Antibody drug complexes comprising toll-like receptor 7 / 8 dual agonist compounds

Abstract

The present invention addresses the problem of developing an antibody-drug complex containing a TLR7 / 8 dual agonist compound or a salt thereof, and providing a pharmaceutical composition for use in the prevention or treatment of various cancers. The present inventors have discovered that a compound having a TLR7 / 8 dual agonist activity or a salt thereof, studies an antibody drug complex that can be an active ingredient of a pharmaceutical composition for preventing or treating various cancers, and discovers that the antibody drug complex or a salt thereof according to the present invention has a TNF-[alpha] and INF-[gamma]-producing effect, an in vivo anti-tumor effect, and an in vivo anti-tumor effect. It has been clear that it can be used in the prevention or treatment of cancer. In addition, an anti-CLDN6 antibody for use in the antibody-drug complex of the present invention or a salt thereof has been found. An antibody drug complex comprising the TLR7 / 8 dual agonist compound of the present invention or a salt thereof, and the compound of the present invention or a salt thereof in formula (II) have an effect of producing TNF-alpha and INF-gamma and an in vivo antitumor effect, and are expected to be used as a prophylactic or therapeutic agent for cancer.

Description

Technical Field

[0001] The present invention relates to an antibody-drug conjugate or a salt thereof containing a compound (hereinafter, the compound is also referred to as a drug) having a Toll-like receptor 7 and 8 dual agonist (hereinafter, also referred to as a TLR7 / 8 dual agonist) action, and the TLR7 / 8 dual agonist compound or a salt thereof. In addition, the present invention relates to an anti-CLDN6 antibody for use in the antibody-drug conjugate or a salt thereof. Background Art

[0002] TLR binds to pathogen-associated molecular patterns of bacteria, fungi, protozoa, and viruses, and functions as the first line of defense against invading pathogens. TLR7 and 8, which are included in the TLR family, are known to be pattern recognition receptors that are mainly expressed in dendritic cells, macrophages, monocytes, B cells, etc. Activation of TLR7 and 8 leads to antigen uptake and maturation of dendritic cells, and an increase in T cell stimulation ability.

[0003] Patent document 1 describes TM-Ln-AM (wherein TM is a targeting moiety, L is a linker, n is an integer selectable from 0 and 1, and AM is an activating moiety capable of activating dendritic cells, NK cells, tumor cells, or a combination thereof) as a compound capable of activating TLR7 and / or TLR8. For example, a compound comprising trastuzumab and resiquimod, which are known to bind to human epidermal growth factor receptor 2 (hereinafter also referred to as Her2), is disclosed.

[0004] In addition, the structural formula of resiquimod is shown below for reference.

[0005] [Chemical formula 1]

[0006]

[0007] In addition, Patent Documents 2 and 3 report Her2 antibody-drug complexes containing a TLR7 agonist.

[0008]

[0009] (For the symbols in the formula, refer to Patent Documents 2 and 3.)

[0010] In addition, Patent Documents 4 and 5 report on FOLR1 antibody-drug complexes containing a TLR7 agonist.

[0011] [Chemical formula 3]

[0012]

[0013] (For the symbols in the formula, refer to Patent Documents 4 and 5.)

[0014] In addition, Patent Document 6 reports a TLR7 agonist compound.

[0015] [Chemical formula 4]

[0016]

[0017] (Refer to Patent Document 6 for the symbols in the formula. 3 H, -L2C(=O)OR 7 、-C(=O)OL6R 12 、-C(=O)OL2R 12 、-L2C(=O)OL2R 12 、-L4C(=O)OL5OH、-L4R 12 , (omitted), L 4 -(CH2) m -,(omitted)R 12 a)-N(R 11 ) 2, b) unsubstituted 5-6 membered heterocycloalkyl having 1-2 heteroatoms independently selected from N and O, c) 5-6 membered heterocycloalkyl substituted with =O and having 1-2 heteroatoms independently selected from N and O, d) substituted with C1-C3 alkyl or -C(=O)OR 7 substituted 5-6 membered heterocycloalkyl having 1-2 heteroatoms independently selected from N and O, or e) unsubstituted phenyl, m is independently selected from 1, 2, 3 and 4.

[0018] In addition, Patent Document 4 also reports a TLR7 agonist compound.

[0019] [Chemical formula 5]

[0020]

[0021] (Refer to Patent Document 4 for the symbols in the formula. 3 Each is independently -N(R 3a )2.-OR 3b ,-C(R 3c )2NH2、C 1-6 Alkyl, heterocycloalkyl, heteroaryl, partially saturated heteroaryl, or two R 3 is bonded to the same carbon atom and becomes one with the bonded carbon atom to form a spiroheterocycloalkyl group, wherein R 3 The heterocycloalkyl, spiroheterocycloalkyl, heteroaryl, and spiroheterocycloalkyl groups contain 1, 2, 3, or 4 heteroatoms selected from N, S, and O, and may be replaced by 1 to 2 C1-3 Alkyl substitution.)

[0022] Claudin 6 (hereinafter referred to as CLDN6) is a member of the Claudin family of proteins. It is a 4-transmembrane-type membrane protein consisting of 220 amino acid residues. It is an oncofetal gene expressed in embryoid bodies associated with the fate of mouse and human stem cells and epithelial cells. The expression level of CLDN6 in normal tissues is very low. On the other hand, it is overexpressed in tumors including childhood brain tumors, germ cell tumors, and cancers such as gastric adenocarcinoma and ovarian cancer, suggesting that it can be used as a target for cancer treatment (International Journal of Molecular Science 2021, vol. 22, p. 13416).

[0023] As anti-CLDN6 antibodies, GT512muMAB 64A (Patent Document 7) and AE3-20 (Patent Document 8) have been reported. As cancer therapeutic drugs, there have been no reports of antibody-drug complexes containing TLR7 and / or TLR8 agonist compounds targeting CLDN6.

[0024] Prior art literature

[0025] Patent Literature

[0026] Patent Document 1: International Publication No. 2014 / 012479

[0027] Patent Document 2: International Publication No. 2017 / 072662

[0028] Patent Document 3: International Publication No. 2018 / 198091

[0029] Patent Document 4: International Publication No. 2020 / 252043

[0030] Patent Document 5: International Publication No. 2020 / 252015

[0031] Patent Document 6: International Publication No. 2015 / 168279

[0032] Patent Document 7: International Publication No. 2011 / 057788

[0033] Patent Document 8: International Publication No. 2009 / 087978 Summary of the invention

[0034] Problems to be solved by the invention

[0035] The present invention aims to create a compound or a salt thereof having TLR7 / 8 dual agonist activity, develop an antibody-drug conjugate containing the TLR7 / 8 dual agonist compound, and provide a pharmaceutical composition for the prevention or treatment of various cancers such as ovarian cancer, testicular cancer, cervical cancer, and lung cancer. In addition, an anti-CLDN6 antibody for use in an antibody-drug conjugate or a salt thereof is provided.

[0036] Means for solving problems

[0037] The present inventors conducted in-depth research on the antibody-drug complex of formula (I), and found that the drug of formula (II) of the present invention has TLR7 / 8 dual agonist effect, the antibody used in the antibody-drug complex of formula (I) shows binding activity to the target (human CLDN6), and the antibody-drug complex of formula (I) induces the production of TNF-α and INF-γ, and further found an in vivo anti-tumor effect, thereby completing the present invention.

[0038] That is, the present invention relates to a pharmaceutical composition comprising the following [1] to [3] and an excipient.

[0039] [1] An antibody-drug conjugate represented by formula (I) or a salt thereof:

[0040] [Chemical formula 6]

[0041]

[0042] (Where,

[0043] Ab is an antibody or its antigen-binding fragment,

[0044] X is S or NH,

[0045] R 1 is a lower alkyl group or -CH2-isoxazolediyl-CH3,

[0046] Here, in R 1 In the case of -CH2-isoxazolediyl-CH3, X is S,

[0047] R 2 is halogen or H,

[0048] R 3 is CH3 or H,

[0049] R 4 is a group represented by formula (a), formula (b) or formula (c),

[0050] [Chemical formula 7]

[0051]

[0052] Ring A is a 4- to 6-membered cyclic amine consisting of multiple Cs and one N.

[0053] Ring B is a 4- to 6-membered cyclic amine composed of multiple Cs, 1 N and 1 G,

[0054] G is N or CH,

[0055] R A is -CH2O-, -C(CH3)2O-, -O- or -CH2NH-,

[0056] Here, in R A When it is -O- or -CH2NH-, X is S; R 2 is halogen; or R 3 For CH3,

[0057] R B is a halogenated alkyl, CH2OH, C(CH3)2OH, OH, CH2NH2 or H,

[0058] Here, in R B When OH, CH2NH2 or H is present, G is CH; X is S; R 2 is halogen; or R 3 For CH3,

[0059] R C C 3-6 Cycloalkyl, or -CH2-C 3-6 Cycloalkyl,

[0060] L is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, -(CH2CH2O) m -C(=O)-lower alkylene, or -C(=O)-cyclohexanediyl-lower alkylene-,

[0061] m is an integer from 1 to 10,

[0062] n is 1 to 16).

[0063] [2] A compound of formula (II) or a salt thereof:

[0064] [Chemical formula 8]

[0065]

[0066] (Where,

[0067] X is S or NH,

[0068] R 1is a lower alkyl group or -CH2-isoxazolediyl-CH3,

[0069] Here, in R 1 In the case of -CH2-isoxazolediyl-CH3, X is S,

[0070] R 2 is halogen or H,

[0071] R 3 is CH3 or H,

[0072] R 40 is a group represented by formula (a1), formula (b1) or formula (c1),

[0073] [Chemical formula 9]

[0074]

[0075] Ring A is a 4- to 6-membered cyclic amine composed of multiple C atoms and one N atom.

[0076] Ring B is a 4- to 6-membered cyclic amine composed of multiple Cs, 1 N and 1 G,

[0077] G is N or CH,

[0078] R A is -CH2O-, -C(CH3)2O-, -O-, or -CH2NH-,

[0079] Here, in R A When it is -O- or -CH2NH-, X is S; R 2 is halogen; or R 3 For CH3,

[0080] R B is a haloalkyl, CH2OH, C(CH3)2OH, OH, CH2NH2, or H,

[0081] Here, in R B When OH, CH2NH2 or H is present, G is CH; X is S; R 2 is halogen; or R 3 For CH3,

[0082] R C C 3-6 Cycloalkyl or -CH2-C 3-6 Cycloalkyl,

[0083] M is H, or a group represented by formula (d):

[0084] [Chemical formula 10]

[0085]

[0086] L D -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, -(CH2CH2O) m -C(=O)-lower alkylene-, or -C(=O)-cyclohexanediyl-lower alkylene-,

[0087] m is an integer from 1 to 10).

[0088] [3] An anti-CLDN6 antibody or an antigen-binding fragment thereof, which is an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 2, and a light chain variable region consisting of amino acids 1 to 108 of SEQ ID NO: 4; or an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 6, and a light chain variable region consisting of amino acids 1 to 108 of SEQ ID NO: 8.

[0089] In addition, the present invention is not limited to the above-mentioned embodiments [1] to [3], and also includes embodiments in which the contents described in the detailed invention of the specification are appropriately combined. As a certain embodiment, the present invention may also be a compound of formula (II) described in [2] above or a salt thereof wherein R 40 A compound or a salt thereof in which M of a group represented by formula (a1), formula (b1) or formula (c1) is H. In addition, as a certain embodiment, the present invention may also be a compound of formula (II) described in [2] above or a salt thereof in which R 40 An antibody-drug conjugate or a salt thereof in which an antibody is bound to the position M of the group represented by formula (a1), formula (b1), or formula (c1) via a linker. It should be noted that the binding group in the linker that binds to the antibody is not limited to the maleimide group included in the group represented by formula (d) described in [2] above, and various binding groups described below can be appropriately used. In addition, the linker part is not limited to the L included in the group represented by formula (d) described in [2] above. D , various non-cleaving linkers and various cleaving linkers described later can be used appropriately.

[0090] In addition, the present invention relates to an antibody-drug conjugate of formula (I) or a salt thereof; and a pharmaceutical composition comprising an antibody-drug conjugate of formula (I) or a salt thereof and one or more pharmaceutically acceptable excipients, in particular a pharmaceutical composition for the prevention and / or treatment of cancer. The pharmaceutical composition comprises a cancer prevention and / or treatment agent containing an antibody-drug conjugate of formula (I) or a salt thereof.

[0091] The present invention relates to the use of an antibody-drug conjugate of formula (I) or a salt thereof for producing a pharmaceutical composition for preventing and / or treating cancer, the use of an antibody-drug conjugate of formula (I) or a salt thereof for preventing and / or treating cancer, an antibody-drug conjugate of formula (I) or a salt thereof for use in preventing and / or treating cancer, and a method for preventing and / or treating cancer comprising administering an effective amount of an antibody-drug conjugate of formula (I) or a salt thereof to a subject.

[0092] In addition, the present invention relates to a compound of formula (II) or a salt thereof; and a pharmaceutical composition comprising a compound of formula (II) or a salt thereof and one or more pharmaceutically acceptable excipients, in particular a pharmaceutical composition for the prevention and / or treatment of cancer. The pharmaceutical composition comprises a cancer prevention and / or treatment agent containing a compound of formula (II) or a salt thereof.

[0093] The present invention relates to the use of a compound of formula (II) or a salt thereof for producing a pharmaceutical composition for preventing and / or treating cancer, the use of a compound of formula (II) or a salt thereof for preventing and / or treating cancer, a compound of formula (II) or a salt thereof for use in preventing and / or treating cancer, and a method for preventing and / or treating cancer comprising administering an effective amount of a compound of formula (II) or a salt thereof to a subject.

[0094] In addition, the present invention relates to a pharmaceutical composition comprising an antibody-drug conjugate of a compound of formula (II) or a salt thereof or a salt thereof and one or more pharmaceutically acceptable excipients, in particular a pharmaceutical composition for the prevention and / or treatment of cancer. The pharmaceutical composition comprises a cancer prevention and / or treatment agent, which contains an antibody-drug conjugate of a compound of formula (II) or a salt thereof or a salt thereof.

[0095] The present invention relates to the use of an antibody-drug conjugate or a salt thereof containing a compound of formula (II) or a salt thereof for producing a pharmaceutical composition for the prevention and / or treatment of cancer, the use of an antibody-drug conjugate or a salt thereof for use in the prevention and / or treatment of cancer, an antibody-drug conjugate or a salt thereof for use in the prevention and / or treatment of cancer, and a method for the prevention and / or treatment of cancer comprising administering an effective amount of an antibody-drug conjugate or a salt thereof containing a compound of formula (II) or a salt thereof to a subject.

[0096] It should be noted that the "subject" refers to a human or other animal that needs the prevention or treatment, and in a certain aspect, refers to a human that needs the prevention or treatment.

[0097] In the present invention, the type of cancer is not particularly limited, and may be any of solid cancers and blood cancers. Examples include various diffuse peritoneal cancers, gastric cancer, lung cancer, acute lymphoblastic leukemia, acute myeloid leukemia, Hodgkin lymphoma, Blood cancers such as non-Hodgkin lymphoma, B-cell lymphoma, multiple myeloma, and T-cell lymphoma; solid cancers such as myelodysplastic syndrome, adenocarcinoma, squamous cell carcinoma, adenosquamous cell carcinoma, undifferentiated carcinoma, large cell carcinoma, non-small cell lung cancer, small cell lung cancer, mesothelioma, skin cancer, cutaneous T-cell lymphoma, breast cancer, prostate cancer, bladder cancer, vaginal cancer, neck cancer, head and neck cancer, uterine cancer, cervical cancer, liver cancer, gallbladder cancer, bile duct cancer, kidney cancer, pancreatic cancer, colon cancer, large intestine cancer, rectal cancer, small intestine cancer, stomach cancer, esophageal cancer, testicular cancer, ovarian cancer, and brain tumors; and solid cancers such as chondrosarcoma, Ewing's sarcoma, and brain tumors, except for cancers of bone tissue, cartilage tissue, adipose tissue, muscle tissue, vascular tissue, and hematopoietic tissue. Sarcoma), malignant hemangioendothelioma, malignant Schwannoma, osteosarcoma, soft tissue sarcoma and other sarcomas; glioblastoma, glioblastoma multiforme, hepatoblastoma, medulloblastoma, nephroblastoma, neuroblastoma, pancreatic blastoma, pleuropulmonary blastoma or omental blastoma and other blastomas, etc. In one embodiment, ovarian cancer, testicular cancer, cervical cancer or lung cancer can be listed. In one embodiment, ovarian cancer expressing CLDN6, testicular cancer expressing CLDN6, cervical cancer expressing CLDN6, or lung cancer expressing CLDN6 can be listed.

[0098] This specification includes the disclosure of Japanese Patent Application No. 2022-174538 which is the basis of priority of this application.

[0099] Effects of the Invention

[0100] The compound of formula (II) or its salt has TLR7 / 8 dual agonist effect. The compound of formula (II) or its salt, or the antibody-drug complex of formula (I) or its salt containing the TLR7 / 8 dual agonist compound has the effect of producing TNF-α and INF-γ, binds to tumor-associated antigens expressed on the tumor surface to kill cancer cells, and also has an in vivo anti-tumor effect, and can therefore be used as a preventive and / or therapeutic agent for cancer.

[0101] In addition, the anti-CLDN6 antibody of the present invention has binding activity to human CLDN6 and can be used as an antibody-drug conjugate or a salt thereof for use as a preventive and / or therapeutic agent for cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0102] [ Figure 1 ] Figure 1 The results of evaluating the binding activity of antibodies Ab0, Ab1, and Ab2 to the gastric cancer cell line NUGC-3 expressing human CLDN6 by flow cytometry are shown. The vertical axis represents the geometric mean fluorescence intensity, and the horizontal axis represents the antibody concentration (ng / mL).

[0103] [ Figure 2 ] Figure 2 The results of measuring the cytokine production levels of each of the antibody-drug complexes ExL1A1 and ExL1A2 are shown. The vertical axis represents the production level of TNF-α or INF-γ cytokines (pg / mL), and the horizontal axis represents the antibody concentration (nM).

[0104] [ Figure 3 ] Figure 3 The results of in vivo activity evaluation of antibody-drug complexes ExL1A1, ExL5A1, and ExL7A1 in xenograft models are shown. The vertical axis represents tumor volume (mm 3 ), the horizontal axis represents the number of days (days) after the first administration of the antibody. The significant probability P value was obtained by comparing the tumor volume of the PBS-administered group and the tumor volume of the ExL1A1, ExL5A1, and ExL7A1-administered groups 14 days later using an unpaired t-test (*: P < 0.05, **: P < 0.01).

[0105] [ Figure 4 ] Figure 4 The results of measuring the cytokine production of the antibody-drug complex ExL1A1 when the medium was added or when the TLR8 inhibitor CU-CPT9a and the TLR7 / 8 inhibitor Enpatorn were added are shown. The vertical axis represents the production of INF-γ cytokines (pg / mL) and the horizontal axis represents the antibody concentration (nM). DETAILED DESCRIPTION

[0106] Hereinafter, the present invention will be described in detail.

[0107] In this specification, unless otherwise specified, the following terms have the following meanings. The following definitions are intended to clarify the defined terms and are not intended to be limiting. When a term used herein is not specifically defined, the term is used with the meaning generally accepted by those skilled in the art. When other chemical formulas in this specification also use a symbol in a chemical formula, the same symbol represents the same meaning unless otherwise specified.

[0108] "Lower alkyl" refers to a linear or branched chain having 1 to 6 carbon atoms (hereinafter referred to as C 1-6 ) alkyl, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc. As a certain embodiment, the lower alkyl is C 1-6 Alkyl, as a certain embodiment, lower alkyl is C 1-5 In one embodiment, the alkyl group is a methyl group, and in another embodiment, the alkyl group is a n-pentyl group.

[0109] "Lower alkylene" refers to a straight or branched chain C 1-6 The lower alkylene group is C 1-4 In one embodiment, the alkylene group is an ethylene group (-CH2H2-).

[0110] "Halogen" refers to F, Cl, Br or I.

[0111] "Haloalkyl" is a lower alkyl group substituted with one or more halogens. In one embodiment, the haloalkyl group is a C 1-6 Alkyl. In one embodiment, the haloalkyl group is a lower alkyl group substituted with 1 to 5 halogens, in one embodiment, the haloalkyl group is a lower alkyl group substituted with 1 to 5 F groups, in one embodiment, the haloalkyl group is a methyl group substituted with 1 to 3 F groups, and in one embodiment, the haloalkyl group is a monofluoromethyl group.

[0112] “C 3-6 "Cycloalkyl" is C 3-6 For example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. As a certain embodiment, C 3-6 Cycloalkyl is C 3-4 Cycloalkyl, as a certain form, C 3-6 Cycloalkyl is cyclobutyl. In one embodiment, C 3-6 Cycloalkyl is cyclopropyl.

[0113] "Cyclic amine" is a non-aromatic cyclic amine having at least one nitrogen atom (N) and other elements constituting the ring being carbon atoms (C). Examples include aziridine, azetidine, pyrrolidine, piperidine, piperazine, azepan, diazepan, and the like. If it is a divalent group, it is aziridinediyl, azetidinediyl, pyrrolidinyl, piperidinediyl, piperazinediyl, azepandiyl, diazepandiyl, and the like. If it is a monovalent group, it is aziridine, azetidine, pyrrolidinyl, piperidinyl, piperazinyl, azepandiyl, diazepandiyl, and the like. In a certain embodiment, it is a 4-6 membered cyclic amine composed of multiple C and 1 N. In a certain embodiment, it is a 4-6 membered cyclic amine composed of multiple C, 1 N, and 1 G, and G is N or CH. In one embodiment, it is a 5-membered cyclic amine composed of 4 Cs and 1 N. In one embodiment, it is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, and G is N or CH. In one embodiment, it is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, and G is N. In one embodiment, it is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, and G is CH.

