Linkers, drug linkers, and conjugates thereof, and methods of using same - Patent Application 20070122997

Abstract

[0003] Provided are linker compounds, or stereoisomers or salts thereof, comprising: (a) a linker unit having 1 to 4 attachment sites per drug unit and having one of the following structures (i) or (ii): (b) at least one polar group containing a polymer unit, optionally a sugar unit, optionally a carboxyl unit, or a combination thereof; and (c) optionally a Stretcher group having an attachment site for a targeting group, wherein α- is the attachment site to the enzyme-cleavable group; β- is the attachment site to the at least one polar group; and γ- is H, the attachment site to at least one of the drug units, or the attachment site to a linking group attached to at least one of the drug units. Drug-linker compounds and conjugates formed from the linker compounds, as well as related pharmaceutical compositions and methods, are also provided. TIFF2026504840000981.tif91128

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to International Application No. PCT / CN2023 / 071778, filed January 11, 2023, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] background There has been considerable interest in using monoclonal antibodies (mAbs) in the form of antibody-drug conjugates (ADCs) for the targeted delivery of cytotoxic agents to disease-related cells, such as cancer cells and other cells. The design of antibody-drug conjugates by attaching cytotoxic agents, immunomodulators, or other agents (collectively, "drugs") to antibodies, typically via linkers, requires consideration of various factors. These factors include the identification and positioning of chemical groups for drug attachment, the mechanism of drug release, the structural elements (if any) that result in drug release, and the structural changes, if any, 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 internalization of the antibody, the structural elements and mechanism of drug release must be consistent with the intracellular trafficking of the conjugate.

[0003] Another important factor in the design of antibody-drug conjugates is the amount of drug that can be delivered per targeting group (i.e., the number of drugs attached to each targeting group (e.g., antibody), referred to as drug content or drug loading). Historically, higher drug loadings were considered superior to lower drug loadings (e.g., 8-drug loading vs. 4-drug loading). The rationale was that higher conjugate loadings would deliver more drug (e.g., cytotoxic agents) to target cells. This rationale was supported by the observation that conjugates with higher drug loadings were more active against cell lines in vitro. However, subsequent studies revealed that this hypothesis was not confirmed in animal models. Conjugates with a drug loading of 4 or 8 of a particular auristatin were observed to have similar activity in mouse models. See, e.g., Hamblett et al., Clinical Cancer Res. 10:7063-70 (2004). Hamblett et al. further reported that ADCs with higher drug loadings were cleared from the circulation more rapidly in animal models. This more rapid elimination suggests that high-load species may be PK-challenged compared to low-load species. See Hamblett et al. Furthermore, higher-load conjugates have a lower maximum tolerated dose (MTD) in mice, resulting in a narrower reported therapeutic index. Id. In contrast, ADCs with a drug loading of 2 at engineered sites within monoclonal antibodies have been reported to have identical or superior PK and therapeutic indexes compared to certain ADCs with a loading of 4. See, e.g., Junutula et al., Clinical Cancer Res. 16:4769 (2010). Thus, the recent trend is to develop ADCs with lower drug loading.

[0004] Thus, there is a need for antibody-drug conjugate formats (and more broadly for formats for other conjugates) that allow for higher drug loadings while maintaining other properties of low-loading conjugates, such as favorable PK properties. Surprisingly, the present invention addresses these needs. [Prior art documents] [Non-patent literature]

[0005] [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) Summary of the Invention

[0006] overview Provided herein are linkers with hydrophilic characteristics that maintain the inherent properties of antibodies conjugated with linkers and drugs. In particular, the linkers help maintain the hydrophilic properties of antibodies when conjugated with higher drug loadings and / or hydrophobic drugs and other agents. Also provided are drug-linkers, conjugates containing linkers, and methods of using such conjugates for the treatment of cancer and other diseases.

[0007] In some embodiments, (a) having 1 to 4 binding sites per drug unit and having the following structure (i) or (ii): a linker unit with one of TIFF2026504840000002.tif39128; (b) at least one polar group comprising a polymer unit, optionally a sugar unit, optionally a carboxyl unit, or a combination thereof; and (c) optionally, a stretcher group having a binding site for a targeting group; or a stereoisomer or salt thereof, is provided, During the ceremony, α- is the attachment site to the enzyme-cleavable group; β- is the site of attachment to at least one polar group; δ- is H, a site of attachment to at least one of the Drug units, or a site of attachment to a linking group attached to at least one of the Drug units; the polymer units comprise polyamides, polyethers, or combinations thereof, wherein the polyethers comprise hydroxyl groups, polyhydroxyl groups, sugar groups, carboxyl groups, or combinations thereof; Each R a are independently H or C1-C6 alkyl; Each R b is independently halo, C 1~6 alkyl, a bond to at least one of the drug units, or a bond to at least one polar group; x is 0, 1, 2, 3, or 4; y is 0, 1, 2, or 3; R c is a bond, -C(O)-, -S(O)-, -SO2-, 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)-, -SO2-C1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, triazolyl-containing group, or a combination thereof.

[0008] In some embodiments, a Drug-Linker Compound is provided that comprises a linker compound described herein having at least one Drug unit attached via a linking moiety.

[0009] In some embodiments, a conjugate is provided that includes a targeting group attached to a drug-linker compound described herein.

[0010] In some aspects, a pharmaceutical composition is provided comprising a conjugate described herein and a pharmaceutically acceptable carrier.

[0011] In some aspects, provided are methods of treating a subject in need thereof, comprising administering to the subject a conjugate described herein or a pharmaceutical composition described herein, wherein the subject has cancer or an autoimmune disease, and wherein the conjugate binds to a target antigen associated with the cancer or autoimmune disease.

[0012] These and other aspects of the present invention will be more fully understood by reference to the following detailed description, non-limiting examples of specific embodiments, and the accompanying drawings. [Brief explanation of the drawings]

[0013] [Figure 1] 1 is a graph comparing tumor volume and treatment time for in vivo testing of antitumor agents against OVCAR-3. [Figure 2] 1 is a graph comparing tumor volume and treatment time for in vivo testing of antitumor agents against PA-1. [Figure 3] 1 is a graph comparing tumor volume and treatment time for in vivo testing of antitumor agents against NCI-H292. [Figure 4] 1 is a graph comparing tumor volume and treatment time for in vivo testing of antitumor agents against MDA-MB-468. [Figure 5] 1 is a graph comparing mean fluorescence intensity versus concentration for in vitro mAb and ADC binding to OVCAR-3 cells. [Figure 6] 1 is a graph comparing cell viability and concentration for ADCs against OVCAR-3 cells. [Figure 7] 1 is a graph comparing tumor volume and treatment time for in vivo testing of antitumor agents against NCI-H441. [Figure 8]1 is a graph comparing tumor volume and treatment time for in vivo testing of antitumor agents against HCC4006. [Figure 9] 1 is a graph comparing mean fluorescence intensity versus concentration for in vitro mAb and ADC binding to CFPAC-1 cells. [Figure 10] 1 is a graph comparing mean fluorescence intensity and concentration for in vitro mAb and ADC binding to PC-3 cells. [Figure 11] 1 is a graph comparing survival rates and concentrations of ADCs against CFPAC-1. [Figure 12] 1 is a graph comparing survival rates and concentrations for ADCs against PC-3. [Figure 13] 1 is a graph comparing tumor volume and treatment time for in vivo testing of antitumor agents against CFPAC-1. [Figure 14] 1 is a graph comparing tumor volume and treatment time for in vivo testing of antitumor agents against PC-3. DETAILED DESCRIPTION OF THE INVENTION

[0014] definition For convenience, certain terms in the specification, examples, and claims are defined here. Unless otherwise stated or implied from context, the following terms and phrases have the meanings provided below. The definitions are provided to aid in describing particular embodiments and are not intended to limit the claimed invention, as the scope of the invention is limited only by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0015] As used herein, and unless otherwise indicated, the terms "a" and "an" mean "one," "at least one," or "one or more." Unless otherwise required by context, singular terms used herein shall include pluralities and plural terms shall include the singular.

[0016] Unless the context otherwise requires, throughout the specification and claims, the words "comprises," "including," and the like are to be construed in their inclusive sense, i.e., "including, but not limited to," rather than in their exclusive or exhaustive sense.

[0017] The terms "decreased," "reduced," "reduced," "reduction," "reduce," and "inhibit" are generally used herein to mean a decrease by a statistically significant amount relative to a baseline value.

[0018] The terms "increased," "increase," or "enhance" or "activate" are generally used herein to mean an increase by a statistically significant amount relative to a baseline value.

[0019] As used herein, the terms "protein" and "polypeptide" are used interchangeably to refer to a series of amino acid residues linked together by peptide bonds between the alpha-amino and carboxyl groups of adjacent residues. The terms "protein" and "polypeptide" also refer to polymers of amino acids containing modified amino acids (e.g., phosphorylated, glycosylated, glycosylated, etc.) and amino acid analogs, regardless of their size or function. Although the terms "protein" and "polypeptide" are used to refer to relatively large polypeptides, while the term "peptide" is used to refer to small polypeptides, their usage in the art overlaps. The terms "protein" and "polypeptide" are used interchangeably herein to refer to encoded gene products and fragments thereof. Thus, exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments, and other equivalents, variants, fragments, and analogs of the foregoing.

[0020] As used herein, "epitope" refers to the amino acids normally bound by an immunoglobulin VH / VL pair, such as the antibodies, antigen-binding portions thereof, and other binding agents described herein. Other binding agents include non-antibody scaffolds. Epitopes can be formed on polypeptides from contiguous amino acids or non-contiguous amino acids arranged by tertiary folding of the protein. Epitopes formed from contiguous amino acids are typically retained upon exposure to denaturing solvents, while epitopes formed by tertiary folding are typically lost upon treatment with denaturing solvents. An epitope typically comprises at least three, more usually at least five, about nine, or about eight to ten 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 for the antibody, its antigen-binding site, or other immunoglobulin-based binding agent. In the case of single-domain antibodies, an epitope represents the unit of structure bound by the variable domain alone.

[0021] As used herein, "specifically binds" means that a binding agent (e.g., an antibody or antigen-binding portion thereof) described herein specifically binds to an antigen. -5 M (10000 nM) or less, e.g., 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 As used herein, "specifically binds" refers to the ability of a molecule described herein (e.g., an antibody or antigen-binding portion thereof, or a non-antibody scaffold) to bind to a target with a KD of 10 M or less. -5 M (10000 nM) or less, e.g., 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12Specific binding also refers to the ability to bind to a target with a KD of M or less. Specific binding can be influenced, for example, by the affinity and avidity of the antibody, antigen-binding moiety, or other binding substance, and the concentration of the target polypeptide. Those skilled in the art can use any suitable method, such as titrating the antibody or binding substance in a suitable cell-binding assay, to determine appropriate conditions under which the antibodies, antigen-binding moieties, and other binding substances described herein selectively bind to a target molecule. A binding substance that specifically binds to a target molecule is not displaced by a dissimilar competitor. In certain embodiments, an antibody or its antigen-binding moiety, or other binding substance, is said to specifically bind to a target molecule when it preferentially recognizes its target molecule in a complex mixture of proteins and / or macromolecules. Specific binding can be influenced, for example, by the affinity and avidity of the antibody, antigen-binding moiety, or non-antibody scaffold, and the concentration of the target polypeptide. Those skilled in the art can use any suitable method, such as titrating the antibody or non-antibody scaffold in a suitable cell-binding assay, to determine appropriate conditions under which the antibodies, antigen-binding moieties, and non-antibody scaffolds described herein selectively bind to a target molecule. Molecules that specifically bind to a target molecule are not displaced by dissimilar competitors. In certain embodiments, an antibody or antigen-binding portion thereof, or a non-antibody scaffold, is said to specifically bind a target molecule when it preferentially recognizes that target molecule in a complex mixture of proteins and / or macromolecules.

[0022] Unless otherwise indicated, the term "alkyl," by itself or as part of another term, refers to a substituted or unsubstituted straight-chain 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 to 5, 1 to 8, or 1 to 10 carbon atoms, respectively. Examples are 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), 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 (-CH2CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH( These include 4-methyl-2-pentyl (-CH(CH)CHCH(CH)), 4-methyl-2-pentyl (-CH(CH)CHCH(CH)), 3-methyl-3-pentyl (-C(CH)(CHCH)), 2-methyl-3-pentyl (-CH(CHCH)CH(CH)), 2,3-dimethyl-2-butyl (-C(CH)CH(CH)), and 3,3-dimethyl-2-butyl (-CH(CH)C(CH)).

[0023] Unless otherwise indicated, the term "alkenyl," by itself or as part of another term, refers to an alkyl group having at least one site of unsaturation (i.e., a carbon-carbon sp 2refers to a C2-C8 substituted or unsubstituted straight or branched hydrocarbon 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).

[0024] Unless otherwise indicated, "alkynyl" by itself or as part of another term refers to a C2-C8 substituted or unsubstituted straight or branched chain hydrocarbon having at least one site of unsaturation (i.e., a carbon-carbon sp triple bond). Examples include, but are not limited to, acetylene and propargyl.

[0025] Unless otherwise indicated, "alkylene" refers to a saturated, branched or straight-chain hydrocarbon radical of from 1 to 8 carbon atoms having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkane. Typical alkylene radicals include, but are not limited to, methylene (-CH-), 1,2-ethyl (-CHCH-), 1,3-propyl (-CHCHCH-), 1,4-butyl (-CHCHCHCH-), and the like.

[0026] Unless otherwise indicated, "alkenylene" refers to an unsaturated, branched or straight-chain hydrocarbon radical of 2 to 8 carbon atoms and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkene. Typical alkenylene radicals include, but are not limited to, 1,2-ethylene (-CH=CH-).

[0027] Unless otherwise indicated, "alkynylene" refers to an unsaturated, branched, straight-chain, or cyclic hydrocarbon radical of 2 to 8 carbon atoms and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkyne. Typical alkynylene radicals include, but are not limited to, acetylene, propargyl, and 4-pentynyl.

[0028] Unless otherwise indicated, the term "heteroalkyl," by itself or as part of another term, refers to a substituted or unsubstituted, stable, straight- or branched-chain hydrocarbon, or combination thereof, that is saturated and has from 1 to 10, preferably from 1 to 3, heteroatoms selected from the group consisting of O, N, Si, and S, where the nitrogen and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be quaternized. The O, N, and S heteroatoms may be located at any interior position of the heteroalkyl group (i.e., as part of the backbone) or at the position at which the alkyl group is attached to the remainder of the molecule. The Si heteroatom may be located at any position of the heteroalkyl group, including the position at which the alkyl group is attached to the remainder of the molecule. Examples of heteroalkyl groups include: -CHCHOCH, -CHCHNHCH, -CHCHN(CH)CH, -CHSCHCH, CHCHS(O)CH, -CHCHS(O)CH, and -Si(CH). For example, up to two heteroatoms may be consecutive, such as -CH2NHOCH3 and CH2OSi(CH3)3. In some embodiments, a C1-C4 heteroalkyl has 1-4 carbon atoms and 1 or 2 heteroatoms, and a C1-C3 heteroalkyl has 1-3 carbon atoms and 1 or 2 heteroatoms.

[0029] Unless otherwise indicated, the terms "heteroalkenyl" and "heteroalkynyl," by themselves or as part of another term, refer to a substituted or unsubstituted, stable, straight- or branched-chain 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 are optionally oxidized and the nitrogen heteroatom is optionally quaternized. The O, N, and S heteroatoms may be located at any interior position of the heteroalkenyl or heteroalkynyl group (i.e., as part of the backbone) or at the position at which the alkyl group is attached to the remainder of the molecule. The Si heteroatom may be located at any interior position of the heteroalkenyl or heteroalkynyl group, including the position at which the alkyl group is attached to the remainder of the molecule.

[0030] Unless otherwise stated, the term "heteroalkylene," by itself or as part of another term, refers to a substituted or unsubstituted divalent group derived from heteroalkyl (discussed above), as exemplified by -CH2CH2SCH2CH2- and -CH2SCH2CH2NHCH2-. In some embodiments, a C1-C4 heteroalkylene has 1-4 carbon atoms and 1 or 2 heteroatoms, and a C1-C3 heteroalkylene has 1-3 carbon atoms and 1 or 2 heteroatoms. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini. Furthermore, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied.

[0031] Unless otherwise stated, the terms "heteroalkenylene" and "heteroalkynylene," by themselves or as part of another term, refer to a substituted or unsubstituted divalent group derived from heteroalkenyl or heteroalkynyl (discussed above). In some embodiments, a C2-C4 heteroalkenylene or heteroalkynylene has 1 to 4 carbon atoms. For heteroalkenylene and heteroalkynylene groups, heteroatoms can also occupy either or both of the chain termini. Furthermore, for heteroalkenylene and heteroalkynylene linking groups, no orientation of the linking group is implied.

[0032] Unless otherwise indicated, "C3-C8 carbocycle," by itself or as part of another term, refers 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 derived by the removal of one hydrogen atom from a ring atom of a parent ring system. Representative -C3-C8 carbocycles 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-cycloheptatrienyl, cyclooctyl, and cyclooctadienyl.

[0033] Unless otherwise indicated, "C3-C8 carbocyclo," by itself or as part of another term, refers to a substituted or unsubstituted C3-C8 carbocyclic group, as defined above, in which another hydrogen atom of the carbocyclic group is replaced with a bond (i.e., is divalent).

[0034] Unless otherwise indicated, "C3~C 10 "Carbocycle" by itself or as part of another term refers 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 derived by the removal of one hydrogen atom from a ring atom of a parent ring system. Representative -C3-C 10Carbocycles 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-cycloheptatrienyl, cyclooctyl, and cyclooctadienyl. 10 Carbocycles may further include fused cyclooctyne carbocycles, such as the fused cyclooctyne compounds disclosed in International Publication No. WO 2011 / 136645 (the disclosure of which is incorporated herein by reference), including BCN (bicyclo[6.1.0]nonyne) and DBCO (dibenzocyclooctyne).

[0035] Unless otherwise indicated, a "C3-C8 heterocycle," by itself or as part of another term, refers to a substituted or unsubstituted, monovalent, aromatic or non-aromatic monocyclic or bicyclic ring system having 3 to 8 carbon atoms (also referred to as ring members) and 1 to 4 heteroatoms independently selected from N, O, P, or S, derived by the removal of a single hydrogen atom from a ring atom of a parent ring system. One or more N, C, or S atoms in a heterocycle may be oxidized. A ring containing a heteroatom may be aromatic or non-aromatic. Unless otherwise specified, a heterocycle is attached 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, tetrahydrofuranyl, tetrahydropyranyl, benzofuranyl, benzothiophene, indolyl, benzopyrazolyl, pyrrolyl, thiophenyl (thiophene), furanyl, thiazolyl, imidazolyl, pyrazolyl, pyrimidinyl, pyridinyl, pyrazinyl, pyridazinyl, isothiazolyl, and isoxazolyl. Unless otherwise indicated, the term "heterocarbocycle" is synonymous with the terms "heterocycle" or "heterocyclo" as described herein.

[0036] Unless otherwise indicated, "C3-C8 heterocyclo" by itself or as part of another term refers to a substituted or unsubstituted C3-C8 heterocyclic group, as defined above, in which one of the hydrogen atoms of the carbocyclic group is replaced with a bond (i.e., is divalent).

[0037] Unless otherwise indicated, "aryl," by itself or as part of another term, means a substituted or unsubstituted monovalent carbocyclic aromatic hydrocarbon radical of 6 to 20 carbons (preferably 6 to 14 carbons) derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system. Some aryl groups are represented in the exemplary structures as "Ar." Typical aryl groups include, but are not limited to, radicals derived from benzene, substituted benzene, naphthalene, anthracene, biphenyl, and the like. An exemplary aryl group is the phenyl group.

[0038] Unless otherwise indicated, "arylene," by itself or as part of another term, refers to an unsubstituted or substituted aryl group, as defined above, in which one of the aryl group's hydrogen atoms is replaced with a bond (i.e., it is divalent) and may be in the ortho, meta, or para orientation.

[0039] Unless otherwise indicated, "heteroaryl" and "heterocycle" refer to a ring system in which one or more ring atoms are heteroatoms, such as nitrogen, oxygen, and sulfur. The heterocycle radical contains 1 to 20 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S. The heterocycle may be a monocycle having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S), or a bicycle having 7 to 10 ring members (4 to 9 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, and S), such as a bicyclo[4,5], [5,5], [5,6], or [6,6] system.

[0040] Unless otherwise indicated, "heteroarylene," by itself or as part of another term, is an unsubstituted or substituted heteroaryl group, as defined above, in which one of the heteroaryl group's hydrogen atoms is replaced with a bond (i.e., it is divalent).

[0041] Unless otherwise indicated, "carboxyl" is COOH or COO - M + where M + is a cation.

[0042] Unless otherwise indicated, "oxo" refers to (C=O).

[0043] Unless otherwise indicated, "substituted alkyl" and "substituted aryl" refer to alkyl and aryl, respectively, in which one or more hydrogen atoms have each been independently replaced with a substituent. Exemplary substituents are -X, -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, where each X is independently a halogen: -F, -Cl, -Br, or -I; 10 are independently -H, -C1 to -C 20 Alkyl, -C6~C 20 Aryl, -C3~C 14 Heterocycle, protecting group or prodrug moiety. Typical substituents also include (=O). The alkylene, carbocycle, carbocyclo, arylene, heteroalkyl, heteroalkylene, heterocycle, and heterocyclo groups described above may also be substituted in the same manner.

[0044] Unless otherwise indicated, a "polyhydroxyl group" refers to an alkyl, alkylene, carbocycle, or carbocyclo group in which two or more, or three or more, hydroxyl groups have replaced hydrogen atoms on the carbon atoms of the carbon chain. In some embodiments, a polyhydroxyl group contains at least three hydroxyl groups. In some embodiments, a polyhydroxyl group contains carbon atoms with only one hydroxyl group per carbon atom. A polyhydroxyl group may contain one or more carbon atoms that are not substituted with a hydroxyl. A polyhydroxyl group may have each carbon atom substituted with a hydroxyl group. Examples of polyhydroxyl groups include linear (acyclic) or cyclic monosaccharides such as C6 or C5 sugars such as glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, gulose, idose, talose, aldose, and ketose; sugar acids such as gluconic acid, aldonic acid, uronic acid, or ulosonic acid; and amino sugars such as glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine. In some embodiments, the polyhydroxyl groups include disaccharides and polysaccharides in linear or cyclic form.

[0045] Unless otherwise indicated by context, "optionally substituted" refers to an alkyl, alkenyl, alkynyl, alkylaryl, arylalkylheterocycle, aryl, heteroaryl, alkylheteroaryl, heteroarylalkyl, or other substituent, moiety, or group as defined or disclosed herein, where a hydrogen atom of the substituent, moiety, or group is optionally replaced with a different moiety or group, or an alicyclic carbon chain comprising one of the substituents, moieties, or groups is interrupted by replacing a carbon atom in the chain with a different moiety or group. In some aspects, an alkene functional group replaces two consecutive sp3 carbon atoms of an alkyl substituent, with the proviso that the radical carbon of the alkyl moiety is not replaced, such that the optionally substituted alkyl is an unsaturated alkyl substituent.

[0046] Any substituents replacing hydrogen on any of the foregoing substituents, moieties, or groups are 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 -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(= and salts thereof, wherein each X is independently selected from the group consisting of a halogen atom, -F, -CI, -Br, and -I; and wherein each R" is independently selected from the group consisting of C1 to C6. 20 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, C6-C 24 Aryl, C3-C 24 Heterocyclyl (C5-C 24 or two of R" together with the heteroatom to which they are attached define a heterocyclyl; R' is hydrogen or R" where R" is a C1-C 20 Alkyl, C6-C 24 Aryl, C3-C 24 Heterocyclyl (C5-C 24 and protecting groups.

[0047] Typically, the optional substituents are -X, -OH, -OR", -SH, -SR", -NH, -NH(R), -NR'(R), -N(R), =NH, =NR, -CX, -CN, -NO, -NR'C(=O)H, NR'C(=O)R, -COH, -C(=O)H, -C(=O)R, -C(=O)NH, -C(=O)NR'R- and a salt thereof, wherein each X is independently selected from the group consisting of -F and -Cl, and R" is typically C1-C6 alkyl, C6-C7 alkyl, C8-C9 alkyl, C1-C6 alkyl, C1-C7 alkyl, C1-C6 ... 10 Aryl, C3-C 10 Heterocyclyl (C5-C 10 R' is independently selected from the group consisting of hydrogen, C1-C6 alkyl, C6 ... 10 Aryl, C3-C 10 Heterocyclyl (C5-C 10 (including heteroaryl), and protecting groups. More typically, the substituents are selected from the group consisting of -X, -R", -OH, -OR", -NH, -NH(R"), -N(R"), -N(R"), -CX, -NO, -NHC(=O)H, -NHC(=O)R", -C(=O)NH, -C(=O)NHR", -C(=O)N(R"), -COH, -COR", -C(=O)H, -C(=O)R", -C(=O)NH, -C(=O)NH(R"), -C(=O)N(R"), -C(=NR')NH, -C(=NR')NH(R"), -C(=NR')N(R"), protecting groups and salts thereof, wherein each X is -F and R" is independently C1-C6 alkyl, C6-C8 alkyl, C8-C9 alkyl, C9-C10 alkyl, C11-C12 alkyl, C12-C14 alkyl, C13-C15 alkyl, C14-C16 alkyl, C15-C16 alkyl, C16-C18 alkyl, C17-C18 alkyl, C18-C19 alkyl, C19-C20 alkyl, C19-C21 alkyl, C19-C22 alkyl, C19-C23 alkyl, C19-C24 alkyl, C19-C25 alkyl, C19-C26 alkyl, C19-C27 alkyl, C19-C28 alkyl, C19-C29 alkyl, C19-C29 alkyl, C19-C25 alkyl, C19-C26 ...9 alkyl, C19-C29 alkyl, 10 Aryl, C5-C 10 R' is selected from the group consisting of hydrogen, C1-C6 alkyl independently selected from R'' and a protecting group.

[0048] The compounds of the present invention or their pharmaceutically acceptable salts may contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be specified in terms of absolute stereochemistry as (R) or (S), or for amino acids, (D) or (L). The present invention is meant to include all such possible isomers, as well as their racemic and optically pure forms. 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, such as chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high-pressure liquid chromatography (HPLC). When the compounds described herein contain geometrically asymmetric olefinic double bonds or other centers, unless otherwise specified, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are intended to be included.

[0049] "Stereoisomers" refer to compounds composed of the same atoms joined by the same bonds but with different, incompatible three-dimensional structures. The present invention contemplates various stereoisomers and mixtures thereof, and includes enantiomers, which refer to two stereoisomers whose molecules are non-superimposable mirror images of one another. The present invention also includes "diastereomers," which refer to two or more stereoisomers of a compound that have different configurations at one or more of the equivalent stereocenters and that are not mirror images of each other.

[0050] Although the structures shown throughout this specification are depicted with particular stereocenters, this specification should be read to include variations of those stereocenters. For example, while the structure of exatecan may be depicted in the (S,S) configuration, it is contemplated that the (R,S) diastereomer of exatecan may be found in other embodiments of the conjugates described herein.

