PTK7-binding materials, their complexes, and methods for using them

Abstract

The present invention provides PTK7 antibodies, their antigen-binding moieties, other binding substances, and their PTK7 complexes, as well as methods for the treatment of cancer and autoimmune diseases, and the use of such antibodies and complexes.

Description

TECHNICAL FIELD

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of International Patent Application No. PCT / CN2023 / 121540 filed on September 26, 2023, and International Patent Application No. PCT / CN2024 / 104820. Each of the said applications is incorporated herein by reference in its entirety.

[0002] SEQUENCE LISTING This application contains a Sequence Listing which has been submitted electronically in XML file format and is incorporated herein by reference in its entirety. The XML copy created on September 26, 2024 is named GMI-408_SL.xml and has a size of 52.7KB.

[0003] TECHNICAL FIELD The present disclosure relates generally to antibodies and antibody-drug conjugates, as well as methods of using the antibodies and antibody-drug conjugates, and in particular to such antibodies, antibody-drug conjugates, and methods related to PTK7-expressing diseases and disorders.

BACKGROUND ART

[0004] BACKGROUND There has been considerable interest in using monoclonal antibodies (mAbs) in the form of antibody-drug conjugates (ADCs) for targeted delivery of cytotoxic substances to disease-associated cells, such as cancer cells and other cells. Designing antibody-drug conjugates, which involve conjugating cytotoxic substances, immunomodulators, or other drugs (collectively, "drugs") to antibodies, typically via linkers, requires consideration of various factors. These factors include the identification and positioning of chemical groups for drug binding, the mechanism of drug release, any structural elements that lead to drug release, and, if any, the structural changes of the released free drug. If the drug is released into the extracellular environment, the released form of the drug must be able to reach its target. If the drug is released after internal translocation of the antibody-drug conjugate, the structural elements and mechanism of drug release must be in harmony with the intracellular transport of the conjugate.

[0005] Another important factor in antibody-drug conjugate design is the amount of drug that can be delivered per targeting substance (i.e., the number of drugs bound to each targeting substance (e.g., antibody), called the drug content or drug load). Historically, it was believed that a higher drug content was superior to a lower content (e.g., 8 content vs. 4 content). The rationale was that a higher conjugate content would deliver more drug (e.g., cytotoxic substances) to target cells. This rationale was supported by observations that conjugates with higher drug loads showed higher activity against cell lines in vitro. However, subsequent studies have shown that this hypothesis is not confirmed in animal models. Conjugates with drug content of 4 or 8 of a particular auristatin were observed to have similar activity in mouse models. See, for example, Hamblett et al., Clinical Cancer Res. 10:7063-70 (2004) (Non-Patent Literature 1). Hamblett et al. further reported that in animal models, higher ADC content leads to more rapid elimination from the circulating blood. This more rapid elimination suggests that high-content formulations present PK challenges compared to low-content formulations. See Hamblett et al. Furthermore, higher-content conjugates resulted in lower maximum tolerated dose (MTD) in mice, and consequently, narrower reported therapeutic indices. Ibid. In contrast, ADCs with a drug loading of 2 at the manipulated site within the monoclonal antibody have been reported to have the same or better PK and therapeutic indices compared to certain ADCs with a loading of 4. See, for example, Junutula et al., Clinical Cancer Res. 16:4769 (2010) (Non-Patent Literature 2). Therefore, the recent trend is to develop ADCs with lower drug loadings.

[0006] An attractive target for ADC-based cancer therapies is protein tyrosine kinase 7 (PTK7). Also known as colon cancer kinase 4 (CCK4), PTK7 is a member of the receptor tyrosine kinase protein family that transmits extracellular signals across the cell membrane. Although this protein lacks detectable catalytic tyrosine kinase activity, it is involved in the Wnt signaling pathway and plays a role in multiple cellular processes, including polarity and adhesion.

[0007] PTK7 is known to be highly expressed in many types of cancer, but its expression is limited in normal human tissue. This makes PTK7 an attractive target for cancer treatment. However, the pace of developing effective PTK7 antibodies and related conjugates has been slow, and clinical trials using PTK7 antibodies and PTK7 ADCs have so far yielded only limited results.

[0008] Therefore, there is a need for PTK7 antibodies in general, and especially for PTK7 ADCs, that allow for higher drug loadings while maintaining other properties of the conjugate at lower loadings, such as preferred PK properties. Aspects of the present invention address these and related needs. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Hamblett et al., Clinical Cancer Res. 10:7063-70 (2004) [Non-Patent Document 2] Junutula et al., Clinical Cancer Res. 16:4769 (2010) [Overview of the project]

[0010] overview This specification provides PTK7 conjugates, antibody-drug conjugates (ADCs), and methods of using the conjugates and ADCs to treat diseases such as cancer and autoimmune diseases.

[0011] In some cases, It includes a heavy chain variable (VH) region and a light chain variable (VL) region, The VH region includes complementarity determination regions HCDR1, HCDR2, and HCDR3 located within the heavy chain variable region framework region. The VL region includes LCDR1, LCDR2, and LCDR3 located in the light chain variable region framework region. VH and VL CD-Rs These are SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, FAS, and SEQ ID NO:7, respectively. These are SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, RTS, and SEQ ID NO:14, respectively. These are SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, KVS, and SEQ ID NO:22, respectively. Having an amino acid sequence selected from the set of amino acid sequences shown in the group consisting of, A binding agent is provided.

[0012] In some embodiments, pharmaceutical compositions comprising the binding substance of this disclosure and a pharmaceutically acceptable carrier are provided herein.

[0013] In some embodiments, nucleic acids encoding the binding material of the present disclosure are provided herein.

[0014] In some embodiments, vectors comprising nucleic acids described in this disclosure are provided herein.

[0015] In some embodiments, provided herein is a vector comprising the nucleic acid of the present disclosure.

[0016] In some embodiments, provided herein is a cell line comprising the binding agent, vector, or nucleic acid of the present disclosure.

[0017] In some embodiments, provided herein is a conjugate comprising a binding agent, at least one linker bound to the binding agent, and at least one drug unit, wherein each drug unit is bound to the linker, and the linker may comprise at least one polar group.

[0018] In some embodiments, for the conjugate of the present disclosure, the linker is derived from a linker compound, or a stereoisomer or salt thereof, the linker compound comprises a linker unit, a stretcher group connected to the linker unit, an optional amino acid unit, and at least one polar group, where the stretcher group has a binding site to the binding agent and a binding site to the amino acid unit (if present) or to the linker subunit, the amino acid unit (if present) has a binding site to the stretcher unit and a binding site to the linker unit, the linker unit has a binding site to the amino acid unit (if present) or to the stretcher unit and to at least one drug unit.

[0019] In some embodiments, for the conjugate of the present disclosure, the linker unit is non-cleavable.

[0020] In some embodiments, for the conjugate of the present disclosure, the linker unit is cleavable by comprising a cleavable group.

[0021] In some embodiments, for the conjugate of the present disclosure, the polar group is bound to the amino acid unit.

[0022] In some embodiments, the polar group is bonded to the stretcher group in the complex of the present disclosure.

[0023] In some embodiments, the linker compound in the composite of the present disclosure is (a) Having 1 to 4 binding sites for drug units, and having the following structure (i) or (ii): A linker unit having one of TIFF2026534278000001.tif37128, (b) Each containing a polymer unit, at least one polar group, (c) A stretcher group having a binding site for a binding substance and Includes, During the ceremony, α- is a binding site to a group that can be cleaved by enzymes. β- is a binding site to at least one polar group, δ- is a binding site to H, at least one of the drug units, or a binding site to a linking group attached to at least one of the drug units. The polymer units include polyamides, polyethers, or combinations thereof, where the polyethers include hydroxyl groups, polyhydroxyl groups, sugar groups, carboxyl groups, or combinations thereof. Each R a These are independently H or C1-C6 alkyl groups. Each R b They are independent of Haro, C 1~6 A binding site to at least one alkyl group, a drug unit, or a binding site to at least one polar group, x is 0, 1, 2, 3, or 4. y is 0, 1, 2, or 3. R c The bonds are -C(O)-, -S(O)-, -SO2-, C 1~6 Alkilen, C 1~6 Alkinylene, triazolyl, or a combination thereof. Y is a bond, -O-, -S-, -N(R aThese are -C(O)-, -S(O)-, -SO2-C1~C6 alkylenes, C1~C6 alkenylenes, C1~C6 alkynylenes, triazolyls, or combinations thereof.

[0024] In some embodiments, pharmaceutical compositions comprising the complex of the present disclosure and a pharmaceutically acceptable carrier are provided herein.

[0025] In some embodiments, methods for treating PTK7+ cancer are provided herein, comprising the step of administering a therapeutically effective amount of the binding substance, pharmaceutical composition, complex, or pharmaceutical composition of the present disclosure to a subject in need.

[0026] In some embodiments, the use of the complex or pharmaceutical composition of this disclosure for the treatment of PTK7+ cancer in a subject is provided herein.

[0027] Further features are partially described below and will be partially apparent to those skilled in the art upon examination of the following and accompanying drawings, or may be learned through the manufacture or operation of the embodiments. The features of this disclosure can be realized and achieved by the implementation or use of various aspects of the methods, apparatus, and combinations described in the detailed embodiments discussed below. [Brief explanation of the drawing]

[0028] This disclosure will be further described based on exemplary embodiments. These exemplary embodiments are described in detail with reference to the drawings. Note that the drawings are not to scale. These embodiments are non-limiting and exemplary, and similar reference numerals throughout multiple drawings represent similar structures. [Figure 1] This graph shows the results of ELISA binding assays for humanized antibodies 2C5, 2C8, 13C4, and cofetuzumab against human PTK7. [Figure 2] This graph shows the results of ELISA binding assays for humanized antibodies 2C5, 2C8, 13C4, and cofetuzumab against cynomolgus monkey PTK7. [Figure 3] This graph shows the results of ELISA binding assays for humanized antibodies 2C5, 2C8, and 13C4 against mouse PTK7, as well as cofetuzumab. [Figure 4] This graph shows the results of ELISA binding assays of humanized antibodies 2C5, 2C8, 13C4, and cofetuzumab against rat PTK7. [Figure 5] This graph shows the antibody binding activity to the PA-1 cell line as measured by FACS. [Figure 6] This graph shows the binding activity of antibodies to the cell line MDA-MB-468 as measured by FACS. [Figure 7] This graph shows the binding activity of antibodies to the cell line MDA-MB-231 as measured by FACS. [Figure 8] This graph shows the binding activity of antibodies to the cell line MDA-MB-453 as measured by FACS. [Figure 9] This graph shows the internal transfer rates of the antibody 2C5 and cofetuzumab in the PA-1 cell line. [Figure 10] This graph shows the internal transfer rates of antibody 2C5 and cofetuzumab in the cell line MDA-MB-468. [Figure 11] This graph shows the internal transfer rates of antibody 2C5 and cofetuzumab in the cell line MDA-MB-453. [Figure 12] This graph shows the internal transfer rates of antibody 2C5 and cofetuzumab in the cell line MDA-MB-231. [Figure 13] This graph shows the internal transfer rates of the antibody 2C5 and cofetuzumab in the OVCAR-3 cell line. [Figure 14] This graph shows the internal transfer rates of the antibody 2C8 and cofetuzumab in the PA-1 cell line. [Figure 15] This graph shows the internal transfer rates of antibody 2C8 and cofetuzumab in the cell line MDA-MB-468. [Figure 16]This graph shows the internal transfer rates of antibody 2C8 and cofetuzumab in the cell line MDA-MB-453. [Figure 17] This graph shows the internal transfer rates of antibody 2C8 and cofetuzumab in the cell line MDA-MB-231. [Figure 18] This graph shows the internal transfer rates of the antibody 2C8 and cofetuzumab in the OVCAR-3 cell line. [Figure 19] This graph shows the internal transfer rates of antibody 13C4 and cofetuzumab in the PA-1 cell line. [Figure 20] This graph shows the internal transfer rates of antibody 13C4 and cofetuzumab in the cell line MDA-MB-468. [Figure 21] This graph shows the internal transfer rates of antibody 13C4 and cofetuzumab in the cell line MDA-MB-453. [Figure 22] This graph shows the internal transfer rates of antibody 13C4 and cofetuzumab in the cell line MDA-MB-231. [Figure 23] This graph shows the internal transfer rates of the antibody 13C4 and cofetuzumab in the OVCAR-3 cell line. [Figure 24] This graph shows the internal migration rate of 2C8 in various tumor cell lines. [Figure 25] This graph shows the internal migration rate of PRO1107 in various tumor cell lines. [Figure 26] This graph shows the cytotoxicity of ADCs containing the linker drug LD110 against the PA-1 cell line. [Figure 27] This graph shows the cytotoxicity of ADCs containing the linker drug LD110 against the cell line MDA-MB-468. [Figure 28] This graph shows the cytotoxicity of ADCs containing the linker drug LD110 against the cell line MDA-MB-453. [Figure 29] This graph shows the cytotoxicity of ADCs containing the linker drug LD110 against the OVCAR-3 cell line. [Figure 30]This graph shows the cytotoxicity of ADCs containing the linker drug LD038 against the tumor cell line OVCAR-3. [Figure 31] This graph shows in vitro cytotoxicity of the cell line MDA-MB-468 by PRO1107, MMAE, and b12-LD343(8). [Figure 32] This graph shows in vitro cytotoxicity of the cell line OVCAR-3 by PRO1107, MMAE, and b12-LD343(8). [Figure 33] This graph shows in vitro cytotoxicity of the cell line PA-1 by PRO1107, MMAE, and b12-LD343(8). [Figure 34] This graph shows in vitro cytotoxicity of the cell line Detroit 562 by PRO1107, MMAE, and b12-LD343(8). [Figure 35] This graph shows in vitro cytotoxicity of the AGS cell line by PRO1107, MMAE, and b12-LD343(8). [Figure 36] This graph shows in vitro cytotoxicity of the cell line SNG-M by PRO1107, MMAE, and b12-LD343(8). [Figure 37] This graph shows in vitro cytotoxicity of the cell line NCI-H292 by PRO1107, MMAE, and b12-LD343(8). [Figure 38] This graph shows in vitro cytotoxicity of the cell line NCI-H520 by PRO1107, MMAE, and b12-LD343(8). [Figure 39] This graph shows in vitro cytotoxicity of cell line SW780 by PRO1107, MMAE, and b12-LD343(8). [Figure 40] This graph shows the pharmacokinetics (PK) of antibodies 2C5, 2C8, 13C4, and cofetuzumab in rats. [Figure 41] This graph shows the pharmacokinetics (PK) of the antibodies 2C8 and ADC PRO1107 in rats. [Figure 42]This graph shows the efficacy of the 2C8 complex and cofetuzumab complex in an MDA-MB-468 xenograft model. [Figure 43] This graph shows the efficacy of the 2C8 complex and cofetuzumab complex in a PA-1 xenograft model. [Figure 44] This graph shows the efficacy of other 2C8 complexes and cofetuzumab complexes in PA-1 xenograft models. [Figure 45] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in a PA-1 xenograft model. [Figure 46] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in an OVCAR-3 xenograft model. [Figure 47] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in an MDA-MB-468 xenograft model. [Figure 48] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in the Detroit 562 xenograft model. [Figure 49] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in the SW780 xenograft model. [Figure 50] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in an RT4 xenograft model. [Figure 51] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in an AGS xenograft model. [Figure 52] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in a KYSE-150 xenograft model. [Figure 53] This graph shows the in vivo efficacy results of PRO1107, cofetuzumab peridotin, and 2C8-vedotin (4) in the NCI-H292 xenograft model. [Figure 54] Figure 54A is a graph showing the free MMAE of PRO1107, 2C8-vedotin(4), and cofetuzumab peridotin in tumors. Figure 54B is a graph showing the complexed MMAE of PRO1107, 2C8-vedotin(4), and cofetuzumab peridotin in tumors. [Figure 55] This shows the changes in body weight of rats after treatment with PRO1107, 2C8-vedotin(4), and cofetuzumab peridotin. [Figure 56] This graph shows the antitumor activity of single doses of mAb-LD343(8) and mAb-vedotin(4) in a PA-1 xenograft model. [Figure 57] This graph shows the antitumor activity of single doses of mAb-LD343(8) and mAb-vedotin(4) in a SW780 xenograft model. [Figure 58] This graph shows the antitumor activity of single doses of mAb-LD343(8) and mAb-vedotin(4) in an OVCAR-3 xenograft model. [Figure 59] This graph shows the antitumor activity of single doses of mAb-LD343(8) and mAb-vedotin(4) in the Detroit 562 xenograft model. [Figure 60] PRO1107 exhibits hydrophilicity. [Figure 61] This study demonstrates bystander effects of PRO1107 and vedotin-based ADCs in the cell lines PA-1 and THP-1-Luc. [Figure 62A] The graph shows the tumor weight changes in nude mice after treatment with PRO1107, 2C8-vedotin(4), and cofetuzumab peridotin. [Figure 62B] This shows the change in DAR value of PRO1107 during human plasma incubation. [Figure 62C]This shows the released payload (MMAE) at various time points in plasma from different species. [Figure 63] This study demonstrates the efficacy of PRO1107 in various cell-derived xenotransplant (CDX) models, including esophageal squamous cell carcinoma (ESCC), ovarian cancer (OVCA), breast cancer (BC), bladder cancer (BLC), gastric cancer (GC), lung cancer (LC), and head and neck squamous cell carcinoma (HNSCC). [Figure 64] This study demonstrates the efficacy of PRO1107 in various patient-derived xenotransplant (CDX) models, including bladder cancer (BLC), esophageal cancer (EsC), and uterine cancer (also known as endometrial cancer). [Modes for carrying out the invention]

[0029] Detailed explanation The following explanations are provided to enable any person skilled in the art to create and use this disclosure, and are provided in the context of a particular use and its requirements. Various modifications to the disclosed embodiments will be readily apparent to a person skilled in the art, and the general principles set forth herein may be applied to other embodiments and uses without departing from the spirit and scope of this disclosure. Thus, this disclosure is not limited to the embodiments shown and encompasses the broadest possible scope that is consistent with the claims.

[0030] The terms used herein are intended to describe specific exemplary embodiments only and are not intended to be limiting. As used herein, the singular forms “a,” “an,” and “the” may also be intended to include the plural form unless the context clearly indicates otherwise. It will be further understood that, as used herein, the terms “comprises,” “comprising,” “includes,” and / or “including” identify the presence of a described feature, integer, process, operation, element, and / or component, but do not exclude the presence or addition of one or more other features, integers, processes, operations, elements, components, and / or groups thereof. When an embodiment is described using the expression “comprises,” an embodiment that “essentially consists of” what is described is also provided. Furthermore, when an embodiment is described using the expression “comprises,” an embodiment that “consists of” what is described is also provided.

[0031] These and other features, the characteristics of this disclosure, the methods and functions of the related components, and the economics of combining and manufacturing the parts may become more apparent by considering the following description with reference to the accompanying drawings, all of which constitute part of this specification. However, it should be clearly understood that the drawings are for illustrative and illustrative purposes only and are not intended to limit the scope of this disclosure. The drawings are understood not to scale.

[0032] definition For convenience, specific terms used herein, in the examples and claims are defined herein. Unless otherwise specified or implied by the context, the following terms and expressions have the meanings provided below. The definitions are provided to aid in describing specific aspects and are not intended to limit the claimed invention, as the scope of the invention is limited only by the claims. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as they would normally be interpreted by those skilled in the art in which the invention pertains.

[0033] As used herein, and unless otherwise specified, the terms “a” and “an” mean “one,” “at least one,” or “one or more.” Unless the context requires otherwise, singular terms used herein shall include plural forms, and plural terms shall include singular forms.

[0034] Unless otherwise clearly required by context, throughout the specification and claims, phrases such as “includes” and “contains” shall be interpreted in a comprehensive sense, not exclusive or exhaustive, i.e., “non-restrictively include ~”.

[0035] The terms “decreased,” “decrease,” “decrease,” “decrease,” and “inhibit” are used herein to generally mean a statistically significant decrease relative to a reference value.

[0036] The terms “increased,” “enhance,” “improve,” or “activate” are used herein to generally mean an increase of a statistically significant amount relative to a baseline.

[0037] As used herein, the terms “protein” and “polypeptide” are used interchangeably to refer to a series of amino acid residues linked to one another by peptide bonds between the α-amino and carboxyl groups of adjacent residues. The terms “protein” and “polypeptide” also refer to polymers of amino acids, including modified amino acids (e.g., phosphorylated, glycated, glycosylated, etc.) and amino acid analogs, regardless of their size or function. While the terms “protein” and “polypeptide” are used for relatively large polypeptides, the term “peptide” is used for small polypeptides, but the usage of these terms overlaps in the art. The terms “protein” and “polypeptide” are used interchangeably herein when referring to encoded gene products and their fragments. Thus, exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologues, paralogs, fragments, and other equivalents, variants, fragments, and analogs of those described above.

[0038] PTK7 (protein tyrosine kinase-like 7) is a member of the receptor protein tyrosine kinase family and is also known as colon cancer kinase 4 (CCK4). In humans, this protein is encoded by the PTK7 gene. PTK7 is overexpressed on the surface of many types of cancer, including solid tumors indefinitely. Such solid tumors include breast cancer (BC), lung cancer (LC), esophageal cancer (EsC), gastric cancer (GC), bladder cancer (BLC), endometrial cancer (EC), ovarian cancer (OVC), head and neck cancer (HNC), and hematological malignancies indefinitely. Human PTK7 polypeptides include, but are not limited to, those having the amino acid sequence shown in UniProt identifier Q13308, and, but are not limited to, those having the amino acid sequences shown in Genbank, such as GenBank accession numbers NP_002812.2, NP_690619.1, NP_690620.1, NP_690621.1, NP_001257327.1, XP_011513067.1, XP_011513068.1, XP_047275113.1, XP_054212039.1, XP_054212040.1, and XP_054212041.1, which are incorporated herein by reference. Although lacking detectable catalytic tyrosine kinase activity, PTK7 overexpression is closely involved in Wnt signaling and promotes stem cell properties, survival, and tumor growth in cancer cells (see, for example, Atasaven et al., 2013; Cui et al., 2021; Gartner, 2014; Chen et al., 2014; Jiang et al., 2020; Shin et al., 2013; Liu et al., 2017; Lin et al., 2012; Ozcelik et al., 2020; Xiang et al., 2022; Wang et al., 2014; Prebet et al., 2010; Jiang et al., 2012; Damelin et al., 2017). PTK7 expression is generally low in normal tissues, but some protein expression has been reported in the bladder, kidneys, mammary glands, lungs, ovaries, uterus, and gastrointestinal tract, including dendritic cells (Damelin et al., 2017).Accordingly, PTK7 represents a promising target for novel anti-cancer therapies.

[0039] As used herein, "epitope" refers to an amino acid that is typically bound by an immunoglobulin VH / VL pair, such as the antibodies, antigen-binding portions thereof and other binding agents described herein. An epitope can be formed on a polypeptide from consecutive amino acids or non-consecutive amino acids aligned by the tertiary folding of a protein. Epitopes formed from consecutive amino acids are typically retained upon exposure to denaturing solvents, while epitopes formed by tertiary folding are typically impaired by treatment with denaturing solvents. An epitope typically comprises at least 3, more usually at least 5, about 9, or about 8 to 10 amino acids in a unique spatial arrangement. An epitope defines the minimal binding site for an antibody, its antigen-binding site and other binding agents, and thus represents the target of specificity of the antibody, its antigen-binding site or other immunoglobulin-based binding agents. In the case of a single domain antibody, an epitope represents the structural unit bound by the variable domain alone.

[0040] As used herein, "specifically binds" means that a binding agent (e.g., an antibody or antigen-binding portion thereof) described herein has a 10 -5 M (10000 nM) or less, for example, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12This refers to the ability to bind to a target such as human PTK7 with a KD of M or less. Specific binding can be influenced, for example, by the affinity and binding strength of the antibody, antigen-binding moiety, or other binding agent, as well as the concentration of the target polypeptide. Those skilled in the art can determine the appropriate conditions for the selective binding of the antibodies, antigen-binding moieties, and other binding agents described herein to PTK7 using any suitable method, such as titration of the antibody or other binding agent in a suitable cell-binding assay. A binding agent specifically bound to PTK7 is not substituted by dissimilar competing agents. In certain embodiments, a PTK7 antibody or its antigen-binding moiety or other binding agent is said to specifically bind to PTK7 when it preferentially recognizes its target antigen, PTK7, in a complex mixture of proteins and / or macromolecules.

[0041] In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is 10 -5 M (10000 nM) or less, for example, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of M or less. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is approximately 10 -5 M~10 -6 It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of M. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is approximately 10 -7 M~10 -8 It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of M. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is approximately 10 -8 M~10 -9It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of M. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is approximately 10 -9 M~10 -10 It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of M. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is approximately 10 -10 M~10 -11 It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of M. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is approximately 10 -11 M~10 -12 It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of M. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is approximately 10 -12 M~10 -13 It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of M. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance described herein is 10 -9 It specifically binds to the PTK7 polypeptide with a dissociation constant (KD) of less than M.

[0042] Unless otherwise specified, the term “alkyl” refers, by itself or as part of another term, to a substituted or unsubstituted linear or branched saturated hydrocarbon having the indicated number of carbon atoms (e.g., “-C1~C5 alkyl”, “-C1~C8 alkyl”, or “-C1~C 10(Alkyl refers to alkyl groups having 1-5, 1-8, or 1-10 carbon atoms, respectively.) Examples include methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), and 2-butyl (s-Bu, s-butyl, -CH(CH3)C H2CH3), 2-methyl-2-propyl(t-Bu, t-butyl, -C(CH3)3), 1-pentyl(n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl(-CH(CH3)CH2CH2CH3), 3-pentyl(--CH(CH2CH3)2), 2-methyl-2-butyl(-C(CH3)2CH2CH3), 3-methyl-2-butyl(-CH(CH3)CH(CH3)2), 3-methyl-1-butyl butyl(-CH2CH2CH(CH3)2), 2-methyl-1-butyl(-CH2CH(CH3)CH2CH3), 1-hexyl(-CH2CH2CH2CH2CH2CH3), 2-hexyl(-CH(CH3)CH2CH2CH2CH3), 3-hexyl(-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl(-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl(-CH( It contains CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl(-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl(-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl(-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl(-C(CH3)2CH(CH3)2), and 3,3-dimethyl-2-butyl(-CH(CH3)C(CH3)3).

[0043] Unless otherwise specified, the term “alkenyl” means, by itself or as part of another term, at least one unsaturated site (i.e., carbon-carbon sp). 2This refers to C2-C8 substituted or unsubstituted straight-chain or branched hydrocarbons having a double bond. Examples include, but are not limited to, ethylene or vinyl (-CH=CH2), allyl (-CH2CH=CH2), cyclopentenyl (-C5H7), and 5-hexenyl (-CH2CH2CH2CH2CH=CH2).

[0044] Unless otherwise specified, “alkynyl” refers, by itself or as part of another term, to a C2-C8 substituted or unsubstituted straight-chain or branched hydrocarbon having at least one unsaturated site (i.e., a carbon-carbon sp triple bond). Examples include, but are not limited to, acetylene and propargyl.

[0045] Unless otherwise specified, "alkylene" refers to a saturated, branched or linear hydrocarbon radical with 1 to 8 carbon atoms, having two monovalent radical centers induced by the removal of two hydrogen atoms from the same or two different carbon atoms of the parent alkane. Typical alkylene radicals include, but are not limited to, methylene (-CH2-), 1,2-ethyl (-CH2CH2-), 1,3-propyl (-CH2CH2CH2-), and 1,4-butyl (-CH2CH2CH2CH2-).

[0046] Unless otherwise specified, "alkenylene" refers to an unsaturated, branched or linear hydrocarbon radical having 2 to 8 carbon atoms and two monovalent radical centers induced by the removal of two hydrogen atoms from the same or two different carbon atoms of the parent alkene. Typical alkenylene radicals include 1,2-ethylene (-CH=CH-) indefinitely.

[0047] Unless otherwise specified, "alkynylene" refers to an unsaturated, branched, linear, or cyclic hydrocarbon radical having 2 to 8 carbon atoms and two monovalent radical centers induced by the removal of two hydrogen atoms from the same or two different carbon atoms of the parent alkyne. Typical alkynylene radicals include, but are not limited to, acetylene, propargyl, and 4-pentinyl.

[0048] Unless otherwise specified, the term “heteroalkyl” means, by itself or as part of another term, a substituted or unsubstituted stable linear or branched hydrocarbon, or a combination thereof, which is saturated and has 1 to 10, preferably 1 to 3, heteroatoms selected from the group consisting of O, N, Si, and S, where the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may be quaternized. The heteroatoms O, N, and S may be located at any internal position of the heteroalkyl group (i.e., as part of the main chain) or at a position where the alkyl group is bonded to the rest of the molecule. The heteroatom Si may be located at any position of the heteroalkyl group, including a position where the alkyl group is bonded to the rest of the molecule. Examples of heteroalkyl groups include: -CH2CH2OCH3, -CH2CH2NHCH3, -CH2CH2N(CH3)CH3, -CH2SCH2CH3, CH2CH2S(O)CH3, -CH2CH2S(O)2CH3, and -Si(CH3)3,-. For example, there may be up to two consecutive heteroatoms, such as -CH2NHOCH3 and CH2OSi(CH3)3. In some embodiments, the C1-C4 heteroalkyl group has 1-4 carbon atoms and 1 or 2 heteroatoms, and the C1-C3 heteroalkyl group has 1-3 carbon atoms and 1 or 2 heteroatoms.

[0049] Unless otherwise specified, the terms “heteroalkenyl” and “heteroalkynyl” refer, by themselves or as part of other terms, to a substituted or unsubstituted stable linear or branched alkenyl or alkynyl having 1 to 10, preferably 1 to 3, heteroatoms selected from the group consisting of O, N, Si, and S, where the nitrogen and sulfur atoms may be oxidized, and the nitrogen heteroatom may be quaternized. The heteroatoms O, N, and S may be located at any internal position of the heteroalkenyl or heteroalkynyl group (i.e., as part of the main chain) or at positions where alkyl groups are bonded to the rest of the molecule. The heteroatom Si may be located at any internal position of the heteroalkenyl or heteroalkynyl group, including positions where alkyl groups are bonded to the rest of the molecule.

[0050] Unless otherwise specified, the term “heteroalkylene” refers, either by itself or as part of another term, to substituted or unsubstituted divalent groups derived from heteroalkyls (discussed above), such as those exemplified by -CH2CH2SCH2CH2- and -CH2SCH2CH2NHCH2-. In some embodiments, C1-C4 heteroalkylenes have 1-4 carbon atoms and 1 or 2 heteroatoms, and C1-C3 heteroalkylenes have 1-3 carbon atoms and 1 or 2 heteroatoms. For heteroalkylene groups, the heteroatoms may occupy either or both of the chain ends. Furthermore, for alkylene and heteroalkylene linking groups, the orientation of the linking group is not implied.

[0051] Unless otherwise specified, the terms “heteroalkenylene” and “heteroalkynylene” refer, either by themselves or as part of other terms, to substituted or unsubstituted divalent groups derived from heteroalkenyl or heteroalkynyl (discussed above). In some embodiments, C2-C4 heteroalkenylenes or heteroalkynylenes have 1-4 carbon atoms. For heteroalkenylene and heteroalkynylene groups, the heteroatom may occupy either or both of the chain ends. Furthermore, for heteroalkenylene and heteroalkynylene linking groups, the orientation of the linking group is not implied.

[0052] Unless otherwise specified, “C3-C8 carbocyclic ring” refers, by itself or as part of another term, to a substituted or unsubstituted 3, 4, 5, 6, 7, or 8-membered monovalent, substituted or unsubstituted, saturated or unsaturated non-aromatic monocyclic or bicyclic carbocyclic ring, which is derived by the removal of one hydrogen atom from the ring atom of the parent ring system. Representative C3-C8 carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, cycloheptyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptatrielinyl, cyclooctyl, and cyclooctadienyl.

[0053] Unless otherwise specified, "C3-C8 carbocyclo" refers, by itself or as part of another term, to the substituted or unsubstituted C3-C8 carbocyclic group as defined above, where another hydrogen atom of the carbocyclic group is substituted by a bond (i.e., divalent).

[0054] Unless otherwise specified, "C3~C 10 A "carbocyclic ring" refers, by itself or as part of other terms, to a substituted or unsubstituted 3, 4, 5, 6, 7, 8, 9, or 10-membered monovalent, substituted or unsubstituted, saturated or unsaturated non-aromatic monocyclic, bicyclic, or tricyclic carbocyclic ring, which is induced by the removal of one hydrogen atom from the ring atom of the parent ring system. A typical example is -C3~C 10The carbocyclic ring includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, cycloheptyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptatrielinyl, cyclooctyl, and cyclooctadienyl. -C3~C 10 The carbocyclic rings may further include condensed cyclooctinate carbocyclic rings such as condensed cyclooctinate compounds disclosed in International Publication No. WO2011 / 136645 (the disclosure of which is incorporated herein by reference), including BCN (bicyclo[6.1.0]nonine) and DBCO (dibenzocyclooctin).

[0055] Unless otherwise specified, “C3-C8 heterocycle” refers, either by itself or as part of another term, to a substituted or unsubstituted, monovalent aromatic or non-aromatic monocyclic or bicyclic ring system having 3 to 8 carbon atoms (also called ring members) and 1 to 4 heteroatoms independently selected from N, O, P, or S, induced by the removal of one hydrogen atom from the ring atoms of the parent ring system. One or more N, C, or S atoms in the heterocycle may be oxidized. The ring containing the heteroatoms may be aromatic or non-aromatic. Unless otherwise specified, the heterocycle is bonded to its pendant group at any heteroatom or carbon atom that results in a stable structure. Representative examples of C3-C8 heterocycles include, but are not limited to, pyrrolidinyl, azetidinyl, piperidinyl, morpholinyl, tetrahydrofuranil, tetrahydropyranil, benzofuranil, benzothiophene, indolyl, benzopyrazolyl, pyrrolyl, thiophenyl (thiophene), furanil, thiazolyl, imidazolyl, pyrazolyl, pyrimidinyl, pyridinyl, pyrazinyl, pyridadinyl, isothiazolyl, and isoxazolyl. Unless otherwise specified, the term "heterocarbocyclic" is synonymous with the terms "heterocycle" or "heterocyclo" as used herein.

[0056] Unless otherwise specified, "C3-C8 heterocyclo" refers, by itself or as part of other terms, to the substituted or unsubstituted C3-C8 heterocyclic group as defined above, where one of the hydrogen atoms of the carbocyclic group is substituted by a bond (i.e., divalent).

[0057] Unless otherwise specified, “aryl” means, by itself or as part of another term, a substituted or unsubstituted monovalent carbocyclic aromatic hydrocarbon radical with 6 to 20 carbon atoms (preferably 6 to 14 carbon atoms) derived by the removal of one hydrogen atom from one carbon atom of an aromatic ring system. Some aryl groups are represented as “Ar” in exemplary structures. Typical aryl groups include, non-limitingly, radicals derived from benzene, substituted benzenes, naphthalenes, anthracenes, biphenyls, and others. An exemplary aryl group is the phenyl group.

[0058] Unless otherwise specified, “arylene” means, by itself or as part of another term, an unsubstituted or substituted aryl group as defined above, where one of the hydrogen atoms of the aryl group is substituted by a bond (i.e., divalent) and may be ortho, meta, or para oriented.

[0059] Unless otherwise specified, “heteroaryl” and “heterocyclic” refer to a ring system in which one or more ring atoms are heteroatoms, such as nitrogen, oxygen, and sulfur. Heterocyclic radicals contain 1 to 20 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S. Heterocyclics may be monocyclic with 3 to 7 ring members (2 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S) or dicyclic with 7 to 10 ring members (4 to 9 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S), such as bicyclo[4,5], [5,5], [5,6], or [6,6] systems.

[0060] Unless otherwise specified, “heteroarylene” means, by itself or as part of another term, an unsubstituted or substituted heteroaryl group as defined above, where one of the hydrogen atoms of the heteroaryl group is substituted by a bond (i.e., divalent).

[0061] Unless otherwise specified, "carboxyl" is COOH or COO - M + It refers to M + It is a cation.

[0062] Unless otherwise specified, "oxo" refers to (C=O).