[0114] "4- to 6-membered cyclic amine" refers to a cyclic amine with a 4- to 6-membered ring. Examples include azetidine, pyrrolidine, piperidine, and piperazine. If it is a divalent group, it is azetidinediyl, pyrrolidinediyl, piperidinediyl, and piperazinediyl. If it is a monovalent group, it is azetidineyl, pyrrolidinyl, piperidinyl, and piperazineyl. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine, in one embodiment, the 4- to 6-membered cyclic amine is a 5-membered cyclic amine, in one embodiment, the 4- to 6-membered cyclic amine is a piperidinediyl or piperazinediyl, in one embodiment, the 4- to 6-membered cyclic amine is a pyrrolidinediyl or piperazinediyl, in one embodiment, the 4- to 6-membered cyclic amine is a piperidinediyl, in one embodiment, the 4- to 6-membered cyclic amine is a pyrrolidinediyl, and in one embodiment, the 4- to 6-membered cyclic amine is a piperazinediyl. In one embodiment, the 4- to 6-membered cyclic amine is a 4- to 6-membered cyclic amine composed of a plurality of carbon atoms and a nitrogen atom. In one embodiment, the 4- to 6-membered cyclic amine is a 4- to 6-membered cyclic amine composed of a plurality of carbon atoms, a nitrogen atom, and a G, wherein G is CH or N. In one embodiment, the 4- to 6-membered cyclic amine is a 5-membered cyclic amine composed of 4 C and 1 N. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine composed of 5 C and 1 N. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine composed of 4 C, 1 N, and 1 G, wherein G is CH or N. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine composed of 4 C, 1 N, and 1 G, wherein G is N. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine composed of 4 C, 1 N, and 1 G, wherein G is CH. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine composed of 4 Cs, 1 N, and 1 G, wherein G is CH or N. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs, and 1 G, wherein G is CH or N. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs, and 1 G, wherein G is N. In one embodiment, the 4- to 6-membered cyclic amine is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs, and 1 G, wherein G is CH.

[0115] "Antibody drug conjugate" refers to a complex formed by binding an antibody and at least one drug via a linker. The drug can be delivered to the cell or tissue targeted by the antibody. As a certain embodiment, the antibody drug conjugate is a complex formed by binding an antibody and a drug via a non-cutting linker. "Antibody drug conjugate containing a compound" in this specification refers to an antibody drug conjugate formed by binding an antibody to a compound via a linker. As a certain embodiment, it is an antibody drug conjugate with a linker, and as a certain embodiment, it is an antibody drug conjugate without a linker.

[0116] "Linker" is a chemical group that binds an antibody to a drug and can form at least two covalent bonds. There are non-cleavable linkers and cleavable linkers. As linkers used in antibody drug conjugates, for example, alkylene, polyethylene glycol (PEG), or peptides can be listed. In order to bind or combine linkers to each other, amide groups, ether groups, carbamate groups, etc. can be used. As binding groups that bind to antibodies in linkers, for example, maleimide groups, pyridyl disulfide groups, isocyanate groups, etc. can be listed.

[0117] "Non-cleavable linker" refers to a linker that is not degraded even by acidic conditions in lysosomes or specific proteases in cells. For example, -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, -(CH2CH2O) m -C(=O)lower alkylene-, or -C(=O)-cyclohexanediyl-CH2-, "m" is an integer from 1 to 10, and combinations of these embodiments that do not cause contradictions. In one embodiment, the non-cleaving linker is -CH2CH2-, -C(=O)CH2CH2-, -C(=O)NHCH2CH2-, -C(=O)CH2CH2-(OCH2CH2) m -NH-C(=O)CH2CH2-、-(CH2CH2O) m -C(=O)CH2CH2-, or -C(=O)-cyclohexanediyl-CH2-. In one embodiment, the non-cleaving linker is -CH2CH2-, -C(=O)CH2CH2-, or -C(=O)NHCH2CH2-. In one embodiment, the non-cleaving linker is -C(=O)CH2CH2-, or -C(=O)NHCH2CH2-. In one embodiment, the non-cleaving linker is -C(=O)CH2CH2-. In one embodiment, when the non-cleaving linker is -C(=O)NHCH2CH2-, the oxygen atom (O) of the compound or its salt is bonded to the -C(=O) of the linker. In one embodiment, "m" is an integer of 1 to 8, or an integer of 4 to 8, and in one embodiment, "m" is 4 or 8.

[0118] "Cutable linker" refers to a linker that can be selectively cut by acidic conditions in lysosomes, endogenous enzymes such as specific protease nucleases and peptidases in cells. As such specific examples, for example, hydrazone bonds, SS bonds, peptide bonds, ester bonds, phosphate bonds, phosphodiester bonds, ester bonds of one or both, carbamate bonds, etc. can be listed. For example, Val-Cit (valine-citrulline), Val-Ala, Ala-Ala, Ala-Ala-Ala, Gly-Gly-Phe-Gly, Val-Cit-PAPC (valine-citrulline-p-benzyl alcohol), Val-Ala-PABC, or, and combinations of these methods that do not produce contradictions (Bioconjugate Chemistry 2002, vol.13, p.855-869. Protein & Cell, 2018, vol.9, p.33-46).

[0119] "Drug-linker complex" is a compound or salt thereof in which a drug is covalently bonded to a linker. As a certain embodiment, it is a compound or salt thereof in which a compound of the present invention is covalently bonded to a linker. The drug-linker complex can be combined with an antibody or an antibody-binding fragment thereof to produce an antibody-drug complex. Here, the linker of the drug-linker complex is covalently bonded to the antibody. Here, in the antibody-drug complex, at least one drug-linker complex is combined with an antibody or an antibody-binding fragment thereof. As a certain embodiment, at least one drug (anticancer agent or marker, etc.)-linker complex can be combined with the antibody portion of the antibody-drug complex. As anticancer agents, monomethyl auristatin E (MMAE) or monomethyl auristatin F (MMAF) as microtubule polymerization inhibitors, mitotic inhibitors such as maytansine (DM1) and the like can be listed. As markers, radioactive isotopes (RI-labeled antibodies) and the like can be listed.

[0120] "TLR7 / 8 dual agonist" refers to a TLR7 agonist and a TLR8 agonist. In one embodiment, the EC of TLR7 in the reporter assay is 50 The value is stronger than 150nM (i.e., EC 50 value is less than 150 nM), and the EC 50 The value is stronger than 15 μM (i.e., EC 50 A compound or a salt thereof having an agonist effect having an EC value of less than 15 μM. In one embodiment, the EC value of the compound or salt thereof for TLR7 in a reporter assay is 50 Values ​​greater than 1 μM (i.e., EC 50 value is less than 1 μM), and for TLR8 EC 50A compound or a salt thereof having an agonist effect with an EC50 value stronger than 15 μM (ie, an EC50 value less than 15 μM). A "TLR7 / 8 dual agonist compound" refers to a compound or a salt thereof that is a TLR7 / 8 dual agonist.

[0121] "TLR7 / 8 dual inhibitor" refers to a TLR7 inhibitor and a TLR8 inhibitor. For example, Enpatoran, etc.

[0122] The antibody portion of an "antibody-drug conjugate" is an antibody or antigen-binding fragment that binds to a specific antigen.

[0123] There are five types of antibodies: IgG, IgM, IgA, IgD, and IgE. The basic structure of antibody molecules is common to all types, consisting of a heavy chain with a molecular weight of 50,000 to 70,000 and a light chain with a molecular weight of 20,000 to 30,000. The heavy chain is composed of a polypeptide chain that usually contains about 440 amino acids, and each type has a characteristic structure, corresponding to IgG, IgM, IgA, IgD, and IgE, and is called Igγ, Igμ, Igα, Igδ, and Igε. In addition, there are IgG1, IgG2, IgG3, and IgG4 subtypes in IgG, and the corresponding heavy chains are called Igγ1, Igγ2, Igγ3, and Igγ4. The light chain is composed of a polypeptide chain that usually contains about 220 amino acids, and two types are known, λ type and κ type, called Igλ and Igκ, respectively. The basic structure of an antibody molecule consists of two identical heavy chains and two identical light chains bound together by disulfide bonds (SS bonds) and non-covalent bonds, with a molecular weight of 150,000 to 190,000. Two light chains can be paired with any heavy chain. Each antibody molecule is always composed of two identical light chains and two identical heavy chains.

[0124] In antibodies, there are 4 intrachain SS bonds in the heavy chain (5 in Igμ and Igε) and 2 in the light chain. Every 100 to 110 amino acid residues form a ring. The three-dimensional structure is similar between each ring and is called a structural unit or domain. The domain located at the N-terminus of both the heavy and light chains has an invariant amino acid sequence even if it is an antibody of the same type (subtype) produced by the same animal. It is called a variable region, and each domain is called a heavy chain variable region (VH) and a light chain variable region (VL). The amino acid sequence closer to the C-terminus than the variable region is roughly constant in each type or subtype and is called a constant region (each domain is represented as CH1, CH2, CH3 or CL).

[0125] In antibodies, the antigenic determinant is composed of VH and VL, and the specificity of binding depends on the amino acid sequence of this site. On the other hand, biological activities such as binding to complement and various Fc receptor expressing cells reflect the structural differences of the constant regions of various types of Ig. It is known that the variability of the light chain and heavy chain variable regions is basically limited to 3 small hypervariable regions present in any chain, and these regions are called complementary determining regions (CDR; CDR1, CDR2, CDR3 from the N-terminal side). The rest of the variable region is called the framework region (FR), which is relatively constant.

[0126] "Antigen-binding fragment" is a fragment that contains VH and VL of an antibody and has binding activity to an antigen. For example, as such representative antigen-binding fragments, single-chain variable region fragments (scFv), Fab, Fab', and F(ab')2 can be cited. scFv is a monovalent antibody fragment consisting of VH and VL connected by a peptide linker. Fab is a monovalent antibody fragment consisting of a heavy chain fragment containing a light chain and a part of VH, a CH1 domain, and a hinge region. Fab' is a monovalent antibody fragment consisting of a heavy chain fragment containing a light chain and a part of VH, a CH1 domain, and a hinge region, and contains a cysteine ​​residue constituting an SS bond between heavy chains in the hinge region. The F(ab')2 fragment is a bivalent antibody fragment formed by two Fab' fragments bound by an SS bond between heavy chains in the hinge region. As a certain embodiment, the antigen-binding fragment used in the present invention is scFv, Fab, Fab', or F(ab')2.

[0127] "Post-translational modification" refers to chemical or biological modification after mRNA translation when the antibody is expressed in a cell. Chemical modifications include the combination of a chemical part with an amino acid backbone, a chemical modification of an N-bond or O-bond hydrocarbon chain, and the like. Biological modifications include: a modified body obtained by adding a methionine residue to the N-terminus by using a prokaryotic host cell to express a substance that undergoes post-translational modification (e.g., addition of a sugar chain to an N-bond or O-bond, processing of the N-terminus or C-terminus, deamidation, isomerization of aspartic acid, oxidation of methionine), and the like. It is known in the art that such post-translational modification does not affect the activity of the antibody (Anal Biochem., 2006, Vol. 348, p. 24-39).

[0128] "Anti-TAA antibody" refers to an antibody that binds to a tumor-associated antigen (TAA) expressed on the surface of tumor cells. Anti-TAA antibodies are antibodies or antigen-binding fragments thereof that can label tumor cells. Examples of anti-TAA antibodies include anti-A33 antibodies, anti-B7-H3 antibodies, anti-CanAg antibodies, anti-CD20 antibodies, anti-CD22 antibodies, anti-CD30 antibodies, anti-CD33 antibodies, anti-CD56 antibodies, anti-CD70 antibodies, anti-CEA bodies, anti-CLDN6 antibodies, anti-EphA2 antibodies, anti-G250 antibodies, anti-MUC1 antibodies, anti-GPNMB antibodies, anti-Integrin antibodies, PSMA antibodies, anti-Tenascin-C antibodies, anti-TROP-2 antibodies, anti-TSPAN8 antibodies, etc., but are not limited thereto.

[0129] "Anti-CLDN6 antibody" refers to an antibody or an antigen-binding fragment thereof that binds to CLDN6. In one embodiment, the anti-CLDN6 antibody is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the following group consisting of (Ab-A) to (Ab-D).

[0130] (Ab-A) an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acids 31 to 35 of SEQ ID NO: 2, a CDR2 consisting of an amino acid sequence of amino acids 50 to 65 of SEQ ID NO: 2, and a CDR3 consisting of an amino acid sequence of amino acids 95 to 102 of SEQ ID NO: 2, and the light chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acids 24 to 34 of SEQ ID NO: 4, a CDR2 consisting of an amino acid sequence of amino acids 50 to 56 of SEQ ID NO: 4, and a CDR3 consisting of an amino acid sequence of amino acids 89 to 97 of SEQ ID NO: 4;

[0131] (Ab-B) an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acids 31 to 35 of SEQ ID NO: 6, a CDR2 consisting of an amino acid sequence of amino acids 50 to 65 of SEQ ID NO: 6, and a CDR3 consisting of an amino acid sequence of amino acids 95 to 102 of SEQ ID NO: 6, and the light chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acids 24 to 34 of SEQ ID NO: 8, a CDR2 consisting of an amino acid sequence of amino acids 50 to 56 of SEQ ID NO: 8, and a CDR3 consisting of an amino acid sequence of amino acids 89 to 97 of SEQ ID NO: 8;

[0132] (Ab-C) an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of the amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 2 and a light chain variable region consisting of the amino acid sequence of amino acids 1 to 108 of SEQ ID NO: 4; and

[0133] (Ab-D) An anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of the amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 6 and a light chain variable region consisting of the amino acid sequence of amino acids 1 to 108 of SEQ ID NO: 8.

[0134] As a certain embodiment, the anti-CLDN6 antibody is an antibody that can further comprise a heavy chain constant region and a light chain constant region. As the constant region, any subtype of constant region can be selected (for example, Igγ1, Igγ2, Igγ3 or Igγ4 as a heavy chain, and a constant region of Igλ or Igκ as a light chain), the heavy chain constant region can be a human Igγ1 constant region, and the light chain constant region can be a human Igκ constant region.

[0135] In one embodiment, the anti-CLDN6 antibody is Ab1 or Ab2.

[0136] “Ab1” is an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 and a light chain consisting of the amino acid sequence of SEQ ID NO: 4,

[0137] “Ab2” is an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO:6 and a light chain consisting of the amino acid sequence of SEQ ID NO:8.

[0138] In one embodiment, the anti-CLDN6 antibody is a post-translationally modified anti-CLDN6 antibody or an antigen-binding fragment thereof. In one embodiment, it is an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 and a light chain consisting of the amino acid sequence of SEQ ID NO: 4, or an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 6 and a light chain consisting of the amino acid sequence of SEQ ID NO: 8. In one embodiment, the anti-CLDN6 antibody is a post-translationally modified Ab1 or Ab2 antibody.

[0139] The binding activity of the antibody to tumor cells can be confirmed using a known binding activity assay method. As a method for measuring binding activity, for example, known methods such as enzyme-linked immunosorbent assay (ELISA) can be cited. In the case of using ELISA, for example, CLDN6 is immobilized on an ELISA plate, and after adding the test antibody to it to react, a secondary antibody such as an anti-IgG antibody labeled with an enzyme such as horseradish peroxidase (HRP) is reacted. After the reaction and washing, the binding of the secondary antibody is identified by an activity assay using a reagent that detects its activity (for example, in the case of HRP labeling, TMB Microwell Peroxidase Substrate (Kirkegaard & Perry Laboratories, 50-76-03)), so that it can be confirmed whether the test antibody binds to CLDN6.

[0140] (Aspects of the present invention [1])

[0141] One embodiment of the antibody-drug conjugate of formula (I) of the present invention or a salt thereof is shown below. It should be noted that all embodiments can be freely combined. Even if the combination is not specifically described, one or more embodiments can be combined into a certain embodiment.

[0142] (1) In one embodiment, the formula (I) is an antibody-drug conjugate of the formula (IA) or a salt thereof:

[0143] [Chemical formula 11]

[0144]

[0145] In one embodiment, formula (I) is an antibody-drug conjugate or a salt thereof of formula (IA) bound to SH of antibody Ab.

[0146] (2) In a certain embodiment, Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-tumor-associated antigen (TAA) antibody or an antigen-binding fragment thereof. In a certain embodiment, Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-TAA antibody or a salt thereof selected from the group consisting of anti-A33 antibody, anti-B7-H3 antibody, anti-CanAg antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CEA body, anti-CLDN6 antibody, anti-EphA2 antibody, anti-G250 antibody, anti-MUC1 antibody, anti-GPNMB antibody, anti-Integrin antibody, antibody PSMA antibody, anti-Tenascin-C antibody, anti-TROP-2 antibody, and anti-TSPAN8 antibody. In a certain embodiment, Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-CLDN6 antibody or an antigen-binding fragment thereof.In one embodiment, Ab of the antibody-drug conjugate or a salt thereof of formula (I) is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the following group consisting of (Ab-A) to (Ab-D): (Ab-A) an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acid numbers 31 to 35 of SEQ ID NO: 2, a CDR2 consisting of an amino acid sequence of amino acid numbers 50 to 65 of SEQ ID NO: 2, and a CDR3 consisting of an amino acid sequence of amino acid numbers 50 to 65 of SEQ ID NO: 2. The light chain variable region comprises: a CDR1 composed of an amino acid sequence of amino acid numbers 24 to 34 of SEQ ID NO: 4, a CDR2 composed of an amino acid sequence of amino acid numbers 50 to 56 of SEQ ID NO: 4, and a CDR3 composed of an amino acid sequence of amino acid numbers 89 to 97 of SEQ ID NO: 4; (Ab-B) an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: an amino acid sequence of amino acid numbers 6 The light chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acid numbers 24 to 34 of sequence number 8, a CDR2 consisting of an amino acid sequence of amino acid numbers 50 to 56 of sequence number 8, and a CDR3 consisting of an amino acid sequence of amino acid numbers 89 to 97 of sequence number 8. ; (Ab-C) an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acid numbers 1 to 117 of SEQ ID NO: 2, and a light chain variable region consisting of an amino acid sequence of amino acid numbers 1 to 108 of SEQ ID NO: 4; and (Ab-D) an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acid numbers 1 to 117 of SEQ ID NO: 6, and a light chain variable region consisting of an amino acid sequence of amino acid numbers 1 to 108 of SEQ ID NO: 8. In one embodiment, Ab of the antibody-drug conjugate or a salt thereof of formula (I) is an anti-CLDN6 antibody or an antigen-binding fragment thereof of (Ab-A). In one embodiment, Ab of the antibody-drug conjugate or a salt thereof of formula (I) is an anti-CLDN6 antibody or an antigen-binding fragment thereof of (Ab-B). In one embodiment, Ab of the antibody-drug conjugate or a salt thereof of formula (I) is an anti-CLDN6 antibody or an antigen-binding fragment thereof of (Ab-C). In one embodiment, Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-CLDN6 antibody or an antigen-binding fragment thereof of (Ab-D). In one embodiment, Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of Ab-A and Ab-B.As a certain embodiment, Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of Ab-C and Ab-D. As a certain embodiment, Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-CLDN6 antibody selected from the following Ab1 and Ab2: (Ab1) an anti-CLDN6 antibody comprising a heavy chain consisting of an amino acid sequence of sequence number 2 and a light chain consisting of an amino acid sequence of sequence number 4; and (Ab2) an anti-CLDN6 antibody comprising a heavy chain consisting of an amino acid sequence of sequence number 6 and a light chain consisting of an amino acid sequence of sequence number 8. Ab is an antibody-drug conjugate of formula (I) or a salt thereof in which Ab is an anti-CLDN6 antibody Ab1. Ab is an antibody-drug conjugate of formula (I) or a salt thereof in which Ab is an anti-CLDN6 antibody Ab2. As a certain embodiment, Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-CLDN6 antibody of Ab1 or Ab2 that has been post-translationally modified. In one embodiment, the Ab of the antibody-drug conjugate of formula (I) or a salt thereof is an anti-CLDN6 antibody as follows: an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 which is post-translationally modified and a light chain consisting of the amino acid sequence of SEQ ID NO: 4; or an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 6 which is post-translationally modified and a light chain consisting of the amino acid sequence of SEQ ID NO: 8.

[0147] (3) In one embodiment, X in the antibody-drug conjugate of formula (I) or a salt thereof is S. In one embodiment, X in the antibody-drug conjugate of formula (I) or a salt thereof is NH.

[0148] (4) In one embodiment, R of the antibody-drug conjugate of formula (I) or a salt thereof is 1 In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 1 In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 1 is -CH2-isoxazolediyl-CH3, and X is S. In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 1 is -CH2-(5-methylisoxazol-3-yl), and X is S.

[0149] (5) In one embodiment, R of the antibody-drug conjugate of formula (I) or a salt thereof is 2 In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 2 is Br, Cl or H. In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 2 is Cl or H. In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 2 For H.

[0150] (6) In one embodiment, R of the antibody-drug conjugate of formula (I) or a salt thereof is 3 In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 3 For H.

[0151] (7) In one embodiment, R of the antibody-drug conjugate of formula (I) or a salt thereof is 4 It is a group represented by formula (a), formula (b) or formula (c).

[0152] [Chemical formula 12]

[0153]

[0154] In one embodiment, R of the antibody-drug conjugate of formula (I) or a salt thereof is 4 is a group represented by formula (a) or formula (b). In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 4 is a group represented by formula (b) or formula (c). In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 4 is a group represented by formula (c) or formula (a). In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 4 is a group represented by formula (a). In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 4 is a group represented by formula (b). In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 4 It is a group represented by formula (c).

[0155] (8) In one embodiment, Ring A of the antibody-drug conjugate of formula (I) or a salt thereof is a 5-membered cyclic amine consisting of 4 Cs and 1 N. In one embodiment, Ring A of the antibody-drug conjugate of formula (I) or a salt thereof is a 6-membered cyclic amine consisting of 5 Cs and 1 N. In one embodiment, Ring A of the antibody-drug conjugate of formula (I) or a salt thereof is pyrrolidine. In one embodiment, Ring A of the antibody-drug conjugate of formula (I) or a salt thereof is piperazine.

[0156] (9) In one embodiment, Ring B of the antibody-drug conjugate of formula (I) or a salt thereof is a 6-membered cyclic amine consisting of 4 Cs, 1 N, and 1 G. In one embodiment, Ring B of the antibody-drug conjugate of formula (I) or a salt thereof is a 6-membered cyclic amine consisting of 2 Cs, 1 N, 2 Cs, and 1 G. In one embodiment, Ring B of the antibody-drug conjugate of formula (I) or a salt thereof is piperazine.