[0051] Unless otherwise indicated, the term "drug unit" or drug refers to cytotoxic agents (such as chemotherapeutic agents or drugs), immunomodulatory agents, nucleic acids (including siRNA), growth inhibitory agents, toxins (e.g., protein toxins, enzymatically active toxins of bacterial, fungal, plant or animal origin, or fragments thereof), radioisotopes, PROTACs, and other compounds that become active against target cells when delivered to them.

[0052] Unless otherwise indicated, the term "polymeric unit" refers to a polymeric portion made up of repeating subunits. Examples of polymeric units include polyamides and polyethers. In some embodiments, the polymeric unit is selected from an optionally substituted polyamide, a substituted polyether, or a combination thereof. In further embodiments, the polymeric unit is selected from: (i) Each R a are independently H or C 1~6 alkyl, and each R b are independently H or C 1~6 is alkyl, and n 0 are independently 2 to 26, TIFF2026504840000003.tif19128; an optionally substituted polyamide or a stereoisomer thereof; (ii) Each R b are independently H or C 1~6 is alkyl, and n 0 are independently 2 to 26, a substituted polyether containing TIFF2026504840000004.tif22128 or a stereoisomer thereof; or (iii) any combination thereof is selected from.

[0053] Unless otherwise indicated, the term "sugar unit" or "sugar group" refers to a carbohydrate group. Examples of sugar units include glycosides.

[0054] Unless otherwise indicated, the term "carboxyl unit" or "carboxyl group" refers to a group containing a carbonyl group [-C(O)-], a carboxyl group [-COH], and / or a carboxylate group [-COM, where M refers to a cationic counterion].

[0055] Unless otherwise indicated, the term "stretcher group" refers to the linking moiety that connects the targeting group to the enzyme-cleavable group.

[0056] Unless otherwise indicated, the term "polyamide" refers to a polymeric group composed of repeating subunits containing amide bonds.

[0057] Unless otherwise indicated, the term "polyether" refers to a polymeric group composed of repeating subunits containing ether linkages.

[0058] Unless otherwise indicated, the term "enzymatically cleavable group" refers to a group that is cleavable by the action of a metabolic process or reaction in an intracellular or extracellular environment, which cleaves the covalent bond between the Drug unit (e.g., a cytotoxic agent) and the Linker unit, or a portion thereof, to provide a free Drug unit or a Linker unit-Drug metabolite dissociated from the remainder of the Linker unit.

[0059] Unless otherwise indicated, the term "targeting group" refers to a macromolecule such as a protein, polypeptide, or peptide that specifically binds to a target molecule. Examples of targeting groups include antibodies.

[0060] As used herein, the phrase "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of a compound (e.g., a linker, a drug linker, or a conjugate). The compound typically contains at least one amino group and is therefore capable of forming an acid addition salt with this amino group. Exemplary salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, lineate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Pharmaceutically acceptable salts may involve the inclusion of other molecules, such as acetate, succinate, 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. When multiple charged atoms are part of a pharmaceutically acceptable salt, multiple counterions may be present. Thus, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counterions.

[0061] As used herein, the term "consisting essentially of" refers to elements required for a given embodiment. The term permits the presence of elements that do not materially affect the basic, novel, or functional characteristics of that embodiment.

[0062] As used herein, the term "consisting of" refers to the compositions, methods, and each component thereof described herein, excluding any element not recited in the description of the embodiment.

[0063] Except in the examples, or unless otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood to be modified in all instances by the term "about." When used in connection with percentages, the term "about" can mean + / - 1%.

[0064] The terms "statistically significant" or "significant" refer to statistical significance, which generally means a difference of two standard deviations (2 SD) above and below the reference value.

[0065] Other terms are defined herein within the description of various aspects of the invention.

[0066] Detailed Description Provided herein are linker compounds comprising a linker unit, at least one polar group, and optionally at least one of an enzyme-cleavable group and a stretcher group. Such linker compounds have hydrophilic characteristics that maintain the inherent properties of the linker and the antibody conjugated with the drug. In particular, the linker helps maintain the hydrophilic properties of the antibody at higher drug loadings and / or when conjugated with hydrophobic drugs and other agents.

[0067] In some embodiments, (a) having 1 to 4 binding sites per drug unit and having the following structure (i) or (ii): a linker unit with one of TIFF2026504840000005.tif38128; (b) at least one polar group comprising a polymer unit, optionally a sugar unit, optionally a carboxyl unit, or a combination thereof; and (c) optionally, a stretcher group having a binding site for a targeting group; or a stereoisomer or salt thereof, is provided, During the ceremony, α- is the attachment site to the enzyme-cleavable group; β- is the site of attachment to at least one polar group; δ- is H, a site of attachment to at least one of the Drug units, or a site of attachment to a linking group attached to at least one of the Drug units; the polymer units comprise polyamides, polyethers, or combinations thereof, wherein the polyethers comprise hydroxyl groups, polyhydroxyl groups, sugar groups, carboxyl groups, or combinations thereof; Each R a are independently H or C1-C6 alkyl; Each R b is independently halo, C 1~6 alkyl, a bond to at least one of the drug units, or a bond to at least one polar group; x is 0, 1, 2, 3, or 4; y is 0, 1, 2, or 3; R c is a bond, -C(O)-, -S(O)-, -SO2-, 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)-, -SO2-C1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, triazolyl-containing group, or a combination thereof.

[0068] In some embodiments, the linker unit has the following structure (ia), (ii-a), or (iii-a): TIFF2026504840000006.tif94128 or a stereoisomer or salt thereof.

[0069] In some embodiments, the linker unit has the following structure (ib), (ic), (id), (ie) or (if): TIFF2026504840000007.tif136132 or a stereoisomer or salt thereof.

[0070] In some embodiments, the linker unit has the following structure (ii-b) or (iii-b): TIFF2026504840000008.tif54128 or a stereoisomer or salt thereof.

[0071] polar group In some embodiments, there is provided a linker compound as described above, or a stereoisomer or salt thereof, wherein at least one polar group has the following formula: L3-N(CH2-(CH(XR)) k -X1(X2))2(X) or a stereoisomer or salt thereof, wherein each X is independently selected from NH and O; each R is independently selected from hydrogen, acetyl, monosaccharides, disaccharides, and polysaccharides; each X is independently selected from CH and C(O); each X is independently selected from H, OH, and OR; k is 1 to 10; and L is the point of attachment of the polar group to the remainder of the molecule.

[0072] In some embodiments, at least one sugar unit described above has the following structure (XII) or (XIII): TIFF2026504840000009.tif46128 or a stereoisomer or salt thereof, wherein each R is independently selected from hydrogen, monosaccharides, disaccharides, and polysaccharides; m is 1 to 8; and n is 0 to 4.

[0073] In some embodiments, the following: (a) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 (XX) or a stereoisomer, salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22are each independently a bond or a C1-C3 alkylene; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle, provided that R 24 and R 25 cannot both be H; n20 is 2 to 26; or (b) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 (XXI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene; R 24 and R 25 one of which is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;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-CH2-CH2-] n20 R 29 ] n21 -R 27 -NR 24 R 25 ] n27 (XXII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 26 and R 27 are arbitrary, and bonds, C1 to 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 to C 12 alkylene-NH-; R 24 and R 25 one of which is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;R 24 and R 25 the other is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 and polyethylene glycol, optionally having 1 to 24 ethylene glycol subunits; or -NR 24 R 25 together form a C3-C8 heterocycle, provided that R 24 and R 25 Both of R cannot be H; 29 is optional and independently selected from —C(O)—, —NH—, —C(O)—Ci-C6 alkylene-, —NH—Ci-C6 alkylene-, —Ci-C6 alkylene-NH—, —Ci-C6 alkylene-C(O)—, —NH(CO)—Ci-C6 alkylene-, —N(CH3)—(CO)—Ci-C6 alkylene-, —NH(CO)NH—, and triazole; n20 is 2 to 26; n21 is 1 to 4; and n27 is 1 to 4; or (d) ~R 20 -R 21 -[-C(R α )HC(O)-N(R N )-] n20 -R 22 -NR 24 R 25 (XXIII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21is a bond, 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 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)-; each R α are independently H or -R 22 -NR 24 R 25 and each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and;R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle, provided that R 24 and R 25 and 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond, C1-C3 alkylene, or -C1-C3 alkylene [O-CH2-CH2-] n20 and each R α are independently H or -R 22 -NR 24 R 25 and each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and;R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle, provided that R 24 and R 25 Both of them cannot be H;R 26 is H or C1-C4 alkyl; each n20 independently is 2 to 26, provided that at least one R α or R N Ha-R 22 -NR 24 R 25 is; or (f) ~R20 -R 21 -[C(R α )HC(O)-N(R N )-] n20 -R 22 -N-(R 23 -NR 24 R 25 )2(XXV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond, C1-C3 alkylene, or -C1-C3 alkylene-[O-CH2-CH2-] n20 and each R α are independently H or -R 22 -NR 24 R 25 and each R N are independently H or C1-C6 alkyl; each R 23 are independently C1-C6 alkylene; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle, provided that R 24 and R 25 cannot both be H; each n20 is independently 2 to 26;

[0023] Provided is a linker compound as described above, or a stereoisomer or salt thereof, comprising a polar group having a formula selected from:

[0074] In some embodiments, (a) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 (XX) or a stereoisomer, salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C1-C8 alkyl; a substituted -C(O)-C1-C8 alkyl; a chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 cannot both be H; n20 is 2 to 26; or (b) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 (XXI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene; R 24 and R 25one of which is selected from the group consisting of H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C1-C8 alkyl; a substituted -C(O)-C1-C8 alkyl; a chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;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-CH2-CH2-] n20 R 29 ] n21 -R 27 -NR 24 R 25 ] n27 (XXII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 26 and R 27 are arbitrary, and bonds, C1 to 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 to C 12 alkylene-NH-; R 24 and R 25 one of which is selected from the group consisting of H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C1-C8 alkyl; a substituted -C(O)-C1-C8 alkyl; a chelating agent; and R 28is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;R 24 and R 25 the other of which is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C1-C8 alkyl; a substituted -C(O)-C1-C8 alkyl; a chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 and polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, with the proviso that R 24 and R 25 Both of R cannot be H; 29 is optionally selected from —C(O)—, —NH—, —C(O)—Ci-C6 alkylene-, —NH—Ci-C6 alkylene-, —Ci-C6 alkylene-NH—, —Ci-C6 alkylene-C(O)—, —NH(CO)—Ci-C6 alkylene-, —N(CH3)—(CO)—Ci-C6 alkylene-, —NH(CO)NH—, and triazole; n20 is 2 to 26; n21 is 1 to 4; and n27 is 1 to 4; or (d) ~R 20 -R 21 -[-C(R α )HC(O)-N(R N )-] n20 -R 22 -NR 24 R 25 (XXIII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 is a bond, 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 22is 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 are independently H or -R 22 -NR 24 R 25 and each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and;R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C1-C8 alkyl; a substituted -C(O)-C1-C8 alkyl; a chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 and 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond, C1-C3 alkylene, or -C1-C3 alkylene [O-CH2-CH2-] n20 and each R α are independently H or -R 22 -NR 24 R 25 and each RN are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and;R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C1-C8 alkyl; a substituted -C(O)-C1-C8 alkyl; a chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 Both of them cannot be H;R 26 is H or C1-C4 alkyl; each n20 independently is 2 to 26, provided that 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 )HC(O)-N(R N )-] n20 -R 22 -N-(R 23 -NR 24 R 25 )2(XXV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond, C1-C3 alkylene, or -C1-C3 alkylene-[O-CH2-CH2-] n20 and each R α are independently H or -R 22 -NR 24 R 25 and each R N are independently H or C1-C6 alkyl; each R 23are independently C1-C6 alkylene; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C1-C8 alkyl; a substituted -C(O)-C1-C8 alkyl; a chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 cannot both be H; each n20 is independently 2 to 26; Provided is a linker compound as described above that includes a polar group having a formula selected from:

[0075] In some embodiments, the linker compound is R 24 and R 25 and R 24 and R 25 are each independently selected from H and a polyhydroxyl group, and R 24 and R 25 Both of them cannot be H.

[0076] In some embodiments, the polyhydroxyl group described above is a linear monosaccharide selected from C6 or C5 sugars, sugar acids, and amino sugars. In more specific embodiments, the polyhydroxyl group is a C6 or C5 sugar selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, gulose, idose, talose, aldose, and ketose; or the polyhydroxyl group is a sugar acid selected from gluconic acid, aldonic acid, uronic acid, and ulosonic acid; or the polyhydroxyl group is an amino sugar selected from glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine.

[0077] In some embodiments, the following: TIFF2026504840000010.tif95128TIFF2026504840000011.tif221120TIFF20265048400000 12.tif247148TIFF2026504840000013.tif215141TIFF2026504840000014.tif216158TIFF20 26504840000015.tif198158TIFF2026504840000016.tif224158TIFF2026504840000017.tif254158TIFF2026504840000018.tif167157, or a stereoisomer or salt thereof; wherein each R is independently H or alkyl; and each R 39 are independently selected from H, linear monosaccharides, and polyethylene glycols optionally having 1 to 24 ethylene glycol subunits; each n is independently 1 to 12; the wavy line represents R b or to an enzyme-cleavable group.

[0078] In some embodiments, R as described above 24 and R 25 One is a linear monosaccharide and the other is a cyclic monosaccharide.

[0079] In some embodiments, the following: TIFF2026504840000019.tif106128 or a stereoisomer or salt thereof, In the formula, R 41 is a cyclic monosaccharide; the wavy line is R b or to an enzyme-cleavable group.

[0080] In some embodiments, R as described above 24 and R 25 is independently a polyhydroxyl selected from cyclic monosaccharides, disaccharides, and polysaccharides.

[0081] In some embodiments, the following: A linker compound is provided, comprising a polar group selected from TIFF2026504840000020.tif188144 or a stereoisomer or salt thereof; In the formula, each R 45 is selected from H and a monosaccharide, a disaccharide, or a polysaccharide; R 46 is selected from cyclic monosaccharides, disaccharides, or polysaccharides; the wavy line represents R b or to an enzyme-cleavable group.

[0082] In some embodiments, R as described above 24 and R 25 is independently selected from a linear monosaccharide and a substituted linear monosaccharide, wherein the substituted linear monosaccharide is substituted with a monosaccharide, disaccharide, or polysaccharide.

[0083] In some embodiments, the following: Provided herein is a linker compound comprising a polar group selected from TIFF2026504840000021.tif179132 or a stereoisomer or salt thereof; In the formula, R 47 is a linear monosaccharide; each R 49 is selected from monosaccharides, disaccharides, and polysaccharides; the wavy line is R b or to an enzyme-cleavable group.

[0084] In some embodiments, R as described above 24 and R 25 are independently selected from linear monosaccharides and substituted monosaccharides, where the substituted linear monosaccharides are substituted with one or more substituents selected from alkyl, O-alkyl, aryl, O-aryl, carboxyl, ester, or amide, and may optionally be further substituted with a monosaccharide, disaccharide, or polysaccharide.

[0085] In some embodiments, R as described above 24 and R 25are independently selected from linear monosaccharides and substituted monosaccharides, where the substituted linear monosaccharides are substituted with one or more substituents selected from carboxyl, ester, and amide, and may be further substituted monosaccharides, disaccharides, or polysaccharides.

[0086] In some embodiments, the following: A linker compound is provided, comprising a polar group selected from TIFF2026504840000022.tif81128 or a stereoisomer or salt thereof; In the formula, each R 42 are independently selected from linear monosaccharides and substituted linear monosaccharides; each R 43 are independently selected from alkyl, O-alkyl, aryl, O-aryl, carboxyl, ester, and amide; the wavy line represents R b or to an enzyme-cleavable group.

[0087] In some embodiments, the following: TIFF2026504840000023.tif81128 or a stereoisomer or salt thereof, In the formula, each R 42 are independently selected from linear monosaccharides and substituted linear monosaccharides; each R 43 are independently selected from hydroxyl, carboxyl, ester, and amide; the wavy line represents R b or to an enzyme-cleavable group.

[0088] In some embodiments, R as described above 24 and R 25 is a -C(O)-polyhydroxyl group or a substituted -C(O)-polyhydroxyl group, and R 24 and R 25the other is H, -C(O)-polyhydroxyl, substituted -C(O)-polyhydroxyl, polyhydroxyl, or substituted polyhydroxyl; wherein the substituted -C(O)-polyhydroxyl and polyhydroxyl groups are substituted with monosaccharide, disaccharide, polysaccharide, alkyl, -O-alkyl, aryl, carboxyl, ester, or amide.

[0089] In some embodiments, R as described above 24 and R 25 is a -C(O)-polyhydroxyl group or a substituted -C(O)-polyhydroxyl group, and R 24 and R 25 the other is H, -C(O)-polyhydroxyl, substituted -C(O)-polyhydroxyl, polyhydroxyl, or substituted polyhydroxyl; wherein the substituted -C(O)-polyhydroxyl and polyhydroxyl groups are substituted with monosaccharides, disaccharides, polysaccharides, carboxyls, esters, or amides.

[0090] In some embodiments, the following: A linker compound is provided, comprising a polar group selected from TIFF2026504840000024.tif83128 or a stereoisomer or salt thereof; In the formula, the wavy line represents R b or to an enzyme-cleavable group.

[0091] In some embodiments, R as described above 24 and R 25 are independently selected from H, substituted -C1-C8 alkyl, substituted -C1-C4 alkyl, or substituted -C1-C3 alkyl; with the proviso that R 24 and R 25 cannot both be H; where substituted -C1-C8 alkyl, -C1-C4 alkyl, and -C1-C3 alkyl are substituted with hydroxyl and / or carboxyl.

[0092] In some embodiments, R as described above 24 and R 25are independently H or substituted -C1-C8 alkyl; provided that R 24 and R 25 cannot both be H; where the substituted -C1-C8 alkyl is substituted with hydroxyl and / or carboxyl.

[0093] In some embodiments, the following: A linker compound is provided, comprising a polar group selected from TIFF2026504840000025.tif228101 or a stereoisomer or salt thereof; In the formula, R 48 is selected from H, OH, CHOH, COOH, or -C1-C6 alkyl substituted with hydroxyl or carboxyl; the wavy line represents R b or to an enzyme-cleavable group.

[0094] In some embodiments, R as described above 24 and R 25 is selected from H, substituted —C(O)—C1-C8 alkyl, substituted —C(O)—C1-C4 alkyl, and substituted —C(O)—C1-C3 alkyl; R 24 and R 25 the other is selected from substituted -C(O)-C1-C8 alkyl, substituted -C(O)-C1-C4 alkyl, substituted -C(O)-C1-C3 alkyl, substituted -C1-C8 alkyl, substituted -C1-C4 alkyl, and substituted -C1-C3 alkyl, wherein the substituted -C(O)-C1-C8 alkyl, substituted -C(O)-C1-C4 alkyl, substituted -C(O)-C1-C3 alkyl, substituted -C1-C8 alkyl, -C1-C4 alkyl, and -C1-C3 alkyl are substituted with hydroxyl and / or carboxyl.

[0095] In some embodiments, R 24 and R 25 is H or substituted -C(O)-C1-C8 alkyl, and R 24 and R 25the other is a substituted -C(O)-C1-C8 alkyl or a substituted -C1-C8 alkyl, where the substituted -C(O)-C1-C8 alkyl and the substituted -C1-C8 alkyl are substituted with hydroxyl and / or carboxyl.

[0096] In some embodiments, the following: a linker compound comprising a polar group selected from TIFF2026504840000026.tif233121TIFF2026504840000027.tif21128 or a stereoisomer or salt thereof; In the formula, the wavy line represents R b or to an enzyme-cleavable group.

[0097] In some embodiments, R as described above 24 and R 25 is selected from H and optionally substituted aryl; provided that R 24 and R 25 Both of these cannot be H.

[0098] In some embodiments, the following: TIFF2026504840000028.tif64128 or a stereoisomer or salt thereof, In the formula, the wavy line represents R b or to an enzyme-cleavable group.

[0099] In some embodiments, R as described above 24 and R 25 are taken together to form an optionally substituted C3-C8 heterocycle or heteroaryl.

[0100] In some embodiments, the following structure: A linker compound is provided which comprises a polar group having TIFF2026504840000029.tif12128 or a stereoisomer or salt thereof, In the formula, the wavy line represents R b or to an enzyme-cleavable group.

[0101] In some embodiments, R as described above 24 and R 25 is independently selected from H and a chelating agent, wherein the chelating agent is selected from —NR by alkylene, arylene, carbocyclo, heteroarylene, or heterocarbocylo; 24 R 25 may be attached to the nitrogen of R 24 and R 25 cannot both be H. For example, in some embodiments, the chelator is selected from ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), triethylenetetraminehexaacetic acid (TTHA), benzyl-DTPA, 1,4,7,10-tetraazacyclododeca'e-''N'''',N'''-tetraacetic acid (DOTA), benzyl-DOTA, 1,4,7-triazacyclonona'e-''N',N''-triacetic acid (NOTA), benzyl-NOTA, 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA) and N,N'-dialkyl-substituted piperazines.

[0102] In some embodiments, the following: A linker compound is provided, comprising a polar group selected from TIFF2026504840000030.tif75128 or a stereoisomer or salt thereof; In the formula, the wavy line represents R b or to an enzyme-cleavable group.

[0103] In some embodiments, each monosaccharide listed above is independently selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, gulose, idose, talose, aldose, ketose, glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine; a sugar acid selected from gluconic acid, aldonic acid, uronic acid, and ulosonic acid; or an amino sugar selected from glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine.

[0104] In some embodiments, each of the above monosaccharides is independently selected from a C5 or C6 sugar selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, gulose, idose talose, aldose, and ketose; a sugar acid selected from gluconic acid, aldonic acid, uronic acid, and ulosonic acid; or an amino sugar selected from glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine.

[0105] In some embodiments, the above-described binding sites are formed from functional groups of precursor compounds of polar groups, where the functional groups are selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, and protected forms thereof.

[0106] In some embodiments, the following: (a) ~R 20 -R 21 -[O-CH2-CH2] n20 -R22 -R 30 (XXX) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 30 is an optionally substituted C3 to C 10 carbocycle; thiourea; optionally substituted thiourea; urea; optionally substituted urea; sulfamide; alkyl sulfamide; acylsulfamide; optionally substituted alkyl sulfamide; optionally substituted acylsulfamide; sulfonamide; optionally substituted sulfonamide; guanidine, including alkyl guanidine and aryl guanidine; phosphoramide; or optionally substituted phosphoramide; or R 30 represents azido, alkynyl, substituted alkynyl, -NH-C(O)-alkynyl, -NH-C(O)-alkynyl-R 65 cyclooctyne; selected from -NH-cyclooctyne, -NH-C(O)-cyclooctyne, or -NH-(cyclooctyne)2; where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (b) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NH-C(O)-R 31 (XXXI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22are each independently a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 R 35 is azide, alkynyl, alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (c) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -C(O)NH-R 31 (XXXII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch independently having 1 to 26 ethylene glycol subunits and each branch having R at its end 35 R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, and optionally substituted heteroaryl; n20 is 2 to 26; (d) ~R20 -R 21 -[O-CH2-CH2] n20 -R 22 -C(O)NR 31 -R 22 -NR 24 R 25 (XXXIII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 31 is H or R 22 -NR 24 R 25 and;R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 24 and R 25 Both of these cannot be H; n20 is 2 to 26; (e) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -N(R 33 -R 31 )2(XXXIV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 R 33is C1-C3 alkylene, C1-C3 alkylene-C(O), -C(O)-C1-C3 alkylene, or -C(O)-C1-C3 alkylene-C(O); R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (f) ~R 20 -(R 21 -[CH2-CH(OR 34 )-CH2-O] n20 -R 36 ) n25 (XXXV) or a stereoisomer or salt thereof, wherein R 20 is part R b or an enzyme-cleavable group; each R 21 are independently a bond, —O—, or a C1-C3 alkylene group; each R 34 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 R 25 and;R N is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; or a substituted polyhydroxyl group, with the proviso that R 24 and R 25 Both of R cannot be H; 36 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 are independently H or C1-C6 alkyl; R 44 and R 45 are each 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 Both of them will not become H. each n20 is independently 1 to 26; and n25 is 2 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 -R 25 (XXXVI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 , R 22 and R 23 are each independently a bond or a C1-C3 alkylene group; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R24 and R 25 each n20 independently represents 0 to 26, and each n21 independently represents 0 to 26, provided that at least one of n20 or n21 is 2 to 26; n22 is 1 to 5; and each n23 independently represents 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) (i) or a stereoisomer or salt thereof, wherein: R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R N is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; and a substituted polyhydroxyl group, with the proviso that R 44 and R 45 Both of R cannot be H; 34 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 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 are independently H or C1-C6 alkyl; R 44 and R 45 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 44 and R 45 n20 is 2 to 26; n21 is 1 to 26; n25 is 1; or ~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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R N is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; and a substituted polyhydroxyl group, with the proviso that R 24 and R 25 n20 is 2 to 26; n21 is 1 to 4; and 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond, 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 are independently H or -R 22 -NR 24 R 25 and each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and;R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle, provided that R 24 and R 25 each n20 independently is 0 to 26, provided that at least one n20 is 2 to 26; and n25 is 1 or 2; and (k) TIFF2026504840000031.tif23128 or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 , R 22 and R 23 each independently represents a bond, 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 are independently H or -R 22 -NR 24 R 25 and each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and;R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle, provided that R 24 and R 25 Both of them cannot be H;R 26is H or C1-C6 alkyl; each n20 independently is 0-26, provided that at least one n20 is 2-26; each n21 independently is 0-26, provided that at least one n21 is 2-26; Linker compounds are provided that include a polar group having a formula selected from:

[0107] In some embodiments, the following: (a) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -R 30 (XXX) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 30 is an optionally substituted C3 to C 10 carbocycle; thiourea; optionally substituted thiourea; urea; optionally substituted urea; sulfamide; alkyl sulfamide; acylsulfamide; optionally substituted alkyl sulfamide; optionally substituted acylsulfamide; sulfonamide; optionally substituted sulfonamide; guanidine, including alkyl guanidine and aryl guanidine; phosphoramide; or optionally substituted phosphoramide; or R 30 represents azido, alkynyl, substituted alkynyl, -NH-C(O)-alkynyl, -NH-C(O)-alkynyl-R 65 cyclooctyne; selected from -NH-cyclooctyne, -NH-C(O)-cyclooctyne, or -NH-(cyclooctyne)2; where R 65is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (b) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NH-C(O)-R 31 (XXXI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (c) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -C(O)NH-R 31 (XXXII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22are each independently a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch independently having 1 to 26 ethylene glycol subunits and each branch having R at its end 35 R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, and optionally substituted heteroaryl; n20 is 2 to 26; (d) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -C(O)NR 31 -R 22 -NR 24 R 25 (XXXIII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 31 is H or R 22 -NR 24 R 25 and;R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 24 and R 25 Both of these cannot be H; n20 is 2 to 26; (e) ~R 20 -R 21 -[O-CH2-CH2]n20 -R 22 -N(R 33 -R 31 )2(XXXIV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 R 33 is C1-C3 alkylene, C1-C3 alkylene-C(O), -C(O)-C1-C3 alkylene, or -C(O)-C1-C3 alkylene-C(O); R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (f) ~R 20 -(R 21 -[CH2-CH(OR 34 )-CH2-O] n20 -R 36 ) n25 (XXXV) or a stereoisomer or salt thereof, wherein R 20 is part R b or an enzyme-cleavable group; each R 21 are independently a bond, —O—, or a C1-C3 alkylene group; each R 34 are independently H, -[CH2-CH(OH)-CH2-O] n20 -R 36 , -C(O)-NR24 R 25 , or -C(O)N(R N )-C1~C6 alkylene-NR 24 R 25 and;R N is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; or a substituted polyhydroxyl group, with the proviso that R 24 and R 25 Both of R cannot be H; 36 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 are independently H or C1-C6 alkyl; R 44 and R 45 are each 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 Both of them will not become H. each n20 is independently 1 to 26; and n25 is 2 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 , R 22 and R 23 are each independently a bond or a C1-C3 alkylene group; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 24 and R 25 each n20 independently represents 0 to 26, and each n21 independently represents 0 to 26, provided that at least one of n20 or n21 is 2 to 26; n22 is 1 to 5; and each n23 independently represents 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) (i) or a stereoisomer or salt thereof, wherein: R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R N is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; and a substituted polyhydroxyl group, with the proviso that R 44 and R 45 Both of R cannot be H;34 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 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 are independently H or C1-C6 alkyl; R 44 and R 45 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 44 and R 45 n20 is 2 to 26; n21 is 1 to 26; n25 is 1; or ~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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; RN is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; and a substituted polyhydroxyl group, with the proviso that R 24 and R 25 n20 is 2 to 26; n21 is 1 to 4; and 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond, 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 are independently H or -R 22 -NR 24 R 25 and each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and;R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C(O)-C1-C8 alkyl; a chelating agent; R 28is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 each n20 independently is 0 to 26, provided that at least one n20 is 2 to 26; and n25 is 1 or 2; and (k) TIFF2026504840000032.tif23128 or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 , R 22 and R 23 each independently represents a bond, 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 are independently H or -R 22 -NR 24 R 25 and each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and;R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C(O)-C1-C8 alkyl; a chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 Both of them cannot be H;R 26is H or C1-C6 alkyl; each n20 independently is 0-26, provided that at least one n20 is 2-26; each n21 independently is 0-26, provided that at least one n21 is 2-26; Linker compounds are provided that include a polar group having a formula selected from:

[0108] In some embodiments, the following: ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NH-C(O)-R 31 (XXXI), ~R 20 -R 21 -[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 linker compound is provided that includes a polar group having a formula selected from: In the formula, R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 R 33 is C1-C3 alkylene, -C1-C3 alkylene-C(O), -C(O)-C1-C3 alkylene, or -C(O)-C1-C3 alkylene-C(O); R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; the wavy (~) line is R 20 indicates the binding site to; n20 is 2-26.

