precursor) used in the production of the anti-EGFR antibody-drug conjugate used in the present invention is represented by the following formula:

[0050] More preferably, a drug linker intermediate (conjugation precursor) represented by the following formula is used:

[0051] The above drug linker intermediate (conjugation precursor) can be produced with reference to the descriptions in WO 2020 / 050406, WO 2021 / 177438, WO 2022 / 163846, and the like. The compound of formula (Rp, Rp-X) in WO 2022 / 163846 can be produced by deprotecting the protecting group of the compound of formula (Rp, Rp-VII′), which is the precursor of that compound, using tetrabutylammonium fluoride (TBAF). In this production process, a silicon reagent (e.g., TBSO-CH ) can be used as a quenching agent for TBAF instead of a base (e.g., calcium chloride, calcium carbonate). ２ CH ２ —OH) can be used.

[0052] 2-4. Production of Anti-EGFR Antibody-Drug Conjugates The anti-EGFR antibody-drug conjugates used in the present invention can be produced by linking the aforementioned drug linker intermediate (conjugation precursor) with an antibody having an N297 sugar chain via a cycloaddition reaction. Examples of cycloaddition reactions include the Diels-Alder reaction and the 1,3-dipolar cycloaddition reaction, preferably the 1,3-dipolar cycloaddition reaction. Examples of 1,3-dipolar cycloaddition reactions include the cycloaddition reaction of an azide with a terminal alkyne and the SPAAC (strain-promoted azide-alkyne cycloaddition: J. Am. Chem. Soc. 2004, 126, 15046-15047) reaction, preferably the SPAAC reaction.

[0053] The above anti-EGFR antibody-drug conjugate can be produced with reference to the descriptions in WO 2020 / 050406, WO 2021 / 177438, WO 2022 / 163846, and the like.

[0054] 3. Immune Checkpoint Inhibitors In the present invention, the term "immune checkpoint inhibitor" refers to a drug that inhibits the immunosuppressive system and activates tumor immunity. The immune checkpoint inhibitor used in the present invention is not particularly limited, but preferred examples include anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA-4 antibodies, anti-TIGIT antibodies, and anti-LAG-3 antibodies, and more preferred examples include anti-PD-1 antibodies, anti-PD-L1 antibodies, and anti-CTLA-4 antibodies.

[0055] In the present invention, the term "anti-PD-1 antibody" refers to an antibody that specifically binds to PD-1 (programmed cell death-1; CD279; PDCD1), and preferably refers to an antibody that has the effect of reducing, inhibiting, and / or interfering with signal transduction resulting from the interaction between PD-1 and its binding partners, PD-L1 and PD-L2. The anti-PD-1 antibody used in the present invention is not particularly limited as long as its clinical efficacy and safety have been confirmed. Preferred examples include nivolumab (WO 2006 / 121168, etc.), pembrolizumab (WO 2008 / 156712, etc.), dostarlimab, spartalizumab (WO 2015 / 112900, etc.), and cemiplimab (WO 2015 / 196051, etc.). Furthermore, for the purpose of confirming the combined effect of the antibody-drug conjugate used in the present invention in preclinical studies, commercially available anti-PD-1 antibodies for research use (e.g., clone RMP1-14, Anti-mPD-1 mIgG1e3), etc. These anti-PD-1 antibodies are commonly used as surrogate antibodies that inhibit PD-1 signaling in mice.

[0056] In the present invention, the term "anti-PD-L1 antibody" refers to an antibody that specifically binds to PD-L1 (Programmed cell death ligand 1; CD274; B7-H1), and preferably refers to an antibody that has the effect of reducing, inhibiting, and / or interfering with signal transduction resulting from the interaction between PD-L1 and its binding partners, PD-1 and B7.1 (CD80). The anti-PD-L1 antibody used in the present invention is not particularly limited as long as its clinical efficacy and safety have been confirmed, but preferred examples include atezolizumab (WO 2010 / 077634, etc.), durvalumab (WO 2011 / 066389, etc.), and avelumab (WO 2013 / 079174, etc.). Furthermore, for the purpose of confirming the combined effect of the antibody-drug conjugate used in the present invention in preclinical studies, commercially available anti-PD-L1 antibodies for research use (e.g., clone 10F.9G2) can also be used. These anti-PD-L1 antibodies are commonly used as surrogate antibodies that inhibit PD-L1 signaling in mice.

[0057] In the present invention, the term "anti-CTLA-4 antibody" refers to an antibody that specifically binds to CTLA-4 (cytotoxic T-lymphocyte-associated protein 4; CD152), and preferably refers to an antibody that has the effect of reducing, inhibiting, and / or interfering with signal transduction resulting from the interaction between CTLA-4 and its binding partners, B7.1 (CD80) and B7.2 (CD86). The anti-CTLA-4 antibody used in the present invention is not particularly limited as long as its clinical efficacy and safety have been confirmed, but preferred examples include ipilimumab (WO 2001 / 014424, etc.) and tremelimumab (WO 2000 / 037504, etc.). Furthermore, for the purpose of confirming the combined effect of the anti-EGFR antibody-drug conjugate used in the present invention in preclinical studies, commercially available anti-CTLA-4 antibodies for research use (e.g., clones 9D9 and 9H10) can also be used. These anti-CTLA-4 antibodies are commonly used as surrogate antibodies that inhibit CTLA-4 signaling in mice.

[0058] In the present invention, the term "anti-TIGIT antibody" refers to an antibody that specifically binds to TIGIT (T cell immunoreceptor with Ig and ITIM domains), and preferably refers to an antibody that has the effect of reducing, inhibiting, and / or interfering with signal transduction resulting from the interaction between TIGIT and its binding partner, CD155 or the like. The anti-TIGIT antibody used in the present invention is not particularly limited as long as its clinical efficacy and safety have been confirmed, but preferred examples include tiragolumab (WO 2017 / 053748) and vibostolimab (WO 2016 / 028656). Furthermore, for the purpose of confirming the combined effect of the anti-EGFR antibody-drug conjugate used in the present invention in preclinical studies, commercially available anti-TIGIT antibodies for research use (e.g., clone 1G9) and the like can also be used.

[0059] In the present invention, the term "anti-LAG-3 antibody" refers to an antibody that specifically binds to LAG-3 (lymphocyte-activation gene 3), and preferably refers to an antibody that has the effect of reducing, inhibiting, and / or interfering with signal transduction resulting from the interaction between LAG-3 and its binding partner, major histocompatibility complex (MHC) class II. The anti-LAG-3 antibody used in the present invention is not particularly limited as long as its clinical efficacy and safety have been confirmed, and a preferred example is relatlimab. Furthermore, for the purpose of confirming the combined effect of the anti-EGFR antibody-drug conjugate used in the present invention in preclinical studies, commercially available research-grade anti-LAG-3 antibodies (e.g., clone C9B7W) and the like can also be used.

[0060] In the present invention, the immune checkpoint inhibitor used in combination with the anti-EGFR antibody-drug conjugate may be an antigen-binding fragment of any of the above antibodies, or a compound containing the antibody or the antigen-binding fragment. Here, the antigen-binding fragment refers to a fragment of an antibody that has antigen-binding activity, and includes Fab, F(ab')2, Fv, scFv, diabody, linear antibody, and multispecific antibody formed from antibody fragments. However, the antigen-binding fragment is not limited to these molecules as long as it has antigen-binding ability.

[0061] 4. Chemotherapeutic Agents In the present invention, the term "chemotherapeutic agent" refers to a chemically synthesized drug among compounds having anticancer or antitumor activity. The chemotherapeutic agent used in the present invention is not particularly limited, but suitable examples include topoisomerase inhibitors, tubulin inhibitors, antimetabolites, platinum compounds, DNA demethylating agents, anticancer antibiotics, and alkylating agents, and may also be pharmaceutically acceptable salts of these drugs.

[0062] Examples of topoisomerase inhibitors include irinotecan, topotecan, etoposide, etc., and irinotecan is preferred.

[0063] Examples of tubulin inhibitors include paclitaxel, docetaxel, vincristine, vinblastine, vindesine, eribulin, vinorelbine, albumin-suspended paclitaxel (Nab-paclitaxel), and the like, with docetaxel being preferred.

[0064] Examples of antimetabolites include fluorouracil (also called 5-FU), mercaptopurine, decitabine, gemcitabine (also called GEM), azacitidine (also called 5-Aza), methotrexate, tegafur, UFT, S-1, carmofur, doxifluridine, and capecitabine, and preferred examples include fluorouracil and gemcitabine.

[0065] Examples of platinum preparations include oxaliplatin, carboplatin, cisplatin, nedaplatin, etc., and preferred examples include oxaliplatin and cisplatin.

[0066] Examples of DNA demethylating agents include azacytidine and decitabine.

[0067] Examples of anticancer antibiotics include doxorubicin, bleomycin, and liposomal doxorubicin.

[0068] Examples of the alkylating agent include ifosfamide, cyclophosphamide, dacarbazine, etc., and preferably cyclophosphamide.

[0069] However, the chemotherapeutic agents that can be used in combination with the anti-EGFR antibody-drug conjugates of the present invention are not limited to these.

[0070] 7. Molecularly Targeted Drugs In the present invention, the term "molecularly targeted drug" refers to a drug designed to attack genes or proteins present in or around cancer cells and particularly involved in cancer cell proliferation, with the main targets being cell surface antigens, signaling molecules, ligands, and the like. The molecularly targeted drug used in the present invention is not particularly limited, but preferred examples include angiogenesis inhibitors and kinase inhibitors. More preferred examples of angiogenesis inhibitors include VEGF inhibitors, and more preferred examples of kinase inhibitors include EGFR inhibitors, FGFR inhibitors, and multikinase inhibitors, and pharmaceutically acceptable salts of these drugs may also be used. Examples of VEGF inhibitors include bevacizumab, ramucirumab, aflibercept beta, and axitinib. Examples of EGFR inhibitors include gefitinib, erlotinib, afatinib, osimertinib, cetuximab, panitumumab, necitumumab, matuzumab, nimotuzumab, zalutumumab, etc., with osimertinib being preferred. Preferred examples of FGFR inhibitors include pemigatinib, futibatinib, etc. Preferred examples of multikinase inhibitors include sorafenib, sunitinib, pazopanib, regorafenib, lenvatinib, etc.

[0071] 8. Pharmaceutical compositions, treatment methods, etc.

[0072] Hereinafter, the pharmaceutical composition used in the combination therapy of the anti-EGFR antibody-drug conjugate according to the present invention with an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecular targeted drug, and the combination therapy (treatment method) will be described.

[0073] In the present invention, "used in combination" means that a recipient takes in multiple drugs into their body over a certain period of time. Furthermore, multiple drugs do not need to be present in the body at the same time; they may be taken into the body over a certain period of time (e.g., one month, preferably one week, more preferably several days, and even more preferably one day), and the other active ingredient may have disappeared from the body by the time one of them is administered. In the present invention, the anti-EGFR antibody-drug conjugate and the immune checkpoint inhibitor, chemotherapeutic agent, or molecularly targeted drug may each be contained as active ingredients in separate formulations. In such cases, the timing and number of administrations are not particularly limited. For example, they may be administered simultaneously (simultaneously) or at different times (asynchronously or separately), preferably simultaneously. When administered at different times, they may be administered consecutively or discontinuously. When administered at different times, the administration interval and order can be changed as appropriate. Furthermore, in the present invention, the anti-EGFR antibody-drug conjugate and the immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug may be contained as active ingredients in a single preparation.