[0063] Unless otherwise specified, "substituted alkyl" and "substituted aryl" refer to alkyl and aryl compounds, respectively, in which one or more hydrogen atoms are independently substituted with substituents. Typical substituents are -X and -R. 10 , -O - , -OR 10 , -SR 10 , -S - , -NR 10 2. -NR 10 3, =NR 10 , -CX3, -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO2, =N2, -N3, -NR 10 C(=O)R 10 -C(=O)R 10 -C(=O)NR 10 2, -SO3 - -SO3H, -S(=O)2R 10 -OS(=O)2OR 10 -S(=O)2NR 10 -S(=O)R 10 , -OP(=O)(OR 10 )2, -P(=O)(OR 10 )2, -PO - 3, -PO3H2, -AsO2H2, -C(=O)R 10 -C(=O)X, -C(=S)R 10 , -CO2R 10 , -CO2 - , -C(=S)OR10 , C(=O)SR 10 , C(=S)SR 10 , C(=O)NR 10 2. C(=S)NR 10 2, or C(=NR 10 )NR 10 2 non-restrictively includes, where each X is independently a halogen: -F, -Cl, -Br, or -I; each R 10 These are independently -H, -C1~C 20 Alkyl, -C6~C 20 Ariel, -C3~C 14 These are heterocyclic, protecting groups, or prodrug moieties. Typical substituents include (=O). The alkylene, carbocyclic, carbocyclo, arylene, heteroalkyl, heteroalkylene, heterocyclic, and heterocyclo groups described above may also be substituted in the same way.

[0064] Unless otherwise specified, “polyhydroxyl group” refers to an alkyl, alkylene, carbocyclic, or carbocyclo group in which two or more hydrogen atoms on the carbon atoms of the carbon chain are substituted with hydroxyl groups, or three or more hydroxyl groups. In some embodiments, a polyhydroxyl group contains at least three hydroxyl groups. In some embodiments, a polyhydroxyl group contains carbon atoms with exactly one hydroxyl group per carbon atom. A polyhydroxyl group may contain one or more carbon atoms that are not substituted with hydroxyl groups. Each carbon atom of a polyhydroxyl group may be substituted with a hydroxyl group. Examples of polyhydroxyl groups include linear (acyclic) or cyclic monosaccharides such as C6 or C5 sugars like glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, growth, idose, talose, aldose, and ketose; sugar acids such as gluconic acid, aldonic acid, uronic acid, or urosonic acid; and amino sugars such as glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine. In some embodiments, polyhydroxyl groups include linear or cyclic disaccharides and polysaccharides.

[0065] Unless otherwise specified by context, “may be substituted” means alkyl, alkenyl, alkynyl, alkylaryl, arylalkyl heterocycle, aryl, heteroaryl, alkylheteroaryl, heteroarylalkyl, or other substituents, moieties, or groups as defined or disclosed herein, wherein the hydrogen atoms of the substituents, moieties, or groups may be substituted with different moieties or groups, or an alicyclic carbon chain containing one of the substituents, moieties, or groups is interrupted by substituting a carbon atom in the chain with a different moiety or group. In some aspects, an alkene functional group substitutes two consecutive sp3 carbon atoms of an alkyl substituent, but the radical carbon of the alkyl moiety is not substituted, so the alkyl that may be substituted is an unsaturated alkyl substituent.

[0066] Any optional substituent replacing hydrogen in any of the aforementioned substituents, moieties or groups is independently selected from the group consisting of aryl, heteroaryl, hydroxyl, alkoxy, aryloxy, cyano, halogen, nitro, fluoroalkoxy, and amino including mono-, di- and tri-substituted amino groups, and protected derivatives thereof, or selected from the group consisting of -X, -OR', -SR', -NH₂, -N(R')(R"), -N(R")₃, =NR, -CX₃, -CN, -NO₂, -NR'C(=O)H, -NR'C(=O)R, -NR'C(=O)R", -C(=O)R', -C(=O)NH₂, -C(=O)N(R')R", -S(=O)₂R", -S(=O)₂NH₂, -S(=O)₂N(R')R", -S(=O)₂NH₂, -S(=O)₂N(R')R", -S(=O)₂OR', -S(=O)R", -OP(=O)(OR')(OR"), -OP(OH)₃, -P(=O)(OR')(OR"), -PO₃H₂, -C(=O)R', -C(=S)R", -CO₂R', -C(=S)OR", -C(=O)SR', -C(=S)SR', -C(=S)NH₂, -C(=S)N(R')(R")₂, -C(=NR')NH₂, -C(=NR')N(R')R", and salts thereof, wherein each X is independently selected from the group consisting of halogen: -F, -Cl, -Br, and -I; wherein each R" is independently C1 to C 20 alkyl, C2 to C 20 alkenyl, C2 to C 20 alkynyl, C6 to C 24 aryl, C3 to C 24 heterocyclyl (including C5 to C 24 heteroaryl), a protecting group, and a prodrug moiety, or two of R" together with the heteroatom to which they are attached define a heterocyclyl; R' is hydrogen or R", wherein R" is C1 to C 20 alkyl, C6 to C 24 aryl, C3 to C 24 heterocyclyl (including C5 to C 24 heteroaryl), and a protecting group.

[0067] Typically, any optional substituent is selected from the group consisting of -X, -OH, -OR'', -SH, -SR'', -NH2, -NH(R''), -NR'(R'')2, -N(R'')3, =NH, =NR'', -CX3, -CN, -NO2, -NR'C(=O)H, NR'C(=O)R'', -CO2H, -C(=O)H, -C(=O)R'', -C(=O)NH2, -C(=O)NR'R'' - -S(=O)2R'', -S(=O)2NH2, -S(=O)2N(R')R'', -S(=O)2NH2, -S(=O)2N(R')(R''), -S(=O)2OR', -S(=O)R'', -C(=S)R'', -C(=S)NH2, -C(=S)N(R')R'', -C(=NR')N(R'')2, and salts thereof, wherein each X is independently selected from the group consisting of -F and -Cl, and R'' is typically C1-C6 alkyl, C6-C 10 aryl, C3-C 10 heterocyclyl (including C5-C 10 heteroaryl), and a protecting group, selected from the group consisting of; R' is independently hydrogen, C1-C6 alkyl independently selected from R'', C6-C 10 aryl, C3-C 10 heterocyclyl (including C5-C 10 heteroaryl), and a protecting group. More typically, the substituent is selected from the group consisting of -X, -R'', -OH, -OR'', -NH2, -NH(R''), -N(R'')2, -N(R'')3, -CX3, -NO2, -NHC(=O)H, -NHC(=O)R'', -C(=O)NH2, -C(=O)NHR'', -C(=O)N(R'')2, -CO2H, -CO2R'', -C(=O)H, -C(=O)R'', -C(=O)NH2, -C(=O)NH(R''), -C(=O)N(R'')2, -C(=NR')NH2, -C(=NR')NH(R''), -C(=NR')N(R'')2, a protecting group and salts thereof, wherein each X is -F, and R'' is independently C1-C6 alkyl, C6-C 10 aryl, C5-C 10 selected from the group consisting of heteroaryl and a protecting group; R' is selected from the group consisting of hydrogen, C1-C6 alkyl independently selected from R'', and a protecting group.

[0068] The compounds of the present invention or their pharmaceutically acceptable salts may contain one or more asymmetric centers, thus giving rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined from an absolute stereochemical standpoint as (R) or (S) or, for amino acids, (D) or (L). The present invention means that it includes all such possible isomers as well as their racemic and optically pure forms. The optically active (+) and (-), (R) and (S), or (D) and (L) isomers may be prepared using chiral synthons or chiral reagents, or separated using conventional techniques, e.g., chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors or separation of racemates (or racemates of salts or derivatives) using, for example, chiral high-pressure liquid chromatography (HPLC). Where the compounds described herein contain geometrically asymmetric olefin double bonds or other centers, unless otherwise specified, the compounds are intended to include both E and Z geometric isomers. Similarly, it is intended that all tautomer forms be included.

[0069] A "stereoisomer" refers to a compound that is composed of the same atoms bonded together by the same bonds, but has different, incompatible three-dimensional structures. This invention envisions various stereoisomers and mixtures thereof, including enantiomers, which refer to two stereoisomers that are mirror images of each other and whose molecules cannot be superimposed. This invention also includes "diastereomers," which refer to two or more stereoisomers of a compound that have different configurations in one or more equivalent stereocenters and are not mirror images of each other.

[0070] Throughout this specification, the structures are depicted with specific stereocenters, but this specification should be read to include variations of those stereocenters. For example, the structure of exatecan may be shown in the (S,S) configuration, but it is also anticipated that in other embodiments of the complex described herein, a (R,S) diastereomer of exatecan may be found.

[0071] Unless otherwise specified, “drug unit” or “drug” refers to cytotoxic substances (such as chemotherapeutic agents or drugs), immunomodulatory substances, nucleic acids (including siRNA), growth inhibitors, toxins (e.g., prototoxins, bacterial, fungal, plant or animal-derived enzymatic toxins, or fragments thereof), radioisotopes, PROTACs, and other compounds that become active against target cells when delivered to them.

[0072] Unless otherwise specified, the term “polymer unit” refers to a polymer portion consisting of repeating subunits. Examples of polymer units include polyamides and polyethers. In some embodiments, the polymer unit is selected from substituted polyamides, substituted polyethers, or combinations thereof. In further embodiments, the polymer unit is, (i) Each R a H or C 1~6 It is alkyl, and each R b H or C 1~6 It is alkyl, n 0 The equation is such that the numbers are independently 2 to 26: substituted polyamides or stereoisomers thereof containing TIFF2026534278000002.tif18128; (ii) Each R b H or C 1~6 It is alkyl, n 0 The equation is such that the numbers are independently 2 to 26: Substituted polyethers containing TIFF2026534278000003.tif20128 or their stereoisomers; or (iii) combinations of those More likely to be selected.

[0073] Unless otherwise specified, the terms "sugar unit" or "sugar group" refer to a carbohydrate group. Examples of sugar units include glycosides.

[0074] Unless otherwise specified, the terms “carboxyl unit” or “carboxyl group” refer to groups containing a carbonyl group [-C(O)-], a carboxyl group [-CO2H], and / or a carboxylate group [-CO2M] where M refers to a cationic counterion.

[0075] Unless otherwise specified, the term "stretcher group" refers to the linking portion that connects the PTK7 binding material to a group that can be enzymatically cleaved.

[0076] Unless otherwise specified, the term "polyamide" refers to a polymer group consisting of repeating subunits containing amide bonds.

[0077] Unless otherwise specified, the term "polyether" refers to a polymer group consisting of repeating subunits containing ether bonds.

[0078] Unless otherwise specified, the term “enzyme-cleavable group” refers to a group that becomes cleavable by metabolic processes or reactions in the intracellular or extracellular environment, thereby cleaving the covalent bond between the drug unit (e.g., a cytotoxic substance) and the linker unit or a portion thereof, resulting in the release of the free drug unit or the linker unit-drug metabolite dissociated from the rest of the linker unit.

[0079] As used herein, the term “pharmaceutically acceptable salt” refers to a pharmaceutically acceptable organic or inorganic salt of a compound (e.g., a linker, drug linker, or complex). The compound typically contains at least one amino group and can therefore form an acid addition salt with this amino group. Exemplary salts include, but are not limited to, sulfates, citrates, acetates, oxalates, chlorides, bromides, iodides, nitrates, bisulfates, phosphates, acidic phosphates, isonicotinates, lactates, salicylates, acidic citrates, tartrates, oleates, tannates, pantothenates, bitartrates, ascorbicates, succinates, lineates, gentisinates, fumarates, glucons, glucurons, sugarates, formates, benzoates, glutamates, methanesulfons, ethanesulfons, benzenesulfons, toluenesulfons, and pamoates (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Pharmacochemically acceptable salts may involve the inclusion of other molecules such as acetate ions, succinate ions, or other counterions. The counterion may be any organic or inorganic moiety that stabilizes the charge of the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. If multiple charged atoms are part of a pharmaceutically acceptable salt, multiple counterions may be present. Therefore, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counterions.

[0080] As used herein, the term “essentially derived from” refers to elements necessary for a given embodiment. The term permits the presence of elements that do not substantially affect the basic, novel, or functional characteristics of that embodiment.

[0081] As used herein, the term "consisting of" refers to the compositions, methods, and their respective components described herein, excluding any elements not described in the description of the embodiments.

[0082] Except as an example, or unless otherwise indicated, all numerical values ​​used herein to represent quantities of components or reaction conditions should be understood to be modified in all cases by the term “approximately.” When used in relation to percentages, the term “approximately” may mean + / - 1%.

[0083] The terms "statistically significant" or "significant" refer to statistical significance, generally meaning a difference of two standard deviations (2SD) above or below the baseline.

[0084] Other terms are defined herein within the descriptions of various aspects of the invention.

[0085] Antibodies and binding agents PTK7-conjugated antibodies (also called PTK7 antibodies) and their antigen-binding moieties, as well as other conjugates that specifically bind to human PTK7, are provided herein. Complexes of PTK7 antibodies, antigen-binding moieties, and other conjugates (also called PTK7 complexes) conjugated to drugs such as cytotoxic substances or immunomodulators are also provided herein. In some embodiments, the PTK7 antibody, antigen-binding moiety, other conjugates, and / or PTK7 complex specifically bind to and reduce the number of PTK7+ cells in a subject. In some embodiments, the PTK7 antibody, antigen-binding moiety, other conjugates, and / or PTK7 complex specifically bind to and reduce the number of PTK7+ cancer cells in a subject. In some embodiments, the PTK7 antibody, antigen-binding moiety, other conjugates, and / or PTK7 complex specifically bind to and reduce the number of PTK7+ cells associated with a disease or condition such as cancer or autoimmune disease in a subject. In some embodiments, a PTK7 antibody, antigen-binding moiety, other binding agents, and / or PTK7 complex specifically bind to and reduce the number of PTK7+ cells associated with a disease or condition in a subject such as a human or animal.

[0086] In some embodiments, the PTK7 antibody or its antigen-binding moiety comprises a heavy-chain variable (VH) region and a light-chain variable (VL) region, and the VH and VL regions have amino acid sequences shown in pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, SEQ ID NO:15 and SEQ ID NO:16, SEQ ID NO:34 and SEQ ID NO:35, SEQ ID NO:41 and SEQ ID NO:42, and SEQ ID NO:48 and SEQ ID NO:49, respectively. In some embodiments, the PTK7 antibody or its antigen-binding moiety comprises a heavy-chain variable (VH) region and a light-chain variable (VL) region, and the VH and VL regions have amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2, respectively. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, and the VH and VL regions have amino acid sequences shown in SEQ ID NO: 8 and SEQ ID NO: 9, respectively. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, and the VH and VL regions have amino acid sequences shown in SEQ ID NO: 15 and SEQ ID NO: 16, respectively. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, and the VH and VL regions have amino acid sequences shown in SEQ ID NO: 34 and SEQ ID NO: 35, respectively. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, and the VH and VL regions have amino acid sequences shown in SEQ ID NO: 41 and SEQ ID NO: 42, respectively. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:48 and SEQ ID NO:49, respectively.

[0087] In some embodiments, the PTK7 antibody or its antigen-binding moiety comprises a heavy-chain variable (VH) region and a light-chain variable (VL) region, the VH and VL regions having amino acid sequences represented by pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, SEQ ID NO:15 and SEQ ID NO:16, SEQ ID NO:34 and SEQ ID NO:35, SEQ ID NO:41 and SEQ ID NO:42, and SEQ ID NO:48 and SEQ ID NO:49, respectively, wherein the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, the PTK7 antibody or its antigen-binding moiety comprises a heavy-chain variable (VH) region and a light-chain variable (VL) region, the VH and VL regions having amino acid sequences represented by pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, SEQ ID NO:15 and SEQ ID NO:16, SEQ ID NO:34 and SEQ ID NO:35, SEQ ID NO:41 and SEQ ID NO:42, and SEQ ID NO:48 and SEQ ID NO:49, respectively, wherein the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, wherein the CDR of the heavy-chain or light-chain variable region is not modified. The expression "wherein the CDR of the heavy-chain or light-chain variable region is not modified" refers to the CDR of the VH and VL regions that have no amino acid substitutions, deletions, or insertions.

[0088] In some embodiments, the PTK7 antibody or its antigen-binding moiety comprises a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences that have at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% similarity to pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, SEQ ID NO:15 and SEQ ID NO:16, SEQ ID NO:34 and SEQ ID NO:35, SEQ ID NO:41 and SEQ ID NO:42, and SEQ ID NO:48 and SEQ ID NO:49, respectively. The levels of similarity are due to amino acid substitutions, deletions, or insertions in the VH and VL sequences shown in SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, SEQ ID NO:15 and SEQ ID NO:16, SEQ ID NO:34 and SEQ ID NO:35, SEQ ID NO:41 and SEQ ID NO:42, and SEQ ID NO:48 and SEQ ID NO:49, respectively.

[0089] In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified.

[0090] In some embodiments, the PTK7 antibody or its antigen-binding portion comprises a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the amino acid sequence shown in SEQ ID NO:1 and SEQ ID NO:2, respectively.

[0091] In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:8 and SEQ ID NO:9, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4 or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:8 and SEQ ID NO:9, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4 or 1-2 amino acid substitutions, deletions or insertions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified.

[0092] In some embodiments, the PTK7 antibody or its antigen-binding portion comprises a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the amino acid sequence shown in SEQ ID NO:8 and SEQ ID NO:9, respectively.

[0093] In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:15 and SEQ ID NO:16, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4 or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:15 and SEQ ID NO:16, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4 or 1-2 amino acid substitutions, deletions or insertions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified.

[0094] In some embodiments, the PTK7 antibody or its antigen-binding portion comprises a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the amino acid sequence shown in SEQ ID NO:15 and SEQ ID NO:16, respectively.

[0095] In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:34 and SEQ ID NO:35, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified.

[0096] In some embodiments, the PTK7 antibody or its antigen-binding portion comprises a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the amino acid sequence shown in SEQ ID NO:34 and SEQ ID NO:35, respectively.

[0097] In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:41 and SEQ ID NO:42, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified. In some embodiments, the PTK7 antibody or its antigen-binding portion includes a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:41 and SEQ ID NO:42, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified.

[0098] In some embodiments, the PTK7 antibody or its antigen-binding portion comprises a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the amino acid sequence shown in SEQ ID NO:41 and SEQ ID NO:42, respectively.

[0099] In some embodiments, the PTK7 antibody or its antigen-binding moiety includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:48 and SEQ ID NO:49, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, the PTK7 antibody or its antigen-binding moiety includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, the VH and VL regions having amino acid sequences shown in SEQ ID NO:48 and SEQ ID NO:49, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, where the CDR of the heavy-chain or light-chain variable region is not modified.

[0100] In some embodiments, the PTK7 antibody or its antigen-binding portion comprises a heavy chain variable (VH) region and a light chain variable (VL) region, the VH and VL regions having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% of the amino acid sequence shown in SEQ ID NO:48 and SEQ ID NO:49, respectively.

[0101] In some embodiments, a binding agent is provided herein comprising a VH region and a VL region, wherein the heavy-chain variable (VH) and light-chain variable (VL) regions have amino acid sequences represented by pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, and SEQ ID NO:15 and SEQ ID NO:16, respectively, and the binding agent specifically binds to human PTK7. In some embodiments, the binding agent comprises a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences represented by pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, and SEQ ID NO:15 and SEQ ID NO:16, respectively, and the binding agent specifically binds to human PTK7 with a binding affinity (lower Kd) higher than or similar to that of the antibody cofetuzumab. In some embodiments, a conjugated material comprising VH and VL regions is provided herein, wherein the heavy chain variable region (VH) and the light chain variable region (VL) have amino acid sequences shown in pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, and SEQ ID NO:15 and SEQ ID NO:16, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified.In some embodiments, a conjugate is provided herein that includes a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH and VL regions have amino acid sequences represented by pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, and SEQ ID NO:15 and SEQ ID NO:16, respectively, where the heavy chain and light chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, where the CDR of the heavy chain or light chain variable region is not modified. As described herein, the conjugate contains a PTK7 antibody or its antigen-binding moiety, and may also contain other peptides or polypeptides covalently bound to the PTK7 antibody or its antigen-binding moiety. In any of these embodiments, the conjugate specifically binds to human PTK7.

[0102] In some embodiments, a binding agent is provided herein that includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2, respectively, and the binding agent specifically binds to human PTK7. In some embodiments, the binding agent includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2, respectively, and the binding agent specifically binds to human PTK7 with a binding affinity (lower Kd) that is higher than or similar to that of the antibody cofetuzumab. In some embodiments, a conjugating substance is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy-chain or light-chain variable region is not modified.

[0103] In some embodiments, a binding agent is provided herein that includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:8 and SEQ ID NO:9, respectively, and the binding agent specifically binds to human PTK7. In some embodiments, the binding agent includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:8 and SEQ ID NO:9, respectively, and the binding agent specifically binds to PTK7 with a higher binding affinity (lower Kd) than the antibody cofetuzumab. In some embodiments, a conjugating substance is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:8 and SEQ ID NO:9, respectively, where the heavy-chain and light-chain variable framework regions may be modified in the framework region by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions, where the CDR of the heavy-chain or light-chain variable region is not modified.

[0104] In some embodiments, a binding agent is provided herein that includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:15 and SEQ ID NO:16, respectively, and the binding agent specifically binds to human PTK7. In some embodiments, the binding agent includes a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:15 and SEQ ID NO:16, respectively, and the binding agent specifically binds to PTK7 with a higher binding affinity (lower Kd) than the antibody cofetuzumab. In some embodiments, a conjugating substance is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:15 and SEQ ID NO:16, respectively, where the heavy-chain and light-chain variable framework regions may be modified in the framework region by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions, where the CDR of the heavy-chain or light-chain variable region is not modified.

[0105] In some embodiments, an antibody or antigen-binding moiety is provided comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 arranged in the heavy chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 arranged in the light chain variable region framework region, and the CDRs of VH and VL are (i) SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, FAS, and SEQ ID NO:7, respectively; (ii) SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, RTS, and SEQ ID NO:15, respectively; (iii) SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, KVS, and SEQ ID NO:22, respectively; (iv) SEQ ID (v) SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, KVS, and SEQ ID NO:27 respectively, (v) SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33 respectively, (vi) SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, FAS, and SEQ ID NO:40 respectively, and (vii) SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, RTS, and SEQ ID NO:47 respectively. In some embodiments, each VH and VL region includes a humanized framework region. In some embodiments, each VH and VL region includes a humanized framework region.

[0106] In some embodiments, an antibody or antigen-binding moiety is provided, comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within the heavy chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within the light chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, FAS, and SEQ ID NO:7, respectively. In some embodiments, each VH and VL region comprises a humanized framework region. In some embodiments, each VH and VL region comprises a humanized framework region.

[0107] In some embodiments, an antibody or antigen-binding moiety is provided, comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within a heavy chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within a light chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, RTS, and SEQ ID NO:14, respectively. In some embodiments, each VH and VL region comprises a humanized framework region. In some embodiments, each VH and VL region comprises a humanized framework region.

[0108] In some embodiments, an antibody or antigen-binding moiety is provided, comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within a heavy chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within a light chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, KVS, and SEQ ID NO:22, respectively. In some embodiments, each VH and VL region comprises a humanized framework region. In some embodiments, each VH and VL region comprises a humanized framework region.

[0109] In some embodiments, an antibody or antigen-binding moiety is provided, comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within a heavy chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within a light chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, KVS, and SEQ ID NO:27, respectively. In some embodiments, each VH and VL region comprises a humanized framework region. In some embodiments, each VH and VL region comprises a humanized framework region.

[0110] In some embodiments, an antibody or antigen-binding moiety is provided, comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within a heavy chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within a light chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33, respectively. In some embodiments, each VH and VL region comprises a humanized framework region. In some embodiments, each VH and VL region comprises a humanized framework region.

[0111] In some embodiments, a conjugate is provided comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within the heavy-chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within the light-chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, FAS, and SEQ ID NO:7, respectively. In some embodiments, each VH and VL region comprises a humanization framework region. In some embodiments, each VH and VL region comprises a human framework region. In some embodiments, a binding material is provided comprising a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region comprises complementarity determination regions HCDR1, HCDR2, and HCDR3 arranged in the heavy chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 arranged in the light chain variable region framework region, and the CDRs of the VH and VL are (i) SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, FAS, and SEQ ID NO:7 respectively, (ii) SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, RTS, and SEQ ID NO:14 respectively, (iii) SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, KVS, and SEQ ID NO:22 respectively, (iv) SEQ ID The amino acid sequence is represented by a set of amino acid sequences selected from NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, KVS, and SEQ ID NO:27, as well as (v)SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33. In some embodiments, each VH and VL region includes a humanized framework region.In some embodiments, each VH and VL region includes a human framework region.

[0112] In some embodiments, a conjugate is provided comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within the heavy-chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within the light-chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, FAS, and SEQ ID NO:7, respectively. In some embodiments, each VH and VL region comprises a humanization framework region. In some embodiments, each VH and VL region comprises a human framework region.

[0113] In some embodiments, a conjugate is provided comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within the heavy-chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within the light-chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, RTS, and SEQ ID NO:14, respectively. In some embodiments, each VH and VL region comprises a humanization framework region. In some embodiments, each VH and VL region comprises a human framework region.

[0114] In some embodiments, a conjugate is provided comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within the heavy-chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within the light-chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, KVS, and SEQ ID NO:22, respectively. In some embodiments, each VH and VL region comprises a humanized framework region. In some embodiments, each VH and VL region comprises a humanized framework region.

[0115] In some embodiments, a conjugate is provided comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within the heavy-chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within the light-chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, KVS, and SEQ ID NO:27, respectively. In some embodiments, each VH and VL region comprises a humanization framework region. In some embodiments, each VH and VL region comprises a human framework region.

[0116] In some embodiments, a conjugate is provided comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within the heavy-chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within the light-chain variable region framework region, and the CDRs of VH and VL have amino acid sequences shown in SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33, respectively. In some embodiments, each VH and VL region comprises a humanization framework region. In some embodiments, each VH and VL region comprises a human framework region.

[0117] In some embodiments, a binding material is provided comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH region comprises complementarity-determining regions HCDR1, HCDR2, and HCDR3 located within the heavy-chain variable region framework region, and the VL region comprises LCDR1, LCDR2, and LCDR3 located within the light-chain variable region framework region, and the CDRs of the VH and VL are respectively determined according to the IMGT unique numbering scheme. It has an amino acid sequence selected from the group consisting of TIFF2026534278000004.tif38128. Lefranc, MP The Immunologist, 7, 132-136 (1999). In some embodiments, each VH and VL region includes a humanized framework region. In some embodiments, each VH and VL region includes a human framework region.

[0118] In some embodiments, a binding material is provided that includes a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region includes complementarity determination regions HCDR1, HCDR2, and HCDR3 located within the heavy chain variable region framework region, and the VL region includes LCDR1, LCDR2, and LCDR3 located within the light chain variable region framework region, and the CDRs of the VH and VL are respectively numbered according to the KABAT numbering scheme. It has an amino acid sequence selected from the group consisting of TIFF2026534278000005.tif38128. Dondelinger M. et al. Frontiers in Immunology, 9, 2278 (2018). In some embodiments, each VH and VL region includes a humanized framework region. In some embodiments, each VH and VL region includes a human framework region.

[0119] In some embodiments, a binding substance is provided comprising a heavy chain having a heavy chain variable (VH) region and a heavy chain constant region, and a light chain having a light chain variable (VL) region and a light chain constant region, wherein the VH region has the amino acid sequence shown in SEQ ID NO:15, the VL region has the amino acid sequence shown in SEQ ID NO:16, the heavy chain constant region has the amino acid sequence shown in SEQ ID NO:56, and the light chain constant region has the amino acid sequence shown in SEQ ID NO:57.

[0120] In some embodiments, the compositions and methods described herein relate to the reduction of PTK7+ cells in a subject in vivo by PTK7 antibodies, their antigen-binding moieties, other conjugates, or complexes thereof (e.g., reducing the number of PTK7+ cells in cancer or tumors, or the number of PTK7+ cells associated with autoimmune diseases or disorders). In some embodiments, the compositions and methods described herein relate to the treatment of PTK7+ cancer in a subject by administering PTK7 antibodies, their antigen-binding moieties, other conjugates, or complexes thereof. In some embodiments, the compositions and methods described herein relate to the treatment of autoimmune diseases in a subject by administering PTK7 antibodies, their antigen-binding moieties, other conjugates, or complexes thereof. In some embodiments, the compositions and methods described herein relate to the treatment of diseases or disorders associated with PTK7+ cells in a subject by administering PTK7 antibodies, their antigen-binding moieties, other conjugates, or complexes thereof. In any of these embodiments, the methods further include reducing the number of PTK7+ cells associated with a disease, condition, or cancer in a subject.

[0121] As used herein, the term “antibody” refers to an immunoglobulin molecule and a molecule containing an immunoactive portion of an immunoglobulin molecule, i.e., an antigen-binding site that specifically binds to an antigen, such as human PTK7. The term generally refers to an antibody containing two immunoglobulin heavy chain variable regions and two immunoglobulin light chain variable regions, including a full-length antibody (having heavy and light chain constant regions).

[0122] Each heavy chain consists of a variable region (abbreviated as VH) and a constant region. The heavy chain constant region may contain three domains CH1, CH2, and CH3, and optionally a fourth domain, CH4. Each light chain consists of a variable region (abbreviated as VL) and a constant region. The light chain constant region is the CL domain. The VH and VL regions may be further divided into a highly variable region called the complementarity-determining region (CDR) and scattered conserved regions called the framework region (FR). Thus, each VH and VL region consists of three CDRs and four FRs arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4 from the N-terminus to the C-terminus. This structure is well known to those skilled in the art.

[0123] As used herein, the “antigen-binding moiety” of a PTK7 antibody refers to the portion of the PTK7 antibody described herein that has the VH and VL sequences of the PTK7 antibody, or the CDR of the PTK7 antibody, and specifically binds to PTK7. Examples of antigen-binding moieties include Fab, Fab', F(ab')2, Fv, scFv, disulfide-bound Fv, single-domain antibodies (also called VHH, VNAR, sdAb, or nanobody), or diabodies (see, for example, Huston et al., Proc. Natl. Acad. Sci. USA, 85, 5879-5883 (1988) and Bird et al., Science 242, 423-426 (1988), which are incorporated herein by reference). As used herein, the terms Fab, F(ab')2, and Fv refer to: (i) a Fab fragment is a monovalent fragment consisting of VL, VH, CL, and CH1 domains; (ii) an F(ab')2 fragment is a bivalent fragment containing two Fab fragments linked to each other in a hinge region via disulfide crosslinks; and (iii) an Fv fragment consists of VL and VH domains, but in any case is a PTK7 antibody. The two domains of the Fv fragment, namely VL and VH, are encoded by separate coding regions, but they may be further linked to each other using a synthetic linker, e.g., a poly-G4S amino acid sequence (disclosed as SEQ ID NO: 58, '(G4S)n', where n=1 to 5), making it possible to prepare a single protein chain (known as single-chain Fv or scFv) in which the VL and VH regions combine to form a monovalent molecule. The term “antigen-binding site” of an antibody is also intended to include such single-chain antibodies. Other forms of single-chain antibodies, such as "diabody," are also included here.Diabody is a bivalent, bispecific antibody in which the VH and VL domains are expressed on a single polypeptide chain. However, the linker that binds the VH and VL domains is too short for the two domains to bind on the same chain. This causes the VH and VL domains to be paired with complementary domains (VL and VH, respectively) on different chains, forming two antigen-binding sites (see, for example, Holliger, R, et al. (1993) Proc. Natl. Acad. Sci. USA 90:64446448; Poljak, R. J, et al. (1994) Structure 2:1121-1123).

[0124] A single-domain antibody is an antibody moiety consisting of a single monomeric variable antibody domain. Single-domain antibodies can be derived from variable domains (e.g., nanobody or VHH moieties) of antibody heavy chains derived from camelid animals. Furthermore, the term single-domain antibody includes autonomous human heavy chain variable domains (aVH) or VNAR moieties derived from sharks (see, for example, Hasler et al., Mol. Immunol. 75:28-37, 2016).

[0125] Techniques for producing single-domain antibodies (e.g., DAB or VHH) are known in the art, as disclosed, for example, in Cossins et al. (2006, Prot Express Purif 51:253-259) and Li et al. (Immunol. Lett. 188:89-95, 2017). Single-domain antibodies may be obtained, for example, from camels, alpacas, or llamas by standard immunization techniques. (See, for example, Muyldermans et al., TIBS 26:230-235, 2001; Yau et al., J Immunol Methods 281:161-75, 2003; and Maass et al., J Immunol Methods 324:13-25, 2007). VHH can possess potent antigen-binding ability and can interact with novel epitopes inaccessible to conventional VH-VL pairs (see, e.g., Muyldermans et al., 2001). Alpaca serum IgG contains approximately 50% heavy-chain-only IgG antibodies (HCAb) derived from camelids (see, e.g., Maass et al., 2007). Alpacas may be immunized with antigens, allowing for the isolation of VHH that bind to and neutralize target antigens (see, e.g., Maass et al., 2007). PCR primers for amplifying alpaca VHH coding sequences have been identified and may be used to construct alpaca VHH phage display libraries, which can then be used to isolate antibody fragments using standard biopanning techniques well known in the art (see, e.g., Maass et al., 2007).

[0126] In some embodiments, the PTK7 antibody or its antigen-binding moiety is part of a bispecific or multispecific binding agent. Bispecific and multispecific antibodies include: scFv1-ScFv2, ScFv12-Fc-scFv22, IgG-scFv, DVD-Ig, triomab / quadroma, two-in-one IgG, scFv2-Fc, TandAb, and scFv-HSA-scFv. In some embodiments, IgG-scFv is IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, svFc-(L)IgG, 2scFV-IgG, or IgG-2scFv. For example, see Brinkmann and Kontermann, MAbs 9(2):182-212 (2017); Wang et al., Antibodies, 2019, 8, 43; Dong et al., 2011, MAbs 3:273-88; Natsume et al., J. Biochem. 140(3):359-368, 2006; Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014; and Bates and Power, Antibodies, 2019, 8, 28.

[0127] Modification of VH and VL regions With respect to the VH and VL amino acid sequences, those skilled in the art will recognize that a "conservatively modified variant" is one in which a single amino acid or a small percentage of amino acids in the encoded sequence are altered by individual substitutions, deletions, or additions (insertions) to the nucleic acid or amino acids within the polypeptide encoding VH or VL, and that this alteration replaces an amino acid with a chemically similar amino acid (conservative amino acid substitution), and that the modified polypeptide retains its ability to specifically bind to PTK7.

[0128] In some embodiments, a conservatively modified variant of the PTK7 antibody or its antigen-binding moiety may have modifications in the framework region (i.e., outside the CDR), for example, the conservatively modified variant of the PTK7 antibody may have modifications in the amino acid sequences of the VH and VL CDRs (amino acid sequences (i) SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, FAS, and SEQ ID NO:7 respectively; (ii) SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, RTS, and SEQ ID NO:14 respectively; (iii) SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, KVS, and SEQ ID NO:22 respectively; (iv) SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, KVS, and SEQ ID The sequences have NO:27, and (v) shown in the set of SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, and SEQ ID NO:33 respectively, and have at least one conserved amino acid substitution in the framework region (FR). In some embodiments, the VH and VL amino acid sequences have a total of 8, 6, 4, 2, or 1 or fewer conserved amino acid substitutions in the FR compared to the amino acid sequences of the unmodified VH and VL regions. In some embodiments, the VH and VL amino acid sequences have 8-1, 6-1, 4-1, or 2-1 conserved amino acid substitutions in the FR compared to the amino acid sequences of the unmodified VH and VL regions. In further aspects of any of these embodiments, a conservedly modified variant of the PTK7 antibody, its antigen-binding moiety, or other binding substance exhibits specific binding to PTK7.

[0129] Conservative amino acid substitutions can be made by replacing a given amino acid with a residue having similar physicochemical properties, for example, by substituting one aliphatic residue with another (e.g., Ile, Val, Leu, or Ala with each other) or by substituting one polar residue with another (e.g., between Lys and Arg, Glu and Asp, or Gln and Asn). Other such conservative amino acid substitutions, such as substitutions of entire regions having similar hydrophobic properties, are well known. Polypeptides containing conservative amino acid substitutions can be tested with any of the assays described herein to confirm that the desired activity, e.g., the antigen-binding activity and specificity of the native or reference polypeptide, i.e., for PTK7, is retained.