[0109] In some embodiments, the following: a linker compound is provided that includes a polar group formed from a precursor group selected from TIFF2026504840000033.tif219142; In the formula, R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; the wavy line represents R b or to an enzyme-cleavable group.

[0110] In some embodiments, R as described above b The attachment site to or to the enzyme-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, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, and protected forms thereof.

[0111] In some embodiments, the formula: ~R 20 -(R 43 -R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 )n42 (XL) or a stereoisomer or salt thereof, In the formula, R 20 is part R b or to an enzyme-cleavable group; R 41 and R 42 are each independently a bond or C1-C6 alkylene; 43 are independently bonded, C1 to C 12 Alkylene, -OC1~C 12 Alkylene, -C(=O)-, -NR a -C1~C 12 Alkylene, -C1~C 12 Alkylene-NR a -, -C(O)-C1~C 12 Alkylene, -C1~C 12 Alkylene-C(O)-, -NR a -C1~C 12 Alkylene -C(O)-, -C(O)-C1-C 12 Alkylene-NR a -, -NR a -C(O)-NR a -, -NR a -C(O)-, -NR a -C(O)-C1~C 12 Alkylene, -C(O)-NR a -C1~C 12 Alkylene, heteroarylene, heteroaryl-C1-C 12 Alkylene, heteroaryl-C1-C 12 Alkylene -C(O)- or -C(O)NR 46 R 47 wherein each alkylene is optionally substituted with hydroxyl, SO3H and / or oxo; R a is H, C1-C6 alkyl, polyhydroxyl group, or substituted polyhydroxyl group, and R 46 and R 47 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, wherein one of the C1-C2 alkylenes is NR44 R 45 is bonded to; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 cannot both be H; n40 is 2 to 26; n41 is 1 to 6; and n42 is 1 to 6.

[0112] In some embodiments, the formula: ~R 20 -(R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLI) or a stereoisomer or salt thereof, In the formula, R 20 is part R b or to an enzyme-cleavable group; R 41 and R 42 are each independently a bond or a C1-C6 alkylene; R 43 is a bond, C1~C 12 Alkylene, -OC1~C 12 Alkylene, -C(=O)-, -NR a -C1~C 12 Alkylene, -C1~C 12 Alkylene-NR a -, -C(O)-C1~C 12 Alkylene, -C1~C 12 Alkylene-C(O)-, -NR a -C1~C 12 Alkylene -C(O)-, -C(O)-C1-C 12 Alkylene-NR a -, -NR a -C(O)-NRa -, -NR a -C(O)-, -NR a -C(O)-C1~C 12 Alkylene, C(O)-NR a -C1~C 12 Alkylene, heteroarylene, heteroaryl-C1-C 12 Alkylene, heteroaryl-C1-C 12 Alkylene -C(O)-, and -C(O)NR 46 R 47 wherein each alkylene is optionally substituted with hydroxyl, SO3H and / or oxo; R a is H, C1-C6 alkyl, polyhydroxyl group, or substituted polyhydroxyl group, and R 46 and R 47 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, wherein one of the C1-C2 alkylenes is NR 44 R 45 is bonded to; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 cannot both be H; n40 is 1 to 26; n41 is 1 to 6; and n42 is 1 to 6.

[0113] In some embodiments, the formula: ~R 20 -(R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLII) or a stereoisomer or salt thereof, During the ceremony, R 20 is part R b or to an enzyme-cleavable group; R 41 and R 42 are each independently a bond or a C1-C3 alkylene; R 43 is a bond, C1-C6 alkylene, -OC1-C 12 Alkylene, -C(=O)-, -NR a -C1~C 12 Alkylene, -C1-C6 alkylene-NR a -, -C(O)-C1-C6 alkylene, -C1-C6 alkylene-C(O)-, -NR a -C1-C6 alkylene-C(O)-, -C(O)-C1-C6 alkylene-NR a -, -NR a -C(O)-NR a -, -NR a -C(O)-, -NR a -C(O)-C1-C6 alkylene, -C(O)-NR a -C1~C 12 Alkylene, -heteroarylene, heteroaryl-C1-C6 alkylene, heteroaryl-C1-C6 alkylene-C(O)-, and -C(O)NR 46 R 47 wherein each alkylene is optionally substituted with hydroxyl, SO3H and / or oxo; R a is H, C1-C6 alkyl, polyhydroxyl group, or substituted polyhydroxyl group, and R 46 and R 47 One of the groups is H or C1-C6 alkylene, and the other is C1-C 12 alkylene, wherein one of the C1-C2 alkylenes is NR 44 R 45 It is bound to; R 44 and R 45are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 cannot both be H; n40 is 1 to 16; n41 is 1 to 4; and n42 is 1 to 4.

[0114] In some embodiments, R as described above 20 is formed from a functional group of a 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, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, or a protected form thereof.

[0115] In some embodiments, R as described above 20 has the following structure: TIFF2026504840000034.tif84159TIFF2026504840000035.tif233151 or a stereoisomer thereof, wherein R is H, a C1-C6 alkyl, or a polyhydroxyl group, and n is 0 to 12; TIFF2026504840000036.tif4128 is R b indicates the site of attachment to or to an enzyme-cleavable group, TIFF2026504840000037.tif4128 shows the site of attachment of the polar group to the rest of the molecule.

[0116] In some embodiments, R as described above 20 has the following structure: TIFF2026504840000038.tif236161TIFF2026504840000039.tif65150 or a stereoisomer thereof, In the formula, n is 0 to 12; TIFF2026504840000040.tif4128 is R b indicates the site of attachment to or to an enzyme-cleavable group, TIFF2026504840000041.tif4128 shows the site of attachment of the polar group to the rest of the molecule.

[0117] In some embodiments, R as described above 43 -(NR 44 R 45 ) n41 has the following structure: TIFF2026504840000042.tif129148 or a stereoisomer thereof, In the formula, R a is H, C1-C6 alkyl, a polyhydroxyl group, or a substituted polyhydroxyl group; p is an integer from 1 to 6; TIFF2026504840000043.tif4128 is the R to the rest of the polar group 43 The binding site of

[0118] In some embodiments, R as described above 43 -(NR 44 R 45 ) n41 has the following structure: TIFF2026504840000044.tif157156 or a stereoisomer thereof, During the ceremony, TIFF2026504840000045.tif4128 is the R to the rest of the polar group 43 The binding site of

[0119] In some embodiments, the —NR 44 R 45 has the following structure: TIFF2026504840000046.tif57144TIFF2026504840000047.tif199160 or a stereoisomer thereof, During the ceremony, TIFF2026504840000048.tif4128 is the polar group -NR to the rest of the 44 R 45 The binding site of

[0120] In some embodiments, the linker unit has the following structure prior to attachment: TIFF2026504840000049.tif229155TIFF2026504840000050.tif237102TIFF2026504840000051.tif219115TIF F2026504840000052.tif253163TIFF2026504840000053.tif231163TIFF2026504840000054.tif230168TIFF202 6504840000055.tif187161TIFF2026504840000056.tif188162TIFF2026504840000057.tif205162TIFF202650 4840000058.tif225162TIFF2026504840000059.tif219141TIFF2026504840000060.tif191163TIFF2026504840 000061.tif220161TIFF2026504840000062.tif225169TIFF2026504840000063.tif241166TIFF2026504840000 064.tif182164TIFF2026504840000065.tif203145TIFF2026504840000066.tif220166TIFF2026504840000067. tif249168TIFF2026504840000068.tif246106TIFF2026504840000069.tif23578TIFF2026504840000070.tif197170TIFF2026504840000071.tif242170TIFF2026504840000072.tif154170, In the formula, (*) represents the site R bor an enzyme-cleavable group; each R is independently H or C1-C6 alkyl; R' is H, C1-C6 alkyl, -N(R 24 )(R 25 ), or —COH; each n is independently 1 to 12; X is O, NR, or —CH—; V is a bond or C1 to C6 alkyl; R 24 and R 25 one of which is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C(O)-C1-C8 alkyl; a chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;R 24 and R 25 the other is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C(O)-C1-C8 alkyl; a chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 and polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, with the proviso that R 24 and R 25 and R' are not both H. In some of the above embodiments, each R is independently H or C1-C6 alkyl; R' is H, C1-C6 alkyl, -N(R 24 )(R 25 ), or —COH; each n is independently 1 to 12; X is O, NR, or —CH—; V is a bond or C1 to C6 alkyl; R 24 and R 25 one of which is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;R 24 and R 25 the other is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 and polyethylene glycol, optionally having 1 to 24 ethylene glycol subunits; or -NR 24 R 25 together form a C3-C8 heterocycle, provided that R 24 and R 25 Both of them cannot be H.

[0121] In some embodiments, the following: ~R 40 -(R 43 -R 41 -[O-CH2-CH2] n40 -R 46 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLIII) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 41 and R42 are each independently a bond or C1-C6 alkylene; 43 are independently bonded, C1 to C 12 Alkylene, -OC1~C 12 Alkylene, -C(=O)-, -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-C 12 Alkylene, C1-C 12 Alkylene-NH-C(O)-, -heteroarylene, heteroaryl-C1-C 12 Alkylene, heteroaryl-C1-C 12 Alkylene -C(O)- or -C(O)NR 46 R 47 where R 46 and R 47 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 Both of R cannot be H; 46 is independently -NR 50 -, -NR 50 -C1~C6 alkylene-NR 50 -, -NR 50 -C(O)-NR 50 -S(O)2-NR 50 -, or -NR50 -C(O)-C 1~6 alkylene; each R 50 are independently selected from H, C1-C6 alkyl, or polyhydroxyl groups; each n40 is independently 2 to 26; n41 is 1 to 6; and n42 is 1 to 6; (b) ~R 40 -(R 51 -[O-CH2-CH2] n43 -R 52 -X1-R 55 -X2-R 53 -[O-CH2-CH2] n43 -R 54 -[X3-R 56 ] n44 -R 57 ) n45 (XLIV) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 51 , R 52 , R 53 and R 54 are each independently a bond or C1-C6 alkylene; X1, X2, and X3 are each independently -NR N -C(O)- or -C(O)-NR N - and each R N each independently represents H, a C1-C6 alkyl, or a polyhydroxyl group; R 55 and R 56 each independently represents a divalent polyhydroxyl group; R 57 is H, OH, or C1-C6 alkyl; each n43 independently is 0 to 26, provided that at least one n43 is 1 to 26; n44 is 0 to 10; n45 is 1 or 2; and (c) ~R 40 -R 51 -[O-CH2-CH2] n43 -R 52 -N-(R 53 -X1-R 54-[O-CH2-CH2] n43 -(NR 44 R 45 ))2 (XLV) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 51 , R 53 and R 54 are each independently a C1-C6 alkylene which may be bonded or substituted; R 52 is a bond, C1-C6 alkylene, —C(O)—, or —OC(O)—; each X1 is independently —NR N -C(O)- or -C(O)-NR N - and each R N each independently represents H, a C1-C6 alkyl, or a polyhydroxyl group; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 cannot both be H; each n43 is independently 2 to 26; Linker compounds are provided that include a polar group having a formula selected from:

[0122] In some embodiments, the enzyme-cleavable group has the following structure prior to attachment to the enzyme-cleavable group and / or to the linker unit: a linker compound is provided that includes a polar group having one of: TIFF2026504840000073.tif209140TIFF2026504840000074.tif56170; In the formula, (*) represents the site R b represents a site of attachment to a hydroxyl group or to an enzyme-cleavable group; each R is independently H, an alkyl, or a polyhydroxyl group; R 44 and R 45are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 and each n is independently 1 to 12.

[0123] In some embodiments, TIFF2026504840000075.tif214160 or a stereoisomer or salt thereof, wherein each Y is independently R 76 or TIFF2026504840000076.tif18128; each R 76 are independently H, acetyl, -P(=O)(OH)-, or -(CH2) v -OS(=O)2(OH); each R a and R b are independently H or R a and R b together with the carbon to which they are attached form an oxo group; each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; each v is independently 1 to 6; and each * is R b In some embodiments, Y is a bond to R 76 In another embodiment, Y is The file is TIFF2026504840000077.tif18128.

[0124] In some embodiments, TIFF2026504840000078.tif207161 or a stereoisomer or salt thereof, In the formula, each R 76 are independently H, acetyl, -P(=O)(OH) - , or -(CH2)v S(=O)2(OH); each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; each v is independently 1 to 6; and each * is R b or to an enzyme-cleavable group.

[0125] In some embodiments, TIFF2026504840000079.tif214159 or a stereoisomer or salt thereof, wherein each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; and each * is R b or to an enzyme-cleavable group.

[0126] In some embodiments, each R described above a and R b are independently H. In another embodiment, R a and R b together with the carbon to which they are attached form an oxo group.

[0127] In some embodiments, q described above is 10 to 20. For example, in some embodiments, q described above is 12.

[0128] In some embodiments, the following: TIFF2026504840000080.tif247158TIFF2026504840000081.tif235157TIFF2026504840000082.tif193157TIFF2026504840000083.tif 197157TIFF2026504840000084.tif255157TIFF2026504840000085.tif219157TIFF2026504840000086.tif211157TIFF20265048400000 87.tif204158TIFF2026504840000088.tif212157TIFF2026504840000089.tif235158TIFF2026504840000090.tif216141TIFF20265048 40000091.tif186157TIFF2026504840000092.tif254152TIFF2026504840000093.tif195148TIFF2026504840000094.tif205146TIFF20 26504840000095.tif253168TIFF2026504840000096.tif234144TIFF2026504840000097.tif202140TIFF2026504840000098.tif237148 TIFF2026504840000099.tif231145TIFF2026504840000100.tif229111TIFF2026504840000101.tif225120TIFF2026504840000102.tif 228158TIFF2026504840000103.tif201138TIFF2026504840000104.tif162159TIFF2026504840000105.tif199147TIFF2026504840000106.tif206159TIFF2026504840000107.tif248158TIFF2026504840000108.tif224157, or a stereoisomer or salt thereof; During the ceremony, each Z is joined at *, Independently selected from TIFF2026504840000109.tif124159, each TIFF2026504840000110.tif2128 is R b or to an enzyme-cleavable group.

[0129] In some embodiments, the compound of the formula: A linker compound is provided which comprises a polar group comprising at least one carboxyl unit having TIFF2026504840000111.tif28128 or a stereoisomer or salt thereof, During the ceremony, (a)L 70 is selected from C1-C8 alkylene, C1-C8 alkylene-C(O)-, -C(O)-C1-C8 alkylene-, and -C(O)-C1-C8 alkylene-C(O)-, * is R b to the enzymatically cleavable group or to the remainder of the polar group; R 70 Ha~NR 71 (R 72 -R 73 ), where R 71 is H, C1~C 12 Alkyl, substituted C1-C 12 alkyl, or polyethylene glycol (which may have 1 to 12 ethylene glycol subunits); R 72 is a bond or is selected from an optionally substituted C1-C3 alkylene, an optionally substituted ether, an optionally substituted thioether, an optionally substituted ketone, an optionally substituted amide, a polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), an optionally substituted carbocycle, an optionally substituted aryl, or an optionally substituted heteroaryl; R 73is a carboxyl or polycarboxyl, wherein the polycarboxyl contains 1 to 10, or 1 to 6, or 1 to 4 carboxyl groups, and the carboxyl groups are interconnected by alkyl, alkylene, substituted alkyl, substituted alkylene, heteroalkyl, heteroalkylene, amino, and / or amido; or (b)L 70 is selected from C1-C8 alkylene, C1-C8 alkylene-C(O)-, -C(O)-C1-C8 alkylene-, and -C(O)-C1-C8 alkylene-C(O)-, * is R b to the enzymatically cleavable group or to the remainder of the polar group; R 70 Ha~NR 71 (R 75 - (R 73 )2), where R 71 is H, C1~C 12 Alkyl, substituted C1-C 12 alkyl, or polyethylene glycol (which may have 1 to 12 ethylene glycol subunits); R 75 is a branched optionally substituted C1-C3 alkylene, an optionally substituted ether, an optionally substituted thioether, an optionally substituted ketone, an optionally substituted amide, polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), an optionally substituted carbocycle, an optionally substituted aryl, or an optionally substituted heteroaryl, and each R 73 are independently carboxyl or polycarboxyl, wherein the polycarboxyl contains 1 to 10, or 1 to 6, or 1 to 4 carboxyl groups, and the carboxyl groups are interconnected by alkyl, alkylene, substituted alkyl, substituted alkylene, heteroalkyl, heteroalkylene, amino, and / or amide; (c)L 70is selected from C1-C8 alkylene, C1-C8 alkylene-C(O)-, -C(O)-C1-C8 alkylene-, and -C(O)-C1-C8 alkylene-C(O)-, * is R b to the enzymatically cleavable group or to the remainder of the polar group; R 70 HA~N(R 74 -R 73 )(R 72 - R 73 ), where R 72 and R 74 are each independently selected from an optionally substituted C1-C3 alkylene, an optionally substituted ether, an optionally substituted thioether, an optionally substituted ketone, an optionally substituted amide, a polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), an optionally substituted carbocycle, an optionally substituted aryl, or an optionally substituted heteroaryl; and each R 73 are independently carboxyl or polycarboxyl, where the polycarboxyl contains 1 to 10, or 1 to 6, or 1 to 4 carboxyl groups, and the carboxyl groups are interconnected by alkyl, alkylene, substituted alkyl, substituted alkylene, heteroalkyl, heteroalkylene, amino, and / or amido.

[0130] In some embodiments, linker compounds are provided that include a polar group comprising a polymer unit and a sugar unit. In other embodiments, linker compounds are provided that include a polar group comprising at least two polymer units. In other embodiments, linker compounds are provided that include a polar group comprising a polymer unit and a carboxyl unit. In other embodiments, linker compounds are provided that include at least two polar groups. In other embodiments, linker compounds are provided that include a polar group comprising a polymer unit, a sugar unit, and a carboxyl unit. In yet other embodiments, linker compounds are provided that include a polar group comprising at least two polymer units, at least one sugar unit, and at least one carboxyl unit.

[0131] In some embodiments, linker compounds are provided that include at least one polar group attached to an enzyme-cleavable group, for example, in some embodiments, linker compounds are provided that include at least one polar group and an enzyme-cleavable group that includes at least two amino acids.

[0132] Linker Unit In some embodiments, the linker unit has the following structure (i) or (ii): TIFF2026504840000112.tif39128, During the ceremony, α- is the attachment site to the enzyme-cleavable group; β- is the site of attachment to at least one polar group; δ- is H, a site of attachment to at least one of the Drug units, or a site of attachment to a linking group attached to at least one of the Drug units; Each R a are independently H or C1-C6 alkyl; Each R b is independently halo, C 1~6 alkyl, a bond to at least one of the drug units, or a bond to at least one polar group; x is 0, 1, 2, 3, or 4; y is 0, 1, 2, or 3; R c is a bond, -C(O)-, -S(O)-, -SO2-, 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)-, -SO2-C1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, triazolyl-containing group, or a combination thereof.

[0133] In some embodiments, the Linker unit has 1 to 4 attachment sites for the Drug unit. In some embodiments, the Linker unit has 1 to 3 or 1 to 2 attachment sites for the Drug unit (D).

[0134] In some embodiments, the linker unit comprises at least one polar group, such as a polymer unit. In some embodiments, the polar group comprises at least one polymer unit and, optionally, a sugar unit and / or a carboxyl unit, or a combination thereof. In some embodiments, 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.

[0135] In some embodiments, the Linker unit is a cleavable Linker subunit. As used herein, the term "cleavable" refers to a metabolic process or reaction in an intracellular or extracellular environment that cleaves the covalent bond between the Drug unit (e.g., a cytotoxic agent) and the Linker unit, or a portion thereof, resulting in a free Drug unit or a Linker unit-Drug unit metabolite dissociated from the remainder of the Linker unit.

[0136] In some embodiments, the linker unit comprises a protease-cleavable linker subunit. In some embodiments, the linker unit is a protease-cleavable linker that is cleavable under intracellular conditions, such that cleavage of the linker unit at or within the linker unit releases the drug unit from the linker unit or from the remainder of the linker unit within the intracellular environment. For example, in some embodiments, the linker unit is cleavable by a cleaving agent present in the intracellular environment (e.g., within a lysosome, endosome, or caveolae). As used herein, the terms "cleavable under intracellular conditions," "cleaved intracellularly," and "intracellular cleavage" refer to a metabolic process or reaction within a cell that cleaves the covalent bond between the drug unit (e.g., a cytotoxic agent) and the linker unit, or a portion thereof, to produce a free drug unit within the cell or a linker unit-drug unit metabolite dissociated from the remainder of the linker unit. One advantage of using intracellular proteolytic release of the drug unit is that the activity of the drug unit is typically attenuated upon conjugation, and the serum stability of the conjugate is typically increased.

[0137] In some embodiments, the bond between the linker unit and the drug unit can be enzymatically cleaved within the linker unit by one or more enzymes, including tumor-associated proteases, to release the drug unit. The linker unit can be, for example, a peptidyl linker that is cleaved by an intracellular peptidase or protease enzyme, including but not limited to, a lysosomal or endosomal protease. Intracellular proteases or cleaving agents can include cathepsin B, C, and D and plasmin, all of which are known to hydrolyze dipeptide drug derivatives, resulting in the release of active drugs within target cells (see, for example, Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123). The peptidyl linker can be cleaved by an enzyme present in target antigen-expressing cells. For example, linker units (e.g., having Phe-Leu, Val-Ala, Val-Cit, or Gly-Phe-Leu-Gly peptides) that are cleavable by cathepsin B, a thiol-dependent protease that is highly expressed in cancer tissue, can be used.

[0138] Typically, a linker unit has at least one amino acid or at least two amino acids that form a recognition site for a protease or other cleavage agent. In certain embodiments, the linker unit has a dipeptide, tripeptide, tetrapeptide, or pentapeptide. In certain embodiments, the linker unit may contain only natural amino acids. In some embodiments, for example, the linker unit may have a Phe-Leu, Val-Ala, or Val-Cit peptide linked to a PABA group of structure (i) or (ii). Other such cleavable linker units are described, for example, in U.S. Patent No. 6,214,345, WO2004 / 010957, US20150297748, US2008 / 0166363, US20120328564, and US20200347075, each of which is incorporated herein by reference. In some embodiments, the linker unit cleavable by an intracellular protease comprises a Val-Cit peptide or a Phe-Lys peptide or a Gly-Gly-Phe-Gly linker (see, e.g., U.S. Patent Application Publication No. 2015 / 0297748).

[0139] In some embodiments, a linker unit contains one or more of the following: glycine and / or L-amino acids that form recognition and cleavage sites for proteases or other cleavage enzymes, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine.

[0140] In some embodiments, the amino acids of the linker unit have the formula shown below within square brackets: TIFF2026504840000113.tif24128, In the formula, R 190is hydrogen, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, -CH2OH, -CH(OH)CH3, -CH2CH2SCH3, -CH2CONH2, -CH2COOH, -CH2CH2CONH2, -CH2CH2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)3NH2, -(CH2)3NHCOCH3, -(CH2)3NHCHO, -(CH2)4NHC(=NH)NH2, -(CH2)4NH2, -(CH2)4NHCOCH3, -(CH2)4NHCHO, -(CH2)3NHCONH2, -(CH2)4NHCONH2, -CH2CH2CH(OH)CH2NH2, 2-pyridylmethyl-, 3-pyridylmethyl-, 4-pyridylmethyl-, phenyl, cyclohexyl, The file is TIFF2026504840000114.tif59128.

[0141] In some embodiments, the linker unit comprises one or more of the following L-(natural) amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, tryptophan, and valine. In some embodiments, the linker unit does not contain cysteine. In some embodiments, the peptidyl linker does not contain proline.

[0142] In some embodiments, the linker unit comprises one or more of the following amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, proline, tryptophan, valine, ornithine, penicillamine, β-alanine, aminoalkanoic acids, aminoalkynol acids, aminoalkanedioic acids, aminobenzoic acids, amino-heterocyclo-alkanoic acids, heterocyclo-carboxylic acids, citrulline, statins, diaminoalkanoic acids, and derivatives thereof.