[0074] The present invention also encompasses embodiments in which an anti-EGFR antibody-drug conjugate is used in combination with two or more drugs selected from immune checkpoint inhibitors, chemotherapeutic agents, and molecularly targeted drugs. In this case, the two or more drugs may be selected from drugs belonging to the same category (e.g., two drugs belonging to immune checkpoint inhibitors) or from drugs belonging to different categories (e.g., a drug belonging to immune checkpoint inhibitors and a drug belonging to chemotherapeutic agents). For example, in one embodiment of the present invention, a pharmaceutical composition comprising an anti-EGFR antibody-drug conjugate can be used in combination with two or more drugs selected from immune checkpoint inhibitors, chemotherapeutic agents, and molecularly targeted drugs. Furthermore, a pharmaceutical composition comprising an immune checkpoint inhibitor, chemotherapeutic agent, or molecularly targeted drug can be used in combination with the anti-EGFR antibody-drug conjugate and one or more other drugs selected from immune checkpoint inhibitors, chemotherapeutic agents, and molecularly targeted drugs. In another aspect of the present invention, a pharmaceutical composition comprising an anti-EGFR antibody-drug conjugate further comprises two or more drugs selected from an immune checkpoint inhibitor, a chemotherapeutic agent, and a molecularly targeted drug; and in yet another aspect of the present invention, a pharmaceutical composition comprising (i) an anti-EGFR antibody-drug conjugate and (ii) an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug may be used in combination with one or more additional drugs selected from an immune checkpoint inhibitor, a chemotherapeutic agent, and a molecularly targeted drug.

[0075] The pharmaceutical composition and treatment method of the present invention can be used to treat cancer, preferably at least one cancer selected from the group consisting of lung cancer, kidney cancer, urothelial cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, hepatocellular carcinoma, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, testicular cancer, cervical cancer, placental choriocarcinoma, brain tumor, head and neck cancer, thyroid cancer, mesothelioma, gastrointestinal stromal tumor (GIST), gallbladder cancer, bile duct cancer, adrenal cancer, squamous cell carcinoma, pharyngeal cancer, tongue cancer, ear cancer, thymic cancer, small intestine cancer, leukemia, malignant lymphoma, plasmacytoma, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule. Renal cancer includes renal cell carcinoma. Lung cancer includes non-small cell lung cancer.

[0076] EGFR mutation-positive cancer is cancer with a mutation in EGFR. Examples of EGFR mutation-positive cancer include lung cancer and head and neck cancer that contain at least one mutation in exons 18 to 21, and preferably lung cancer with an exon 19 mutation.

[0077] Cancers with a mutation in an EGFR downstream signaling molecule are cancers with a mutation in a downstream molecule of the EGFR signaling pathway, and examples of cancers with a mutation in an EGFR downstream signaling molecule include cancers with a mutation in at least one of RAS, RAF, PI3K, etc., preferably RAS-mutated cancers such as lung cancer, colon cancer, and pancreatic cancer with a mutation in at least one of RAS (KRAS, NRAS, HRAS), more preferably lung cancer or colon cancer with a KRAS mutation, and even more preferably lung cancer with a KRAS mutation.

[0078] In another aspect, the pharmaceutical composition and treatment method of the present invention can be used for the treatment of at least one cancer selected from the group consisting of lung cancer, colorectal cancer, kidney cancer, liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, cervical cancer, head and neck cancer, bile duct cancer, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule.

[0079] In another embodiment, the pharmaceutical compositions and treatment methods of the present invention can be used to treat lung cancer.

[0080] In another embodiment, the pharmaceutical compositions and treatment methods of the present invention can be used to treat breast cancer.

[0081] In another embodiment, the pharmaceutical compositions and treatment methods of the present invention can be used to treat head and neck cancer.

[0082] In another embodiment, the pharmaceutical compositions and treatment methods of the present invention can be used to treat colon cancer.

[0083] In another embodiment, the pharmaceutical compositions and treatment methods of the present invention can be used to treat renal cancer.

[0084] In another embodiment, the pharmaceutical composition and treatment method of the present invention can be used to treat liver cancer.

[0085] In one aspect, the present invention relates to a pharmaceutical composition comprising (i) an anti-EGFR antibody-drug conjugate and (ii) an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug.

[0086] In one aspect, the present invention relates to an anti-EGFR antibody-drug conjugate for use in combination with an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug for the treatment of cancer.

[0087] In one aspect, the present invention relates to an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug for use in combination with an anti-EGFR antibody-drug conjugate for the treatment of cancer.

[0088] In one aspect, the present invention relates to the use of an anti-EGFR antibody-drug conjugate in combination with an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug in the manufacture of a medicament for the treatment of cancer. In another aspect of the present invention, the present invention relates to the use of an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug in combination with an anti-EGFR antibody-drug conjugate in the manufacture of a medicament for the treatment of cancer. In another aspect of the present invention, the present invention relates to the use of an anti-EGFR antibody-drug conjugate and an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug in the manufacture of a medicament for the combination treatment of cancer; the use of an anti-EGFR antibody-drug conjugate in the manufacture of a cancer therapeutic agent in combination with an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug; or the use of an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug in the manufacture of a cancer therapeutic agent in combination with an anti-EGFR antibody-drug conjugate.

[0089] In one aspect, the present invention relates to a method for treating a disease, comprising administering to a subject in need thereof (i) an anti-EGFR antibody-drug conjugate and (ii) an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug in combination.

[0090] In one aspect, the present invention relates to a pharmaceutical composition for the treatment of cancer comprising an anti-EGFR antibody-drug conjugate for use in combination with an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug. In another aspect, the present invention relates to a pharmaceutical composition for the treatment of cancer comprising an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug for use in combination with an anti-EGFR antibody-drug conjugate.

[0091] In one aspect, the present invention relates to a cancer therapeutic agent which is used in combination with an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug, and which is intended for patients undergoing treatment with the immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug, and which comprises an anti-EGFR antibody-drug conjugate as an active ingredient. In another aspect, the present invention relates to a cancer therapeutic agent which is used in combination with an anti-EGFR antibody-drug conjugate, and which comprises an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug as an active ingredient, and which is intended for patients undergoing treatment with the anti-EGFR antibody-drug conjugate.

[0092] In one aspect, the present invention relates to a cancer therapeutic agent for use in the treatment of cancer, which comprises an anti-EGFR antibody-drug conjugate as an active ingredient, wherein the treatment comprises the administration of an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug in addition to the anti-EGFR antibody-drug conjugate. In another aspect, the present invention relates to a cancer therapeutic agent for use in the treatment of cancer, which comprises an immune checkpoint inhibitor, a chemotherapeutic agent, or a molecularly targeted drug as an active ingredient, wherein the treatment comprises the administration of an anti-EGFR antibody-drug conjugate in addition to the immune checkpoint inhibitor, the chemotherapeutic agent, or the molecularly targeted drug.

[0093] The pharmaceutical composition and treatment method of the present invention can be suitably used when EGFR expression is confirmed in cancer.

[0094] EGFR expression can be confirmed, for example, by detection at the level of the EGFR gene product (protein) using immunohistochemistry (IHC), a flow cytometer, Western blot analysis, or the like, or by detection at the gene transcription level using in situ hybridization (ISH) or quantitative PCR (q-PCR).

[0095] As used herein, a "subject" or "patient" refers to a human or other animal in need of such treatment, and in one embodiment, a human in need of such treatment.

[0096] The antitumor effects of the pharmaceutical composition and treatment method of the present invention can be confirmed, for example, by preparing a model in which cancer cells are transplanted into a test animal and measuring the reduction in tumor volume and the life-prolonging effect of administering the pharmaceutical composition and treatment method of the present invention.The combined effect of (i) the anti-EGFR antibody-drug conjugate and (ii) the immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug used in the present invention can be confirmed by comparing the antitumor effects with those of the anti-EGFR antibody-drug conjugate, immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug used in the present invention when administered alone.

[0097] The antitumor effects of the pharmaceutical composition and treatment method of the present invention can be confirmed in clinical trials by the Response Evaluation Criteria in Solid Tumors (RECIST) evaluation method, the WHO evaluation method, the Macdonald evaluation method, weight measurement, and other methods, and the results can be evaluated for complete response (CR), partial response (PR), progressive disease (PD), objective response rate (ORR), duration of response (DoR), progression-free survival (PFS), overall survival (OS), and other important parameters. The survival rate can be determined by an index such as OS (Optical Survival Rate).

[0098] By the above-mentioned method, it is possible to confirm the superiority of the antitumor effect of the pharmaceutical composition and treatment method of the present invention over existing pharmaceutical compositions and treatment methods for cancer treatment.

[0099] The pharmaceutical composition and treatment method of the present invention can suppress the proliferation of cancer cells, thereby enabling cancer patients to achieve a higher quality of life and longer survival.

[0100] The pharmaceutical composition of the present invention can be administered containing one or more pharmaceutically compatible ingredients. The pharmaceutically compatible ingredients can be appropriately selected from formulation additives and other ingredients commonly used in this field depending on the dosage and administration concentration of the anti-EGFR antibody-drug conjugate, immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug used in the present invention.

[0101] The pharmaceutical composition and treatment method of the present invention may further comprise a cancer therapeutic agent other than the anti-EGFR antibody-drug conjugate, immune checkpoint inhibitor, chemotherapeutic agent, or molecularly targeted drug of the present invention. The pharmaceutical composition of the present invention can also be administered in combination with other cancer therapeutic agents, thereby enhancing the anti-tumor effect. The other cancer therapeutic agent used for such a purpose may be administered simultaneously with the pharmaceutical composition of the present invention (simultaneously) or at different times (asynchronously or separately). When administered at different times, the other cancer therapeutic agents may be administered consecutively or discontinuously. When administered at different times, the administration interval and order can be changed as appropriate. Such cancer therapeutic agents are not limited as long as they have anti-tumor activity, but examples include at least one selected from the group consisting of endocrine therapy agents, immunotherapeutic agents such as interferon preparations or BCG, and antibodies other than anti-EGFR antibodies or antigen-binding fragments of such antibodies. Here, the antigen-binding fragment refers to a fragment of an antibody that has binding activity to an antigen, and includes Fab, F(ab')2, Fv, scFv, diabody, linear antibody, and multispecific antibody formed from antibody fragments, etc. However, it is not limited to these molecules as long as it has the ability to bind to an antigen.

[0102] The pharmaceutical composition and treatment method of the present invention can also be used in combination with radiation therapy and / or cellular immunotherapy. For example, a cancer patient may receive radiation therapy and / or cellular immunotherapy before and / or after, or simultaneously with, treatment with the pharmaceutical composition of the present invention.

[0103] The pharmaceutical composition and treatment method of the present invention can also be used as adjuvant chemotherapy in combination with surgery. The pharmaceutical composition of the present invention can be administered before surgery to reduce tumor size (neoadjuvant chemotherapy, or neoadjuvant therapy), or after surgery to prevent tumor recurrence (postoperative adjuvant chemotherapy, or adjuvant therapy). The pharmaceutical composition and treatment method of the present invention can also be used as maintenance therapy.