[0130] In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance can be further optimized to reduce the potential for immunogenicity or to optimize other functional properties while maintaining functional activity, for example, for therapeutic use in humans. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance comprises a heavy-chain variable (VH) region and a light-chain variable (VL) region, the VH and VL regions having amino acid sequences represented by pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, SEQ ID NO:15 and SEQ ID NO:16, SEQ ID NO:34 and SEQ ID NO:35, SEQ ID NO:41 and SEQ ID NO:42, and SEQ ID NO:48 and SEQ ID NO:49, respectively, wherein the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance comprises a heavy-chain variable (VH) region and a light-chain variable (VL) region, the VH and VL regions having amino acid sequences represented by pairs of amino acid sequences selected from SEQ ID NO:1 and SEQ ID NO:2, SEQ ID NO:8 and SEQ ID NO:9, SEQ ID NO:15 and SEQ ID NO:16, SEQ ID NO:34 and SEQ ID NO:35, SEQ ID NO:41 and SEQ ID NO:42, and SEQ ID NO:48 and SEQ ID NO:49, respectively, wherein the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, wherein the CDR of the heavy-chain or light-chain variable region is not modified.

[0131] In some embodiments, a PTK7 antibody or its antigen-binding moiety or other binding material is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2, respectively, and the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, wherein the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, a PTK7 antibody or its antigen-binding moiety or other binding substance is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:1 and SEQ ID NO:2, respectively, and the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, wherein the CDR of the heavy-chain or light-chain variable region is not modified.

[0132] In some embodiments, a PTK7 antibody or its antigen-binding moiety or other binding material is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:8 and SEQ ID NO:9, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, a PTK7 antibody or its antigen-binding moiety or other binding substance is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:8 and SEQ ID NO:9, respectively, and the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, wherein the CDR of the heavy-chain or light-chain variable region is not modified.

[0133] In some embodiments, a PTK7 antibody or its antigen-binding moiety or other binding material is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:15 and SEQ ID NO:16, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, a PTK7 antibody or its antigen-binding moiety or other binding substance is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:15 and SEQ ID NO:16, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions within the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified.

[0134] In some embodiments, a PTK7 antibody or its antigen-binding moiety or other binding substance is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:34 and SEQ ID NO:35, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, a PTK7 antibody or its antigen-binding moiety or other binding substance is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:34 and SEQ ID NO:35, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions within the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified.

[0135] In some embodiments, a PTK7 antibody or its antigen-binding moiety or other conjugate is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:41 and SEQ ID NO:42, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, where the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, a conjugate is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:41 and SEQ ID NO:42, respectively, where the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, where the CDR of the heavy-chain or light-chain variable region is not modified.

[0136] In some embodiments, a PTK7 antibody or its antigen-binding moiety or other conjugate is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) VL region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:48 and SEQ ID NO:49, respectively, and the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 conservative amino acid substitutions in the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified. In some embodiments, a conjugate is provided herein, comprising a heavy-chain variable (VH) region and a light-chain variable (VL) region, wherein the VH and VL regions have amino acid sequences shown in SEQ ID NO:48 and SEQ ID NO:49, respectively, and the heavy-chain and light-chain variable framework regions may be modified by 1-8, 1-6, 1-4, or 1-2 amino acid substitutions, deletions, or insertions in the framework region, and the CDR of the heavy-chain or light-chain variable region is not modified.

[0137] In any of these embodiments, the functional activity of the PTK7-conjugated antibody or its antigen-binding moiety or other binding substance includes specific binding to PTK7. Additional functional activity includes depletion of PTK7+ cells (e.g., cancer cells or autoimmune cells). Where dose-dependency exists, it does not need to be identical to that of the reference antibody or its antigen-binding moiety, but rather needs to be substantially similar to or better than the dose-dependency in a given activity compared to the reference antibody or its antigen-binding moiety described herein (i.e., the candidate polypeptide will exhibit higher activity than the reference antibody).

[0138] Regarding conservative substitutions, amino acids can be grouped according to the similarity of their side chain properties (in AL Lehninger, in Biochemistry, second ed., pp. 73-75, Worth Publishers, New York (1975)): (1) Nonpolar: Ala(A), Val(V), Leu(L), Ile(I), Pro(P), Phe(F), Trp(W), Met(M); (2) Uncharged polar: Gly(G), Ser(S), Thr(T), Cys(C), Tyr(Y), Asn(N), Gln(Q); (3) Acidic: Asp(D), Glu(E); and (4) Basic: Lys(K), Arg(R), His(H).

[0139] Alternatively, for conserved substitutions, naturally occurring residues can be grouped based on common side-chain properties: (1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that affect chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe. Non-conservative substitutions would involve replacing members from one of these classes or another.

[0140] Specific conservative substitutions include, for example, Ala to Gly or Ser; Arg to Lys; Asn to Gln or His; Asp to Glu; Cys to Ser; Gln to Asn; Glu to Asp; Gly to Ala or Pro; His to Asn or Gln; Ile to Leu or Val; Leu to Ile or Val; Lys to Arg, Gln or Glu; Met to Leu, Tyr or Ile; Phe to Met, Leu or Tyr; Ser to Thr; Thr to Ser; Trp to Tyr; Tyr to Trp; and / or Phe to Val, Ile or Leu.

[0141] In some embodiments, a conservatively modified variant of the PTK7 antibody or its antigen-binding moiety is preferably at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a reference VH or VL sequence, where the CDR of the VH and VL sequences is not modified. The homology (percent identity) between the reference and modified sequences can be determined, for example, by comparing the two sequences using a freely available computer program commonly used for this purpose on the World Wide Web (e.g., BLASTp or BLASTn with default settings).

[0142] In some embodiments, the VH and VL amino acid sequences have a total of 8, 6, 4, 2, or 1 or fewer conserved amino acid substitutions in the framework region compared to the amino acid sequences of the unmodified VH and VL regions. In some embodiments, the VH and VL amino acid sequences have a total of 8-1, 6-1, 4-1, or 2-1 conserved amino acid substitutions in the framework region compared to the amino acid sequences of the unmodified VH and VL regions. In some embodiments, the VH and VL amino acid sequences have a total of 8, 6, 4, 2, or 1 or fewer amino acid substitutions, deletions, or insertions in the framework region compared to the amino acid sequences of the unmodified VH and VL regions. In some embodiments, the VH and VL amino acid sequences have a total of 8-1, 6-1, 4-1, or 2-1 conserved amino acid substitutions in the framework region compared to the amino acid sequences of the unmodified VH and VL regions. In some embodiments, the VH and VL amino acid sequences have a total of 8, 6, 4, 2, or 1 or fewer amino acid substitutions, deletions, or insertions compared to the amino acid sequences of the unmodified VH and VL regions.

[0143] Modification of a native (or reference) amino acid sequence can be achieved by any of the many methods known to those skilled in the art. Mutations can be introduced to a specific locus, for example, by synthesizing an oligonucleotide containing the desired mutant sequence and having restriction sites at both ends that allow ligation to a fragment of the native sequence. After ligation, the resulting reconstructed sequence encodes a mutant having the desired amino acid insertion, substitution, or deletion. Alternatively, oligonucleotide-directed site-specific mutagenesis can be employed to provide a modified nucleotide sequence in which specific codons are altered according to the desired substitution, deletion, or insertion. The techniques for making such modifications are very well established, including, for example, those disclosed by Walder et al. (Gene 42:133, 1986); Bauer et al. (Gene 37:73, 1985); Craik (BioTechniques, January 1985, 12-19); Smith et al. (Genetic Engineering: Principles and Methods, Plenum Press, 1981); and U.S. Patents No. 4,518,584 and No. 4,737,462, which are incorporated herein by reference in their entirety.

[0144] Steady-state region In some embodiments, the PTK7 antibody or its antigen-binding portion or another conjugate has a fully human constant region. In some embodiments, the PTK7 antibody or its antigen-binding portion or another conjugate has a humanized constant region. In some embodiments, the PTK7 antibody or its antigen-binding portion or another conjugate has a non-human constant region. The immunoglobulin constant region refers to the heavy chain or light chain constant region. The amino acid sequences of the human heavy chain and light chain constant regions are known in the art. The constant region may be any preferred type that can be selected from the classes of immunoglobulins such as IgA, IgD, IgE, IgG, and IgM. Some immunoglobulin classes can be further divided into isotypes, e.g., IgG1, IgG2, IgG3, IgG4, or IgAl and IgA2. The heavy chain constant regions (Fc) corresponding to different immunoglobulin classes are α, δ, ε, γ, and μ, respectively. The light chain may be either kappa (or κ) or lambda (or λ).

[0145] In some embodiments, the constant region may have an IgG1 isotype. In some embodiments, the constant region may have an IgG2 isotype. In some embodiments, the constant region may have an IgG3 isotype. In some embodiments, the constant region may have an IgG4 isotype. In some embodiments, the Fc domain may have a hybrid isotype comprising constant regions from two or more isotypes. In some embodiments, the immunoglobulin constant region may be an IgG1 or IgG4 constant region. In some embodiments, the PTK7 antibody heavy chain is of the IgG1 isotype and has the amino acid sequence shown in SEQ ID NO: 56. In some embodiments, the PTK7 antibody light chain is of the κ isotype and has the amino acid sequence shown in SEQ ID NO: 57.

[0146] Furthermore, the PTK7 antibody or its antigen-binding moiety or other binding material may be part of a larger binding material formed by covalent or non-covalent bonding between the antibody or antigen-binding moiety and one or more other proteins or peptides. Examples of such binding materials include the use of the streptavidin core region for preparing tetrameric scFv molecules (Kipriyanov, SM, et al. (1995), Human Antibodies and Hybridomas 6:93-101) and the use of cysteine ​​residues, marker peptides, and C-terminal polyhistidinyl peptides, such as the hexahistidinyl tag (disclosed as SEQ ID NO:59, "hexahistidinyl tag") for producing divalent biotinylated scFv molecules (Kipriyanov, SM, et al. (1994) Mol. Immunol. 31:10471058).

[0147] Fc domain modification to alter effect pedal functionality In some embodiments, the Fc region or Fc domain of the PTK7 antibody or its antigen-binding moiety or other binding substance does not substantially bind to at least one Fc receptor selected from FcyRI(CD64), FcyRIIA(CD32a), FcyRIIB(CD32b), FcyRIIIA(CD16a), and FcyRIIIB(CD16b). In some embodiments, the Fc region or domain does not substantially bind to any of the Fc receptors selected from FcyRI(CD64), FcyRIIA(CD32a), FcyRIIB(CD32b), FcyRIIIA(CD16a), and FcyRIIIB(CD16b). As used herein, “substantially unbound” means weak or absent binding to the selected Fc gamma receptor. In some embodiments, “substantially unbound” means at least a 1000-fold decrease in binding affinity to the Fc gamma receptor (i.e., an increase in Kd). In some embodiments, the Fc domain or region is Fc null. In this specification, “Fc null” means an Fc region or domain that has weak or no binding to any Fc gamma receptor. In some embodiments, the Fc null domain or region exhibits at least a 1000-fold decrease in binding affinity to the Fc gamma receptor (i.e., an increase in Kd).

[0148] In some embodiments, the Fc domain exhibits reduced or substantially absent effector functional activity. As used herein, “effector functional activity” refers to antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), and / or complement-dependent cell-mediated cytotoxicity (CDC). In some embodiments, the Fc domain exhibits reduced ADCC, ADCP, or CDC activity compared to a wild-type Fc domain. In some embodiments, the Fc domain exhibits reduced ADCC, ADCP, and CDC activity compared to a wild-type Fc domain. In some embodiments, the Fc domain exhibits substantially no effector function (i.e., the ability to stimulate or cause ADCC, ADCP, or CDC to act). As used herein, “substantially absent effector function” means that the effector functional activity is at least 1000-fold reduced compared to a wild-type or reference Fc domain.

[0149] In some embodiments, the Fc domain has reduced or no ADCC activity. As used herein, reduced or no ADCC activity means that the ADCC activity of the Fc domain is at least 10, at least 20, at least 30, at least 50, at least 100, or at least 500 times lower.

[0150] In some embodiments, the Fc domain has reduced or no CDC activity. As used herein, reduced or no CDC activity means that the CDC activity of the Fc domain is at least 10, at least 20, at least 30, at least 50, at least 100, or at least 500 times lower.

[0151] In vitro and / or in vivo cytotoxicity assays can be performed to confirm decreased / absent ADCC and / or CDC activity. For example, an Fc receptor (FcR) binding assay can be performed to confirm that the antibody does not bind to the Fcγ receptor (and therefore is likely to be absent ADCC activity). NK cells, the main cells that mediate ADCC, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays for evaluating the ADCC activity of target molecules are described in U.S. Patent No. 5,500,362 (see, e.g., Hellstrom, I. et al., Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I. et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985), and U.S. Patent No. 5,821,337 (see, Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, non-radioactive assay methods can be employed (see, for example, the ACTI® non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. Mountain View, Calif.) and the CytoTox 96® non-radioactive cytotoxicity assay (Promega, Madison, Wis.)). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively, the ADCC activity of the molecule of interest can be evaluated in vivo in animal models, such as those disclosed in, for example, Clynes et al., Proc. Nat'l Acad. Sci. USA 95:652-656 (1998).

[0152] A C1q binding assay may also be performed to confirm whether CDC activity is lost or reduced due to the inability of the antibody or Fc domain or region to bind to C1q. See, for example, the C1q and C3c binding ELISAs in International Publication No. 2006 / 029879 and International Publication No. 2005 / 100402. A CDC assay may also be performed to evaluate complement activation (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103:2738-2743 (2004)).

[0153] In some embodiments, the Fc domain has reduced or no ADCP activity. As used herein, reduced or no ADCP activity means that the ADCP activity of the Fc domain is at least 10, at least 20, at least 30, at least 50, at least 100, or at least 500 times lower.

[0154] ADCP binding assays may also be performed to confirm whether the antibody or Fc domain or region lacks ADCP activity or has low ADCP activity. See, for example, U.S. Patent Application Publication No. 20190079077 and U.S. Patent Application Publication No. 20190048078.

[0155] PTK7 antibodies or their antigen-binding moieties or other binding agents include those in which one or more Fc region residues, such as 238, 265, 269, 270, 297, 327, and 329, are substituted according to Kabat's EU numbering (see, for example, U.S. Patent No. 6,737,056). Such Fc variants include those with substitutions in two or more of the amino acid positions 265, 269, 270, 297, and 327 of the Kabat EU numbering, and the so-called "DANA" Fc variant in which residues 265 and 297 are substituted with alanine (see U.S. Patent No. 7,332,581). Certain antibody variants with reduced binding to FcR are also known. (See, for example, U.S. Patent No. 6,737,056, International Publication No. 2004 / 056312, and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001)). PTK7 antibodies or their antigen-binding moieties or other binding agents with reduced binding to FcR, including such amino acid modifications, can be prepared.

[0156] In some embodiments, the PTK7 antibody or its antigen-binding moiety or other binding substance includes an Fc domain or region having one or more amino acid substitutions that reduce FcγR binding, e.g., substitutions at positions 234 and 235 of the Fc region (residue EU numbering). In some embodiments, according to Kabat EU numbering, the substitutions are L234A and L235A (LALA). In some embodiments, the Fc domain includes D265A and / or P329G in the Fc region derived from the human IgG1 Fc region, according to Kabat EU numbering. In some embodiments, the substitutions are L234A, L235A and P329G (LALA-PG) in the Fc region derived from the human IgG1 Fc region, according to Kabat EU numbering. (See, for example, International Publication No. 2012 / 130831). In some embodiments, the substitutions are L234A, L235A, and D265A (LALA-DA) in the Fc region derived from the human IgG1 Fc region, according to Kabat's EU numbering.

[0157] In some embodiments, modifications are made to the Fc region that result in alterations (i.e., reductions) to C1q binding and / or complement-dependent cell-mediated cytotoxicity (CDC), as described in U.S. Patent No. 6,194,551, International Publication No. 99 / 51642, and Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

[0158] Methods for producing antibodies, antigen-binding sites, and other binding substances. In various embodiments, PTK7 antibodies, their antigen-binding moieties, and other conjugates can be produced in cell lines derived from humans, mice, or other animals. Recombinant DNA expression can be used to produce PTK7 antibodies, their antigen-binding moieties, and other conjugates. This enables the production of PTK7 antibodies and various PTK7 antigen-binding moieties and other conjugates (including fusion proteins) in selected host species. The production of PTK7 antibodies, their antigen-binding moieties, and other conjugates in bacteria, yeast, transgenic animals, and chicken eggs also offers alternatives to cell-based production systems. A major advantage of transgenic animals is the potential for high yields from renewable resources.

[0159] In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO: 1, 8, 15, 34, 41, or 48. In some embodiments, the nucleic acid encodes a PTK7 VL polypeptide having the amino acid sequence shown in SEQ ID NO: 2, 9, 16, 35, 42, or 49. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO: 1, 8, 15, 34, 41, or 48. In some embodiments, the nucleic acid encodes a PTK7 VL polypeptide having the amino acid sequence shown in SEQ ID NO: 2, 9, 16, 35, 42, or 49. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO: 1. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO: 15. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:34. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:41. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:48. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:2. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:9. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:16. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:35. In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:42.In some embodiments, the nucleic acid encodes a PTK7 VH polypeptide having the amino acid sequence shown in SEQ ID NO:49.

[0160] In some embodiments, the nucleic acid encodes VH and VL polypeptides having amino acid sequences shown in SEQ ID NO: 1 and 2. In some embodiments, the nucleic acid encodes VH and VL polypeptides having amino acid sequences shown in SEQ ID NO: 8 and 9. In some embodiments, the nucleic acid encodes VH and VL polypeptides having amino acid sequences shown in SEQ ID NO: 15 and 16. In some embodiments, the nucleic acid encodes VH and VL polypeptides having amino acid sequences shown in SEQ ID NO: 34 and 35. In some embodiments, the nucleic acid encodes VH and VL polypeptides having amino acid sequences shown in SEQ ID NO: 41 and 42. In some embodiments, the nucleic acid encodes VH and VL polypeptides having amino acid sequences shown in SEQ ID NO: 48 and 49.

[0161] As used herein, the terms “nucleic acid,” “nucleic acid sequence,” “polynucleotide sequence,” or “nucleotide” refer to polymer molecules incorporating units of ribonucleic acid, deoxyribonucleic acid, or analogs thereof. Nucleic acids may be single-stranded or double-stranded. Single-stranded nucleic acids may be single-stranded nucleic acids of denatured double-stranded DNA. In some embodiments, nucleic acids may be cDNA, for example, nucleic acids deleting introns.

[0162] In some embodiments, a single nucleic acid molecule is provided that encodes both the light and heavy chains of a PTK7 antibody or its antigen-binding moiety. In other embodiments, a reference to a nucleic acid encoding a PTK7 antibody or its antigen-binding moiety includes one nucleic acid encoding the light chain of the PTK7 antibody or its antigen-binding moiety and another nucleic acid encoding the heavy chain. Similarly, in some embodiments, a single vector provided may encode both the light and heavy chains of a PTK7 antibody or its antigen-binding moiety. Alternatively, one vector may encode the light chain of a PTK7 antibody or its antigen-binding moiety, and the other vector may encode the heavy chain.

[0163] PTK7 antibodies, nucleic acid molecules encoding amino acid sequences of their antigen-binding moieties or other binding substances can be prepared by various methods known in the art. These methods include, but are not limited to, the preparation of synthetic nucleotide sequences encoding PTK7 antibodies, antigen-binding moieties or other binding substances. Furthermore, nucleotide sequences encoding PTK7 antibodies or antigen-binding moieties or other binding substances can be prepared using oligonucleotide-mediated (or site-directed) mutagenesis, PCR-mediated mutagenesis, and cassette mutagenesis. At least the nucleic acid sequences encoding PTK7 antibodies, their antigen-binding moieties, binding substances, or polypeptides described herein can be recombined with vector DNA according to prior art, for example, blunt or adherent ends for ligation, restriction enzyme digestion to provide suitable ends, packing of suitable aggregated ends, alkaline phosphatase treatment to avoid undesirable conjugation, and ligation by a suitable ligase or other techniques known in the art. Techniques for such operations are disclosed, for example, in Maniatis et al., Molecular Cloning, Lab. Manual (Cold Spring Harbor Lab. Press, NY, 1982 and 1989), and Ausubel et al., Current Protocols in Molecular Biology (John Wiley & Sons), 1987–1993, and can be used to construct nucleic acid sequences and vectors encoding PTK7 antibodies or their antigen-binding moieties or their VH or VL polypeptides or other binding materials.

[0164] Nucleic acid molecules such as DNA contain nucleotide sequences that carry regulatory information for transcription and translation. A polypeptide is said to be "expressible" if such sequences are "functionally bound" to nucleotide sequences that encode polypeptides. Functional binding is a linkage where the regulatory DNA sequence and the DNA sequence to be expressed (e.g., a PTK7 antibody or its antigen-binding portion or other binding agent) are linked in such a way that a recoverable amount of the polypeptide or antigen-binding portion can be expressed. The exact properties of the regulatory regions required for gene expression can vary from organism to organism, as is well known in similar techniques. See, for example, Sambrook et al., 1989; Ausubel et al., 1987–1993.

[0165] Therefore, the expression of the PTK7 antibody or its antigen-binding moiety described herein can occur in either prokaryotic or eukaryotic cells. Suitable hosts include bacterial or eukaryotic hosts, including yeast, insect, fungal, avian, and mammalian cells, either in vivo or in situ, or host cells derived from mammals, insects, birds, or yeast. Mammalian cells or tissues may be from humans, primates, hamsters, rabbits, rodents, cattle, pigs, sheep, horses, goats, dogs, or cats, but any other mammalian cells may be used. Furthermore, for example, in vivo synthesis of ubiquitin-transmembrane polypeptide fusion proteins can be achieved by using a yeast ubiquitin hydrolase system. The fusion proteins thus produced can be processed in vivo or purified and processed in vitro, enabling the synthesis of the PTK7 antibody or its antigen-binding moiety or other binding substances described herein that have a specific amino-terminal sequence. In addition, problems associated with the retention of start codon-derived methionine residues in direct expression by yeast (or bacteria) can be avoided. (See, for example, Sabin et al., 7 Bio / Technol. 705 (1989); Miller et al., 7 Bio / Technol. 698 (1989)). Recombinant PTK7 antibodies or their antigen-binding moieties or other binding agents can be obtained using one of a series of yeast gene expression systems incorporating promoters and termination elements from actively expressed genes encoding glycolytic enzymes, which are produced in large quantities when yeast is grown in glucose-rich media. Known glycolytic genes can also provide highly efficient transcriptional regulatory signals. For example, promoter and terminator signals from phosphoglycerate kinase genes can be used.

[0166] The production of PTK7 antibodies or their antigen-binding moieties or other binding substances in insects can be achieved, for example, by infecting an insect host with a baculovirus engineered to express a polypeptide by a method known to those skilled in the art. See Ausubel et al., 1987–1993.

[0167] In some embodiments, the introduced nucleic acid sequence (encoding the PTK7 antibody or its antigen-binding portion or other binding substances or their polypeptides) is incorporated into a plasmid or viral vector capable of autonomous replication within the recipient host cell. For this purpose, any of the wide variety of vectors available are known and readily available to those skilled in the art. See, for example, Ausubel et al., 1987–1993. Important factors in selecting a particular plasmid or viral vector include the ease with which recipient cells containing the vector can be recognized and selected from recipient cells without the vector; the desired copy number of the vector in a particular host; and whether it is desirable for the vector to be "returned" between host cells of different species.

[0168] Exemplary prokaryotic vectors known in the art include plasmids that can replicate in Escherichia coli (E. coli). Other gene expression elements useful for expressing DNA encoding the PTK7 antibody or its antigen-binding moiety or other binding substances include, but are not limited to, (a) viral transcription promoters and their enhancer elements, such as the SV40 early promoter (Okayama et al., 3 Mol. Cell. Biol. 280 (1983)), Rous sarcoma virus LTR (Gorman et al., 79 PNAS 6777 (1982)), and Moloney mouse leukemia virus LTR (Grosschedl et al., 41 Cell 885 (1985)), (b) splice regions and polyadenylation sites, such as those derived from the SV40 late region (Okayarea et al., 1983), and (c) polyadenylation sites, such as those present in SV40 (Okayama et al., 1983). The DNA genes encoding immunoglobulins can be expressed using the following as expression elements, as described by Liu et al. and Weidle et al. (51 Gene 21 (1987)): the SV40 early promoter and its enhancer, the mouse immunoglobulin H chain promoter enhancer, the SV40 late region mRNA splicing, the rabbit S globin intercalated sequence, the immunoglobulin and rabbit S globin polyadenylation sites, and the SV40 polyadenylation element.

[0169] For nucleotide sequences encoding immunoglobulins, the transcription promoter may be, for example, human cytomegalovirus, and the promoter enhancer may be cytomegalovirus and mouse / human immunoglobulin.

[0170] In some embodiments, for the expression of a DNA coding region in rodent cells, the transcription promoter may be a viral LTR sequence, and the transcription promoter enhancer may be either or both a mouse immunoglobulin heavy chain enhancer and a viral LTR enhancer, as well as polyadenylation and transcription termination regions. In other embodiments, a DNA sequence encoding another protein is combined with the above expression elements to achieve protein expression in mammalian cells.

[0171] Each coding region or gene fusion is incorporated into or inserted into an expression vector. Then, recipient cells capable of expressing the PTK7 variable region or its antigen-binding moiety or other binding agent are transfused either alone with a nucleotide encoding the PTK7 antibody or its antibody polypeptide or antigen-binding moiety or other binding agent, or co-transfused with a polynucleotide or other binding agent encoding the VH and VL chain coding regions. The transfused recipient cells are cultured under conditions that allow expression of the incorporated coding region, and the expressed antibody chain or complete antibody or antigen-binding moiety or other binding agent is recovered from the culture.

[0172] In some embodiments, nucleic acids containing a coding region encoding the PTK7 antibody or its antigen-binding moiety or other binding substance are incorporated into separate expression vectors, which are then used for simultaneous translocation into recipient host cells. Each vector may contain one or more selection genes. For example, in some embodiments, two selection genes are used: a first selection gene designed for selection in bacterial systems and a second selection gene designed for selection in eukaryotic systems, where each vector has a set of coding regions. This strategy results in a vector that first directs the generation of a nucleotide sequence in a bacterial system, enabling amplification. The DNA vector thus produced and amplified in the bacterial host is then used for simultaneous translocation into eukaryotic cells, enabling the selection of simultaneous translocation cells having the desired translocation nucleic acid (e.g., containing the heavy and light chains of the PTK7 antibody). Non-limiting examples of selection genes for use in bacterial systems include genes conferring ampicillin resistance and genes conferring chloramphenicol resistance. Selected genes for use in eukaryotic transfusions include the xanthine guanine phosphoribosyltransferase gene (denoted as gpt) and the Tn5-derived phosphotransferase gene (denoted as neo). Alternatively, fusion nucleotide sequences encoding the VH and VL chains can be incorporated into the same expression vector.

[0173] For the translocation of the expression vector and the production of the PTK7 antibody or its antigen-binding moiety or other binding substance, the recipient cell line may be a Chinese hamster ovary cell line (e.g., DG44) or myeloma cells. Myeloma cells can synthesize, construct, and secrete immunoglobulins encoded by the translocated immunoglobulin gene and possess an immunoglobulin glycosylation mechanism. For example, in some embodiments, the recipient cells are recombinant Ig-producing myeloma cells SP2 / 0. SP2 / 0 cells produce only immunoglobulins encoded by the translocated gene. Myeloma cells can be grown in culture or in the peritoneal cavity of mice, where immunoglobulins secreted from ascites fluid can be obtained.

[0174] Expression vectors encoding PTK7 antibodies or their antigen-binding moieties or other binding substances can be introduced into suitable host cells by any of a variety of suitable means, including biochemical means such as transformation, translocation, protoplast fusion, calcium phosphate precipitation, and combination with polycations such as diethylaminoethyl (DEAE) dextran, as well as mechanical means such as electroporation, direct microinjection, and particle guns. As is well known to those skilled in the art, see Johnston et al., 240 Science 1538 (1988).

[0175] Yeast has several advantages over bacteria in the production of immunoglobulin heavy and light chains. Yeast performs post-translational peptide modifications, including glycosylation. Numerous recombinant DNA strategies exist that utilize strong promoter sequences and high-copy-number plasmids for the production of desired proteins in yeast. Yeast recognizes the leader sequence of cloned mammalian gene products and secretes polypeptides containing the leader sequence (i.e., pre-polypeptides). See, for example, Hitzman et al., 11th Intl. Conf. Yeast, Genetics & Molec. Biol. (Montpelier, France, 1982).

[0176] Yeast gene expression systems can conventionally be evaluated for the degree of production, secretion, and stability of antibodies, as well as constructed PTK7 antibodies and their antigen-binding moieties, and other binding substances. Various yeast gene expression systems can be used that incorporate promoters and termination elements of genes encoding glycolytic enzymes, which are produced in large quantities when yeast is cultured in glucose-rich medium. Known glycolytic genes can also provide highly efficient transcriptional regulatory signals. For example, promoter and terminator signals of the phosphoglycerate kinase (PGK) gene can be used. Another example is the translation elongation factor 1α promoter from Chinese hamster cells. Several approaches can be employed to evaluate the optimal expression plasmid for immunoglobulin expression in yeast. See II DNA Cloning 45, (Glover, ed., IRL Press, 1985) and, for example, U.S. Patent Application Publication 2006 / 0270045A1.

[0177] Bacterial strains can also be used as hosts for the production of antibody molecules or their antigen-binding moieties or other binding substances described herein. These include E. coli K12 strains such as E. coli W3110, Bacillus species, Enterobacteria such as Salmonella typhimurium or Serratia marcescens, and various Pseudomonas species. In relation to these bacterial host cells, plasmid vectors containing replicons and regulatory sequences derived from a host cell-compatible species are used. The vectors have replication sites as well as specific genes that enable phenotypic selection in transformed cells. Various approaches can be used to evaluate expression plasmids for the production of PTK7 antibodies or their antigen-binding moieties or other binding substances in bacteria (see Glover, 1985; Ausubel, 1987, 1993; Sambrook, 1989; Colligan, 1992–1996).

[0178] Host mammalian cells can be grown in vitro or in vivo. Mammalian cells provide post-translational modifications to immunoglobulin molecules, including removal of leader peptides, folding and construction of VH and VL chains, glycosylation of antibody molecules, and secretion of functional antibodies and / or their antigen-binding sites or other binding substances.

[0179] Mammalian cells that may be useful as hosts for antibody protein production include, in addition to the lymphoid cells described above, fibroblast-derived cells such as Vero cells or CHO-K1 cells. Exemplary eukaryotic cells that may be used to express immunoglobulin polypeptides include COS cells, including COS7 cells; 293 cells, including 293-6E cells; CHO cells, including CHO--S cells and DG44 cells; PERC6® cells (Crucell); and NSO cells, not limited to these. In some embodiments, specific eukaryotic host cells are selected based on their ability to perform desired post-translational modifications to the heavy and / or light chains. For example, in some embodiments, CHO cells produce polypeptides with a higher degree of sialylation than the same polypeptides produced by 293 cells.

[0180] In some embodiments, one or more PTK7 antibodies or their antigen-binding moieties or other binding substances can be produced in vivo in animals manipulated or transfected with one or more nucleic acid molecules encoding polypeptides, according to any preferred method.

[0181] In some embodiments, antibodies or their antigen-binding moieties are produced in a cell-free system. Non-limiting examples of cell-free systems are described, for example, in Sitaraman et al., Methods Mol. Biol. 498: 229-44 (2009); Spirin, Trends Biotechnol. 22: 538-45 (2004); and Endo et al., Biotechnol. Adv. 21: 695-713 (2003).

[0182] Many vector systems are available for the expression of VH and VL chains in mammalian cells (see Glover, 1985). Complete antibodies can be obtained according to various approaches. As discussed above, it is possible to achieve intracellular association and linkage of VH and VL chains, and optionally related constant regions, by co-expressing them in the same cell, thereby obtaining a complete tetrameric H2L2 antibody or its antigen-binding portion. Co-expression can occur by using the same or different plasmids in the same host. Cells expressing both chains can be directly selected by incorporating the nucleic acids encoding the VH and VL chains or their antigen-binding sites into the same plasmid and then introducing this plasmid into cells. Alternatively, one chain, for example, the plasmid encoding the VL chain, can be introduced into cells first, and then the resulting cell line can be introduced with the VH chain plasmid containing a second selection marker. Cell lines that produce antibodies and their antigen-binding moieties via either pathway can be transfected with plasmids encoding peptides, VH, VL, or additional copies of VH and VL chains in combination with additional selection markers, thereby generating cell lines with improved properties, such as increased production of the constructed PTK7 antibody or its antigen-binding moiety or other binding substances, or improved stability of the transfected cell line.

[0183] Furthermore, plants are emerging as a simple, safe, and economical alternative expression system to recombinant antibody production based on large-scale cultures of microbial or animal cells. PTK7-conjugated antibodies, or their antigen-binding sites or other conjugates, can be expressed in plant cell cultures or conventionally cultivated plants. Expression in plants may be systemic, limited to intracellular plastids, or limited to seeds (endosperm). See, for example, U.S. Patent Application Publication 2003 / 0167531; U.S. Patent No. 6,080,560; U.S. Patent No. 6,512,162; and International Publication No. 0129242. Several plant-derived antibodies have reached advanced stages of development, including clinical trials (see, for example, Biolex, NC).

[0184] In the case of a complete antibody, the variable regions (VH and VL regions) of the PTK7 antibody are typically bound to at least a portion of the constant region (Fc) or domain of human immunoglobulin. Human constant region DNA sequences can be isolated from various human cells, such as immortalized B cells (International Publication No. 87 / 02671), using well-known procedures. PTK7-conjugated antibodies may contain both light-chain and heavy-chain constant regions. The heavy-chain constant region may include the CH1, hinge, CH2, CH3, and optionally the CH4 region. In some embodiments, the CH2 domain may be deleted or omitted.

[0185] Techniques described for the production of single-chain antibodies (see, for example, U.S. Patent No. 4,946,778; Bird, Science 242:423-42 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); and Ward et al., Nature 334:544-54 (1989), which are incorporated herein by reference in their entirety) can be adapted to produce single-chain antibodies that specifically bind to PTK7. Single-chain antibodies are formed by linking the heavy and light chain variable regions of the Fv region via amino acid crosslinks, yielding single-chain polypeptides. Techniques for assembling functional Fv moieties in E. coli can also be used (see, for example, Skerra et al., Science 242:1038-1041 (1988), which are incorporated herein by reference in their entirety).

[0186] In some embodiments, the antigen-binding region or other binding material comprises one or more scFvs. An scFv can be, for example, a variable region fusion protein in which the variable regions of the antibody's heavy chain (VH) and light chain (VL) are linked by a short linker peptide consisting of 10 to about 25 amino acids. The linker typically contains a large amount of glycine for flexibility and serine or threonine for solubility, and can either connect the N-terminus of VH to the C-terminus of VL, or vice versa. This protein maintains the specificity of the original antibody despite the removal of the constant region and the introduction of the linker. For example, scFv antibodies are described in Houston, JS, Methods in Enzymol. 203 (1991) 46-96. Methods for constructing scFv molecules and designing suitable peptide linkers are described, for example, in U.S. Patent No. 4,704,692; U.S. Patent No. 4,946,778; Raag and Whitlow, FASEB 9:73-80 (1995); and Bird and Walker, TIBTECH, 9: 132-137 (1991). scFv-Fc is described by Sokolowska-Wedzina et al., Mol. Cancer Res. 15(8):1040-1050, 2017.

[0187] In some embodiments, the antigen-binding moiety or other binding material is a single-domain antibody, where the antibody moiety consists of a single monomeric variable antibody region. Single-domain antibodies can be derived from variable domains (e.g., nanobodies or VHH regions) of antibody heavy chains derived from camelids. Furthermore, single-domain antibodies may also be autonomous human heavy chain variable domains (aVH) or shark-derived VNAR regions (see, for example, Hasler et al., Mol. Immunol. 75:28-37, 2016).

[0188] Techniques for generating single-domain antibodies (DABs or VHHs) are known in the art, as disclosed, for example, in Cossins et al. (2006, Prot Express Purif 51:253-259) and Li et al. (Immunol. Lett. 188:89-95, 2017). Single-domain antibodies may be obtained, for example, from camels, alpacas, or llamas by standard immunization techniques. (See, for example, Muyldermans et al., TIBS 26:230-235, 2001; Yau et al., J Immunol Methods 281:161-75, 2003; and Maass et al., J Immunol Methods 324:13-25, 2007). VHH can possess potent antigen-binding ability and can interact with epitopes inaccessible to conventional VH-VL pairs (see, e.g., Muyldermans et al., 2001). Alpaca serum IgG contains approximately 50% heavy-chain-only IgG antibodies (HCAb) derived from camelids (see, e.g., Maass et al., 2007). Alpacas may be immunized with antigens, and VHH that binds to and neutralizes target antigens can be isolated (see, e.g., Maass et al., 2007). PCR primers that amplify alpaca VHH coding sequences have been identified and can be used to construct alpaca VHH phage display libraries, which can then be used to isolate antibody fragments using standard biopanning techniques well known in the art (see, e.g., Maass et al., 2007).