[0157] (10) In one embodiment, G of the antibody-drug conjugate of formula (I) or a salt thereof is N. In one embodiment, G of the antibody-drug conjugate of formula (I) or a salt thereof is CH.

[0158] (11) In one embodiment, R of the antibody-drug conjugate of formula (I) or a salt thereof is A is -CH2O- or -C(CH3)2O-. In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is A It is -CH2O-.

[0159] (12) In one embodiment, R of the antibody-drug conjugate of formula (I) or a salt thereof is B is a halogenated alkyl group, CH2OH or C(CH3)2OH. In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is B is a halogenated alkyl group or H, where R B When it is H, G is N. In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is B In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is B In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is B It is monofluoromethyl.

[0160] (13) In one embodiment, R of the antibody-drug conjugate of formula (I) or a salt thereof is C C 3-6 In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is C -CH2-C 3-6 In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is C In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is C It is -CH2-cyclobutyl.

[0161] (14) In one embodiment, L of the antibody-drug conjugate of formula (I) or a salt thereof is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m-NH-C(=O)lower alkylene-. In one embodiment, L of the antibody-drug conjugate of formula (I) or a salt thereof is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-. In one embodiment, L of the antibody-drug conjugate of formula (I) or a salt thereof is -C(=O)lower alkylene-, or -C(=O)NHlower alkylene-. In one embodiment, L of the antibody-drug conjugate of formula (I) or a salt thereof is -C(=O)lower alkylene-. In one embodiment, L of the antibody-drug conjugate of formula (I) or a salt thereof is a non-cleaving linker. In one embodiment, L of the antibody-drug conjugate of formula (I) or a salt thereof is -C(=O)CH2CH2-, or -C(=O)NHCH2CH2-. In one embodiment, L of the antibody-drug conjugate of formula (I) or a salt thereof is -C(=O)CH2CH2-. In one embodiment, L of the antibody-drug conjugate of formula (I) or a salt thereof is -C(=O)NHCH2CH2-. In the embodiment described in (14), when L of the antibody-drug conjugate of formula (I) or a salt thereof comprises -C(=O)NH lower alkylene- (here, as a subordinate concept thereof, for example, -C(=O)NHCH2CH2- is also included), -C(=O)NH lower alkylene- and R 4 R of formula (a) A Combined, R A In the embodiment described in (14), when L of the antibody drug conjugate of formula (I) or a salt thereof comprises -C(=O)NH lower alkylene- (here, as a subordinate concept, for example, -C(=O)NHCH2CH2- is also included), -C(=O)NH lower alkylene- and R 4 R of formula (a) A Combined, RA is -CH2O-.

[0162] (15) In one embodiment, m of the antibody-drug conjugate of formula (I) or a salt thereof is an integer of 4 to 8. In one embodiment, m of the antibody-drug conjugate of formula (I) or a salt thereof is an integer of 4 or 8.

[0163] (16) In one embodiment, n of the antibody-drug conjugate of formula (I) or a salt thereof is 1 to 16. In one embodiment, n of the antibody-drug conjugate of formula (I) or a salt thereof is 2 to 5. In one embodiment, n of the antibody-drug conjugate of formula (I) or a salt thereof is 1 to 8.

[0164] (17) An antibody-drug conjugate or a salt thereof of formula (I) in which two or more of the embodiments described in (1) to (16) above are combined without causing any inconsistency. An antibody-drug conjugate or a salt thereof of formula (I) in which two or more of the embodiments described in (1) to (16) are combined without causing any inconsistency, an antibody embodiment of (2) in which a certain embodiment of the antibody selected as an antibody of an antibody-drug conjugate or a salt thereof is selected, a drug embodiment of (3) to (13) in which a certain embodiment of the drug selected as an antibody-drug conjugate or a salt thereof is selected, and a linker embodiment of (14) and (15) in which a certain embodiment of the linker selected as an antibody-drug conjugate or a salt thereof is selected.

[0165] As examples of the combination described in (17) above, the following examples can be cited.

[0166] [1] The antibody-drug conjugate of formula (I) or a salt thereof, wherein Ab is an anti-TAA antibody or an antigen-binding fragment thereof selected from the group consisting of anti-A33 antibody, anti-B7-H3 antibody, anti-CanAg antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CEA antibody, anti-CLDN6 antibody, anti-EphA2 antibody, anti-G250 antibody, anti-MUC1 antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-TROP-2 antibody, and anti-TSPAN8 antibody, and X is S or NH, and R 1 is a lower alkyl group or -CH2-isoxazolediyl-CH3 (here, R 1 In the case of -CH2-isoxazolediyl-CH3, X is S), R 2 is halogen or H, R 3 CH3 or H, R 4 is a group represented by formula (a), formula (b) or formula (c),

[0167] [Chemical formula 13]

[0168]

[0169] Ring A is a 4- to 6-membered cyclic amine composed of multiple Cs and 1 N, and Ring B is a 4- to 6-membered cyclic amine composed of multiple Cs, 1 N, and 1 G, where G is N or CH, and R A is -CH2O-, -C(CH3)2O-, -O- or -CH2NH-, where R A When it is -O- or -CH2NH-, X is S; R 2 is halogen; or R 3 For CH3, R Bis a halogenated alkyl, CH2OH, C(CH3)2OH, OH, CH2NH2 or H, wherein R B When OH, CH2NH2 or H is present, G is CH; X is S; R 2 is halogen; or R 3 For CH3, R C C 3-6 Cycloalkyl, or -CH2-C 3-6 Cycloalkyl, L is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, -(CH2CH2O) m -C(=O)-lower alkylene- or -C(=O)-cyclohexanediyl-lower alkylene-, wherein m is an integer of 1 to 10 and n is 1 to 16.

[0170] [1a] An antibody-drug conjugate of formula (I) or a salt thereof, wherein Ab is an anti-CLDN6 antibody selected from the following Ab1 and Ab2: (Ab1) an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 and a light chain consisting of the amino acid sequence of SEQ ID NO: 4, and (Ab2) an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 6 and a light chain consisting of the amino acid sequence of SEQ ID NO: 8, X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 The formula is (a), (b) or (c), wherein ring A is a 5-membered cyclic amine composed of 4 Cs and 1 N, and ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, wherein G is N, and R A -CH2-O-, R B is a halogenated alkyl group, R C C 3-6 Cycloalkyl, L is -C(=O)CH2CH2-, or -C(=O)NHCH2CH2-.

[0171] [2] The antibody-drug conjugate or a salt thereof of formula (I), wherein ring A is a 5-membered cyclic amine consisting of 4 Cs and 1 N, and ring B is a 6-membered cyclic amine consisting of 2 Cs, 1 N, 2 Cs and 1 G, and G is N.

[0172] [3] The antibody-drug conjugate of formula (I) or a salt thereof according to [2], wherein R A is -CH2O- or -C(CH3)2O-, R B It is halogenated alkyl, CH2OH or C(CH3)2OH.

[0173] [4] The antibody-drug conjugate of formula (I) or a salt thereof according to [3], wherein X is NH, R 1 is a lower alkyl group, R 3 For H.

[0174] [5] The antibody-drug conjugate of formula (I) or a salt thereof according to [4], wherein L is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, or -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, m is an integer of 4-8.

[0175] [6] The antibody-drug conjugate or a salt thereof of formula (I) according to [5], wherein L is -lower alkylene-, -C(=O)lower alkylene-, or -C(=O)NHlower alkylene-.

[0176] [7] the antibody-drug conjugate of formula (I) or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 is halogen, R 3 For H, R 4 Formula (a), Ring A is a 5-membered cyclic amine consisting of 4 Cs and 1 N, R A is -CH2O-, and L is -C(=O)NHCH2CH2-.

[0177] [8] the antibody-drug conjugate of formula (I) or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 It is a 6-membered cyclic amine of formula (b) consisting of 2 Cs, 1 N, 2 Cs and 1 G, wherein G is N, R B is a halogenated alkyl group, and L is -C(=O)CH2CH2-.

[0178] [9] the antibody-drug conjugate of formula (I) or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 Formula (c), R C C 3-6 Cycloalkyl, L is -C(=O)CH2CH2-.

[0179]

[10] The antibody-drug conjugate of formula (I) or a salt thereof according to [1], wherein the antibody-drug conjugate is selected from the group consisting of the following antibody-drug conjugates:

[0180] Antibody-drug complex formed by binding Ab to 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazine-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione,

[0181] An antibody-drug complex formed by binding Ab to [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamic acid {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl ester,

[0182] An antibody-drug complex in which Ab is conjugated with N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamide, and

[0183] The antibody-drug complex is formed by binding Ab to 1-{3-[(3R)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione.

[0184]

[11] The antibody-drug conjugate of formula (I) or a salt thereof according to [2] to

[10] , wherein Ab is an anti-TAA antibody or an antigen-binding fragment thereof of any of the following: anti-A33 antibody, anti-B7-H3 antibody, anti-CanAg antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CEA antibody, anti-CLDN6 antibody, anti-EphA2 antibody, anti-G250 antibody, anti-MUC1 antibody, anti-GPNMB antibody, anti-Integrin antibody, PSMA antibody, anti-Tenascin-C antibody, anti-TROP-2 antibody, anti-TSPAN8 antibody.

[0185]

[12] The antibody-drug conjugate or a salt thereof according to

[11] , wherein Ab is an anti-CLDN6 antibody.

[0186]

[13] The antibody-drug conjugate or a salt thereof according to

[12] , wherein Ab is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of Ab-A and Ab-B; Ab-A is an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acids 31 to 35 of SEQ ID NO: 2, a CDR2 consisting of an amino acid sequence of amino acids 50 to 65 of SEQ ID NO: 2, and a CDR3 consisting of an amino acid sequence of amino acids 95 to 102 of SEQ ID NO: 2, and the light chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acids 24 to 34 of SEQ ID NO: 4, a CDR2 consisting of an amino acid sequence of amino acids 50 to 56 of SEQ ID NO: 4, and a CDR4 consisting of an amino acid sequence of amino acids 95 to 102 of SEQ ID NO: 2. and a CDR3 consisting of an amino acid sequence of amino acids 89 to 97 of sequence number 4; Ab-B is an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acids 31 to 35 of sequence number 6, a CDR2 consisting of an amino acid sequence of amino acids 50 to 65 of sequence number 6, and a CDR3 consisting of an amino acid sequence of amino acids 95 to 102 of sequence number 6, and the light chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acids 24 to 34 of sequence number 8, a CDR2 consisting of an amino acid sequence of amino acids 50 to 56 of sequence number 8, and a CDR3 consisting of an amino acid sequence of amino acids 89 to 97 of sequence number 8.

[0187]

[14] The antibody-drug conjugate or a salt thereof according to

[12] , wherein Ab is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of Ab-C and Ab-D; Ab-C is an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 2, and a light chain variable region consisting of an amino acid sequence of amino acids 1 to 108 of SEQ ID NO: 4; and Ab-D is an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 6, and a light chain variable region consisting of an amino acid sequence of amino acids 1 to 108 of SEQ ID NO: 8.

[0188]

[15] The antibody-drug conjugate or a salt thereof according to

[12] , wherein Ab is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of Ab1 and Ab2; Ab1 is an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 and a light chain consisting of the amino acid sequence of SEQ ID NO: 4; and Ab2 is an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 6 and a light chain consisting of the amino acid sequence of SEQ ID NO: 8.

[0189]

[16] The antibody-drug conjugate or a salt thereof of formula (I), wherein Ab is the anti-CLDN6 antibody Ab1, X is NH, R 1 is a lower alkyl group, R 2 is halogen, R 3 For H, R 4 Formula (a), Ring A is a 5-membered cyclic amine consisting of 4 Cs and 1 N, R A is -CH2-O-, and L is -C(=O)NHCH2CH2-.

[0190]

[17] The antibody-drug conjugate of formula (I) or a salt thereof, wherein Ab is the anti-CLDN6 antibody Ab1, X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 Formula (b), Ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, G is N, R B is a halogenated alkyl group, and L is -C(=O)CH2CH2-.

[0191]

[18] The antibody-drug conjugate or a salt thereof of formula (I), wherein Ab is the anti-CLDN6 antibody Ab1, X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 Formula (c), R C C 3-6 Cycloalkyl, L is -C(=O)CH2CH2- or -C(=O)NHCH2CH2-.

[0192] Specific examples of the antibody-drug conjugate or a salt thereof included in the present invention include the following.

[0193] An antibody-drug complex or a salt thereof in which Ab1 is bonded to 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione.

[0194] An antibody-drug complex or a salt thereof formed by binding Ab2 to 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione.

[0195] An antibody-drug complex or a salt thereof in which Ab1 is bound to [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamic acid {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl ester.

[0196] An antibody-drug complex or a salt thereof formed by binding Ab1 to N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamide.

[0197] An antibody-drug complex or a salt thereof in which Ab1 is bonded to 1-{3-[(3R)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione.

[0198] Specific examples of the antibody-drug conjugate of formula (I) or a salt thereof included in the present invention include the following.

[0199] [Table 1]

[0200]

[0201] Ab1 is an antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 and a light chain consisting of the amino acid sequence of SEQ ID NO: 4,

[0202] Ab2 is an antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 6 and a light chain consisting of the amino acid sequence of SEQ ID NO: 8.

[0203] n is 1 to 16. In one embodiment, n is 2 to 5.

[0204] (Aspects of the present invention [2])

[0205] One embodiment of the compound of formula (II) or a salt thereof of the present invention is shown below.

[0206] (C1) In one embodiment, X in the compound of formula (II) or a salt thereof is S. In one embodiment, X in the compound of formula (II) or a salt thereof is NH.

[0207] (C2) In one embodiment, R of the compound of formula (II) or a salt thereof is 1is a lower alkyl or -CH2-isoxazolediyl-CH3, where R 1 When it is -CH2-isoxazolediyl-CH3, X is S. In one embodiment, R of the compound of formula (II) or a salt thereof 1 As one embodiment, R of the compound of formula (II) or a salt thereof is 1 As one embodiment, R of the compound of formula (II) or a salt thereof is 1 is -CH2-isoxazolediyl-CH3, and X is S. In one embodiment, R of the antibody-drug conjugate or a salt thereof of formula (I) is 1 is -CH2-(5-methylisoxazol-3-yl), and X is S.

[0208] (C3) In one embodiment, R of the compound of formula (II) or a salt thereof is 2 The compound of formula (II) or its salt R 2 is Br or H. In one embodiment, R of the compound of formula (II) or a salt thereof is 2 is Cl or H. In one embodiment, R of the compound of formula (II) or a salt thereof 2 For H.

[0209] (C4) In one embodiment, R of the compound of formula (II) or a salt thereof is 3 As one embodiment, R of the compound of formula (II) or a salt thereof is 3 For H.

[0210] (C5) In one embodiment, the compound of formula (II) or a salt thereof has R 4 is a group represented by formula (a1), formula (b1) or formula (c1):

[0211] [Chemical formula 14]

[0212]

[0213] In one embodiment, R of the compound of formula (II) or a salt thereof is 40 is a group represented by formula (a1) or formula (b1). In one embodiment, R of the compound of formula (II) or a salt thereof is 40 is a group represented by formula (a1). In one embodiment, R of the compound of formula (II) or a salt thereof is 40 is a group represented by formula (b1). In one embodiment, R of the compound of formula (II) or a salt thereof is 40 It is a group represented by formula (c1).

[0214] (C6) In one embodiment, the ring A of the compound of formula (II) or a salt thereof is a 5-membered cyclic amine consisting of 4 Cs and 1 N. In one embodiment, the ring A of the compound of formula (II) or a salt thereof is pyrrolidine. In one embodiment, the ring A of the compound of formula (II) or a salt thereof is a 6-membered cyclic amine consisting of 5 Cs and 1 N.

[0215] (C7) In one embodiment, the ring B of the compound of formula (II) or a salt thereof is a 6-membered cyclic amine consisting of 4 Cs, 1 N, and 1 G, and G is N. In one embodiment, the ring B of the compound of formula (II) or a salt thereof is a 6-membered cyclic amine consisting of 2 Cs, 1 N, 2 Cs, and 1 G. In one embodiment, the ring B of the compound of formula (II) or a salt thereof is a 6-membered cyclic amine consisting of 2 Cs, 1 N, 2 Cs, and 1 G, and G is N or CH. In one embodiment, the ring B of the compound of formula (II) or a salt thereof is a 6-membered cyclic amine consisting of 2 Cs, 1 N, 2 Cs, and 1 G, and G is N. In one embodiment, the ring B of the compound of formula (II) or a salt thereof is piperazine.

[0216] (C8) In one embodiment, G of the compound of formula (II) or a salt thereof is N. In one embodiment, G of the compound of formula (II) or a salt thereof is CH.

[0217] (C9) In one embodiment, R of the compound of formula (II) or a salt thereof is A is -CH2O- or -C(CH3)2O-. In one embodiment, R of the compound of formula (II) or a salt thereof is A It is -CH2O-.

[0218] (C10) In one embodiment, the compound of formula (II) or a salt thereof has R B is a halogenated alkyl group, CH2OH or C(CH3)2OH. In one embodiment, R of the compound of formula (II) or a salt thereof is B is a halogenated alkyl group or H, where R B When it is H, G is CH. In one embodiment, R of the compound of formula (II) or a salt thereof B In one embodiment, R of the compound of formula (II) or a salt thereof is B It is monofluoromethyl.

[0219] (C11) In one embodiment, R of the compound of formula (II) or a salt thereof is C C 3-6 In one embodiment, R of the compound of formula (II) or a salt thereof is C -CH2-C 3-6 In one embodiment, R of the compound of formula (II) or a salt thereof is CIn one embodiment, R of the compound of formula (II) or a salt thereof is C It is -CH2-cyclobutyl.

[0220] (C12) In one embodiment, M of the compound of formula (II) or a salt thereof is H. In one embodiment, M of the compound of formula (II) or a salt thereof is a group represented by formula (d):

[0221] [Chemical formula 15]

[0222]

[0223] (C13) In one embodiment, L of the compound of formula (II) or a salt thereof is D -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-. In one embodiment, L of the compound of formula (II) or a salt thereof is D is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-. In one embodiment, L of the compound of formula (II) or a salt thereof is D is -C(=O)lower alkylene- or -C(=O)NHlower alkylene-. In one embodiment, L of the compound of formula (II) or a salt thereof is D is -C(=O)lower alkylene-. In one embodiment, L of the compound of formula (II) or a salt thereof is D As a certain embodiment, L of the compound of formula (II) or a salt thereof is D is -C(=O)NH lower alkylene-. In one embodiment, L of the compound of formula (II) or a salt thereof is D In one embodiment, in the embodiment described in (C13), L of the compound of formula (II) or a salt thereof is D In the case of -C(=O)NH lower alkylene- (here, as a subordinate concept, for example, -C(=O)NHCH2CH2-, etc. are also included), -C(=O)NH lower alkylene- and R 40 R in formula (a1) A Combined, R A is -CH2O- or -C(CH3)2O-. In the embodiment described in (C13), L of the compound of formula (II) or its salt D In the case of -C(=O)NH lower alkylene- (here, as a subordinate concept, for example, -C(=O)NHCH2CH2-, etc. are also included), -C(=O)NH lower alkylene- and R 40R in formula (a1) A Combined, R A It is -CH2O-.

[0224] (C14) In one embodiment, m of the compound of formula (II) or a salt thereof is an integer of 4 to 8. In one embodiment, m of the compound of formula (II) or a salt thereof is an integer of 4 or 8.

[0225] (C15) A compound of formula (II) or a salt thereof for use in producing an antibody-drug conjugate of formula (I) or a salt thereof.

[0226] (C16) A compound of formula (II) or a salt thereof in which two or more of the groups described in (C1) to (C15) above are combined without causing any inconsistency.

[0227] Examples of the combination described in (C16) include the following.

[0228] [C1] A compound of formula (II) or a salt thereof, wherein X is S or NH, R 1 is a lower alkyl or -CH2-isoxazolediyl-CH3, where R 1 In the case of -CH2-isoxazolediyl-CH3, X is S, R 2 is halogen or H, R 3 CH3 or H, R 40 is a group represented by formula (a1), formula (b1) or formula (c1), ring A is a 5-membered cyclic amine composed of 4 Cs and 1 N, ring B is a 6-membered cyclic amine composed of 4 Cs, 1 N and 1 G, G is N or CH, R A is -CH2O-, -C(CH3)2O-, -O- or -CH2NH-, where R A When it is -O- or -CH2NH-, X is S; R 2 is halogen; or R 3 For CH3, R B is a halogenated alkyl, CH2OH, C(CH3)2OH, OH, CH2NH2 or H, wherein R B When OH, CH2NH2 or H is present, G is CH; X is S; R 2 is halogen; or R 3 For CH3, R C C 3-6 Cycloalkyl or -CH2-C 3-6 Cycloalkyl, M is H or a group represented by formula (d), L D -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m-NH-C(=O)lower alkylene-, -(CH2CH2O) m -C(=O)-lower alkylene- or -C(=O)-cyclohexanediyl-lower alkylene-, wherein m is an integer of 1 to 10.

[0229] [C1a] A compound of formula (II) or a salt thereof, wherein ring A is a 5-membered cyclic amine consisting of 4 Cs and 1 N, and ring B is a 6-membered cyclic amine consisting of 2 Cs, 1 N, 2 Cs and 1 G, and G is N.

[0230] [C2] A compound of formula (II) according to [C1] or [C1a] or a salt thereof, wherein R A is -CH2O- or -C(CH3)2O-, R B It is halogenated alkyl, CH2OH, or C(CH3)2OH.

[0231] [C3] A compound of formula (II) according to [C2] or a salt thereof, L D is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, or -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, m is an integer of 4-8.

[0232] [C4] A compound of formula (II) according to [C2] or a salt thereof, L D It is -C(=O)lower alkylene- or -C(=O)NHlower alkylene-.

[0233] [C5] A compound of formula (II) according to [C3] or [C4] or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 is halogen or H, R 3 For H.