[0104] In addition to the therapeutic uses described above, the pharmaceutical composition and treatment method of the present invention can also be expected to have a preventive effect, such as suppressing the proliferation and even eliminating microscopic metastatic cancer cells. For example, they can be expected to have an effect of suppressing the proliferation and eliminating cancer cells in body fluids during the metastasis process, as well as an effect of suppressing the proliferation and eliminating microscopic cancer cells immediately after metastasis to any tissue. Therefore, they can be expected to have an inhibitory and preventive effect on cancer metastasis, particularly after surgical removal of cancer.

[0105] The pharmaceutical compositions of the present invention can be administered as pharmaceutical compositions containing one or more pharmaceutically compatible ingredients. The substances used in the pharmaceutical compositions of the present invention can be appropriately selected from formulation additives and other substances commonly used in this field, depending on the dosage and administration concentration. For example, the pharmaceutical composition typically contains one or more pharmaceutical carriers (e.g., a sterile liquid). Liquids include, for example, water and oils (petroleum, animal, plant, or synthetic). Oils may include, for example, peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is a more typical carrier when the pharmaceutical composition is administered intravenously. Saline solutions, as well as aqueous dextrose and glycerol solutions, can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients can be appropriately selected from those known in the art. The pharmaceutical composition may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired. Examples of suitable pharmaceutical carriers are described in E. W. The formulation corresponds to the mode of administration.

[0106] Various delivery systems are known and can be used to administer the pharmaceutical compositions of the present invention. Routes of introduction can include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, and subcutaneous routes. Administration can be, for example, by infusion or bolus injection. In certain preferred embodiments, administration of the antibody-drug conjugate is by infusion. Parenteral administration is a preferred route of administration.

[0107] When the anti-EGFR antibody-drug conjugate used in the present invention is administered to a human, the dose and interval of administration are not limited, and for example, about 0.001 to 100 mg / kg can be administered once or multiple times at intervals of once every 1 to 180 days.

[0108] The immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug used in the present invention can be administered at a single dose of 0.1 mg to 6000 mg, and preferably at a single dose of 10 mg to 6000 mg.

[0109] The immune checkpoint inhibitor according to the present invention can be administered to a human at intervals of 1 to 4 times every 1 to 56 days, and preferably at intervals of once every 7 days, once every 14 days, once every 21 days, once every 28 days, or once every 42 days. The chemotherapeutic agent or molecular targeted drug can be administered to a human at intervals of 1 to 3 times per day, or 1 to 4 times per 1 to 56 days, and preferably at intervals of once per day, twice per day, or once every 7 days, once every 14 days, once every 21 days, once every 28 days, or once every 42 days.

[0110] The present invention will be specifically described with reference to the following examples, but the present invention is not limited to these examples. Furthermore, these examples should not be construed as limiting in any sense.

[0111] (Production Example 1) Synthesis of anti-EGFR antibody-drug conjugate 1 (EGFR-ADC (1))

[0112] (Production Example 1-1) Production of anti-EGFR antibody 1 According to the production method described in WO 2020 / 050406, an anti-EGFR antibody (referred to as "anti-EGFR antibody 1" in the present invention) was produced, comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 1 ( FIG. 1 ) and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 2 ( FIG. 2 ).

[0113] (Production Example 1-2) Synthesis of Glycochain Remodeling Antibody 1 Anti-EGFR antibody 1-[SG-(N ３ ) ２ ] ２ Preparation of [Synthetic scheme] (Step 1) Preparation of (Fucα1,6)GlcNAc-anti-EGFR antibody 1 A solution of wild-type EndoS in phosphate buffered saline (0.150 mL, 7.52 mg / mL) was added to a solution of anti-EGFR antibody 1 prepared according to Production Example 1-1 in phosphate buffered saline (20 mL, 11.25 mg / mL, pH 6.0), and the mixture was shaken at 30°C for 18 hours. The progress of the reaction was monitored using an Agilent 2100 Bioanalyzer Electrophoresis System (Agilent Technologies). After completion of the reaction, the mixture was purified by affinity chromatography and hydroxyapatite column chromatography according to the following methods.

[0114] (1) Purification by affinity chromatography Purification apparatus: AKTA avant 25 (GE Healthcare) Column: HiTrap rProtein A FF (5 mL) (GE Healthcare) Flow rate: 5 mL / min (1.25 mL / min during charging) For binding to the column, the reaction solution was added directly to the column, and binding buffer [20 mM phosphate buffer (pH 6.0)] was flowed at 1.25 mL / min for 2 CV, followed by 5 mL / min for 5 CV. For intermediate washing, 15 CV of washing solution [20 mM phosphate buffer (pH 7.0), 0.5 M sodium chloride solution] was flowed. For elution, 6 CV of elution buffer (IgG Elution buffer, Thermo Scientific) was flowed. The eluate was immediately neutralized with 1 M Tris buffer (pH 9.0). The fractions containing the target product were subjected to buffer exchange with 5 mM phosphate buffer, 50 mM 2-morpholinoethanesulfonic acid (MES) solution (pH 6.8) according to the method described in Common Procedure C. The antibody concentration in the resulting buffer was measured according to the method described in Common Procedure B, and a solution of the crudely purified title antibody was obtained.

[0115] (2) Purification by Hydroxyapatite Chromatography Purification apparatus: AKTA Avant 25 Column: Bio-Scale Mini CHT Type I Cartridge (5 mL) (BIO-RAD) Flow rate: 5 mL / min (1.25 mL / min during charging) The solution obtained in (1) above was added to the column, and Solution A [5 mM phosphate buffer, 50 mM MES solution (pH 6.8)] was run at 1.25 mL / min for 2 CV, and then at 5 mL / min for 3 CV. Then, Solution A and Solution B [5 mM phosphate buffer, 50 mM MES solution (pH 6.8), 2 M sodium chloride solution] were used for elution. The elution conditions were Solution A: Solution B = 100:0 to 0:100 (5 CV). Furthermore, 5 CV of a wash solution [500 mM phosphate buffer (pH 6.5)] was applied. The fraction containing the target substance was subjected to buffer exchange with 20 mM phosphate buffer (pH 6.0) according to the method described in Common Procedure C. The antibody concentration in the resulting buffer was measured according to the method described in Common Procedure B, and a 20 mM phosphate buffer solution of the title antibody (17 mL, 12.06 mg / mL, pH 6.0) was obtained.

[0116] (Step 2) Anti-EGFR antibody 1-[SG-(N ３ ) ２ ] ２ The antibody obtained in step 1 above was dissolved in 20 mM phosphate buffer (8.5 mL, 12.06 mg / mL, pH 6.0) and [N ３ -PEG(3)] ２ A solution of 34 mg of SG(10)Ox in 0.340 mL of water and 0.427 mL of EndoS (D233Q / Q303L) in phosphate buffered saline (4.8 mg / mL) were added and the mixture was shaken at 30°C for 4 hours. The progress of the reaction was monitored using an Agilent 2100 Bioanalyzer electrophoresis system. [N ３ -PEG(3)] ２ A solution of -SG(10)Ox (6.8 mg) and EndoS (D233Q / Q303L) in phosphate-buffered saline (0.085 mL, 4.8 mg / mL) was added, and the mixture was shaken at 30°C for 3 hours. After completion of the reaction, purification by affinity chromatography and hydroxyapatite chromatography was carried out as in step 1 above. The fraction containing the target product was subjected to buffer exchange with phosphate-buffered saline (pH 6.0) according to the method described in Common Procedure C. The antibody concentration in the resulting buffer was measured according to the method described in Common Procedure B, and a solution of the title antibody in phosphate-buffered saline (8 mL, 11.09 mg / mL, pH 6.0) was obtained.

[0117] (Production Example 1-3) Synthesis of anti-EGFR antibody-drug conjugate 1 (EGFR-ADC (1)) A solution of glycochain remodeling antibody 1 in phosphate buffered saline (2.00 mL, 11.09 mg / mL, pH 6.0) was diluted with propylene glycol (1.00 mL). To this solution was added drug linker 17a (bis(N,N-diethylethanaminium) N-[4-(11,12-didehydrodibenzo[b,f]azocin-5(6H)-yl)-4-oxobutanoyl]glycylglycyl-L-phenylalanyl-N-[(2-{9-[(5R,7R,8R,12aR,14R,15R,15aR,16R)-15-fluoro-16-hydroxy-2,10-dioxo-2,10-disulfide-14-(6,7,8,9-tetrahydro-2H-2,3,5,6-tetraazabenzo[cd]azulen-2-yl)octahydro-2H,10H,12H-5,8-methano-2λ ５ , 10λ ５ A mixture of a 10 mM dimethyl sulfoxide solution (0.123 mL, 8 equivalents per antibody molecule) of -furo[3,2-l][1,3,6,9,11,2,10]pentaoxadiphosphacyclotetradecin-7-yl]-6-oxo-6,9-dihydro-1H-purin-1-yl}ethoxy)methyl]glycinamide (prepared by the method described in WO 2020 / 050406 )) and propylene glycol (0.877 mL) was added, and the reaction was carried out at room temperature for 43 hours using a tube rotator (MTR-103, AS ONE Corporation). The reaction solution was purified according to the method described in Common Procedure D, yielding an ABS solution (12.0 mL) of the desired antibody-drug conjugate. Analysis was carried out according to the methods described in Common Procedures E and G, and the following results were obtained. Antibody concentration: 1.40 mg / mL Antibody yield: 16.77 mg (76%) Average number of drugs bound: 3.8

[0118] (Production Example 2) Synthesis of anti-EGFR antibody-drug conjugate 2 (EGFR-ADC (2))

[0119] (Production Example 2-1) Preparation of anti-EGFR antibody 1 Anti-EGFR antibody 1 was prepared in the same manner as in Production Example 1-1.

[0120] (Production Example 2-2) Synthesis of Glycochain Remodeling Antibody 2 Anti-EGFR Antibody 1-[MSG1-(N３ )] ２ Preparation of [Synthetic scheme] (Step 1) The antibody obtained in Step 1 of Production Example 1-2 was dissolved in 20 mM phosphate buffer (8.5 mL, 12.06 mg / mL, pH 6.0) and [N ３ A solution of [-PEG(3)]-MSG1(9)Ox (19 mg) in water (0.190 mL) was carried out in the same manner as in Step 2 of Preparation Example 1-2 to give a solution of the title antibody in phosphate buffered saline (8.5 mL, 10.58 mg / mL, pH 6.0).

[0121] (Preparation Example 2-3) Anti-EGFR antibody 1-drug conjugate 2 (synthesis of EGFR-ADC (2)) A phosphate-buffered saline solution of glycosylation remodeling antibody 2 (2.00 mL, 10.58 mg / mL, pH 6.0) and a 10 mM dimethyl sulfoxide solution of drug linker 17a (0.117 mL) were used to carry out the procedures (reaction, purification, and analysis) according to the method described in Preparation Example 1-3, to obtain an ABS solution (12 mL) of the desired antibody-drug conjugate. The analytical results are as follows: Antibody concentration: 1.45 mg / mL Antibody yield: 17.46 mg (82%) Average number of drugs bound: 1.9

[0122] Common Procedure A: Concentration of Aqueous Antibody Solution The antibody or antibody-drug conjugate solution was placed in an Amicon (registered trademark) Ultra centrifugal filter device (50,000 NMWL, Merck Millipore Ltd.), and the antibody or antibody-drug conjugate solution was concentrated by centrifugation (centrifugation at 2000 G to 4000 G for 5 to 20 minutes) using a centrifuge (Allegra X-15R, Beckman Coulter, Inc.).

[0123] Common Procedure B: Measurement of Antibody Concentration Antibody concentrations were measured using a UV measurement device (Nanodrop 1000, Thermo Fisher Scientific, Inc.) according to the method specified by the manufacturer.