[0189] Techniques for producing multispecific antibodies include, but are not limited to, the recombinant co-expression of two immunoglobulin heavy-light chain pairs with different specificities (e.g., Milstein and Cuello, Nature 305: 537 (1983)), International Publication No. 93 / 08829, and Traunecker et al., EMBO J. 10: 3655 (1991)), and the "knob-in-hole" technique (e.g., U.S. Patent No. 5,731,168; see Carter (2001), J Immunol Methods 248, 7-15). Multispecific antibodies are produced by manipulating the electrostatic steering effect to create antibody Fc heterodimer molecules (see, e.g., International Publication No. 2009 / 089004A1); crosslinking two or more antibodies or their antigen-binding moieties (see, e.g., U.S. Patent No. 4,676,980 and Brennan et al., Science, 229: 81 (1985)); using leucine zippers to produce bispecific antibodies (see, e.g., Kostelny et al., J. Immunol., 148(5):1547-1553 (1992)); using "diabody" techniques to create bispecific antibody moieties (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-stranded Fv(scFv) dimers (see, e.g., Gruber et al., J. See Immunol., 152:5368 (1994); and it can also be prepared by preparing a trispecific antibody, for example, as described in Tutt et al. J. Immunol. 147: 60 (1991).

[0190] Modified antibodies having three or more functional antigen-binding sites, including "octopus antibodies," can also serve as binding agents (see, for example, U.S. Patent Application Publication 2006 / 0025576A1).

[0191] The binding substances (e.g., antibodies or antigen-binding moieties) as used herein also include “dual-acting FAb” or “DAF” containing antigen-binding sites that bind to two different antigens (see, for example, U.S. Patent Application Publication 2008 / 0069820 and Bostrom et al., 2009, Science 323:1610-14). “Crosumab” antibodies are also included herein (see, for example, International Publication Nos. 2009 / 080251, 2009 / 080252, 2009 / 080253, 2009 / 080254, and 2013 / 026833).

[0192] In some embodiments, the binding substance contains different antigen-binding sites fused to one or the other of two subunits of the Fc domain, so that the two subunits of the Fc domain can be contained in two non-identical polypeptide chains. Co-recombination expression of these polypeptides followed by dimerization results in several possible combinations of the two polypeptides. Therefore, to improve the yield and purity of bispecific molecules in recombinant production, it would be advantageous to introduce modifications to the Fc domain of the binding substance that promote the association of the desired polypeptide.

[0193] Generally, this method involves replacing one or more amino acid residues at the interface of two Fc domains with charged amino acid residues, which makes homodimer formation electrostatically undesirable but heterodimer formation electrostatically desirable.

[0194] In some embodiments, the binding agent is a “bispecific T cell engager” or BiTE (see, e.g., International Publication Nos. 2004 / 106381, 2005 / 061547, 2007 / 042261, and 2008 / 119567). This approach utilizes two antibody variable domains located on a single polypeptide. For example, a single polypeptide chain may contain two single-chain Fv(scFv) portions, each having a variable heavy chain (VH) and a variable light chain (VL) domain, separated by a polypeptide linker of sufficient length to allow intramolecular association between the two domains. This single polypeptide further contains a polypeptide spacer sequence between the two scFvs. Each scFv recognizes a different epitope, and these epitopes may be specific to different proteins, both of which are bound by the BiTE.

[0195] Since bispecific T cell engagers are single polypeptides, they can be expressed using any prokaryotic or eukaryotic cell expression system known in the art, such as the CHO cell line. However, specific purification techniques (see, e.g., EP1691833) may be required to separate monomeric bispecific T cell engagers from other multimeric species that may have biological activities other than the intended monomeric activity. In one exemplary purification scheme, a solution containing the secreted polypeptide is first subjected to metal affinity chromatography to elute the polypeptide using an imidazole concentration gradient. This eluate is further purified using anion exchange chromatography to elute the polypeptide using a sodium chloride concentration gradient. Finally, this eluate is subjected to size exclusion chromatography to separate the monomer from the multimeric species. In some embodiments, the conjugate is a bispecific antibody consisting of a single polypeptide chain containing two single-chain FV portions (scFVs) fused together by a peptide linker.

[0196] In some embodiments, the binding substance is multispecific, such as IgG-scFV. The IgG-scFv format includes IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, svFc-(L)IgG, 2scFV-IgG, and IgG-2scFv. These and other bispecific antibody formats and methods for producing them are described, for example, in Brinkmann and Kontermann, MAbs 9(2):182-212 (2017); Wang et al., Antibodies, 2019, 8, 43; Dong et al., 2011, MAbs 3:273-88; Natsume et al., J. Biochem. 140(3):359-368, 2006; Cheal et al., Mol. Cancer Ther. 13(7):1803-1812, 2014; and Bates and Power, Antibodies, 2019, 8, 28.

[0197] IgG-like bivariable domain antibody (DVD-Ig) is described in Wu et al., 2007, Nat Biotechnol 25:1290-97; Hasler et al., Mol. Immunol. 75:28-37, 2016, and International Publication Nos. 08 / 024188 and 07 / 024715. Triomab is described in Chelius et al., MAbs 2(3):309-319, 2010. 2-in-1-IgG is described in Kontermann et al., Drug Discovery Today 20(7):838-847, 2015. Tanden antibody or TandAb is described in the same publication by Kontermann et al. ScFv-HSA-scFv antibody is also described in the same publication by Kontermann et al.

[0198] Complete (e.g., whole) antibodies, their dimers, individual light and heavy chains, or their antigen-binding moieties and other conjugates can be recovered and purified by known techniques such as immunoadsorption or immunoaffinity chromatography, chromatographic methods such as HPLC (high-performance liquid chromatography), ammonium sulfate precipitation, gel electrophoresis, or any combination thereof. For an overview, see Scopes, Protein Purification (Springer-Verlag, NY, 1982). Substantially pure PTK7 antibodies or their antigen-binding moieties or other conjugates with at least approximately 90%–95% homogeneity are as advantageous as those with 98%–99% or higher homogeneity, especially for pharmaceutical applications. Once partially or to the desired level of homogeneity, complete PTK7 antibodies or their antigen-binding moieties or other conjugates can be used for therapeutic purposes or in the development and implementation of assay procedures, immunofluorescence staining, etc. For an overview, please refer to Vols. I & II Immunol. Meth. (Lefkovits & Pernis, eds., Acad. Press, NY, 1979 and 1981).

[0199] Antibody-drug conjugates In some embodiments, the PTK7 antibody, antigen-binding moiety, or other conjugate described herein is part of a PTK7 antibody-drug conjugate (also known as a PTK7 complex or PTK7 ADC). In some embodiments, the PTK7 antibody, antigen-binding moiety, or other conjugate is bound to at least one linker, and at least one drug is bound to each linker. As used herein, in the context of a complex, the term “drug” refers to cytotoxic substances (such as chemotherapeutic agents or drugs), immunomodulatory substances, nucleic acids (including siRNA), growth inhibitors, toxins (e.g., prototoxins, bacterial, fungal, plant or animal-derived enzyme-active toxins, or fragments thereof), radioisotopes, PROTACs, and other compounds that become active against target cells when delivered to target cells.

[0200] cytotoxic substances In some embodiments, the PTK7 complex comprises at least one drug (or “drug unit”) which is a cytotoxic substance. “Cytotoxic substance” refers to a drug that has a cytotoxic effect on cells. “Cytotoxic effect” refers to the depletion, elimination, and / or death of target cells. Cytotoxic substances include, for example, tubulin disruptors, topoisomerase inhibitors, DNA subgroove binders, and DNA alkylating agents.

[0201] Tubulin-destroying agents include, for example, auristatin, drastatin, tubulicin, colchicine, vinca alkaloids, taxanes, cryptophycin, maytansinoids, hemiasterin, and other tubulin-destroying agents. Auristatin is a derivative of the natural product drastatin 10. Examples of auristatin include MMAE (N-methylvaline-valine-dolaisoleuine-dolaproin-norephedrine), MMAF (N-methylvaline-valine-dolaisoleuine-dolaproin-phenylalanine), and AFP (see International Publication Nos. 2004 / 010957 and International Publication Nos. 2007 / 008603). Other auristatin-like compounds are disclosed, for example, in U.S. Patent Application Publication 2021 / 0008099, U.S. Patent Application Publication 2017 / 0121282, U.S. Patent Application Publication 2013 / 0309192, and U.S. Patent Application Publication 2013 / 0157960. Drastatin includes, for example, Drastatin 10 and Drastatin 15 (see, for example, Pettit et al., J. Am. Chem. Soc., 1987, 109, 6883-6885; Pettit et al., Anti-Cancer Drug Des., 1998, 13, 243-277; and U.S. Patent Application Publication 2001 / 0018422). Additional drastatin derivatives intended for use in the present invention are disclosed in U.S. Patent No. 9,345,785, which is incorporated herein by reference. In some embodiments, the tubulin disruptor is MMAE.

[0202] Tubricin non-limitingly includes tubulicin D, tubulicin M, tubulophenylalanine, and tubulotyrosine. International Publication No. 2017 / 096311 and International Publication No. 2016-040684 describe tubulicin analogs including tubulicin M.

[0203] Colchicine contains colchicine and CA-4 in an unrestricted manner.

[0204] Vinca alkaloids include, but are not limited to, vinblastine (VBL), vinorelbine (VRL), vincristine (VCR), and vindesine (VOS).

[0205] Taxanes include, but are not limited to, paclitaxel and docetaxel.

[0206] Cryptophycin contains cryptophycin-1 and cryptophycin-52 in an unrestricted manner.

[0207] Maytansinoids include, but are not limited to, maytansin; maytansinol; maytansin analogs in DM1, DM3, and DM4; and ansamatosin-2. Exemplary maytansinoid drug moieties include those having modified aromatic rings, e.g., C-19-dechloro(U.S. Patent No. 4,256,746) (prepared by reduction of ansamatosin P2 by lithium aluminum hydride); C-20-hydroxy(or C-20-demethyl)+ / -C-19-dechloro(U.S. Patents No. 4,361,650 and 4,307,016) (prepared by demethylation with streptomyces or actinomyces, or dechlorination with LAH); and C-20-demethoxy, C-20-acyloxy(--OCOR),+ / -dechloro(U.S. Patent No. 4,294,757) (prepared by acylation with acylchloride), and those having modifications at other positions.

[0208] The meitansinoid drug portion may have been modified, for example, C-9-SH (US Patent No. 4,424,219) (prepared by reaction of meitansinol with H2S or P2S5); C-14-alkoxymethyl (demethoxy / CH2OR) (US Patent No. 4,331,598); C-14-hydroxymethyl or acyloxymethyl (CH2OH or CH2OAc) (US Patent No. 4,450,254) (prepared from Nocardia); C-15-hydroxy / acyloxy (US Patent No. 4,364,866) (prepared by conversion of meitansinol by Streptomyces); C-15-methoxy (US Patent Nos. 4,313,946 and 4,315,929) (Trewia nudiflora). This also includes (isolated from nudiflora); C-18-N-demethyl (U.S. Patent Nos. 4,362,663 and 4,322,348) (prepared by demethylation of meitansinol by Streptomyces); and 4,5-deoxy (U.S. Patent No. 4,371,533) (prepared by titanium trichloride / LAH reduction of meitansinol).

[0209] Hemiasterine contains hemiasterine and HTl-286 in an unrestricted manner.

[0210] Other tubulin-destroying agents include takkaronolide A, takkaronolide B, takkaronolide AF, takkaronolide AJ, takkaronolide Al-epoxide, discodermold, epotilon A, epotilon B, and laurimalide.

[0211] In some embodiments, the cytotoxic substance may be a topoisomerase inhibitor such as camptothecin. Exemplary camptothecins include, for example, camptothecin, irinotecan (also known as CPT-11), berotecan, (7-(2-(N-isopropylamino)ethyl)camptothecin), topotecan, 10-hydroxy-CPT, SN-38, exatecan, and exatecan analog DXd (see U.S. Patent Publication No. 20150297748). Other camptothecins are disclosed in International Publication No. 1996 / 021666, International Publication No. 00 / 08033, U.S. Patent Publication No. 2016 / 0229862, and International Publication No. 2020 / 156189.

[0212] In some embodiments, the cytotoxic substance is duocarmcycin, including its synthetic analogs KW-2189 and CBI-TMI.

[0213] immunomodulator In some embodiments, the drug is an immunomodulator. The immunomodulator may be, for example, a TLR7 and / or TLR8 agonist, a STING agonist, a RIG-I agonist, or other immunomodulators.

[0214] In some embodiments, the drug is an immunomodulatory substance such as a TLR7 and / or TLR8 agonist. In some embodiments, the TLR7 agonist is selected from imidazoquinoline, imidazoquinolineamine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzoimidazole-2-amine, tetrahydropyridopyrimidine, heteroalothiadiazide-2,2-dioxide, benzonaphthyridine, guanosine analog, adenosine analog, thymidine homopolymer, ssRNA, CpG-A, polyG10, and polyG3. In some embodiments, the TLR7 agonist is selected from imidazoquinoline, imidazoquinolineamine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzoimidazole-2-amine, tetrahydropyridopyrimidine, heteroalothiadiazide-2,2-dioxide, or benzonaphthyridine. In some embodiments, the TLR7 agonist is a non-natural compound. Examples of TLR7 modifiers include GS-9620, GSK-2245035, imiquimod, reciquimod, DSR-6434, DSP-3025, IMO-4200, MCT-465, MEDI-9197, 3M-051, SB-9922, 3M-052, Limtop, TMX-30X, TMX-202, RG-7863, RG-7795, and U.S. Patent Publication No. 20160168164 (Janssen), U.S. Patent Publication No. 20150299194 (Roche), U.S. Patent Publication No. 20110098248 (Gilead Sciences), and U.S. Patent Publication No. 20100143301 (Gilead Sciences). This includes compounds disclosed in U.S. Patent Application Publication No. 20090047249 (Gilead Sciences).

[0215] In some embodiments, the TLR8 agonist is selected from benzoazepines, imidazoquinolines, thiazoloquinolines, aminoquinolines, aminoquinazolines, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzoimidazole-2-amine, tetrahydropyridopyrimidine, or ssRNA. In some embodiments, the TLR8 agonist is selected from benzoazepines, imidazoquinolines, thiazoloquinolines, aminoquinolines, aminoquinazolines, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzoimidazole-2-amine, and tetrahydropyridopyrimidine. In some embodiments, the TLR8 agonist is a non-natural compound. Examples of TLR8 agonists include motorimodo, resikimod, 3M-051, 3M-052, MCT-465, IMO-4200, VTX-763, and VTX-1463.

[0216] In some embodiments, the TLR8 agonist may be any of the compounds described in International Publication No. 2018 / 170179, International Publication No. 2020 / 056198, and International Publication No. 2020056194.

[0217] Other TLR7 and TLR8 agonists include, for example, International Publication No. 2016142250, International Publication No. 2017046112, International Publication No. 2007024612, International Publication No. 2011022508, International Publication No. 2011022509, International Publication No. 2012045090, International Publication No. 2012097173, International Publication No. 2012097177, International Publication Publication No. 2017079283, U.S. Patent Application Publication No. 20160008374, U.S. Patent Application Publication No. 20160194350, U.S. Patent Application Publication No. 20160289229, U.S. Patent No. 6043238, U.S. Patent Application Publication No. 20180086755 (Gilead), International Publication No. 2017216054 (Roche), International Publication No. 2017190669 (Shanghai) De Novo Pharmatech), International Publication No. 2017202704 (Roche), International Publication No. 2017202703 (Roche), International Publication No. 20170071944 (Gilead), U.S. Patent Application Publication No. 20140045849 (Janssen), U.S. Patent Application Publication No. 20140073642 (Janssen), International Publication No. 2014056953 (Janssen), International Publication No. 2014076221 (Janssen), International Publication No. 2014128189 (Janssen), U.S. Patent Application Publication No. 20140350031 (Janssen), International Publication No. 2014023813 (Janssen), U.S. Patent Application Publication No. 20080234251 (Array Array Biopharma, U.S. Patent Application Publication No. 20080306050 (Array Biopharma), U.S. Patent Application Publication No. 20100029585 (Ventirx Pharma), U.S. Patent Application Publication No. 20110092485 (Ventirx Pharma), U.S. Patent Application Publication No. 20110118235 (Ventirx Pharma), U.S. Patent Application Publication No. 20120082658 (Ventirx Pharma), U.S. Patent Application Publication No. 20120219615 (Ventirx Pharma), U.S. Patent Application Publication No. 20140066432 (Ventirx Pharma), U.S. Patent Application Publication No. 20140088085 (VentirxIt is disclosed in U.S. Patent Publication No. 20140275167 (Novira Therapeutics), U.S. Patent Publication No. 20130251673 (Novira Therapeutics), International Publication No. 2018198091 (Novartis AG), and U.S. Patent Publication No. 20170131421 (Novartis AG).

[0218] In some embodiments, immunomodulators are STING agonists. Examples of STING agonists include, for example, those disclosed in International Publication Nos. 2020059895, 2015077354, 2020227159, 2020075790, 2018200812, and 2020074004.

[0219] In some embodiments, immunomodulators are RIG-I agonists. Examples of RIG-I agonists include KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400, and KIN2000.

[0220] toxin In some embodiments, the drug is an enzyme-active toxin or a fragment thereof, non-limitingly comprising diphtheria A chain, unbound active fragment of diphtheria toxin, exotoxin A chain (derived from Pseudomonas aeruginosa), lysine A chain, abrin A chain, modexin A chain, alpha-sarcin, Aleurites fordii protein, dianthin protein, Phytolaca americana protein (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, geronin, mitogenin, restrictosin, phenomycin, enomycin, and trichothecenes.

[0221] radioactive isotope In some embodiments, the drug is a radioactive atom. Various radioactive isotopes are available for the production of radioactive complexes. Examples include I131, I125, Y90, Re186, Re188, Sm153, Bi213, P32, Pb212, and radioactive isotopes of lutetium (e.g., Lu177).

[0222] PROTAC In some embodiments, the drug is a proteolytic chimera (PROTAC). PROTACs are described, for example, in U.S. Patent Publication Nos. 20210015942, 20210015929, 20200392131, 20200216507, 20200199247, and 20190175612, the disclosures of which are incorporated herein by reference.

[0223] Linker A PTK7 complex typically contains at least one linker, to which at least one drug is bound. Typically, the complex contains a linker between the PTK7 antibody (or its antigen-binding portion or other binding substance) and the drug (sometimes called a "drug unit"). In various embodiments, the linker may be a protease-cleaving linker, an acid-cleaving linker, a disulfide linker, a disulfide-containing linker, or a disulfide-containing linker having a dimethyl group adjacent to the disulfide bond (e.g., SPDB linker) (see, e.g., Jain et al., Pharm. Res. 32:3526-3540 (2015); Chari et al., Cancer Res. 52:127-131 (1992); U.S. Patent No. 5,208,020), or a self-stabilizing linker (see, e.g., International Publication No. 2018 / 031690 and International Publication No. 2015 / 095755 and Jain et al., Pharm. Res. 32:3526-3540). The linkers may be non-cuttable linkers (see, for example, International Publication No. 2007 / 008603), photosensitive linkers, and / or hydrophilic linkers (see, for example, International Publication No. 2015 / 123679).

[0224] In some embodiments, the linker is a cleavable linker that can be cleaved under intracellular conditions, and cleavage of the linker releases the drug from the antibody (or its antigen-binding portion or other binding substance) and / or the linker in the intracellular environment. For example, in some embodiments, the linker can be cleaved by a cleaving agent present in the intracellular environment (e.g., within lysosomes, endosomes, or caveolae). The linker may be a peptidyl linker that can be cleaved by an intracellular peptidase or protease enzyme, for example, non-limitingly containing lysosome or endosomal proteases (see, for example, International Publication No. 2004 / 010957, U.S. Patent Application Publication No. 20150297748, U.S. Patent Application Publication No. 2008 / 0166363, U.S. Patent Application Publication No. 20120328564, and U.S. Patent Application Publication No. 20200347075). Typically, peptidyl linkers are at least one amino acid long or at least two amino acids long. Intracellular cleavage agents may include cathepsins B and D, as well as plasmin, all of which are known to hydrolyze dipeptide drug derivatives, resulting in the release of active drugs within target cells (see, e.g., Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123). Most typically, these are peptidyl linkers that can be cleaved by enzymes present in target antigen-expressing cells. For example, peptidyl linkers that can be cleaved by cathepsin B, a thiol-dependent protease highly expressed in cancer tissue (e.g., Phe-Leu or Gly-Phe-Leu-Gly linkers) can be used. Other such linkers are described, for example, in U.S. Patent No. 6,214,345. In certain embodiments, the peptidyl linker cleavable by intracellular proteases is a Val-Cit linker, a Phe-Lys linker (see, for example, U.S. Patent No. 6,214,345, which describes the synthesis of doxorubicin by a val-cit linker), or a Gly-Gly-Phe-Gly (SEQ ID NO: 60) linker (see, for example, U.S. Patent Application Publication No. 2015 / 0297748).One advantage of using intracellular proteolytic release of drugs is that, upon complexation, the drug is typically attenuated, and the serum stability of the complex is typically increased. See also U.S. Patent No. 9,345,785.

[0225] As used herein, the terms “intracellularly cleaved” and “intracellular cleavage” refer to intracellular metabolic processes or reactions to an antibody-drug conjugate in which the covalent bond between the drug (e.g., a cytotoxic substance) and the antibody, such as the linker, is cleaved, resulting in the release of the drug or other metabolites of the conjugate from the antibody within the cell. Thus, the cleaved portion of the conjugate is an intracellular metabolite.

[0226] In some embodiments, cleavable linkers are pH-sensitive, i.e., susceptible to hydrolysis at specific pH values. Typically, pH-sensitive linkers are hydrolyzable under acidic conditions. For example, acid-unstable linkers that are hydrolyzable within lysosomes (e.g., hydrazones, semicarbazones, thiosemicarbazones, cis-aconitamides, orthoesters, acetals, ketals, etc.) can be used. (See, for example, U.S. Patents 5,122,368; 5,824,805; and 5,622,929; Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123; Neville et al., 1989, Biol. Chem. 264:14653-14661). Such linkers are relatively stable under neutral pH conditions, such as in blood, but become unstable below pH 5.5 or 5.0, which is the approximate pH of lysosomes. In certain embodiments, the hydrolyzable linker is a thioether linker (for example, a thioether linked to a drug unit via an acylhydrazone bond (see, for example, U.S. Patent No. 5,622,929)).

[0227] In some embodiments, the linker is cleavable under reducing conditions (e.g., a disulfide linker). Various disulfide linkers are known, including, for example, SATA (N-succinimidyl-5-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), and SMPT (N-succinimidyloxycarbonyl-alpha-methyl-alpha-(2-pyridyldithio)toluene), and those that can be formed using SPDB and SMPT (see, for example, Thorpe et al., 1987, Cancer Res. 47:5924-5931; Wawrzynczak et al., In lmmunoconjugates: Antibody Conjugates in Radioimagery and Therapy of Cancer (CW Vogel ed., Oxford U. Press, 1987). See also U.S. Patent No. 4,880,935).

[0228] In some embodiments, the linker is a malonate linker (Johnson et al., 1995, Anticancer Res. 15:1387-93), a maleimidobenzoyl linker (Lau et al., 1995, Bioorg-Med-Chem. 3(10):1299-1304), or a 3'-N-amide analog (Lau et al., 1995, Bioorg-Med-Chem. 3(10):1305-12). In some embodiments, the linker is inclementable, such as a maleimidocaproyl linker, and the drug is released by antibody degradation. (See, for example, U.S. Patent Application Publication No. 2005 / 0238649).

[0229] In some embodiments, the linker is substantially insensitive to the extracellular environment. As used herein, “substantially insensitive to the extracellular environment” in the context of the linker means that, when the antibody-drug conjugate (ADC) is present in the extracellular environment (e.g., plasma), about 20% or less, typically about 15% or less, more typically about 10% or less, even more typically about 5% or less, about 3% or less, or about 1% or less of the linker in the ADC sample. Whether the linker is substantially insensitive to the extracellular environment can be determined, for example, by culturing both (a) an ADC ("ADC sample") and (b) equimolar amounts of an uncomplexed antibody or drug ("control sample") independently with plasma for a predetermined time (e.g., 2, 4, 8, 16, or 24 hours), and then comparing the amount of uncomplexed antibody or drug present in the ADC sample with that present in the control sample, as measured, for example, by high-performance liquid chromatography.

[0230] In some embodiments, the linker facilitates intracellular translocation. In some embodiments, the linker facilitates intracellular translocation when conjugated with a drug such as a cytotoxic substance (i.e., in the environment of the linker-drug portion of the ADC described herein). In yet another embodiment, the linker facilitates intracellular translocation when conjugated with both a drug and a PTK7 antibody (i.e., in the environment of the ADC described herein).

[0231] Various linkers that can be used in this composition and method are described, for example, in International Publication No. 2004010957. In some embodiments, the protease-cleaving linker comprises a thiol-reactive spacer and a dipeptide. In some embodiments, the protease-cleaving linker consists of a thiol-reactive maleimidocaproyl spacer, a valine-citrulline dipeptide, and a p-amino-benzyloxycarbonyl spacer.

[0232] In some embodiments, the acid-cleaving linker is a hydrazine linker or a quaternary ammonium linker (see, for example, International Publication No. 2017 / 096311 and International Publication No. 2016 / 040684).

[0233] In some embodiments, the linker is a self-stabilizing linker containing a maleimide group as described in U.S. Patent No. 9,504,756.

[0234] In some embodiments, the linker is a hydrophilic linker such as the hydrophilic peptide disclosed in International Publication No. 2015 / 123679, and the sugar alcohol polymer-based linker disclosed in International Publication No. 2013 / 012961 and International Publication No. 2019 / 213046.

[0235] In another embodiment, a conjugate of a PTK7 antibody (or antigen-binding moiety or other binding substance) and a drug can be prepared using a variety of bifunctional protein coupling agents such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), iminothiolane (IT), difunctional derivatives of imide esters (e.g., dimethylHCl adipimidoate), active esters (e.g., disuccinimidyl suberate), aldehydes (e.g., glutaraldehyde), bisazide compounds (e.g., bis(p-azidobenzoyl)hexanediamine), bisdiazonium derivatives (e.g., bis(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (e.g., toluene 2,6-diisocyanate), and bis-active fluorine compounds (e.g., 1,5-difluoro-2,4-dinitrobenzene). Chelating agents for the complexation of radioactive nucleotides with antibodies, their antigen-binding moieties, or other binding substances are described, for example, in International Publication No. 94 / 11026.

[0236] The PTK7 antibody (or antigen-binding moiety or other binding substance) conjugates include, but are not limited to, such conjugates prepared using crosslinking reagents that are commercially available (e.g., from Pierce Biotechnology, Inc., Rockford, IL., USA) and include, but are not limited to, BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, and sulfo-SMPB, and SVSB (succinimidyl-(4-vinylsulfone)benzoate).

[0237] In some embodiments, the linker may be bound to the end of the amino acid sequence of the antibody, antigen-binding moiety, or other binding substance, or to the side chain modification of the antibody, antigen-binding moiety, or other binding substance, such as lysine, serine, threonine, cysteine, tyrosine, aspartic acid, unnatural amino acid residues, glutamine, or glutamic acid residues. The binding between the antibody, antigen-binding moiety, or other binding substance and the linker or drug may be via one of a number of bonds, for example, non-limitingly, such as amide bonds, ester bonds, ether bonds, carbon-nitrogen bonds, carbon-carbon single, double, or triple bonds, disulfide bonds, or thioether bonds. Functional groups capable of forming such bonds include, for example, amino groups, carboxyl groups, aldehyde groups, azide groups, alkynes and alkene groups, ketones, carbonates, and carbonyl functional groups bound to leaving groups such as cyano, succinimidyl, and hydroxyl groups.

[0238] In some embodiments, the linker is bound to an antibody, antigen-binding moiety, or other binding substance at an interchain disulfide. In some embodiments, the linker is bound to an antibody, antigen-binding moiety, or other binding substance at a hinged cysteine ​​residue. In some embodiments, the linker is bound to an antibody, antigen-binding moiety, or other binding substance at an engineered cysteine ​​residue. In some embodiments, the linker is bound to an antibody, antigen-binding moiety, or other binding substance at a lysine residue. In some embodiments, the linker is bound to an antibody, antigen-binding moiety, or other binding substance at an engineered glutamine residue. In some embodiments, the linker is bound to an antibody, antigen-binding moiety, or other binding substance at an engineered non-native amino acid in the heavy chain.

[0239] In some embodiments, the linker is bound to the antibody, antigen-binding moiety, or other binding substance via a sulfhydryl group. In some embodiments, the linker is bound to the antibody, antigen-binding moiety, or other binding substance via a primary amine. In some embodiments, the linker is bound via a link formed by reacting a non-natural amino acid on the antibody, antigen-binding moiety, or other binding substance with an oxime bond formed by modifying a ketone group with an alkoxyamine on the drug.

[0240] In some embodiments, the linker is bound to an antibody, antigen-binding moiety, or other binding substance via a saltase A linker. The saltase A linker can be produced by the saltase A enzyme fusing the LPXTG recognition motif (SEQ ID NO: 61) to the N-terminal GGG motif to regenerate a native amide bond.

[0241] In some embodiments, the complex of the present disclosure comprises a PTK7 antibody, its antigen-binding moiety, and one of other binding substances, at least one linker bound to the binding substance, and at least one drug unit, wherein each drug unit is bound to the linker, the linker may comprise at least one polar group.

[0242] In some embodiments, in the complex of the present disclosure, the linker is derived from a linker compound or a stereoisomer or salt thereof, the linker compound comprising a linker unit, a stretcher group attached to the linker unit, an optional amino acid unit, and at least one polar group, wherein the stretcher group has a binding site to a binding substance and a binding site to an amino acid unit (if present) or a linker subunit, the amino acid unit (if present) has a binding site to a stretcher unit and a binding site to a linker unit, and the linker unit has a binding site to an amino acid unit (if present) or to a stretcher unit and a binding site to at least one drug unit.

[0243] Some of the components and variations of linkers (and linker compounds) are illustrated and demonstrated by the “Module of Linkers” provided herein.