[0234] [C6] A compound of formula (II) or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 is Cl or H, R 3 For H, R 40 is the formula (a1), the formula (b1) or the formula (c1), wherein ring A is a 5-membered cyclic amine composed of 4 Cs and 1 N, and ring B is a 6-membered cyclic amine composed of 4 Cs, 1 N and 1 G, G is N, and R A is -CH2O-, R B is a halogenated alkyl group, R C C 3-6 cycloalkyl, M is a group represented by formula (d), L D It is -C(=O)CH2CH2-.

[0235] [C6a] A compound of formula (II) according to [C5] or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 is Cl or H, R 3 For H, R 40 is the formula (a1), the formula (b1) or the formula (c1), wherein Ring A is a 5-membered cyclic amine composed of 4 Cs and 1 N, or a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, wherein G is N, and R A is -CH2O-, R B is a halogenated alkyl group, R C C 3-6 cycloalkyl, M is a group represented by formula (d), L D It is -C(=O)CH2CH2-.

[0236] [C7] A compound of formula (II) according to [C6a] or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 40 Formula (b1), Ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, G is N, R B is a halogenated alkyl group, L D It is -C(=O)CH2CH2-.

[0237] [C8] A compound of formula (II) or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 is halogen, R 3 For H, R 40 Formula (a1), Ring A is a 5-membered cyclic amine consisting of 4 Cs and 1 N, R A -CH2O-, L D It is -C(=O)NHCH2CH2-.

[0238] [C9] A compound of formula (II) or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 40 Formula (c), R C C 3-6 Cycloalkyl, L D It is -C(=O)CH2CH2-.

[0239] [C10] A compound of formula (II) or a salt thereof, wherein X is NH, R 1 is a lower alkyl group, R 2 is Cl or H, R3 For H, R 40 is the formula (a1), the formula (b1) or the formula (c1), wherein ring A is a 5-membered cyclic amine composed of 4 Cs and 1 N, and ring B is a 6-membered cyclic amine composed of 4 Cs, 1 N and 1 G, G is N, and R A is -CH2O-, R B is a halogenated alkyl group, R C C 3-6 cycloalkyl, M is a group represented by formula (d), L D is -C(=O)CH2CH2- or -C(=O)NHCH2CH2-, where L D In the case of -C(=O)NHCH2CH2-, -C(=O)NHCH2CH2- and R 40 R in formula (a1) A Combined, R A It is -CH2O-.

[0240] Specific examples of the compounds or salts thereof included in the present invention include the following.

[0241] 5-[(4-{[(2S)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof,

[0242] 5-[(4-{[(3R)-3-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof,

[0243] 5-({4-[(cyclopropylamino)methyl]-2-methoxyphenyl}methyl)-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof,

[0244] {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methanol or a salt thereof,

[0245] 5-({2-methoxy-4-[(piperidin-4-yl)methyl]phenyl}methyl)-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof,

[0246] 5-[(4-{[(2R)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof,

[0247] 5-[(4-{[(cyclobutylmethyl)amino]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof,

[0248] 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione or a salt thereof,

[0249] {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamate or a salt thereof,

[0250] N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propaneamide or a salt thereof,

[0251] 1-{3-[(3R)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione or a salt thereof.

[0252] Specific examples of the compounds or salts thereof included in the present invention include the following.

[0253] 5-[(4-{[(2S)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0254] 5-[(4-{[(3R)-3-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4-pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (trifluoroacetic acid) salt,

[0255] 5-({4-[(cyclopropylamino)methyl]-2-methoxyphenyl}methyl)-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0256] {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methanol,

[0257] 5-({2-methoxy-4-[(piperidin-4-yl)methyl]phenyl}methyl)-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0258] 5-[(4-{[(2R)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0259] 5-[(4-{[(cyclobutylmethyl)amino]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0260] 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione (trifluoroacetic acid) salt,

[0261] {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamate,

[0262] N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propaneamide (trifluoroacetic acid) salt,

[0263] 1-{3-[(3R)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione (trifluoroacetic acid) salt.

[0264] Examples of the compound represented by the formula (II) or a salt thereof represented by (C16) wherein M is H and included in the present invention include the following.

[0265] 5-[(4-{[(2S)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0266] 5-[(4-{[(3R)-3-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (trifluoroacetic acid) salt,

[0267] 5-({4-[(cyclopropylamino)methyl]-2-methoxyphenyl}methyl)-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0268] {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methanol,

[0269] 5-({2-methoxy-4-[(piperidin-4-yl)methyl]phenyl}methyl)-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0270] 5-[(4-{[(2R)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine,

[0271] 5-[(4-{[(cyclobutylmethyl)amino]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine.

[0272] In addition, one embodiment of the compound of formula (II) or a salt thereof of the present invention is shown below.

[0273] (C17) In the compound of formula (II) or a salt thereof, R 40 A compound or a salt thereof, wherein M of the group represented by formula (a1), formula (b1) or formula (c1) is H.

[0274] (C18) In the compound of formula (II) or a salt thereof, R 40 An antibody-drug conjugate or a salt thereof in which an antibody is bound to the position M of the group represented by formula (a1), formula (b1) or formula (c1) via a linker.

[0275] (C19) The antibody-drug conjugate or a salt thereof according to (C18), wherein the antibody is an anti-TAA antibody or an antigen-binding fragment thereof.

[0276] (C20) The antibody-drug conjugate or a salt thereof according to (C18), wherein the antibody is an anti-CLDN6 antibody or an antigen-binding fragment thereof.

[0277] The antibody-drug conjugate of formula (I) or a salt thereof can be provided by combining a compound of formula (II) or a salt thereof, any anti-TAA antibody, and any non-cleaving linker.

[0278] The antibody-drug conjugate of formula (I) or its salt, and the compound of formula (II) or its salt may exist as tautomers and geometric isomers depending on the type of substituents. In the present specification, the antibody-drug conjugate of formula (I) or its salt is sometimes described in the form of only one isomer, but the present invention also includes other isomers, and also includes substances obtained by isomer separation, or mixtures thereof.

[0279] In addition, the antibody drug conjugate of formula (I) or its salt, and the compound of formula (II) or its salt sometimes have asymmetric centers or axial asymmetry, and enantiomers (optical isomers) based on this may exist. The antibody drug conjugate of formula (I) or its salt also includes any of the enantiomers such as the (R) body and the (S) body after separation, and their mixtures (including racemic mixtures or non-racemic mixtures). In a certain embodiment, the enantiomer is "stereochemically pure". "Stereochemically pure" means that a person skilled in the art can recognize that it is substantially stereochemically pure. As a certain embodiment, the enantiomer is a compound having a stereochemical purity of, for example, 90% ee (enantiomeric excess, enantiomeric excess) or more, 95% ee or more, 98% ee or more, or 99% ee or more.

[0280] In addition, the salt of the antibody-drug conjugate of formula (I) and the salt of the compound of formula (II) refer to pharmaceutically acceptable salts of the antibody-drug conjugate of formula (I), and may form acid addition salts depending on the type of substituents. Specifically, examples include: acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, and phosphoric acid; and with organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, and glutamic acid.

[0281] In addition, the present invention also includes the antibody-drug conjugate of formula (I) and its salt or its salt, and various hydrates, solvates and polymorphs of the compound of formula (II) or its salt.

[0282] In addition, the present invention includes all pharmaceutically acceptable antibody drug conjugates of formula (I) or salts thereof, and compounds of formula (II) or salts thereof, which are labeled with one or more radioactive or non-radioactive isotopes. Examples of suitable isotopes used in the isotope labeling of the compounds of the present invention include hydrogen ( 2 H and 3 H, etc.), carbon ( 11 C. 13 C and 14 C, etc.), nitrogen ( 13 N and 15 N, etc.), oxygen ( 15 O. 17 O and 18 O, etc.), fluorine ( 18 F, etc.), chlorine ( 36 Cl, etc.), iodine ( 123 I and 125 I, etc.), phosphorus ( 32 P, etc.), sulfur ( 35 S, etc. The compounds of the present invention labeled with isotopes can be used for research on drug and / or matrix tissue distribution, etc. For example, tritium ( 3 H), carbon 14 ( 14 Radioactive isotopes such as deuterium (C) can be used for this purpose due to the ease of labeling and the simplicity of detection. 2 H), which can sometimes be therapeutically advantageous due to increased metabolic stability (e.g., increased half-life in vivo, reduced dosage requirements, reduced drug interactions). Substitution with positron-emitting isotopes ( 11 C. 18 F. 15 O and 13N, etc.) can be used in positron emission tomography (PET) experiments to test substrate receptor occupancy. The isotope-labeled compounds of the present invention can usually be prepared by conventional methods known to those skilled in the art, or by using an isotope-labeled appropriate reagent instead of an unlabeled reagent and using the same preparation method as in the Examples or Preparation Examples.

[0283] (Aspects of the present invention [3])

[0284] One embodiment of the antibody according to [3] of the present invention is as follows.

[0285] (AB1) an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 2, and a light chain variable region consisting of amino acids 1 to 108 of SEQ ID NO: 4; or an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 6, and a light chain variable region consisting of amino acids 1 to 108 of SEQ ID NO: 8.

[0286] (AB2) An anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 and a light chain consisting of the amino acid sequence of SEQ ID NO: 4; or an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 6 and a light chain consisting of the amino acid sequence of SEQ ID NO: 8.

[0287] (AB3) The anti-CLDN6 antibody described in (AB2) which has been post-translationally modified.

[0288] (AB4) An anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 which is post-translationally modified, and a light chain consisting of the amino acid sequence of SEQ ID NO: 4; or an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 6 which is post-translationally modified, and a light chain consisting of the amino acid sequence of SEQ ID NO: 8.

[0289] (AB5) An antibody-drug conjugate comprising the anti-CLDN6 antibody or antigen-binding fragment thereof according to any one of (AB1) to (AB4).

[0290] In the antibody-drug conjugate comprising the anti-CLDN6 antibody of the present invention described in (AB5), in addition to the TLR7 / 8 dual agonist compound of the present invention, drugs and linkers known in the art may also be used.

[0291] (Manufacturing method)

[0292] The antibody drug complex of formula (I) and its salt and the compound of formula (II) or its salt can utilize the characteristics based on its basic structure or substituent type, adopt various known synthesis methods to manufacture.At this time, according to the type of functional group, in the stage from raw material to intermediate, the functional group is replaced with a suitable protecting group (a group that can be easily converted into the functional group), which is sometimes effective in manufacturing technology.As such a protecting group, for example, Wuts (PGM Wuts) and Greene (TW Greene) can be cited, and the protecting groups recorded in "Greene's Protective Groups in Organic Synthesis (4th edition, 2006)" etc., as long as they are appropriately selected and used according to their reaction conditions.In this method, after the protecting group is introduced and reacted, the protecting group is removed as needed, and thus the desired compound can be obtained.

[0293] Representative methods for preparing the antibody-drug complex of formula (I) or its salt, and the compound of formula (II) or its salt are described below. Each method can also be performed with reference to the references attached to the description. It should be noted that the preparation method of the present invention is not limited to the examples shown below.

[0294] (The first method for preparing antibody-drug complex)

[0295] The antibody-drug conjugate included in the present invention can be produced by the following production method.

[0296] [Chemical formula 16]

[0297]

[0298] (Wherein, Ab represents an antibody or an antigen-binding fragment thereof.)

[0299] The antibody-drug complex (I) of the present invention can be produced by the Michael addition reaction of the antibody or its antigen-binding fragment (1) and the compound (II-L) of the present invention. Specifically, the antibody-drug complex (I) of the present invention can be produced as follows: after reducing the disulfide bonds between the L chain and H chain and / or the disulfide bonds in the hinge part of the antibody or its antigen-binding fragment (1), the generated thiol part is subjected to Michael addition to the compound (II-L). Generally, the antibody is incubated in a neutral to acidic buffer in the presence of a reducing agent at 20 to 37°C for 1 to 24 hours, and then the compound (II-L) is added for incubation. As a solution, a solution containing a phosphate buffer can be used. As a phosphate buffer, for example, disodium hydrogen phosphate can be used. For example, a PBS6.2 / EDTA solution can be used. As a reducing agent, for example, TCEP can be used.

[0300] (First method for preparing drug-linker complex)

[0301] The drug-linker conjugate can be produced by the following method: The drug-linker conjugate (ie, a compound in which M in formula (II) is a group represented by formula (d)) is used to produce the antibody-drug conjugate of formula (I).

[0302] [Chemical formula 17]

[0303]

[0304] (Where R 41 YesR 40 Where M is a group of H. 41’ YesR 40 A divalent group that replaces H of M. -(C=O)-L 1 Indicates L D -(C=O)lower alkylene-, -(C=O)NHlower alkylene-, -(C=O)lower alkylene-(OCH2CH2) m -NH-(C=O)-lower alkylene-, or -(C=O)-cyclohexanediyl-lower alkylene-. The same shall apply to the following.)

[0305] The compound (II-La) of the present invention is a compound (II) of the present invention wherein L is -C(=O)L 1 The compound (II-La) of the present invention can be produced by an amidation reaction of the compound (II-1) and the compound (La).

[0306] In this reaction, the compound (II-1) of the present invention and the compound (La) are used in equal amounts or in excess, and the mixture thereof is stirred in the presence of a condensing agent in a solvent inert to the reaction, under cooling or heating, in one embodiment, at -20°C to 60°C, usually for 0.1 hour to 5 days. Examples of the solvent include: aromatic hydrocarbons such as benzene, toluene, and xylene; halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, and chloroform; ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; DMF, DMSO, EtOAc, MeCN, or water, and mixtures thereof. Examples of the condensing agent include HATU, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, dicyclohexylcarbodiimide, 1,1'-carbonyldiimidazole, diphenylphosphoryl azide, and phosphoryl chloride, but are not limited thereto. The use of an additive (e.g., 1-hydroxybenzotriazole) is sometimes beneficial for the reaction. Conducting the reaction in the presence of an organic base such as triethylamine, DIPEA, or N-methylmorpholine; or an inorganic base such as potassium carbonate, sodium carbonate, or potassium hydroxide is sometimes beneficial for the reaction to proceed smoothly.

[0307] Alternatively, a method may be employed in which carboxylic acid (La) is converted into a reactive derivative and then reacted with an amine of compound (II-1). Examples of reactive derivatives of carboxylic acids include acid halides obtained by reaction with a halogenating agent such as phosphorus oxychloride or thionyl chloride, mixed acid anhydrides obtained by reaction with isobutyl chloroformate, and active esters obtained by condensation with 1-hydroxybenzotriazole. The reaction of these reactive derivatives with compound (II-1) can be carried out in a solvent inert to the reaction such as a halogenated hydrocarbon, an aromatic hydrocarbon, or an ether, under cooling or heating, preferably at -20°C to 60°C.

[0308] [Reference] SR Sandler and W. Karo, "Organic Functional Group Preparations", 2nd edition, volume 1, Academic Press Inc., 1991. The Chemical Society of Japan, ed., "Experimental Chemistry Lectures (5th edition)", volume 16 (2005) (Maruzen).

[0309] (Drug-Linker Complex Preparation Method 2)

[0310] [Chemical formula 18]

[0311]

[0312] (Where L 2 represents a lower alkylene group after removing -CH2- from L. The same shall apply hereinafter.)

[0313] The compound (II-Lb) of the present invention is a compound (II) of the present invention wherein L is -CH2-L 2 The compound of the present invention (II-Lb) can be produced by a reductive amination reaction of the compound of the present invention (II-1) and the compound (Lb).

[0314] In this reaction, compound (II-1) and compound (Lb) are used in equal amounts or in excess, and a mixture thereof is stirred in a solvent inert to the reaction at -45°C to reflux, preferably at 0°C to room temperature, in the presence of a reducing agent, for usually 0.1 hour to 5 days. Examples of the solvent are not particularly limited, and examples thereof include alcohols such as methanol and ethanol; ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; and mixtures thereof. Examples of the reducing agent include sodium cyanoborohydride, sodium triacetoxyborohydride, and sodium borohydride. It is sometimes preferred to carry out the reaction in the presence of a dehydrating agent such as molecular sieves or an acid such as acetic acid, hydrochloric acid, and isopropyl titanate complex. Depending on the reaction, an imine is sometimes generated by the condensation of compound (II-1) and compound (Lb), and can be isolated as a stable intermediate. In this case, compound (II-Lb) can be produced by a reduction reaction of the imine intermediate. In addition, instead of using the reducing agent, the reaction may be carried out in a solvent such as methanol, ethanol, EtOAc, in the presence or absence of an acid such as acetic acid or hydrochloric acid, using a reducing catalyst (e.g., palladium carbon, Raney nickel, etc.). In this case, it is preferred that the reaction be carried out under a hydrogen atmosphere of normal pressure to 50 atmospheres, under cooling or heating.

[0315] [Reference] AR Katritzky and RJK Taylor, "Comprehensive Organic Functional Group Transformations II", Vol. 2, Elsevier Pergamon, 2005. The Chemical Society of Japan, ed., "Experimental Chemistry Lectures (5th Edition)", Vol. 14 (2005) (Maruzen).

[0316] (Third method for preparing drug-linker complex)

[0317] [Chemical formula 19]

[0318]

[0319] (Where R 42 Represents R 40 wherein -CH2- of the group represented by formula (a1), formula (b1) or formula (c1) is removed, and M represents a bond. In ring B, G is N. R 42 , indicating R 42 A divalent group obtained by removing H from NH in the formula (Hal) represents a leaving group. Pr1 represents a protecting group and Pr2 represents H, or Pr1 and Pr2 represent a protecting group as a whole. The same shall apply to the following.)

[0320] The compound (II-Lc) of the present invention can be prepared as follows: Compound (II-Lc-NH2) obtained by deprotecting compound (II-Lc-Pr) prepared by the reaction of compound (2) and compound (Lc-Pr) is subjected to a cyclization reaction. Here, examples of leaving groups include halogen, methanesulfonyloxy, p-toluenesulfonyloxy, etc. Examples of protecting groups Pr1 include tert-butoxycarbonyl, etc., and examples of Pr1 and Pr2 being integrated include phthalimide, etc. In the reaction to obtain compound (II-Lc-Pr), compound (2) and compound (Lc-Pr) are used in equal amounts or one of them is used in excess, and their mixture is stirred in a solvent inert to the reaction or in the absence of a solvent, under cooling or under heating reflux, preferably at 0°C to 80°C, usually for 0.1 hour to 5 days. Examples of solvents include aromatic hydrocarbons such as benzene, toluene, and xylene; ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, and chloroform; DMF, DMSO, EtOAc, MeCN, and mixtures thereof. It is sometimes advantageous to conduct the reaction in the presence of an organic base such as triethylamine, DIPEA, or N-methylmorpholine; or an inorganic base such as cesium carbonate, potassium carbonate, sodium carbonate, or potassium hydroxide in order to facilitate the reaction.

[0321] [Reference] SR Sandler and W. Karo, "Organic Functional Group Preparations", 2nd edition, volume 1, Academic Press Inc., 1991. The Chemical Society of Japan, ed., "Experimental Chemistry Lectures (5th edition)", volume 14 (2005) (Maruzen).

[0322] The deprotection of compound (II-Lc-Pr) to compound (II-Lc-NH2) can be carried out using commonly used conditions.

[0323] In the reaction of preparing compound (II-Lc) from compound (II-Lc-NH2), N-methoxycarbonylmaleimide is added to compound (II-Lc-NH2) in a solvent in the presence of a base, and the mixture is stirred at room temperature for 0.5 to 4 hours. Examples of solvents include aromatic hydrocarbons such as benzene, toluene, and xylene; ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; and DMF. As a base, an aqueous sodium bicarbonate solution can be used.

[0324] (Drug Preparation Method No. 1)

[0325] The drug (ie, the compound wherein M is H in formula (II)) can be produced by the following production method.

[0326] [Chemical formula 20]

[0327]

[0328] The compound (II) of the present invention can be prepared by reacting the compound (3) with the compound (4). In the reaction, the compound (3) and the compound (4) are used in equal amounts or in excess, and the mixture thereof is stirred in a solvent inert to the reaction or in the absence of a solvent, from cooling to heating under reflux, under microwave irradiation, at 0°C to 100°C, usually for 0.1 hour to 5 days. Examples of solvents include: aromatic hydrocarbons such as benzene, toluene, and xylene; ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, and chloroform; DMF, DMSO, EtOAc, MeCN, and mixtures thereof. It is sometimes advantageous to conduct the reaction in the presence of an organic base such as triethylamine, DIPEA, or N-methylmorpholine; or an inorganic base such as cesium carbonate, potassium carbonate, sodium carbonate, or potassium hydroxide in order to facilitate the reaction.

[0329] [Reference] SR Sandler and W. Karo, "Organic Functional Group Preparations", 2nd edition, volume 1, Academic Press Inc., 1991. The Chemical Society of Japan, ed., "Experimental Chemistry Lectures (5th edition)", volume 14 (2005) (Maruzen).

[0330] It should be noted that in the above steps, R 40 Using R with a protecting group 40 The reaction is carried out and finally the protecting group is deprotected.

[0331] (Drug Preparation Method 2)

[0332] [Chemical formula 21]

[0333]

[0334] (Where R 43 Represents R 40 In the formula (a1), formula (b1) or formula (c1), the group after removing -CH2-, G is N, and M is a protecting group or H.)

[0335] The compound (II-1) of the present invention can be produced by subjecting the compound (6) obtained by the oxidation reaction of the compound (5) to a reductive amination reaction with the compound (7).

[0336] In the oxidation reaction of compound (5), compound (5) is treated with an equal amount or excess amount of an oxidizing agent in a solvent inert to the reaction, under cooling or heating, preferably at -20°C to 80°C, for usually 0.1 hour to 3 days. Examples of the solvent used here are not particularly limited, and include: ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, or chloroform; aromatic hydrocarbons such as benzene, toluene, and xylene; DMF, DMSO, EtOAc, MeCN, water, or a mixture thereof. As the oxidizing agent, for example, hydrogen peroxide, cumene hydroperoxide, peracetic acid, perbenzoic acid, m-chloroperbenzoic acid, potassium persulfate (Oxone), activated manganese dioxide, chromic acid, potassium permanganate, and sodium periodate are preferably used. In this reaction, oxidation using DMSO such as Swern oxidation or oxidation using a Dess-Martin reagent is preferably used.