[0124] Common Procedure C: Buffer Exchange of Antibody A buffer solution (phosphate buffered saline (pH 6.0), phosphate buffer (pH 6.0), etc.) was added to an aqueous antibody solution, and the solution was concentrated according to the method described in Common Procedure A. After repeating this procedure several times, the antibody concentration was measured according to the method described in Common Procedure B. An appropriate buffer solution (phosphate buffered saline (pH 6.0), phosphate buffer (pH 6.0), etc.) was added to this antibody buffer solution to prepare an antibody buffer solution of the desired concentration (for example, about 10 mg / mL).

[0125] Common Procedure D: Purification of Antibody-Drug Conjugate (Gel Filtration Chromatography) A NAP column (NAP-5, NAP-10, NAP-25 (manufactured by GE Healthcare)) was equilibrated with acetate buffer (10 mM acetate buffer, 5% sorbitol, pH 5.5; referred to herein as ABS) or another appropriate buffer. The antibody-drug conjugate reaction solution was charged onto this NAP column, and a manufacturer-specified amount of buffer was allowed to flow down by gravity, and the antibody fraction was collected. This fraction was again charged onto the NAP column, and a manufacturer-specified amount of buffer was allowed to flow down by gravity, and the antibody fraction was collected. This procedure was repeated two to three times in total to obtain an antibody-drug conjugate from which unbound drug linker, dimethyl sulfoxide, and propylene glycol had been removed. If necessary, the concentration of the antibody-drug conjugate solution was adjusted by common procedures A and C.

[0126] Common Procedure E: Measurement of antibody concentration in antibody-drug conjugate and average number of drugs bound per antibody molecule (UV method) The bound drug concentration in an antibody-drug conjugate can be calculated by measuring the absorbance of an aqueous antibody-drug conjugate solution at two wavelengths, 280 nm and 250 nm, using an absorptiometer (UV / VIS Spectrometer Lambda 25, PerkinElmer, Inc.), and then performing the following calculation. Since the total absorbance at a certain wavelength is equal to the sum of the absorbances of all absorbing chemical species present in the system (additivity of absorbance), assuming that there is no change in the molar extinction coefficients of the antibody and drug before and after conjugation of the antibody with the drug, the antibody concentration and drug concentration in the antibody-drug conjugate are expressed by the following relationship: A２８０ = A Ｄ , ２８０ +A Ａ , ２８０ = ε Ｄ , ２８０ C Ｄ +ε Ａ , ２８０ C Ａ Formula (I) A ２５０ = A Ｄ , ２５０ +A Ａ , ２５０ = ε Ｄ , ２５０ C Ｄ +ε Ａ , ２５０ C Ａ Formula (II) where A ２８０ indicates the absorbance of an aqueous solution of an antibody-drug conjugate at 280 nm, and A ２５０ indicates the absorbance of an aqueous solution of an antibody-drug conjugate at 250 nm, and A Ａ , ２８０ indicates the absorbance of the antibody at 280 nm, and A Ａ , ２５０ indicates the absorbance of the antibody at 250 nm, and A Ｄ , ２８０ denotes the absorbance of the conjugate precursor at 280 nm, and A Ｄ , ２５０ denotes the absorbance of the conjugate precursor at 250 nm, and ε Ａ , ２８０ denotes the molar extinction coefficient of the antibody at 280 nm, and ε Ａ , ２５０ denotes the molar extinction coefficient of the antibody at 250 nm, and ε Ｄ , 280 denotes the molar extinction coefficient of the conjugate precursor at 280 nm, and ε Ｄ , ２５０ denotes the molar extinction coefficient of the conjugate precursor at 250 nm, and C Ａ indicates the antibody concentration in the antibody-drug conjugate, and C Ｄ denotes the drug concentration in the antibody-drug conjugate, where ε Ａ , ２８０ , ε Ａ , ２５０ , ε Ｄ ,２８０ , ε Ｄ , ２５０ A value prepared in advance (calculated estimated value or actual measured value) is used for 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). Ａ , ２５０ is the difference between the measured value obtained from UV measurement of the antibody and ε Ａ , ２８０ In the examples, the molar extinction coefficient of anti-EGFR antibody 1 was calculated as ε Ａ , ２８０ = 203460 and ε Ａ , ２５０ = 63051 or 63370 was used. Ｄ , ２８０ and ε Ｄ , ２５０ The A of the antibody-drug conjugate solution was measured using the Beer-Lambert law (absorbance = molar concentration × molar extinction coefficient × cell path length) by dissolving the conjugate precursor to be used at a certain molar concentration. The A of the antibody-drug conjugate solution was measured using the Beer-Lambert law (absorbance = molar concentration × molar extinction coefficient × cell path length). The molar extinction coefficient of the conjugate precursor in the examples was obtained by UV measurement. ２８０ and A ２５０ These values ​​are substituted into the equations (I) and (II) to solve the simultaneous equations, thereby obtaining C Ａ and C Ｄ Furthermore, C Ｄ C Ａ By dividing by this, the average number of drugs bound per antibody molecule can be calculated.

[0127] Common Procedure F: Measurement of antibody concentration in antibody-drug conjugates and average number of drugs bound per antibody molecule (reverse-phase high-performance liquid chromatography: RP-HPLC) The antibody concentration in antibody-drug conjugates and average number of drugs bound per antibody molecule can be determined by high-performance liquid chromatography analysis using the following method, in addition to the above-mentioned common procedure E.

[0128] [F-1. Preparation of sample for HPLC analysis (reduction of antibody-drug conjugate)] The antibody-drug conjugate solution (approximately 1 mg / mL, 60 μL) was mixed with an aqueous dithiothreitol (DTT) solution (100 mM, 15 μL). The mixture was incubated at 37°C for 30 minutes to cleave the disulfide bond between the L chain and H chain of the antibody-drug conjugate. This reaction solution was used directly for HPLC analysis.

[0129] [F-2. HPLC analysis] Typical analytical conditions are as follows: HPLC system: Agilent 1290 HPLC system (Agilent Technologies) Detector: UV spectrophotometer (measurement wavelength: 280 nm) Column: Acquity BEH Phenyl (2.1 x 50 mm, 1.7 μm, manufactured by Waters) Column temperature: 75°C Flow rate: 0.8 mL / min Sample injection volume: 10 μL Mobile phase A: 0.1% trifluoroacetic acid (TFA), 15% isopropyl alcohol aqueous solution Mobile phase B: 0.075% TFA, 15% isopropyl alcohol acetonitrile solution Gradient program (mobile phase B): 14%-36% (0 min-15 min), 36%-80% (15-17 min), 80%-14% (17 min-17.1 min), 14%-14% (17.1 min-23 min)

[0130] [F-3. Data Analysis] [F-3-1] In the case of sugar chain conjugation in the SPAAC reaction, the H chains to which drugs are bound (H chains to which one drug is bound: H1, H chains to which two drugs are bound: H2) become more hydrophobic and have longer retention times in proportion to the number of drugs bound, compared to the L chain (L0) and H chain (H0) of an antibody to which no drug is bound. Therefore, they are eluted in the order of L0, H0, H1, and H2 in principle. By comparing the retention times of L0 and H0, the detected peak can be assigned to either L0, H0, H1, or H2. Similarly, in the case of cysteine ​​conjugation, the hydrophobicity of drug-bound L chains (L chains with one drug bound: L1) and drug-bound H chains (H chains with one drug bound: H1, H chains with two drugs bound: H2, H chains with three drugs bound: H3) increases in proportion to the number of drugs bound, and the retention time increases, so they are eluted in the order of L0, L1, H0, H1, H2, and H3 in principle. By comparing the retention times of L0 and H0, the detected peak can be assigned to either L0, L1, H0, H1, H2, or H3.

[0131] [F-3-2] Because the drug linker has UV absorption, in the case of sugar chain conjugation in the SPAAC reaction, the peak area was corrected according to the number of drug linkers bound using the molar extinction coefficients of the H chain and drug linker according to the following formula: In the case of cysteine ​​conjugation in which the drug is also bound to the L chain, the peak area was similarly corrected for the L chain.

[0132]

[0133] Here, the molar extinction coefficients (280 nm) of the L chain and H chain of each antibody were estimated values ​​calculated by the known calculation method described in Common Procedure E. In the case of anti-EGFR antibody 1, 23232 was used as the molar extinction coefficient of the L chain, and 78498 was used as the molar extinction coefficient of the H chain. For the molar extinction coefficient (280 nm) of the drug linker, in the case of sugar chain conjugation by SPAAC reaction, the actual measured value of the conjugation precursor was used, and in the case of cysteine ​​conjugation, the actual measured value of the compound in which the conjugation precursor was reacted with mercaptoethanol or N-acetylcysteine ​​to convert the maleimide group to succinimide thioether was used.

[0134] [F-3-3] The ratio (%) of each chain peak area to the total corrected peak area was calculated according to the following formula.

[0135]

[0136] [F-3-4] The average number of drugs bound per antibody molecule (DAR) in the antibody-drug conjugate was calculated according to the following formula.

[0137]

[0138] [F-3-5] The antibody concentration in the antibody-drug conjugate was calculated according to the following formula.

[0139]

[0140] Here, the absorbance (280 nm) of the antibody-drug conjugate was determined using the actual measured value of the antibody-drug conjugate aqueous solution. The dilution factor indicates how many times the antibody-drug conjugate aqueous solution was diluted when measuring absorbance, and is usually 4-fold. The molar extinction coefficient (280 nm) of the antibody was an estimated value calculated using the known calculation method described in Common Procedure E. The average number of drugs bound was used as the value obtained in [F-3-4]. For the molar extinction coefficient (280 nm) of the drug linker, the actual measured value of the conjugation precursor was used in the case of sugar chain conjugation by SPAAC reaction, and for cysteine ​​conjugation, the actual measured value of the compound obtained by reacting the conjugation precursor with mercaptoethanol or N-acetylcysteine ​​to convert the maleimide group to succinimide thioether was used.

[0141] Common Procedure G: Measurement of antibody concentration in antibody-drug conjugates and average number of drugs bound per antibody molecule (hydrophobic interaction-high performance liquid chromatography: HI-HPLC) The antibody concentration in antibody-drug conjugates and average number of drugs bound per antibody molecule can be determined by high performance liquid chromatography analysis using the following method, in addition to the above-mentioned common procedures E and F.

[0142] [G-1. Preparation of sample for HPLC analysis] The antibody-drug conjugate solution (approximately 1 mg / mL, 60 μL) was used directly for HPLC analysis.