[0244] Linker's characteristics The linkers (and linker compounds) of this disclosure are further illustrated by the following embodiments, which should not be construed as limiting. Appearance 1 .below: (a) Having 1 to 4 binding sites for drug units, and having the following structure (i) or (ii): A linker unit having one of TIFF2026534278000006.tif38128, (b) at least one polar group comprising polymer units, optionally sugar units, optionally carboxyl units, and combinations thereof, (c) Optionally, stretcher group having a binding site for PTK7 binding material A linker compound or its stereoisomer or salt, which includes During the ceremony, α- is a binding site to a group that can be cleaved by enzymes. β- is a binding site to at least one polar group, δ- is a binding site to H, at least one drug unit, or a binding site to a linking group attached to at least one drug unit. The polymer units include polyamides, polyethers, or combinations thereof, where the polyethers include hydroxyl groups, polyhydroxyl groups, sugar groups, carboxyl groups, or combinations thereof. Each R a These are independently H or C1-C6 alkyl groups. Each R b They are independent of Haro, C 1~6 An alkyl group, a binding site to at least one drug unit, or a binding site to at least one polar group, x is 0, 1, 2, 3, or 4. y is 0, 1, 2, or 3. R c The bonds are -C(O)-, -S(O)-, -SO2-, C 1~6 Alkilen, C 1~6 Alkinylene, triazolyl, or a combination thereof. Y is a bond, -O-, -S-, -N(R a -C(O)-, -S(O)-, -SO2-C1~C6 alkylene, C1~C6 alkenylene, C1~C6 alkynylene, triazolyl or combination thereof. Linker compounds or their stereoisomers or salts. Appearance 2 The linker unit has the following structure (ia) or (ii-a): The linker compound according to embodiment 1, having one of TIFF2026534278000007.tif38128 or a stereoisomer or salt thereof. Appearance 3 The linker unit has one of the following structures: (ib), (ic), (id), (ie), or (if): A linker compound according to embodiment 1 or 2, having one of TIFF2026534278000008.tif132128 or a stereoisomer or salt thereof. Pattern 4 The polar group is represented by the following formula: L3-N(CH2-(CH(XR)) k -X1(X2))2(X) or comprising at least one sugar unit having a stereoisomer or salt thereof, During the ceremony, Each X is independently selected from NH and O. Each R is independently selected from hydrogen, acetyl, monosaccharides, disaccharides, and polysaccharides. Each X1 is independently selected from CH2 and C(O), Each X2 is independently selected from H, OH, and OR. k is between 1 and 10. L3 is the bonding site to the rest of the polar group. A linker compound according to any one of embodiments 1 to 3. Appearance 5 At least one sugar unit has the following structure (XII) or (XIII): Having one of TIFF2026534278000009.tif70128 or its stereoisomer or salt, During the ceremony, Each R is independently selected from hydrogen, monosaccharides, disaccharides, and polysaccharides. m is 1 to 8. n is between 0 and 4. The linker compound according to embodiment 4. Appearance 6 .below: (a) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 (XX) or its stereoisomer, salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene. R 24 and R 25Each of these independently represents H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and an optional C3-C3 group. 10 Carbon rings, unsubstituted C1-C3 alkylenes, C3-C 10 A carbocyclic ring, an optionally substituted heteroaryl ring, an optionally substituted carbocyclic ring, a substituted-C1~C8 alkyl group, a substituted-C(O)-C1~C8 alkyl group, a chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 Select or -NR 24 R 25 They come together to form a C3-C8 heterocycle, n20 is 2-26; or (b) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 (XXI) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene. R 24 and R 25 One of them is H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, or possibly substituted C3-C 10 Carbon ring, unsubstituted C1-C3 alkylene C3~C 10 A carbocyclic ring, an optionally substituted heteroaryl ring, an optionally substituted carbocyclic ring, a substituted-C1~C8 alkyl group, a substituted-C(O)-C1~C8 alkyl group, a chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More selected, R 24 and R25 The other is polyethylene glycol which may have 1 to 24 ethylene glycol subunits. n20 is 2-26; or (c) ~R 20 -[-R 26 -[R 29 -[O-CH2-CH2-] n20 R 29 ] n21 -R 27 -NR 24 R 25 ] n27 (XXII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to or to a group that can be enzymatically cleaved; R 26 and R 27 Each is arbitrary, and the combination, C1~C 12 Alkylene, -NH-C1~C 12 Alkylene, -C1~C 12 Alkylene-NH-,-C1~C 12 Alkylene -N(CH3)-, -C(O)-C1~C 12 Alkylene, -C1~C 12 Alkylene-C(O)-,-NH-C1~C 12 Alkylene-C(O)- and -C(O)-C1~C 12 Selected independently from alkylene-NH-; R 24 and R 25 One of them is H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, or possibly substituted C3-C 10 Carbon ring, unsubstituted C1-C3 alkylene C3~C 10 A carbocyclic ring, an optionally substituted heteroaryl ring, an optionally substituted carbocyclic ring, a substituted-C1~C8 alkyl group, a substituted-C(O)-C1~C8 alkyl group, a chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28More selected, R 24 and R 25 The other side may be H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, or an optional C3-C 10 Carbon rings, unsubstituted C1-C3 alkylenes, C3-C 10 A carbocyclic ring, an optionally substituted heteroaryl ring, an optionally substituted carbocyclic ring, a substituted-C1~C8 alkyl group, a substituted-C(O)-C1~C8 alkyl group, a chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 , and selected from polyethylene glycol which may have 1 to 24 ethylene glycol subunits; or -NR 24 R 25 They come together to form a C3-C8 heterocycle, Each R 29 The elements are optional and independently selected from -C(O)-, -NH-, -C(O)-C1~C6alkylene-, -NH-C1~C6alkylene-, -C1~C6alkylene-NH-, -C1~C6alkylene-C(O)-, -NH(CO)-C1~C6alkylene-, -N(CH3)-(CO)-C1~C6alkylene-, -NH(CO)NH-, and triazole. n20 is between 2 and 26. n21 is 1 to 4. n27 is 1-4; or (d) ~R 20 -R 21 -[-C(R α )HC(O)-N(R N )-] n20 -R 22 -NR 24 R 25 (XXIII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21The bond is C1-C3 alkylene, -C1-C3 alkylene-[O-CH2-CH2-] n20 ,-[CH2-CH2-O] n20 -C1~C3 alkylene- or -C1~C3 alkylene-[O-CH2-CH2-] n20 -C(O)-, R 22 C1~C3 alkylene, -C1~C3 alkylene-[O-CH2-CH2-] n20 ,-[CH2-CH2-O] n20 -C1~C3 alkylene- or -C1~C3 alkylene-[O-CH2-CH2-] n20 -C(O)-, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N These are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 And, R 24 and R 25 Each of these independently represents H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and an optional C3-C3 group. 10 Carbon rings, unsubstituted C1-C3 alkylenes, C3-C 10 A carbocyclic ring, an optionally substituted heteroaryl ring, an optionally substituted carbocyclic ring, a substituted-C1~C8 alkyl group, a substituted-C(O)-C1~C8 alkyl group, a chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 Select or -NR 24 R 25 They come together to form a C3-C8 heterocycle, Each n20 is independently 2 to 26; or (e) ~R 20 -R 21 -[-C(R α )HC(O)-N(R N )-]n20 -R 22 -CO2R 26 (XXIV) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently a bond, a C1-C3 alkylene, or a -C1-C3 alkylene [O-CH2-CH2-] n20 And, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N These are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 And, R 24 and R 25 Each of these independently represents H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and an optional C3-C3 group. 10 Carbon rings, unsubstituted C1-C3 alkylenes, C3-C 10 A carbocyclic ring, an optionally substituted heteroaryl ring, an optionally substituted carbocyclic ring, a substituted-C1~C8 alkyl group, a substituted-C(O)-C1~C8 alkyl group, a chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 Select or -NR 24 R 25 They come together to form a C3-C8 heterocycle, R 26 is H or C1-C4 alkyl, Each n20 is independently between 2 and 26, However, at least one R α or R N ha-R 22 -NR 24 R25 is; or (f) ~R 20 -R 21 -[C(R α )HC(O)-N(R N )-] n20 -R 22 -N-(R 23 -NR 24 R 25 )2(XXV) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently a bond, a C1-C3 alkylene, or a -C1-C3 alkylene-[O-CH2-CH2-] n20 And, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N These are independently H or C1-C6 alkyl groups. Each R 23 These are independently C1-C6 alkylenes, R 24 and R 25 Each of these independently represents H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and an optional C3-C3 group. 10 Carbon rings, unsubstituted C1-C3 alkylenes, C3-C 10 A carbocyclic ring, an optionally substituted heteroaryl ring, an optionally substituted carbocyclic ring, a substituted-C1~C8 alkyl group, a substituted-C(O)-C1~C8 alkyl group, a chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 Select or -NR 24 R 25 They come together to form a C3-C8 heterocycle, Each n20 is independently between 2 and 26; A linker compound according to any one of embodiments 1 to 5, comprising a polar group having a more selected formula. Appearance 7 .R 24 and R 25 Each is independently selected from H and polyhydroxyl groups, however R 24 and R 25 The linker compound according to embodiment 6, wherein neither of the elements can become H. Appearance 8 The linker compound according to embodiment 6 or 7, wherein the polyhydroxyl group is a linear monosaccharide which may be selected from C6 or C5 sugars, sugar acids, and amino sugars. Embodiment 9. The C6 or C5 sugar is selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, growth, idose, talose, aldose, and ketose, The sugar acid is selected from gluconic acid, aldonic acid, uronic acid, and urosonic acid, or The amino sugar is selected from glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine. The linker compound according to embodiment 8. Appearance 10 The linker compound according to any one of embodiments 1 to 9, wherein the β-position is formed from a functional group of a precursor compound with a polar group, and the functional group is selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctin, hydrazine, carbonylalkyl heteroaryl, and their protected forms. Appearance 11 .below: (a) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -R 30(XXX) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene group. R 30 C3~C may be substituted. 10 Selected from carbocyclic, thiourea, optionally substituted thiourea, urea, optionally substituted urea, sulfamide, alkylsulfamide, acylsulfamide, optionally substituted alkylsulfamide, optionally substituted acylsulfamide, sulfonamide, optionally substituted sulfonamide, guanidine containing alkyl and arylguanidine, phosphoramide, or optionally substituted phosphoramide; or R 30 These include azide, alkynyl, substituted alkynyl, -NH-C(O)-alkynyl, and -NH-C(O)-alkynyl-R 65 Selected from cyclooctin, -NH-cyclooctin, -NH-C(O)-cyclooctin, or -NH-(cyclooctin)2, where R 65 This is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbon ring, optionally substituted aryl, optionally substituted heterocarbon ring, or optionally substituted heteroaryl. n20 is between 2 and 26; (b) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NH-C(O)-R 31 (XXXI) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21and R 22 Each of these is independently bonded or a C1-C3 alkylene group. R 31 It is a branched polyethylene glycol chain, where each branch has 1 to 26 ethylene glycol subunits and each branch has R at its end. 35 It has, R 35 azid, alkinyl, alkinyl-R 65 , cyclooctin or cyclooctin-R 65 And here, R 65 This is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbon ring, optionally substituted aryl, optionally substituted heterocarbon ring, or optionally substituted heteroaryl. n20 is between 2 and 26; (c) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -C(O)NH-R 31 (XXXII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene group. R 31 This is a branched polyethylene glycol chain, where each branch independently has 1 to 26 ethylene glycol subunits, and each branch has R at its end. 35 It has, R 35 azid, alkinyl, alkinyl-R 65 , cyclooctin or cyclooctin-R 65 And here, R 65This is selected from optionally substituted alkyls, optionally substituted alkenyls, optionally substituted alkynyls, optionally substituted carbon rings, optionally substituted aryls, optionally substituted heterocarbon rings, and optionally substituted heteroaryls. n20 is between 2 and 26; (d) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -C(O)NR 31 -R 22 -NR 24 R 25 (XXXIII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 31 is H or R 22 -NR 24 R 25 And, R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene group. R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, and a substituted -C(O)-polyhydroxyl group, however, R 24 and R 25 It is not possible for both to be H. n20 is between 2 and 26; (e) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -N(R 33 -R 31 )2(XXXIV) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene group. R 31 It is a branched polyethylene glycol chain, where each branch has 1 to 26 ethylene glycol subunits and each branch has R at its end. 35 It has, R 33 These are C1~C3 alkylene, C1~C3 alkylene-C(O), -C(O)-C1~C3 alkylene, or -C(O)-C1~C3 alkylene-C(O), R 35 azid, alkinyl, alkinyl-R 65 , cyclooctin or cyclooctin-R 65 And here, R 65 This is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbon ring, optionally substituted aryl, optionally substituted heterocarbon ring, or optionally substituted heteroaryl. n20 is between 2 and 26; (f) ~R 20 -(R 21 -[CH2-CH(OR 34 )-CH2-O] n20 -R 36 ) n25 (XXXV) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. Each R 21 These are independently a bond, an -O- or a C1-C3 alkylene group, Each R 34 These are independently H, -[CH2-CH(OH)-CH2-O] n20 -R 36 -C(O)-NR 24 R 25 or -C(O)N(R N )-C1~C6 Alkylene-NR 24 R25 And, R N is H or C1-C4 alkyl, R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, however, R 24 and R 25 It is not possible for both to be H. Each R 36 These are independently H, C1~C6 alkylene-C(OH)H-NR 44 R 45 C1~C6 alkylene-C(OH)H-C1~C6 alkylene-NR 44 R 45 -C(O)-NR 24 R 25 ,-C(O)N(R N )-C1~C6 Alkylene-NR 24 R 25 C1-C6 alkylene-C(O)NR 24 R 25 Or C1~C6 alkylene-CO2R 37 And, Each R 37 These are independently H or C1-C6 alkyl groups. R 44 and R 45 Each of these is independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, and a substituted -C(O)-polyhydroxyl group. However, R 44 and R 45 It is not possible for both to be H. , Each n20 is independently between 1 and 26. n25 is either 1 or 2; (g) ~R 20 -R 21 -[[CH2-CH2-O] n20 -R 22 -[CH2-[CH(OH)] n23 -CH2-O] n21 ] n22 -R 23 -NR 24 -R25 (XXXVI) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 , R 22 and R 23 Each of these is independently bonded or a C1-C3 alkylene group. R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, and a substituted -C(O)-polyhydroxyl group, however, R 24 and R 25 It is not possible for both to be H. Each n20 is independently between 0 and 26, and each n21 is independently between 0 and 26, provided that at least one of n20 or n21 is between 2 and 26. n22 is 1 to 5. Each n23 is independently either 1 or 2; (h) ~R 20 -(R 21 -[O-CH2-CH2] n20 -R 22 -N(R N )-CO2-[CH2-CH(OR 34 )-CH2-O] n21 -R 36 ) n25 (XXXVII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene group. R N is H or C1-C4 alkyl, R 24 and R 25Each of these is independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, however, R 24 and R 25 It is not possible for both to be H. Each R 34 These are independently H, -[CH2-CH(OH)-CH2-O] n20 -R 36 , or -C(O)N(R N )-C1~C6 Alkylene-NR 24 R 25 And, Each R 36 These are independently H, C1~C6 alkylene-C(OH)H-NR 44 R 45 C1~C6 alkylene-C(OH)H-C1~C6 alkylene-NR 44 R 45 ,-C(O)N(R N )-C1~C6 Alkylene-NR 24 R 25 C1-C6 alkylene-C(O)NR 24 R 25 Or C1~C6 alkylene-CO2R 37 And, Each R 37 These are independently H or C1-C6 alkyl groups. R 44 and R 45 Each of these is independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, and a substituted -C(O)-polyhydroxyl group, however, R 44 and R 45 It is not possible for both to be H. n20 is between 2 and 26. n21 is between 1 and 26. n25 is either 1 or 2; (i) ~R 20 -(R 21 -[N(R N )-C(O)-[O-CH2-CH(OH)-CH2] n20 ] n21 -R 22 -NR 24 R 25 )n25 (XXXVIII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene group. R N is H or C1-C4 alkyl, R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, however, R 24 and R 25 It is not possible for both to be H. n20 is between 2 and 26. n21 is 1 to 4. n25 is 1, 2, or 3; (j) ~R 20 -(R 21 -[C(R α )HC(O)-N(R N )] n20 -R 22 -[CH2-CH2-O] n20 -NR 24 R 25 ) n25 (XXXIX) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently linked, C1~C3 alkylene, -C1~C3 alkylene-[O-CH2-CH2-] n20 ,-[CH2-CH2-O] n20 -C1~C3 alkylene- or -C1~C3 alkylene-[O-CH2-CH2-] n20 -C(O)-, Each R α H or -R are independent.22 -NR 24 R 25 And, Each R N These are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 And, R 24 and R 25 Each of these independently represents H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and an optional C3-C3 group. 10 Carbon rings, unsubstituted C1-C3 alkylenes, C3-C 10 Carbocyclic ring, optionally substituted heteroaryl, optionally substituted carbocyclic ring, substituted-C1~C8 alkyl, substituted-C(O)-C1~C8 alkyl, chelating agent, R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More selected; or -NR 24 R 25 They combine to form a C3-C8 heterocycle, however, R 24 and R 25 It is not possible for both to be H. Each n20 is independently between 0 and 26, except that at least one n20 is between 2 and 26. n25 is 1 or 2; or (k) TIFF2026534278000010.tif25128 or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 , R 22 and R 23 Each of these is independently linked, C1~C3 alkylene, -C1~C3 alkylene-[O-CH2-CH2-] n20 ,-[CH2-CH2-O] n20 -C1~C3 alkylene- or -C1~C3 alkylene-[O-CH2-CH2-] n20-C(O)-, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N These are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 And, R 24 and R 25 Each of these independently represents H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and an optional C3-C3 group. 10 Carbon rings, unsubstituted C1-C3 alkylenes, C3-C 10 Carbocyclic ring, optionally substituted heteroaryl, optionally substituted carbocyclic ring, substituted-C1~C8 alkyl, substituted-C(O)-C1~C8 alkyl, chelating agent, R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More selected; or -NR 24 R 25 They combine to form a C3-C8 heterocycle, however, R 24 and R 25 It is not possible for both to be H. R 26 is H or C1-C6 alkyl, Each n20 is independently between 0 and 26, except that at least one n20 is between 2 and 26. Each n21 is independently between 0 and 26, except that at least one n21 is between 2 and 26; A linker compound according to any one of embodiments 1 to 10, comprising a polar group having a more selectable formula. Appearance 12 .below: ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NH-C(O)-R 31 (XXXI) ~R 20 -R21 -[O-CH2-CH2] n20 -R 22 -C(O)NH-R 31 (XXXII), and ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -N-(R 33 -R 31 )2(XXXIII), A polar group comprising a more selected formula or its stereoisomer or salt, During the ceremony, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each of these is independently bonded or a C1-C3 alkylene group. R 31 It is a branched polyethylene glycol chain, where each branch has 1 to 26 ethylene glycol subunits and each branch has R at its end. 35 It has, R 33 These are C1~C3 alkylene, -C1~C3 alkylene-C(O), -C(O)-C1~C3 alkylene, or -C(O)-C1~C3 alkylene-C(O), R 35 azid, alkinyl, alkinyl-R 65 , cyclooctin or cyclooctin-R 65 And here, R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbon ring, optionally substituted aryl, optionally substituted heterocarbon ring, or optionally substituted heteroaryl, and the wavy (~) line is R 20 This shows the binding site, and n20 is 2 to 26. A linker compound according to any one of embodiments 1 to 11. Appearance 13 . To site β, site R bA linker compound according to any one of embodiments 11 to 12, wherein the bond to or to an enzymatically cleavable group is formed from a functional group of a polar group precursor compound, and the functional group is selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctin, hydrazine, carbonylalkyl heteroaryl, and their protected forms. Appearance 14 .formula: ~R 20 -(R 43 -R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XL) or a polar group having a stereoisomer or salt thereof, During the ceremony, R 20 This is directed to site β, and site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 41 and R 42 Each of these is independently bonded or a C1-C6 alkylene. Each R 43 These are either independently bonded or C1~C 12 Alkylene, -NH-C1~C 12 Alkylene, -C1~C 12 Alkylene-NH-,-C(O)-C1~C 12 Alkylene, -C1~C 12 Alkylene-C(O)-,-NH-C1~C 12 Alkylene -C(O)-, -C(O)-C1~C 12 Alkylene-NH-, -NH-C(O)-NH-, -NH-C(O)-, -NH-C(O)-C1~C 12 Alkylene, -C(O)-NH-C1~C 12 Alkylene, -heteroarylene, heteroaryl-C1~C12 Alkylene, heteroaryl-C1~C 12 Alkylene-C(O)- or -C(O)NR 46 R 47 Selected from, here, R 46 and R 47 One of them is H or C1~C 12 It is an alkylene, and the other is C1~C 12 It is alkylene, R 44 and R 45 Each of these is independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituent is selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and n40 is 2 to 26. n41 is 1 to 6. n42 is 1 to 6. A linker compound according to any one of embodiments 1 to 13. Appearance 15 .formula: ~R 20 -(R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLI) or a polar group having a stereoisomer or salt thereof, During the ceremony, R 20 This is directed to site β, and site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 41 and R 42 Each of these is independently bonded or a C1-C6 alkylene. R 43 is a bond, or C1~C 12 Alkylene, -NH-C1~C 12 Alkylene, -C1~C 12 Alkylene-NH-,-C(O)-C1~C 12Alkylene, -C1~C 12 Alkylene-C(O)-,-NH-C1~C 12 Alkylene -C(O)-, -C(O)-C1~C 12 Alkylene-NH-, -NH-C(O)-NH-, -NH-C(O)-, -NH-C(O)-C1~C 12 Alkylene, C(O)-NH-C1~C 12 Alkylene, -heteroarylene, heteroaryl-C1~C 12 Alkylene, heteroaryl-C1~C 12 Alkylene-C(O)- or -C(O)NR 46 R 47 Selected from, here, R 46 and R 47 One of them is H or C1~C 12 It is an alkylene, and the other is C1~C 12 It is alkylene, R 44 and R 45 Each of these is independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituent is selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and n40 is 1 to 26. n41 is 1 to 6. n42 is 1 to 6. A linker compound according to any one of embodiments 1 to 14. Appearance 16 .formula: ~R 20 -(R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLII) or a polar group having a stereoisomer or salt thereof, During the ceremony, R 20 This is directed to site β, and site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 41 and R 42 Each of these is independently bonded or a C1-C3 alkylene. R 43 This is either a bond or a C1-C6 alkylene, -NH-C1-C 12 Alkylene, -C1~C6 alkylene-NH-, -C(O)-C1~C6 alkylene, -C1~C6 alkylene-C(O)-, -NH-C1~C6 alkylene-C(O)-, -C(O)-C1~C6 alkylene-NH-, -NH-C(O)-NH-, -NH-C(O)-, -NH-C(O)-C1~C6 alkylene, -C(O)-NH-C1~C 12 Alkylene, -heteroarylene, heteroaryl-C1~C6 alkylene, heteroaryl-C1~C6 alkylene-C(O)-, or -C(O)NR 46 R 47 Selected from, here, R 46 and R 47 One of them is H or C1-C6 alkylene, and the other is C1-C 12 It is alkylene, R 44 and R 45 Each of these is independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituent is selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and n40 is 1 to 16. n41 is 1 to 4. n42 is 1-4. A linker compound according to any one of embodiments 1 to 15. Appearance 17 .R 20The linker compound according to any one of embodiments 6, 11, 12 and 14-16, wherein the linker is formed from a functional group of a polar group precursor compound, and the functional group is selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctin, hydrazine, carbonylalkyl heteroaryl, or a protected form thereof. Appearance 18 .R 20 However, the structure is as follows: It includes one of TIFF2026534278000011.tif68141, TIFF2026534278000012.tif202143, or TIFF2026534278000013.tif28145 or its stereoisomer, During the ceremony, R is H, C1-C6 alkyl, or polyhydroxyl group. n is between 0 and 12. TIFF2026534278000014.tif4128 is for site β or site R b It shows binding to or to a group that can be enzymatically cleaved, TIFF2026534278000015.tif4128 shows the binding site to the rest of the polar group. A linker compound according to any one of embodiments 6, 11, 12, and 14-16. Appearance 19 .R 20 However, the structure is as follows: Having one of TIFF2026534278000016.tif158150 or TIFF2026534278000017.tif158149 or a stereoisomer thereof, During the ceremony, n is between 0 and 12. TIFF2026534278000018.tif4128 is for site β or site R b It shows binding to or to a group that can be enzymatically cleaved, TIFF2026534278000019.tif4128 shows the binding site to the rest of the polar group. The compound according to any one of embodiments 6, 11, 12, and 14-16. Appearance 20 .R 43 -(NR 44 R 45 ) n41 However, the structure is as follows: Having one of TIFF2026534278000020.tif123137 or its stereoisomer, During the ceremony, R is H, C 1~6 Alkyl, polyhydroxyl, or substituted polyhydroxyl groups, TIFF2026534278000021.tif4128 is R to the rest of the polar group 43 The binding site is shown. A linker compound according to any one of embodiments 14 to 19. Appearance 21 .R 43 -(NR 44 R 45 ) n41 However, the structure is as follows: Having one of TIFF2026534278000022.tif68131 or TIFF2026534278000023.tif76145 or a stereoisomer thereof, During the ceremony, TIFF2026534278000024.tif4128 is R to the rest of the polar group 43 The binding site is shown. A linker compound according to any one of embodiments 14 to 16. Appearance 22 .-NR 44 R 45 However, the structure is as follows: Having one of TIFF2026534278000025.tif109136 or TIFF2026534278000026.tif166150 or its stereoisomer, During the ceremony, TIFF2026534278000027.tif4128 is -NR to the rest of the polar group 44 R45 The binding site is shown. A linker compound according to any one of embodiments 14 to 21. Appearance 23 The following structure precedes the linker unit: TIFF2026534278000028.tif213142TIFF2026534278000029.tif21994TIFF2026534278000030.tif203105TIFF20 26534278000031.tif183147TIFF2026534278000032.tif204149TIFF2026534278000033.tif178149TIFF20265342 78000034.tif201153TIFF2026534278000035.tif143147TIFF2026534278000036.tif196149TIFF2026534278000 037.tif193142TIFF2026534278000038.tif177148TIFF2026534278000039.tif193156TIFF2026534278000040.ti f198161TIFF2026534278000041.tif222147TIFF2026534278000042.tif186154TIFF2026534278000043.tif1861 52TIFF2026534278000044.tif169150TIFF2026534278000045.tif187133TIFF2026534278000046.tif204152TIFF It contains a polar group having one of the following: 2026534278000047.tif213154TIFF2026534278000048.tif22897TIFF2026534278000049.tif187128TIFF2026534278000050.tif226156TIFF2026534278000051.tif190159TIFF2026534278000052.tif171155 During the ceremony, (*) indicates β or site R b It indicates a binding site to or to a group that can be enzymatically cleaved, Each R is independently H or C1-C6 alkyl. R' is H, C1-C6 alkyl, -N(R 24 )(R 25 ) or -CO2H, Each n is independently between 1 and 12. X is O, NR, or -CH2-, V is a bond or a C1-C6 alkyl group. R 24 and R 25 One of them is H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, substituted -C(O)-C1~C8 alkyl, chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More selected, R 24 and R 25 The other side consists of H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, substituted -C(O)-C1~C8 alkyl, chelating agent, R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 , and polyethylene glycol which may have 1 to 24 ethylene glycol subunits, however, R 24 and R 25 Both cannot be H. A linker compound according to any one of embodiments 1 to 22. Pattern 24 .below: TIFF2026534278000053.tif119145TIFF2026534278000054.tif217144TIFF2026534278000055.tif229144TIFF2026534278000056.tif20 3145TIFF2026534278000057.tif188144TIFF2026534278000058.tif164141TIFF2026534278000059.tif197143TIFF2026534278000060.t if223144TIFF2026534278000061.tif211144TIFF2026534278000062.tif220145TIFF2026534278000063.tif201145TIFF20265342780000 64.tif182144TIFF2026534278000065.tif203144TIFF2026534278000066.tif207144TIFF2026534278000067.tif213135TIFF20265342780 00068.tif208134TIFF2026534278000069.tif221134TIFF2026534278000070.tif221132TIFF2026534278000071.tif193136TIFF2026534 278000072.tif211126TIFF2026534278000073.tif188133TIFF2026534278000074.tif215110TIFF2026534278000075.tif20373TIFF2026 534278000076.tif219144TIFF2026534278000077.tif174144TIFF2026534278000078.tif214146TIFF2026534278000079.tif176145TIFF2026534278000080.tif192145TIFF2026534278000081.tif193144TIFF2026534278000082.tif104133 or a polar group selected from their stereoisomers or salts, During the ceremony, Each Z is connected at *, independently Selected from TIFF2026534278000083.tif113145, each TIFF2026534278000084.tif2128 is a site R to site β. b This is a bond to, or to, a group that can be enzymatically cleaved. A linker compound according to any one of embodiments 1 to 23. Appearance 25 A linker compound according to any one of embodiments 1 to 24, comprising a polar group containing polymer units and sugar units. Appearance 26 A linker compound according to any one of embodiments 1 to 24, comprising a polar group containing at least two polymer units. Appearance 27 A linker compound according to any one of embodiments 1 to 24, comprising a polar group containing polymer units and carboxyl units. Appearance 28 A linker compound according to any one of embodiments 1 to 24, comprising at least two polar groups. Appearance 29 A linker compound according to any one of embodiments 1 to 24, comprising a polar group containing polymer units, sugar units, and carboxyl units. Appearance 30 A linker compound according to any one of embodiments 1 to 24, comprising a polar group having at least two polymer units, at least one sugar unit, and at least one carboxyl unit. Appearance 31 A linker compound according to any one of embodiments 1 to 24, wherein the enzyme-cleavable group comprises at least two amino acids. Appearance 32 A linker compound according to any one of embodiments 1 to 31, comprising at least one polar group bonded to an enzymatically cleavable group. Appearance 33 The following structure: It has one of the following: TIFF2026534278000085.tif35128TIFF2026534278000086.tif192145TIFF2026534278000087.tif187133TIFF2026534278000088.tif46128 During the ceremony, R c is a combination or C 1~6 It is alkylene, The wavy line on the amino group indicates a binding site for the stretcher group, or indicates H before binding to the stretcher group. β- is a binding site to at least one polar group, The benzyl H on the benzyl OH may be replaced by a bond to at least one of the drug units or to a linking group bonded to at least one of the drug units. A linker compound according to any one of embodiments 1 to 32. Appearance 34 A linker compound according to any one of embodiments 1 to 33, wherein the enzyme-cleavable group contains a peptide that can be cleaved by an intracellular protease. Embodiment 35. The linker compound according to Embodiment 34, wherein the intracellular protease is cathepsin B. Appearance 36 The linker compound according to embodiment 34, wherein the enzymatically cleavable group comprises a cleavable peptide including valine-citrulline peptide, valine-alanine peptide, valine-lysine peptide, phenylalanine-lysine peptide, or glycine-glycine-phenylalanine-glycine peptide. Appearance 37 The following structure: It includes one of the following: TIFF2026534278000089.tif198125TIFF2026534278000090.tif153128TIFF2026534278000091.tif202103TIFF2026534278000092.tif205107TIFF2026534278000093.tif184128TIFF2026534278000094.tif155128TIFF2026534278000095.tif224102TIFF2026534278000096.tif68128 During the ceremony, R C is a combination or C 1~6 It is alkylene, The wavy line on the amino group indicates a binding site for the stretcher group, or indicates H before binding to the stretcher group. β- is the binding site to the POLY unit. The hydrogen atoms on the benzyl OH group may be replaced by binding to at least one drug unit, or to a binding site to at least one drug unit. A linker compound according to any one of embodiments 1 to 36. Appearance 38 The following structure: TIFF2026534278000097.tif63143TIFF2026534278000098.tif222145TIFF2026534278000099.tif1881 44TIFF2026534278000100.tif189144TIFF2026534278000101.tif204144TIFF2026534278000102.tif20 4145TIFF2026534278000103.tif219144TIFF2026534278000104.tif203144TIFF2026534278000105.tif 213144TIFF2026534278000106.tif194145TIFF2026534278000107.tif174145TIFF2026534278000108.t if193144TIFF2026534278000109.tif192144TIFF2026534278000110.tif220144TIFF202653427800011 1.tif232145TIFF2026534278000112.tif208145TIFF2026534278000113.tif224145TIFF2026534278000 It has one of the following structures: 114.tif208145TIFF2026534278000115.tif141143TIFF2026534278000116.tif192145TIFF2026534278000117.tif192145TIFF2026534278000118.tif162144TIFF2026534278000119.tif222144 During the ceremony, The wavy line on the amino group indicates the binding site to the stretcher group, or indicates H before binding to the stretcher group. The hydrogen atoms on the benzyl OH group may be replaced by bonds to at least one drug unit, or to a linking group bonded to at least one drug unit. A linker compound according to any one of embodiments 1 to 4. Appearance 39 A linker compound according to any one of embodiments 1 to 36, wherein an enzyme-cleavable group is attached to a stretch group by a non-peptide linking group. Pattern 40 The non-peptide linking group may be substituted C1-C 10 Alkylene, C2-C may be substituted. 10 Alkenylene, C2-C2 may be substituted. 10 A linker compound according to embodiment 39, selected from alkylylene or optionally substituted polyethylene glycol. Appearance 41 A linker compound according to any one of embodiments 1 to 40, comprising a stretcher group bonded to an enzymatically cleavable group. Pattern 42 The stretcher group is as follows: Selected from TIFF2026534278000120.tif83137, During the ceremony, R 17 is -C1~C 10 Alkylene-, -C1~C 10 Heteroalkylene-, -C3~C8 carbocyclo-, -O-(C1~C8 alkylene)-, -(CH2-O-CH2) b -C1~C8 alkylene-(where b is 1~26), -C1~C8 alkylene-(CH2-O-CH2) b -(where b is 1 to 26), -C1~C8 alkylene-(CH2-O-CH2) b -C1~C8 Alkylene- (where b is 1~26), -Arirene-, -C1~C 10 Alkilen-Arirene-, -Arirene-C1~C 10 Alkylene-, -C1~C 10Alkylene-(C3~C8 carbocyclo)-,-(C3~C8 carbocyclo)-C1~C 10 Alkylene-, -C3~C8 heterocyclo-, -C1~C 10 Alkylene-(C3~C8 heterocyclo)-,-(C3~C8 heterocyclo)-C1~C 10 Alkylene-, -C1~C 10 Alkylene -C(=O)-, -C1~C 10 Alkylene-C(O)NH-C1~C8 Alkylene-[O-CH2-CH2] n -C(O)- (where n is 1 to 26), C1~C 10 Heteroalkylenes -C(=O)-, -C1~C8 alkylenes-(CH2-O-CH2) b -C(=O)-(where b is 1 to 26), -(CH2-O-CH2) b -C1~C8 alkylene-C(=O)-(where b is 1~26), -C1~C8 alkylene-(CH2-O-CH2) b -C1~C8 alkylene-C(=O)- (where b is 1~26), -C3~C8 carbocyclo-C(=O)-, -O-(C1~C8 alkyl)-C(=O)-, -arylene-C(=O)-, -C1~C 10 Alkylene-arylene-C(=O)-,-arylene-C1~C 10 Alkylene -C(=O)-, -C1~C 10 Alkylene-(C3~C8 carbocyclo)-C(=O)-,-(C3~C8 carbocyclo)-C1~C 10 Alkylene-C(=O)-, -C3~C8 heterocyclo-C(=O)-, -C1~C 10 Alkylene-(C3~C8 heterocyclo)-C(=O)-,-(C3~C8 heterocyclo)-C1~C 10 Alkylene -C(=O)-, -C1~C 10 Alkylene-NH-,-C1~C 10 Heteroalkylene-NH-,-C1~C8 alkylene-(CH2-O-CH2) b -NH-(where b is 1 to 26), -(CH2-O-CH2) b-C1~C8 alkylene-NH- (where b is 1~26), -C1~C8 alkylene-(CH2-O-CH2) b -C1~C8 alkylene-NH- (where b is 1~26), -C1~C8 alkylene-(C(=O))-NH-(CH2-O-CH2) b -C(=O)-(where b is 1 to 26), -C1~C8 alkylene-(C(=O))-NH-(CH2-O-CH2) b -C1~C8 alkylene-C(=O)-(where b is 1~26), -C1~C8 alkylene-NH-(C(=O))-(CH2-O-CH2) b -NH- (where b is 1 to 26), -C1~C8 alkylene-NH-(C(=O))-(CH2-O-CH2) b -C1~C8 alkylene-NH- (where b is 1~26), -C3~C8 carbocyclo-NH-, -O-(C1~C8 alkyl)-NH-, -arylene-NH-, -C1~C 10 Alkylene-arylene-NH-,-arylene-C1~C 10 Alkylene-NH-,-C1~C 10 Alkylene-(C3~C8 carbocyclo)-NH-,-(C3~C8 carbocyclo)-C1~C 10 Alkylene-NH-,-C3~C8 heterocyclo-NH-,-C1~C 10 Alkylene-(C3~C8 heterocyclo)-NH-,-(C3~C8 heterocyclo)-C1~C 10 Alkylene-NH-,-C1~C 10 Alkylene-S-, C1~C 10 Heteroalkylene-S-, -C3~C8 carbocyclo-S-, -O-(C1~C8 alkyl)-S-, -arylene-S-, -C1~C 10 Alkylene-Arirene-S-,-Arirene-C1~C 10 Alkylene-S-,-C1~C 10 Alkylene-(C3~C8 carbocyclo)-S-,-(C3~C8 carbocyclo)-C1~C 10 Alkylene-S-, -C3~C8 heterocyclo-S-, -C1~C 10Alkylene-(C3~C8 heterocyclo)-S-, or-(C3~C8 heterocyclo)-C1~C 10 It is alkylene-S-, or The stretcher group is maleimide (C1~C 10 Alkylene-C(O)-, maleimide (CH2OCH2) p2 (C1~C 10 Alkylene)C(O)-, Maleimide (C1~C 10 Alkylene (CH2OCH2) p2 This includes C(O)- or their ring-opened forms, where p2 is 1 to 26. * indicates binding to a PTK7-binding substance, and the wavy line indicates binding to a group that can be enzymatically cleaved. The linker compound according to embodiment 41. Appearance 43 The stretcher group is as follows: Selected from TIFF2026534278000121.tif193140, During the ceremony, wavy line TIFF2026534278000122.tif2128 shows the binding site of a stretcher group to an enzymatically cleavable group. The binding site to the PTK7 binding material is located on the maleimide, primary amine, or alkyne functional group. The linker compound according to embodiment 41. 44 The following structure: TIFF2026534278000123.tif196145TIFF2026534278000124.tif190145TIFF2026534278000125.ti f224145TIFF2026534278000126.tif175144TIFF2026534278000127.tif229144TIFF2026534278000 128.tif224144TIFF2026534278000129.tif181145TIFF2026534278000130.tif184145TIFF202653 4278000131.tif204145TIFF2026534278000132.tif168145TIFF2026534278000133.tif225145TIFF 2026534278000134.tif208145TIFF2026534278000135.tif188145TIFF2026534278000136.tif181 145TIFF2026534278000137.tif225145TIFF2026534278000138.tif213144TIFF2026534278000139. It has one of the following: tif219145TIFF2026534278000140.tif188145TIFF2026534278000141.tif229144TIFF2026534278000142.tif194144TIFF2026534278000143.tif207145TIFF2026534278000144.tif64144 In the formula, the hydrogen atoms on the benzyl OH group may be replaced by bonds to at least one drug unit or to a linking group bonded to at least one drug unit. The linker compound according to Embodiment 1. Appearance 45 The linker compound according to any one of embodiments 1 to 44, wherein at the binding site δ, it is bound to at least one drug unit or to a linking group bound to at least one drug unit. A drug-linker compound containing such compounds. Appearance 46A drug-linker according to embodiment 45, wherein the drug unit is selected from cytotoxic substances, immunomodulators, nucleic acids, growth inhibitors, PROTACs, toxins, radioisotopes, and chelate ligands. Appearance 47 A drug-linker according to embodiment 46, wherein the drug unit is a cytotoxic substance. 48 A drug-linker according to embodiment 47, wherein the cytotoxic substance is selected from the group consisting of auristatin, meitansinoid, camptothecin, duocalmycin, and calicheamycin. Appearance 49 A drug-linker according to embodiment 48, wherein the cytotoxic substance is auristatin. Appearance 50 A drug-linker according to embodiment 48, wherein the cytotoxic substance is camptothecin. Appearance 51 A drug-linker according to embodiment 50, wherein the cytotoxic substance is RS-exatecan or SS-exatecan. Appearance 52 A drug-linker according to embodiment 51, wherein the meitansinoid is meitansin, meitansinol, or ansamatosin-2. Appearance 53 A drug-linker according to embodiment 46, wherein the drug unit is an immunomodulator. Phenomenon 54 A drug-linker according to embodiment 53, wherein the immunomodulator is selected from a TRL7 agonist, a TLR8 agonist, a STING agonist, or a RIG-I agonist. Appearance 55 A drug-linker according to embodiment 54, wherein the immunomodulator is a TLR7 agonist. Appearance 56The drug-linker according to embodiment 55, wherein the TLR7 agonist is imidazoquinoline, imidazoquinolineamine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzoimidazole-2-amine, tetrahydropyridopyrimidine, heteroalothiadiazide-2,2-dioxide, benzonaphthyridine, guanosine analog, adenosine analog, thymidine homopolymer, ssRNA, CpG-A, polyG10, or polyG3. Appearance 57 A drug-linker according to embodiment 54, wherein the immunomodulator is a TLR8 agonist. Appearance 58 A drug-linker according to embodiment 57, wherein the .TLR8 agonist is selected from imidazoquinoline, thiazoloquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzoimidazole-2-amine, tetrahydropyridopyrimidine, or ssRNA. Appearance 59 A drug-linker according to embodiment 53, wherein the immunomodulator is a STING agonist. Appearance 60 A drug-linker according to embodiment 54, wherein the immunomodulator is a RIG-I agonist. Appearance 61 A drug-linker according to embodiment 60, wherein the .RIG-I agonist is selected from KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400 and KIN2000. Appearance 62 A drug-linker according to embodiment 46, wherein the drug unit is a chelate ligand. Appearance 63A drug-linker according to embodiment 62, wherein the chelate ligand is selected from platinum (Pt), ruthenium (Ru), rhodium (Rh), gold (Au), silver (Ag), copper (Cu), molybdenum (Mo), titanium (Ti), or iridium (iridum) (Ir); or from radioactive isotopes, e.g., yttrium-88, yttrium-90, technetium-99, copper-67, rhenium-188, rhenium-186, gallium-66, gallium-67, indium-111, indium-114, indium-115, lutetium-177, strontium-89, samarium-153, and lead-212. Appearance 64 The following structure: TIFF2026534278000145.tif81145TIFF2026534278000146.tif193145TIFF2026534278000147.tif212148TIFF 2026534278000148.tif219145TIFF2026534278000149.tif178128TIFF2026534278000150.tif179144TIFF202 6534278000151.tif178145TIFF2026534278000152.tif162145TIFF2026534278000153.tif208144TIFF202653 4278000154.tif191145TIFF2026534278000155.tif192145TIFF2026534278000156.tif198145TIFF202653427 8000157.tif163144TIFF2026534278000158.tif225148TIFF2026534278000159.tif174145TIFF202653427800 0160.tif186145TIFF2026534278000161.tif198145TIFF2026534278000162.tif224145TIFF202653427800016 A drug-linker according to embodiment 45, having one of the following: 3.tif219145TIFF2026534278000164.tif207144TIFF2026534278000165.tif194144TIFF2026534278000166.tif214144TIFF2026534278000167.tif168144TIFF2026534278000168.tif63143. Appearance 65 .Average drug loading (p load The composite according to any aspect 64, wherein the number of ) is approximately 1 to approximately 8, approximately 2, approximately 4, approximately 6, approximately 8, approximately 10, approximately 12, approximately 14, approximately 16, approximately 3 to approximately 5, approximately 6 to approximately 8, or approximately 8 to approximately 16. Appearance 66 .below: TIFF2026534278000169.tif75144TIFF2026534278000170.tif187143TIFF2026534278000171.tif 193145TIFF2026534278000172.tif203142TIFF2026534278000173.tif183142TIFF2026534278000 174.tif225143TIFF2026534278000175.tif219145TIFF2026534278000176.tif175147TIFF202653 4278000177.tif210144TIFF2026534278000178.tif214145TIFF2026534278000179.tif229148TIFF 2026534278000180.tif210144TIFF2026534278000181.tif188141TIFF2026534278000182.tif182 145TIFF2026534278000183.tif209146TIFF2026534278000184.tif193143TIFF2026534278000185 Selected from .tif213146TIFF2026534278000186.tif188144TIFF2026534278000187.tif184147TIFF2026534278000188.tif215147TIFF2026534278000189.tif222151TIFF2026534278000190.tif127144, During the ceremony, Ab is a PTK7 binding substance, n is p load That is, The composite according to any one of embodiments 64 to 65. In some embodiments, the complex is a stereoisomer of one of the structures described above. Appearance 67 - Exemplary complex LD110 complex TIFF2026534278000191.tif66147LD163 complex TIFF2026534278000192.tif63151LD038 complex TIFF2026534278000193.tif60128LD343 complex In some embodiments, the complex is a stereoisomer of one of the structures described above. Appearance 68 - Exemplary complex LD343 complex TIFF2026534278000195.tif52143In formula, Ab is a binding substance comprising a heavy chain having a heavy chain variable (VH) region and a heavy chain steady region, and a light chain having a light chain variable (VL) region and a light chain steady region. The VH region has the amino acid sequence shown in SEQ ID NO:15. The VL region has the amino acid sequence shown in SEQ ID NO:16. The heavy chain constant region has the amino acid sequence shown in SEQ ID NO:56. The light chain constant region has the amino acid sequence shown in SEQ ID NO:57. n is p load And here, p load It is 8. In some embodiments, the complex is a stereoisomer of one of the structures described above.