[0143] In some embodiments, the linker unit comprises a protease-cleavable linker comprising a thiol-reactive maleimidocaproyl spacer or stretcher, an amino acid or peptide, and a self-immolative group. In some embodiments, the linker unit comprises a protease-cleavable linker comprising a thiol-reactive maleimidocaproyl spacer, a valine-citrulline dipeptide, and a p-amino-benzyloxycarbonyl self-immolative group.

[0144] In some embodiments, the linker unit comprises a self-stabilizing moiety comprising a maleimide group as described in WO2013 / 173337.

[0145] In some embodiments, the linker unit comprises a glucuronide cleavable moiety, see e.g., US2014 / 0031535.

[0146] In some embodiments, the following structure: TIFF2026504840000115.tif253157TIFF2026504840000116.tif252144TIFF2026504840000117.tif207150TIFF2026504840000118.tif207154TIFF2026504840000119.tif68130, In the formula, R C is a bond or C 1~6 is alkylene; the wavy line on the amino group indicates the attachment site for a Stretcher group or indicates an H before attachment to the Stretcher group; β- is the attachment site to at least one polar group; the benzyl H on the benzyl OH may be replaced with a bond to at least one of the Drug units or a bond to a linking group attached to at least one of the Drug units.

[0147] In some embodiments, at least one of the drug units is directly attached to the benzyl-O at δ. In some embodiments, at least one of the drug units is indirectly attached via a linking group. The linking group may be any suitable group for attaching at least one drug unit to the benzyl-O— that allows for release of the active drug unit or an active derivative of the linking group-drug unit. In some embodiments, the linking group is —NH—CH—CH—CH—C(O)—, the drug unit is exatecan, and the released drug unit is DXd. (See, e.g., U.S. Patent Application Publication No. 2019 / 000898.) In other embodiments, the linking group is —C(O)—NH—CH—CH—CH—C(O)—.

[0148] In some embodiments, a linker compound is provided, wherein the linker unit is a cleavable linker unit.For example, in some embodiments, the enzyme-cleavable group comprises a peptide that can be cleaved by an intracellular protease.In some embodiments, the intracellular protease is cathepsin B.In more specific embodiments, the cleavable peptide comprises a valine-citrulline peptide, a valine-alanine peptide, a valine-lysine peptide, a phenylalanine-lysine peptide, or a glycine-glycine-phenylalanine-glycine peptide.

[0149] In some embodiments, the following structure: TIFF2026504840000120.tif54128TIFF2026504840000121.tif219137TIFF2026504840000122.tif244112TIFF20 26504840000123.tif220115TIFF2026504840000124.tif207117TIFF2026504840000125.tif241118TIFF2026504 840000126.tif243111TIFF2026504840000127.tif192135TIFF2026504840000128.tif203115TIFF2026504840000129.tif195120TIFF2026504840000130.tif198112TIFF2026504840000131.tif205155, In the formula, R c is a bond or C 1~6 is alkylene; the wavy line on the amino group indicates a bond site for a Stretcher group or indicates an H before attachment to a Stretcher group; β- is a bond site to at least one polar group; the H on the benzyl OH may be replaced with a bond to at least one of the Drug units or a bond site to at least one of the Drug units.

[0150] In some embodiments, the following structure: TIFF2026504840000132.tif235158TIFF2026504840000133.tif205158TIFF2026504840000134.tif19415 7TIFF2026504840000135.tif209158TIFF2026504840000136.tif228157TIFF2026504840000137.tif2301 57TIFF2026504840000138.tif226157TIFF2026504840000139.tif211157TIFF2026504840000140.tif172 158TIFF2026504840000141.tif206158TIFF2026504840000142.tif231158TIFF2026504840000143.tif23 5158TIFF2026504840000144.tif216157TIFF2026504840000145.tif250158TIFF2026504840000146.tif2 32158TIFF2026504840000147.tif247158TIFF2026504840000148.tif225158TIFF2026504840000149.tif 155157TIFF2026504840000150.tif221158TIFF2026504840000151.tif205158TIFF2026504840000152.tif250157TIFF2026504840000153.tif203162TIFF2026504840000154.tif54140, wherein the wavy line on the amino group indicates the site of attachment to the Stretcher group; alternatively, it represents an H prior to attachment to the Stretcher group, and the H on the benzyl OH may be replaced with a bond to at least one of the Drug units or a bond to a linking group attached to at least one of the Drug units.

[0151] In some aspects, there is provided a linker compound as described above, wherein the linker unit is attached to the side chain of the enzyme-cleavable group subunit.

[0152] In some embodiments, the linker compounds described above are provided, wherein the enzyme-cleavable group is attached to the Stretcher group by a non-peptide linking group. For example, in some embodiments, the non-peptide linking group is an optionally substituted C1-C 10 Alkylene, optionally substituted C2-C 10 Alkenylene, optionally substituted C2-C 10 alkynylene, or optionally substituted polyethylene glycol.

[0153] In some aspects, there is provided a linker compound as described above that includes a stretcher group attached to an enzyme-cleavable group.

[0154] Stretcher base In some embodiments, the following: a linker compound is provided that includes a stretcher group selected from TIFF2026504840000155.tif91151; In the formula, 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 to 26), -C1-C8 alkylene-(CH2-O-CH2) b -(wherein b is 1 to 26), -C1 to C8 alkylene-(CH2-O-CH2) b -C1-C8 alkylene- (where b is 1 to 26), -arylene-, -C1-C 10 Alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1~C 10 Alkylene-(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~C10 Alkylene -C(=O)-, -C1 to C 10 Alkylene-C(O)NH-C1-C8 alkylene-[O-CH2-CH2] n -C(O)- (where n is 1 to 26), C1 to C 10 Heteroalkylene-C(=O)-, -C1-C8 alkylene-(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 to 26), -C1-C8 alkylene-(CH2-O-CH2) b -C1-C8 alkylene-C(=O)- (wherein b is 1 to 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 to 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 to C 10 Alkylene-NH-, -C1~C 10 Heteroalkylene-NH-, -C1-C8 alkylene-(CH2-O-CH2) b -NH- (wherein b is 1 to 26), -(CH2-O-CH2) b -C1-C8 alkylene-NH- (where b is 1 to 26), -C1-C8 alkylene-(CH2-O-CH2) b -C1-C8 alkylene-NH- (where b is 1 to 26), -C1-C8 alkylene-(C(=O))-NH-(CH2-O-CH2) b-C(=O)- (wherein b is 1 to 26), -C1 to C8 alkylene-(C(=O))-NH-(CH2-O-CH2) b -C1-C8 alkylene-C(=O)- (wherein b is 1 to 26), -C1-C8 alkylene-NH-(C(=O))-(CH2-O-CH2) b -NH- (wherein b is 1 to 26), -C1 to C8 alkylene-NH-(C(=O))-(CH2-O-CH2) b -C1-C8 alkylene-NH- (where b is 1 to 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-arylene-S-, -arylene-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 10 Alkylene-(C3-C8 heterocyclo)-S- or -(C3-C8 heterocyclo)-C1-C 10 alkylene-S-; or wherein the Stretcher group is maleimide (C1-C 10 Alkylene-C(O)-, maleimide (CH2OCH2) p2 (C1~C 10Alkylene)C(O)-, Maleimide (C1-C 10 alkylene) (CH2OCH2) p2 C(O)-, or open ring forms thereof, where p2 is 1-26; where * is the site of attachment to the targeting group and the wavy line is the site of attachment to the enzyme-cleavable group.

[0155] In some embodiments, the following: a linker compound comprising a stretcher group selected from TIFF2026504840000156.tif211153; In the formula, wavy line TIFF2026504840000157.tif2128 shows the attachment site of the stretcher group to the enzyme-cleavable group, and the attachment site to the targeting group is on the maleimide, primary amine, or alkyne functionality.

[0156] Linker Compounds In some embodiments, the following structure: TIFF2026504840000158.tif250158TIFF2026504840000159.tif242158TIFF2026504840000160.t if229158TIFF2026504840000161.tif253159TIFF2026504840000162.tif202158TIFF2026504840 000163.tif241157TIFF2026504840000164.tif227158TIFF2026504840000165.tif243158TIFF20 26504840000166.tif231158TIFF2026504840000167.tif241158TIFF2026504840000168.tif2551 58TIFF2026504840000169.tif253157TIFF2026504840000170.tif214158TIFF2026504840000171 .tif248158TIFF2026504840000172.tif244158TIFF2026504840000173.tif204158TIFF20265048 40000174.tif235158TIFF2026504840000175.tif210158TIFF2026504840000176.tif224158TIFF2026504840000177.tif217166TIFF2026504840000178.tif137164; wherein the H on the benzyl OH may be replaced with a bond to at least one Drug unit or a bond to a linking group attached to at least one Drug unit.

[0157] In some embodiments, a Drug-Linker Compound is provided that comprises a linker compound described herein attached at a binding site to at least one Drug Unit or to a linking group attached to at least one Drug Unit.

[0158] Drug Unit In some embodiments, the linker is attached to at least one drug unit. As used herein, the term "drug unit" or drug refers to a cytotoxic agent (such as a chemotherapeutic agent or drug), an immunomodulatory agent, a nucleic acid (including siRNA), a growth inhibitory agent, a toxin (e.g., a protein toxin, an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or a fragment thereof), a radioisotope, a PROTAC, and other compounds that become active against a target cell when delivered to the target cell.

[0159] In some embodiments, a Drug-Linker is provided wherein the Drug unit is selected from a cytotoxic agent, an immunomodulator, a nucleic acid, a growth inhibitor, a PROTAC, a toxin, a radioisotope, and a chelating ligand.

[0160] cytotoxic substances In some embodiments, the drug unit is a cytotoxic agent. "Cytotoxic agent" refers to an agent that has a cytotoxic effect on a cell. "Cytotoxic effect" refers to the depletion, elimination, and / or death of the target cell. Cytotoxic agents include, for example, tubulin-disrupting agents, topoisomerase inhibitors, DNA minor groove binders, and DNA alkylating agents.

[0161] In some embodiments, the cytotoxic agent is selected from the group consisting of auristatin, maytansinoid, camptothecin, duocarmycin, and calicheamicin. In some embodiments, the cytotoxic agent is an auristatin. In other embodiments, the cytotoxic agent is MMAE or MMAF. In some embodiments, the cytotoxic agent is camptothecin. In some embodiments, the cytotoxic agent is exatecan, SN-38, or DxD. In certain embodiments, the cytotoxic agent is RS-exatecan or SS-exatecan. In other embodiments, the cytotoxic agent is calicheamicin. In some embodiments, the cytotoxic agent is a maytansinoid. In certain embodiments, the maytansinoid is maytansine, maytansinol, or ansamatocin-2.

[0162] Tubulin disrupting agents include, for example, auristatins, dolastatins, tubulysins, colchicine, vinca alkaloids, taxanes, cryptophycins, maytansinoids, hemiasterins, and other tubulin disrupting agents. Auristatins are derivatives of the natural product dolastatin 10. Examples of auristatins include MMAE (N-methylvaline-valine-dolaisoleuine-dolaproine-norephedrine), MMAF (N-methylvaline-valine-dolaisoleuine-dolaproine-phenylalanine), and AFP (see WO2004 / 010957 and WO2007 / 008603). Other auristatin-like compounds are disclosed, for example, in U.S. Patent Application Publication Nos. US2021 / 0008099, US2017 / 0121282, US2013 / 0309192, and US2013 / 0157960. Dolastatins include, for example, dolastatin 10 and dolastatin 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 No. US2001 / 0018422). Additional dolastatin derivatives contemplated for use herein are disclosed in U.S. Patent No. 9,345,785, which is incorporated herein by reference.

[0163] Tubulysins include, but are not limited to, tubulysin D, tubulysin M, tubuphenylalanine, and tubutyrosine. WO2017 / 096311 and WO / 2016-040684 describe tubulysin analogs, including tubulysin M.

[0164] Colchicine includes, but is not limited to, colchicine and CA-4.

[0165] Vinca alkaloids include, but are not limited to, vinblastine (VBL), vinorelbine (VRL), vincristine (VCR), and vindesine (VOS).

[0166] Taxanes include, but are not limited to, paclitaxel and docetaxel.

[0167] Cryptophycins include, but are not limited to, cryptophycin-1 and cryptophycin-52.

[0168] Maytansinoids include, but are not limited to, maytansine, maytansinol, maytansine analogs in DM1, DM3, and DM4, and ansamatocin-2. Exemplary maytansinoid drug moieties include those with modified aromatic rings, such as C-19-deschloro (U.S. Pat. No. 4,256,746) (prepared by lithium aluminum hydride reduction of ansamitocin P2); C-20-hydroxy (or C-20-desmethyl) + / -C-19-deschloro (U.S. Pat. Nos. 4,361,650 and 4,307,016) (prepared by demethylation with Streptomyces or Actinomyces, or dechlorination with LAH); and C-20-desmethoxy, C-20-acyloxy (-OCOR), + / -deschloro (U.S. Pat. No. 4,294,757) (prepared by acylation with acyl chloride), as well as those with modifications at other positions.

[0169] Maytansinoid drug moieties may have modifications, such as C-9-SH (U.S. Pat. No. 4,424,219) (prepared by reaction of maytansinol with H2S or P2S5); C-14-alkoxymethyl (demethoxy / CH2OR) (see U.S. Pat. No. 4,331,598); C-14-hydroxymethyl or acyloxymethyl (CH2OH or CHOAc) (see U.S. Pat. No. 4,450,254) (prepared from Nocardia sp.); C-15-hydroxy / acyloxy (see U.S. Pat. No. 4,364,866) (prepared by conversion of maytansinol by Streptomyces); C-15-methoxy (see U.S. Pat. Nos. 4,313,946 and 4,315,929) (prepared from Trewia nudiflora nudiflora); C-18-N-demethyl (see U.S. Pat. Nos. 4,362,663 and 4,322,348) (prepared by demethylation of maytansinol with Streptomyces); and 4,5-deoxy (see U.S. Pat. No. 4,371,533) (prepared by titanium trichloride / LAH reduction of maytansinol).

[0170] Hemiasterins include, but are not limited to, hemiasterin and HTL-286.

[0171] Other tubulin disrupting agents include taccalonolide A, taccalonolide B, taccalonolide AF, taccalonolide AJ, taccalonolide Al-epoxide, discodermolide, epothilone A, epothilone B, and laulimalide.

[0172] In some embodiments, the cytotoxic agent may be a topoisomerase inhibitor, such as camptothecin. Exemplary camptothecins include, for example, camptothecin, irinotecan (also known as CPT-11), belotecan, (7-(2-(N-isopropylamino)ethyl)camptothecin), topotecan, 10-hydroxy-CPT, SN-38, exatecan, and the exatecan analog DXd (see US20150297748). In some embodiments, the cytotoxic agent is exatecan, SN-38, or DxD. In some embodiments, the cytotoxic agent is RS exatecan or SS exatecan. Other camptothecins are disclosed in WO1996 / 021666, WO00 / 08033, US2016 / 0229862, and WO2020 / 156189.

[0173] In some embodiments, the cytotoxic agent is a duocarmcycin, including the synthetic analogs KW-2189 and CBI-TMI.

[0174] immunomodulator In some embodiments, the drug unit is an immunomodulator. The immunomodulator may be, for example, a TLR7 and / or TLR8 agonist, a STING agonist, a RIG-I agonist, or other immunomodulator. In some embodiments, the immunomodulator is selected from a TRL7 agonist, a TLR8 agonist, a STING agonist, or a RIG-I agonist.

[0175] In some embodiments, the immunomodulator is a TLR7 agonist. In certain embodiments, the TLR7 agonist is imidazoquinoline, imidazoquinoline amine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzimidazol-2-amine, tetrahydropyridopyrimidine, heteroaromatic azido-2,2-dioxide, benzonaphthyridine, guanosine analog, adenosine analog, thymidine homopolymer, ssRNA, CpG-A, polyG10, or polyG3. In some embodiments, the immunomodulator is a TLR8 agonist. In certain embodiments, 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-benzimidazol-2-amine, tetrahydropyridopyrimidine or ssRNA. In some embodiments, the immunomodulator is a STING agonist. In another embodiment, the immunomodulator is a RIG-I agonist. In certain embodiments, the RIG-I agonist is selected from KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400 and KIN2000.

[0176] In some embodiments, the Drug Unit is an immunomodulator, such as a TLR7 and / or TLR8 agonist. In some embodiments, the TLR7 agonist is selected from imidazoquinolines, imidazoquinoline amines, thiazoquinolines, aminoquinolines, aminoquinazolines, pyrido[3,2-d]pyrimidine-2,4-diamines, pyrimidine-2,4-diamines, 2-aminoimidazoles, 1-alkyl-1H-benzimidazol-2-amines, tetrahydropyridopyrimidines, heteroaromatic azides-2,2-dioxides, benzonaphthyridines, guanosine analogs, adenosine analogs, thymidine homopolymers, ssRNA, CpG-A, polyG10, and polyG3. In some embodiments, the TLR7 agonist is selected from imidazoquinolines, imidazoquinoline amines, thiazoquinolines, aminoquinolines, aminoquinazolines, pyrido[3,2-d]pyrimidine-2,4-diamines, pyrimidine-2,4-diamines, 2-aminoimidazoles, 1-alkyl-1H-benzimidazol-2-amines, tetrahydropyridopyrimidines, heteroaromatic azides-2,2-dioxides, or benzonaphthyridines. In some embodiments, the TLR7 agonist is a non-naturally occurring compound. Examples of TLR7 modulators include GS-9620, GSK-2245035, imiquimod, resiquimod, 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 compounds disclosed in US20160168164, US20150299194, US20110098248, US20100143301, and US20090047249.

[0177] In some embodiments, the TLR8 agonist is selected from benzazepines, imidazoquinolines, thiazoloquinolines, aminoquinolines, aminoquinazolines, pyrido[3,2-d]pyrimidine-2,4-diamines, pyrimidine-2,4-diamines, 2-aminoimidazoles, 1-alkyl-1H-benzimidazol-2-amines, tetrahydropyridopyrimidines, or ssRNA. In some embodiments, the TLR8 agonist is selected from benzazepines, imidazoquinolines, thiazoloquinolines, aminoquinolines, aminoquinazolines, pyrido[3,2-d]pyrimidine-2,4-diamines, pyrimidine-2,4-diamines, 2-aminoimidazoles, 1-alkyl-1H-benzimidazol-2-amines, and tetrahydropyridopyrimidines. In some embodiments, the TLR8 agonist is a non-natural compound. Examples of TLR8 agonists include motolimod, resiquimod, 3M-051, 3M-052, MCT-465, IMO-4200, VTX-763, and VTX-1463.

[0178] In some embodiments, the TLR8 agonist can be any of the compounds described in WO2018 / 170179, WO2020 / 056198, and WO2020056194.

[0179] Other TLR7 and TLR8 agonists are described, for example, in WO2016142250, WO2017046112, WO2007024612, WO2011022508, WO2011022509, WO2012045090, WO2012097173, WO2012097177, WO2017079283, US20160008374, US20160194350, US20160289229, U.S. Patent No. 6,043,238, US20180086755, WO2017216054, WO2017190669, WO2017202704, WO2017202703, WO20170071944, US2014 0045849, US20140073642, WO2014056953, WO2014076221, WO2014128189, US2014035 0031, WO2014023813, US20080234251, US20080306050, US20100029585, US201100924 85, US20110118235, US20120082658, US20120219615, US20140066432, US20140088085, US20140275167, and US20130251673, WO2018198091, and US20170131421.

[0180] In some embodiments, the immunomodulator is a STING agonist. Examples of STING agonists include those disclosed in, for example, WO2020059895, WO2015077354, WO2020227159, WO2020075790, WO2018200812, and WO2020074004.

[0181] In some embodiments, the immunomodulator is a RIG-I agonist. Examples of RIG-I agonists include KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400, and KIN2000.

[0182] toxin In some embodiments, the Drug Unit is an enzymatically active toxin or fragment thereof, including, but not limited to, diphtheria A chain, non-binding active fragment of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alphasarcin, Aleurites fordii protein, dianthin protein, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogenin, restrictocin, phenomycin, enomycin, and a trichothecene.

[0183] radioactive isotope In some embodiments, the Drug Unit is a radioactive atom. A variety of radioisotopes are available for generating radioconjugates. Examples include 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.

[0184] PROTAC In some embodiments, the drug unit is a proteolysis-directed chimera (PROTAC). PROTACs are described, for example, in U.S. Patent Application Publication Nos. 20210015942, 20210015929, 20200392131, 20200216507, US20200199247, and US20190175612, the disclosures of which are incorporated herein by reference.

[0185] Ligand In some embodiments, the Drug Unit comprises a ligand that can be bound by a carboxyl unit, such as platinum (Pt), ruthenium (Ru), rhodium (Rh), gold (Au), silver (Ag), copper (Cu), molybdenum (Mo), titanium (Ti), or iridium (Ir); a radioisotope, such as 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.

[0186] Drug-Linker Compounds In some embodiments, a Drug-Linker Compound is provided that comprises a linker compound described above having at least one Drug unit attached via a linking site.

[0187] In some embodiments, provided are Drug-Linker Compounds comprising a Drug unit selected from a cytotoxic agent, an immunomodulator, a nucleic acid, a growth inhibitor, a PROTAC, a toxin, a radioisotope, and a chelating ligand. In more specific embodiments, the Drug unit is a cytotoxic agent. In more specific embodiments, the cytotoxic agent is selected from the group consisting of an auristatin, a maytansinoid, a camptothecin, a duocarmycin, and a calicheamicin. In some embodiments, the cytotoxic agent is an auristatin. In some embodiments, the cytotoxic agent is MMAE or MMAF. In some embodiments, the cytotoxic agent is SS-exatecan. In some embodiments, the cytotoxic agent is RS-exatecan. In some embodiments, the cytotoxic agent is camptothecin. In some embodiments, the cytotoxic agent is exatecan or SN-38. In some embodiments, the cytotoxic agent is exatecan. In some embodiments, the cytotoxic agent is DxD. In some embodiments, the cytotoxic agent is calicheamicin. In some embodiments, the cytotoxic agent is a maytansinoid. In more specific embodiments, the maytansinoid is maytansine, maytansinol, or ansamatocin-2.

[0188] In some embodiments, a drug-linker compound is provided, comprising the linker compound described above, wherein a drug unit is linked via a binding site, wherein the drug unit is an immunomodulator.For example, in some embodiments, the immunomodulator is selected from a TLR7 agonist, a TLR8 agonist, a STING agonist, or a RIG-I agonist.In more specific embodiments, the immunomodulator is a TLR7 agonist. In more specific embodiments, the TLR7 agonist is an imidazoquinoline, imidazoquinoline amine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzimidazol-2-amine, tetrahydropyridopyrimidine, heteroaromatic thiol azide-2,2-dioxide, benzonaphthyridine, guanosine analog, adenosine analog, thymidine homopolymer, ssRNA, CpG-A, polyG10, or polyG3. In certain embodiments, the immunomodulator is a TLR8 agonist. In more specific embodiments, 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-benzimidazol-2-amine, tetrahydropyridopyrimidine or ssRNA. In some embodiments, the immunomodulator is a STING agonist. In another embodiment, the immunomodulator is a RIG-I agonist. For example, in some embodiments, the RIG-I agonist is selected from KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400 and KIN2000.

[0189] In some embodiments, a drug-linker compound is provided, comprising the linker compound described above, to which a drug unit is attached via a linking moiety, wherein the drug unit is a chelating ligand. For example, in some embodiments, the chelating ligand is selected from platinum (Pt), ruthenium (Ru), rhodium (Rh), gold (Au), silver (Ag), copper (Cu), molybdenum (Mo), titanium (Ti), or iridium (Ir); a radioisotope, such as 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.

[0190] In some embodiments, the compound of formula (A): A drug linker compound represented by the structure TIFF2026504840000179.tif39128 or a salt thereof is provided, During the ceremony, (i) β is R 20 (-R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 ) z where: R 20 is NH, TIFF2026504840000180.tif38128; R 21 and R 22 each independently represents a bond or C1-C3 alkylene; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; and a —C(O)-polyhydroxyl group; provided that R 24 and R 25 cannot both be H; z is 1 or 2; n20 is 2-26; (ii)R ais H or C1-C6 alkyl; (iii)R c is a bond, -C(O)-, -S(O)-, -SO2-, C 1~6 Alkylene, C 1~6 Alkynylene, or C 1~6 alkynylene-triazolyl; (iv)R 1 is a bond, -C(O)-, or C 1~6 is alkylene; (v) δ is selected from drugs; (vi) α is Represented by TIFF2026504840000181.tif35128; where R 2 is a peptide having 2 to 5 amino acids; R 3 is -C1~C 10 Alkylene -C(=O)-, -C1 to C 10 Alkylene-, -C1~C 10 Alkylene-C(O)NH-C1-C8 alkylene-[O-CH2-CH2] n -C(O)- (where n is 1 to 26), or -C1 to C8 alkylene-(CH2-O-CH2) b -C(=O)- (wherein b is 1 to 26).

[0191] In some embodiments, for the drug-linker, R 20 teeth The file is TIFF2026504840000182.tif27128.

[0192] In some embodiments, z is 1 for the drug-linker.

[0193] In some embodiments, z is 2 for the drug-linker.

[0194] In some embodiments, for the drug-linker, R 24 and R 25 are each independently selected from polyhydroxyl groups.

[0195] In some embodiments, for the drug-linker, R 21 is C1-C3 alkylene. In some cases, R 21 is a C2 alkylene.

[0196] In some embodiments, for the drug-linker, R 22 is a bond.

[0197] In some embodiments, for the drug-linker, β is TIFF2026504840000183.tif83128, and n20 is 4 to 12.

[0198] In some embodiments, for the drug-linker, β is The file is TIFF2026504840000184.tif59128.

[0199] In some embodiments, for the drug-linker, R 2 teeth Selected from TIFF2026504840000185.tif58129.

[0200] In some embodiments, for the drug-linker, R 2 teeth Selected from TIFF2026504840000186.tif57128.

[0201] In some embodiments, for the drug-linker, R 3 teeth Selected from TIFF2026504840000187.tif16128.

[0202] In some embodiments, for the drug-linker, R 3 teeth Selected from TIFF2026504840000188.tif15128.

[0203] In some embodiments, for the drug-linker, R cis -C(O)- or C 1~6 It is an alkynylene-triazolyl.

[0204] In some embodiments, for the drug-linker, R c is -C(O)-.

[0205] In some embodiments, for the drug-linker, R 1 is -C(O)-.

[0206] In some embodiments, for the drug-linker, the drug is selected from cytotoxic agents. In some embodiments, the drug is MMAE or MMAF. In some embodiments, the drug is MMAE. In some embodiments, the drug is exatecan.

[0207] In some embodiments, for the drug-linker, the drug is selected from an immunomodulator.

[0208] In some embodiments, for the drug-linker, δ is TIFF2026504840000189.tif68151. In some embodiments, δ is TIFF2026504840000190.tif24128. In some embodiments, δ is The file is TIFF2026504840000191.tif31128.

[0209] In some embodiments, the wavy bond indicates the point of attachment to the drug-linker.