[0143] [G-2. HPLC Analysis] The following two typical analytical conditions are used. HPLC system: SHIMADZU CBM-20A (Shimadzu Corporation) Detector: UV spectrophotometer (measurement wavelength: 280 nm) Column: TSK-gel Butyl-NPR (4.6 x 100 mm, 2.5 μm, manufactured by TOSOH) Column temperature: constant temperature around 25°C Mobile phase A: 25 mM phosphate buffer (pH = 7.0) containing 1.5 M ammonium sulfate Mobile phase B: 25 mM phosphate buffer (pH = 7.0) / isopropyl alcohol mixture (3:1) Flow rate: 0.8 mL / min Sample injection volume: 15 μL Gradient program (mobile phase B): 10% - 15% (0 min - 5 min), 15% - 65% (5 min - 20 min) or HPLC system: SHIMADZU CBM-20A (Shimadzu Corporation) Detector: UV spectrophotometer (measurement wavelength: 280 nm) Column: PolyPROPYL A (4.6 x 100 mm, 3 μm, 1500 Å, manufactured by PolyLC) Column temperature: constant temperature around 40°C Mobile phase A: 20 mM phosphate buffer (pH = 7.4) containing 1.5 M ammonium sulfate Mobile phase B: 20 ​​mM phosphate buffer (pH = 7.4) Flow rate: 0.8 mL / min Sample injection volume: 15 μL Gradient program (mobile phase B): 40% - 80% (0 min - 20 min)

[0144] [G-3. Data Analysis] [G-3-1] Since hydrophobicity increases in proportion to the number of drugs bound to the antibody and retention time increases, in the case of sugar chain conjugation in the SPAAC reaction, elution generally occurs in the order of DAR=0, DAR=2, and DAR=4. By comparing the retention time with DAR=0, the detected peak can be assigned to either DAR=2 or DAR=4. Depending on the type of antibody and drug linker, peaks with DAR=1 and DAR=3 may also be detected. The DAR of the detected peak may also be estimated by measuring the mass spectrum after fractionating the peak by HI-HPLC.

[0145] [G-3-2] Because the drug linker has UV absorption, the peak area value was corrected according to the number of drug linkers bound using the molar absorption coefficients of the antibody and drug linker according to the following formula.

[0146]

[0147] Here, the molar extinction coefficient (280 nm) of the antibody was an estimated value calculated by the known calculation method described in Common Procedure E. The molar extinction coefficient (280 nm) of the drug linker was the actually measured value of the conjugation precursor.

[0148] [G-3-3] The antibody peak area ratio (%) to the total corrected peak area was calculated according to the following formula.

[0149]

[0150] [G-3-4] The average number of drugs bound per antibody molecule in the antibody-drug conjugate was calculated according to the following formula.

[0151]

[0152] [G-3-5] The antibody concentration in the antibody-drug conjugate was calculated according to the formula described in [F-3-5], and the average number of bound drugs was calculated using the value obtained in [G-3-4].

[0153] The EGFR-ADC (1) and EGFR-ADC (2) obtained in Production Examples 1 and 2 have the following structures. In the above formula, m ２The anti-EGFR antibody-drug conjugate of m = 2 is "EGFR-ADC (1)" ２ The anti-EGFR antibody-drug conjugate of EGFR-ADC (2) is designated as EGFR-ADC (2).

[0154] (Antitumor test) Measurement and calculation formula: In all studies, the major and minor axes of the tumor were measured two to four times a week using an electronic digital caliper (CD-15CX, Mitutoyo Corp.), and the tumor volume (mm ３ The calculation formula is as follows: Tumor volume (mm ３ ) = 1 / 2 x major axis (mm) x [minor axis (mm)] ２

[0155] In the following test examples, the tumor volume was 50 mm ３ The following individuals are referred to as "CR individuals".

[0156] (Test Example 1) Antitumor Test (1) EMT6-chimeraEGFR cells were prepared by introducing a human-mouse chimeric EGFR gene, in which the epitope site of an anti-human EGFR antibody had been humanized, into the mouse colon cancer cell line EMT6 purchased from American Type Culture Collection. EMT6-chimeraEGFR cells were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and seven days later, the mice were randomly assigned to groups. EGFR-ADC (1) was administered once into the tail vein at a dose of 0.3 mg / kg on Day 7. Anti-PD-1 surrogate antibody (Anti-mPD-1 mIgG1e3, Invivogen) was administered at a dose of 5 mg / kg into the tail vein a total of four times on Day 7, Day 11, Day 14, and Day 18. Each group had six mice. The results are shown in Figure 4. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the anti-PD-1 surrogate antibody administration group, and the black inverted triangle line indicates the group administered with a combination of EGFR-ADC(1) and anti-PD-1 surrogate antibody. The vertical axis represents tumor volume (mm ３), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. Tumor growth was suppressed in the EGFR-ADC (1)-administered group and the anti-PD-1 surrogate antibody-administered group, but 0 out of 6 mice achieved CR as of Day 25. In contrast, tumor growth was significantly suppressed in the group administered a combination of EGFR-ADC (1) and anti-PD-1 surrogate antibody, with 5 out of 6 mice achieving CR as of Day 25. From the above, the strong anti-tumor combined effect of EGFR-ADC (1) and anti-PD-1 surrogate antibody was confirmed.

[0157] (Test Example 2) Antitumor Test (2) CT26.WT-chimeraEGFR cells were prepared by introducing a human-mouse chimeric EGFR gene, in which the epitope site of the anti-human EGFR antibody had been humanized, into the mouse colon cancer cell line CT26.WT purchased from American Type Culture Collection. CT26.WT-chimeraEGFR cells were subcutaneously implanted into the right axilla of BALB / c mice (Day 0), and 7 days later, the mice were randomly assigned to groups. EGFR-ADC (1) was administered once into the tail vein at a dose of 0.1 mg / kg on Day 7. Anti-PD-1 surrogate antibody (Anti-mPD-1 mIgG1e3, Invivogen) was administered at a dose of 5 mg / kg into the tail vein a total of three times on Day 7, Day 11, and Day 14. Each group consisted of six mice. The results are shown in Figure 5. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the anti-PD-1 surrogate antibody administration group, and the black inverted triangle line indicates the group administered with a combination of EGFR-ADC(1) and anti-PD-1 surrogate antibody. The vertical axis represents tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 18, compared to the vehicle-administered group, the tumor volume was 38% smaller in the EGFR-ADC(1)-administered group and 31% smaller in the anti-PD-1 surrogate antibody-administered group. In contrast, tumor growth was significantly suppressed in the group administered a combination of EGFR-ADC(1) and anti-PD-1 surrogate antibody, and the tumor volume at day 18 was 82% smaller than that of the vehicle-administered group. From the above, the strong anti-tumor combined effect of EGFR-ADC(1) and anti-PD-1 surrogate antibody was confirmed.

[0158] (Test Example 3) Antitumor Test (3) Test Examples 3 and 4 were conducted simultaneously, and therefore Figures 6 and 7 were prepared using the same data for the vehicle-administered group and the EGFR-ADC (1)-administered group. CT26.WT-chimeraEGFR cells prepared in Test Example 2 were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and seven days later, mice were randomly assigned to groups. EGFR-ADC (1) was administered intravenously at a dose of 0.3 mg / kg once on Day 7. The anti-PD-L1 surrogate antibody (clone number 10F.9G2) was administered intravenously at a dose of 10 mg / kg once on Day 7. Each group contained eight mice. The results are shown in Figure 6. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the anti-PD-L1 surrogate antibody administration group, and the black inverted triangle line indicates the EGFR-ADC(1) and anti-PD-L1 surrogate antibody combined administration group. The vertical axis represents tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At Day 17, tumor volume was 63% smaller in the EGFR-ADC(1)-administered group and 12% smaller in the anti-PD-L1 surrogate antibody-administered group compared to the vehicle-administered group. Furthermore, 0 out of 6 mice achieved CR at Day 17. In contrast, tumor growth was significantly suppressed in the group administered a combination of EGFR-ADC(1) and anti-PD-L1 surrogate antibody, and tumor volume at Day 17 was 80% smaller than in the vehicle-administered group. Furthermore, 2 out of 6 mice achieved CR at Day 17. These results confirmed the strong anti-tumor effect of the combination of EGFR-ADC(1) and anti-PD-L1 surrogate antibody.

[0159] (Test Example 4) Antitumor Test (4) CT26.WT-chimeraEGFR cells prepared in Test Example 2 were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and seven days later, the mice were randomly assigned to groups. EGFR-ADC (1) was administered intravenously into the tail vein at a dose of 0.3 mg / kg once on Day 7. Anti-CTLA4 surrogate antibody (clone number 9D9) was administered intravenously into the tail vein at a dose of 10 mg / kg twice on Day 7 and Day 14. Each group consisted of eight mice. The results are shown in Figure 7. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the anti-CTLA4 surrogate antibody administration group, and the black inverted triangle line indicates the EGFR-ADC(1) and anti-CTLA4 surrogate antibody combined administration group. The vertical axis indicates tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 17, compared to the vehicle-administered group, the tumor volume was 63% smaller in the EGFR-ADC(1)-administered group and 21% smaller in the anti-CTLA4 surrogate antibody-administered group. In contrast, tumor growth was significantly suppressed in the group administered a combination of EGFR-ADC(1) and anti-CTLA4 surrogate antibody, and the tumor volume at day 17 was 86% smaller than that of the vehicle-administered group. From the above, a strong anti-tumor combined effect of EGFR-ADC(1) and anti-CTLA4 surrogate antibody was confirmed.

[0160] (Test Example 5) Antitumor Test (5) Test Example 5 and Test Example 6 were conducted simultaneously, and therefore Figures 8 and 9 were created using the same data for the vehicle-administered group and the EGFR-ADC (2)-administered group. 4T1-chimeraEGFR cells were prepared by introducing a human-mouse chimeric EGFR gene, in which the epitope site of the anti-human EGFR antibody had been humanized, into the mouse breast cancer cell line 4T1 purchased from American Type Culture Collection. 4T1-chimeraEGFR cells were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and 6 days later, the mice were randomly assigned to groups. EGFR-ADC (2) was administered once into the tail vein at a dose of 0.6 mg / kg on Day 7. Cisplatin was administered once into the tail vein at a dose of 5 mg / kg on Day 6. Each group consisted of six mice. The results are shown in Figure 8. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(2) administration group, the black triangle line indicates the cisplatin administration group, and the black inverted triangle line indicates the EGFR-ADC(2) and cisplatin combined administration group. The vertical axis indicates tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 23, tumor volume was 32% smaller in the EGFR-ADC(2)-administered group and 17% smaller in the cisplatin-administered group compared to the vehicle-administered group. In contrast, tumor growth was significantly suppressed in the EGFR-ADC(2) and cisplatin-administered group, and tumor volume at day 23 was 52% smaller than that of the vehicle-administered group. From the above, the strong antitumor effect of the combined use of EGFR-ADC(2) and cisplatin was confirmed.

[0161] (Test Example 6) Antitumor Test (6) The 4T1-chimeraEGFR cells prepared in Test Example 5 were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and six days later, the mice were randomly divided into groups. EGFR-ADC (2) was administered into the tail vein at a dose of 0.6 mg / kg once in total on Day 7. Oxaliplatin was administered into the tail vein at a dose of 5 mg / kg twice in total on Day 6 and Day 13. Each group consisted of six mice. The results are shown in Figure 9. In the figure, the black circle line indicates the vehicle-administered group, the black square line indicates the EGFR-ADC (2)-administered group, the black triangle line indicates the oxaliplatin-administered group, and the black inverted triangle line indicates the group administered with a combination of EGFR-ADC (2) and oxaliplatin. The vertical axis indicates tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 23, tumor volume was 32% smaller in the EGFR-ADC(2)-administered group and 18% smaller in the oxaliplatin-administered group compared to the vehicle-administered group. In contrast, tumor growth was significantly suppressed in the EGFR-ADC(2) and oxaliplatin-administered group, and tumor volume at day 23 was 48% smaller than that of the vehicle-administered group. From the above, the strong antitumor combined effect of EGFR-ADC(2) and oxaliplatin was confirmed.