[0245] Drug loading The complex may contain one or more drug units per PTK7-binding substance. The number of drug units per PTK7-binding substance is called the drug loading. The drug loading of a complex is the average number of drug units (drug molecules (e.g., cytotoxic substances)) per PTK7-binding substance (e.g., antibody or antigen-binding portion or non-antibody scaffold or non-antibody protein) in the complex, p load It is represented as p. load When p is approximately 4, the average drug load, considering all PTK7 binding substances present in the composition (e.g., antibody or antigen binding moieties or non-antibody scaffolds or non-antibody proteins), is approximately 4. In some embodiments, p loadThe range is approximately 3 to approximately 5, approximately 3.6 to approximately 4.4, or approximately 3.8 to approximately 4.2. In some embodiments, p load It may be about 3, about 4, or about 5. In some embodiments, p load The range is approximately 6 to approximately 8, more preferably approximately 7.5 to approximately 8.4. In some embodiments, p load The number may be approximately 6, approximately 7, or approximately 8. In some embodiments, p load The range is approximately 8 to 16.

[0246] The average number of drug units per PTK7-binding substance (e.g., antibody or antigen-binding moiety or non-antibody scaffold) in the preparation can be characterized by conventional methods such as UV, mass spectrometry, capillary electrophoresis (CE), and HPLC. load The quantitative distribution of the complex at this point can also be determined. In some cases, by means such as reverse-phase HPLC or hydrophobic interaction chromatography (HIC) HPLC, p load This method can achieve the separation, purification, and characterization of homogeneous complexes with a specific value from complexes with different drug loadings.

[0247] Binding of drug-linkers to antibodies, antigen-binding sites, and other binding materials (including non-antibody scaffolds) Techniques for conjugating PTK7-binding materials (such as antibodies or their antigen-binding moieties or non-antibody scaffolds) via linkers are well known in the art. See, for example, Alley et al., Current Opinion in Chemical Biology 2010 14:1-9; Senter, Cancer J., 2008, 14(3):154-169. In some embodiments, the linker is first conjugated to a drug unit (e.g., a cytotoxic substance, an immunomodulator, or other drug), and then the drug-linker is conjugated to the PTK7-binding material (e.g., an antibody or its antigen-binding moiety or non-antibody protein scaffold). In some embodiments, the linker is first conjugated to the PTK7-binding material (e.g., an antibody or its antigen-binding moiety or non-antibody protein scaffold), and then the drug unit is conjugated to the linker. In the following description, the term drug-linker is used to illustrate the conjugation of a linker or drug-linker to a PTK7-binding material, and those skilled in the art will recognize that the chosen conjugation method can be determined depending on the linker and drug unit. In some embodiments, the drug unit is bound to the PTK7 conjugate via a linker in such a manner that the activity of the drug unit is reduced until the drug unit is released from the complex (e.g., by hydrolysis, proteolysis, or by a cleavage agent).

[0248] In general, the complexes can be prepared by several routes employing organic chemical reactions, conditions, and reagents known to those skilled in the art, including (1) the reaction of a divalent linker with the nucleophile of a PTK7 conjugate (e.g., an antibody or its antigen-binding moiety or a non-antibody protein scaffold) to form a target-directing group-linker intermediate via covalent bonding, followed by reaction with a drug unit, and (2) the reaction of a divalent linker with the nucleophile of a drug unit to form a drug-linker via covalent bonding, followed by reaction with the nucleophile of a PTK7 conjugate. An exemplary method for preparing complexes via the latter route is described in U.S. Patent No. 7,498,298, which is incorporated herein by reference.

[0249] Nucleophiles on PTK7 conjugates, such as antibodies, antigen-binding sites, and other conjugates (including non-antibody scaffolds), non-limitingly include (i) N-terminal amine groups, (ii) side-chain amine groups, e.g., lysine, (iii) side-chain thiol groups, e.g., cysteine, and (iv) glycosylated sugar hydroxyl or amino groups. The amine, thiol, and hydroxyl groups are nucleophilic and can react with electrophiles on the linker to form covalent bonds, including (i) active esters such as NHS esters, HOBt esters, haloformates, and acid halides, (ii) alkyl and benzyl halides such as haloacetamides, and (iii) aldehydes, ketones, carboxyl, and maleimide groups. PTK7 conjugates, such as antibodies (and antigen-binding sites and other conjugates (including non-antibody scaffolds)), have reducible interchain disulfides, i.e., cysteine ​​crosslinks. Antibodies (and antigen-binding sites and other binding materials (including non-antibody scaffolds)) can be made more receptive to complexing with linkers by treatment with reducing agents such as DTT (dithiothreitol) or tricarbonylethylphosphine (TCEP) so that the antibody is completely or partially reduced. Thus, each cysteine ​​crosslink theoretically forms two reactive thiol nucleophiles. Additional nucleophiles can be introduced into PTK7 conjugates such as antibodies (and antigen-binding sites and other binding materials (including non-antibody scaffolds)) by modifying lysine residues, for example by reacting lysine residues with 2-iminothiolane (Trout's reagent) to convert the amine to a thiol. Reactive thiol groups can also be introduced into PTK7 conjugates (such as antibodies and antigen-binding sites and other binding materials (including non-antibody scaffolds)) by introducing one, two, three, or four or more cysteine ​​residues (for example, by preparing antibodies, antigen-binding sites, and other binding materials (including non-antibody scaffolds) containing one or more non-natural cysteine ​​amino acid residues).

[0250] The complex can also be generated by the reaction of an electrophile on a PTK7 binding material, such as an aldehyde or ketone carbonyl group, with a nucleophile on a linker reagent. Useful nucleophiles on the linker reagent include, but are not limited to, hydrazides, oximes, aminos, hydrazines, thiosemicarbazones, hydrazine carboxyls, and arylhydrazides. In one embodiment, the antibody (or its antigen-binding moiety or other binding material (including non-antibody scaffolds)) is modified to introduce an electrophilic moiety that can react with a nucleophilic substituent on the linker. In another embodiment, the sugar of the glycosylated antibody can be oxidized, for example, with a periodic acid oxidizing reagent to form an aldehyde or ketone group that can react with an amine group on the linker. The resulting imine-schiff base group can form a stable bond or can be reduced, for example, with a hydrogen boride reagent to form a stable amine bond. In one embodiment, the reaction of the carbohydrate portion of a glycosylated antibody with either galactose oxidase or sodium metaperiodate may yield a carbonyl (aldehyde and ketone) group in the antibody (or its antigen-binding portion, or other conjugate (including non-antibody scaffolds)) that can react with a suitable group on the linker (see, e.g., Hermanson, Bioconjugate Techniques). In another embodiment, a PTK7 conjugate, such as an antibody containing an N-terminal serine or threonine residue, can react with sodium metaperiodate to produce an aldehyde instead of the initial amino acid (Geoghegan & Stroh, (1992) Bioconjugate Chem. 3:138-146; U.S. Patent No. 5362852). Such an aldehyde can react with the linker.

[0251] Exemplary nucleophiles on drug units such as cytotoxic substances include, but are not limited to, amine, thiol, hydroxyl, hydrazide, oxime, hydrazine, thiosemicarbazone, hydrazine carboxyl, and arylhydrazide groups that can react to form a covalent bond with an electrophile on the linker, and include (i) active esters such as NHS esters, HOBt esters, haloformates, and acid halides, (ii) alkyl and benzyl halides such as haloacetamides, and (iii) aldehyde, ketone, carboxyl, and maleimide groups.

[0252] In some embodiments, the drug-linker is bound to an interchain cysteine ​​residue of the antibody (or its antigen-binding moiety or other binding material (including non-antibody scaffolds)). See, for example, International Publication No. 2004 / 010957 and International Publication No. 2005 / 081711. In such embodiments, the linker typically includes a maleimide group for the binding of an interchain disulfide to the cysteine ​​residue. In some embodiments, the linker or drug-linker is bound to a cysteine ​​residue of the antibody or its antigen-binding moiety, as described in U.S. Patent No. 7,585,491 or 8,080m250. The drug loading of the resulting complex is typically in the range of 1 to 8 or 1 to 16.

[0253] In some embodiments, the linker or drug-linker is conjugated to a lysine or cysteine ​​residue of the antibody (or its antigen-binding moiety or other binding substance) as described in International Publication No. 2005 / 037992 or International Publication No. 2010 / 141566. The drug loading of the resulting complex is typically in the range of 1 to 8.

[0254] In some embodiments, manipulated cysteine ​​residues, polyhistidine sequences, glycotechnology tags, or transglutaminase recognition sequences can be used for site-specific binding of linkers or drug-linkers to antibodies or their antigen-binding moieties or other binding materials (including non-antibody scaffolds).

[0255] In some embodiments, the drug-linker is attached to an engineered cysteine ​​residue at an Fc residue other than an interchain disulfide. In some embodiments, the drug-linker is attached to 118, 221, 224, 227, 228, 230, 231, 223 of the heavy chain of IgG (typically IgG1), according to Kabat's EU numbering. 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 275, 276, 278, 280, 281, 283, 285, 286, 291, 292, 293, 294, 295, 296, The manipulated cysteine ​​introduced at positions 297, 298, 299, 300, 302, 305, 313, 318, 323, 324, 325, 327, 328, 329, 330, 331, 332, 333, 335, 336, 396 and / or 428, as well as at positions 106, 108, 142(light chain), 149(light chain), and / or V205 of the light chain. An exemplary substitution relating to site-specific binding with the manipulated cysteine ​​is S239C (see, for example, US20100158909; Fc region numbering follows the EU index).

[0256] In some embodiments, the linker or drug-linker is conjugated to one or more introduced cysteine ​​residues of an antibody (or its antigen-binding moiety or other binding material (including non-antibody scaffolds)) as described in WO2006 / 034488, WO2011 / 156328 and / or WO2016040856.

[0257] In some embodiments, exemplary substitutions relating to site-specific binding using bacterial transglutaminase are N297S or N297Q in the Fc region. In some embodiments, the linker or drug-linker is conjugated to the glycan or modified glycan of the antibody or antigen-binding moiety or a glycosylated antibody (or other conjugating material, including non-antibody scaffolds). See, for example, WO2017 / 147542, WO2020 / 123425, WO2020 / 245229, WO2014 / 072482, WO2014 / 065661, WO2015 / 057066, and WO2016 / 022027, whose disclosures are incorporated herein by reference.

[0258] In some embodiments, a linker or drug-linker is bound to an antibody, antigen-binding moiety, or other binding material (including non-antibody scaffolds) via a saltase A linker. The saltase A linker can be produced by the saltase A enzyme fusing the LPXTG recognition motif (SEQ ID NO: 61) to the N-terminal GGG motif to regenerate a native amide bond.

[0259] In some embodiments, a linker or drug-linker is conjugated to an antibody, antigen-binding moiety, or other binding material (including non-antibody scaffolds) using SMARTag technology, where a bioorthogonal aldehyde handle is introduced to aldehyde-containing formylglycine (fGly) through oxidation of a cysteine ​​residue embedded in a specific peptide sequence (CxPxR). This enzymatic modification is carried out by formylglycinase (FGE). See, for example, Liu et al., Methods Mol. Biol. 2033:131-147 (2019).

[0260] In some embodiments, the linker or drug-linker is conjugated to an antibody, antigen-binding moiety, or other binding material (including non-antibody scaffolds) using cysteine ​​complexation with quaternary vinyl- and alkynyl-pyridine reagents. See, for example, Matos et al., Angew Chem. Int. Ed. Engl. 58:6640-6644 (2019).

[0261] In another embodiment, the linker or drug-linker is conjugated to an antibody, antigen-binding moiety, or other binding material (including a non-antibody scaffold) using a bismaleimide, C-lock, or K-lock method.

[0262] Pharmaceutical composition Other aspects of the complex relate to compositions containing an active ingredient, which include one of the complexes described herein. In some embodiments, the composition is a pharmaceutical composition. As used herein, the term “pharmaceutical composition” refers to an activator combined with a pharmaceutically acceptable carrier approved for use in the pharmaceutical industry. The expression “pharmaceutically acceptable” is used herein to mean a compound, substance, composition, carrier, and / or dosage form that, within the bounds of sound medical judgment, is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, and is balanced by a reasonable benefit-risk ratio.

[0263] The preparation of pharmacological compositions containing an active ingredient dissolved or dispersed therein is well understood in the art and does not need to be limited on any particular formulation. Typically, such compositions are prepared for injection as liquids or suspensions, however, solid forms suitable for rehydration to a liquid or suspension before use can also be prepared. Preparations may also be emulsified or provided as liposome compositions. The complex can be mixed with excipients that are pharmaceutically acceptable and have affinity for the active ingredient and are suitable for use in the therapeutics described herein. Suitable excipients include, for example, water, saline, dextrose, glycerol, ethanol, and combinations thereof. Furthermore, if desired, the pharmaceutical composition may contain small amounts of auxiliary substances, such as wetting or emulsifying agents, pH buffers, etc., that enhance or maintain the efficacy of the active ingredient (e.g., the complex).

[0264] The pharmaceutical compositions described herein may include pharmaceutically acceptable salts of their components. These pharmaceutically acceptable salts include, for example, acid addition salts formed with inorganic acids such as hydrochloric acid or phosphoric acid, or organic acids such as acetic acid, tartaric acid, or mandelic acid (formed with the free amino group of a polypeptide). Salts formed with the free carbone group can also be derived from inorganic bases such as sodium, potassium, ammonium, calcium, or ferric hydroxide, and organic bases such as isopropylamine, trimethylamine, 2-ethylaminoethanol, hisistazine, and prodecan.

[0265] Physiologically acceptable carriers are well known in the art. Exemplary liquid carriers are sterile aqueous solutions, such as phosphate-buffered saline, containing an active ingredient (e.g., a complex) and water, and may contain buffers such as sodium phosphate, saline, or both, at a physiological pH value. Furthermore, aqueous carriers may contain multiple buffer salts, salts such as sodium chloride and potassium chloride, dextrose, polyethylene glycol, and other solutes. Liquid compositions may also contain a liquid phase in addition to or excluding water. Examples of such additional liquid phases include glycerin, vegetable oils such as cottonseed oil, and water-oil emulsions. The amount of active ingredient effective in treating a particular disease or condition varies depending on the nature of the disease or condition and can be determined by standard clinical techniques.

[0266] In some embodiments, the pharmaceutical composition containing the complex may be a lyophilized agent.

[0267] In some embodiments, a syringe containing a therapeutically effective amount of the complex is provided.

[0268] Treatment method In some embodiments, a method is provided for treating a subject, comprising the step of administering the complex or pharmaceutical composition described herein to the subject. For example, in some embodiments, the subject has cancer or an autoimmune disease, and the complex binds to a target antigen associated with cancer or an autoimmune disease. Also provided are a complex or pharmaceutical composition described herein for use in treating a subject, comprising the step of administering the complex or pharmaceutical composition to the subject.

[0269] In some embodiments, a method for treating cancer is provided, comprising the step of administering a complex. In some embodiments, a subject requires treatment for cancer and / or malignant tumors. In some embodiments, the method is for treating a subject having cancer or malignant tumors.

[0270] The methods described herein include the step of administering a therapeutically effective dose of the complex to a subject having cancer or malignant tumor. As used herein, the terms “therapeutic dose,” “effective dose,” or “effective amount” refer to the amount of the complex that provides a therapeutic benefit in the treatment, management, or prevention of recurrence of cancer or malignant tumor, for example, the amount that provides a statistically significant reduction in at least one symptom, sign, or marker of the tumor or malignant tumor. Determining the therapeutically effective dose is well within the capabilities of those skilled in the art. Generally, the therapeutically effective dose may vary depending on the subject’s medical history, age, condition, sex, severity and type of the condition in the subject, and the administration of other pharmaceutically active agents.

[0271] The terms “cancer” and “malignant tumor” refer to the uncontrolled proliferation of cells that interferes with the normal functioning of organs and systems of the body. Cancer or malignant tumors can be primary or metastatic, meaning that cancer can invade and spread to tissues away from its original site. “Tumor” refers to the uncontrolled proliferation of cells that interferes with the normal functioning of organs and systems of the body. A subject with cancer is a subject that has objectively measurable cancer cells present in its body. This definition includes benign and malignant tumors, as well as potentially dormant tumors and micrometastases. When cancer moves from its original location and spreads to other vital organs, it can lead to the subject’s death through the decline of function in the affected organs. Hematological malignancies (hematopoietic cancers), such as leukemia and lymphoma, can, for example, overwhelm the normal hematopoietic compartments in a subject, leading to hematopoietic failure (in the form of anemia, thrombocytopenia, and neutropenia) and ultimately death.

[0272] Examples of cancer include, but are not limited to, carcinomas, lymphomas, germ tumors, sarcomas, and leukemias. More specific examples of such cancers include basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and CNS cancers, breast cancer (e.g., triple-negative breast cancer), peritoneal cancer, cervical cancer; bile duct cancer, choriocarcinoma, chondrosarcoma, colorectal cancer, connective tissue cancer, gastrointestinal cancer, endometrial cancer, esophageal cancer, eye cancer, head and neck cancer, gastrointestinal cancer (including gastrointestinal cancer and stomach cancer), glioblastoma (GBM), liver cancer, and hepatocellular carcinoma. Carcinoma in situ, renal cancer or kidney cancer (e.g., clear cell carcinoma), laryngeal cancer, leukemia, liver cancer, lung cancer (e.g., small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, and lung squamous cell carcinoma), lymphoma including Hodgkin lymphoma and non-Hodgkin lymphoma, melanoma, mesothelioma, myeloma, neuroblastoma, oral cancer (e.g., lip cancer, tongue cancer, oral cancer, and pharyngeal cancer), ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma , respiratory cancers, salivary gland cancers, sarcomas, skin cancers, squamous cell carcinomas, testicular cancers, thyroid cancers, endometrial or uterine cancers, intussusception cancers, urinary tract cancers, vulvar cancers; other carcinomas and sarcomas, B-cell lymphomas (low-grade / follicular non-Hodgkin lymphoma (NHL), small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small non-necrotic cell lymphomas) This includes, but is not limited to, NHL (including bulky NHL, mantle cell lymphoma, AIDS-associated lymphoma, and Waldenström macroglobulinemia), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myeloblastic leukemia, as well as post-transplant lymphoproliferative disorders (PTLD), facomatosis, edema (such as that associated with brain tumors), and abnormal angiogenesis associated with Meggs syndrome.

[0273] In some embodiments, PTK7+ cancer is a carcinoma. In some embodiments, the cancer is a squamous cell carcinoma.

[0274] In some embodiments, the cancer is a PTK7+ cancer. In some embodiments, the PTK7+ cancer is a solid tumor or a hematological malignancy.

[0275] In some embodiments, PTK7+ cancers are selected from breast cancer, ovarian cancer (OVCA), cervical cancer, pharyngeal cancer, gastric cancer, myeloma, bladder cancer, uterine cancer, esophageal squamous cell carcinoma (ESCC), colon cancer, hepatocellular carcinoma, and colorectal cancer.

[0276] In some embodiments, PTK7+ cancers are selected from esophageal squamous cell carcinoma (ESCC), lung cancer, and head and neck squamous cell carcinoma (HNSCC).

[0277] In some embodiments, the cancer is selected from bladder cancer, esophageal cancer, and uterine cancer.

[0278] In some embodiments, PTK7+ cancers are selected from triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), gastroesophageal cancer, and urothelial carcinoma.

[0279] In some aspects, PTK7+ cancer is a hematological malignancy.

[0280] In some embodiments, PTK7+ cancer is a solid tumor. In some embodiments, PTK7+ cancer is triple-negative breast cancer (TNBC). In some embodiments, PTK7+ cancer is non-small cell lung cancer (NSCLC). In some embodiments, PTK7+ cancer is ovarian malformation cancer. In some embodiments, PTK7+ cancer is pharyngeal cancer. In some embodiments, PTK7+ cancer is gastric adenocarcinoma. In some embodiments, PTK7+ cancer is endometrial adenocarcinoma. In some embodiments, PTK7+ cancer is bladder transitional cell carcinoma. In some embodiments, PTK7+ cancer is bladder transitional cell papilloma. In some embodiments, PTK7+ cancer is esophageal sclerocystic carcinoma (ESCC).

[0281] The methods described herein are intended to reduce tumor size or tumor burden in a subject and / or reduce metastasis in a subject. In various embodiments, the tumor size in a subject is reduced by approximately 25–50%, 40–70%, or 50–90% or more. In various embodiments, the methods reduce tumor size by 10%, 20%, 30%, or more. In various embodiments, the methods reduce tumor size by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. The methods described herein may delay tumor growth.

[0282] As used herein, “subject” refers to a human or an animal. Typically, animals are vertebrates such as primates, rodents, domesticated animals, or game animals. Primates include chimpanzees, crab-eating macaques, spider monkeys, and macaques, e.g., rhesus macaques. Rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Domesticated and game animals include cattle, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cats, canid species, e.g., dogs, foxes, wolves, bird species, e.g., chickens, emus, ostriches, fish, e.g., trout, catfish, and salmon. In certain embodiments, the subject is a mammal, e.g., a primate, e.g., a human. The terms “patient,” “individual,” and “subject” are used interchangeably herein.

[0283] Preferably, the subject is a mammal. The mammal may be, but is not limited to, humans, non-human primates, mice, rats, dogs, cats, horses, or cattle. Non-human mammals can be advantageously used, for example, as representative subjects for various animal models of cancer. Furthermore, the methods described herein can be used to treat livestock and / or pets. The subject may be male or female. In certain embodiments, the subject is human.

[0284] In some embodiments, a subject is identified as having been previously diagnosed with cancer or having cancer and requiring treatment, but does not need to have already received treatment for cancer. In some embodiments, a subject may also not have been previously diagnosed with cancer requiring treatment. In some embodiments, a subject may exhibit one or more risk factors for the condition or one or more cancer-related complications, or may not exhibit any risk factors. In particular, a subject “requiring treatment” for cancer may be a subject who has the condition or has been diagnosed with the condition. In another embodiment, a subject “at risk of developing” the condition refers to a subject who has been diagnosed as being at risk of developing the condition or at risk of re-developing the condition.

[0285] As used herein, the terms “to treat,” “to treat,” “to treat,” or “to improve” as used in relation to a disease, disorder, or medical condition mean a therapeutic action to the condition, the objective of which is to reverse, alleviate, improve, inhibit, slow, or halt the progression or severity of the symptoms or condition. The term “to treat” includes reducing or alleviating at least one side effect or symptom of the condition. Treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective” if the progression of the condition is inhibited or halted. That is, “treatment” includes not only improvement of symptoms or markers but also cessation or at least delay of the progression or worsening of symptoms that would be expected in the absence of treatment. Beneficial or desired clinical outcomes include, but are not limited to, a reduction in cancer cells in the subject, alleviation of one or more symptoms, reduction in the degree of deficit, stabilization of the cancer or malignant tumor condition (i.e., no worsening), delay or slowing of tumor growth and / or metastasis, and extension of life compared to the life expected in the absence of treatment. As used herein, the term “administer” means providing the complex described herein to a subject by a method or route that results in the binding of the complex to cancer cells or malignant cells. Similarly, a pharmaceutical composition containing the complex described herein may be administered by any suitable route that results in an effective therapeutic effect in a subject.

[0286] The dosage range of the complex varies depending on its potency and includes an amount sufficient to produce the desired effect, such as delayed tumor growth or reduction in tumor size. The dosage should not be so high that it causes unacceptable side effects. Generally, the dosage varies depending on the age, condition, and sex of the subject and can be determined by those skilled in the art. The dosage can also be adjusted by the individual physician if any complications occur. In some embodiments, the dosage range is 0.1 mg / kg body weight to 10 mg / kg body weight. In some embodiments, the dosage range is 0.5 mg / kg body weight to 15 mg / kg body weight. In some embodiments, the dosage range is 0.5 mg / kg body weight to 5 mg / kg body weight. Alternatively, the dosage range can be titrated to maintain a blood concentration between 1 ug / mL and 1000 ug / mL. For systemic administration, the target may be given a therapeutic dose, such as 0.1 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 2.5 mg / kg, 5 mg / kg, 10 mg / kg, 12 mg / kg, or more.

[0287] The above doses can be repeated. In a preferred embodiment, the above doses are administered weekly, bi-weekly, every three weeks, or monthly over several weeks or months. The duration of treatment varies depending on the patient's clinical course and response to treatment.

[0288] In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 100 mg / kg. In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 25 mg / kg. In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 20 mg / kg. In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 15 mg / kg. In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 12 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 100 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 25 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 20 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 15 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 12 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 10 mg / kg.

[0289] In some embodiments, the dose may be administered intravenously. In some embodiments, the intravenous administration may be performed by infusion over a period of approximately 10 minutes to approximately 4 hours. In some embodiments, the intravenous administration may be performed by infusion over a period of approximately 30 minutes to approximately 90 minutes.

[0290] In some embodiments, the dose may be administered weekly. In some embodiments, the dose may be administered every other week. In some embodiments, the dose may be administered approximately every two weeks. In some embodiments, the dose may be administered approximately every three weeks. In some embodiments, the dose may be administered every four weeks.

[0291] In some embodiments, the subject receives a total of approximately 2 to 10 doses. In some embodiments, the subject receives a total of 4 doses. In some embodiments, the subject receives a total of 5 doses. In some embodiments, the subject receives a total of 6 doses. In some embodiments, the subject receives a total of 7 doses. In some embodiments, the subject receives a total of 8 doses. In some embodiments, the subject receives a total of 9 doses. In some embodiments, the subject receives a total of 10 doses. In some embodiments, the subject receives a total of more than 10 doses.

[0292] Pharmaceutical compositions containing the complex can be administered in unit doses. When used in reference to pharmaceutical compositions, the term "unit dose" refers to a physically distinct unit suitable as a single dose to a subject, each unit comprising a predetermined amount of the active substance (e.g., the complex) calculated to produce the desired therapeutic effect in conjunction with the necessary physiologically acceptable diluent, i.e., a carrier or vehicle.

[0293] In some embodiments, the complex described herein can be used in a method that includes the step of administering the complex to a subject in need, such as a subject with an autoimmune disease.

[0294] In some embodiments, a method is provided for treating an autoimmune disease, comprising the step of administering a complex described herein. In some embodiments, a subject is in need of treatment for an autoimmune disease. The method described herein comprises the step of administering a therapeutically effective dose of the complex to a subject having an autoimmune disease. As used in this specification, the terms “therapeutic dose,” “effective dose,” or “effective amount” refer to the amount of the complex described herein that provides a therapeutic benefit in the treatment, management, or prevention of relapse of an autoimmune disease, for example, an amount that provides a statistically significant reduction in at least one symptom, sign, or marker of the autoimmune disease. Determining the therapeutic dose is well within the capabilities of those skilled in the art. Generally, the therapeutic dose may vary depending on the subject’s medical history, age, condition, sex, severity and type of the disease in the subject, and the administration of other pharmaceutically active agents.

[0295] The term "autoimmune disease" refers to an immune disorder characterized by the improper activation of immune cells (e.g., lymphocytes or dendritic cells) that impairs the normal functioning of organs and systems of the body. Examples of autoimmune diseases include rheumatoid arthritis, psoriatic arthritis, autoimmune demyelinating diseases (e.g., multiple sclerosis, allergic encephalomyelitis), endocrine ophthalmopathy, uveoretinitis, systemic lupus erythematosus, myasthenia gravis, Graves' disease, glomerulonephritis, autoimmune liver disease, inflammatory bowel disease (e.g., Crohn's disease), anaphylaxis, allergic reactions, Sjögren's syndrome, type 1 diabetes, primary biliary cirrhosis, Wegener's granulomatosis, fibromyalgia, polymyositis, dermatomyositis, polyglandular endocrine insufficiency, Schmidt syndrome, autoimmune uveitis, and Dison's disease, adrenal nephritis, thyroiditis, Hashimoto's disease, autoimmune thyroid disease, pernicious anemia, gastric atrophy, chronic hepatitis, lupus hepatitis, atherosclerosis, subacute cutaneous lupus erythematosus, hypoparathyroidism, Dreysler syndrome, autoimmune thrombocytopenia, idiopathic thrombocytopenic purpura, hemolytic anemia, pemphigus vulgaris, pemphigus, herpetiform dermatitis, alopecia areata, bullous pemphigoid, scleroderma, progressive systemic sclerosis, CREST syndrome (calcification, Raynaud's disease, esophageal motility disorders, kyphosis, and telangiectasia), autoimmune infertility in men and women, ankylosing spine Spondylitis, ulcerative colitis, mixed connective tissue disease, polyarteritis nodosa, systemic necrotizing vasculitis, atopic dermatitis, atopic rhinitis, Goodpasture syndrome, Chagas disease, sarcoidosis, rheumatic fever, asthma, recurrent miscarriage, antiphospholipid antibody syndrome, farmer's lung, erythema multiforme, post-cardiac surgery syndrome, Cushing's syndrome, autoimmune chronic active hepatitis, bird feeder's disease, toxic epidermal necrolysis, Alport syndrome, alveolitis, allergic alveolitis, fibrous alveolitis, interstitial lung disease, erythema nodosum, pyoderma gangrenosum, transfusion reaction, Takayasu's arteritis, rheumatic polysomnitis Myalgia, temporal arteritis, schistosomiasis, giant cell arteritis, ascariasis, aspergillosis, Sumter syndrome, eczema, lymphomatous granulomatosis, Behçet's disease, Kaplan syndrome, Kawasaki disease, dengue fever, encephalomyelitis, endocarditis, endocardial fibrosis, endophthalmitis, persistent erythema elevata, psoriasis, erythroblastosis fetus, eosinophilic fasciitis, Schulman syndrome, Felty syndrome, filariasis, cyclitis, chronic cyclitis, metachronous cyclitis, Fuchs cyclitis, IgA nephropathy, Henoch-Schönlein purpura, graft-versus-host disease, transplant rejection, cardiomyopathy,This includes, but is not limited to, Eaton-Lambert syndrome, relapsing polychondritis, cryoglobulinemia, Waldenström-macroglobulinemia, Evans syndrome, and autoimmune gonadal dysfunction.

[0296] In some embodiments, the methods described herein encompass the treatment of B lymphocyte diseases (e.g., systemic lupus erythematosus, Goodpasture syndrome, rheumatoid arthritis, and type 1 diabetes), Th1 lymphocyte diseases (e.g., rheumatoid arthritis, multiple sclerosis, psoriasis, Sjögren's syndrome, Hashimoto's disease, Graves' disease, primary biliary cholangitis, Wegener's granulomatosis, tuberculosis, or graft-versus-host disease), or Th2 lymphocyte diseases (e.g., atopic dermatitis, systemic lupus erythematosus, atopic asthma, rhinoconjunctivitis, allergic rhinitis, Omen syndrome, systemic scleroderma, or chronic graft-versus-host disease). Generally, diseases involving dendritic cells are associated with Th1 lymphocyte or Th2 lymphocyte diseases.

[0297] As used herein, “subject” refers to a human or an animal. Typically, animals are vertebrates such as primates, rodents, domesticated animals, or game animals. Primates include chimpanzees, crab-eating macaques, spider monkeys, and macaques, e.g., rhesus macaques. Rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Domesticated and game animals include cattle, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cats, canid species, e.g., dogs, foxes, wolves, bird species, e.g., chickens, emus, ostriches, fish, e.g., trout, catfish, and salmon. In certain embodiments, the subject is a mammal, e.g., a primate, e.g., a human. The terms “patient,” “individual,” and “subject” are used interchangeably herein.

[0298] Preferably, the subject is a mammal. The mammal may be, but is not limited to, humans, non-human primates, mice, rats, dogs, cats, horses, or cattle. Non-human mammals can be advantageously used, for example, as representative subjects for animal models of various autoimmune diseases. Furthermore, the methods described herein can be used to treat livestock and / or pets. The subject may be male or female. In certain embodiments, the subject is human.

[0299] In some embodiments, a subject is identified as having or having previously been diagnosed with an autoimmune disease and requiring treatment, but does not need to have already received treatment for an autoimmune disease. In some embodiments, a subject may also not have previously been diagnosed with an autoimmune disease requiring treatment. In some embodiments, a subject may exhibit one or more risk factors for the condition or one or more complications associated with the autoimmune disease, or may not exhibit any risk factors. In particular, a subject “requiring treatment” for an autoimmune disease may be a subject who has the condition or has been diagnosed with the condition. In another embodiment, a subject “at risk of developing” the condition refers to a subject who has been diagnosed as being at risk of developing the condition or at risk of re-developing the condition (e.g., an autoimmune disease).

[0300] As used herein, the terms “to treat,” “to cure,” “to treat,” or “to improve” as used in relation to a disease, disorder, or medical condition mean a therapeutic action to the condition, the objective of which is to reverse, alleviate, improve, inhibit, slow, or halt the progression or severity of the symptoms or condition. The term “to treat” includes reducing or alleviating at least one side effect or symptom of the condition. Treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective” if the progression of the condition is inhibited or halted. That is, “treatment” includes not only improvement of symptoms or markers but also cessation or at least delay of the progression or worsening of symptoms that would be expected in the absence of treatment. Beneficial or desired clinical outcomes include, but are not limited to, a reduction in autoimmune cells in the subject, alleviation of one or more symptoms, reduction in the degree of deficiency, stabilization of the autoimmune disease condition (i.e., no worsening), delay or slowing of the progression of the autoimmune disease, and extension of life compared to the life expected in the absence of treatment. As used herein, the term “administer” means providing the complex described herein to a subject by a method or route that results in the binding of the complex to target autoimmune cells. Similarly, a pharmaceutical composition containing the complex described herein may be administered by any suitable route that results in an effective therapeutic effect in a subject.