[0210] In some embodiments, the following structure: TIFF2026504840000192.tif130158TIFF2026504840000193.tif226160TIFF2026504840000194.tif218161TIF F2026504840000195.tif253158TIFF2026504840000196.tif255157TIFF2026504840000197.tif254157TIFF20 26504840000198.tif251159TIFF2026504840000199.tif246158TIFF2026504840000200.tif207159TIFF20265 04840000201.tif186158TIFF2026504840000202.tif196162TIFF2026504840000203.tif210158TIFF20265048 40000204.tif228162TIFF2026504840000205.tif192158TIFF2026504840000206.tif213158TIFF20265048400 00207.tif247158TIFF2026504840000208.tif233158TIFF2026504840000209.tif245158TIFF20265048400002 Drug-linker compounds selected from: 10.tif253157TIFF2026504840000211.tif232158TIFF2026504840000212.tif220158TIFF2026504840000213.tif252164TIFF2026504840000214.tif235164TIFF2026504840000215.tif138164 are provided.

[0211] In some embodiments, the Drug-Linker may be a stereoisomer of one of the above structures.

[0212] Targeting Group In some embodiments, the linker is attached to the targeting group (also referred to herein as "targeting unit") to form a targeting unit-linker. In some embodiments, the linker is attached to the targeting group via the Stretcher unit and to the Drug unit via binding site δ to form a conjugate. In some embodiments, the targeting group is a protein, polypeptide, or peptide. The targeting group may be an antibody, an antigen-binding portion thereof, or a non-antibody targeting group. A non-antibody targeting group may also be referred to as a non-antibody scaffold.

[0213] In some embodiments, the targeting group specifically binds to a target molecule. As used herein, "specifically binds" refers to a targeting group (e.g., an antibody or portion thereof) described herein specifically binds to a target molecule at a KD 10 -5 M (10000 nM) or less, e.g., 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M or less refers to the ability to bind to a target.Specific binding can be affected, for example, by the affinity and avidity of the targeting group and the concentration of the target polypeptide.Those skilled in the art can use any suitable method, such as titrating the binding substance in a suitable cell binding assay, to determine the appropriate conditions under which the antibody, antibody binding moiety and non-antibody scaffold described herein selectively bind to the target.The targeting group that specifically binds to the target is not displaced by a non-similar competitor.In certain embodiments, a targeting unit is said to specifically bind to its target when it preferentially recognizes its target in a complex mixture of proteins and / or macromolecules.

[0214] As used herein, the term "antibody" refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen-binding site that specifically binds to a target antigen. The term generally refers to antibodies that contain two immunoglobulin heavy chain variable regions and two immunoglobulin light chain variable regions, and includes full-length antibodies (having heavy and light chain constant regions).

[0215] Each heavy chain typically consists of a variable region (abbreviated as VH region) 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 region) and a constant region. The light chain constant region is a CL domain. The VH and VL regions may be further divided into highly variable regions called complementarity-determining regions (CDRs) and interspersed conserved regions called framework regions (FRs). Each VH and VL region thus contains three CDRs and four FRs, arranged from the N-terminus to the C-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. This structure is well known to those skilled in the art.

[0216] As used herein, the term "antigen-binding portion" of an antibody refers to a portion of an antibody that contains the VH and / or VL sequences or CDRs of the antibody and specifically binds to a target antigen. Examples of antigen-binding portions include Fab, Fab', F(ab')2, Fv, scFv, disulfide-linked Fv, single domain antibodies (also known as VHH, VNAR, sdAb, or nanobodies), 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 the following: (i) Fab is a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab')2 is a bivalent fragment comprising two Fab fragments linked together at the hinge region via disulfide bridges; and (iii) Fv is composed of the VL and VH domains. The two domains of the Fv fragment, i.e., VL and VH, are encoded by separate coding regions but may be linked together by a synthetic linker, such as a poly-G4S amino acid sequence (disclosed as SEQ ID NO:1, "(G4S)" where n=1 to 5). n"), allowing them to be prepared as a single protein chain in which the VL and VH regions together form a monovalent molecule (known as a single-chain Fv or scFv). 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 "diabodies," are also included herein. Diabodies are bivalent, bispecific antibodies in which the VH and VL regions are expressed on a single polypeptide chain, but the linker connecting the VH and VL regions is too short to allow the two regions to bind on the same chain, thereby forcing the VH and VL regions to pair with complementary regions (VL and VH, respectively) of different chains to form two antigen-binding sites (see, e.g., Holliger, R, et al. (1993) Proc. Natl. Acad. Sci. USA 90:64446448; Poljak, R. J, et al. (1994) Structure 2:1121-1123).

[0217] A single domain antibody is an antigen-binding portion of an antibody that contains a single monomeric variable antibody region. Single domain antibodies can be derived from the variable region of an antibody heavy chain from a camelid (e.g., a nanobody or VHH portion). Furthermore, the term single domain antibody includes autonomous human heavy chain variable domains (aVH) or VNAR portions derived from sharks (see, e.g., Hasler et al., Mol. Immunol. 75:28-37, 2016).

[0218] Techniques for producing single domain antibodies (e.g., 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). VHHs can have strong antigen-binding capabilities and can interact with novel epitopes that are inaccessible to conventional VH-VL pairs (see, e.g., Muyldermans et al., 2001). Alpaca serum IgG contains approximately 50% heavy chain-only IgG antibodies (HCAbs) of camelid origin (see, e.g., Maass et al., 2007). Alpacas can be immunized with antigens, and VHHs that bind to and neutralize the target antigen 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 be used to isolate antibody fragments using standard biopanning techniques well known in the art (see, e.g., Maass et al., 2007).

[0219] In some embodiments, the targeting group is an antibody or antigen-binding portion thereof, and is 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, the IgG-scFv is an IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, svFc-(L)IgG, 2scFV-IgG, or IgG-2scFv. See, for example, 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.

[0220] In some embodiments, the targeting group is a cancer-associated antigen, e.g., CD19, CD20, CD30, CD33, CD38, CA125, HER2, MUC-1, prostate-specific membrane antigen (PSMA), CD44 surface adhesion molecule, mesothelin (MLSN), carcinoembryonic antigen (CEA), epidermal growth factor receptor (EGFR), EGFRvIII, vascular endothelial growth factor receptor 2 (VEGFR2), high molecular weight melanoma-associated antigen (HMW-MAA), MAGE-A1, IL-13 R-a2, GD2, 1p19q, ABL1, AKT1, ALK, APC, AR, ATM, BRAF, BRCA1, BRCA2, cKIT, cMET, CSF1R, CTNNB1, FGFR1, FGFR2, FLT3, GNA11, G NAQ, GNAS, HRAS, IDH1, IDH2, JAK2, KDR(VEGFR2), KRAS, MGMT, MGMT-Me, MLH1, MPL, NOTCH1, NRAS, PDGFRA, Pgp, PIK3CA, PR, PTE N, RET, RRM1, SMO, SPARC, TLE3, TOP2A, TOPO1, TP53, TS, TUBB3, VHL, CDH1, ERBB4, FBXW7, HNF1A, JAK3, NPM1, PTPN11, RB1, SMA D4, SMARCB1, STK1, MLH1, MSH2, MSH6, PMS2, ROS1, ERCC1, 5T4(TPBG), B7-H3, CCR7, CD105, CD22, CD46, CD47, CD56, CD70, CD71, It specifically binds to CD79b, CDH6, CLDN6, CLDN18.2, CLEC12A, DLL3, DR5, ERBB3 (HER3), EPCAM, FOLR1, IGF1R, IL2RA (CD25), IL3RA, ITGB6, LIV-1, LRRC15, mesothelin (MSLN), NaPi2b (SLC34A2), Nectin-4, PTK7, ROR1, SEZ6, SLC44A4, SLITRK6, tissue factor (TF), TROP2, or B7-H4. According to the invention, the terms "cancer-associated antigen," "tumor antigen," "tumor-expressed antigen," "cancer antigen," "cancer-associated antigen," and "cancer-expressed antigen" are synonymous and are used interchangeably herein.

[0221] In some embodiments, the targeting group specifically binds to a target such as CD19, CD20, CD30, CD33, CD70, LIV-1, or EGFRv3.

[0222] In some embodiments, the targeting group is a target-binding antibody (or fragment thereof) having a sequence disclosed in US 2022 / 0048951 to Leuschner et al. and / or US 2022 / 0016258 to Lerchen et al. Non-limiting examples of monoclonal antibodies include rituximab (Rituxan®), trastuzumab (Herceptin®), pertuzumab (Perjeta®), bevacizumab (Avastin®), ranibizumab (Lucentis®), cetuximab (Erbitux®), alemtuzumab (Campus®), panitumumab ( Vectibix (registered trademark), ibritumomab (Zevalin (registered trademark), tositumomab (Vexar (registered trademark), ipilimumab, zalutumumab, dalotuzumab, figitumumab, ramucirumab, galiximab, faretuzumab, ocrelizumab, ofatumumab (Arzera (registered trademark), CD20 antibody 2F2 (HuMax-CD20), 7D8, IgM2C6, IgG1 Antibodies that can be used in combination with the conjugates of the invention include anti-LHRH receptor antibodies such as 2C6, 11B8, B1, 2H7, LT20, 1FS, or AT80 (see Teeling et al., J. Immunol. 177:362-371 (2006)), daclizumab (Zenapax®), and clones A9E4, F1G4, AT2G7, GNRH03, GNRHR2, among others.

[0223] In some embodiments, the targeting group is a non-antibody scaffold. Such non-antibody scaffolds include, for example, affibodies, affilins, anticalins, atrimers, avimers, bicyclic peptides, cysteine ​​knots, DARPins, FN3 scaffolds (e.g., adnectins, centilins, pronectins, and Tn3), finomers, Kunitz domains, and O-bodies. (See, for example, Vazquez-Lombardi et al., Drug Discovery Today 20(10):1271 (2015) and the references cited therein.) Such non-antibody proteins include, for example, affibodies, affilins, anticalins, atrimers, avimers, bicyclic peptides, cysteine ​​knots, DARPins, FN3 scaffolds (e.g., adnectins, centilins, pronectins, and Tn3), finomers, Kunitz domains, and O-bodies. (See, e.g., Vazquez-Lombardi et al., Drug Discovery Today 20(10):1271 (2015) and references cited therein.) Non-antibody scaffolds can be considered to fall into two structural categories: domain-sized constructs (in the 6-20 kDa range) and constrained peptides (in the 2-4 kDa range). Domain-sized non-antibody scaffolds include, but are not limited to, affibodies, affilins, anticalins, atrimers, DARPins, FN3 scaffolds (such as adnectins and centrins), finomers, Kunitz domains, pronectins, and O-bodies. Peptide-sized non-antibody scaffolds include, for example, avimers, bicyclic peptides, and cysteine ​​knots. Non-antibody protein scaffolds can be considered to fall into two structural categories: domain-sized constructs (in the 6-20 kDa range) and constrained peptides (in the 2-4 kDa range). Domain-sized non-antibody scaffolds include, but are not limited to, affibodies, affilins, anticalins, atrimers, DARPins, FN3 scaffolds (such as adnectins and centirins), fynomers, Kunitz domains, pronectins, and O-bodies.Peptide-sized non-antibody scaffolds include, for example, avimers, bicyclic peptides and cysteine ​​knots.These non-antibody scaffolds and the proteins or peptides that they are based on or derived from are discussed, for example, by Simeon and Chen, Protein Cell 9(1): 3-14 (2018); Vazquez-Lombardi et al., Drug Discovery Today 20: 1271-1283 (2015) and Binz et al., Nature Biotechnol. 23: 1257-1268 (2005), the contents of each of which are incorporated herein by reference in their entirety.

[0224] The advantages of using non-antibody scaffolds include improved affinity, target neutralization, and stability. Various non-antibody scaffolds can also overcome some of the limitations of antibody scaffolds, for example, in terms of tissue penetration, compactness, and thermal stability. Some non-antibody scaffolds can be more easily constructed when bispecific constructs are desired, for example, without being hindered by the association of light chains. Methods for constructing constructs on non-antibody scaffolds are known to those skilled in the art.

[0225] Thus, in some embodiments, the targeting group may comprise a non-antibody scaffold.Thus, in some embodiments, the targeting group may comprise a non-antibody scaffold protein. Those skilled in the art will recognize that in some embodiments, the targeting group may be selected from a wide variety of scaffolds, including, for example, an adnectin scaffold or portion thereof derived from human 10th fibronectin type III domain (10Fn3); an anticalin scaffold derived from human lipocalin (such as those described in WO 2015 / 104406); an avimer scaffold or protein fragment derived from the A domain of low density related protein (LRP) and / or very low density lipoprotein receptor (VLDLR); a finomer scaffold or portion thereof of the SH3 domain of FYN tyrosine kinase; a Kunitz domain scaffold or portion thereof of a Kunitz-type protease inhibitor, such as human trypsin inhibitor, aprotinin (bovine pancreatic trypsin inhibitor), Alzheimer's amyloid precursor protein, and tissue factor pathway inhibitor; It will be appreciated that the scaffolds may include knottin scaffolds (cysteine ​​knot miniproteins), such as those based on trypsin inhibitors derived from Staphylococcus elaterium; affibody scaffolds or all or part of the Z domain of Staphylococcus aureus (S. aureus) protein A; β-hairpin mimetic scaffolds; designed ankyrin repeat protein (DARPin) scaffolds or artificial protein scaffolds based on ankyrin repeat (AR) proteins; or any scaffold derived from or based on human transferrin, human CTLA-4, human crystallin, and human ubiquitin. For example, the binding site of human transferrin for the human transferrin receptor can be diversified to create a diverse library of transferrin mutants, some of which have acquired affinity for different antigens. See, e.g., Ali et al. (1999) J. Biol. Chem. 274:24066-24073.The part of human transferrin that is not involved in binding to receptor remains unchanged, and serves as a scaffold for providing mutant binding site, like the framework region of an antibody.Then, this library is screened against the target antigen of interest, similar to antibody library, and according to the method described herein, to identify the mutant with the best selectivity and affinity for the target antigen.See, for example, Hey et al. (2005) TRENDS Biotechnol.23(10):514-522.

[0226] constant region In some embodiments, a targeting group, such as an antibody or antigen-binding portion thereof, or other targeting group, comprises an antibody constant region. In some embodiments, the constant region is a fully human constant region. In some embodiments, the constant region is a humanized constant region. In some embodiments, the constant region is a non-human constant region. An immunoglobulin constant region refers to a heavy or light chain constant region. Human heavy and light chain constant region amino acid sequences are known in the art. The constant region may be of any suitable type selected from immunoglobulin classes, such as IgA, IgD, IgE, IgG, and IgM. Some immunoglobulin classes can be further divided into isotypes, e.g., IgG1, IgG2, IgG3, IgG4, or IgA1 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 (κ) or lambda (λ).

[0227] In some embodiments, the constant region may have an IgG isotype. 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 constant region 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 constant region is of the IgG1 isotype and has the amino acid sequence set forth in SEQ ID NO:2. In some embodiments, the constant region is of the κ isotype and has the amino acid sequence set forth in SEQ ID NO:3.

[0228] Furthermore, targeting groups comprising antibodies or antigen-binding portions thereof or non-antibody scaffolds may be part of a larger molecule formed by covalent or non-covalent binding of the antibody or antigen-binding portion to one or more other proteins or peptides. Examples of such targeting groups include the use of streptavidin core regions to prepare 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 hexahistidinyl tags ("hexahistidinyl tags" disclosed as SEQ ID NO:4) to produce bivalent biotinylated scFv molecules (Kipriyanov, SM, et al. (1994) Mol. Immunol. 31:10471058).

[0229] Fc domain modifications that alter effector function In some embodiments, the Fc region or Fc domain of a targeting group, such as an antibody or antigen-binding portion thereof, or a non-antibody scaffold, 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, "does not substantially bind" refers to weak or no binding to the selected Fcγ receptor. In some embodiments, "does not substantially bind" refers to at least a 1000-fold reduction in binding affinity to the Fcγ receptor (i.e., an increased Kd). In some embodiments, the Fc domain or region is Fc null. As used herein, "Fc null" refers to an Fc region or Fc domain that exhibits weak or no binding to any of the Fcγ receptors. In some embodiments, the Fc null domain or region exhibits at least a 1000-fold reduction in binding affinity to the Fcγ receptor (i.e., an increased Kd).

[0230] In some embodiments, the Fc domain has reduced or substantially no effector function activity. As used herein, "effector function activity" refers to antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent 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 affect ADCC, ADCP, or CDC). As used herein, "substantially no effector function" refers to at least a 1000-fold reduction in effector function activity compared to a wild-type or reference Fc domain.

[0231] In some embodiments, the Fc domain has reduced or no ADCC activity. As used herein, reduced or no ADCC activity refers to an Fc domain having at least 10, at least 20, at least 30, at least 50, at least 100, or at least 500 times less ADCC activity.

[0232] In some embodiments, the Fc domain has reduced or no CDC activity. As used herein, reduced or no CDC activity refers to an Fc domain having at least 10, at least 20, at least 30, at least 50, at least 100, or at least 500 times less CDC activity.

[0233] To confirm the reduction / elimination of ADCC and / or CDC activity, in vitro and / or in vivo cytotoxicity assays can be performed. For example, an Fc receptor (FcR) binding assay can be performed to confirm that the antibody does not bind to Fcγ receptors (and therefore likely lacks ADCC activity). NK cells, the primary cells mediating ADCC, express only FcγRIII, whereas 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 to assess ADCC activity of a molecule of interest are described in U.S. Pat. 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)); U.S. Pat. No. 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361 (1987)). Alternatively, non-radioactive assay methods may be employed (see, e.g., the ACTI™ non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. Mountain View, Calif.); and CytoTox 96™ non-radioactive cytotoxicity assay (Promega, Madison, Wis.)). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al., Proc. Nat'l Acad. Sci. USA 95:652-656 (1998).

[0234] C1q binding assay can also be carried out to confirm that antibody or Fc domain or region cannot bind to C1q and therefore lacks or has low CDC activity.For example, see the C1q and C3c binding ELISA in WO2006 / 029879 and WO2005 / 100402.CDC assay can be carried out to evaluate complement activation (for example, see 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)).

[0235] In some embodiments, the Fc domain has reduced or no ADCP activity. As used herein, reduced or no ADCP activity refers to an Fc domain having at least 10, at least 20, at least 30, at least 50, at least 100, or at least 500 times less ADCP activity.

[0236] ADCP binding assays can also be performed to confirm that an antibody or Fc domain or region lacks or has reduced ADCP activity. See, for example, US20190079077 and US20190048078.

[0237] Targeting groups, such as antibodies or antigen-binding portions thereof or non-antigenic scaffolds, with reduced effector function activity include substitutions of one or more of the Fc region residues, such as, for example, 238, 265, 269, 270, 297, 327, and 329, according to the EU numbering of Kabat (see, e.g., U.S. Pat. No. 6,737,056). Such Fc variants include those with substitutions at two or more of amino acid positions 265, 269, 270, 297, and 327 according to the EU numbering of Kabat, including the so-called "DANA" Fc variant in which residues 265 and 297 are substituted with alanine (see, e.g., U.S. Pat. No. 7,332,581). Certain antibody variants with reduced binding to FcRs are also known. (See, e.g., U.S. Pat. No. 6,737,056; WO2004 / 056312, and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001).) Targeting groups such as antibodies or antigen-binding portions thereof or non-antibody scaffolds that contain such amino acid modifications can be prepared that have reduced binding to FcRs.

[0238] In some embodiments, the targeting group, such as an antibody or antigen-binding portion thereof, or a non-antibody scaffold, comprises 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 (EU numbering of residues). In some embodiments, the substitutions are L234A and L235A (LALA) according to Kabat EU numbering. In some embodiments, the Fc domain comprises D265A and / or P329G in an Fc region derived from a human IgG1 Fc region according to Kabat EU numbering. In some embodiments, the substitutions are L234A, L235A, and P329G (LALA-PG) in an Fc region derived from a human IgG1 Fc region according to Kabat EU numbering. (See, e.g., WO2012 / 130831). In some embodiments, the substitutions are L234A, L235A, and D265A (LALA-DA), according to the EU numbering of Kabat, in an Fc region derived from a human IgG1 Fc region.

[0239] In some embodiments, modifications are made to the Fc region that result in altered (i.e., either decreased) C1q binding and / or complement-dependent cytotoxicity (CDC), as described in U.S. Pat. No. 6,194,551, WO 99 / 51642, and Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

[0240] Methods of Making Antibodies and Antigen-Binding Moieties and Other Targeting Groups In various embodiments, targeting groups such as antibodies and their antigen-binding portions can be produced in cell lines derived from humans, mice, or other animals. Recombinant DNA expression can be used to produce antibodies and their antigen-binding portions. This allows for the production of antibodies and various antigen-binding portions (including fusion proteins) in a selected host species. Production of antibodies and their antigen-binding portions in bacteria, yeast, transgenic animals, and chicken eggs also provides alternatives to cell-based manufacturing systems. A major advantage of transgenic animals is the potential for high yields from renewable resources.

[0241] Nucleic acid molecules encoding the amino acid sequence of a targeting group, such as an antibody or its antigen-binding portion, can be prepared by various methods known in the art. These methods include, but are not limited to, preparing synthetic nucleotide sequences encoding antibodies or antigen-binding portions. In addition, oligonucleotide-mediated (or site-directed) mutagenesis, PCR-mediated mutagenesis, and cassette mutagenesis can be used to prepare nucleotide sequences encoding antibodies or antigen-binding portions. At least the nucleic acid sequence encoding an antibody or its antigen-binding portion or polypeptide described herein can be recombined with vector DNA according to conventional techniques, such as restriction enzyme digestion to provide blunt or cohesive ends for ligation, suitable ends, filling in suitable cohesive ends, alkaline phosphatase treatment to avoid undesired ligation, and ligation with an appropriate ligase or other techniques known in the art. Techniques for such manipulations 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 antibodies or antigen-binding portions thereof, or VH or VL polypeptides thereof.

[0242] As used herein, the terms "nucleic acid" or "nucleic acid sequence" or "polynucleotide sequence" or "nucleotide" refer to a polymeric molecule incorporating units of ribonucleic acid, deoxyribonucleic acid, or analogs thereof. A nucleic acid may be either single-stranded or double-stranded. A single-stranded nucleic acid may be a single strand of denatured double-stranded DNA. In some embodiments, a nucleic acid may be cDNA, e.g., a nucleic acid lacking introns.

[0243] Nucleic acid molecules such as DNA contain nucleotide sequences containing transcriptional and translational regulatory information, and are said to be "capable of expressing" a polypeptide when such sequences are "operably linked" to a nucleotide sequence encoding the polypeptide. Operable linkage is a linkage in which the regulatory DNA sequence and the DNA sequence desired to be expressed (e.g., an antibody or its antigen-binding portion) are linked in a manner that allows gene expression of recoverable amounts of the polypeptide or antigen-binding portion. The exact nature of the regulatory regions required for gene expression can vary from organism to organism, as is well known in the art. See, for example, Sambrook et al., 1989; Ausubel et al., 1987-1993.

[0244] Thus, expression of targeting groups, such as antibodies or antigen-binding portions thereof, 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 can be human, primate, hamster, rabbit, rodent, bovine, porcine, ovine, equine, caprine, canine, or feline, although other mammalian cells may also be used. Furthermore, in vivo synthesis of ubiquitin-transmembrane polypeptide fusion proteins can be achieved, for example, by use of the yeast ubiquitin hydrolase system. Fusion proteins thus produced can be processed in vivo or purified and processed in vitro, allowing for the synthesis of antibodies or antigen-binding portions thereof described herein with specific amino-terminal sequences. Furthermore, problems associated with retention of the methionine residue from the start codon during direct yeast (or bacterial) expression can be avoided. (See, e.g., Sabin et al., 7 Bio / Technol. 705 (1989); Miller et al., 7 Bio / Technol. 698 (1989)). Recombinant antibodies, or antigen-binding portions thereof, can be obtained using any of a range of yeast gene expression systems that incorporate promoter and termination elements from actively expressed genes encoding glycolytic enzymes that are produced in large amounts when yeast is grown in glucose-rich medium. Known glycolytic genes can also provide highly efficient transcriptional control signals. For example, the promoter and terminator signals of the phosphoglycerate kinase gene can be used.

[0245] Production of antibodies or antigen-binding portions in insects can be achieved, for example, by infecting the insect host with a baculovirus that has been engineered to express the polypeptide by methods known to those of skill in the art. See Ausubel et al., 1987-1993.

[0246] In some embodiments, the introduced nucleic acid sequence (encoding the antibody or antigen-binding portion thereof, or the polypeptide) is incorporated into a plasmid or viral vector capable of autonomous replication in the recipient host cell. Any of a wide variety of vectors can be used for this purpose and are known and available to those of skill in the art. See, e.g., 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 that do not contain the vector; the copy number of the vector desired in the particular host; and whether it is desirable to be able to "shuttle" the vector between host cells of different species.

[0247] Exemplary prokaryotic vectors known in the art include plasmids capable of replicating in Escherichia coli (E. coli). Other gene expression elements useful for expressing DNA encoding an antibody or antigen-binding portion thereof 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 murine 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 (Okayama et al., 1983), and (c) polyadenylation sites, such as those present in SV40 (Okayama et al., 1983). As described in Liu et al. and Weidle et al. (51 Gene 21 (1987) below), DNA genes encoding immunoglobulins can be expressed using the SV40 early promoter and its enhancer, the mouse immunoglobulin heavy chain promoter enhancer, the SV40 late region mRNA splicing sequence, the rabbit S globin intervening sequence, the immunoglobulin and rabbit S globin polyadenylation sites, and the SV40 polyadenylation element as expression elements.

[0248] 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.

[0249] In some embodiments, for 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 of a mouse immunoglobulin heavy chain enhancer and a viral LTR enhancer, as well as polyadenylation and transcription termination regions. In another embodiment, a DNA sequence encoding another protein is combined with the above expression elements to achieve protein expression in mammalian cells.

[0250] Each coding region or gene fusion is incorporated or inserted into an expression vector. The nucleotides encoding the antibody, or its antibody polypeptide or antigen-binding portion, are then transfected alone or co-transfected with polynucleotides encoding the VH and VL chain coding regions into recipient cells capable of expressing the variable region or antigen-binding portion thereof. The transfected recipient cells are cultured under conditions that allow expression of the incorporated coding regions, and the expressed antibody chains or complete antibody or antigen-binding portion are recovered from the culture.

[0251] In some embodiments, nucleic acids containing coding regions encoding antibodies or antigen-binding portions thereof are incorporated into separate expression vectors and then co-transfected 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 a bacterial system and a second selection gene designed for selection in a eukaryotic system, with each vector carrying a set of coding regions. This strategy first directs the production of nucleotide sequences in a bacterial system, resulting in vectors that allow amplification. The DNA vectors thus generated and amplified in the bacterial host are then used to co-transfect eukaryotic cells, allowing for the selection of co-transfected cells carrying the desired transfected nucleic acids (e.g., containing the heavy and light chains of an antibody). Non-limiting examples of selection genes for use in bacterial systems include genes that confer ampicillin resistance and genes that confer chloramphenicol resistance. Selection genes for use in eukaryotic transfectants include the xanthine guanine phosphoribosyltransferase gene (designated gpt) and the phosphotransferase gene from Tn5 (designated neo). Alternatively, fused nucleotide sequences encoding the VH and VL chains can be incorporated onto the same expression vector.