[0337] [References] BMTrost, "Comprehensive Organic Synthesis", Vol. 7, 1991. M. Hudlicky, "Oxidation in Organic Chemistry (ACS Monograph: 186)", ACS, 1990. The Chemical Society of Japan, "Experimental Chemistry Lectures (5th Edition)", Vol. 17 (2005) (Maruzen).

[0338] In the reductive amination reaction of preparing the compound (II-1) of the present invention from the compound (6) and the compound (7), the same method as the method used in preparing the compound (II-Lb) of the present invention from the compound (II-1) and the compound (Lb) described in (Drug-linker complex preparation method 2) can be used.

[0339] It should be noted that in the above steps, the HR of compound (7) 43 In, R 43 The compound may have a protecting group, in which case the compound (II-1) of the present invention can be produced by finally performing deprotection. The deprotection can be carried out under commonly used conditions.

[0340] (Drug Preparation Method 3)

[0341] [Chemical formula 22]

[0342]

[0343] (Ring B' is a 4- to 6-membered cyclic amine composed of multiple Cs, 1 N and 1 G, G is C, and has one unsaturated bond. Ring B" is a saturated ring after one unsaturated bond of ring B' is reduced, and G is CH. Pr represents a protecting group.)

[0344] The same applies below.)

[0345] The compound (II-2) of the present invention can be produced by subjecting the compound (2) obtained from the compound (5) to a Suzuki-Miyaura coupling reaction with the compound (8), followed by hydrogenation and deprotection.

[0346] Compound (2) can be prepared by halogenating compound (5). The halogenation reaction can be carried out under conditions commonly used for halogenation. A halogenating agent is added to a solution of dichloromethane or the like under ice cooling, and the mixture is stirred at room temperature for 1 to 4 hours. As the halogenating agent, thionyl chloride or the like can be used.

[0347] In the Suzuki-Miyaura coupling reaction of compound (2) and compound (8), the boronic acid group, i.e., B(OH)2 group, of compound (8) may be a borate ester group, a boronic acid pinacol ester group, a triol borate ester group, or a trifluoroborate group. In this reaction, compound (2) and compound (8) are used in equal amounts or one of them is used in excess, and their mixture is stirred in a solvent inert to the reaction in the presence of a base and a palladium catalyst at room temperature to heating reflux, preferably at 20°C to 140°C, and usually for 0.1 hour to 5 days. Examples of solvents include halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, and chloroform; aromatic hydrocarbons such as benzene, toluene, and xylene; ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; alcohols such as MeOH, EtOH, isopropanol, butanol, and amyl alcohol; DMF, DMSO, MeCN, 1,3-dimethylimidazolin-2-one, water, and mixtures thereof. Examples of bases include inorganic bases such as tripotassium phosphate, sodium carbonate, potassium carbonate, sodium hydroxide, and barium hydroxide. Examples of the palladium catalyst include tetrakis(triphenylphosphine)palladium, bis(triphenylphosphine)palladium(II) dichloride, [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride / dichloromethane adduct, (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one / palladium (3:2), (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate, palladium(II) acetate, etc. It is sometimes advantageous to conduct the reaction in the presence of a ligand such as dicyclohexyl(2',6'-dimethoxybiphenyl-2-yl)phosphine, dicyclohexyl(2',6'-diisopropoxy-[1,1'-biphenyl]-2-yl)phosphine, or 1,1'-bis(diphenylphosphino)ferrocene in order to facilitate the reaction. In addition, heating the mixture by microwave irradiation is sometimes advantageous in making the reaction proceed smoothly.

[0348] [References] J. Am. Chem. Soc., 2005, 127, p. 4685-4696, Org. Lett. 2011, 13, p. 3948-3951; Org. Lett. 2012, 14, p. 1278-1281.

[0349] The hydrogenation reaction of the compound obtained by the reaction of the compound (2) in step 1) is carried out by stirring the compound in a hydrogen atmosphere in a solvent inert to the reaction in the presence of a metal catalyst for usually 1 hour to 5 days. The reaction is usually carried out under cooling or heating, preferably at room temperature. Examples of the solvent are not particularly limited, and include alcohols such as methanol, ethanol, and 2-propanol; ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; water, EtOAc, DMF, DMSO, and mixtures thereof. As the metal catalyst, palladium catalysts such as palladium carbon, palladium black, and palladium hydroxide; platinum catalysts such as platinum plate and platinum oxide; nickel catalysts such as reduced nickel and Raney nickel; rhodium catalysts such as tetrakistriphenylphosphine rhodium chloride; iron catalysts such as reduced iron, etc. may be preferably used. Formic acid or ammonium formate in an equal amount to an excess amount relative to the compound may be used as a hydrogen source instead of hydrogen gas.

[0350] [Reference] M. Hudlicky, “Reductions in Organic Chemistry, 2nd ed (ACS Monograph: 188)”, ACS, 1996. The Chemical Society of Japan, ed., “Experimental Chemistry Lectures (5th ed.),” Vol. 19 (2005) (Maruzen).

[0351] The compound (II-2) of the present invention can be produced by deprotecting the protecting group Pr. The protecting group Pr of the amine can be deprotected under conditions generally used for deprotection.

[0352] (Raw material synthesis 1)

[0353] [Chemical formula 23]

[0354]

[0355] (In the formula, Lv represents a leaving group.)

[0356] Compound (3) can be prepared from compound (9) and compound (10). Examples of the leaving group include halogen, methanesulfonyloxy, p-toluenesulfonyloxy, etc. The same conditions as those for the alkylation reaction in preparing compound (II-Lc-Pr) from compound (2) and compound (Lc-Pr) described in (Drug-Linker Complex Preparation Method 3) can be used.

[0357] (Raw material synthesis 2)

[0358] [Chemical formula 24]

[0359]

[0360] (In the formula, Pr3 represents a protecting group.)

[0361] Compound (5) can be produced by subjecting Compound (11) produced from Compound (3-1) and Compound (4) to a reduction reaction.

[0362] Compound (3-1) can be prepared in the same manner as in the above-mentioned starting material synthesis 1. In this case, R of compound (10) is used. 40 The substitution reaction of compound (3-1) and compound (4) to compound (11) can be carried out under the same conditions as those described in (Drug Preparation Method 1) for preparing compound (II) of the present invention from compound (3) and compound (4).

[0363] In the reduction reaction from compound (11) to compound (5), the reaction conditions for reducing an ester to an alcohol can generally be used. Compound (11) is treated with an equal amount or an excess amount of a reducing agent in a solvent inert to the reaction, under cooling or heating, preferably at -20°C to 80°C, for usually 0.1 hour to 3 days. Examples of the solvent used here are not particularly limited, and include ethers such as diethyl ether, THF, 1,4-dioxane, and dimethoxyethane; alcohols such as methanol, ethanol, and 2-propanol; aromatic hydrocarbons such as benzene, toluene, and xylene; DMF, DMSO, EtOAc, and mixtures thereof. As the reducing agent, a hydride reducing agent such as LAH, sodium borohydride, or diisobutylaluminum hydride; a metal reducing agent such as sodium, zinc, and iron are preferably used. In addition, the reducing agents described in the following literature can also be used appropriately.

[0364] [Reference] TJ Donohoe, “Oxidation and Reduction in Organic Synthesis (Oxford Chemistry Primers 6)”, Oxford Science Publications, 2000. The Chemical Society of Japan, ed., “Experimental Chemistry Lectures (5th Edition)”, Vol. 14 (2005) (Maruzen).

[0365] (Raw material synthesis 3)

[0366] [Chemical formula 25]

[0367]

[0368] (wherein, Pr4 represents a protecting group, Lv' represents a leaving group, and R 44 Represents R 40 In the formula (a1), formula (b1) or formula (c1), the group after removing -CH2-, G is N. )

[0369] Compound (10) can be prepared by subjecting Compound (14) prepared from Compound (12) and Compound (13) to a reduction reaction, and then replacing the hydroxyl group with a leaving group. Examples of the leaving group include halogen, methanesulfonyloxy, p-toluenesulfonyloxy, and the like.

[0370] Compound (14) can be produced in the same manner as in the starting material synthesis 1. Compound (15) can be produced in the same manner as in the production of compound (11) to (5) in the starting material synthesis 1.

[0371] The antibody drug conjugate of formula (I) or its salt, and the compound of formula (II) or its salt are separated and purified in the form of free compounds, salts, hydrates, solvates or polymorphic substances. The antibody drug conjugate of formula (I) or its salt, and the compound of formula (II) or its salt can also be prepared by performing conventional salt-forming reactions. Separation and purification are performed by applying extraction, fractional crystallization, various separation chromatography (silica gel chromatography, gel filtration, affinity chromatography, etc.) conventional operations.

[0372] Various isomers can be produced by selecting appropriate raw material compounds, or can be separated by utilizing the differences in physicochemical properties between isomers. For example, optical isomers can be obtained by general optical resolution methods of racemates (e.g., fractional crystallization of diastereoisomer salts with optically active bases or acids, chromatography using chiral columns, etc.), or can be produced from appropriate optically active raw material compounds.

[0373] The pharmacological activities of the antibody-drug conjugate of formula (I) or a salt thereof, the compound of formula (II) or a salt thereof, and the anti-CLDN6 antibody used in the antibody-drug conjugate of formula (I) were confirmed by the following test.

[0374] (Abbreviation)

[0375] Ex: Example (ExL1A1, ExL1A2, ExL5A1 and ExL7A1 are example antibody-drug conjugates. Among them, ExL1-Isotype, ExL5-Isotype and ExL7-Isotype and the examples with -Isotope are antibody-drug conjugates as comparative examples). Ab0: An antibody composed of the heavy chain aCLDN6-VH-CH1-CH2-CH3 and the light chain aCLDN6-VL-CL described in International Publication No. 2022 / 058298, Ab1: An antibody comprising a heavy chain composed of the amino acid sequence of sequence number 2 and a light chain composed of the amino acid sequence of sequence number 4, Ab2: An antibody comprising a heavy chain composed of the amino acid sequence of sequence number 6 and a light chain composed of the amino acid sequence of sequence number 8.

[0376] Experimental Example 1: TLR7 / 8 reporter analysis

[0377] (1. Acquisition of cell lines)

[0378] HEK293 cells (ATCC, #CRL-1573) were transfected with an expression vector carrying a base sequence encoding a TLR (human TLR7, human TLR8 or mouse TLR7) protein and a Nanoluc PEST luciferase reporter vector carrying an NFκB response element sequence (Genbank: JQ513377 bases 33 to 84). Then, the transfected cells were cultured in DMEM (Sigma-Aldrich, #D5796-500mL) containing Geneticin (Gibco, #10131-027) with a final concentration of 800 μg / mL, Hygromycin B (Invitrogen, #10687010) with a final concentration of 500 μg / mL, penicillin-streptomycin (Gibco, #15070-063) with a final concentration of 1%, and 10% fetal bovine serum (FBS) (HyClone, #SH30084.03) to obtain a stably expressing cell line.

[0379] (2. In vitro low molecular screening)

[0380] DMEM containing 10% FBS was added to the reporter cells to prepare 1×10 5 Cells / mL suspension was added to each well of a 384-well collagen I (White Collagen I) coated plate (hereinafter referred to as "384-well plate", Corning, #356703) at 20 μL. Incubate overnight (18 hours) at 37°C and 5% CO2 to allow the cells to adhere to the plate. The test sample was dissolved in DMSO, diluted with DMEM medium containing 10% FBS to a concentration 5 times the final concentration, and added to the 384-well plate at 5 μL / well. Further, after incubation at 37°C and 5% CO2 for 7 hours or 24 hours, the plate was removed and allowed to stand at room temperature for 10 minutes. Then, the chemiluminescence was measured using the Nano-Glo luciferase assay system (Promega, #N1120) and the EnVision 2104 (PerkinElmer) plate reader. A dose-response curve was prepared for each test sample, and the EC was calculated as the concentration that obtained 50% of the maximum signal. 50 The EC values ​​of each compound 50 The values ​​are shown in the following table. In the table, hTLR7 indicates human TLR7, hTLR8 indicates human TLR8, and mTLR7 indicates mouse TLR7. Asterisks (*) indicate results of 7-hour incubation, and the others indicate results of 24-hour incubation.

[0381] [Table 2]

[0382]

[0383] As a result of the above test, it was confirmed that some of the compounds of formula (II) or salts thereof shown in the above table have TLR7 / 8 dual agonist action.

[0384] Test Example 2: Evaluation of Binding Activity to Human CLDN6 Using Flow Cytometry

[0385] The human gastric cancer cell line NUGC-3 (JCRB cell bank) expressing CLDN6 was prepared into 1×10 6 The concentration of cells / mL was inoculated into a 96-well V-bottom microplate at 100 μL / well, and the supernatant was removed after centrifugation. Ab0 is an antibody composed of heavy chain aCLDN6-VH-CH1-CH2-CH3 and light chain aCLDN6-VL-CL described in International Publication No. 2022 / 058298. Ab0, Ab1 and Ab2 were prepared to 1 ng / mL to 25 μg / mL using PBS containing 5% FBS (prepare a 12-point dilution series starting from 25 μg / mL and diluted 2.5 times to 1 ng / mL), add 100 μL / well each, and let stand at 4°C for 1 hour. PE-F(ab')2 goat anti-human IgG (JACKSON, #109-116-098) was diluted 100 times with PBS containing 5% FBS, and the Zombie NIR FixableViability Kit (Biolegend, #423106) was further diluted 200 times with the diluent to prepare a secondary antibody solution. After washing twice with PBS containing 5% FBS, 100 μL / well of the prepared secondary antibody solution was added and allowed to stand at 4°C for 1 hour. After washing twice with PBS containing 5% FBS, the cells were resuspended in PBS containing 5% FBS and detected using CytoFlex S (Beckman Coulter, Inc.). Data analysis was performed using Flowjo (Treestar). A histogram of the PE (phycoerythrin) fluorescence intensity of the living cell portion was prepared, and the geometric mean fluorescence intensity (gMFI) was calculated. Graphing using gMFI values ​​was performed using GraphPad Prism (GraphPad Software). The results of the above test confirmed that Ab0, Ab1, and Ab2 bind to human CLDN6. Figure 1The binding activities of Ab0, Ab1, and Ab2 to human CLDN6 are shown.

[0386] Test Example 3: Evaluation of TNF-α and INF-γ production

[0387] (1. Preparation of effector cells)

[0388] Human peripheral blood mononuclear cells (PBMCs) collected from healthy subjects using Ficoll-Paque PLUS (GE Healthcare Bioscience) according to conventional methods were suspended in RPMI1640 Medium (Thermo Fisher Scientific, #11875-093) containing 10% FBS, and the number of viable cells was determined by trypan blue dye exclusion test, and then 1×10 7 cells / mL were used as effector cells.

[0389] (2. Recovery of culture supernatant)

[0390] NUGC-3 was prepared in RPMI1640 medium containing 10% FBS at a concentration of 7.5×10 6 Cells / mL, and 25μL / well of each were inoculated in a 96-well microplate. PBMC prepared in the above 1 were inoculated at 50μL / well, and ExL1A1 and ExL1A2 were prepared to a final concentration of 5.5pM to 0.97μM in RPMI1640 Medium containing 10% FBS at 25μL / well (prepare a 12-point dilution series starting from 0.97μM and diluted 3-fold to 5.5pM), and cultured at 37°C, 5% CO2 for 24 hours, and the supernatant was recovered.

[0391] (3. Determination of TNF-α and INF-γ production)

[0392] The supernatant recovered in 2 above was used to measure the production of TNF-α and IFN-γ according to conventional methods using TNFα (human) AlphaLISA kit (PerkinElmer, #AL208C) and IFN-γ (human) AlphaLISA kit (Perkin Elmer, #AL217F). The EnVision 2104 plate reader was used for measurement. GraphPad Prism (GraphPad Software) was used for plotting.

[0393] Figure 2 The graphs show the results of measuring the cytokine levels of ExL1A1 and ExL1A2.

[0394] As a result of the above-mentioned tests, cytokine production was confirmed in some of the antibody-drug conjugates of formula (I) or salts thereof shown in the above table.

[0395] Experimental Example 4: In vivo activity evaluation in a xenograft model

[0396] 5×10 6 NUGC-3 cells were suspended in 100 μL of Corning Matrigel basement membrane matrix without phenol red (Corning, #356237) and transplanted subcutaneously into the flank of SCID mice (CB17 / Icr-Prkdc scid / CrlCrlj, purchased from Charles River Laboratories Japan, Inc.) at 100 μL / head. 7 days and 14 days after transplantation, PBS or the test antibody or the test antibody-drug complex was administered to the tail vein of the tumor-bearing mice at 3 mg / kg (n=6 or 9). As the test antibodies, Ab0 and Ab1 were used; as the test antibody-drug complex, ExL1A1, ExL5A1, ExL7A1, ExL1-Isotype, ExL5-Isotype, and ExL7-Isotype were used. ExL1-Isotype is an antibody that uses an anti-KLH antibody (International Publication No. 2013 / 094723) instead of Ab1 of ExL1A1. ExL5-Isotype and ExL7-Isotype were prepared in the same manner as ExL1-Isotype. The long and short axes of the transplanted tumors were measured twice a week using an electronic digital caliper (manufactured by Mitutoyo Corporation), and the tumor volume was calculated using the following calculation formula.

[0397] Tumor volume (mm 3 )=1 / 2×short diameter(mm)×short diameter(mm)×long diameter(mm)

[0398] The results of the above tests showed that no antitumor effect was confirmed in Ab0, Ab1, ExL1-Isotype, ExL5-Isotype, and ExL7-Isotype, but a significant antitumor effect was observed in ExL1A1, ExL5A1, and ExL7A1 ( Figure 3). According to the results, some of the anti-CLDN6 antibody-drug complexes containing TLR7 / 8 dual agonist compounds have confirmed anti-tumor effects on cancers expressing CLDN6. The antibody-drug complexes of the present invention containing TLR7 / 8 dual agonist compounds can be expected to be used for the prevention or treatment of such cancers. In addition, the anti-TAA antibody-drug complexes of the present invention containing TLR7 / 8 dual agonist compounds can be expected to be used for the prevention or treatment of cancers expressing the TAA.

[0399] Test Example 5: Evaluation of INF-γ production when adding TLR8 and TLR7 / 8 dual inhibitors

[0400] 2×10 cells were prepared by using RPMI1640 medium containing 10% FBS and introducing GFP into the human ovarian cancer cell line OVCAR-3 (ATCC, #HTB-161) expressing CLDN6. 6 cells / mL, 25 μL / well were inoculated in each well of a 96-well microplate, and 1.25×10 7 40 μL / well of PBMCs were inoculated at 10 μL / well of each cell / mL. 10 μL / well of TLR8 inhibitor CU-CPT9a (InvivoGen, #inh-cc9a) or TLR7 / 8 dual inhibitor Enpatoran (MedChemExpress, #HY-134581) prepared with RPMI1640 Medium containing 10% FBS or RPMI1640 Medium containing 10% FBS was added, and cultured at 37°C, 5% CO2 for 3 hours. Then, 25 μL / well of ExL1A1 prepared with RPMI1640 Medium containing 10% FBS to a final concentration of 23 pM to 1.8 μM was added (8-point dilution series starting from 1.8 μM and diluted 5-fold to 23 pM were prepared), and cultured at 37°C, 5% CO2 for 70 hours, and the supernatant was collected. The recovered supernatant was used to measure the amount of INF-γ produced by the same method as in the above Test Example 3 (3. Measurement of TNF-α and INF-γ production). Graphs were drawn using GraphPad Prism (GraphPad Software).

[0401] Figure 4 The results are the results of measuring the amount of INF-γ produced by ExL1A1 when the culture medium was added or when CU-CPT9a or Enpatoran was added.

[0402] The above test results confirm that when a TLR8 inhibitor is added to the antibody-drug complex of the present invention, the production of cytokines is partially inhibited, and when a TLR7 / 8 dual inhibitor is added, the production of cytokines is completely inhibited. Therefore, in addition to the effect of the TLR7 agonist, the amount of cytokine production in the drug-antibody complex of the present invention is also increased due to the contribution of the TLR8 agonist effect.

[0403] A pharmaceutical composition containing one or more antibody-drug conjugates of formula (I) or salts thereof as active ingredients can be prepared by commonly used methods using excipients commonly used in the art, i.e., pharmaceutical excipients or pharmaceutical carriers.

[0404] Administration may be any form of non-oral administration such as intra-articular, intravenous, intramuscular injections, drip preparations, suppositories, eye drops, eye ointments, transdermal liquid preparations, ointments, transdermal patches, transmucosal liquid preparations, transmucosal patches, inhalants, etc. Injections and drip preparations may be administered by appropriate methods such as intravenous administration, subcutaneous administration, intraperitoneal administration, and intratumor administration.

[0405] Injections for parenteral administration contain sterile aqueous or non-aqueous solutions, suspensions or emulsions. As aqueous solvents, for example, distilled water for injection or physiological saline are included. As non-aqueous solvents, for example, alcohols such as ethanol are included. Such compositions may also contain isotonic agents, preservatives, wetting agents, emulsifiers, dispersants, stabilizers or dissolution aids. They are sterilized, for example, by filtering through a bacteria-retaining filter, adding a bactericide or irradiating. In addition, they may also be made into sterile solid compositions, which are dissolved or suspended in sterile water or a sterile injection solvent before use.

[0406] The external preparations include ointments, plasters, creams, jelly, pastes, sprays, lotions, eye drops, eye ointments, etc. They contain common ointment bases, lotion bases, aqueous or non-aqueous liquid preparations, suspensions, emulsions, etc.

[0407] Transmucosal agents such as inhalants or nasal agents can use solid, liquid or semisolid preparations and are manufactured according to previously known methods. For example, known excipients may be appropriately added, or pH adjusters, preservatives, surfactants, lubricants, stabilizers or viscosity enhancers may be further appropriately added. Appropriate inhalation or blowing devices may be used for administration. For example, known devices such as metered dosing inhalation devices or sprayers may be used to administer the compound alone or as a mixed powder after formulation, or the compound may be combined with a pharmaceutically acceptable carrier to be administered in the form of a solution or suspension. Dry powder inhalers and the like may be used for single or multiple administrations, and dry powders or capsules containing powders may be used. Alternatively, pressurized aerosol sprays using appropriate propellants, such as appropriate gases such as chlorofluorocarbons or carbon dioxide, may also be used.