[0162] (Test Example 7) Antitumor Test (7) Test Examples 7 and 8 were conducted simultaneously, and therefore Figures 10 and 11 were prepared using the same data for the vehicle-administered group and the EGFR-ADC (1)-administered group. CT26.WT-chimeraEGFR cells prepared in Test Example 2 were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and six days later, the mice were randomly assigned to groups. EGFR-ADC (1) was administered intravenously at a dose of 0.25 mg / kg once on Day 7. Cisplatin was administered intravenously at a dose of 3 mg / kg once on Day 6. Each group contained six mice. The results are shown in Figure 10. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the cisplatin administration group, and the black inverted triangle line indicates the EGFR-ADC(1) and cisplatin combined administration group. The vertical axis represents tumor volume (mm ３), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 17, tumor volume was 80% smaller in the EGFR-ADC(1)-administered group and 15% smaller in the cisplatin-administered group compared to the vehicle-administered group. In contrast, tumor growth was significantly suppressed in the EGFR-ADC(1) and cisplatin-administered group, and tumor volume at day 17 was 86% smaller than that of the vehicle-administered group. From the above, a strong antitumor combined effect of EGFR-ADC(1) and cisplatin was confirmed.

[0163] (Test Example 8) Antitumor Test (8) CT26.WT-chimeraEGFR cells prepared in Test Example 2 were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and 6 days later, the mice were randomly divided into groups. EGFR-ADC(1) was administered into the tail vein at a dose of 0.25 mg / kg once on Day 7. Oxaliplatin was administered into the tail vein at a dose of 3 mg / kg twice on Day 6 and Day 13. Each group consisted of 6 mice. The results are shown in Figure 11. In the figure, the black circle line indicates the vehicle-administered group, the black square line indicates the EGFR-ADC(1)-administered group, the black triangle line indicates the oxaliplatin-administered group, and the black inverted triangle line indicates the group administered with a combination of EGFR-ADC(1) and oxaliplatin. The vertical axis indicates tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 17, tumor volume was 80% smaller in the EGFR-ADC(1)-administered group and 47% smaller in the oxaliplatin-administered group compared to the vehicle-administered group. In contrast, tumor growth was significantly suppressed in the EGFR-ADC(1) and oxaliplatin-administered group, and tumor volume at day 17 was 88% smaller than that of the vehicle-administered group. From the above, the strong antitumor effect of the combined use of EGFR-ADC(1) and oxaliplatin was confirmed.

[0164] (Test Example 9) Antitumor Test (9) Test Examples 9 and 10 were conducted simultaneously, and therefore Figures 12 and 13 were prepared using the same data for the vehicle-administered group and the EGFR-ADC (2)-administered group. 4T1-chimeraEGFR cells prepared in Test Example 5 were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and six days later, the mice were randomly assigned to groups. EGFR-ADC (2) was administered intravenously at a dose of 0.6 mg / kg once on Day 7. Gemcitabine was administered intravenously at a dose of 50 mg / kg twice on Days 6 and 13. Each group contained six mice. The results are shown in Figure 12. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(2) administration group, the black triangle line indicates the gemcitabine administration group, and the black inverted triangle line indicates the EGFR-ADC(2) and gemcitabine combined administration group. The vertical axis represents tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 23, tumor volume was 24% smaller in the EGFR-ADC(2)-administered group and 52% smaller in the gemcitabine-administered group compared to the vehicle-administered group. In contrast, tumor growth was significantly suppressed in the EGFR-ADC(2) and gemcitabine-administered group, and tumor volume at day 23 was 65% smaller than that of the vehicle-administered group. From the above, the strong antitumor effect of the combined use of EGFR-ADC(2) and gemcitabine was confirmed.

[0165] (Test Example 10) Antitumor Test (10) The 4T1-chimeraEGFR cells prepared in Test Example 5 were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and six days later, the mice were randomly divided into groups. EGFR-ADC (2) was administered into the tail vein at a dose of 0.6 mg / kg once on Day 7. Fluorouracil was administered into the tail vein at a dose of 50 mg / kg twice on Day 6 and Day 13. Each group consisted of six mice. The results are shown in Figure 13. In the figure, the black circle line indicates the vehicle-administered group, the black square line indicates the EGFR-ADC (2)-administered group, the black triangle line indicates the fluorouracil-administered group, and the black inverted triangle line indicates the group administered with a combination of EGFR-ADC (2) and fluorouracil. The vertical axis indicates tumor volume (mm ３), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 23, tumor volume was 24% smaller in the EGFR-ADC(2)-administered group and 16% smaller in the fluorouracil-administered group compared to the vehicle-administered group. In contrast, tumor growth was significantly suppressed in the EGFR-ADC(2) and fluorouracil-administered group, and tumor volume at day 23 was 39% smaller than that of the vehicle-administered group. From the above, the strong antitumor effect of the combined use of EGFR-ADC(2) and fluorouracil was confirmed.

[0166] (Test Example 11) Antitumor Test (11) Human lung adenocarcinoma cell line NCI-H1975 purchased from American Type Culture Collection was subcutaneously transplanted into the right axilla of BALB / c-nu mice (Day 0), and seven days later, the mice were randomly assigned to groups. EGFR-ADC (1) was administered into the tail vein at a dose of 0.5 mg / kg once on Day 7. Docetaxel was administered into the tail vein at a dose of 15 mg / kg once on Day 7. Each group consisted of six mice. The results are shown in Figure 14. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the docetaxel administration group, and the black inverted triangle line indicates the EGFR-ADC(1) and docetaxel co-administration group. The vertical axis represents tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. Tumor growth was suppressed in the EGFR-ADC(1)-administered group and the docetaxel-administered group, but only two out of six mice achieved CR as of Day 35. In contrast, tumor growth was significantly suppressed in the EGFR-ADC(1) and docetaxel-administered group, and six out of six mice achieved CR as of Day 35. From the above, a strong antitumor combined effect of EGFR-ADC(1) and docetaxel was confirmed.

[0167] (Test Example 12) Antitumor Test (12) An anti-VEGF surrogate antibody was prepared by converting the Fc region of an anti-VEGF antibody (clone number G6-31) to human IgG1 and adding LALA mutations (L234A, L235A). EMT6-chimeraEGFR cells prepared in Test Example 1 were subcutaneously transplanted into the right axilla of BALB / c mice (Day 0), and seven days later, the mice were randomly assigned to groups. EGFR-ADC (1) was administered intravenously at a dose of 0.3 mg / kg once on Day 7. The anti-VEGF surrogate antibody was administered intravenously at a dose of 7.5 mg / kg twice on Day 7 and Day 10. Each group consisted of six mice. The results are shown in Figure 15. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the anti-VEGF surrogate antibody administration group, and the black inverted triangle line indicates the EGFR-ADC(1) and anti-VEGF surrogate antibody combined administration group. The vertical axis indicates tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At day 27, tumor volume was 54% smaller in the EGFR-ADC(1)-administered group and 34% smaller in the anti-VEGF surrogate antibody-administered group compared to the vehicle-administered group. In contrast, tumor growth was significantly suppressed in the group administered a combination of EGFR-ADC(1) and anti-VEGF surrogate antibody, and tumor volume at day 27 was 66% smaller than that of the vehicle-administered group. From the above, a strong anti-tumor combined effect of EGFR-ADC(1) and anti-VEGF surrogate antibody was confirmed.

[0168] (Test Example 13) Antitumor Test (13) Human lung cancer cell line PC-9 purchased from RIKEN BioResource Research Center was subcutaneously transplanted into the right axilla of BALB / c-nu mice (Day 0), and seven days later, the mice were randomly assigned to groups. EGFR-ADC (1) was administered intravenously into the tail vein at a dose of 0.5 mg / kg once on Day 8. Osimertinib mesylate was orally administered at a dose of 25 mg / kg five times daily from Day 7 to Day 11. Each group contained six mice. The results are shown in Figure 16. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the osimertinib mesylate administration group, and the black inverted triangle line indicates the EGFR-ADC(1) and osimertinib mesylate combination administration group. The vertical axis indicates tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. Tumor growth was suppressed in the EGFR-ADC (1)-administered group and the osimertinib mesylate-administered group, but as of Day 28, 0 out of 6 mice had a CR. In contrast, tumor growth was significantly suppressed in the EGFR-ADC (1) and osimertinib mesylate-administered group, and as of Day 28, 6 out of 6 mice had a CR. From the above, the strong antitumor combined effect of EGFR-ADC (1) and osimertinib mesylate was confirmed.

[0169] (Test Example 14) Antitumor Test (14) LL / 2-chimeraEGFR cells were prepared by introducing a human-mouse chimeric EGFR gene, in which the epitope site of the anti-human EGFR antibody had been humanized, into the mouse lung cancer cell line LL / 2 purchased from American Type Culture Collection. LL / 2-chimeraEGFR cells were subcutaneously implanted into the right axilla of C57BL / 6J mice (Day 0), and 6 days later, the mice were randomly assigned to groups. EGFR-ADC (2) was administered at a dose of 0.6 mg / kg into the tail vein once on Day 7. Anti-PD-1 surrogate antibody (Anti-mPD-1 mIgG1e3, Invivogen) was administered into the tail vein at a dose of 5 mg / kg four times on Day 7, Day 10, Day 13, and Day 16. Cisplatin was administered into the tail vein at a dose of 5 mg / kg twice on Day 6 and Day 11. There were six mice in each group. The results are shown in Figures 17 and 18. In the figure, the black circle line represents the vehicle administration group, the black square line represents the EGFR-ADC (2) administration group, the black triangle line represents the anti-PD-1 surrogate antibody administration group, the black inverted triangle line represents the cisplatin administration group, the black diamond line represents the EGFR-ADC (2) and anti-PD-1 surrogate antibody combination administration group, the white circle line represents the EGFR-ADC (2) and cisplatin combination administration group, the white square line represents the anti-PD-1 surrogate antibody and cisplatin combination administration group, and the white triangle line represents the EGFR-ADC (2), anti-PD-1 surrogate antibody and cisplatin combination administration group. The vertical axis represents tumor volume (mm3), and the horizontal axis represents the number of days after tumor transplantation. Tumor growth progressed in the vehicle administration group. At day 20, compared to the vehicle-administered group, the tumor volume was 60% smaller in the EGFR-ADC (2) administration group, 12% smaller in the anti-PD-1 surrogate antibody administration group, and 53% smaller in the cisplatin administration group. Also, at day 20, compared to the vehicle-administered group, the tumor volume in the EGFR-ADC (2) and anti-PD-1 surrogate antibody combination administration group was 54% smaller, and the tumor volume in the anti-PD-1 surrogate antibody and cisplatin combination administration group was 52% smaller. In contrast, tumor growth was significantly suppressed in the EGFR-ADC (2) and cisplatin combination administration group, and the tumor volume at day 20 was 80% smaller than the vehicle-administered group.Furthermore, tumor growth was significantly suppressed in the group administered a combination of EGFR-ADC (2), an anti-PD-1 surrogate antibody, and cisplatin, and the tumor volume at Day 20 was 94% smaller than that of the vehicle-administered group. From the above, the strong antitumor combined effect of EGFR-ADC (2) and cisplatin, and the strong antitumor combined effect of EGFR-ADC (2), an anti-PD-1 surrogate antibody, and cisplatin were confirmed.