[0252] For transfection of expression vectors and production of antibodies or antigen-binding portions thereof, the recipient cell line can be a Chinese hamster ovary cell line (e.g., DG44) or a myeloma cell. Myeloma cells can synthesize, assemble, and secrete immunoglobulins encoded by the transfected immunoglobulin genes and possess the immunoglobulin glycosylation machinery. 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 transfected genes. Myeloma cells can be grown in culture or in the peritoneal cavity of mice, where secreted immunoglobulins can be obtained from the ascites fluid.

[0253] Expression vectors encoding antibodies or antigen-binding portions thereof can be introduced into suitable host cells by any of a variety of suitable means, including biochemical means such as transformation, transfection, protoplast fusion, calcium phosphate precipitation, and in combination with polycations such as diethylaminoethyl (DEAE) dextran, and mechanical means such as electroporation, direct microinjection, and particle bombardment, as known to those skilled in the art (see, e.g., Johnston et al., 240 Science 1538 (1988)).

[0254] Yeast has several advantages over bacteria for 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 that can be used for the production of desired proteins in yeast. Yeast recognizes leader sequences in cloned mammalian gene products and secretes polypeptides bearing leader sequences (i.e., prepolypeptides). See, e.g., Hitzman et al., 11th Intl. Conf. Yeast, Genetics & Molec. Biol. (Montpelier, France, 1982).

[0255] Yeast gene expression systems can be routinely evaluated for the degree of production, secretion, and stability of antibodies, assembled antibodies, and their antigen-binding sites. A variety of yeast gene expression systems are available that incorporate promoter and termination elements of genes encoding glycolytic enzymes that are produced in large quantities when yeast is cultured in glucose-rich media. Known glycolytic genes can also provide highly efficient transcriptional control signals. For example, the promoter and terminator signals of the phosphoglycerate kinase (PGK) gene can be utilized. 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, e.g., U.S. Patent Application Publication No. 2006 / 0270045A1.

[0256] Bacterial strains can also be used as hosts for the production of the antibody molecules or antigen-binding portions thereof described herein. Examples of suitable host cells include E. coli K12 strains such as E. coli W3110, Bacillus species, Enterobacteriaceae such as Salmonella typhimurium or Serratia marcescens, and various Pseudomonas species. In connection with these bacterial host cells, plasmid vectors containing replicon and control sequences derived from species compatible with the host cell are used. The vector contains a replication site as well as specific genes that allow phenotypic selection in transformed cells. Various approaches are available for evaluating expression plasmids for the production of antibodies and their antigen-binding portions in bacteria (see Glover, 1985; Ausubel, 1987, 1993; Sambrook, 1989; Colligan, 1992-1996).

[0257] Host mammalian cells can be grown in vitro or in vivo and provide post-translational modifications to immunoglobulin molecules, including removal of leader peptides, folding and assembly of VH and VL chains, glycosylation of antibody molecules, and secretion of functional antibodies and / or antigen-binding sites.

[0258] Mammalian cells that can be useful as hosts for producing antibody proteins include cells of fibroblast origin, such as Vero cells or CHO-K1 cells, in addition to the cells of lymphoid origin described above. Exemplary eukaryotic cells that can be used to express immunoglobulin polypeptides include, but are not limited to, 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. In some embodiments, a particular eukaryotic host cell is selected based on its ability to make desired post-translational modifications to the heavy and / or light chains. For example, in some embodiments, CHO cells produce polypeptides with higher levels of sialylation than the same polypeptides produced in 293 cells.

[0259] In some embodiments, one or more antibodies or antigen-binding portions thereof can be produced in vivo in an animal that has been engineered or transfected with one or more nucleic acid molecules encoding the polypeptides, according to any suitable method.

[0260] In some embodiments, the antibody or antigen-binding portion thereof is 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).

[0261] Many vector systems are available for expressing VH and VL chains in mammalian cells (see Glover, 1985). Complete antibodies can be obtained according to various approaches. As discussed above, VH and VL chains, and optionally the associated constant regions, can be co-expressed in the same cell to achieve intracellular association and linkage of the VH and VL chains and produce a complete tetrameric H2L2 antibody or antigen-binding portion thereof. Co-expression can occur using the same or different plasmids in the same host. Cells expressing both chains can be directly selected by incorporating nucleic acids encoding the VH and VL chains or their antigen-binding portions into the same plasmid, which can then be introduced into cells. Alternatively, a plasmid encoding one chain, e.g., the VL chain, can be first introduced into cells, followed by the introduction of a second VH chain plasmid containing a selectable marker into the resulting cell line. Cell lines producing the antibody or antigen-binding portion thereof by either route can be transfected with plasmids encoding additional copies of the peptide, VH, VL, or VH and VL chains in combination with additional selectable markers to generate cell lines with improved properties, such as increased production of assembled antibody or antigen-binding portion thereof, or improved stability of the transfected cell line.

[0262] Furthermore, plants have emerged as a convenient, safe, and economical alternative expression system for recombinant antibody production based on large-scale cultivation of microorganisms or animal cells. Antibodies or antigen-binding portions thereof can be expressed in plant cell cultures or in conventionally cultivated plants. Expression in plants can be systemic, restricted to intracellular plastids, or restricted to seeds (endosperm). See, for example, U.S. Patent Application Publication No. 2003 / 0167531; U.S. Patent No. 6,080,560; U.S. Patent No. 6,512,162; and WO0129242. Several plant-derived antibodies have reached advanced stages of development, including clinical trials (see, for example, Biolex, NC).

[0263] In the case of intact antibodies, the variable regions of the antibody (VH and VL regions) are typically linked to at least a portion of a human immunoglobulin constant region (Fc) or domain. Human constant region DNA sequences can be isolated according to well-known procedures from various human cells, such as immortalized B cells (WO87 / 02671). The antibody can contain both light and heavy chain constant regions. The heavy chain constant region can include CH1, hinge, CH2, CH3, and optionally CH4 regions. In some embodiments, the CH2 domain can be deleted or omitted.

[0264] Techniques described for the production of single-chain antibodies (see, e.g., U.S. Pat. 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 by reference in their entireties) can be adapted to produce single-chain antibodies that specifically bind to a target antigen. Single-chain antibodies are formed by linking the heavy and light chain variable regions of the Fv region via an amino acid bridge, resulting in a single-chain polypeptide. Techniques for the assembly of functional Fv portions in E. coli can also be used (see, e.g., Skerra et al., Science 242:1038-1041 (1988); which is incorporated by reference in its entirety).

[0265] In some embodiments, the antigen-binding portion comprises one or more scFvs. An scFv can be a fusion protein of the variable regions of an antibody's heavy chain (VH) and light chain (VL) linked by a short linker peptide, for example, consisting of 10 to about 25 amino acids. The linker typically contains many glycines for flexibility and serine or threonines for solubility, and can 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. scFv antibodies are described, for example, in Houston, J.S., Methods in Enzymol. 203 (1991) 46-96. Methods for producing 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.

[0266] In some embodiments, the antigen-binding portion of a single domain antibody is an antibody portion consisting of a single monomeric variable antibody region. Single domain antibodies can be derived from the variable domain of an antibody heavy chain from a camelid (e.g., a nanobody or VHH region). Additionally, single domain antibodies can be autonomous human heavy chain variable domains (aVH) or VNAR portions from sharks (see, e.g., Hasler et al., Mol. Immunol. 75:28-37, 2016).

[0267] Techniques for producing single domain antibodies (DABs or VHHs) are known in the art, for example, as disclosed 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). VHHs can have strong antigen-binding capabilities and can interact with epitopes that are inaccessible to conventional VH-VL pairs (see, e.g., Muyldermans et al., 2001). Alpaca serum IgG contains approximately 50% heavy chain-only IgG antibodies (HCAbs) of camelid origin (see, e.g., Maass et al., 2007). Alpacas can be immunized with antigens, and VHHs that bind to and neutralize the target antigen 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 be used to isolate antibody fragments using standard biopanning techniques well known in the art (see, e.g., Maass et al., 2007).

[0268] Techniques for producing multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (e.g., Milstein and Cuello, Nature 305: 537 (1983)), WO 93 / 08829, and Traunecker et al., EMBO J. 10: 3655 (1991)), and "knob-in-hole" engineering (see, e.g., U.S. Pat. No. 5,731,168; Carter (2001), J Immunol Methods 248, 7-15). Multispecific antibodies can be made by manipulating electrostatic steering effects to create antibody Fc heterodimeric molecules (see, e.g., WO 2009 / 089004A1); cross-linking two or more antibodies or antigen-binding portions thereof (see, e.g., U.S. Pat. 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" technology to create bispecific antibody portions (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv (scFv) dimers (see, e.g., Gruber et al., J. Immunol., 152:5368 (1994)); and, for example, by preparing trispecific antibodies as described in Tutt et al. J. Immunol. 147: 60 (1991).

[0269] Engineered antibodies with three or more functional antigen-binding sites, including "octopus antibodies," can also be targeting groups (see, e.g., U.S. Patent Application Publication No. 2006 / 0025576A1).

[0270] In some embodiments, the targeting group comprises different antigen-binding sites fused to one or the other of the two subunits of the Fc domain; thus, the two subunits of the Fc domain may be comprised in two non-identical polypeptide chains. Recombinant co-expression of these polypeptides and subsequent dimerization results in several possible combinations of the two polypeptides. Therefore, to improve the yield and purity of bispecific molecules in recombinant production, it may be advantageous to introduce modifications to the Fc domain of the targeting group that promote the association of the desired polypeptides.

[0271] Generally, this method involves replacing one or more amino acid residues at the interface of the two Fc domains with charged amino acid residues, thereby making homodimer formation electrostatically unfavorable, but heterodimer formation electrostatically favorable.

[0272] In some embodiments, the targeting group is a "bispecific T cell engager" or BiTE (see, e.g., WO2004 / 106381, WO2005 / 061547, WO2007 / 042261, and WO2008 / 119567). This approach utilizes two antibody variable domains arranged on a single polypeptide. For example, the single polypeptide chain can include two single-chain Fv (scFv) portions, each having a variable heavy (VH) and variable light (VL) domain, separated by a polypeptide linker of sufficient length to allow intramolecular association between the two domains. The single polypeptide further includes a polypeptide spacer sequence between the two scFvs. Each scFv recognizes a different epitope, and these epitopes can be specific for different proteins, both of which are bound by the BiTE.

[0273] Because the bispecific T cell engager is a single polypeptide, it can be expressed using any prokaryotic or eukaryotic cell expression system known in the art, such as a CHO cell line. However, specific purification techniques (see, e.g., EP 1691833) may be required to separate the monomeric bispecific T cell engager from other multimeric species that may have biological activity other than the intended activity of the monomer. In one exemplary purification scheme, a solution containing the secreted polypeptide is first subjected to metal affinity chromatography, and the polypeptide is eluted with a gradient of imidazole concentration. The eluate is further purified using anion exchange chromatography, and the polypeptide is eluted with a gradient of sodium chloride concentration. Finally, the eluate is subjected to size exclusion chromatography to separate the monomer from the multimeric species. In some embodiments, the targeting group is a bispecific antibody consisting of a single polypeptide chain comprising two single-chain FV moieties (scFVs) fused to each other by a peptide linker.

[0274] In some embodiments, the targeting group is multispecific, such as IgG-scFv. IgG-scFv formats include 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 making 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.

[0275] Igg-like dual variable domain antibodies (DVD-Ig) are described in Wu et al., 2007, Nat Biotechnol 25:1290-97; Hasler et al., Mol. Immunol. 75:28-37, 2016, and WO 08 / 024188 and WO 07 / 024715. Triomabs are described in Chelius et al., MAbs 2(3):309-319, 2010. 2-in-1-IgGs are described in Kontermann et al., Drug Discovery Today 20(7):838-847, 2015. Tanden antibodies or TandAbs are described in Kontermann et al., ibid. ScFv-HSA-scFv antibodies are also described in Kontermann et al., ibid.

[0276] Intact (e.g., whole) antibodies, their dimers, individual light and heavy chains, or antigen-binding portions thereof can be recovered and purified by known techniques, such as immunoabsorption or immunoaffinity chromatography, chromatographic methods such as HPLC (high performance liquid chromatography), ammonium sulfate precipitation, gel electrophoresis, or any combination thereof. See generally, Scopes, Protein Purification (Springer-Verlag, NY, 1982). Substantially pure antibodies or antigen-binding portions thereof having at least about 90% to 95% homogeneity are advantageous, particularly for pharmaceutical uses, as are those having 98% to 99% or more homogeneity. Once purified, partially or to the desired homogeneity, intact antibodies or antigen-binding portions thereof can be used therapeutically or in the development and implementation of assay procedures, immunofluorescence staining, and the like. See generally, Vols. I & II Immunol. Meth. (Lefkovits & Pernis, eds., Acad. Press, NY, 1979 and 1981).

[0277] Complex In some embodiments, conjugates are provided that include a targeting group described herein attached to a drug-linker described herein. In some embodiments, conjugates are provided that include a targeting group described herein attached to a drug-linker described herein (e.g., Formula (A)). For example, in some embodiments, the targeting group is selected from an antibody or antigen-binding portion thereof. In more specific embodiments, the targeting group is a monoclonal antibody, Fab, Fab', F(ab'), Fv, disulfide-linked Fc, scFv, single-domain antibody, diabody, bispecific antibody, or multispecific antibody. In more specific embodiments, the targeting group is a diabody, DART, anticalin, affibody, avimer, DARPin, or adnectin. In some embodiments, the targeting group is monospecific. In some embodiments, the targeting group is bivalent. In some embodiments, the targeting group is bispecific.

[0278] In some embodiments, a conjugate is provided comprising a targeting group attached to a drug-linker as described above, wherein the average drug loading of the conjugate (p load ) is about 1 to about 8, about 2, about 4, about 6, about 8, about 10, about 12, about 14, about 16, about 3 to about 5, about 6 to about 8, or about 8 to about 16. In some cases, p load is approximately 1. In some cases, p load is approximately 2. In some cases, p load is approximately 4. In some cases, p load is about 8.

[0279] In some embodiments, the following: TIFF2026504840000216.tif48156TIFF2026504840000217.tif237157TIFF2026504840000218.tif205156TI FF2026504840000219.tif216150TIFF2026504840000220.tif199157TIFF2026504840000221.tif242159TIFF 2026504840000222.tif248165TIFF2026504840000223.tif191162TIFF2026504840000224.tif213166TIFF20 26504840000225.tif226165TIFF2026504840000226.tif197164TIFF2026504840000227.tif247168TIFF2026 504840000228.tif254163TIFF2026504840000229.tif255162TIFF2026504840000230.tif206164TIFF20265 04840000231.tif254165TIFF2026504840000232.tif226165TIFF2026504840000233.tif227166TIFF2026504 a complex selected from TIFF2026504840000234.tif216167TIFF2026504840000235.tif237162TIFF2026504840000236.tif220163TIFF2026504840000237.tif228163TIFF2026504840000238.tif234164TIFF2026504840000239.tif55159 is provided; where Ab is a targeting group and n is p load is.

[0280] In some embodiments, the conjugate is a stereoisomer of one of the above structures.

[0281] In some embodiments, the conjugate described above is provided, wherein the targeting group specifically binds to a target molecule. In more specific embodiments, the target molecule is CD19, CD20, CD30, CD33, CD70, LIV-1, or EGFRv3. In more specific embodiments, the target molecule is CD19, CD20, CD30, CD33, CD70, LIV-1 EGFRv3, or HER2. For example, in some embodiments, the targeting group is selected from scFv1-ScFv2, ScFv12-Fc-scFv22, IgG-scFv, DVD-Ig, triomab / quadroma, two-in-one IgG, scFv2-Fc, TandAb, and scFv-HSA-scFv.

[0282] In another aspect, there is provided a conjugate as described above, wherein the target molecule is a cancer-associated antigen. For example, in some embodiments, the target molecule is CD19, CD20, CD30, CD33, CD38, CA125, HER2, MUC-1, prostate-specific membrane antigen (PSMA), CD44 surface adhesion molecule, mesothelin (MLSN), carcinoembryonic antigen (CEA), epidermal growth factor receptor (EGFR), EGFRvIII, vascular endothelial growth factor receptor 2 (VEGFR2), high molecular weight melanoma-associated antigen (HMW-MAA), MAGE-A1, IL-13R-a2, GD2, 1p19q, ABL1, AKT1, ALK, APC, AR, ATM, BRAF, BRCA1, BRCA2, cKIT, cMET, CSF1R, CTNNB1, FGFR1, FGFR2, FLT3, GNA11, GNAQ, GNAS, HRAS, IDH1, IDH2, JAK2, KDR (VEGFR2), KRAS, MGMT, MGMT-Me, MLH1, MPL, NOTCH1, NRAS, PDGFRA, Pgp, PIK3CA, PR, PTEN, RET, RRM1, SMO, SPARC, TLE3, TOP2A, TOPO1, TP53, TS, TU BB3, VHL, CDH1, ERBB4, FBXW7, HNF1A, JAK3, NPM1, PTPN11, RB1, SMAD4, SMARCB1, STK1, MLH1, MSH2, MSH6, PMS2, ROS1, ERCC1, 5T4(TPBG), B7-H3, CCR 7, CD105, CD22, CD46, CD47, CD56, CD70, CD71, CD79b, CDH6, CLDN6, CLDN18.2, CLEC12A, DLL3, DR5, ERBB3 (HER3), EPCAM, FOLR1, IGF1R, IL2RA (CD25), IL3RA, ITGB6, LIV-1, LRRC15, mesothelin (MSLN), NaPi2b (SLC34A2), nectin-4, PTK7, ROR1, SEZ6, SLC44A4, SLITRK6, tissue factor (TF), TROP2, or B7-H4.

[0283] In another embodiment, the targeting group is selected from the group consisting of rituximab (Rituxan®), trastuzumab (Herceptin®), pertuzumab (Perjeta®), bevacizumab (Avastin®), ranibizumab (Lucentis®), cetuximab (Erbitux®), alemtuzumab (Campath®), panitumumab (Vectibix®), ibritumomab Conjugates as described above are provided which are antibodies, or fragments thereof, comprising tiusetan (Zevalin®), tositumomab (Vexar®), ipilimumab, zalutumumab, dalotuzumab, figitumumab, ramucirumab, galiximab, faretuzumab, ocrelizumab, ofatumumab (Arzerra®), tositumumab, ibritumomab, CD20 antibody 2F2 (HuMax-CD20), 7D8, IgM2C6, IgG1 2C6, 11B8, B1, 2H7, LT20, 1FS, or AT80, daclizumab (Zenapax®), or anti-LHRH receptor antibodies including clones A9E4, F1G4, AT2G7, GNRH03, or GNRHR2.

[0284] Drug Loading A conjugate may contain one or more drug units per targeting group. The number of drug units per targeting group is called the drug loading. The drug loading of a conjugate is defined as the average number of drug units (drug molecules (e.g., cytotoxic agents)) per targeting group (e.g., antibody or antigen-binding moiety or non-antibody scaffold or non-antibody protein) in the conjugate, p load For example, p load is about 4, the average drug loading considering all targeting groups (e.g., antibodies or antigen-binding moieties or non-antibody scaffolds or non-antibody proteins) present in the composition is about 4. In some embodiments, p load is in the range of about 3 to about 5, about 3.6 to about 4.4, or about 3.8 to about 4.2. load may be about 3, about 4, or about 5. In some embodiments, p loadIn some embodiments, p is in the range of about 6 to about 8, more preferably about 7.5 to about 8.4. load may be about 6, about 7, or about 8. In some embodiments, p load is in the range of about 8 to about 16.

[0285] The average number of drug units per targeting group (e.g., antibody or antigen-binding moiety or non-antibody scaffold) in a preparation can be characterized by conventional methods such as UV, mass spectrometry, capillary electrophoresis (CE), and HPLC. load The quantitative distribution of the complex in terms of p may also be determined. In some cases, the p load Separation, purification, and characterization of homogeneous conjugates with a particular value of β from conjugates with different drug loadings can be achieved.

[0286] Conjugation of Drug-Linkers to Antibodies, Antigen-Binding Sites, and Other Binding Agents (Including Non-Antibody Scaffolds) Techniques for linking a drug unit to a targeting group (such as an antibody or its antigen-binding portion or a non-antibody scaffold) via a linker 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 attached to the drug unit (e.g., a cytotoxic agent, an immunomodulator, or other agent), and then the drug-linker is attached to the targeting group (e.g., an antibody or its antigen-binding portion or a non-antibody protein scaffold). In some embodiments, the linker is first attached to the targeting group (e.g., an antibody or its antigen-binding portion or a non-antibody protein scaffold), and then the drug unit is attached to the linker. In the following description, the term drug-linker is used to illustrate the attachment of a linker or drug-linker to a targeting group; those skilled in the art will recognize that the attachment method selected can be determined depending on the linker and the drug unit. In some embodiments, the Drug Unit is attached to the targeting group via a linker in a manner that reduces the activity of the Drug Unit until the Drug Unit is released from the conjugate (e.g., by hydrolysis, by proteolysis, or by a cleaving agent).

[0287] Generally, the conjugates can be prepared by several routes using organic chemical reactions, conditions, and reagents known to those skilled in the art, including: (1) the reaction of a bivalent linker with a nucleophilic group of a targeting group (e.g., an antibody or its antigen-binding portion, or a non-antibody protein scaffold) to form a targeting group-linker intermediate via a covalent bond, followed by reaction with a drug unit; and (2) the reaction of a bivalent linker with a nucleophilic group of a drug unit to form a drug-linker via a covalent bond, followed by reaction with a nucleophilic group of a targeting group. An exemplary method for preparing conjugates via the latter route is described in U.S. Patent No. 7,498,298, which is incorporated herein by reference.

[0288] Nucleophilic groups on targeting groups such as antibodies, antigen-binding moieties, and other binding substances (including non-antibody scaffolds) include, but are not limited to, (i) N-terminal amine groups, (ii) side-chain amine groups, such as lysine, (iii) side-chain thiol groups, such as cysteine, and (iv) sugar hydroxyl groups or amino groups on glycosylated antibodies. Amine, thiol, and hydroxyl groups are nucleophilic and can react with electrophilic groups on linkers 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, carboxyls, and maleimide groups. Certain targeting groups, such as antibodies (and antigen-binding moieties and other binding substances (including non-antibody scaffolds)), have reducible interchain disulfides, i.e., cysteine ​​bridges. Antibodies (and antigen-binding moieties and other binding agents, including non-antibody scaffolds) can be made more reactive for conjugation to linkers by treatment with a reducing agent, such as DTT (dithiothreitol) or tricarbonylethylphosphine (TCEP), such that the antibody is fully or partially reduced. Thus, each cysteine ​​bridge theoretically forms two reactive thiol nucleophiles. Additional nucleophilic groups can be introduced into targeting groups, such as antibodies (and antigen-binding moieties and other binding agents, including non-antibody scaffolds), through modification of lysine residues, for example, by reacting the lysine residue with 2-iminothiolane (Traut's reagent) to convert the amine to a thiol. Reactive thiol groups can also be introduced into targeting groups (such as antibodies and antigen-binding moieties and other binding agents, including non-antibody scaffolds)) by introducing one, two, three, four, or more cysteine ​​residues (e.g., by preparing antibodies, antigen-binding moieties, and other binding agents, including non-antibody scaffolds, that contain one or more non-naturally occurring cysteine ​​amino acid residues).

[0289] Conjugates can also be generated by the reaction of an electrophilic group on the targeting group, such as an aldehyde or ketone carbonyl group, with a nucleophilic group on a linker reagent. Useful nucleophilic groups on linker reagents include, but are not limited to, hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxyl, and arylhydrazide. In some embodiments, an antibody (or antigen-binding portion thereof or other binding agent, including non-antibody scaffolds) is modified to introduce an electrophilic moiety capable of reacting with a nucleophilic substituent on a linker. In another embodiment, the sugars of a glycosylated antibody can be oxidized, e.g., with a periodate oxidation reagent, to form an aldehyde or ketone group that can react with an amine group on a linker. The resulting imine Schiff base group can form a stable bond or can be reduced, e.g., with a borohydride reagent, to form a stable amine bond. In one embodiment, reaction of the carbohydrate moiety of a glycosylated antibody with either galactose oxidase or sodium metaperiodate can provide carbonyl (aldehyde and ketone) groups in the antibody (or antigen-binding portion thereof, or other binding substance, including non-antibody scaffolds) that can react with appropriate groups on a linker (see, e.g., Hermanson, Bioconjugate Techniques). In another embodiment, targeting groups such as antibodies containing N-terminal serine or threonine residues can be reacted with sodium metaperiodate, resulting in the generation of an aldehyde in place of the first amino acid (Geoghegan & Stroh, (1992) Bioconjugate Chem. 3:138-146; US 5,362,852). Such aldehydes can react with a linker.

[0290] Exemplary nucleophilic groups on a Drug Unit, such as a cytotoxic agent, 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 electrophilic group on a Linker, including (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.

[0291] In some embodiments, the drug-linker is attached to an interchain cysteine ​​residue of an antibody (or antigen-binding portion thereof or other binding agent, including non-antibody scaffolds). See, e.g., WO2004 / 010957 and WO2005 / 081711. In such embodiments, the linker typically contains a maleimide group for attachment to a cysteine ​​residue of an interchain disulfide. In some embodiments, the linker or drug-linker is attached to a cysteine ​​residue of an antibody or antigen-binding portion thereof, as described in U.S. Pat. Nos. 7,585,491 or 8,080,250. The drug loading of the resulting conjugate is typically in the range of 1-8 or 1-16.

[0292] In some embodiments, the linker or drug-linker is attached to a lysine or cysteine ​​residue of an antibody (or antigen-binding portion thereof or other binding agent), as described in WO 2005 / 037992 or WO 2010 / 141566. The drug loading of the resulting conjugate is typically in the range of 1 to 8.

[0293] In some embodiments, engineered cysteine ​​residues, polyhistidine sequences, glycoengineered tags, or transglutaminase recognition sequences can be used for site-specific attachment of a linker or drug-linker to an antibody or antigen-binding portion thereof or other binding agent (including non-antibody scaffolds).

[0294] In some embodiments, the drug-linker is attached to an engineered cysteine ​​residue in an Fc residue other than the interchain disulfide. In some embodiments, the drug-linker is attached to 118, 221, 224, 227, 228, 230, 231, 223, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 330, 331, 332, 333, 334, 335, 336, 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, and / or linked to engineered cysteines 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, and / or linked to light chain positions 106, 108, 142 (light chain), 149 (light chain), and / or V205. An exemplary substitution for site-specific linkage using an engineered cysteine ​​is S239C (see, e.g., US20100158909; Fc region numbering is according to the EU index).

[0295] In some embodiments, the linker or drug-linker is attached to one or more introduced cysteine ​​residues of an antibody (or antigen-binding portion thereof or other binding agent, including non-antibody scaffolds), as described in WO2006 / 034488, WO2011 / 156328, and / or WO2016040856.