[0408] Usually, the dosage per day is about 0.001 to 100 mg / kg, preferably 0.1 to 30 mg / kg, and more preferably 0.1 to 10 mg / kg, which is administered once or divided into 2 to 4 times. In the case of intravenous administration, the dosage per day is about 0.0001 to 10 mg / kg, which is administered once a day or divided into multiple times. In addition, as a transmucosal agent, it is about 0.001 to 100 mg / kg, which is administered once a day or divided into multiple times. Considering the symptoms, age, sex, etc., the dosage is appropriately determined according to each situation.

[0409] The pharmaceutical composition of the present invention contains 0.01 to 100% by weight, or in a certain embodiment 0.01 to 50% by weight of an active ingredient, i.e., one or more antibody-drug conjugates of formula (I) or salts thereof, although it varies depending on the administration route, dosage form, administration site, and type of excipients or additives.

[0410] The antibody drug conjugate of formula (I) can be used in combination with various therapeutic agents or preventive agents for diseases for which the antibody drug conjugate of formula (I) is believed to be effective. The combination can be administered simultaneously, or administered continuously, or administered at desired time intervals. The preparations for simultaneous administration can be compound preparations, or can be prepared separately. As the drugs used in combination, therapeutic agents for treating the target disease can be listed.

[0411] Example

[0412] Below, the method for producing the antibody drug conjugate of formula (I) is described in more detail based on the examples. It should be noted that the present invention is not limited to the compounds described in the following examples. In addition, the method for producing the raw material compound is recorded as a production example, and the production of the known compound is recorded as a reference example. In addition, the method for producing the antibody drug conjugate of formula (I) is not limited to the production method of the specific examples shown below, and the antibody drug conjugate of formula (I) can also be produced by a combination of these production methods or a method that is obvious to those skilled in the art.

[0413] In addition, the following abbreviations may be used in this specification, examples, preparation examples and tables.

[0414] Anti-CLDN6 antibody: Anti-Claudin 6 antibody, TLR7 / 8 dual agonist: Dual agonist of Toll-like receptor 7 and Toll-like receptor 8, DAST: N,N-diethylaminosulfur trifluoride, DEAD: diethyl azodicarboxylate, DCM: dichloromethane, DIAD: diisopropyl azodicarboxylate, DIPEA: N,N-diisopropylethylamine, DMF: dimethylformamide, DMP: Dess-Martin periodinane, DMSO: dimethyl sulfoxide, D-PBS: Dulbecco's phosphate buffered saline, EtOAc: ethyl acetate, FA: formic acid, HATU: 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, LAH: hydrogenated Lithium aluminum, MeCN: acetonitrile, MsCl: methanesulfonyl chloride, NAC: N-acetyl-L-cysteine, NMP: N-methyl-2-pyrrolidone, PE: petroleum ether, PBS6.2 / EDTA: phosphate buffer containing ethylenediaminetetraacetic acid (EDTA, 5mM) (prepared to pH 6.2 using 10mM, 1M hydrochloric acid), PdCl2(dppf)_CH2Cl2: [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex, Palau'Chlor: 2-chloro-1,3-bis(methoxycarbonyl)guanidine, prep-HPLC: preparative High Performance Liquid Chromatography Chromatography), SEC: size exclusion chromatography, TBAI: tetrabutylammonium iodide, TEA: triethylamine, TCEP: tri(2-carboxyethyl)phosphine hydrochloride, TFA: trifluoroacetic acid, THF: tetrahydrofuran, MWCO: molecular weight cutoff (e.g., 10000 MWCO means that substances with a molecular weight of 10000 or less can be removed by the filter), nPn: n-pentyl, Me: methyl, Et: ethyl, t-Bu: tert-butyl, DAR: compound to antibody ratio in the antibody drug conjugate (i.e., the average number of compounds bound to one antibody), NUM: Example Number or manufacturing example number ( / HCl indicates that the embodiment or manufacturing example is hydrochloride, / TFA indicates that the embodiment or manufacturing example is trifluoroacetate), PEx: manufacturing example number, Ex: example number, REF: manufacturing method (indicates that the substance is manufactured by the same manufacturing method as the substance with the example number or manufacturing example number recorded in this column. For example, when Ex1 is recorded in REF of PEx2-2, it means that the manufacturing example PEx2-2 is manufactured in the same way as the method recorded in Example Ex1. In addition, when multiple numbers are recorded in REF, it means that the manufacturing is carried out in the same order as the steps recorded.For example, when recorded as PEx4-1 or Ex2 in Ex11, it means that the product is produced in the order of the recorded steps), STR: chemical structural formula, DAT: physicochemical data, ESI+: m / z value in ESI-MS+ ([M+H] unless otherwise specified). + ), NMR signals represent representative signals. s: singlet, d: doublet, t: triplet, q: quartet, m: multiplet, br: broad peak, [α] D 20 : specific rotation at 20°C, c: represents the concentration (g / 100mL) when measuring the specific rotation, and also represents the measurement solvent. It should be noted that in this specification, the naming of compounds sometimes uses naming software such as ACD / Name (registered trademark, Advanced Chemistry Development, Inc.). In addition, for convenience, the concentration mol / L is expressed as M. For example, 1M sodium hydroxide aqueous solution refers to 1 mol / L sodium hydroxide aqueous solution.

[0415] Production Examples and Examples

[0416] (Production of Antibodies)

[0417] Production Example AB-1: Preparation of expression vector for humanized anti-CLDN6 antibody

[0418] The humanized anti-CLDN6 antibody is based on the sequences of the heavy chain variable region and the light chain variable region of the mouse anti-CLDN6 antibody GT512muMAB 64A and GT512muMAB 61D described in International Publication No. 2011 / 057788. According to the method described in the literature (Front Biosci., 2008, Vol. 13, p. 1619-1633), the sequence design for the humanization of the mouse anti-CLDN6 antibody was performed. The model structure was constructed and analyzed using the comprehensive computational chemistry system MOE provided by MOLSIS Inc., and back mutations were introduced into the framework region. The two humanized anti-CLDN6 antibodies produced are referred to as Ab1 and Ab2, respectively. Ab1 is an antibody comprising a heavy chain consisting of the amino acid sequence of sequence number 2 and a light chain consisting of the amino acid sequence of sequence number 4, and Ab2 is an antibody comprising a heavy chain consisting of the amino acid sequence of sequence number 6 and a light chain consisting of the amino acid sequence of sequence number 8. The polynucleotides encoding the designed amino acid sequences of Ab1 and Ab2 were loaded on pcDNA 3.4TOPO vector (ThermoFisher scientific company). The prepared vectors were respectively called Ab1 expression vector and Ab2 expression vector.

[0419] Example AB: Preparation of humanized anti-CLDN6 antibodies

[0420] The Ab1 expression vector or Ab2 expression vector prepared in Preparation Example AB-1 was introduced into ExpiCHO-S cells using the ExpiFectamine CHO Transfection Kit (Thermo Fisher Scientific, #A29129) and cultured for several days. Ab1 and Ab2 were purified from the culture supernatant by affinity purification using MabSelect SuRe pcc (GE Healthcare Bioscience, #27-5438-02).

[0421] (Manufacture of drugs)

[0422] Production Example 1-1

[0423] DIPEA (4.72 mL) was added to a solution of (3S)-3-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (3 g) in DMF (30 mL) at 20° C., followed by stirring at 20° C. for 30 minutes, and then methyl 4-(bromomethyl)-2-methoxybenzoate (3.59 g) was added, followed by stirring at 20° C. for 12 hours. The reaction solution was concentrated under reduced pressure, and the obtained oily compound was purified by silica gel column chromatography (PE:EtOAc=50 / 50-0 / 100) to obtain (3S)-3-(hydroxymethyl)-4-{[3-methoxy-4-(methoxycarbonyl)phenyl]methyl}piperazine-1-carboxylic acid tert-butyl ester (5.12 g) as an oily substance.

[0424] Production Example 1-2

[0425] To a solution of (3S)-3-(hydroxymethyl)-4-{[3-methoxy-4-(methoxycarbonyl)phenyl]methyl}piperazine-1-carboxylic acid tert-butyl ester (4.62 g) in chloroform (40 mL) was added a solution of DAST (9.94 g) in chloroform (10 mL) at 0°C, and the mixture was stirred at 20°C for 3 hours. An aqueous sodium bicarbonate solution was added to adjust the pH to 8, and the mixture was extracted twice with DCM (100 mL). The combined organic layers were concentrated under reduced pressure, and the resulting residue was purified by prep-HPLC (column: Xtimate C18 150×40 mm×10 μm, water (0.225% FA) / MeCN=70 / 30-40 / 60) to obtain (3S)-3-(fluoromethyl)-4-{[3-methoxy-4-(methoxycarbonyl)phenyl]methyl}piperazine-1-carboxylic acid tert-butyl ester (2.01 g) as an oily substance.

[0426] Production Example 1-3

[0427] LAH (192.43 mg) was added to a solution of (3S)-3-(fluoromethyl)-4-{[3-methoxy-4-(methoxycarbonyl)phenyl]methyl}piperazine-1-carboxylic acid tert-butyl ester (2.01 g) in THF (35 mL) at 0° C., and the mixture was stirred at 20° C. for 1 hour. Sodium sulfate decahydrate (0.3 g) was added to the reaction solution, and the mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE / EtOAc=50 / 50-20 / 80) to obtain (3S)-3-(fluoromethyl)-4-{[4-(hydroxymethyl)-3-methoxyphenyl]methyl}piperazine-1-carboxylic acid tert-butyl ester (1.71 g) as an oily substance.

[0428] Production Example 1-4

[0429] To a solution of tert-butyl (3S)-3-(fluoromethyl)-4-{[4-(hydroxymethyl)-3-methoxyphenyl]methyl}piperazine-1-carboxylate (1.71 g) in DCM (35 mL) was added DIPEA (1.62 mL) at 0° C., followed by stirring at 20° C. for 30 minutes, and then MsCl (956.97 mg) was added at 0° C., followed by stirring at 20° C. for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain tert-butyl (3S)-4-{[4-(chloromethyl)-3-methoxyphenyl]methyl}-3-(fluoromethyl)piperazine-1-carboxylate (1.8 g) as an oily crude product.

[0430] Production Example 1-5

[0431] Cesium carbonate (2.90 g) was added to a DMF (20 mL) solution of 4-chloro-5H-pyrrolo[3,2-d]pyrimidin-2-amine (0.5 g) at 20°C, stirred at the same temperature for 30 minutes, then (3S)-tert-butyl 4-{[4-(chloromethyl)-3-methoxyphenyl]methyl}-3-(fluoromethyl)piperazine-1-carboxylate (1.03 g) was added, and stirred at the same temperature for 12 hours. The reaction mixture was filtered, water (100 mL) was added to the filtrate, and then extracted three times with ethyl acetate (150 mL). The combined organic layers were concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (PE:EtOAc=0:100) to obtain (3S)-tert-butyl 4-({4-[(2-amino-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxyphenyl}methyl)-3-(fluoromethyl)piperazine-1-carboxylate (1.03 g) as a solid.

[0432] Manufacturing Example 1-6

[0433] To a solution of (3S)-tert-butyl 4-({4-[(2-amino-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxyphenyl}methyl)-3-(fluoromethyl)piperazine-1-carboxylate (0.93 g) in DMSO (20 mL) were added DIPEA (936.34 μL) and pentane-1-amine (312.37 mg) at 20° C., and the mixture was stirred at 80° C. for 12 hours. Water (50 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (100 mL). The combined organic layers were concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (DCM / methanol=10 / 1) to obtain (3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazine-1-carboxylic acid tert-butyl ester (0.9 g) as a solid.

[0434] Example 1

[0435] HCl / 1,4-dioxane (4M, 12 mL) was added to a solution of (3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazine-1-carboxylic acid tert-butyl ester (0.9 g) in DCM (5 mL) at 20°C, and the mixture was stirred at the same temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by prep-HPLC (column: Xtimate C18 150×40 mm×10 μm, water (0.05% NH3 / H2O) / MeCN=90 / 10-40 / 60) to obtain 5-[(4-{[(2S)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N-(3S)-4 ... 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (474 ​​mg).

[0436] Production Example 2-1

[0437] DIPEA (1.61 mL) was added to a solution of (2R)-2-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (1.00 g) in DMF (15 mL) at 20° C., and the mixture was stirred at the same temperature for 30 minutes. Then, methyl 4-(bromomethyl)-2-methoxybenzoate (1.2 g) was added, and the mixture was stirred at 20° C. for 12 hours. The reaction mixture was concentrated under reduced pressure, and the obtained oil was purified by silica gel column chromatography (PE:EtOAc=50 / 50-0 / 100) to obtain (2R)-2-(hydroxymethyl)-4-{[3-methoxy-4-(methoxycarbonyl)phenyl]methyl}piperazine-1-carboxylic acid tert-butyl ester (1.83 g) as an oil.

[0438] Production Example 2-2

[0439] To a solution of (2R)-2-(hydroxymethyl)-4-{[3-methoxy-4-(methoxycarbonyl)phenyl]methyl}piperazine-1-carboxylic acid tert-butyl ester (1.83 g) in DCM (3 mL) was added HCl / MeOH (4M, 30 mL) at 0°C and stirred at 20°C for 12 hours. The reaction mixture was concentrated under reduced pressure, an aqueous sodium bicarbonate solution (150 mL) was added, and then extracted three times with DCM (150 mL). The combined organic layers were concentrated under reduced pressure to obtain methyl 4-{[(3R)-3-(hydroxymethyl)piperazin-1-yl]methyl}-2-methoxybenzoate (1.22 g) as an oil.

[0440] Production Example 2-3

[0441] 2,4-dimethoxybenzaldehyde (688.75 mg) was added to a DCM (20 mL) solution of methyl 4-{[(3R)-3-(hydroxymethyl)piperazin-1-yl]methyl}-2-methoxybenzoate (1.22 g) at 20°C, and the mixture was stirred at 20°C for 4 hours. Then, sodium triacetoxyborohydride (1.14 g) was added, and the mixture was stirred at 20°C for 12 hours. A sodium bicarbonate aqueous solution (50 mL) was added to the reaction mixture, and the mixture was extracted twice with DCM (100 mL). The combined organic layer was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (DCM / methanol=20 / 1) to obtain methyl 4-{[(3R)-4-[(2,4-dimethoxyphenyl)methyl]-3-(hydroxymethyl)piperazin-1-yl]methyl}-2-methoxybenzoate (1.47 g) as an oily substance.

[0442] Production Example 2-4

[0443] Using methyl 4-{[(3R)-4-[(2,4-dimethoxyphenyl)methyl]-3-(hydroxymethyl)piperazin-1-yl]methyl}-2-methoxybenzoate (1.47 g), 5-[(4-{[(3R)-4-[(2,4-dimethoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N-(4-(hydroxymethyl)piperazin-1-yl]methyl}-2-methoxybenzoate was obtained as a solid crude product by the same method as in Preparation Examples 1-2 to 1-6. 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (544.55 mg).

[0444] Example 2

[0445] Under a nitrogen atmosphere, 5-[(4-{[(3R)-4-[(2,4-dimethoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N-[(4-[(2,4-dimethoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]methyl] ...]-5-[(4-[(2,4-dimethoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]methyl]-2-methoxyphenyl]-5-[(fluoromethyl)piperazin-1-yl]methyl]-2-methoxyphenyl]-5-[(fluoromethyl)piperazin-1-yl]methyl]-2-methoxyphenyl]-5-[( 4 TFA (5 mL) was added to a DCM (5 mL) solution of -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (544.55 mg), and the mixture was stirred at 20° C. for 1 hour. The reaction mixture was concentrated under reduced pressure, and the obtained crude product was purified by prep-HPLC (column: Boston Green ODS 150×30 mm×5 μm, water (TFA) / MeCN=94 / 6-54 / 46) to obtain 5-[(4-{[(3R)-3-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N-(4 ... 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (trifluoroacetic acid) salt (240 mg).

[0446] Production Example 3-1

[0447] DIPEA (4.2 mL) and pentane-1-amine (1.6 mL) were added to a solution of {4-[(2-amino-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxyphenyl}methanol (1 g) in DMSO (5 mL), and the mixture was stirred at 100° C. for 2 days. After cooling to room temperature, water and chloroform were added, and the mixture was extracted with chloroform using a phase separator and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol / 28% aqueous ammonia = 100 / 0 / 0-90 / 9 / 1) to obtain (4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methanol (807 mg) as a solid.

[0448] Production Example 3-2

[0449] DMP (90 mg) was added to a mixture of (4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methanol (50 mg) and DCM (2 mL) under ice-cooling, and the mixture was stirred at room temperature for 2 hours. A 10% aqueous sodium sulfite solution, a saturated aqueous sodium bicarbonate solution, and a mixed solvent (chloroform / methanol=5 / 1) were added under ice-cooling, and the mixture was extracted with a mixed solvent (chloroform / methanol=5 / 1) using a phase separator, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol=100 / 0-80 / 20) to obtain 4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxybenzaldehyde (34 mg) as a solid.

[0450] Example 3

[0451] To a mixture of 4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxybenzaldehyde (31 mg), cyclopropylamine (0.012 mL) and TEA (0.024 mL) in DCM (1.2 mL) was added sodium triacetoxyborohydride (36 mg), and the mixture was stirred at room temperature for 2 hours. Cyclopropylamine (0.012 mL) and sodium triacetoxyborohydride (36 mg) were added, and the mixture was stirred at room temperature overnight. Cyclopropylamine (0.012 mL), sodium triacetoxyborohydride (36 mg) and DCM (1.2 mL) were added, and the mixture was stirred at 40°C for 2 hours. After cooling to room temperature, cyclopropylamine (0.012 mL) and sodium triacetoxyborohydride (36 mg) were added, and the mixture was stirred at 40°C for 8 hours. After cooling to room temperature, saturated sodium bicarbonate aqueous solution and chloroform were added, extracted with chloroform using a phase separator, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol / 28% ammonia water=100 / 0 / 0-90 / 9 / 1) to obtain 5-({4-[(cyclopropylamino)methyl]-2-methoxyphenyl}methyl)-N-( ... 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (23 mg).

[0452] Production Example 4-1

[0453] DIPEA (0.162 mL) and pentane-1-thiol (0.094 mL) were added to a solution of {4-[(2-amino-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxyphenyl}methanol (200 mg) in DMSO (2 mL), and the mixture was stirred at 80°C for 2 hours. After cooling to room temperature, DIPEA (0.162 mL) and pentane-1-thiol (0.094 mL) were added, and the mixture was stirred at 100°C overnight. After cooling to room temperature, DIPEA (0.162 mL) and pentane-1-thiol (0.094 mL) were added, and the mixture was stirred at 100°C for 3 days. After cooling to room temperature, water and a mixed solvent (chloroform / methanol) were added, and the mixture was extracted with a mixed solvent (chloroform / methanol) using a phase separator, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol=100 / 0-90 / 10), and then purified by silica gel column chromatography (hexane / ethyl acetate=98 / 2-0 / 100) to obtain (4-{[2-amino-4-(pentylsulfanyl)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methanol (48 mg) as a solid.

[0454] Example 4

[0455] Thionyl chloride (0.066 mL) was added to a DCM (2 mL) solution of (4-{[2-amino-4-(pentylsulfanyl)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methanol (35 mg) under ice cooling, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure. NMP (1 mL), DIPEA (0.078 mL), piperazine (40 mg) and potassium iodide (16 mg) were added to the obtained residue, and the mixture was stirred at 80° C. for 1 hour. After cooling to room temperature, water and a mixed solvent (chloroform / methanol=5 / 1) were added, and the mixture was extracted with a mixed solvent (chloroform / methanol=5 / 1) using a phase separator, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol / 28% aqueous ammonia = 100 / 0 / 0 to 90 / 9 / 1) to obtain 5-({2-methoxy-4-[(piperazin-1-yl)methyl]phenyl}methyl)-4-(pentylsulfanyl)-5H-pyrrolo[3,2-d]pyrimidin-2-amine (33.9 mg) as a solid.

[0456] Production Example 5-1

[0457] DIPEA (0.08 mL) and [(2S)-pyrrolidin-2-yl]methanol (0.035 mL) were added to a MeCN (3 mL) solution of 4-chloro-5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-5H-pyrrolo[3,2-d]pyrimidin-2-amine (200 mg) at room temperature, and the mixture was stirred at the same temperature for 18 hours. The reaction solution was added to water (15 mL) and extracted twice with ethyl acetate (50 mL). Anhydrous sodium sulfate was added to the combined organic layer, filtered, and concentrated under reduced pressure to obtain [(2S)-1-({4-[(2-amino-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxyphenyl}methyl)pyrrolidin-2-yl]methanol (72 mg) as an oily substance.

[0458] Example 5

[0459] [(2S)-1-({4-[(2-amino-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxyphenyl}methyl)pyrrolidin-2-yl]methanol (70 mg), DMSO (1 mL), DIPEA (0.16 mL) and pentane-1-thiol (0.1 mL) were added to a microwave vial and stirred at 100°C for 6 hours under microwave irradiation. The reaction solution was cooled to room temperature, added to water (20 mL), and extracted three times with ethyl acetate (20 mL). The combined organic layer was washed with water and saturated brine, dried over anhydrous MgSO4, and then concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (amino silica gel, ethyl acetate / hexane=50 / 50-100 / 0) to obtain {(2S)-1-[(4-{[2-amino-4-(pentylsulfanyl)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methanol (35 mg) as an oily substance.

[0460] Production Example 6-1

[0461] At room temperature, 5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-N 4 Palau'Chlor (registered trademark) (65 mg) was added to a solution of 1-pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine hydrochloride (0.1 g) in MeCN (3 mL), and the mixture was stirred at the same temperature overnight. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM / methanol = 9 / 1) to obtain 7-chloro-5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-N-(1-(chloromethyl)-2-methoxyphenyl) ... 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine hydrochloride (60 mg).