[0301] The dosage range of the complex varies depending on its potency and includes an amount sufficient to produce the desired effect, such as delaying the progression of an autoimmune disease or reducing its symptoms. The dosage should not be so high as to cause unacceptable side effects. Generally, the dosage varies depending on the age, condition, and sex of the subject and can be determined by those skilled in the art. The dosage can also be adjusted by the individual physician if any complications arise. In some embodiments, the dosage range is 0.1 mg / kg body weight to 10 mg / kg body weight. In some embodiments, the dosage range is 0.5 mg / kg body weight to 15 mg / kg body weight. In some embodiments, the dosage range is 0.5 mg / kg body weight to 5 mg / kg body weight. Alternatively, the dosage range can be titrated to maintain a blood concentration between 1 ug / mL and 1000 ug / mL. For systemic administration, the target may be given a therapeutic dose, such as 0.1 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 2.5 mg / kg, 5 mg / kg, 10 mg / kg, 12 mg / kg, or more.

[0302] The above doses can be repeated. In a preferred embodiment, the above doses are administered weekly, bi-weekly, every three weeks, or monthly over several weeks or months. The duration of treatment varies depending on the patient's clinical course and response to treatment.

[0303] In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 100 mg / kg. In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 25 mg / kg. In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 20 mg / kg. In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 15 mg / kg. In some embodiments, the dose may be approximately 0.1 mg / kg to approximately 12 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 100 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 25 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 20 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 15 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 12 mg / kg. In some embodiments, the dose may be approximately 1 mg / kg to approximately 10 mg / kg.

[0304] In some embodiments, the dose may be administered intravenously. In some embodiments, the intravenous administration may be performed by infusion over a period of approximately 10 minutes to approximately 4 hours. In some embodiments, the intravenous administration may be performed by infusion over a period of approximately 30 minutes to approximately 90 minutes.

[0305] In some embodiments, the dose may be administered weekly. In some embodiments, the dose may be administered every other week. In some embodiments, the dose may be administered approximately every two weeks. In some embodiments, the dose may be administered approximately every three weeks. In some embodiments, the dose may be administered every four weeks.

[0306] In some embodiments, the subject receives a total of approximately 2 to 10 doses. In some embodiments, the subject receives a total of 4 doses. In some embodiments, the subject receives a total of 5 doses. In some embodiments, the subject receives a total of 6 doses. In some embodiments, the subject receives a total of 7 doses. In some embodiments, the subject receives a total of 8 doses. In some embodiments, the subject receives a total of 9 doses. In some embodiments, the subject receives a total of 10 doses. In some embodiments, the subject receives a total of more than 10 doses.

[0307] The pharmaceutical composition containing the complex can be administered in unit doses. When used in reference to a pharmaceutical composition, the term "unit dose" refers to a physically distinct unit suitable as a single dose to a subject, each unit comprising a predetermined amount of the active substance (e.g., the complex) calculated to produce the desired therapeutic effect in conjunction with the necessary physiologically acceptable diluent, i.e., a carrier or vehicle.

[0308] In some embodiments, the complex, or any of these pharmaceutical compositions, is administered in conjunction with immunosuppressive therapy. In some embodiments, a method is provided for improving treatment outcomes in subjects receiving immunosuppressive therapy. The method generally comprises the steps of administering an effective dose of immunosuppressive therapy to a subject having an autoimmune disease; and administering a therapeutically effective dose of the complex or its pharmaceutical composition that specifically binds to target autoimmune cells to the subject; thereby improving the subject's treatment outcomes compared to administering immunotherapy alone. In some embodiments, the complex is one described herein. In some embodiments, the improved treatment outcomes include reduced disease progression, alleviation of one or more symptoms, etc.

[0309] The description of the aspects of this disclosure is not intended to be exhaustive or to limit the disclosure to the exact form disclosed. Specific aspects and examples of the disclosure are described herein for illustrative purposes, but various equivalent modifications are possible within the scope of the disclosure, as will be recognized by those skilled in the art. The teachings of the disclosure provided herein may be applied to other procedures or methods where appropriate. Further aspects may be provided by combining the various aspects described herein. The aspects of the disclosure may be modified as necessary to provide even further aspects of the disclosure using the above-mentioned references and the structure, function and concepts of the application. These and other modifications may be made to the disclosure in light of the detailed description.

[0310] Certain elements of any of the embodiments described above can be combined or substituted with elements of other embodiments. Furthermore, while advantages related to specific embodiments of the disclosure are described in the context of those embodiments, other embodiments may also exhibit similar advantages, and not all embodiments necessarily exhibit such advantages in order to be included in the scope of the disclosure. All identified patents and other publications are expressly incorporated herein by reference for the purpose of explaining and disclosing methodologies described in such publications that may be used in connection with the present invention, for example. These publications are provided only for their disclosures prior to the filing date of this application. In this regard, nothing should be construed as an acknowledgment that the inventors do not have prior rights to such disclosures due to prior invention or other reasons. All statements regarding the dates or contents of these documents are based on information available to the applicant and do not constitute any acknowledgment of the accuracy of the dates or contents of these documents. [Examples]

[0311] abbreviation Boc2O: Di-tert-butyl bicarbonate Bu4NBr: Tetrabutylammonium bromide DCM: Dichloromethane DEA: Diethanolamine DEAD: Diethyl azodicarboxylate DIPEA: N,N-diisopropylethylamine DMAP: 4-(dimethylamino)pyridine DMF: N,N-dimethylformamide DMTMM: 4-(4,6-dimethoxy-1,3,5-triazine-2-yl)-4-methylmorpholinium chloride DMSO: Dimethyl sulfoxide EEDQ:N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline ESI: Electrospray Ionization HATU: Hexafluorophosphate 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide) HOBt: Hydroxybenzotriazole LCMS: Liquid Chromatography-Mass Spectrometry MeCN: Acetonitrile MeOH: methanol m-CPBA: Metachloroperoxybenzoic acid MTBE: Methyl tert-butyl ether NMR: Nuclear magnetic resonance method Ph3CCl: Triphenylmethyl chloride PNPC: Bis(4-nitrophenyl) carbonate PPh3: Triphenylphosphine TFA: Trifluoroacetic acid THF: Tetrahydrofuran TLC: Thin-layer chromatography TsOH: p-toluenesulfonic acid

[0312] General method 11H NMR and other NMR spectra were recorded using a Bruker AVIII 400 or Bruker AVIII 500. The data were processed with Nuts or MestReNova software to measure the proton shift from the internal standard tetramethylsilane to the low field in parts per million (ppm).

[0313] HPLC-MS measurements were performed on an Agilent 1200 HPLC / 6100 SQ system under the following conditions. Method A: Mobile phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient phase: B increased from 5% to 95% over 15 minutes; Flow rate: 1.0 mL / min; Column: XBridge C18, 4.6*150 mm, 3.5 μm; Column temperature: 40 °C; Detector: ADC ELSD, DAD (214 nm and 254 nm), ES-API. Method B: Mobile phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient phase: B increased from 5% to 95% over 15 minutes; Flow rate: 1.0 mL / min; Column: SunFire C18, 4.6*150 mm, 3.5 μm; Column temperature: 45°C; Detector: ADC ELSD, DAD (214 nm and 254 nm), ES-API. Method C: Mobile phase: A: Water (10 mM NH4HCO3) B: Acetonitrile; Gradient phase: B from 5% to 95% over 15 minutes; Flow rate: 1.0 mL / min; Column: XBridge C18, 4.6*150 mm, 3.5 μm; Column temperature: 40°C. Detector: ADC ELSD, DAD (214 nm and 254 nm), MSD (ES-API).

[0314] LCMS measurements were performed on an Agilent 1200 HPLC / 6100 SQ system under the following conditions. Method A: Mobile phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient phase: B increased from 5% to 95% over 3 minutes; Flow rate: 1.8~2.3 mL / min; Column: SunFire C18, 4.6*50 mm, 3.5 μm; Column temperature: 50°C. Detector: ADC ELSD, DAD (214 nm and 254 nm), ES-API. Method B: Mobile phase: A: Water (10 mM NH4HCO3) B: Acetonitrile; Gradient phase: B from 5% to 95% over 3 minutes; Flow rate: 1.8~2.3 mL / min; Column: XBridge C18, 4.6*50 mm, 3.5 μm; Column temperature: 50°C. Detector: ADC ELSD, DAD (214 nm and 254 nm), MSD (ES-API).

[0315] Preparative high-pressure liquid chromatography (preparative HPLC) was performed on a Gilson 281 under the following conditions. Method A: Waters SunFire 10 μm C18 column (100 Å, 250 × 19 mm). Solvent A was water / 0.01% trifluoroacetic acid (TFA), and solvent B was acetonitrile. The elution condition was a gradient with solvent B linearly increasing from 5% to 100% over 20 minutes at a flow rate of 30 mL / min. Method B: Waters SunFire 10 μm C18 column (100 Å, 250 × 19 mm). Solvent A was water / 0.05% formic acid (FA), and solvent B was acetonitrile. The elution condition was a gradient with solvent B linearly increasing from 5% to 100% over 20 minutes at a flow rate of 30 mL / min. Method C: Waters Xbridge 10 μm C18 column (100 Å, 250 × 19 mm). Solvent A was water / 10 mM ammonium bicarbonate (NH4HCO3), and solvent B was acetonitrile. The elution condition was a gradient with solvent B linearly increasing from 5% to 100% over 20 minutes at a flow rate of 30 mL / min.

[0316] Flash chromatography was performed on Biotage instruments using Agela Flash Column silica-CS; reverse-phase flash chromatography was performed on Biotage instruments using Boston ODS or Agela C18.

[0317] Example 1: Preparation of drug-linker 1 TIFF2026534278000196.tif97146 Process 1 To a solution of 322-8 (600 mg, 1.554 mmol) in DMF (12 mL), DIPEA (602.4 mg, 4.661 mmol) was added, followed by 4,4'-dinitrodiphenyl carbonate (1.42 g, 4.661 mmol). The resulting mixture was then stirred at room temperature for 8 hours until LC-MS detected that 322-8 had been consumed. The reaction mixture was used directly in the next step without workup.

[0318] Process 2 TIFF2026534278000198.tif28128 To the above reaction mixture, HOBt (210 mg, 1.554 mmol), DIPEA (401.7 mg, 3.108 mmol), and 328-6 (746.5 mg, 4.662 mmol) were added sequentially, and the resulting mixture was stirred at room temperature for 6 hours until LC-MS detected that 525-1 had been consumed. The reaction mixture was diluted with ethyl acetate (180 mL), washed with saturated NaHCO3 (aqueous solution, 45 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The crude product was purified by column chromatography (silica, petroleum ether with 0-80% ethyl acetate) to obtain 525-2 (893 mg, 1.56 mmol, 100.4% in two steps) as a pale yellow oil.

[0319] Process 3 A solution of 525-2 (890 mg, 1.555 mmol) in DCM (10 mL) was mixed dropwise with a solution of m-CPBA (483 mg, 2.80 mmol) in DCM (10 mL) at room temperature. The resulting mixture was stirred at this temperature for 24 hours until the 525-2 was consumed, and the reaction was stopped with saturated Na2S2O3 (aqueous solution, 10 mL) and NaHCO3 (aqueous solution, 10 mL). The reaction mixture was stirred for 30 minutes and then diluted with DCM (50 mL). The organic phase was washed with a (aqueous, 1:1, V / V) mixture of saturated Na2S2O3 and NaHCO3 (20 mL x 3), dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure to obtain the crude product. This was purified by column chromatography (silica, petroleum ether with 0-100% ethyl acetate) to obtain 525-3 (790 mg, 1.343 mmol, 86.4%) as a pale yellow oil. Purity = 90-95%.

[0320] Process 4 TIFF2026534278000200.tif20128 A solution of 525-3 (790 mg, 1.343 mmol) in isopropyl alcohol (110 mL) was mixed dropwise with ammonia (110 mL) at room temperature, and the resulting mixture was stirred at this temperature for 12 hours until the 525-3 was consumed. The reaction mixture was concentrated to dryness under reduced pressure to obtain 525-4 (801.2 mg, 1.323 mmol, 98.6%) as a yellow oil, which was used in the next step without further purification. Purity = 90%~95%.

[0321] Process 5 A mixture of 525-4 (801.2 mg, 1.324 mmol), D-glucose (1.43 g, 7.937 mmol), and NaCNBH3 (499.2 mg, 7.94 mmol) in anhydrous MeOH (21 mL) was stirred at 70°C for 24 hours until most of the 525-4 was consumed and 525-5 was detected by LC-MS. The reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure to obtain the crude product, which was purified by reverse-phase liquid chromatography to obtain 525-5 (1.2 g, 1.29 mmol, 97.1%) as a colorless oil with a purity of 85% to 90%.

[0322] Process 6 TIFF2026534278000202.tif37146A mixture of 525-5 (1.2 g, 1.286 mmol) in HCl / MeOH (4 M, 25 mL), MeOH (25 mL), Pd(OH)2 / C (10%, 250 mg), and Pd / C (10%, 250 mg) was stirred at room temperature for 24 hours under a hydrogen atmosphere (balloon) until 525-5 was completely converted to 525-6. The reaction mixture was filtered through a Celite pad, and the filtrate was concentrated to dryness under reduced pressure to obtain 525-6 (886 mg, 1.285 mmol, 100.0%) as an off-white solid, which was used in the next step without further purification.

[0323] Process 7 A solution of 525-7 (104.9 mg, 0.223 mmol) and HATU (305.4 mg, 0.803 mmol) in anhydrous DMF (3 mL) was stirred at room temperature for 15 minutes, then stirred in an ice bath. A solution of 525-6 (600 mg, 0.870 mmol) in anhydrous DMF (3 mL) was followed by dropwise addition of DIPEA (225 mg, 1.74 mmol). The resulting mixture was stirred in an ice bath for 1 hour until most of the 526-7 was consumed. The reaction mixture was purified by reverse-phase liquid chromatography to obtain 525-8 (269.8 mg, 0.113 mmol, 50.9%) as a white solid. Purity = 90%~95%. TIFF2026534278000204.tif24149

[0324] Process 8 To a solution of 525-8 (100 mg, 0.0421 mmol) in MeOH (3 mL) and H2O (1 mL), LiOH·H2O (10.6 mg, 0.252 mmol) was added, and the mixture was stirred at room temperature for 2 hours until the 525-8 was consumed. The reaction mixture was neutralized with 1N HCl to pH=7, concentrated under reduced pressure to obtain the crude product, which was dissolved in H2O (15 mL) and washed with hexane (10 mL x 3). The aqueous phase was concentrated to dryness under reduced pressure to obtain 525-9 (74.6 mg, 0.0346 mmol, 82.3%) as a colorless oil, which was used in the next step without purification.

[0325] Process 9 A solution of 525-9 (70.0 mg, 0.0325 mmol), compound A (49 mg, 0.0360 mmol), HATU (15.1 mg, 0.0397 mmol), and DIPEA (14.0 mg, 0.108 mmol) in anhydrous DMF (4 mL) was stirred at room temperature for 1 hour until compound A was consumed. The reaction mixture was then purified by preparative HPLC to obtain drug-linker 1 (23.5 mg, 0.00672 mmol, 20.7%) as a white solid. LCMS, m / z = 1749.66 (M / 2+H) + m / z = 1166.72 (M / 2+H) + . TIFF2026534278000207.tif38150

[0326] Example 2: Preparation of drug-linker 2 TIFF2026534278000208.tif51128 Process 1 A solution of 330-1 (217.6 mg, 1.48 mmol) and PPh3 (465.5 mg, 1.776 mmol) in THF (8 mL) was prepared, to which a solution of 328-4 (1.3 g, 1.48 mmol) in THF (4 mL) was added, and the mixture was stirred in an ice bath. A solution of DEAD (309.3 mg, 1.776 mmol) in THF (1 mL) was added to the above solution, and the resulting mixture was warmed to room temperature and stirred for 2 hours until 328-4 was consumed by TLC. The reaction was stopped with water (1 mL), and the reaction was concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (silica, petroleum ether with 0-60% ethyl acetate) to obtain 330-2 (1.307 g, 1.297 mmol, 87.7%) as a white solid. Purity=90%~95%. TIFF2026534278000210.tif31150

[0327] Process 2 To a solution of 330-2 (1.472 g, 1.46 mmol) in MeOH (10 mL), N2H4·H2O (146.3 mg, 2.92 mmol) was added in an ice bath. The resulting mixture was warmed to room temperature and stirred for 10 hours until 330-2 was consumed by TLC. The reaction mixture was concentrated under reduced pressure to dryness to obtain the crude product, which was dissolved in ethyl acetate (20 mL) and filtered. The filtrate was concentrated under reduced pressure to obtain 330-3 (1.23 g, 1.402 mmol, 95.9%) as a colorless oil, which was used in the next step without purification.

[0328] Process 3 To a solution of 330-3 (1.23 g, 1.40 mmol) in 8 mL of DCM, Boc2O (367 mg, 1.68 mmol) was added at room temperature. The reaction mixture was stirred at this temperature for 2 hours until 330-3 was consumed and 330-4 was detected by LC-MS. The reaction was concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (silica, petroleum ether with 0-40% ethyl acetate) to obtain 330-4 (1.23 g, 1.26 mmol, 89.8%) as a colorless oil. Purity = 90%-95%.

[0329] Process 4 A solution of 330-4 (800 mg, 0.818 mmol) in DCM (5 mL) was added to a solution of m-CPBA (254 mg, 1.473 mmol) in DCM (5 mL) at room temperature, and the mixture was stirred for 24 hours until the 330-4 was consumed by TLC. Saturated Na2S2O3 (5 mL) and saturated NaHCO3 (5 mL) were added to stop the reaction, and the resulting mixture was stirred for 30 minutes and then diluted with DCM (60 mL). The organic phase was successively washed with saturated Na2S2O3 (20 mL) and saturated NaHCO3 (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated to dryness under reduced pressure. The crude product was purified by column chromatography (silica, petroleum ether, 0-60% ethyl acetate) to obtain 330-5 (689 mg, 0.693 mmol, 84.8%) as a colorless oil. Purity = 90%-95%.

[0330] Process 5 TIFF2026534278000214.tif21146 A solution of 330-5 (689 mg, 0.693 mmol) in isopropyl alcohol (56 mL) was mixed dropwise with ammonia (43 mL) at room temperature, and the resulting mixture was stirred at this temperature for 12 hours until the 330-5 was consumed. The reaction mixture was concentrated to dryness under reduced pressure to obtain 330-6 (698.7 mg, 0.691 mmol, 99.7%) as a yellow oil, which was used in the next step without further purification. Purity = 90%~95%.

[0331] Process 6 A mixture of 330-6 (983 mg, 0.973 mmol), D-glucose (1.05 g, 5.836 mmol), and NaCNBH3 (366.7 mg, 5.836 mmol) in anhydrous MeOH (15 mL) was stirred at 70°C for 24 hours until most of the 330-6 was consumed and 330-7 was detected by LC-MS. The reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure to obtain the crude product, which was purified by reverse-phase liquid chromatography to obtain 330-7 (889 mg, 0.664 mmol, 68.2%) as a colorless oil. Purity = 90%~95%.

[0332] Process 7 A mixture of 330-7 (270 mg, 0.202 mmol) in 25 mL of MeOH, Pd(OH)2 / C (10%, 120 mg), and Pd / C (10%, 120 mg) was stirred at room temperature under a hydrogen atmosphere (balloon) for 24 hours until 330-7 was completely converted to 330-8. The reaction mixture was filtered through a Celite pad, and the filtrate was concentrated to dryness under reduced pressure to obtain 330-8 (138.2 mg, 0.156 mmol, 77.0%) as an off-white solid, which was used in the next step without further purification.

[0333] Process 8 A solution of 330-8 (138 mg, 0.155 mmol) in HCl / MeOH (4 M, 3 mL) and MeOH (3 mL) was stirred at room temperature for 12 hours until 330-8 was completely converted to 330-9. The reaction mixture was concentrated to dryness under reduced pressure to obtain 330-9 (128 mg, 0.155 mmol, 100%) as an off-white solid, which was used in the next step without further purification.

[0334] Process 9 A solution of 330-9 (46 mg, 0.0558 mmol), compound A (53 mg, 0.0390 mmol), HATU (21.2 mg, 0.0558 mmol), and DIPEA (21.6 mg, 0.167 mmol) in anhydrous DMF (4 mL) was stirred at room temperature for 1 hour until compound A was consumed. The reaction mixture was then purified by preparative HPLC to obtain drug-linker 2 (28 mg, 0.0131 mmol, 33.7%) as a white solid. LCMS, m / z = 1066.11 (M / 2+H) + . TIFF2026534278000219.tif38151

[0335] Example 3: Preparation of drug-linker 3 TIFF2026534278000220.tif65146 Process 1 A solution of 328-12 (16.5 mg, 0.0465 mmol) and HATU (38.9 mg, 0.102 mmol) in anhydrous DMF (2 mL) was stirred at room temperature for 15 minutes, and then stirred in an ice bath. A solution of 330-9 (92 mg, 0.112 mmol) in anhydrous DMF (2 mL) was followed by dropwise addition of DIPEA (26.5 mg, 0.205 mmol). The resulting mixture was stirred in an ice bath for 1 hour until most of the 328-12 was consumed. The reaction mixture was purified by reverse-phase liquid chromatography to obtain 330-10 (14 mg, 0.00738 mmol, 15.9%) as a colorless oil.

[0336] Process 2 To a solution of 330-10 (14 mg, 0.00738 mmol) in MeOH (2 mL), LiOH·H2O (2 mg, 0.0442 mmol) was added, and the mixture was stirred at room temperature for 2 hours until the 330-10 was consumed. The reaction mixture was neutralized with 1N HCl to pH=7, concentrated under reduced pressure to obtain the crude product, which was dissolved in H2O (5 mL) and washed with hexane (2 mL × 3). The aqueous phase was concentrated to dryness under reduced pressure to obtain 330-11 (12.4 mg, 0.0074 mmol, 100%) as a white solid, which was used in the next step without purification.

[0337] Process 3 A solution of 330-11 (12 mg, 0.00716 mmol), compound A (5.0 mg, 0.00368 mmol), HATU (2.7 mg, 0.0071 mmol), and DIPEA (2.8 mg, 0.0217 mmol) in anhydrous DMF (2 mL) was stirred at room temperature for 1 hour until compound A was consumed. The reaction mixture was then purified by preparative HPLC to obtain drug-linker 3 (5.0 mg, 0.00166 mmol, 45.1%) as a white solid. LCMS, m / z = 1509.00 (M / 2+H) + m / z = 1006.6 (M / 3+H) + .

[0338] Example 4: Preparation of drug-linker 4 TIFF2026534278000224.tif56146 Process 1 To a solution of 1 (2.5 g, 5.701 mmol) in TIFF2026534278000225.tif40141MeOH (20 mL), D-glucose (4.11 g, 22.804 mmol) and NaBH3CN (1.385 mL, 22.804 mmol) were added. The mixture was stirred under reflux for 24 hours to complete the process. The resulting solution was then concentrated to dryness, and the residue was purified by reverse-phase chromatography (C8 column, eluted with 0-45% methanol in water containing 0.01% TFA) to obtain product 2 as a yellow oil. ESI m / z: 767.5 (M+H) + .

[0339] Process 2 To a solution of 2 (3.3 g, 4.303 mmol) of 2 in MeOH (20 mL), Pd / C (10% wt, 330 mg) was added under nitrogen, and an H2 balloon was attached. The reaction system was degassed, hydrogen was added three times, and then the mixture was stirred at room temperature under a hydrogen atmosphere for 3 hours to complete the reaction. The resulting mixture was filtered to remove catalyst solids, the filtrate was concentrated, and then purified by reverse-phase chromatography (C8 column, elution with 0-25% acetonitrile in water containing 0.01% TFA) to obtain product 3 (2.6 g, 3.510 mmol, 81.50%) as a colorless oil. ESI m / z: 371.3 (M / 2+H)+, 741.4 (M+H) + .

[0340] Process 3 TIFF2026534278000227.tif56146 Compound 5 (0.62 g, 1.755 mmol) was added to a solution of compound 5 in DMF (5 mL) followed by HATU (1...

Claims

1. It includes a heavy chain variable (VH) region and a light chain variable (VL) region, The VH region includes complementarity determination regions HCDR1, HCDR2, and HCDR3 located in the heavy chain variable region framework region. The VL region includes LCDR1, LCDR2, and LCDR3 arranged in the light chain variable region framework region, The VH and VL CD-Rs are These are SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, KVS, and SEQ ID NO:22, respectively. These are SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, FAS, and SEQ ID NO:7, respectively, and These are SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, RTS, and SEQ ID NO:14, respectively. Having an amino acid sequence selected from the set of amino acid sequences shown in the group consisting of, binding substance.

2. The VH region and the VL region are a. SEQ ID NO:15 and SEQ ID NO:16, respectively b. SEQ ID NO:1 and SEQ ID NO:2, respectively, and c. SEQ ID NO:8 and SEQ ID NO:9, respectively. The binding substance according to claim 1, having an amino acid sequence selected from a pair of amino acid sequences represented by the group consisting of the following.

3. The VH region and the VL region are a. SEQ ID NO:15 and SEQ ID NO:16, respectively b. SEQ ID NO:1 and SEQ ID NO:2, respectively, and c. SEQ ID NO:8 and SEQ ID NO:9, respectively. Having an amino acid sequence selected from the amino acid sequence pairs shown in the group consisting of, The heavy chain framework region and the light chain framework region may be modified by substitution, deletion, or insertion of 1 to 8 amino acids within the framework region. The binding substance according to claim 1.

4. The binding substance according to any one of the claims, wherein HCDR1, HCDR2, and HCDR3, and LCDR1, LCDR2, and LCDR3 each have the amino acid sequences shown in SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, KVS, and SEQ ID NO:22, respectively.

5. The binding substance according to claim 1, wherein the framework region is a human framework region.

6. A binding substance according to any one of claims 1 to 5, which is an antibody or its antigen-binding portion.

7. The conjugate according to any one of the claims, which is a monoclonal antibody, Fab, Fab', F(ab'), Fv, disulfide-bonded Fc, scFv, single-domain antibody, diabody, bispecific antibody, or multispecific antibody.

8. The binding substance according to any one of the claims, wherein the heavy chain variable region further comprises a heavy chain steady region.

9. The binding substance according to claim 8, wherein the heavy chain constant region is of the IgG isotype.

10. The binding substance according to claim 9, wherein the heavy chain constant region is the IgG1 constant region.

11. The binding substance according to claim 8, wherein the heavy chain constant region is the IgG4 constant region.

12. The binding substance according to claim 10, wherein the IgG1 constant region has the amino acid sequence shown in SEQ ID NO:

56.

13. The binding substance according to any one of the claims, wherein the light chain variable region further comprises a light chain steady region.

14. The binding substance according to claim 13, wherein the light chain constant region is of the κ-isotype.

15. The binding substance according to claim 14, wherein the light chain constant region has the amino acid sequence shown in SEQ ID NO:

57.

16. The binding substance according to any one of claims 8 to 15, wherein the heavy chain constant region further comprises at least an amino acid modification that reduces the binding affinity to human FcγRIII.

17. A binding substance according to any one of the above claims, which is single-specific.

18. A divalent binding substance according to any one of claims 1 to 17.

19. A binding substance according to any one of claims 1 to 16, which is bispecific.

20. A binding substance according to any one of claims 1 to 19, which specifically binds to protein tyrosine kinase 7 (PTK7).

21. The VH region has the amino acid sequence shown in SEQ ID NO:15, The VL region has the amino acid sequence shown in SEQ ID NO:16, The heavy chain constant region has the amino acid sequence shown in SEQ ID NO:56, The light chain constant region has the amino acid sequence shown in SEQ ID NO:

56. The binding substance according to any one of claims 1 to 20.

22. A pharmaceutical composition comprising a binder according to any one of claims 1 to 21 and a pharmaceutically acceptable carrier.

23. A nucleic acid encoding the binding substance according to any one of claims 1 to 21.

24. A vector comprising the nucleic acid described in claim 23.

25. A cell line comprising the vector according to claim 23 or the nucleic acid according to claim 24.

26. A binding substance according to any one of claims 1 to 21, At least one linker bonded to the bonding material, At least one drug unit and Includes, Here, each drug unit is bound to a linker, and the linker may contain at least one polar group. A complex.

27. The linker is derived from a linker compound or its stereoisomer or salt. The linker compound is Linker units and A stretcher unit connected to the linker unit, The amino acid unit is arbitrary, at least one polar group and Including, here, The stretcher group has a binding site to the binding substance and a binding site to the amino acid unit (if present) or linker subunit, The amino acid unit (if present) has a binding site to a stretcher unit and a binding site to a linker unit, The linker unit has binding sites to the amino acid unit (if present) or to the stretcher unit and to the at least one drug unit, The composite according to claim 26.

28. The composite according to claim 27, wherein the linker unit is inseparable.

29. The complex according to claim 27, wherein the polar group is bonded to the amino acid unit or the stretcher group.

30. The aforementioned linker compound (a) Having 1 to 4 binding sites for the drug unit, and having the following structure (i) or (ii): The linker unit having one of the following, (b) At least one polar group each comprising a polymer unit, (c) The stretcher group having a binding site for the binding substance and Includes, During the ceremony, α- is a binding site to a group that can be cleaved by enzymes. β- is the binding site to the at least one polar group, δ- is H, a binding site to at least one of the drug units, or a binding site to a linking group attached to at least one of the drug units. The polymer unit comprises a polyamide, a polyether, or a combination thereof, wherein the polyether comprises a hydroxyl group, a polyhydroxyl group, a sugar group, a carboxyl group, or a combination thereof. Each R a H or C 1 ~C 6 It is alkyl, Each R b They are independent of Haro, C 1～6 The alkyl group is a binding site to at least one of the drug units, or a binding site to at least one of the polar groups. x is 0, 1, 2, 3, or 4. y is 0, 1, 2, or 3. R c is a bond, -C(O)-, -S(O)-, -SO 2 -, C 1～6 alkylene, C 1～6 alkynylene, triazolyl, or a combination thereof, Y is a bond, -O-, -S-, -N(R a )-, -C(O)-, -S(O)-, -SO 2 -C 1 ~C 6 Alkilen, C 1 ~C 6 Alkenylene, C 1 ~C 6 Alkinylene, triazolyl, or a combination thereof. The composite according to claim 27.

31. The aforementioned linker unit has the following structure (ia) or (ii-a): The composite according to claim 30, comprising one of the following or a stereoisomer or salt thereof.

32. The aforementioned linker unit has one of the following structures: (ib), (ic), (id), (ie), or (if): The composite according to claim 30, comprising one of the following or a stereoisomer or salt thereof.

33. The aforementioned at least one polar group is given by the following formula: L3-N(CH 2 -(CH(XR)) k -X 1 (X 2 )) 2 (X) or comprising at least one sugar unit having a stereoisomer or salt thereof, During the ceremony, Each X is independently selected from NH and O. Each R is independently selected from hydrogen, acetyl, monosaccharides, disaccharides, and polysaccharides. each X １ CH is independent 2 And selected from C(O), each X 2 These are independently selected from H, OH, and OR. k is between 1 and 10. L3 is the bonding point to the rest of the polar group. The composite according to any one of claims 30 to 32.

34. The aforementioned at least one polar group has the following structure (XII) or (XIII): It comprises at least one sugar unit having one of the following or a stereoisomer or salt thereof, During the ceremony, Each R is independently selected from hydrogen, monosaccharides, disaccharides, and polysaccharides. m is between 1 and 8. n is between 0 and 4. The composite according to any one of claims 30 to 32.

35. The following formula: (a) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -NR 24 R 25 (XX) or its stereoisomer, salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is alkylene, R 24 and R 25 These are independently H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, and substituted C 1 ~C 8 Alkyl, substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More will be selected, however, R 24 and R 25 It is not possible for both to be H. n20 is 2 to 26; or (b) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -NR 24 R 25 (XXI) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is alkylene, R 24 and R 25 One of them consists of H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and a substituted C 1 ~C 8 Alkyl, substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More selected, R 24 and R 25 The other is polyethylene glycol which may have 1 to 24 ethylene glycol subunits. n20 is 2 to 26; or (c) ~R 20 -[-R 26 -[R 29 -[O-CH 2 -CH 2 -] n20 R 29 ] n21 -R 27 -NR 24 R 25 ] n27 (XXII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 26 and R 27 are each independently optional, and are independently selected from a bond, C 1 to C 12 alkylene, -NH-C 1 to C 12 alkylene, -C 1 to C 12 alkylene-NH-, -C 1 to C 12 alkylene-N(CH 3 )-, -C(O)-C 1 to C 12 alkylene, -C 1 to C 12 alkylene-C(O)-, -NH-C 1 to C 12 alkylene-C(O)- and -C(O)-C 1 to C 12 alkylene-NH- R 24 and R 25 One of them consists of H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and a substituted C 1 ~C 8 Alkyl, substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More selected, R 24 and R 25 The other is selected from polyethylene glycol which may have H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and 1 to 24 ethylene glycol subunits, where R 28 is a sugar unit of formula (XII) or (XIII), however, R 24 and R 25 It is not possible for both to be H. Each R 29 These are arbitrary and can be -C(O)-, -NH-, and -C(O)-C independently. 1 ~C 6 Alkylene-,-NH-C 1 ~C 6 Alkylene-,-C 1 ~C 6 Alkylene-NH-,-C 1 ~C 6 Alkylene-C(O)-,-NH(CO)-C 1 ~C 6 Alkylene-,-N(CH) 3 )-(CO)-C 1 ~C 6 Selected from alkylene-, -NH(CO)NH-, and triazole, n20 ranges from 2 to 26. n21 is 1 to 4. n27 is 1 to 4; or (d) ~R 20 -R 21 -[-C(R α )H-C(O)-N(R N )-] n20 -R 22 -NR 24 R 25 (XXIII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 is a combination or C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 ,-[CH 2 -CH 2 -O] n20 -C 1 ~C 3 Alkylene-, or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 -C(O)-, R 22 is C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 ,-[CH 2 -CH 2 -O] n20 -C 1 ~C 3 Alkylene- or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 -C(O)-, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N H and C are independent of each other. 1 ~C 6 Alkyl or -R 22 -NR 24 R 25 And, R 24 and R 25 These are independently H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, and substituted C 1 ~C 8 Alkyl, substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More will be selected, however, R 24 and R 25 It is not possible for both to be H. Each n20 is independently 2 to 26; or (e) ~R 20 -R 21 -[-C(R α )HC(O)-N(R N )-] n20 -R 22 -WHAT 2 R 26 (XXIV) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each is independent, combined, C 1 ~C 3 Alkylene, or -C 1 ~C 3 Alkylene [O-CH 2 -CH 2 -] n20 And, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N H and C are independent of each other. 1 ~C 6 Alkyl or -R 22 -NR 24 R 25 And, R 24 and R 25 These are independently H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, and substituted C 1 ~C 8 Alkyl, substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More will be selected, however, R 24 and R 25 It is not possible for both of them to be H. R 26 is H or C 1 ~C 4 It is alkyl, Each n20 is independently between 2 and 26, However, at least one R α or R N ha-R 22 -NR 24 R 25 is; or (f) ~R 20 -R 21 -[C(R α )H-C(O)-N(R N )-] n20 -R 22 -N-(R 23 -NR 24 R 25 ) 2 (XXV) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each is independent, combined, C 1 ~C 3 Alkylene, or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 And, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N H or C 1 ~C 6 It is alkyl, Each R 23 C is independent 1 ~C 6 It is alkylene, R 24 and R 25 These are independently H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, and substituted C 1 ~C 8 Alkyl, substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More will be selected, however, R 24 and R 25 It is not possible for both of them to be H. Each n20 is independently between 2 and 26; A composite according to any one of claims 30 to 34, comprising a polar group having the same polar group.

36. R 24 and R 25 Each is independently selected from H and polyhydroxyl groups, but R 24 and R 25 The composite according to claim 35, wherein neither of them can be H.

37. The complex according to claim 35 or 36, wherein the polyhydroxyl group is a linear monosaccharide which may be selected from C6 or C5 sugars, sugar acids, and amino sugars.