[0296] In some embodiments, exemplary substitutions for site-specific conjugation using bacterial transglutaminase are N297S or N297Q in the Fc region. In some embodiments, the linker or drug-linker is attached to a glycan or modified glycan of an antibody or antigen-binding moiety or a glycoengineered antibody (or other binding agent, including non-antibody scaffolds). See, e.g., WO2017 / 147542, WO2020 / 123425, WO2020 / 245229, WO2014 / 072482, WO2014 / / 065661, WO2015 / 057066, and WO2016 / 022027, the disclosures of which are incorporated herein by reference.

[0297] In some embodiments, the linker or drug-linker is attached to an antibody, antigen-binding moiety, or other binding agent (including non-antibody scaffolds) via a sortase A linker, which can be generated by the sortase A enzyme fusing an LPXTG recognition motif (SEQ ID NO: 5) to an N-terminal GGG motif to regenerate a native amide bond.

[0298] In some embodiments, the linker or drug-linker is attached to an antibody, antigen-binding moiety, or other binding agent (including non-antibody scaffolds) using SMARTag technology, in which a bioorthogonal aldehyde handle is introduced into the aldehyde-containing formylglycine (fGly) through oxidation of a cysteine ​​residue embedded in a specific peptide sequence (CxPxR). This enzymatic modification is carried out by formylglycine generating enzyme (FGE). See, e.g., Liu et al., Methods Mol. Biol. 2033:131-147 (2019).

[0299] In some embodiments, the linker or drug-linker is attached to an antibody, antigen-binding moiety, or other binding agent (including non-antibody scaffolds) using cysteine ​​conjugation with quaternized vinyl- and alkynyl-pyridine reagents. See, e.g., Matos et al., Angew Chem. Int. Ed. Engl. 58:6640-6644 (2019).

[0300] In another embodiment, the linker or drug-linker is attached to the antibody, antigen-binding moiety, or other binding agent (including non-antibody scaffolds) using bismaleimide, C-lock, or K-lock methods.

[0301] Pharmaceutical Compositions Another aspect of the conjugate relates to a composition comprising an active ingredient, including any of the conjugates described herein. In some embodiments, the composition is a pharmaceutical composition. As used herein, the term "pharmaceutical composition" refers to an active ingredient in combination with a pharmaceutically acceptable carrier accepted for use in the pharmaceutical industry. The term "pharmaceutically acceptable" is used herein to refer to compounds, substances, compositions, carriers, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit-risk ratio.

[0302] The preparation of pharmacological compositions containing active ingredients dissolved or dispersed therein is well understood in the art and need not be limited based on any particular formulation. Typically, such compositions are prepared as injectable solutions or suspensions; however, solid forms suitable for rehydration or suspension in liquid prior to use can also be prepared. Preparations can also be emulsified or provided as liposomal compositions. The complex can be mixed with an excipient that is pharmaceutically acceptable and compatible with the active ingredient and in an amount appropriate for use in the therapeutic methods described herein. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol, etc., and combinations thereof. In addition, if desired, pharmaceutical compositions can contain minor amounts of auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and the like, which enhance or maintain the effectiveness of the active ingredient (e.g., the complex).

[0303] The pharmaceutical compositions described herein can contain pharmaceutically acceptable salts of the components therein.Pharmaceutically acceptable salts include, for example, inorganic acids such as hydrochloric acid or phosphoric acid, or acid addition salts (formed with the free amino group of polypeptide) formed with organic acids such as acetic acid, tartaric acid, mandelic acid, etc. The salts formed with free carboxylic acid groups 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, histidine, prodecane, etc.

[0304] Physiologically acceptable carriers are well known in the art. An exemplary liquid carrier is a sterile aqueous solution such as phosphate-buffered saline, which contains an active ingredient (e.g., a complex) and water and may contain a buffer such as sodium phosphate, saline, or both at a physiological pH. 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 liquid phases in addition to or in addition to water. Examples of such additional liquid phases are glycerin, vegetable oils such as cottonseed oil, and water-oil emulsions. The amount of active agent effective in treating a particular disease or condition will vary depending on the nature of the disease or condition and can be determined by standard clinical techniques.

[0305] In some embodiments, the pharmaceutical composition comprising the conjugate may be a lyophilized agent.

[0306] In some embodiments, a syringe containing a therapeutically effective amount of the conjugate is provided.

[0307] Treatment method In some embodiments, a method of treating a subject is provided, comprising administering to the subject a conjugate described herein or a pharmaceutical composition described herein. For example, in some embodiments, the subject has cancer or an autoimmune disease, and the conjugate binds to a target antigen associated with the cancer or autoimmune disease.

[0308] In some embodiments, a method for treating cancer is provided, comprising administering the conjugate. In some embodiments, a subject is in need of treatment for cancer and / or malignant tumors. In some embodiments, the method is for treating a subject with cancer or malignant tumors.

[0309] The methods described herein include administering a therapeutically effective amount of a conjugate to a subject with cancer or a malignant tumor. As used herein, the terms "therapeutically effective amount," "effective amount," or "effective dose" refer to an amount of a conjugate that provides a therapeutic benefit in the treatment, management, or prevention of recurrence of cancer or a malignant tumor, e.g., an amount that provides a statistically significant reduction in at least one symptom, sign, or marker of the tumor or malignant tumor. Determining a therapeutically effective amount is well within the capabilities of one of ordinary skill in the art. Generally, a therapeutically effective amount may vary depending on the subject's medical history, age, condition, and sex, as well as the severity and type of the condition in the subject, and the administration of other pharmaceutically active agents.

[0310] The terms "cancer" and "malignancy" refer to the uncontrolled growth of cells that interferes with the normal function of bodily organs and systems. Cancer or malignant tumors can be primary or metastatic, i.e., the cancer invades and spreads to tissues distant from the original tumor site. A "tumor" refers to the uncontrolled growth of cells that interferes with the normal function of bodily organs and systems. A subject with cancer refers to a subject with objectively measurable cancer cells present in the subject's body. This definition includes benign and malignant tumors, as well as potentially dormant tumors and micrometastases. When cancer migrates from its original location and spreads to other vital organs, it can lead to the death of the subject through impaired function of the affected organ. Hematological malignancies (hematopoietic cancers), such as leukemia and lymphoma, can, for example, outcompete the normal hematopoietic compartment in a subject, resulting in hematopoietic failure (in the form of anemia, thrombocytopenia, and neutropenia) and ultimately death.

[0311] Examples of cancer include, but are not limited to, carcinoma, lymphoma, germinoma, sarcoma, and leukemia. More specific examples of such cancers include basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and CNS cancer, breast cancer (e.g., triple-negative breast cancer), peritoneal cancer, cervical cancer; bile duct cancer, choriocarcinoma, chondrosarcoma, colon and rectal cancer (colorectal cancer), connective tissue cancer, digestive system 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. , intraepithelial neoplasia, renal 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's lymphoma and non-Hodgkin's lymphoma, melanoma, mesothelioma, myeloma, neuroblastoma, oral cancer (e.g., lip cancer, tongue cancer, oral cavity cancer, and pharyngeal cancer), ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma , respiratory system cancer, salivary gland cancer, sarcoma, skin cancer, squamous cell carcinoma, testicular cancer, thyroid cancer, uterine or endometrial cancer, uterine intussusception cancer, urinary system cancer, vulvar cancer; other carcinomas and sarcomas, B-cell lymphomas (low-grade / follicular non-Hodgkin's 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 These include, but are not limited to, NHL, bulky disease NHL, mantle cell lymphoma, AIDS-related lymphoma, and Waldenstrom's macroglobulinemia), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myeloblastic leukemia, and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal blood vessel growth associated with phakomatosis, edema (such as that associated with brain tumors), and Meigs' syndrome.

[0312] The methods herein are contemplated to reduce tumor size or tumor burden in a subject and / or reduce metastasis in a subject. In various embodiments, tumor size in a subject is reduced by about 25-50%, about 40-70%, or about 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%.

[0313] As used herein, "subject" refers to a human or an animal. Typically, an animal is a vertebrate such as a primate, a rodent, a livestock or a game animal. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaque monkeys, e.g., rhesus monkeys. Rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Livestock and game animals include cattle, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cats, canine species, e.g., dogs, foxes, wolves, avian 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.

[0314] Preferably, the subject is a mammal. The mammal may be, but is not limited to, a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse, or a cow. Non-human mammals can be advantageously used as subjects, for example, to represent animal models of various cancers. 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 a human.

[0315] In some embodiments, the subject has previously been diagnosed with or identified as having cancer and is in need of treatment, but need not have already been treated for cancer. In some embodiments, the subject may also be a subject who has not previously been diagnosed with cancer in need of treatment. In some embodiments, the subject may be a subject who exhibits one or more risk factors for the condition or one or more complications associated with cancer, or a subject who does not exhibit risk factors. In particular, a subject "in need" of 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" a condition refers to a subject who has been diagnosed as being at risk of developing the condition or at risk of developing the condition again.

[0316] As used herein, the terms "treat," "treatment," "treating," or "amelioration" when used in reference to a disease, disorder, or condition refer to therapeutic treatment for a condition, where the objective is to reverse, alleviate, improve, inhibit, slow, or halt the progression or severity of the symptom or condition. The term "treating" 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 the improvement of symptoms or markers, but also the 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 results include, but are not limited to, a reduction in cancer cells in a subject, alleviation of one or more symptoms, a reduction in the extent of the defect, a stabilization (i.e., not worsening) of the cancer or malignant tumor condition, a delay or slowing of tumor growth and / or metastasis, and an increase in lifespan compared to that expected in the absence of treatment. As used herein, the term "administering" refers to providing a conjugate described herein to a subject by a method or route that results in binding of the conjugate to cancer or malignant cells. Similarly, a pharmaceutical composition comprising a conjugate described herein can be administered by any appropriate route that results in effective treatment in the subject.

[0317] The dosage range of the conjugate varies depending on potency and includes an amount sufficient to produce the desired effect, e.g., tumor growth delay or tumor size reduction. The dosage should not be so large as to cause unacceptable side effects. Generally, the dosage will vary depending on the age, condition, and sex of the subject and can be determined by one of ordinary skill in the art. The dosage can also be adjusted by an individual physician if any complications arise. In some embodiments, the dosage range is between 0.1 mg / kg body weight and 10 mg / kg body weight. In some embodiments, the dosage range is between 0.5 mg / kg body weight and 15 mg / kg body weight. In some embodiments, the dosage range is between 0.5 mg / kg body weight and 5 mg / kg body weight. Alternatively, the dosage range can be titrated to maintain a blood concentration between 1 μg / mL and 1000 μg / mL. For systemic administration, a subject can receive a therapeutic amount, for example, 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.

[0318] The administration of the above doses can be repeated. In a preferred embodiment, the above doses are administered weekly, every other week, every three weeks, or monthly for several weeks or months. The duration of treatment varies depending on the subject's clinical course and response to treatment.

[0319] In some embodiments, the dose may be about 0.1 mg / kg to about 100 mg / kg. In some embodiments, the dose may be about 0.1 mg / kg to about 25 mg / kg. In some embodiments, the dose may be about 0.1 mg / kg to about 20 mg / kg. In some embodiments, the dose may be about 0.1 mg / kg to about 15 mg / kg. In some embodiments, the dose may be about 0.1 mg / kg to about 12 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 100 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 25 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 20 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 15 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 12 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 10 mg / kg.

[0320] In some embodiments, the dose may be administered intravenously. In some embodiments, the intravenous administration may be an infusion administered over a period of about 10 minutes to about 4 hours. In some embodiments, the intravenous administration may be an infusion administered over a period of about 30 minutes to about 90 minutes.

[0321] 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 about every two weeks. In some embodiments, the dose may be administered about every three weeks. In some embodiments, the dose may be administered every four weeks.

[0322] In some embodiments, the subject is administered about 2 to about 10 total doses. In some embodiments, the subject is administered 4 total doses. In some embodiments, the subject is administered 5 total doses. In some embodiments, the subject is administered 6 total doses. In some embodiments, the subject is administered 7 total doses. In some embodiments, the subject is administered 8 total doses. In some embodiments, the subject is administered 9 total doses. In some embodiments, the subject is administered 10 total doses. In some embodiments, the subject is administered more than 10 total doses.

[0323] Pharmaceutical compositions containing the conjugates can be administered in unit doses. The term "unit dose" when used in reference to pharmaceutical compositions refers to physically discrete units suitable as single administrations to subjects, each unit containing a predetermined amount of active substance (e.g., conjugate) calculated to produce a desired therapeutic effect in association with the required physiologically acceptable diluent, i.e., carrier or vehicle.

[0324] In some aspects, the conjugates described herein can be used in methods comprising administering the conjugate to a subject in need thereof, such as a subject with an autoimmune disease.

[0325] In some embodiments, methods for treating autoimmune disease are provided, comprising administering a conjugate described herein. In some embodiments, the subject is in need of treatment for the autoimmune disease. The methods described herein comprise administering a therapeutically effective amount of a conjugate to a subject with an autoimmune disease. As used herein, the terms "therapeutically effective amount," "effective amount," or "effective dose" refer to an amount of a conjugate described herein that provides a therapeutic benefit in the treatment, management, or prevention of recurrence of an autoimmune disease, e.g., an amount that provides a statistically significant reduction in at least one symptom, sign, or marker of an autoimmune disease. Determining a therapeutically effective amount is well within the capabilities of one of ordinary skill in the art. Generally, a therapeutically effective amount may vary depending on the subject's medical history, age, condition, and sex, the severity and type of the condition in the subject, and the administration of other pharmaceutically active agents.

[0326] The term "autoimmune disease" refers to an immune disorder characterized by inappropriate activation of immune cells (e.g., lymphocytes or dendritic cells) and that interferes with the normal function of the body's organs and systems. 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, Sjogren's syndrome, type 1 diabetes, primary biliary cirrhosis, Wegener's granulomatosis, fibromyalgia, polymyositis, dermatomyositis, polyendocrine deficiency, Schmidt's syndrome, autoimmune uveitis, and amyloidosis. Dison's disease, adrenalitis, thyroiditis, Hashimoto's disease, autoimmune thyroid disease, pernicious anemia, gastric atrophy, chronic hepatitis, lupus hepatitis, atherosclerosis, subacute cutaneous lupus erythematosus, hypoparathyroidism, Draisler's syndrome, autoimmune thrombocytopenia, idiopathic thrombocytopenic purpura, hemolytic anemia, pemphigus vulgaris, pemphigus, dermatitis herpetiformis, alopecia areata, pemphigoid, scleroderma, progressive systemic sclerosis, CREST syndrome (calcinosis, Raynaud's disease, esophageal dysmotility, sclerodactyly, and telangiectasia), male and female autoimmune infertility, ankylosing spine Spondylitis, ulcerative colitis, mixed connective tissue disease, polyarteritis nodosa, systemic necrotizing vasculitis, atopic dermatitis, atopic rhinitis, Goodpasture's syndrome, Chagas' disease, sarcoidosis, rheumatic fever, asthma, habitual abortion, antiphospholipid syndrome, farmer's lung, erythema multiforme, post-open heart surgery syndrome, Cushing's syndrome, autoimmune chronic active hepatitis, bird fancier's disease, toxic epidermal necrolysis, Alport's syndrome, alveolitis, allergic alveolitis, fibrosing alveolitis, interstitial lung disease, erythema nodosum, pyoderma gangrenosum, transfusion reaction, Takayasu's arteritis, rheumatic polyarteritis Myalgia, temporal arteritis, schistosomiasis, giant cell arteritis, ascariasis, aspergillosis, Samter's syndrome, eczema, lymphomatoid granulomatosis, Behçet's disease, Kaplan's syndrome, Kawasaki disease, dengue fever, encephalomyelitis, endocarditis, endomyocardial fibrosis, endophthalmitis, erythema elevata, psoriasis, erythroblastosis fetalis, eosinophilic fasciitis, Shulman's syndrome, Felty's syndrome, filariasis, cyclitis, chronic cyclitis, metachronous cyclitis, Fuchs' cyclitis, IgA nephropathy, Henoch-Schönlein purpura, graft-versus-host disease, transplant rejection, cardiomyopathy,Including, but not limited to, Eaton-Lambert syndrome, relapsing polychondritis, cryoglobulinemia, Waldenstrom macroglobulinemia, Evans syndrome, and autoimmune hypogonadism.

[0327] In some embodiments, the methods described herein encompass the treatment of a disease of B lymphocytes (e.g., systemic lupus erythematosus, Goodpasture's syndrome, rheumatoid arthritis, and type 1 diabetes), a disease of Th1 lymphocytes (e.g., rheumatoid arthritis, multiple sclerosis, psoriasis, Sjogren's syndrome, Hashimoto's disease, Graves' disease, primary biliary cholangitis, Wegener's granulomatosis, tuberculosis, or graft-versus-host disease), or a disease of Th2 lymphocytes (e.g., atopic dermatitis, systemic lupus erythematosus, atopic asthma, rhinoconjunctivitis, allergic rhinitis, Omenn's syndrome, systemic sclerosis, or chronic graft-versus-host disease). Generally, diseases involving dendritic cells involve diseases of Th1 lymphocytes or Th2 lymphocytes.

[0328] As used herein, "subject" refers to a human or an animal. Typically, an animal is a vertebrate such as a primate, a rodent, a livestock or a game animal. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaque monkeys, e.g., rhesus monkeys. Rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Livestock and game animals include cattle, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cats, canine species, e.g., dogs, foxes, wolves, avian 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.

[0329] Preferably, the subject is a mammal. The mammal may be, but is not limited to, a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse, or a cow. Non-human mammals can be advantageously used as subjects that represent animal models of various autoimmune diseases, for example. 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 a human.

[0330] In some embodiments, a subject has previously been diagnosed with or identified as suffering from an autoimmune disease and is in need of treatment, but need not have already been treated for the autoimmune disease. In some embodiments, a subject may also be a subject who has not previously been diagnosed with an autoimmune disease in need of treatment. In some embodiments, a subject may exhibit one or more risk factors for the condition or one or more complications associated with an autoimmune disease, or may not exhibit risk factors. In particular, a subject "in need" of 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" a 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).

[0331] As used herein, the terms "treat," "treatment," "treating," or "amelioration" when used in reference to a disease, disorder, or condition refer to therapeutic treatment for a condition, where the objective is to reverse, alleviate, improve, inhibit, slow, or halt the progression or severity of the symptom or condition. The term "treating" 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 the improvement of symptoms or markers, but also the 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 results include, but are not limited to, a reduction in autoimmune cells in a subject, alleviation of one or more symptoms, a reduction in the degree of deficiency, stabilization of the autoimmune disease state (i.e., not worsening), a delay or slowing of the progression of the autoimmune disease, and an increase in lifespan compared to that expected in the absence of treatment. As used herein, the term "administering" refers to providing a conjugate described herein to a subject by a method or route that results in binding of the conjugate to target autoimmune cells. Similarly, a pharmaceutical composition comprising a conjugate described herein can be administered by any suitable route that results in effective treatment in a subject.

[0332] The dosage range of the conjugate varies depending on potency and includes an amount sufficient to produce the desired effect, e.g., slowing the progression of or reducing the symptoms of an autoimmune disease. The dosage should not be so large as to cause unacceptable side effects. Generally, the dosage will vary depending on the age, condition, and sex of the subject and can be determined by one of ordinary skill in the art. The dosage can also be adjusted by an individual physician if any complications arise. In some embodiments, the dosage range is between 0.1 mg / kg body weight and 10 mg / kg body weight. In some embodiments, the dosage range is between 0.5 mg / kg body weight and 15 mg / kg body weight. In some embodiments, the dosage range is between 0.5 mg / kg body weight and 5 mg / kg body weight. Alternatively, the dosage range can be titrated to maintain a blood concentration between 1 μg / mL and 1000 μg / mL. For systemic administration, a subject can receive a therapeutic amount, for example, 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.

[0333] The administration of the above doses can be repeated. In a preferred embodiment, the above doses are administered weekly, every other week, every three weeks, or monthly for several weeks or months. The duration of treatment varies depending on the subject's clinical course and response to treatment.

[0334] In some embodiments, the dose may be about 0.1 mg / kg to about 100 mg / kg. In some embodiments, the dose may be about 0.1 mg / kg to about 25 mg / kg. In some embodiments, the dose may be about 0.1 mg / kg to about 20 mg / kg. In some embodiments, the dose may be about 0.1 mg / kg to about 15 mg / kg. In some embodiments, the dose may be about 0.1 mg / kg to about 12 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 100 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 25 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 20 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 15 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 12 mg / kg. In some embodiments, the dose may be about 1 mg / kg to about 10 mg / kg.

[0335] In some embodiments, the dose may be administered intravenously. In some embodiments, the intravenous administration may be an infusion administered over a period of about 10 minutes to about 4 hours. In some embodiments, the intravenous administration may be an infusion administered over a period of about 30 minutes to about 90 minutes.

[0336] 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 about every two weeks. In some embodiments, the dose may be administered about every three weeks. In some embodiments, the dose may be administered every four weeks.

[0337] In some embodiments, the subject is administered about 2 to about 10 total doses. In some embodiments, the subject is administered 4 total doses. In some embodiments, the subject is administered 5 total doses. In some embodiments, the subject is administered 6 total doses. In some embodiments, the subject is administered 7 total doses. In some embodiments, the subject is administered 8 total doses. In some embodiments, the subject is administered 9 total doses. In some embodiments, the subject is administered 10 total doses. In some embodiments, the subject is administered more than 10 total doses.

[0338] The pharmaceutical composition containing the conjugate can be administered in a unit dose. The term "unit dose" when used in reference to a pharmaceutical composition refers to a physically discrete unit suitable as a single administration to a subject, each unit containing a predetermined quantity of active substance (e.g., conjugate) calculated to produce a desired therapeutic effect in association with the required physiologically acceptable diluent, i.e., carrier or vehicle.

[0339] In some embodiments, the conjugate, or any of these pharmaceutical compositions, is administered in conjunction with immunosuppressive therapy. In some embodiments, a method for improving therapeutic outcomes in a subject receiving immunosuppressive therapy is provided. The method generally includes administering an effective amount of immunosuppressive therapy to a subject with an autoimmune disease; and administering a therapeutically effective amount of a conjugate or pharmaceutical composition thereof that specifically binds to target autoimmune cells to the subject; wherein the therapeutic outcomes of the subject are improved compared to administering immunotherapy alone. In some embodiments, the conjugate is as described herein. In some embodiments, the improved therapeutic outcomes include a reduction in disease progression, alleviation of one or more symptoms, etc.