[0170] (Test Example 15) Antitumor Test (15) Human lung adenocarcinoma cell line NCI-H1975 purchased from American Type Culture Collection was subcutaneously transplanted into the right axilla of BALB / c-nu mice (Day 0), and 13 days later, mice were randomly assigned to groups. EGFR-ADC (1) was administered intravenously into the tail vein at a dose of 0.3 mg / kg once on Day 13. Irinotecan hydrochloride hydrate was administered intraperitoneally at a dose of 40 mg / kg four times on Days 13, 20, 27, and 34. Each group contained six mice. The results are shown in Figure 19. In the figure, the black circle line indicates the vehicle administration group, the black square line indicates the EGFR-ADC(1) administration group, the black triangle line indicates the irinotecan hydrochloride hydrate administration group, and the black inverted triangle line indicates the EGFR-ADC(1) and irinotecan hydrochloride hydrate combination administration group. The vertical axis represents tumor volume (mm ３ ), the horizontal axis indicates the number of days after tumor implantation. Tumor growth progressed in the vehicle-administered group. At Day 38, tumor volume was 76% smaller in the EGFR-ADC(1)-administered group and 57% smaller in the irinotecan hydrochloride hydrate-administered group compared to the vehicle-administered group. In contrast, tumor growth was significantly suppressed in the group administered a combination of EGFR-ADC(1) and irinotecan hydrochloride hydrate, and tumor volume at Day 38 was 92% smaller than that of the vehicle-administered group. From the above, a strong antitumor combined effect of EGFR-ADC(1) and irinotecan hydrochloride hydrate was confirmed.

[0171] In Test Examples 1 to 15, no particularly noticeable findings such as weight loss were observed in any of the single agent and combination administration groups.

[0172] SEQ ID NO: 1: Amino acid sequence of the heavy chain of anti-EGFR antibody 1 SEQ ID NO: 2: Amino acid sequence of the light chain of anti-EGFR antibody 1 SEQ ID NO: 3: Amino acid sequence of CDRH1 of anti-EGFR antibody 1 SEQ ID NO: 4: Amino acid sequence of CDRH2 of anti-EGFR antibody 1 SEQ ID NO: 5: Amino acid sequence of CDRH3 of anti-EGFR antibody 1 SEQ ID NO: 6: Amino acid sequence of CDRL1 of anti-EGFR antibody 1 SEQ ID NO: 7: Amino acid sequence of CDRL3 of anti-EGFR antibody 1

Claims

1. a) A pharmaceutical composition comprising an anti-EGFR antibody-drug conjugate, wherein a) the anti-EGFR antibody-drug conjugate and b) one or more selected from an immune checkpoint inhibitor, a chemotherapeutic agent, and a molecular targeted drug are used in combination, and the anti-EGFR antibody-drug conjugate is represented by the following formula (I): (In the formula, m ２ represents an integer of 1 or 2, L represents a linker connecting the N297 sugar chain and D, -Lb-La-Lp-Lc-* (wherein the asterisk indicates binding to drug D, and Lb is: (In the structural formula of Lb shown above, the asterisk indicates bonding to La, and the wavy line indicates bonding to the N297 sugar chain), where La is -C(=O)-CH ２ CH ２ Lp represents -GGFG- or -GGPI-, Lc represents -NH-CH ２ -), Ab is an anti-EGFR antibody or an antigen-binding fragment thereof comprising a heavy chain comprising CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and a light chain comprising CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, CDRL2 consisting of the amino acid sequence represented by DAS, and CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 7, wherein the N297 sugar chain is N297-(Fuc)MSG1 having the structure represented by the following formula: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of the 1-3 chain side of the branched chain of β-Man in the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1- or 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), or N297-(Fuc)SG: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), and D represents (wherein the asterisk indicates bonding to L, and W represents -NH-).

2. D is The pharmaceutical composition according to claim 1, which is represented by the formula: (wherein the asterisk indicates binding to L).

3. The anti-EGFR antibody-drug conjugate has the formula: (Wherein Ab, N297 sugar chain and m ２ The pharmaceutical composition according to claim 1 or 2, wherein 4. The anti-EGFR antibody-drug conjugate has the formula: In the formula, m ２ is 2, the Ab comprises a heavy chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and a light chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2, and the N297 sugar chain is N297-(Fuc)SG having a structure represented by the following formula: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, the asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring, and n ５ The pharmaceutical composition according to any one of claims 1 to 3, wherein R is an integer of 1 to 3.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the Ab comprises a heavy chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 119 of SEQ ID NO: 1, and a light chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 107 of SEQ ID NO:

2.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the Ab comprises a heavy chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence shown in SEQ ID NO: 1, and a light chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence shown in SEQ ID NO:

2.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the Ab comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 1 and a light chain consisting of the amino acid sequence shown in SEQ ID NO:

2.

8. The pharmaceutical composition of any one of claims 1 to 5, wherein the Ab comprises an antibody or an antigen-binding fragment of said antibody contained in the pharmaceutical composition of any one of claims 1 to 7, and comprises: (i) a heavy chain consisting of an amino acid sequence in which one or two amino acids are deleted from the carboxyl terminus of the amino acid sequence of the heavy chain; and (ii) a light chain.

9. The pharmaceutical composition according to any one of claims 1 to 8, wherein a) an anti-EGFR antibody-drug conjugate and b) an immune checkpoint inhibitor are used in combination, and the immune checkpoint inhibitor is an antibody selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-TIGIT antibody, and an anti-LAG-3 antibody, an antigen-binding fragment thereof, or a compound comprising any of them.

10. The pharmaceutical composition according to claim 9, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, cemiplimab, or dostarlimab.

11. The pharmaceutical composition according to claim 9, wherein the anti-PD-L1 antibody is atezolizumab, duvalumab, or avelumab.

12. The pharmaceutical composition of claim 9, wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab.

13. The pharmaceutical composition according to claim 9, wherein the anti-TIGIT antibody is tiragolumab or vibostolimab.

14. The pharmaceutical composition of claim 9, wherein the anti-LAG-3 antibody is leratolimab.

15. The pharmaceutical composition according to any one of claims 1 to 8, wherein a) the anti-EGFR antibody-drug conjugate and b) a chemotherapeutic agent are used in combination, and the chemotherapeutic agent is an antimetabolite, a tubulin inhibitor, an alkylating agent, a platinum compound, or a topoisomerase inhibitor.

16. The pharmaceutical composition of claim 15, wherein the antimetabolite is gemcitabine or a pharmaceutically acceptable salt thereof.

17. The pharmaceutical composition of claim 15, wherein the antimetabolite is fluorouracil or a pharmaceutically acceptable salt thereof.

18. The pharmaceutical composition of claim 15, wherein the tubulin inhibitor is docetaxel or a pharmaceutically acceptable salt thereof.

19. The pharmaceutical composition according to claim 15, wherein the platinum agent is oxaliplatin or a pharmaceutically acceptable salt thereof.

20. The pharmaceutical composition according to claim 15, wherein the platinum agent is cisplatin or a pharmaceutically acceptable salt thereof.

21. The pharmaceutical composition of claim 15, wherein the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.

22. The pharmaceutical composition according to any one of claims 1 to 8, wherein a) an anti-EGFR antibody-drug conjugate and b) a molecularly targeted drug are used in combination, and the molecularly targeted drug is a VEGF inhibitor or an EGFR inhibitor.

23. The pharmaceutical composition of claim 22, wherein the VEGF inhibitor is bevacizumab, ramucirumab, aflibercept beta, or axitinib.

24. The pharmaceutical composition of claim 22, wherein the EGFR inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

25. The pharmaceutical composition according to any one of claims 1 to 24, wherein a) the anti-EGFR antibody-drug conjugate; and b) the immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug are contained as active ingredients in separate formulations and are administered simultaneously or asynchronously or separately.

26. A pharmaceutical composition according to any one of claims 1 to 25 for the treatment of cancer.

27. The pharmaceutical composition of claim 26, wherein the cancer is at least one selected from the group consisting of lung cancer, colorectal cancer, kidney cancer, liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, cervical cancer, head and neck cancer, bile duct cancer, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule.

28. The pharmaceutical composition of claim 27, wherein the cancer is lung cancer.

29. The pharmaceutical composition of claim 27, wherein the cancer is colon cancer.

30. The pharmaceutical composition of claim 27, wherein the cancer is breast cancer.

31. The pharmaceutical composition of claim 27, wherein the cancer is head and neck cancer.

32. The pharmaceutical composition of claim 27, wherein the cancer is renal cancer.

33. The pharmaceutical composition of claim 27, wherein the cancer is liver cancer.

34. An anti-EGFR antibody-drug conjugate for use in combination with one or more selected from an immune checkpoint inhibitor, a chemotherapeutic agent, and a molecular targeted drug for the treatment of cancer, wherein the anti-EGFR antibody-drug conjugate is represented by the following formula (I): (In the formula, m ２ represents an integer of 1 or 2, L represents a linker connecting the N297 sugar chain and D, -Lb-La-Lp-Lc-* (wherein the asterisk indicates binding to drug D, and Lb is: (In the structural formula of Lb shown above, the asterisk indicates bonding to La, and the wavy line indicates bonding to the N297 sugar chain), where La is -C(=O)-CH ２ CH ２ Lp represents -GGFG- or -GGPI-, Lc represents -NH-CH ２ -), the Ab is an anti-EGFR antibody or an antigen-binding fragment thereof comprising a heavy chain comprising a CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and a light chain comprising a CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, a CDRL2 consisting of the amino acid sequence represented by DAS, and a CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 7, wherein the N297 sugar chain is N297-(Fuc)MSG1 having the structure represented by the following formula: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of the 1-3 chain side of the branched chain of β-Man in the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1- or 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), or N297-(Fuc)SG: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), and D represents (wherein the asterisk indicates binding to L, and W represents -NH-)) 35. D. The anti-EGFR antibody-drug conjugate of claim 34, represented by the formula: (wherein the asterisk indicates binding to L).

36. The anti-EGFR antibody-drug conjugate has the following formula: (Wherein Ab, N297 sugar chain and m ２ The anti-EGFR antibody-drug conjugate of claim 34 or 35, wherein:

37. An anti-EGFR antibody-drug conjugate having the following formula: In the formula, m ２ is 2, the Ab comprises a heavy chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and a light chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2, and the N297 sugar chain is N297-(Fuc)SG having a structure represented by the following formula: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, the asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring, and n ５ The anti-EGFR antibody-drug conjugate of any one of claims 34 to 36, wherein R represents an integer of 3.

38. The anti-EGFR antibody-drug conjugate of any one of claims 34 to 37, wherein the Ab comprises a heavy chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 119 of SEQ ID NO: 1, and a light chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 107 of SEQ ID NO:

2.

39. The anti-EGFR antibody-drug conjugate of any one of claims 34 to 38, wherein the Ab comprises a heavy chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence set forth in SEQ ID NO: 1, and a light chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence set forth in SEQ ID NO:

2.

40. The anti-EGFR antibody-drug conjugate of any one of claims 34 to 39, wherein the Ab comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 1 and a light chain consisting of the amino acid sequence shown in SEQ ID NO:

2.

41. The anti-EGFR antibody-drug conjugate of any one of claims 34 to 38, wherein the Ab is comprised in an antibody or antigen-binding fragment of said antibody contained in the anti-EGFR antibody-drug conjugate of any one of claims 34 to 40, and comprises: (i) a heavy chain consisting of an amino acid sequence in which one or two amino acids are deleted from the carboxyl terminus of the amino acid sequence of the heavy chain; and (ii) a light chain.