[0462] Example 6

[0463] To 7-chloro-5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-N 4 DIPEA (0.061 mL) and [(2S)-pyrrolidin-2-yl]methanol (0.024 mL) were added to a DMF (3 mL) solution of pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine hydrochloride (60 mg), and the mixture was stirred at the same temperature overnight. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (amino silica gel, methanol / ethyl acetate = 1 / 99) to obtain {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methanol (33 mg) as a solid.

[0464] Production Example 7-1

[0465] Under ice-cooling, methyl 4-(bromomethyl)-3-methoxybenzoate (1.42 g) was added to a DMF (40 mL) solution of 7-bromo-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-2-amine (2.68 g) and cesium carbonate (1.78 g), and stirred at room temperature for 4 hours. Under ice-cooling, a saturated aqueous ammonium chloride solution (200 mL) was added to the reaction mixture for dilution, and then poured into ice water (600 mL). After filtering out the generated solid, it was washed with ethyl acetate / hexane (1 / 4, 500 mL), thereby obtaining methyl 4-[(2-amino-7-bromo-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxybenzoate (4.44 g) as a solid.

[0466] Production Example 7-2

[0467] To a suspension of LAH (288 mg) in THF (60 mL) was added 4-[(2-amino-7-bromo-4-chloro-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxybenzoic acid methyl ester (4.4 g) under ice-cooling, stirred at the same temperature for 15 minutes, and then sodium sulfate decahydrate (15 g) was added under ice-cooling. The suspension was stirred at room temperature for 5 hours, and the solid was removed by filtration. The obtained filtrate was concentrated under reduced pressure, the crude product was dissolved in DMSO (30 mL), and DIPEA (4 mL) and pentane-1-amine (2 mL) were added. The reaction mixture was stirred at 110°C overnight and then cooled to room temperature. Saturated aqueous ammonium chloride solution (40 mL) and ethyl acetate (100 mL) were added, and the organic layer was washed 3 times with water (100 mL), washed once with saturated brine (100 mL), dried over anhydrous MgSO4, and then concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (amino silica gel, ethyl acetate / hexane=50 / 50) to obtain (4-{[2-amino-7-bromo-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methanol (1.4 g) as a solid.

[0468] Production Example 7-3

[0469] Thionyl chloride (0.13 mL) was added to a solution of (4-{[2-amino-7-bromo-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methanol (0.4 g) in THF (30 mL) at 40°C. The reaction mixture was stirred at the same temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, the resulting residue was diluted with MeCN (20 mL), and then concentrated under reduced pressure again to obtain 7-bromo-5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-N-(4-{[2-amino-7-bromo-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methanol as a solid. 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine hydrochloride (438 mg).

[0470] Example 7

[0471] To a solution of [(2S)-pyrrolidin-2-yl]methanol (0.02 mL), DIPEA (0.1 mL) and TBAI (5 mg) in MeCN (3 mL) was added 7-bromo-5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-N-(2S)-pyrrolidin-2-yl]methanol (0.02 mL), DIPEA (0.1 mL) and TBAI (5 mg) at room temperature. 4-pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine hydrochloride (80 mg) was added, and the reaction mixture was stirred at 50° C. for 16 hours. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (amino silica gel, methanol / chloroform=5 / 95) to obtain {(2S)-1-[(4-{[2-amino-7-bromo-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methanol (65 mg) as a solid.

[0472] Production Example 8-1

[0473] In the same manner as in Production Examples 1-1 and 1-3, tert-butyl 4-{[4-(hydroxymethyl)-3-methoxyphenyl]methyl}piperazine-1-carboxylate (2.6 g) was obtained as an oily substance from methyl 4-(bromomethyl)-2-methoxybenzoate (5 g).

[0474] Production Example 8-2

[0475] Phosphorus oxychloride (892.86 μL) and DIPEA (1.57 mL) were added to a solution of methyl (6-methyl-4-oxo-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)carbamate (1 g) in MeCN (20 mL), and the reaction mixture was stirred at 70° C. for 1 hour. The reaction mixture was cooled to room temperature and injected into water (80 mL) in which sodium acetate (960 mg) was dissolved. The organic layer was concentrated under reduced pressure, and the residue mixture was cooled to 0° C., filtered and washed with MeCN (40 mL). The obtained solid was dried under reduced pressure and then purified by silica gel column chromatography (methanol / DCM=0 / 100-5 / 95) to obtain methyl (4-chloro-6-methyl-5H-pyrrolo[3,2-d]pyrimidin-2-yl)carbamate (300 mg) as a solid.

[0476] Production Example 8-3

[0477] To a mixture of methyl (4-chloro-6-methyl-5H-pyrrolo[3,2-d]pyrimidin-2-yl)carbamate (300 mg), tert-butyl 4-{[4-(hydroxymethyl)-3-methoxyphenyl]methyl}piperazine-1-carboxylate (503.28 mg) and THF (10 mL) were added triphenylphosphine (980.93 mg) and DIAD (484.77 μL), and the mixture was stirred at 25° C. for 1 hour. Water (20 mL) was added to the mixture, and then extracted three times with DCM (10 mL). The combined organic layer was washed three times with saturated brine (5 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (methanol / DCM=0 / 100-5 / 95) to obtain tert-butyl 4-{[4-({4-chloro-2-[(methoxycarbonyl)amino]-6-methyl-5H-pyrrolo[3,2-d]pyrimidin-5-yl}methyl)-3-methoxyphenyl]methyl}piperazine-1-carboxylate (500 mg) as an oily substance.

[0478] Production Example 8-4

[0479] To a solution of tert-butyl 4-{[4-({4-chloro-2-[(methoxycarbonyl)amino]-6-methyl-5H-pyrrolo[3,2-d]pyrimidin-5-yl}methyl)-3-methoxyphenyl]methyl}piperazine-1-carboxylate (500 mg) in 1,4-dioxane (6 mL) was added an aqueous sodium hydroxide solution (2M, 2.73 mL), and the reaction mixture was stirred at 80° C. for 2 hours. Water (10 mL) was added to the reaction mixture, and the mixture was extracted three times with DCM (10 mL). The combined organic layer was washed three times with saturated brine (5 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain tert-butyl 4-({4-[(2-amino-4-chloro-6-methyl-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxyphenyl}methyl)piperazine-1-carboxylate (360 mg) as a solid.

[0480] Example 8

[0481] Using 4-({4-[(2-amino-4-chloro-6-methyl-5H-pyrrolo[3,2-d]pyrimidin-5-yl)methyl]-3-methoxyphenyl}methyl)piperazine-1-carboxylic acid tert-butyl ester (360 mg), 5-({2-methoxy-4-[(piperazin-1-yl)methyl]phenyl}methyl)-6-methyl-N-([(piperazin-1-yl)methyl]phenyl}methyl)-1-carboxylic acid tert-butyl ester was obtained as a solid by the same method as in Preparation Example 1-6 and Example 2. 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine hydrochloride (33.5 mg).

[0482] Production Example 9-1

[0483] Thionyl chloride (0.4 mL) was added to a solution of (4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methanol (200 mg) in DCM (4 mL) under ice cooling, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and a saturated aqueous sodium bicarbonate solution and DCM were added to the obtained residue, extracted with DCM using a phase separator, and concentrated under reduced pressure. To the obtained residue, tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (168 mg), PdCl2(dppf)-CH2Cl2 (24 mg), potassium carbonate (150 mg), 1,4-dioxane (2 mL) and water (0.4 mL) were added, and stirred overnight at 100° C. under an argon atmosphere. After cooling to room temperature, water and chloroform were added, and the mixture was extracted with chloroform using a phase separator, concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (amino silica gel, chloroform / methanol=100 / 0-90 / 10) and silica gel column chromatography (chloroform / methanol=100 / 0-80 / 20) to obtain a solid (109.3 mg). Under an argon atmosphere, 5% platinum-supported carbon (about 50% water content, 38 mg) was added to a solution of the solid (76 mg) in EtOAc (3 mL), and then stirred at room temperature for 2 days under a hydrogen atmosphere. After filtration through Celite (registered trademark), the mixture was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (chloroform / methanol=100 / 0-80 / 20) to obtain 4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]piperidine-1-carboxylic acid tert-butyl ester (65.2 mg) as a solid.

[0484] Example 9

[0485] 4M HCl / dioxane (0.6 mL) was added to a DCM (2 mL) solution of tert-butyl 4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]piperidine-1-carboxylate (62 mg) under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure, and a mixed solvent (chloroform / methanol=5 / 1) and amino silica gel were added to the obtained residue, and the mixture was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (amino silica gel, chloroform / methanol=100 / 0-80 / 20) to obtain 5-({2-methoxy-4-[(piperidin-4-yl)methyl]phenyl}methyl)-N ... 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (41.9 mg).

[0486] Example 10

[0487] A mixture of (3R)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazine-1-carboxylic acid tert-butyl ester (460 mg) and 4M HCl / dioxane (6 mL) in DCM (2 mL) was stirred at 20°C for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC (column: Xtimate C18 150×40 mm×10 μm, water (0.05% NH3 / H2O) / MeCN=90 / 10-40 / 60) to obtain 5-[(4-{[(2R)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N-(3R)-4 ... 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (230 mg).

[0488] (Manufacturing of drug-linker complexes)

[0489] Example L1

[0490] At 20°C, 5-[(4-{[(2S)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionic acid (64.83 mg), HATU (291.49 mg) and DIPEA (0.2 mL) were added to a DMF (5 mL) solution of 1-pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (0.18 g), and the mixture was stirred at the same temperature for 12 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (100 mL). The combined organic layers were concentrated under reduced pressure, and the residue was purified by prep-HPLC (chromatographic column: Phenomenex Gemini-NX C18 75×30 mm×3 μm, water (0.1% TFA)-MeCN=100 / 0-60 / 40) to obtain 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione (trifluoroacetic acid) salt (108.5 mg) as a solid.

[0491] Example L3

[0492] To a mixture of 2,5-dioxopyrrolidin-1-yl 31-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-29-oxo-4,7,10,13,16,19,22,25-octaoxa-28-azatriacontane-1-oic acid 2,5-dioxopyrrolidin-1-yl ester (24 mg), DIPEA (15 μL) and NMP (1 mL) was added 5-[(4-{[(2S)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N ...]- 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (15 mg) was added and stirred at room temperature for 1 hour. The reaction mixture was purified by reverse phase silica gel column chromatography (ODS, water (0.1% TFA) / MeCN (0.1% TFA) = 100 / 0-50 / 50) and then freeze-dried to obtain N-{27-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-27-oxo-3,6,9,12,15,18,21,24-octaoxaheptacosan-1-yl}-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamide (trifluoroacetic acid) salt (25 mg) as a solid.

[0493] Example L4

[0494] DMP (112 mg) was added to a DCM (1 mL) solution of 1-(2-hydroxyethyl)-1H-pyrrole-2,5-dione (38 mg), and the mixture was stirred overnight at room temperature. DCM (2 mL) and DMP (112 mg) were added, and the mixture was stirred overnight at room temperature. Under ice cooling, 10% aqueous sodium sulfite solution, saturated aqueous sodium bicarbonate solution, and chloroform were added, and the mixture was extracted with chloroform using a phase separator, and concentrated under reduced pressure. 5-({2-methoxy-4-[(piperazin-1-yl)methyl]phenyl}methyl)-4-(pentylsulfanyl)-5H-pyrrolo[3,2-d]pyrimidin-2-amine (30 mg), THF (2 mL), acetic acid (0.01 mL), and sodium triacetoxyborohydride (70 mg) were added to the obtained residue, and the mixture was stirred overnight at room temperature. Under ice cooling, water and saturated sodium bicarbonate aqueous solution were added, and the mixture was extracted with a mixed solvent (chloroform / MeCN=5 / 1), and then the organic layer was dried over anhydrous MgSO4 and concentrated under reduced pressure. The obtained residue was purified by reverse phase silica gel column chromatography (ODS, water (0.1% TFA) / MeCN (0.1% TFA)=100 / 0-0 / 100) to obtain 1-(2-{4-[(4-{[2-amino-4-(pentylsulfanyl)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]piperazin-1-yl}ethyl)-1H-pyrrole-2,5-dione (trifluoroacetic acid) salt (13.2 mg) as a solid.

[0495] Production Example L5-1

[0496] Under ice cooling, a solution of triphosgene (593 mg) in DCM (4 mL) was added to a mixed solution of (2S)-2-(hydroxymethyl)pyrrolidine-1-carboxylic acid benzyl ester (0.94 g), DCM (6 mL) and DIPEA (2 mL), and the mixture was stirred at the same temperature for 3 hours. Then, tert-butyl (2-aminoethyl)carbamate (704 mg) was added at 0°C, and the reaction mixture was stirred at room temperature overnight. Saturated aqueous ammonium chloride solution (10 mL) was added, and the mixture was extracted twice with DCM (10 mL). The combined organic layer was dried over anhydrous MgSO4 and then concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate / hexane=5 / 95-80 / 20) to obtain benzyl (2S)-2-(10,10-dimethyl-3,8-dioxo-2,9-dioxa-4,7-diazaundec-1-yl)pyrrolidine-1-carboxylate (1.07 g) as an oily substance.

[0497] Production Example L5-2

[0498] Under a hydrogen atmosphere, a methanol (30 mL) solution of 10% palladium-supported carbon (50% water content, 50 mg) and (2S)-2-(10,10-dimethyl-3,8-dioxo-2,9-dioxa-4,7-diazaundecane-1-yl)pyrrolidine-1-carboxylic acid benzyl ester (1.05 g) was stirred at room temperature for 20 hours. The mixture was filtered through diatomaceous earth (registered trademark), the diatomaceous earth (registered trademark) was washed with methanol, and the filtrate was concentrated under reduced pressure and then dried to obtain ethane-1,2-diylbiscarbamic acid = tert-butyl = [(2S)-pyrrolidin-2-yl] methyl ester (710 mg) as an oily substance.

[0499] Production Example L5-3

[0500] To a solution of tert-butyl[(2S)-pyrrolidin-2-yl]methyl ethane-1,2-diylbiscarbamate (84 mg) and DIPEA (0.15 mL) in DMF (5 mL) was added 7-chloro-5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-N-[((2S)-pyrrolidin-2-yl)methyl]-1,2-diylbiscarbamate (84 mg) and DIPEA (0.15 mL) in DMF (5 mL). 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine hydrochloride (110 mg) was added and stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (amino silica gel, methanol / chloroform = 3 / 97) to obtain ethane-1,2-diylbiscarbamic acid = {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl = tert-butyl ester (87 mg) as a solid.

[0501] Example L5

[0502] 4M HCl / EtOAc (2 mL) was added to a solution of ethane-1,2-diylbiscarbamic acid = {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl = tert-butyl ester (84 mg) in ethyl acetate (2 mL), and the mixture was stirred at room temperature for 4 hours. After concentration under reduced pressure, the crude product was dissolved in THF (2 mL), and an aqueous sodium bicarbonate solution (1 mL) was added, followed by addition of methyl 2,5-dioxo-2,5-dihydro-1H-pyrrole-1-carboxylate (70 mg). The mixture was stirred at room temperature for 4 hours, and the reaction solution was directly purified by reverse phase silica gel column chromatography (ODS, MeCN / water = 0 / 100-100 / 0). The fractions containing the target product were recovered and purified again by silica gel column chromatography (methanol / chloroform = 0 / 100-10 / 90) to obtain [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamic acid {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl ester (6 mg) as a solid.

[0503] Production Example L6-1

[0504] Under ice-cold, (2S)-2-[(2-tert-butoxy-2-oxoethoxy)methyl]pyrrolidine-1-carboxylic acid tert-butyl ester (2.95g) was slowly added to a THF (50mL) solution of LAH (535mg), and then stirred at room temperature for 30 minutes. The reaction solution was ice-cold, water (0.5mL) was added, and then 1M aqueous sodium hydroxide solution (0.5mL) and anhydrous MgSO4 were added, and stirred for 30 minutes. After filtration, the filtrate was concentrated under reduced pressure to obtain (2S)-2-[(2-hydroxyethoxy)methyl]pyrrolidine-1-carboxylic acid tert-butyl ester (1.41g) as an oil.

[0505] Production Example L6-2

[0506] To a THF (10 mL) solution of (2S)-2-[(2-hydroxyethoxy)methyl]pyrrolidine-1-carboxylic acid tert-butyl ester (630 mg), triphenylphosphine (1.35 g) and phthalimide (755 mg) was added dropwise DEAD (2.2 mol / L, 2.4 mL) over 5 to 10 minutes. The reaction mixture was stirred for 16 hours and the precipitate was filtered. The filtrate was concentrated under reduced pressure and the crude product was purified by silica gel column chromatography (ethyl acetate / hexane=20 / 80) to obtain (2S)-2-{[2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)ethoxy]methyl}pyrrolidine-1-carboxylic acid tert-butyl ester (911 mg) as a solid.

[0507] Production Example L6-3

[0508] Under a nitrogen atmosphere, 4M HCl / EtOAc (4 mL) was added to a solution of tert-butyl (2S)-2-{[2-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)ethoxy]methyl}pyrrolidine-1-carboxylate (911 mg) in ethyl acetate (20 mL) under ice cooling. The reaction mixture was stirred at room temperature for 16 hours, and the generated solid was filtered to obtain 2-(2-{[(2S)-pyrrolidin-2-yl]methoxy}ethyl)-1H-isoindole-1,3(2H)-dione hydrochloride (751 mg) as a solid.

[0509] Production Example L6-4

[0510] To a solution of 2-(2-{[(2S)-pyrrolidin-2-yl]methoxy}ethyl)-1H-isoindole-1,3(2H)-dione hydrochloride (100 mg) in DMF (5 mL) were added DIPEA (0.22 mL), TBAI (6 mg) and 7-chloro-5-{[4-(chloromethyl)-2-methoxyphenyl]methyl}-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine hydrochloride (148 mg) was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the crude product was dissolved in ethanol (3 mL), and hydrazine monohydrate (35 mg) was added at 60 ° C. and stirred at the same temperature for 5 hours. The solid was separated by filtration using diatomaceous earth (registered trademark), washed with ethanol (50 mL), and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (amino silica gel, chloroform / methanol = 98:2) to obtain 5-{[4-({(2S)-2-[(2-aminoethoxy)methyl]pyrrolidin-1-yl}methyl-2-methoxyphenyl]methyl}-7-chloro-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (145 mg).

[0511] Example L6

[0512] To 5-{[4-({(2S)-2-[(2-aminoethoxy)methyl]pyrrolidin-1-yl}methyl-2-methoxyphenyl]methyl}-7-chloro-N 4 0.85M sodium bicarbonate aqueous solution (1.5mL) and 2,5-dioxo-2,5-dihydro-1H-pyrrole-1-carboxylic acid methyl ester (30mg) were added to a solution of -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine (140mg) in 1,4-dioxane (2mL), and stirred at the same temperature for 30 minutes. Water (10mL) was added to the reaction mixture, and it was extracted twice with chloroform / methanol (4:1, 10mL). The combined organic layer was dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol / chloroform=0 / 100-10 / 90) to obtain 1-[2-({(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methoxy)ethyl]-1H-pyrrole-2,5-dione (96 mg) as an oily substance.

[0513] Example LA: Preparation of Antibody Drug Conjugates

[0514] Common procedure A: Cleaning and concentration of antibody-drug complex aqueous solution

[0515] The antibody-drug complex solution was placed in an Amicon ultra-15 (10,000 MWCO, Millipore Co.) container, and D-PBS phosphate-buffered saline was added to make the solution volume 15 mL. The antibody-drug complex solution was then washed and concentrated by centrifugation (at 3,000 G for 15 to 90 minutes) using a centrifuge.

[0516] Common procedure B: Purification of antibody-drug conjugates

[0517] Use commercially available D-PBS (Cat. No. 045-29795, Fujifilm Wako) to balance 1 to 3 PD-10 columns. Load the antibody-drug complex reaction aqueous solution (about 300 μL to 2.5 mL) on each PD-10 column, elute with about 5 mL of D-PBS, and collect every 500 μL. Wash and concentrate the collected flow containing the antibody by repeating common operation A once or twice. Filter the recovered solution using a membrane filter to obtain an antibody-drug complex.

[0518] Common procedure C: Determination of drug concentration of antibody-drug complex

[0519] Drug concentration in antibody drug conjugates was measured using Thermo Scientific Pierce TM The calculation was performed by the BCA method using a BCA protein analysis kit, etc. A standard curve of absorbance at 562 nm was prepared using anti-human CLDN6 (hCLDN6) antibody not bound to the compound, and the drug concentration was calculated from the absorbance at 562 nm of the anti-hCLDN6-TLR7 / 8 dual agonist antibody-drug complex sample.

[0520] Common procedure D: Determination of DAR of anti-hCLDN6 antibody-drug complex containing TLR7 / 8 dual agonist

[0521] The DAR of the anti-hCLDN6-TLR7 / 8 dual agonist antibody drug complex is calculated by the deconvolution mass spectrum obtained by SEC-MS measurement. The deglycosylated antibody drug complex sample is injected into the LC / MS such as WatersXevo G2-XS connected to the Waters ACQUITY UPLC I-Class system. The measurement data is analyzed using Waters UNIFI. The spectrum intensity of each MS peak of the antibody-TLR7 / 8 dual agonist antibody drug complex is obtained based on the deconvolution mass spectrum obtained by the analysis, and the average DAR is calculated.

[0522] Common procedure E: Determination of the monomer purity of anti-hCLDN6 antibody-drug conjugates containing TLR7 / 8 dual agonists

[0523] The monomer purity of the anti-hCLDN6-TLR7 / 8 dual agonist antibody drug complex was calculated from the chromatogram obtained by SEC measurement. Using a YMC-SEC MAB column, 100mM potassium phosphate, 200mM sodium chloride buffer (pH7.0) / 2-propanol (85:15) was delivered at a flow rate of 0.165mL / min to elute the anti-hCLDN6-TLR7 / 8 dual agonist antibody drug complex, and the absorption of UV 280nm was detected. The integral value of each peak was obtained to calculate the monomer purity.