38. The C6 or C5 sugar is selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, growth, idose, talose, aldose, and ketose. The aforementioned sugar acid is selected from gluconic acid, aldonic acid, uronic acid, and urosonic acid, or The amino sugar is selected from glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine. The composite according to claim 37.

39. below: or comprising a polar group selected from their stereoisomers or salts, During the ceremony, Each R is independently either H or alkyl. Each R 39 These are independently selected from H, a linear monosaccharide, and polyethylene glycol which may have 1 to 24 ethylene glycol subunits. Each n is independently between 1 and 12. The wavy line indicates either region β or region R. b A binding site to, or to the enzyme-cleavable group, The composite according to any one of claims 35 to 38.

40. The complex according to any one of claims 30 to 39, wherein the binding site β is formed from a functional group of the precursor compound of the polar group, and the functional group is selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctin, hydrazine, carbonylalkyl heteroaryl, and their protected forms.

41. below: (a) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -R 30 (XXX) or its stereoisomer or salt, in the formula, R 20 This is directed to site β, and site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R 30 C may be substituted 3 ~C 10 Selected from carbocyclic, thiourea, optionally substituted thiourea, urea, optionally substituted urea, sulfamide, alkylsulfamide, acylsulfamide, optionally substituted alkylsulfamide, optionally substituted acylsulfamide, sulfonamide, optionally substituted sulfonamide, guanidine containing alkyl and arylguanidine, phosphoramide, or optionally substituted phosphoramide, or R 30 These include azide, alkynyl, substituted alkynyl, -NH-C(O)-alkynyl, and -NH-C(O)-alkynyl-R 65 , cyclooctin, -NH-cyclooctin, -NH-C(O)-cyclooctin, or -NH-(cyclooctin) 2 Selected from, here, R 65 This is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbon ring, optionally substituted aryl, optionally substituted heterocarbon ring, or optionally substituted heteroaryl. n20 is between 2 and 26; (b) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -NH-C(O)-R 31 (XXXI) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R 31 It is a branched polyethylene glycol chain, where each branch has 1 to 26 ethylene glycol subunits and each branch has R at its end. 35 It has, R 35 azid, alkinyl, alkinyl-R 65 , cyclooctin or cyclooctin-R 65 And here, R 65 This is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbon ring, optionally substituted aryl, optionally substituted heterocarbon ring, or optionally substituted heteroaryl. n20 is between 2 and 26; (c) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -C(O)NH-R 31 (XXXII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R 31 This is a branched polyethylene glycol chain, where each branch independently has 1 to 26 ethylene glycol subunits, and each branch has R at its end. 35 It has, R 35 azid, alkinyl, alkinyl-R 65 , cyclooctin or cyclooctin-R 65 And here, R 65 This is selected from optionally substituted alkyls, optionally substituted alkenyls, optionally substituted alkynyls, optionally substituted carbon rings, optionally substituted aryls, optionally substituted heterocarbon rings, and optionally substituted heteroaryls. n20 is between 2 and 26; (d) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -C(O)NR 31 -R 22 -NR 24 R 25 (XXXIII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 31 is H or R 22 -NR 24 R 25 And, R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, and a substituted -C(O)-polyhydroxyl group, however, R 24 and R 25 It is not possible for both of them to be H. n20 is between 2 and 26; (e) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -N(R 33 -R 31 ) 2 (XXXIV) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R 31 It is a branched polyethylene glycol chain, where each branch has 1 to 26 ethylene glycol subunits and each branch has R at its end. 35 It has, R 33 is C 1 ~C 3 Alkilen, C 1 ~C 3 Alkylene-C(O),-C(O)-C 1 ~C 3 Alkylene, or -C(O)-C 1 ~C 3 It is alkylene-C(O), R 35 azid, alkinyl, alkinyl-R 65 , cyclooctin or cyclooctin-R 65 And here, R 65 This is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbon ring, optionally substituted aryl, optionally substituted heterocarbon ring, or optionally substituted heteroaryl. n20 is between 2 and 26; (f) ~R 20 -(R 21 -[CH 2 -CH(OR 34 )-CH 2 -O] n20 -R 36 ) n25 (XXXV) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. Each R 21 These are independent, bonded, -O-, or C 1 ~C 3 It is an alkylene group, Each R 34 These are independently H, -[CH 2 -CH(OH)-CH 2 -O] n20 -R 36 -C(O)-NR 24 R 25 or -C(O)N(R N )-C 1 ~C 6 Alkilen-NR 24 R 25 And, R N is H or C 1 ~C 4 It is alkyl, R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, however, R 24 and R 25 It is not possible for both of them to be H. Each R 36 H and C are independent of each other. 1 ~C 6 Alkylene-C(OH)H-NR 44 R 45 , C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkilen-NR 44 R 45 -C(O)-NR 24 R 25 ,-C(O)N(R N )-C 1 ~C 6 Alkilen-NR 24 R 25 , C 1 ~C 6 Alkylene-C(O)NR 24 R 25 or C 1 ~C 6 Alkylene-CO 2 R 37 And, Each R 37 H or C 1 ~C 6 It is alkyl, R 44 and R 45 Each of these is independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, and a substituted -C(O)-polyhydroxyl group, however, R 44 and R 45 It is not possible for both of them to be H. Each n20 is independently between 1 and 26. n25 is either 1 or 2; (g) ~R 20 -R 21 -[[CH 2 -CH 2 -O] n20 -R 22 -[CH 2 -[CH(OH)] n23 -CH 2 -O] n21 ] n22 -R 23 -NR 24 -R 25 (XXXVI) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 , R 22 and R 23 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, and a substituted -C(O)-polyhydroxyl group, however, R 24 and R 25 It is not possible for both of them to be H. Each n20 is independently between 0 and 26, and each n21 is independently between 0 and 26, provided that at least one of n20 or n21 is between 2 and 26. n22 is 1 to 5. Each n23 is independently either 1 or 2; (h) ~R 20 -(R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -N(R N )-CO 2 -[CH 2 -CH(OR 34 )-CH 2 -O] n21 -R 36 ) n25 (XXXVII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R N is H or C 1 ~C 4 It is alkyl, R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, and a substituted polyhydroxyl group, however, R 24 and R 25 It is not possible for both of them to be H. Each R 34 These are independently H, -[CH 2 -CH(OH)-CH 2 -O] n20 -R 36 , or -C(O)N(R N )-C 1 ~C 6 Alkilen-NR 24 R 25 And, Each R 36 H and C are independent of each other. 1 ~C 6 Alkylene-C(OH)H-NR 44 R 45 , C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkilen-NR 44 R 45 ,-C(O)N(R N )-C 1 ~C 6 Alkilen-NR 24 R 25 , C 1 ~C 6 Alkylene-C(O)NR 24 R 25 or C 1 ~C 6 Alkylene-CO 2 R 37 And, Each R 37 H or C 1 ~C 6 It is alkyl, R 44 and R 45 Each of these is independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, and a substituted -C(O)-polyhydroxyl group, however, R 44 and R 45 It is not possible for both of them to be H. n20 ranges from 2 to 26. n21 ranges from 1 to 26. n25 is either 1 or 2; (i) ~R 20 -(R 21 -[N(R N )-C(O)-[O-CH 2 -CH(OH)-CH 2 ] n20 ] n21 -R 22 -NR 24 R 25 ) n25 (XXXVIII) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R N is H or C 1 ~C 4 It is alkyl, R 24 and R 25 Each of these is independently selected from H, a polyhydroxyl group, and a substituted polyhydroxyl group, however, R 24 and R 25 It is not possible for both of them to be H. n20 ranges from 2 to 26. n21 is 1 to 4. n25 is 1, 2, or 3; (j) ~R 20 -(R 21 -[C(R α )H-C(O)-N(R N )] n20 -R 22 -[CH 2 -CH 2 -O] n20 -NR 24 R 25 ) n25 (XXXIX) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each is independent, combined, C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 ,-[CH 2 -CH 2 -O] n20 -C 1 ~C 3 Alkylene- or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 -C(O)-, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N H and C are independent of each other. 1 ~C 6 Alkyl or -R 22 -NR 24 R 25 And, R 24 and R 25 Each of these is independently H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More will be selected, however, R 24 and R 25 It is not possible for both of them to be H. Each n20 is independently between 0 and 26, however, at least one of the n20s is between 2 and 26. n25 is 1 or 2; or (k) ~R 20 -R 21 -[C(R α )H-C(O)-N(R N )] n20 -R 22 -[CH 2 -CH 2 -O] n20 -NR 24 R 25 R 21 -[C(R α )H-C(O)-N(R N )] n21 -R 22 -[CH 2 -CH 2 -O] n21 -R 23 -CO 2 -R 26 (XXXVX) or its stereoisomer or salt, in the formula, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 , R 22 and R 23 Each is independent, combined, C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 ,-[CH 2 -CH 2 -O] n20 -C 1 ~C 3 Alkylene- or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 -C(O)-, Each R α H or -R are independent. 22 -NR 24 R 25 And, Each R N H and C are independent of each other. 1 ~C 6 Alkyl or -R 22 -NR 24 R 25 And, R 24 and R 25 Each of these is independently H, polyhydroxyl group, substituted polyhydroxyl group, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More will be selected, however, R 24 and R 25 It is not possible for both of them to be H. R 26 is H or C 1 ~C 6 It is alkyl, Each n20 is independently between 0 and 26, except that at least one n20 is between 2 and 26. Each n21 is independently between 0 and 26, except that at least one n21 is between 2 and 26; The composite according to any one of claims 30 to 34, comprising a polar group having a more selectable formula.

42. below: ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -NH-C(O)-R 31 (XXXI)、 ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -C(O)NH-R 31 (XXXII), and ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -N-(R 33 -R 31 ) 2 (XXXIII) A polar group comprising a more selected formula or its stereoisomer or salt, During the ceremony, R 20 is directed to site β or site R b It is a binding group to, or to, a group that can be enzymatically cleaved. R 21 and R 22 Each can be joined or C independently 1 ~C 3 It is an alkylene group, R 31 It is a branched polyethylene glycol chain, where each branch has 1 to 26 ethylene glycol subunits and each branch has R at its end. 35 It has, R 33 is C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-C(O),-C(O)-C 1 ~C 3 Alkylene or -C(O)-C 1 ~C 3 It is alkylene-C(O), R 35 azid, alkinyl, alkinyl-R 65 , cyclooctin or cyclooctin-R 65 And here, R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbon ring, optionally substituted aryl, optionally substituted heterocarbon ring, or optionally substituted heteroaryl, and the wavy (~) line is R 20 This shows the binding site, and n20 is 2 to 26. The composite according to any one of claims 30 to 34.

43. The complex according to any one of claims 41 to 42, wherein the bond to site β, site Rb, or the enzymatically cleavable group is formed from a functional group of the precursor compound of the polar group, the functional group being selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctin, hydrazine, carbonylalkyl heteroaryl, and their protected forms.

44. formula: ~R 20 -(R 43 -R 41 -[O-CH 2 -CH 2 ] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XL) or a polar group having a stereoisomer or salt thereof, During the ceremony, R 20 is directed to site β or site R b A binding group to, or to the enzymatically cleavable group, R 41 and R 42 Each can be joined or C independently 1 ~C 6 It is alkylene, Each R 43 They are independent, combined, C 1 ~C 12 Alkylene, -OC 1 ~C 12 Alkylene, -C(=O)-, -NR a -C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkilen-NR a -, -C(O)-C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkylene-C(O)-,-NR a -C 1 ~C 12 Alkylene-C(O)-,-C(O)-C 1 ~C 12 Alkilen-NR a -, -NR a -C(O)-NR a -, -NR a -C(O)-, -NR a -C(O)-C 1 ~C 12 Alkylene, -C(O)-NR a -C 1 ~C 12 Alkylene, -heteroarylene, heteroaryl-C 1 ~C 12 Alkylene, heteroaryl-C 1 ~C 12 Alkylene-C(O)-, or -C(O)NR 46 R 47 Further selection, where each alkylene is hydroxyl, SO 3 H and / or oxo may be substituted, R a H, C 1 ~C 6 It is an alkyl group, a polyhydroxyl group, or a substituted polyhydroxyl group, R 46 and R 47 One of them is H or C 1 ~C 12 It is alkylene, and the other is C 1 ~C 12 It is alkylene, and here, C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom 44 R 45 It is connected, R 44 and R 45 Each of these is independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituent is selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and n40 is 2 to 26, however, R 44 and R 45 It is not possible for both of them to be H. n40 ranges from 2 to 26. n41 is 1 to 6. n42 is 1 to 6. The composite according to any one of claims 30 to 34.

45. formula: ~R 20 -(R 41 -[O-CH 2 -CH 2 ] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLI) or a polar group having a stereoisomer or salt thereof, During the ceremony, R 20 is directed to site β or site R b A binding group to, or to the enzymatically cleavable group, R 41 and R 42 Each can be joined or C independently 1 ~C 6 It is alkylene, R 43 is a bond, C 1 ~C 12 Alkylene, -OC 1 ~C 12 Alkylene, -C(=O)-, -NR a -C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkilen-N'R a -, -C(O)-C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkylene-C(O)-,-NR a -C 1 ~C 12 Alkylene-C(O)-,-C(O)-C 1 ~C 12 Alkilen-NR a -, -NR a -C(O)-NR a -, -NR a -C(O)-, -NR a -C(O)-C 1 ~C 12 Alkylene, C(O)-NR a -C 1 ~C 12 Alkylene, -heteroarylene, heteroaryl-C 1 ~C 12 Alkylene, heteroaryl-C 1 ~C 12 Alkylene-C(O)- and -C(O)NR 46 R 47 Further selection, where each alkylene is hydroxyl, SO 3 H and / or oxo may be substituted, R a H, C 1 ~C 6 It is an alkyl group, a polyhydroxyl group, or a substituted polyhydroxyl group, R 46 and R 47 One of them is H or C 1 ~C 12 It is alkylene, and the other is C 1 ~C 12 It is alkylene, and here, C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom 44 R 45 It is connected, R 44 and R 45 Each of these is independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituent is selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, provided that R 44 and R 45 It is not possible for both of them to be H. n40 ranges from 1 to 26. n41 is 1 to 6. n42 is 1 to 6. The composite according to any one of claims 30 to 34.

46. formula: ~R 20 -(R 41 -[O-CH 2 -CH 2 ] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLII) or a polar group having a stereoisomer or salt thereof, During the ceremony, R 20 is directed to site β or site R b A binding group to, or to the enzymatically cleavable group, R 41 and R 42 are each independently a bond or C 1 to C 3 alkylene. R 43 is a bond, C 1 ~C 6 Alkylene, -OC 1 ~C 12 Alkylene, -C(=O)-, -NR a -C 1 ~C 12 Alkylene, -C 1 ~C 6 Alkilen-NR a -, -C(O)-C 1 ~C 6 Alkylene, -C 1 ~C 6 Alkylene-C(O)-,-NR a -C 1 ~C 6 Alkylene-C(O)-,-C(O)-C 1 ~C 6 Alkilen-NR a -, -NR a -C(O)-NR a -, -NR a -C(O)-, -NR a -C(O)-C 1 ~C 6 Alkylene, -C(O)-NR a -C 1 ~C 12 Alkylene, -heteroarylene, heteroaryl-C 1 ~C 6 Alkylene, heteroaryl-C 1 ~C 6 Alkylene-C(O)- and -C(O)NR 46 R 47 Further selection, where each alkylene is hydroxyl, SO 3 H and / or oxo may be substituted, R a H, C 1 ~C 6 It is an alkyl group, a polyhydroxyl group, or a substituted polyhydroxyl group, R 46 and R 47 One of them is H or C 1 ~C 6 It is alkylene, and the other is C 1 ~C 12 It is alkylene, and here, C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom 44 R 45 It is connected, R 44 and R 45 Each of these is independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituent is selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, provided that R 44 and R 45 It is not possible for both of them to be H. n40 ranges from 1 to 16. n41 is 1 to 4. n42 is 1 to 4. The composite according to any one of claims 30 to 34.

47. R 20 The complex according to any one of claims 35, 41-42 and 44-46, wherein the functional group is formed from a functional group of the precursor compound of the polar group, and the functional group is selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctin, hydrazine, carbonylalkyl heteroaryl, or a protected form thereof.

48. R 20 However, the structure is as follows: It includes one of the following or its stereoisomer, During the ceremony, R is H, C 1 ~C 6 It is an alkyl or polyhydroxyl group, n is between 0 and 12. is directed to site β or site R b This indicates binding to, or to the enzyme-cleavable group, This indicates the binding site to the remaining portion of the polar group. The composite according to any one of claims 35, 41-42, and 44-46.

49. R 20 However, the structure is as follows: Having one of the following or its stereoisomer, During the ceremony, n is between 0 and 12. is directed to site β or site R b This indicates binding to, or to the enzyme-cleavable group, This indicates the binding site to the remaining portion of the polar group. The compound according to any one of claims 35, 41-42, and 44-46.

50. R 43 -(NR 44 R 45 )n 41 has the following structure: Having one of the following or its stereoisomer, During the ceremony, R a H, C 1～6 Alkyl, polyhydroxyl, or substituted polyhydroxyl groups, p is an integer from 1 to 6. R to the remaining portion of the polar group 43 The binding site is shown. The composite according to any one of claims 44 to 49.

51. R 43 -(NR 44 R 45 )n 41 However, the structure is as follows: Having one of the following or its stereoisomer, During the ceremony, R to the remaining portion of the polar group 43 The binding site is shown. The composite according to any one of claims 44 to 49.

52. -NR 44 R 45 However, the structure is as follows: Having one of the following or its stereoisomer, During the ceremony, -NR to the remaining portion of the polar group 44 R 45 The binding site is shown. The composite according to any one of claims 44 to 51.

53. The following structure before bonding to the aforementioned linker unit: It contains a polar group having one of the following, During the ceremony, (*) indicates site β or site R b The site indicates a binding site to or to the enzyme-cleavable group, Each R is independently H or C 1 ~C 6 It is alkyl, R' is H, C 1 ~C 6 Alkyl, -N(R 24 )(R 25 ) or -CO 2 H is, Each n is independently between 1 and 12. X is O, NR, or -CH 2 -and, V is a combination or C 1 ~C 6 It is alkyl, R 24 and R 25 One of them consists of H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and a substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, and R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 More selected, R 24 and R 25 The other side consists of H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, and a substituted -C(O)-C 1 ~C 8 Alkyl, chelating agent, R 28 -C(O)-R is the sugar unit of formula (XII) or (XIII). 28 , and polyethylene glycol which may have 1 to 24 ethylene glycol subunits, however, R 24 and R 25 It is not possible for both to be H. The composite according to any one of claims 30 to 52.

54. below: or comprising a polar group selected from their stereoisomers or salts, During the ceremony, Each Z is connected at *, independently More selected, each is directed to site β or site R b This is a bond to, or to the enzymatically cleavable group, The composite according to any one of claims 30 to 34 or 44 to 46.

55. A composite according to any one of claims 30 to 54, comprising a polar group containing polymer units and sugar units.

56. The composite according to any one of claims 30 to 54, comprising a polar group containing at least two polymer units.

57. A composite according to any one of claims 30 to 54, comprising a polar group containing polymer units and carboxyl units.

58. A complex according to any one of claims 30 to 54, comprising at least two polar groups.

59. A composite according to any one of claims 30 to 54, comprising a polar group containing polymer units, sugar units, and carboxyl units.

60. The composite according to any one of claims 30 to 54, comprising a polar group having at least two polymer units, at least one sugar unit, and at least one carboxyl unit.

61. The complex according to any one of claims 30 to 54, wherein the enzyme-cleavable group comprises at least two of the amino acid units.

62. The complex according to any one of claims 30 to 61, comprising at least one polar group bound to the enzyme-cleavable group.

63. The following structure: Having one of the following, During the ceremony, Rc is a bond or C1-6 alkylene. The wavy lines on the amino group indicate a binding site for a stretcher group, or indicate H prior to binding to the stretcher group. β- is a binding site to at least one polar group, The benzyl H on the benzyl OH may be replaced by a bond to one of the drug units or to the linking group bonded to at least one of the drug units. The composite according to any one of claims 30 to 62.

64. The complex according to any one of claims 30 to 63, wherein the enzyme-cleavable group comprises a peptide that can be cleaved by an intracellular protease.

65. The complex according to claim 64, wherein the intracellular protease is cathepsin B.

66. The complex according to claim 65, wherein the enzyme-cleavable group comprises a cleavable peptide including valine-citrulline peptide, valine-alanine peptide, valine-lysine peptide, phenylalanine-lysine peptide, or glycine-glycine-phenylalanine-glycine peptide.

67. The following structure: Includes one of the following: During the ceremony, R C is a combination or C 1～6 It is alkylene, The wavy line on the amino group indicates a binding site for the stretcher group, or indicates H prior to binding to the stretcher group. β- is a binding site to at least one polar group, The H on the benzyl OH may be replaced by a bond to one of the drug units, or a bond to the binding site to at least one of the drug units. The composite according to any one of claims 64 to 66.

68. The following structure: Having one of the following, During the ceremony, The wavy line on the amino group indicates the binding site to the stretcher group, or indicates H before binding to the stretcher group. The hydrogen atoms on the benzyl OH group may be replaced by a bond to one of the drug units, or to a linking group bonded to at least one of the drug units. The composite according to any one of claims 30 to 33.

69. The complex according to any one of claims 30 to 68, wherein the enzyme-cleavable group is bonded to the stretcher group by a non-peptide linking group.

70. The non-peptide linking group may be substituted with C 1 ~C 10 Alkylene, C may be substituted. 2 ~C 10 Alkenylene, C may be substituted. 2 ~C 10 The composite according to claim 69, selected from alkylylene or optionally substituted polyethylene glycol.

71. The complex according to any one of claims 30 to 70, comprising the stretcher group bound to the enzyme-cleavable group.

72. The stretcher group is as follows: More selected, During the ceremony, R 17 is -C 1 ~C 10 Alkylene-,-C 1 ~C 10 Heteroalkylene-,-C 3 ~C 8 Carbocyclo-,-O-(C 1 ~C 8 Alkylene)-,-(CH 2 -O-CH 2 ) b -C 1 ~C 8 Alkylene-(where b is 1 to 26), -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -(where b is between 1 and 26), -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C 1 ~C 8 Alkylene-(where b is 1 to 26), -arrine-, -C 1 ~C 10 Alkilen-Arirene-, -Arirene-C 1 ~C 10 Alkylene-,-C 1 ~C 10 Alkylene-(C) 3 ~C 8 Carbocyclo)-,-(C 3 ~C 8 Carbocyclo)-C 1 ~C 10 Alkylene-,-C 3 ~C 8 Heterocyclo-, -C 1 ~C 10 Alkylene-(C) 3 ~C 8 Heterocyclo)-,-(C 3 ~C 8 Heterocyclo)-C 1 ~C 10 Alkylene-,-C 1 ~C 10 Alkylene -C(=O)-, -C 1 ~C 10 Alkylene-C(O)NH-C 1 ~C 8 Alkylene-[O-CH 2 -CH 2 ] n -C(O)- (where n is between 1 and 26), C 1 ~C 10 Heteroalkylenes -C(=O)-, -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C(=O)-(where b is between 1 and 26), -(CH 2 -O-CH 2 ) b -C 1 ~C 8 Alkylene-C(=O)-(where b is 1 to 26), -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C 1 ~C 8 Alkylene-C(=O)-(where b is 1 to 26), -C 3 ~C 8 Carbocyclo-C(=O)-,-O-(C 1 ~C 8 Alkyl)-C(=O)-,-Arylene-C(=O)-,-C 1 ~C 10 Alkylene-arirene-C(=O)-, -arirene-C 1 ~C 10 Alkylene -C(=O)-, -C 1 ~C 10 Alkylene-(C) 3 ~C 8 Carbocyclo)-C(=O)-,-(C 3 ~C 8 Carbocyclo)-C 1 ~C 10 Alkylene -C(=O)-, -C 3 ~C 8 Heterocyclo-C(=O)-, -C 1 ~C 10 Alkylene-(C) 3 ~C 8 Heterocyclo)-C(=O)-,-(C 3 ~C 8 Heterocyclo)-C 1 ~C 10 Alkylene -C(=O)-, -C 1 ~C 10 Alkylene-NH-,-C 1 ~C 10 Heteroalkylene -NH-, -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -NH-(where b is 1 to 26), -(CH 2 -O-CH 2 ) b -C 1 ~C 8 Alkylene-NH-(where b is 1 to 26), -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C 1 ~C 8 Alkylene-NH-(where b is 1 to 26), -C 1 ~C 8 Alkylene-(C(=O))-NH-(CH 2 -O-CH 2 ) b -C(=O)-(where b is between 1 and 26), -C 1 ~C 8 Alkylene-(C(=O))-NH-(CH 2 -O-CH 2 ) b -C 1 ~C 8 Alkylene-C(=O)-(where b is 1 to 26), -C 1 ~C 8 Alkylene-NH-(C(=O))-(CH 2 -O-CH 2 ) b -NH- (where b is between 1 and 26), -C 1 ~C 8 Alkylene-NH-(C(=O))-(CH 2 -O-CH 2 ) b -C 1 ~C 8 Alkylene-NH-(where b is 1 to 26), -C 3 ~C 8 Carbocyclo-NH-,-O-(C 1 ~C 8 Alkyl)-NH-,-Arylene-NH-,-C 1 ~C 10 Alkylene-arylene-NH-,-arylene-C 1 ~C 10 Alkylene-NH-,-C 1 ~C 10 Alkylene-(C) 3 ~C 8 Carbocyclo)-NH-,-(C 3 ~C 8 Carbocyclo)-C 1 ~C 10 Alkylene-NH-,-C 3 ~C 8 Heterocyclo-NH-,-C 1 ~C 10 Alkylene-(C) 3 ~C 8 Heterocyclo)-NH-,-(C 3 ~C 8 Heterocyclo)-C 1 ~C 10 Alkylene-NH-,-C 1 ~C 10 Alkylene-S-,C 1 ~C 10 Heteroalkylene -S-, -C 3 ~C 8 Carbocyclo-S-,-O-(C 1 ~C 8 Alkyl)-S-,-Arylene-S-,-C 1 ~C 10 Alkilen-Arirene-S-,-Arirene-C 1 ~C 10 Alkylene-S-,-C 1 ~C 10 Alkylene-(C) 3 ~C 8 Carbocyclo)-S-,-(C 3 ~C 8 Carbocyclo)-C 1 ~C 10 Alkylene-S-,-C 3 ~C 8 Heterocyclo-S-,-C 1 ~C 10 Alkylene-(C) 3 ~C 8 Heterocyclo)-S-, or -(C 3 ~C 8 Heterocyclo)-C 1 ~C 10 It is alkylene-S-; or The stretcher group is maleimide (C 1 ~C 10 Alkylene-C(O)-, Maleimide(CH 2 OCH 2 ) p2 (C 1 ~C 10 Alkylene)C(O)-, Maleimide(C 1 ~C 10 Alkilen) (CH 2 OCH 2 ) p2 This includes C(O)- or their ring-opened forms, where p2 is 1 to 26; * indicates binding to the aforementioned binding substance, and the wavy line indicates the binding site to the enzyme-cleavable group. The composite according to claim 71.

73. The stretcher group is as follows: Derived from a more preferred structure, During the ceremony, wavy line This indicates the binding site of the stretcher group to the enzyme-cleavable group, The binding site to the aforementioned binding substance is located on the maleimide, on the primary amine, or on the alkyne functional group. The composite according to claim 72.

74. The linker compound has the following structure: Having one of the following, In the formula, the H on the benzyl OH may be replaced by a bond to one of the drug units or to the linking group bonded to at least one of the drug units. The composite according to any one of claims 26 to 30.

75. The complex according to any one of claims 30 to 74, wherein at least one of the drug units is bound to the linker compound at the binding site δ to form a drug-linker compound, which can then bind to the binding substance to form the complex.

76. The complex according to claim 75, wherein the drug unit is selected from cytotoxic substances, immunomodulators, nucleic acids, growth inhibitors, PROTACs, toxins, radioisotopes, and chelate ligands.

77. The complex according to claim 76, wherein the drug unit is a cytotoxic substance.

78. The complex according to claim 77, wherein the cytotoxic substance is selected from the group consisting of auristatin, meitansinoid, camptothecin, duocalmycin, and calicheamycin.

79. The complex according to claim 78, wherein the cytotoxic substance is MMAE, MMAF, exatecan, RS-exatecan, SS-exatecan, SN-38, DxD, meitansin, meitansinol, or ansamatosin-2.

80. The complex according to claim 78, wherein the cytotoxic substance is MMAE.

81. The complex according to claim 78, wherein the cytotoxic substance is exatecan.

82. The complex according to claim 76, wherein the drug unit is an immunomodulator.

83. The complex according to claim 82, wherein the immunomodulator is selected from a TRL7 agonist, a TLR8 agonist, a STING agonist, or a RIG-I agonist.

84. The complex according to claim 83, wherein the immunomodulator is a TLR7 agonist.

85. The complex according to claim 84, wherein the TLR7 agonist is imidazoquinoline, imidazoquinolineamine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzoimidazole-2-amine, tetrahydropyridopyrimidine, heteroalothiadiazide-2,2-dioxide, benzonaphthyridine, guanosine analog, adenosine analog, thymidine homopolymer, ssRNA, CpG-A, polyG10, or polyG3.

86. The complex according to claim 84, wherein the immunomodulator is a TLR8 agonist.

87. The complex according to claim 86, wherein the TLR8 agonist is selected from imidazoquinoline, thiazoloquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzoimidazole-2-amine, tetrahydropyridopyrimidine, or ssRNA.

88. The complex according to claim 83, wherein the immunomodulator is a STING agonist.

89. The complex according to claim 83, wherein the immunomodulator is a RIG-I agonist.

90. The composite according to claim 89, wherein the RIG-I agonist is selected from KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400, and KIN2000.

91. The complex according to claim 76, wherein the drug unit is a chelate ligand.

92. The composite according to claim 91, wherein the chelate ligand is selected from platinum (Pt), ruthenium (Ru), rhodium (Rh), gold (Au), silver (Ag), copper (Cu), molybdenum (Mo), titanium (Ti), or iridium (iridum) (Ir); radioactive isotopes, for example, yttrium-88, yttrium-90, technetium-99, copper-67, rhenium-188, rhenium-186, gallium-66, gallium-67, indium-111, indium-114, indium-115, lutetium-177, strontium-89, samarium-153, and lead-212.

93. The aforementioned drug-linker has the following structure: The composite according to claim 75, having one of the following.

94. Average drug loading (p load The composite according to any one of claims 26 to 93, wherein the ratio of the numbers is approximately 1 to approximately 8, approximately 2, approximately 4, approximately 6, approximately 8, approximately 10, approximately 12, approximately 14, approximately 16, approximately 3 to approximately 5, approximately 6 to approximately 8, or approximately 8 to approximately 16.

95. average p load The composite according to claim 93, wherein the ratio is approximately 8.

96. below: More selected, During the ceremony, Ab is the aforementioned binding substance, n is p load That is, The composite according to any one of claims 26 to 95.

97. The following structure: It has, During the ceremony, Ab is the aforementioned binding substance, n is p load And here, p load The ranges are approximately 1 to 8, 2, 4, 6, 8, 10, 12, 14, 16, 3 to 5, 6 to 8, or 8 to 16. The composite according to claim 26.

98. The following structure: It has, During the ceremony, Ab is the binding substance according to claim 4, n is p load And here, p load The ranges are approximately 1 to 8, 2, 4, 6, 8, 10, 12, 14, 16, 3 to 5, 6 to 8, or 8 to 16. The composite according to claim 26.

99. The following structure: It has, During the ceremony, Ab is the binding substance according to claim 21, n is p load And here, p load It is 8. The composite according to claim 26.

100. A pharmaceutical composition comprising a complex according to any one of claims 26 to 99 and a pharmaceutically acceptable carrier.

101. A method for treating PTK7+ cancer, comprising the step of administering a therapeutically effective amount of a binding substance according to any one of claims 1 to 21, a pharmaceutical composition according to claim 22, a complex according to any one of claims 26 to 99, or a pharmaceutical composition according to claim 100 to a subject in need of such treatment.

102. The method according to claim 101, wherein the PTK7+ cancer is a solid tumor or a hematological malignancy.

103. The method according to claim 102, wherein the PTK7+ cancer is selected from breast cancer (BC), lung cancer (LC), ovarian cancer (OVCA), esophageal cancer (EsC), gastric cancer (GC), bladder cancer (BLC), endometrial cancer (EC), head and neck cancer (HNC), cervical cancer, pharyngeal cancer, stomach cancer, myeloma, uterine cancer, colon cancer, hepatocellular carcinoma, and colorectal cancer.

104. The method according to claim 102, wherein the PTK7+ cancer is a hematological malignancy.

105. The method according to claim 102, wherein the PTK7+ cancer is a solid tumor.

106. The method according to claim 102, wherein the PTK7+ cancer is triple-negative breast cancer (TNBC).

107. The method according to claim 102, wherein the PTK7+ cancer is lung cancer.

108. The method according to claim 102, wherein the PTK7+ cancer is head and neck squamous cell carcinoma (HNSCC).

109. The method according to claim 102, wherein the PTK7+ cancer is non-small cell lung cancer (NSCLC).

110. The method according to claim 102, wherein the PTK7+ cancer is ovarian cancer.

111. The method according to claim 102, wherein the PTK7+ cancer is pharyngeal cancer.

112. The method according to claim 102, wherein the PTK7+ cancer is gastric cancer.

113. The method according to claim 102, wherein the PTK7+ cancer is endometrial cancer.

114. The method according to claim 102, wherein the PTK7+ cancer is endometrial adenocarcinoma.

115. The method according to claim 102, wherein the PTK7+ cancer is bladder cancer.

116. The method according to claim 102, wherein the PTK7+ cancer is a bladder transitional epithelial papilloma.

117. The method according to claim 102, wherein the PTK7+ cancer is esophageal cancer.

118. The method according to claim 117, wherein the esophageal cancer is esophageal squamous cell carcinoma (ESCC).

119. The method according to claim 102, wherein the PTK7+ cancer is gastroesophageal cancer.

120. The method according to claim 102, wherein the PTK7+ cancer is urothelial carcinoma.

121. The method according to claim 102, wherein the PTK7+ cancer is uterine cancer.

122. The method according to any one of claims 99 to 121, further comprising the step of administering immunotherapy to the subject.

123. The method according to claim 122, wherein the immunotherapy comprises a checkpoint inhibitor.

124. The method according to claim 123, wherein the checkpoint inhibitor is selected from antibodies that specifically bind to human PD-1, human PD-L1, or human CTLA4.

125. The method according to claim 123, wherein the checkpoint inhibitor is pembrolizumab, nivolumab, semiprimab, or ipilimumab.

126. The method according to any one of claims 101 to 125, further comprising the step of administering chemotherapy to the subject.

127. The method according to any one of claims 101 to 126, wherein the complex or pharmaceutical composition is administered intravenously.

128. The method according to claim 127, wherein the complex or pharmaceutical composition is administered in a dose of about 0.1 mg / kg to about 12 mg / kg.

129. The method according to any one of claims 101 to 123, wherein the treatment outcome of the subject is improved.

130. The method according to claim 129, wherein the improved treatment outcome is an objective response selected from disease stability, partial response, or complete response.

131. The method according to claim 130, wherein the improved treatment outcome is a reduction in tumor volume.

132. The method according to claim 130, wherein the improved treatment outcome is progression-free survival or disease-free survival.

133. Use of the complex according to any one of claims 26 to 99 or the pharmaceutical composition according to claim 100 for the treatment of PTK7+ cancer in a subject.

134. A method for treating an autoimmune disease, comprising the step of administering a therapeutically effective amount of the complex described in any one of claims 26 to 99 or the pharmaceutical composition described in claim 100 to a subject in need.

135. The method according to claim 134, wherein the autoimmune disease is rheumatoid arthritis, multiple sclerosis, or systemic lupus erythematosus.

136. The method according to any one of claims 134 to 135, further comprising the step of administering immunosuppressive therapy to the subject.

137. The method according to any one of claims 134 to 136, wherein the complex or pharmaceutical composition is administered intravenously.

138. The method according to claim 137, wherein the complex or pharmaceutical composition is administered in a dose of about 0.1 mg / kg to about 12 mg / kg.

139. The method according to any one of claims 134 to 138, wherein the treatment outcome of the subject is improved.

140. The method according to claim 139, wherein the improved treatment outcome is a reduction in disease progression or an alleviation of disease severity.

141. Use of the complex according to any one of claims 26 to 99 or the pharmaceutical composition according to claim 100 for the treatment of autoimmune diseases in subjects.

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