42. The anti-EGFR antibody-drug conjugate of any one of claims 34 to 41, for use in combination with an immune checkpoint inhibitor for the treatment of cancer, wherein the immune checkpoint inhibitor is an antibody selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-TIGIT antibody, and an anti-LAG-3 antibody, an antigen-binding fragment thereof, or a compound comprising any of them.

43. The anti-EGFR antibody-drug conjugate of claim 42, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, cemiplimab, or dostarlimab.

44. The anti-EGFR antibody-drug conjugate of claim 42, wherein the anti-PD-L1 antibody is atezolizumab, duvalumab, or avelumab.

45. The anti-EGFR antibody-drug conjugate of claim 42, wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab.

46. ​​The anti-EGFR antibody-drug conjugate of claim 42, wherein the anti-TIGIT antibody is tiragolumab or vibostolimab.

47. The anti-EGFR antibody-drug conjugate of claim 42, wherein the anti-LAG-3 antibody is leratolimab.

48. The anti-EGFR antibody-drug conjugate of any one of claims 34 to 41, for use in combination with a chemotherapeutic agent for the treatment of cancer, wherein the chemotherapeutic agent is an antimetabolite, a tubulin inhibitor, an alkylating agent, a platinum compound, or a topoisomerase inhibitor.

49. The anti-EGFR antibody-drug conjugate of claim 48, wherein the antimetabolite is gemcitabine or a pharmaceutically acceptable salt thereof.

50. The anti-EGFR antibody-drug conjugate of claim 48, wherein the antimetabolite is fluorouracil or a pharmaceutically acceptable salt thereof.

51. The anti-EGFR antibody-drug conjugate of claim 48, wherein the tubulin inhibitor is docetaxel or a pharmaceutically acceptable salt thereof.

52. The anti-EGFR antibody-drug conjugate of claim 48, wherein the platinum agent is oxaliplatin or a pharmaceutically acceptable salt thereof.

53. The anti-EGFR antibody-drug conjugate of claim 48, wherein the platinum agent is cisplatin or a pharmaceutically acceptable salt thereof.

54. The anti-EGFR antibody-drug conjugate of claim 48, wherein the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.

55. An anti-EGFR antibody-drug conjugate according to any one of claims 34 to 41, for use in combination with a molecularly targeted drug for the treatment of cancer, wherein the molecularly targeted drug is a VEGF inhibitor or an EGFR inhibitor.

56. The anti-EGFR antibody-drug conjugate of claim 55, wherein the VEGF inhibitor is bevacizumab, ramucirumab, aflibercept beta, or axitinib.

57. The anti-EGFR antibody-drug conjugate of claim 55, wherein the EGFR inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

58. The anti-EGFR antibody-drug conjugate according to any one of claims 34 to 57, wherein the anti-EGFR antibody-drug conjugate; and the immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug are contained as active ingredients in separate formulations and administered simultaneously or asynchronously or separately.

59. The anti-EGFR antibody-drug conjugate of any one of claims 34 to 58, wherein the cancer is selected from the group consisting of lung cancer, colorectal cancer, kidney cancer, liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, cervical cancer, head and neck cancer, bile duct cancer, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule.

60. The anti-EGFR antibody-drug conjugate of claim 59, wherein the cancer is lung cancer.

61. The anti-EGFR antibody-drug conjugate of claim 59, wherein the cancer is colon cancer.

62. The anti-EGFR antibody-drug conjugate of claim 59, wherein the cancer is breast cancer.

63. The anti-EGFR antibody-drug conjugate of claim 59, wherein the cancer is head and neck cancer.

64. The anti-EGFR antibody-drug conjugate of claim 59, wherein the cancer is renal cancer.

65. The anti-EGFR antibody-drug conjugate of claim 59, wherein the cancer is liver cancer.

66. A method for treating a disease, comprising administering to a subject in need of disease treatment a combination of: a) an anti-EGFR antibody-drug conjugate; and b) one or more selected from an immune checkpoint inhibitor, a chemotherapeutic agent, and a molecular targeted drug, wherein the anti-EGFR antibody-drug conjugate is represented by the following formula (I): (In the formula, m ２ represents an integer of 1 or 2, L represents a linker connecting the N297 sugar chain and D, -Lb-La-Lp-Lc-* (wherein the asterisk indicates binding to drug D, and Lb is: (In the structural formula of Lb shown above, the asterisk indicates bonding to La, and the wavy line indicates bonding to the N297 sugar chain), where La is -C(=O)-CH ２ CH ２ Lp represents -GGFG- or -GGPI-, Lc represents -NH-CH ２ -), the Ab is an anti-EGFR antibody or an antigen-binding fragment thereof comprising a heavy chain comprising a CDRH1 consisting of the amino acid sequence represented by SEQ ID NO: 3, a CDRH2 consisting of the amino acid sequence represented by SEQ ID NO: 4, and a CDRH3 consisting of the amino acid sequence represented by SEQ ID NO: 5, and a light chain comprising a CDRL1 consisting of the amino acid sequence represented by SEQ ID NO: 6, a CDRL2 consisting of the amino acid sequence represented by DAS, and a CDRL3 consisting of the amino acid sequence represented by SEQ ID NO: 7, wherein the N297 sugar chain is N297-(Fuc)MSG1 having the structure represented by the following formula: (where the wavy line indicates binding to Asn297 of Ab, and L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of the 1-3 chain side of the branched chain of β-Man in the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1- or 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), or N297-(Fuc)SG: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, an asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring of Lb in the linker L, and n ５ represents an integer of 2 to 5), and D represents (wherein the asterisk indicates bonding to L, and W represents -NH-).

67. D.

67. The method of claim 66, wherein the compound is represented by the formula: (wherein the asterisk indicates that the compound is attached to L).

68. An anti-EGFR antibody-drug conjugate having the following formula: (Wherein Ab, N297 sugar chain and m ２ 68. The method of claim 66 or 67, wherein:

69. An anti-EGFR antibody-drug conjugate having the formula: In the formula, m ２ is 2, the Ab comprises a heavy chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 119th amino acid residues of SEQ ID NO: 1, and a light chain variable region consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence represented by the 1st to 107th amino acid residues of SEQ ID NO: 2, and the N297 sugar chain is N297-(Fuc)SG having a structure represented by the following formula: (wherein the wavy line indicates binding to Asn297 of Ab, L(PEG) is -(CH ２ -CH ２ -O)n ５ -CH ２ -CH ２ -NH-, the amino group at the right end of L(PEG) is amide-bonded to the carboxyl group at the 2-position of the sialic acid at the non-reducing end of both the 1-3 chain side and the 1-6 chain side of the branched chain of β-Man of the N297 sugar chain, the asterisk indicates bonding to the nitrogen atom at the 1-position or the 3-position on the 1,2,3-triazole ring, and n ５ The method according to any one of claims 66 to 68, wherein represents an integer of 3.

70. The method of any one of claims 66 to 69, wherein the Ab comprises a heavy chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 119 of SEQ ID NO: 1, and a light chain variable region consisting of the amino acid sequence represented by amino acid residues 1 to 107 of SEQ ID NO:

2.

71. The method of any one of claims 66 to 70, wherein the Ab comprises a heavy chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence set forth in SEQ ID NO:1, and a light chain consisting of an amino acid sequence that is 80% or more, preferably 90% or more, and more preferably 95% or more identical to the amino acid sequence set forth in SEQ ID NO:

2.

72. The method of any one of claims 66 to 71, wherein the Ab comprises a heavy chain consisting of the amino acid sequence set forth in SEQ ID NO:1 and a light chain consisting of the amino acid sequence set forth in SEQ ID NO:

2.

73. The method of any one of claims 66 to 70, wherein the Ab is comprised in an antibody or antigen-binding fragment of the antibody contained in the anti-EGFR antibody-drug conjugate of any one of claims 66 to 72, and comprises: (i) a heavy chain consisting of an amino acid sequence in which one or two amino acids are deleted from the carboxyl terminus of the heavy chain amino acid sequence; and (ii) a light chain.

74. A method for treating a disease, comprising administering in combination to a subject in need of disease treatment: a) an anti-EGFR antibody-drug conjugate; and b) an immune checkpoint inhibitor, wherein the immune checkpoint inhibitor is an antibody selected from the group consisting of an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, an anti-TIGIT antibody, and an anti-LAG-3 antibody, an antigen-binding fragment thereof, or a compound comprising any of them. The method of any one of claims 66 to 73.

75. The method of claim 74, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, spartalizumab, cemiplimab, or dostarlimab.

76. The method of claim 74, wherein the anti-PD-L1 antibody is atezolizumab, duvalumab, or avelumab.

77. The method of claim 74, wherein the anti-CTLA-4 antibody is ipilimumab or tremelimumab.

78. The method of claim 74, wherein the anti-TIGIT antibody is tiragolumab or vibostolimab.

79. The method of claim 74, wherein the anti-LAG-3 antibody is leratolimab.

80. A method for treating a disease, comprising administering in combination to a subject in need of disease treatment: a) an anti-EGFR antibody-drug conjugate; and b) a chemotherapeutic agent, wherein the chemotherapeutic agent is an antimetabolite, a tubulin inhibitor, an alkylating agent, a platinum compound, or a topoisomerase inhibitor, according to any one of claims 66 to 73.

81. The method of claim 80, wherein the antimetabolite is gemcitabine or a pharmaceutically acceptable salt thereof.

82. The method of claim 80, wherein the antimetabolite is fluorouracil or a pharmaceutically acceptable salt thereof.

83. The method of claim 80, wherein the tubulin inhibitor is docetaxel or a pharmaceutically acceptable salt thereof.

84. The method of claim 80, wherein the platinum agent is oxaliplatin or a pharmaceutically acceptable salt thereof.

85. The method of claim 80, wherein the platinum agent is cisplatin or a pharmaceutically acceptable salt thereof.

86. The method of claim 80, wherein the topoisomerase inhibitor is irinotecan or a pharmaceutically acceptable salt thereof.

87. A method for treating a disease, comprising administering a) an anti-EGFR antibody-drug conjugate; and b) a molecular targeted drug in combination to a subject in need of disease treatment, wherein the molecular targeted drug is a VEGF inhibitor or an EGFR inhibitor, according to any one of claims 66 to 73.

88. The method of claim 87, wherein the VEGF inhibitor is bevacizumab, ramucirumab, aflibercept beta, or axitinib.

89. The method of claim 87, wherein the EGFR inhibitor is osimertinib or a pharmaceutically acceptable salt thereof.

90. The method according to any one of claims 66 to 89, wherein the anti-EGFR antibody-drug conjugate; and the immune checkpoint inhibitor, chemotherapeutic agent, or molecular targeted drug are contained as active ingredients in separate formulations and administered simultaneously or asynchronously or separately.

91. The method of any one of claims 66 to 90, wherein the disease is cancer.

92. The method of claim 91, wherein the cancer is selected from the group consisting of lung cancer, colorectal cancer, kidney cancer, liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, breast cancer, bladder cancer, gastric cancer, esophageal cancer, uterine cancer, cervical cancer, head and neck cancer, bile duct cancer, myeloma, sarcoma, EGFR mutation-positive cancer, and cancer with a mutation in an EGFR downstream signaling molecule.

93. The method of claim 92, wherein the cancer is lung cancer.

94. The method of claim 92, wherein the cancer is colon cancer.

95. The method of claim 92, wherein the cancer is breast cancer.

96. The method of claim 92, wherein the cancer is head and neck cancer.

97. The method of claim 92, wherein the cancer is renal cancer.

98. The method of claim 92, wherein the cancer is liver cancer.

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