[0524] Example L1A1

[0525] The 13.57 mg / mL Ab1 antibody solution (147 μL) was collected into a 1.5 mL Eppendorf tube and the antibody concentration was adjusted to 2 mg / mL using PBS6.2 / EDTA (853 μL). Two identical antibody solutions were prepared in total, and 0.2 M sodium dihydrogen phosphate aqueous solution (51.5 μL) and 2 mM TCEP aqueous solution (31.9 μL) were added to each solution and incubated at 37°C for 1 hour. Next, 2 mM DMSO solution (63.8 μL) of 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazine-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione (trifluoroacetic acid) salt was added, and the mixture was allowed to stand at room temperature for 40 minutes. Next, 20 mM NAC aqueous solution (12.3 μL) was added, and the mixture was allowed to stand at room temperature for 20 minutes to stop the reaction between the drug and the linker. A total of two portions of the reaction solution were purified by common operation B to obtain a solution containing the antibody drug complex.

[0526] In addition, 13.57 mg / mL Ab1 antibody solution (147 μL) was collected in a 1.5 mL microcentrifuge tube, and the antibody concentration was adjusted to 2 mg / mL using PBS6.2 / EDTA (853 μL). A total of 3 identical antibody solutions were prepared, and 0.2 M sodium dihydrogen phosphate aqueous solution (51.5 μL) and 2 mM TCEP aqueous solution (31.9 μL) were added to each solution, and incubated at 37°C for 1 hour. Next, a 2 mM DMSO solution (63.8 μL) of 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazine-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione (trifluoroacetic acid) salt was added, and the mixture was allowed to stand at room temperature for 40 minutes. Next, a 20 mM NAC aqueous solution (12.3 μL) was added, and the mixture was allowed to stand at room temperature for 20 minutes to stop the reaction of the drug linker. A total of three portions of this reaction solution were purified by common operation B to obtain a solution containing an antibody drug complex.

[0527] After mixing the two solutions containing the antibody drug conjugate obtained above, washing and concentration were performed by repeating common operation A twice. The recovered solution was filtered with a membrane filter and diluted with D-PBS to a solution volume of 1.8 mL. Through common operations C and D, it was confirmed that the solution contained 3.88 mg / mL of the title antibody drug conjugate (DAR 3.44).

[0528] Example L1A2

[0529] The 14.72 mg / mL Ab2 antibody solution (135.9 μL) was collected into a 1.5 mL microcentrifuge tube and the antibody concentration was adjusted to 2 mg / mL using PBS6.2 / EDTA (864.1 μL). A total of 3 identical antibody solutions were prepared, and 0.2 M sodium dihydrogen phosphate aqueous solution (51.5 μL) and 2 mM TCEP aqueous solution (31.9 μL) were added to each solution and incubated at 37°C for 1 hour. Next, a 2 mM DMSO solution (63.8 μL) of 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione (trifluoroacetic acid) salt was added, and the mixture was allowed to stand at room temperature for 40 minutes. Next, a 20 mM NAC aqueous solution (12.3 μL) was added, and the mixture was allowed to stand at room temperature for 20 minutes to stop the reaction of the drug linker. A total of three portions of this reaction solution were purified by common operation B to obtain 1.25 mL of a solution containing the antibody drug conjugate. The solution was confirmed to contain 4.61 mg / mL of the title antibody drug conjugate (DAR4.02) by common operations C and D.

[0530] Example L5A1

[0531] The 13.57 mg / mL Ab1 antibody solution (147 μL) was collected into a 1.5 mL microcentrifuge tube and the antibody concentration was adjusted to 2 mg / mL using PBS6.2 / EDTA (853 μL). A total of 5 identical antibody solutions were prepared, and 0.2 M sodium dihydrogen phosphate aqueous solution (51.5 μL) and 2 mM TCEP aqueous solution (31.9 μL) were added to each solution and incubated at 37°C for 1 hour. Next, a 2 mM DMSO solution (63.8 μL) of {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamate was added, and the mixture was allowed to stand at room temperature for 40 minutes. Next, a 20 mM NAC aqueous solution (12.3 μL) was added, and the mixture was allowed to stand at room temperature for 20 minutes to stop the reaction of the drug linker. A total of 5 portions of this reaction solution were purified by common operation B to obtain 1.85 mL of a solution containing the antibody drug conjugate. The solution was confirmed to contain 4.55 mg / mL of the title antibody drug conjugate (DAR 2.95) by common operations C and D.

[0532] Example L7A1

[0533] A 13.57 mg / mL Ab1 antibody solution (147 μL) was collected in a 1.5 mL microcentrifuge tube and the antibody concentration was adjusted to 2 mg / mL using PBS6.2 / EDTA (853 μL). Two identical antibody solutions were prepared in total, and 0.2 M sodium dihydrogen phosphate aqueous solution (51.5 μL) and 2 mM TCEP aqueous solution (31.9 μL) were added to each solution, and incubated at 37°C for 1 hour. Next, a 2 mM DMSO solution of N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamide (trifluoroacetic acid) salt (63.8 μL) was added to each solution, and the mixture was allowed to stand at room temperature for 40 minutes. Next, a 20 mM NAC aqueous solution (12.3 μL) was added and the mixture was left to stand at room temperature for 20 minutes to terminate the reaction of the drug linker. A total of two portions of the reaction solution were purified by common procedure B to obtain a solution containing the antibody-drug conjugate.

[0534] Separately, a 13.57 mg / mL Ab1 antibody solution (147 μL) was collected in a 1.5 mL microcentrifuge tube, and the antibody concentration was adjusted to 2 mg / mL using PBS6.2 / EDTA (853 μL). A total of three identical antibody solutions were prepared, and 0.2 M sodium dihydrogen phosphate aqueous solution (51.5 μL) and 2 mM TCEP aqueous solution (31.9 μL) were added to each solution, and incubated at 37°C for 1 hour. Next, a 2 mM DMSO solution (63.8 μL) of N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamide (trifluoroacetic acid) salt was added, and the mixture was allowed to stand at room temperature for 40 minutes. Next, a 20 mM NAC aqueous solution (12.3 μL) was added and the mixture was left to stand at room temperature for 20 minutes to terminate the reaction of the drug linker. A total of three portions of this reaction solution were purified by common procedure B to obtain a solution containing the antibody-drug conjugate.

[0535] After mixing the two solutions containing the antibody-drug conjugate obtained above, washing and concentration were performed by repeating common operation A twice. The recovered solution was filtered with a membrane filter and diluted with D-PBS to a solution volume of 1.8 mL. Through common operations C and D, it was confirmed that the solution contained 4.52 mg / mL of the title antibody-drug conjugate (DAR 3.28).

[0536] The compounds or salts thereof, or antibody-drug conjugates or salts thereof described in the Preparation Examples and Examples shown in the following tables were produced by the methods described in the above Preparation Examples and Examples or in the same manner as the methods described therein.

[0537] [Table 3-1]

[0538]

[0539] [Table 3-2]

[0540]

[0541] [Table 3-3]

[0542]

[0543] [Table 3-4]

[0544]

[0545] [Table 4-1]

[0546]

[0547] [Table 4-2]

[0548]

[0549] [Table 4-3]

[0550]

[0551] [Table 5]

[0552]

[0553] [Table 6-1]

[0554]

[0555] [Table 6-2]

[0556]

[0557] [Table 6-3]

[0558]

[0559] Industrial Applicability

[0560] The antibody-drug conjugate of formula (I) or a salt thereof has an excellent TLR7 / 8 dual agonist effect, and also exerts TNF-α production effect, INF-γ production effect and in vivo anti-tumor effect, and is therefore expected to be used for the prevention and / or treatment of various cancers such as ovarian cancer, testicular cancer, cervical cancer, and lung cancer. In addition, the anti-CLDN6 antibody discovered in the present invention has binding activity to human CLDN6, and is expected to be used as a cancer prevention and / or therapeutic agent. Antibody-drug conjugates or salts thereof of formula (I) are expected to be used.

[0561] Sequence listing of independent text

[0562] Sequence number 2 is the amino acid sequence of the heavy chain of humanized anti-CLDN6 antibody Ab1, and the base sequence shown in sequence number 1 is a base sequence encoding the amino acid sequence shown in sequence number 2. Sequence number 4 is the amino acid sequence of the light chain of humanized anti-CLDN6 antibody Ab1, and the base sequence shown in sequence number 3 is a base sequence encoding the amino acid sequence shown in sequence number 4. Sequence number 6 is the amino acid sequence of the heavy chain of humanized anti-CLDN6 antibody Ab2, and the base sequence shown in sequence number 5 is a base sequence encoding the amino acid sequence shown in sequence number 6. Sequence number 8 is the amino acid sequence of the light chain of humanized anti-CLDN6 antibody Ab2, and the base sequence shown in sequence number 7 is a base sequence encoding the amino acid sequence shown in sequence number 8.

Claims

1. An antibody-drug complex of formula (I) or a salt thereof: [Chemical formula 1] In the formula, Ab is an antibody or its antigen-binding fragment, X is S or NH, R 1 is a lower alkyl group or -CH2-isoxazolediyl-CH3, Here, in R 1 In the case of -CH2-isoxazolediyl-CH3, X is S, R 2 is halogen or H, R 3 is CH3 or H, R 4 is a group represented by formula (a), formula (b) or formula (c), [Chemical formula 2] Ring A is a 4- to 6-membered cyclic amine consisting of multiple Cs and 1 N. Ring B is a 4- to 6-membered cyclic amine composed of multiple Cs, 1 N and 1 G, G is N or CH, R A is -CH2O-, C(CH3)2O-, -O- or -CH2NH-, Here, in R A When it is -O- or -CH2NH-, X is S; R 2 is halogen; or R 3 For CH3, R B is a halogenated alkyl, CH2OH, C(CH3)2OH, OH, CH2NH2 or H, Here, in R B When OH, CH2NH2 or H is present, G is CH; X is S; R 2 is halogen; or R 3 For CH3, R C C 3-6 Cycloalkyl, or -CH2-C 3-6 Cycloalkyl, L is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, -(CH2CH2O) m -C(=O)-lower alkylene-, or -C(=O)-cyclohexanediyl-lower alkylene-, m is an integer from 1 to 10, n is 1 to 16.

2. The antibody-drug conjugate or a salt thereof according to claim 1, wherein Ring A is a 5-membered cyclic amine composed of 4 Cs and 1 N, and Ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs, and 1 G, wherein G is N.

3. The antibody-drug conjugate or a salt thereof according to claim 2, wherein: R A is -CH2O- or -C(CH3)2O-, R B It is halogenated alkyl, CH2OH or C(CH3)2OH.

4. The antibody-drug conjugate or a salt thereof according to claim 3, wherein X is NH, R 1 is a lower alkyl group, R 3 For H.

5. The antibody-drug conjugate or a salt thereof according to claim 4, wherein L is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, or -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, m is an integer of 4-8.

6. The antibody-drug conjugate or a salt thereof according to claim 5, wherein L is -lower alkylene-, -C(=O)lower alkylene-, or -C(=O)NHlower alkylene-.

7. The antibody-drug conjugate or a salt thereof according to claim 1, wherein X is NH, R 1 is a lower alkyl group, R 2 is halogen, R 3 For H, R 4 Formula (a), Ring A is a 5-membered cyclic amine consisting of 4 Cs and 1 N, R A is -CH2O-, and L is -C(=O)NHCH2CH2-.

8. The antibody-drug conjugate or a salt thereof according to claim 1, wherein X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 Formula (b), Ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, G is N, R B is a halogenated alkyl group, and L is -C(=O)CH2CH2-.

9. The antibody-drug conjugate or a salt thereof according to claim 1, wherein X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 Formula (c), R C C 3-6 Cycloalkyl, L is -C(=O)CH2CH2-.

10. The antibody-drug conjugate or a salt thereof according to claim 1, wherein The antibody drug conjugate of formula (I) is selected from the group consisting of the following antibody drug conjugates: Antibody-drug complex formed by binding Ab to 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazine-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione, An antibody-drug complex formed by binding Ab to [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamic acid {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl ester, An antibody-drug complex in which Ab is conjugated with N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamide, and The antibody-drug complex is formed by binding Ab to 1-{3-[(3R)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione.

11. The antibody-drug conjugate of formula (I) or a salt thereof according to any one of claims 1 to 10, wherein: Ab is an anti-TAA antibody or an antigen-binding fragment thereof selected from anti-A33 antibody, anti-B7-H3 antibody, anti-CanAg antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD30 antibody, anti-CD33 antibody, anti-CD56 antibody, anti-CD70 antibody, anti-CEA body, anti-CLDN6 antibody, anti-EphA2 antibody, anti-G250 antibody, anti-MUC1 antibody, anti-GPNMB antibody, anti-Integrin antibody, anti-PSMA antibody, anti-Tenascin-C antibody, anti-TROP-2 antibody, and anti-TSPAN8 antibody.

12. The antibody-drug conjugate or a salt thereof according to claim 11, wherein Ab is an anti-CLDN6 antibody or an antigen-binding fragment thereof.

13. The antibody-drug conjugate or a salt thereof according to claim 12, wherein Ab is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of Ab-A and Ab-B, Ab-A is an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acid numbers 31 to 35 of SEQ ID NO: 2, a CDR2 consisting of an amino acid sequence of amino acid numbers 50 to 65 of SEQ ID NO: 2, and a CDR3 consisting of an amino acid sequence of amino acid numbers 95 to 102 of SEQ ID NO: 2, and the light chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acid numbers 24 to 34 of SEQ ID NO: 4, a CDR2 consisting of an amino acid sequence of amino acid numbers 50 to 56 of SEQ ID NO: 4, and a CDR3 consisting of an amino acid sequence of amino acid numbers 89 to 97 of SEQ ID NO: 4, Ab-B is an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acid numbers 31 to 35 of SEQ ID NO: 6, a CDR2 consisting of an amino acid sequence of amino acid numbers 50 to 65 of SEQ ID NO: 6, and a CDR3 consisting of an amino acid sequence of amino acid numbers 95 to 102 of SEQ ID NO: 6, and the light chain variable region comprises: a CDR1 consisting of an amino acid sequence of amino acid numbers 24 to 34 of SEQ ID NO: 8, a CDR2 consisting of an amino acid sequence of amino acid numbers 50 to 56 of SEQ ID NO: 8, and a CDR3 consisting of an amino acid sequence of amino acid numbers 89 to 97 of SEQ ID NO:

8.

14. The antibody-drug conjugate or a salt thereof according to claim 12, wherein Ab is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of Ab-C and Ab-D, Ab-C is an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of the amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 2 and a light chain variable region consisting of the amino acid sequence of amino acids 1 to 108 of SEQ ID NO: 4, Ab-D is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of the amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 6, and a light chain variable region consisting of the amino acid sequence of amino acids 1 to 108 of SEQ ID NO:

8.

15. The antibody-drug conjugate or a salt thereof according to claim 12, wherein Ab is an anti-CLDN6 antibody or an antigen-binding fragment thereof selected from the group consisting of Ab1 and Ab2, Ab1 is an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO: 2 and a light chain consisting of the amino acid sequence of SEQ ID NO: 4, Ab2 is an anti-CLDN6 antibody comprising a heavy chain consisting of the amino acid sequence of SEQ ID NO:6 and a light chain consisting of the amino acid sequence of SEQ ID NO:

8.

16. The antibody-drug conjugate or a salt thereof according to claim 1, wherein Ab is the anti-CLDN6 antibody Ab1, X is NH, R 1 is a lower alkyl group, R 2 is halogen, R 3 For H, R 4 Formula (a), Ring A is a 5-membered cyclic amine consisting of 4 Cs and 1 N, R A is -CH2O-, and L is -C(=O)NHCH2CH2-.

17. The antibody-drug conjugate or a salt thereof according to claim 1, wherein Ab is the anti-CLDN6 antibody Ab1, X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 Formula (b), Ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, G is N, R B is a halogenated alkyl group, and L is -C(=O)CH2CH2-.

18. The antibody-drug conjugate or a salt thereof according to claim 1, wherein Ab is the anti-CLDN6 antibody Ab1, X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 4 Formula (c), R C C 3-6 Cycloalkyl, L is -C(=O)CH2CH2-.

19. The antibody-drug conjugate or a salt thereof according to claim 1, wherein The antibody drug conjugate is selected from the group consisting of: An antibody-drug complex formed by binding Ab1 to 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione, An antibody-drug complex formed by binding Ab2 to 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione, An antibody-drug complex formed by binding Ab1 to [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamic acid {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl ester, An antibody-drug complex in which Ab1 is bound to N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamide, and The antibody-drug complex is formed by the binding of Ab1 to 1-{3-[(3R)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione.

20. The antibody-drug conjugate or a salt thereof according to claim 19, wherein The antibody drug conjugate is selected from the group consisting of: [Chemical formula 3] n is 1 to 16.

21. A pharmaceutical composition comprising the antibody-drug conjugate or a salt thereof according to claim 1 and a pharmaceutically acceptable excipient.

22. The pharmaceutical composition according to claim 21, which is a pharmaceutical composition for preventing or treating cancer.

23. Use of the antibody-drug conjugate or a salt thereof according to claim 1 for producing a pharmaceutical composition for preventing or treating cancer.

24. Use of the antibody-drug conjugate or a salt thereof according to claim 1 for preventing or treating cancer. The antibody-drug conjugate or a salt thereof according to claim 1, which is used for the prevention or treatment of cancer.

26. A method for preventing or treating cancer, comprising administering an effective amount of the antibody-drug conjugate or a salt thereof according to claim 1 to a subject.

27. A compound of formula (II) or a salt thereof: [Chemical formula 4] In the formula, X is S or NH, R 1 is a lower alkyl group or -CH2-isoxazolediyl-CH3, Here, in R 1 In the case of -CH2-isoxazolediyl-CH3, X is S, R 2 is halogen or H, R 3 is CH3 or H, R 40 is a group represented by formula (a1), formula (b1) or formula (c1), [Chemical formula 5] Ring A is a 4- to 6-membered cyclic amine composed of multiple C atoms and one N atom. Ring B is a 4- to 6-membered cyclic amine composed of multiple Cs, 1 N and 1 G, G is N or CH, R A is -CH2O-, -C(CH3)2O-, -O-, or -CH2NH-, Here, in R A When it is -O- or -CH2NH-, X is S; R 2 is halogen; or R 3 For CH3, R B is a haloalkyl, CH2OH, C(CH3)2OH, OH, CH2NH2, or H, Here, in R B When OH, CH2NH2 or H is present, G is CH; X is S; R 2 is halogen; or R 3 For CH3, R C C 3-6 Cycloalkyl or -CH2-C 3-6 Cycloalkyl, M is H, or a group represented by formula (d): [Chemical formula 6] L D -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, -(CH2CH2O) m -C(=O)-lower alkylene-, or -C(=O)-cyclohexanediyl-lower alkylene-, m is an integer of 1-10.

28. The compound or salt thereof according to claim 27, wherein Ring A is a 5-membered cyclic amine consisting of 4 Cs and 1 N. Ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs, and 1 G.

29. The compound or salt thereof according to claim 28, wherein R A is -CH2O- or -C(CH3)2O-, R B It is halogenated alkyl, CH2OH or C(CH3)2OH.

30. The compound or salt thereof according to claim 29, wherein L D is -lower alkylene-, -C(=O)lower alkylene-, -C(=O)NHlower alkylene-, or -C(=O)lower alkylene-(OCH2CH2) m -NH-C(=O)lower alkylene-, m is an integer of 4-8.

31. The compound or salt thereof according to claim 30, wherein X is NH, R 1 is a lower alkyl group, R 2 is halogen or H, R 3 For H.

32. The compound or salt thereof according to claim 27, wherein X is NH, R 1 is a lower alkyl group, R 2 is Cl or H, R 3 For H, R 40 The compound is of formula (a1), formula (b1) or formula (c1), wherein ring A is a 5-membered cyclic amine composed of 4 Cs and 1 N, and ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, wherein G is N, and R A is -CH2O-, R B is a halogenated alkyl group, R C C 3-6 cycloalkyl, M is a group represented by formula (d), L D It is -C(=O)CH2CH2- or -C(=O)NHCH2CH2-.

33. The compound or salt thereof according to claim 27, wherein X is NH, R 1 is a lower alkyl group, R 2 For H, R 3 For H, R 40 Formula (b1), Ring B is a 6-membered cyclic amine composed of 2 Cs, 1 N, 2 Cs and 1 G, G is N, R B is a halogenated alkyl group, L D It is -C(=O)CH2CH2-.

34. The compound according to claim 27 or a salt thereof, which is selected from the group consisting of: 5-[(4-{[(2S)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof, 5-[(4-{[(3R)-3-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof, 5-({4-[(cyclopropylamino)methyl]-2-methoxyphenyl}methyl)-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof, {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methanol or a salt thereof, 5-({2-methoxy-4-[(piperidin-4-yl)methyl]phenyl}methyl)-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof, 5-[(4-{[(2R)-2-(fluoromethyl)piperazin-1-yl]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof, 5-[(4-{[(cyclobutylmethyl)amino]methyl}-2-methoxyphenyl)methyl]-N 4 -pentyl-5H-pyrrolo[3,2-d]pyrimidine-2,4-diamine or a salt thereof, 1-{3-[(3S)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione or a salt thereof, {(2S)-1-[(4-{[2-amino-7-chloro-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]pyrrolidin-2-yl}methyl [2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl]carbamate or a salt thereof, N-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-N-cyclopropyl-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamide or a salt thereof, and 1-{3-[(3R)-4-[(4-{[2-amino-4-(pentylamino)-5H-pyrrolo[3,2-d]pyrimidin-5-yl]methyl}-3-methoxyphenyl)methyl]-3-(fluoromethyl)piperazin-1-yl]-3-oxopropyl}-1H-pyrrole-2,5-dione or a salt thereof.

35. An anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 2, and a light chain variable region consisting of amino acids 1 to 108 of SEQ ID NO: 4; or, an anti-CLDN6 antibody or an antigen-binding fragment thereof comprising a heavy chain variable region consisting of an amino acid sequence of amino acids 1 to 117 of SEQ ID NO: 6, and a light chain variable region consisting of amino acids 1 to 108 of SEQ ID NO: 8.

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