[0340] The present invention is further illustrated by the following embodiments, which should not be construed as limiting. Aspect 1 (a) Having 1 to 4 binding sites per drug unit and having the following structure (i) or (ii): linker unit with one of TIFF2026504840000240.tif39128; (b) at least one polar group comprising a polymer unit, optionally a sugar unit, optionally a carboxyl unit, or a combination thereof; and (c) optionally, a stretcher group having a binding site for a targeting group; or a stereoisomer or salt thereof, During the ceremony, α- is the attachment site to the enzyme-cleavable group; β- is the site of attachment to at least one polar group; δ- is H, a site of attachment to at least one of the Drug units, or a site of attachment to a linking group attached to at least one of the Drug units; the polymer units comprise polyamides, polyethers, or combinations thereof, wherein the polyethers comprise hydroxyl groups, polyhydroxyl groups, sugar groups, carboxyl groups, or combinations thereof; Each R a are independently H or C1-C6 alkyl; Each R b is independently halo, C 1~6 alkyl, a bond to at least one of the drug units, or a bond to at least one polar group; x is 0, 1, 2, 3, or 4; y is 0, 1, 2, or 3; R c is a bond, -C(O)-, -S(O)-, -SO2-, 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)-, -SO2-C1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, triazolyl-containing group, or a combination thereof; A linker compound or a stereoisomer or salt thereof. Aspect 2 The linker unit has the following structure (ia), (ii-a), or (iii-a): 2. The linker compound of embodiment 1, having one of the following: TIFF2026504840000241.tif96128, or a stereoisomer or salt thereof. Aspect 3 The linker unit has the following structure (ib), (ic), (id), (ie), or (if): 3. The linker compound of embodiment 1 or 2, having one of the following: TIFF2026504840000242.tif136132, or a stereoisomer or salt thereof. Aspect 4 The linker unit has the following structure (ii-b) or (iii-b): 2. The linker compound of embodiment 1, having TIFF2026504840000243.tif54128 or a stereoisomer or salt thereof. Aspect 5 The polar group has the formula: L3-N(CH2-(CH(XR)) k -X1(X2))2(X) or 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 1 to 10; L3 is the point of attachment to the rest of the polar group, The linker compound according to any one of embodiments 1 to 4. Aspect 6 At least one sugar unit has the following structure (XII) or (XIII): TIFF2026504840000244.tif73128 or a stereoisomer or salt thereof, During the ceremony, each R is independently selected from hydrogen, monosaccharides, disaccharides, and polysaccharides; m is 1 to 8; n is 0 to 4; The linker compound of embodiment 5. Aspect 7 .below: (a) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 (XX) or a stereoisomer, salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or C1-C3 alkylene; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle; n20 is 2 to 26; or (b) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NR 24 R 25 (XXI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or C1-C3 alkylene; R 24 and R 25 one of which is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;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-CH2-CH2-] n20 R 29 ] n21 -R 27 -NR 24 R 25 ] n27 (XXII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 26 and R 27 are arbitrary, and bonds, C1 to 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 12Alkylene-C(O)-, -NH-C1-C 12 Alkylene -C(O)- and -C(O)-C1 to C 12 alkylene-NH-; R 24 and R 25 one of which is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;R 24 and R 25 the other is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 and polyethylene glycol, optionally having 1 to 24 ethylene glycol subunits; or -NR 24 R 25 together form a C3-C8 heterocycle; Each R 29is optionally independently selected from —C(O)—, —NH—, —C(O)—C1-C6 alkylene-, —NH—C1-C6 alkylene-, —C1-C6 alkylene-NH—, —C1-C6 alkylene-C(O)—, —NH(CO)—C1-C6 alkylene-, —N(CH3)—(CO)—C1-C6 alkylene-, —NH(CO)NH—, and triazole; n20 is 2-26; n21 is 1 to 4; n27 is 1 to 4; or (d) ~R 20 -R 21 -[-C(R α )HC(O)-N(R N )-] n20 -R 22 -NR 24 R 25 (XXIII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 is a bond, 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 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)-; Each R α are independently H or -R 22 -NR 24 R 25 and; Each R N are independently H, C1-C6 alkyl, or -R 22 -NR24 R 25 and; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle; each n20 independently is 2 to 26; or (e) ~R 20 -R 21 -[-C(R α )HC(O)-N(R N )-] n20 -R 22 -CO2R 26 (XXIV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond, C1-C3 alkylene, or -C1-C3 alkylene [O-CH2-CH2-] n20 and; Each R α are independently H or -R 22 -NR 24 R 25 and; Each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and; R 24and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle; R 26 is H or C1-C4 alkyl; Each n20 is independently 2 to 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 )HC(O)-N(R N )-] n20 -R 22 -N-(R 23 -NR 24 R 25 )2(XXV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond, C1-C3 alkylene, or -C1-C3 alkylene-[O-CH2-CH2-] n20 and; Each R α are independently H or -R 22 -NR 24 R25 and; Each R N are independently H or C1-C6 alkyl; Each R 23 are independently C1-C6 alkylene; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; optionally substituted heteroaryl; optionally substituted carbocycle; substituted -C1-C8 alkyl; substituted -C(O)-C1-C8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle; Each n20 is independently 2 to 26; 7. The linker compound of any one of embodiments 1 to 6, comprising a polar group having a formula selected from: Aspect 8 .R 24 and R 25 and n is 0 or 1. The linker compound of embodiment 7, wherein both of Aspect 9 .R 24 and R 25 are each independently selected from H and a polyhydroxyl group, with the proviso that R 24 and R 25 and n is 0 or 1. The linker compound of embodiment 7 or 8, wherein both of Aspect 10 The linker compound of any one of embodiments 7 to 9, wherein the polyhydroxyl group is a linear monosaccharide optionally selected from a C6 or C5 sugar, a sugar acid, and an amino sugar. Aspect 11the C6 or C5 sugar is selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, gulose, 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; 11. The linker compound according to embodiment 10. Aspect 12 .below: TIFF2026504840000245.tif228120TIFF2026504840000246.tif253148TIFF2026504840000247.tif252134TIFF2026504840000248.tif234158TIFF2026504840000249.tif241159TIFF2026504840000250.tif229158TIFF2026504840000251.tif254158TIFF2026504840000252.tif153143, or a stereoisomer or salt thereof; wherein each R is independently H or alkyl; and each R 39 are independently selected from H, linear monosaccharides, and polyethylene glycols optionally having 1 to 24 ethylene glycol subunits; each n is independently 1 to 12; the wavy line represents R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 7. Aspect 13 .R 24 and R 25 9. The linker compound according to embodiment 7 or 8, wherein one of said monosaccharides is a linear monosaccharide and the other is a cyclic monosaccharide. Aspect 14 .below: TIFF2026504840000253.tif106128 or a stereoisomer or salt thereof; In the formula, R 41 is a cyclic monosaccharide; the wavy line is R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 7. Aspect 15 .R 24 and R 25 is independently a polyhydroxyl selected from cyclic monosaccharides, disaccharides, and polysaccharides. Aspect 16 .below: TIFF2026504840000254.tif156128 or a stereoisomer or salt thereof; In the formula, each R 45 is selected from H and a monosaccharide, a disaccharide, or a polysaccharide; R 46 is selected from cyclic monosaccharides, disaccharides, or polysaccharides; the wavy line represents R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 7. Aspect 17 .R 24 and R 25 are independently selected from a linear monosaccharide and a substituted linear monosaccharide, wherein the substituted linear monosaccharide is substituted with a monosaccharide, a disaccharide, or a polysaccharide. Aspect 18 .below: TIFF2026504840000255.tif152128 or a stereoisomer or salt thereof; In the formula, R 47 is a linear monosaccharide; each R 49 is selected from monosaccharides, disaccharides, and polysaccharides; the wavy line is R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 7. Aspect 19.R 24 and R 25 are independently selected from linear monosaccharides and substituted monosaccharides, wherein the substituted linear monosaccharides are substituted with one or more substituents selected from alkyl, O-alkyl, aryl, O-aryl, carboxyl, ester, or amide, and optionally further substituted with a monosaccharide, disaccharide, or polysaccharide. Aspect 20 .below: TIFF2026504840000256.tif81128 or a stereoisomer or salt thereof; In the formula, each R 42 are independently selected from linear monosaccharides and substituted linear monosaccharides; each R 43 are independently selected from alkyl, O-alkyl, aryl, O-aryl, carboxyl, ester, and amide; the wavy line represents R b The linker compound of any one of embodiments 1 to 7, wherein the linker compound is a bond to a hydroxyl group or to an enzyme-cleavable group. Aspect 21 .R 24 and R 25 is a -C(O)-polyhydroxyl group or a substituted -C(O)-polyhydroxyl group, and R 24 and R 25 the other is H, -C(O)-polyhydroxyl group, substituted -C(O)-polyhydroxyl group, polyhydroxyl group, or substituted polyhydroxyl group; wherein the substituted -C(O)-polyhydroxyl group and polyhydroxyl group are substituted with monosaccharide, disaccharide, polysaccharide, alkyl, -O-alkyl, aryl, carboxyl, ester, or amide, The linker compound according to any one of embodiments 7 to 8. Aspect 22 .below: TIFF2026504840000257.tif69128 or a stereoisomer or salt thereof; In the formula, the wavy line represents R b or to an enzyme-cleavable group, 22. The linker compound according to any one of embodiments 1 to 21. Aspect 23 .R 24 and R 25 are independently selected from H, substituted -C1-C8 alkyl, substituted -C1-C4 alkyl, or substituted -C1-C3 alkyl; with the proviso that R 24 and R 25 and wherein the substituted -C1-C8 alkyl, -C1-C4 alkyl, and -C1-C3 alkyl are substituted with hydroxyl and / or carboxyl. Aspect 24 .below: TIFF2026504840000258.tif235101 or a stereoisomer or salt thereof; In the formula, R 48 is selected from H, OH, CHOH, COOH, or -C1-C6 alkyl substituted with hydroxyl or carboxyl; the wavy line represents R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 7. Aspect 25 .R 24 and R 25 is selected from H, substituted —C(O)—C1-C8 alkyl, substituted —C(O)—C1-C4 alkyl, and substituted —C(O)—C1-C3 alkyl; and R 24 and R 25 is selected from substituted -C(O)-C1-C8 alkyl, substituted -C(O)-C1-C4 alkyl, substituted -C(O)-C1-C3 alkyl, substituted -C1-C8 alkyl, substituted -C1-C4 alkyl, and substituted -C1-C3 alkyl, wherein substituted -C(O)-C1-C8 alkyl, substituted -C(O)-C1-C4 alkyl, substituted -C(O)-C1-C3 alkyl, substituted -C1-C8 alkyl, -C1-C4 alkyl, and -C1-C3 alkyl is substituted with hydroxyl and / or carboxyl. Aspect 26 .below: TIFF2026504840000259.tif226121TIFF2026504840000260.tif21128 or a stereoisomer or salt thereof; In the formula, the wavy line represents R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 7. Aspect 27 .R 24 and R 25 is selected from H and optionally substituted aryl; provided that R 24 and R 25 The linker compound of any one of embodiments 6 to 8, wherein both of are not H. Aspect 28 .below: TIFF2026504840000261.tif69128 or a stereoisomer or salt thereof; In the formula, the wavy line represents R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 7. Aspect 29 .R 24 and R 25 The linker compound of embodiment 7 or 8, wherein together form an optionally substituted C3-C8 heterocycle or heteroaryl. Aspect 30 .The following structure: TIFF2026504840000262.tif12128 or a stereoisomer or salt thereof, wherein the wavy line represents R b The linker compound of any one of embodiments 1 to 7, wherein the linker compound is a bond to a hydroxyl group or to an enzyme-cleavable group. Aspect 31 .R 24 and R 25 is independently selected from H and a chelating agent, wherein the chelating agent is selected from —NR by alkylene, arylene, carbocyclo, heteroarylene, or heterocarbocylo; 24 R 25may be attached to the nitrogen of R 24 and R 25 and n is 0 or 1. The linker compound of embodiment 7 or 8, wherein both of Aspect 32 32. The linker compound of embodiment 31, wherein the chelator is selected from ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), triethylenetetraminehexaacetic acid (TTHA), benzyl-DTPA, 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), benzyl-DOTA, 1,4,7-triazacyclononane-N,N',N''-triacetic acid (NOTA), benzyl-NOTA, 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA), and N,N'-dialkyl-substituted piperazines. Aspect 33 .below: TIFF2026504840000263.tif75128 or a stereoisomer or salt thereof; In the formula, the wavy line represents R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 7. Aspect 34 Each monosaccharide independently a C5 or C6 sugar selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, gulose, idose talose, aldose, ketose, glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine; a sugar acid selected from gluconic acid, aldonic acid, uronic acid, and urosonic acid; or an amino sugar selected from glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine; 22. The linker compound according to any one of embodiments 6 to 21, selected from: Aspect 3535. The linker compound of any one of embodiments 1-34, wherein the attachment site is formed from a functional group of a 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, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, and protected forms thereof. Aspect 36 .below: (a) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -R 30 (XXX) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or a C1-C3 alkylene group; R 30 is an optionally substituted C3 to C 10 carbocycle; thiourea; optionally substituted thiourea; urea; optionally substituted urea; sulfamide; alkyl sulfamide; acylsulfamide; optionally substituted alkyl sulfamide; optionally substituted acylsulfamide; sulfonamide; optionally substituted sulfonamide; guanidine, including alkyl guanidine and aryl guanidine; phosphoramide; or optionally substituted phosphoramide; or R 30 represents azido, alkynyl, substituted alkynyl, -NH-C(O)-alkynyl, -NH-C(O)-alkynyl-R 65 cyclooctyne; selected from -NH-cyclooctyne, -NH-C(O)-cyclooctyne, or -NH-(cyclooctyne)2; where R 65is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2–26; (b) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NH-C(O)-R 31 (XXXI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 having; R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2–26; (c) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -C(O)NH-R 31 (XXXII) or a stereoisomer or salt thereof, wherein R 20 is part Rb or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch independently having 1 to 26 ethylene glycol subunits and each branch having R at its end 35 having; R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, and optionally substituted heteroaryl; n20 is 2–26; (d) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -C(O)NR 31 -R 22 -NR 24 R 25 (XXXIII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 31 is H or R 22 -NR 24 R 25 and; R 21 and R 22 each independently represents a bond or a C1-C3 alkylene group; R 24 and R 25are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 24 and R 25 Both of them cannot be H; n20 is 2–26; (e) ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -N(R 33 -R 31 )2(XXXIV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 having; R 33 is C1-C3 alkylene, C1-C3 alkylene-C(O), -C(O)-C1-C3 alkylene, or -C(O)-C1-C3 alkylene-C(O); R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2–26; (f) ~R 20 -(R 21 -[CH2-CH(OR34 )-CH2-O] n20 -R 36 ) n25 (XXXV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; Each R 21 are independently a bond, —O—, or a C1-C3 alkylene group; Each R 34 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 R 25 and; R N is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; or a substituted polyhydroxyl group, with the proviso that R 24 and R 25 Both of them cannot be H; Each R 36 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 are independently H or C1-C6 alkyl; R 44 and R45 are each 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 Both of them cannot be H; each n20 is independently 1 to 26; n25 is 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 -R 25 (XXXVI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 , R 22 and R 23 each independently represents a bond or a C1-C3 alkylene group; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 24 and R 25 Both of them cannot be H; each n20 independently represents 0 to 26, and each n21 independently represents 0 to 26, provided that at least one of n20 or n21 is 2 to 26; n22 is 1–5; each n23 is independently 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or a C1-C3 alkylene group; R N is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; or a substituted polyhydroxyl group, with the proviso that R 24 and R 25 Both of them cannot be H; Each R 34 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 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 are independently H or C1-C6 alkyl; R44 and R 45 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group; provided that R 44 and R 45 Both of them cannot be H; n20 is 2-26; n21 is 1 to 26; n25 is 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or a C1-C3 alkylene group; R N is H or C1-C4 alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; or a substituted polyhydroxyl group, with the proviso that R 24 and R 25 Both of them cannot be H; n20 is 2-26; 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond, 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 are independently H or -R 22 -NR 24 R 25 and; Each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; Optionally substituted heteroaryl; Optionally substituted carbocycle; Substituted -C1-C8 alkyl; Substituted -C(O)-C1-C8 alkyl; Chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle), provided that R 24 and R 25 Both of them cannot be H; each n20 independently is 0 to 26, provided that at least one n20 is 2 to 26; n25 is 1 or 2; or (k) TIFF2026504840000264.tif23128 or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 , R 22 and R 23 each independently represents a bond, 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 are independently H or -R 22 -NR 24 R 25 and; Each R N are independently H, C1-C6 alkyl, or -R 22 -NR 24 R 25 and; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; Optionally substituted heteroaryl; Optionally substituted carbocycle; Substituted -C1-C8 alkyl; Substituted -C(O)-C1-C8 alkyl; Chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 or -NR 24 R 25 together form a C3-C8 heterocycle), provided that R 24and R 25 Both of them cannot be H; R 26 is H or C1-C6 alkyl; each n20 independently is 0 to 26, provided that at least one n20 is 2 to 26; each n21 independently is 0 to 26, provided that at least one n21 is 2 to 26; 36. The linker compound of any one of embodiments 1 to 35, comprising a polar group having a formula selected from: Aspect 37 .below: ~R 20 -R 21 -[O-CH2-CH2] n20 -R 22 -NH-C(O)-R 31 (XXXI), ~R 20 -R 21 -[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) or a stereoisomer or salt thereof, During the ceremony, R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 each independently represents a bond or a C1-C3 alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 having; R 33is C1-C3 alkylene, -C1-C3 alkylene-C(O), -C(O)-C1-C3 alkylene, or -C(O)-C1-C3 alkylene-C(O); R 35 is azide, alkynyl, alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; the wavy (~) line is R 20 n20 is 2 to 26; The linker compound according to any one of embodiments 1 to 36. Aspect 38 .below: TIFF2026504840000265.tif221132, In the formula, R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; the wavy line represents R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 37. Aspect 39 .R b 39. The linker compound of any one of embodiments 36-38, wherein the attachment site to or to the enzyme-cleavable group is formed from a functional group of a precursor compound of the polar group, wherein the functional group is selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, and protected forms thereof. Aspect 40 .formula: ~R 20 -(R 43 -R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XL) or a stereoisomer or salt thereof, During the ceremony, R 20 is part R b or to an enzyme-cleavable group; R 41 and R 42 each independently represents a bond or C1-C6 alkylene; Each R 43 are independently a bond or C1 to 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-C 12 Alkylene, heteroarylene, heteroaryl-C1-C 12 Alkylene, heteroaryl-C1-C 12 Alkylene -C(O)- or -C(O)NR 46 R 47 where R 46 and R 47 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 is alkylene; R 44 and R 45are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; n40 is 2 to 26; n41 is 1-6; n42 is 1 to 6, The linker compound according to any one of embodiments 1 to 39. Aspect 41 .formula: ~R 20 -(R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLI) or a stereoisomer or salt thereof, During the ceremony, R 20 is part R b or to an enzyme-cleavable group; R 41 and R 42 each independently represents a bond or C1-C6 alkylene; R 43 is a bond or C1 to 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-C 12 Alkylene, heteroarylene, heteroaryl-C1-C 12 Alkylene, heteroaryl-C1-C 12Alkylene -C(O)- or -C(O)NR 46 R 47 where R 46 and R 47 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 is alkylene; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; n40 is 1 to 26; n41 is 1-6; n42 is 1 to 6, 41. The linker compound according to any one of embodiments 1 to 40. Aspect 42 .formula: ~R 20 -(R 41 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLII) or a stereoisomer or salt thereof, During the ceremony, R 20 is part R b or to an enzyme-cleavable group; R 41 and R 42 each independently represents a bond or C1-C3 alkylene; R 43 is a bond, C1-C6 alkylene, -NH-C1-C 12Alkylene, -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 where R 46 and R 47 One of the groups is H or C1-C6 alkylene, and the other is C1-C 12 is alkylene; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; n40 is 1 to 16; n41 is 1 to 4; n42 is 1 to 4, 42. The linker compound according to any one of embodiments 1 to 41. Aspect 43 .R 20 is formed from a functional group of a precursor compound of the polar group, wherein the functional group is selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, or a protected form thereof. Aspect 44 .R 20 But the following structure: TIFF2026504840000266.tif130159TIFF2026504840000267.tif212157 or a stereoisomer thereof, wherein R is H, a C1-C6 alkyl, or a polyhydroxyl group, and n is 0 to 12; TIFF2026504840000268.tif4128 is R b indicates the site of attachment to or to an enzyme-cleavable group, TIFF2026504840000269.tif4128 shows the polar group binding site to the rest of the molecule. The linker compound of any one of embodiments 7, 36, 37, and 40-42. Aspect 45 .R 20 But the following structure: TIFF2026504840000270.tif237161TIFF2026504840000271.tif107152 or a stereoisomer thereof, In the formula, n is 0 to 12; TIFF2026504840000272.tif4128 is R b indicates the site of attachment to or to an enzyme-cleavable group, TIFF2026504840000273.tif4128 shows the polar group binding site to the rest of the molecule. The compound of any one of embodiments 7, 36, 37, and 40-42. Aspect 46 .R 43 -(NR 44 R 45 ) n41 But the following structure: TIFF2026504840000274.tif129147 or a stereoisomer thereof, wherein R is H, C1-C6 alkyl, a polyhydroxyl group, or a substituted polyhydroxyl group; TIFF2026504840000275.tif4128 is the R to the rest of the polar group 43 indicates the binding site of 46. ​​The linker compound according to any one of embodiments 40 to 45. Aspect 47 .R 43 -(NR 44 R 45 ) n41 But the following structure: TIFF2026504840000276.tif157156 or a stereoisomer thereof, During the ceremony, TIFF2026504840000277.tif4128 is the R to the rest of the polar group 43 indicates the binding site of 46. ​​The linker compound according to any one of embodiments 40 to 45. Aspect 48 .-NR 44 R 45 But the following structure: TIFF2026504840000278.tif228161TIFF2026504840000279.tif72128 or a stereoisomer thereof, During the ceremony, TIFF2026504840000280.tif4128 is the polar group -NR to the rest of the 44 R 45 indicates the binding site of 48. The linker compound according to any one of embodiments 40 to 47. Aspect 49 Prior to attachment to the linker unit, the following structure: TIFF2026504840000281.tif113128TIFF2026504840000282.tif224154TIFF2026504840000283.tif229114TIFF2 026504840000284.tif217136TIFF2026504840000285.tif198157TIFF2026504840000286.tif208158TIFF2026504 840000287.tif198158TIFF2026504840000288.tif213158TIFF2026504840000289.tif195158TIFF202650484000 0290.tif212154TIFF2026504840000291.tif184155TIFF2026504840000292.tif197158TIFF2026504840000293.t if222158TIFF2026504840000294.tif244155TIFF2026504840000295.tif193157TIFF2026504840000296.tif168 156TIFF2026504840000297.tif201143TIFF2026504840000298.tif206155TIFF2026504840000299.tif239158TIF F2026504840000300.tif245103TIFF2026504840000301.tif21879TIFF2026504840000302.tif186160TIFF2026504840000303.tif241159TIFF2026504840000304.tif231160TIFF2026504840000305.tif33158, During the ceremony, 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 1 to 12; X is O, NR, or -CH2-; V is a bond or C1-C6 alkyl; R24 and R 25 one of which is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; Optionally substituted heteroaryl; Optionally substituted carbocycle; Substituted -C1-C8 alkyl; Substituted -C(O)-C1-C8 alkyl; Chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from;R 24 and R 25 the other is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; an optionally substituted C3-C 10 Carbocyclic ring; optionally substituted C1-C3 alkylene C3-C 10 Carbocycle; Optionally substituted heteroaryl; Optionally substituted carbocycle; Substituted -C1-C8 alkyl; Substituted -C(O)-C1-C8 alkyl; Chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 and polyethylene glycol, optionally having 1 to 24 ethylene glycol subunits; or -NR 24 R 25 together form a C3-C8 heterocycle, The linker compound according to any one of embodiments 1 to 48. Aspect 50 . below: (a) ~R 40 -(R 43 -R 41 -[O-CH2-CH2] n40 -R 46 -[O-CH2-CH2] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 )n42 (XLIII) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 41 and R 42 each independently represents a bond or C1-C6 alkylene; Each R 43 are independently bonded, C1 to 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-C 12 Alkylene, C1-C 12 Alkylene-NH-C(O)-, -heteroarylene, heteroaryl-C1-C 12 Alkylene, heteroaryl-C1-C 12 Alkylene -C(O)- or -C(O)NR 46 R 47 where R 46 and R 47 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 is alkylene; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; Each R 46 is independently -NR 50 -, -NR50 -C1~C6 alkylene-NR 50 -, -NR 50 -C(O)-NR 50 -S(O)2-NR 50 -, or -NR 50 -C(O)-C 1~6 alkylene-selected from; Each R 50 are independently selected from H, C1-C6 alkyl, or polyhydroxyl groups; n40 is 2-26; n41 is 1-6; n42 is 1-6; (b) ~R 40 -(R 51 -[O-CH2-CH2] n43 -R 52 -X1-R 55 -X2-R 53 -[O-CH2-CH2] n43 -R 54 -[X3-R 56 ] n44 -R 57 ) n45 (XLIV) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 51 , R 52 , R 53 and R 54 each independently represents a bond or C1-C6 alkylene; X1, X2 and X3 are each independently -NR N -C(O)- or -C(O)-NR N - and; Each R N independently represent H, C1-C6 alkyl, or polyhydroxyl groups; R 55 and R 56 each independently represents a divalent polyhydroxyl group; R 57is H, OH or C1-C6 alkyl; each n43 independently is 0 to 26, provided that at least one n43 is 1 to 26; n44 is 0-10; n45 is 1 or 2; or (c) ~R 40 -R 51 -[O-CH2-CH2] n43 -R 52 -N-(R 53 -X1-R 54 -[O-CH2-CH2] n43 -(NR 44 R 45 ))2 (XLV) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 51 , R 53 and R 54 each independently represents an optionally bonded or substituted C1-C6 alkylene; R 52 is a bond, C1-C6 alkylene, -C(O)-, or -OC(O)-; Each X1 is independently -NR N -C(O)- or -C(O)-NR N - and; Each R N independently represent H, C1-C6 alkyl, or polyhydroxyl groups; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; Each n43 is independently 2–26; 50. The linker compound of any one of embodiments 1 to 49, comprising a polar group having a formula selected from: Aspect 51 The following structure prior to attachment to the enzyme-cleavable group and / or to the linker unit: TIFF2026504840000306.tif38128TIFF2026504840000307.tif234161, During the ceremony, each R is independently H, an alkyl, or a polyhydroxyl group; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; each n is independently 1 to 12; The linker compound according to any one of embodiments 1 to 50. Aspect 52 . TIFF2026504840000308.tif212157 or a stereoisomer or salt thereof, During the ceremony, Each Y is independently R 76 or TIFF2026504840000309.tif18128; Each R 76 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH); Each R a and R b are independently H or R a and R b together with the carbon to which they are attached to form an oxo group; each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; each v is independently 1 to 6; Each * is R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 51. Aspect 53 . TIFF2026504840000310.tif216164 or a stereoisomer or salt thereof, During the ceremony, Each R 76 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v S(=O)2(OH); each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; each v is independently 1 to 6; Each * is R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 51. Aspect 54 . TIFF2026504840000311.tif218162 or a stereoisomer or salt thereof, During the ceremony, each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; Each * is R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 51. Aspect 55 .Y is R 76 53. The linker compound of embodiment 52, wherein: Aspect 56 .Y is 53. The linker compound of embodiment 52, wherein the linker compound is TIFF2026504840000312.tif18128. Aspect 57 .Each R a and R b 53. The linker compound of embodiment 52, wherein Aspect 58 .Ra and R b together with the carbon to which they are attached form an oxo group. Aspect 59 53. The linker compound of any one of embodiments 50 to 52, wherein q is 10 to 20. Aspect 60 55. The linker compound of any one of embodiments 52 to 54, wherein q is 12. Aspect 61 .below: TIFF2026504840000313.tif188158TIFF2026504840000314.tif229157TIFF2026504840000315.tif192157TIFF2026504840000316.t if222158TIFF2026504840000317.tif205157TIFF2026504840000318.tif255153TIFF2026504840000319.tif211157TIFF2026504840 000320.tif255156TIFF2026504840000321.tif212157TIFF2026504840000322.tif235158TIFF2026504840000323.tif222141TIFF20 26504840000324.tif201157TIFF2026504840000325.tif204157TIFF2026504840000326.tif207157TIFF2026504840000327.tif18914 8TIFF2026504840000328.tif225146TIFF2026504840000329.tif238146TIFF2026504840000330.tif238144TIFF2026504840000331. tif255148TIFF2026504840000332.tif219145TIFF2026504840000333.tif224126TIFF2026504840000334.tif186128TIFF202650484 0000335.tif21979TIFF2026504840000336.tif236157TIFF2026504840000337.tif190158TIFF2026504840000338.tif235159TIFF2026504840000339.tif248158TIFF2026504840000340.tif248158TIFF2026504840000341.tif224157, or a stereoisomer or salt thereof; During the ceremony, each Z is joined at *, Independently selected from TIFF2026504840000342.tif124159, each TIFF2026504840000343.tif2128 is R b or to an enzyme-cleavable group, The linker compound according to any one of embodiments 1 to 60. Aspect 62 .The following formula: TIFF2026504840000344.tif36128 or a stereoisomer or salt thereof, During the ceremony, (a) L 70 is selected from C1-C8 alkylene, C1-C8 alkylene-C(O)-, -C(O)-C1-C8 alkylene-, and -C(O)-C1-C8 alkylene-C(O)-, * is R b , to an enzyme-cleavable group, or to the remainder of a polar group; R 70 Ha~NR 71 (R 72 -R 73 ), where R 71 is H, C1~C 12 Alkyl, substituted C1-C 12 alkyl, or polyethylene glycol (which may have 1 to 12 ethylene glycol subunits); R 72 is a bond or is selected from an optionally substituted C1-C3 alkylene, an optionally substituted ether, an optionally substituted thioether, an optionally substituted ketone, an optionally substituted amide, a polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), an optionally substituted carbocycle, an optionally substituted aryl, or an optionally substituted heteroaryl; R 73is a carboxyl or polycarboxyl, wherein the polycarboxyl contains 1 to 10, or 1 to 6, or 1 to 4 carboxyl groups, and the carboxyl groups are interconnected by alkyl, alkylene, substituted alkyl, substituted alkylene, heteroalkyl, heteroalkylene, amino, and / or amido; or (b) L 70 is selected from C1-C8 alkylene, C1-C8 alkylene-C(O)-, -C(O)-C1-C8 alkylene-, and -C(O)-C1-C8 alkylene-C(O)-, * is R b , to an enzyme-cleavable group, or to the remainder of a polar group; R 70 Ha~NR 71 (R 75 - (R 73 )2), where R 71 is H, C1~C 12 Alkyl, substituted C1-C 12 alkyl, or polyethylene glycol (which may have 1 to 12 ethylene glycol subunits); R 75 is a branched optionally substituted C1-C3 alkylene, an optionally substituted ether, an optionally substituted thioether, an optionally substituted ketone, an optionally substituted amide, polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), an optionally substituted carbocycle, an optionally substituted aryl, or an optionally substituted heteroaryl, and each R 73 are independently carboxyl or polycarboxyl, wherein the polycarboxyl contains 1 to 10, or 1 to 6, or 1 to 4 carboxyl groups, and the carboxyl groups are interconnected by alkyl, alkylene, substituted alkyl, substituted alkylene, heteroalkyl, heteroalkylene, amino, and / or amido; or (c) L 70is selected from C1-C8 alkylene, C1-C8 alkylene-C(O)-, -C(O)-C1-C8 alkylene-, and -C(O)-C1-C8 alkylene-C(O)-, * is R b , to an enzyme-cleavable group, or to the remainder of a polar group; R 70 HA~N(R 74 -R 73 )(R 72 - R 73 ), where R 72 and R 74 are each independently selected from an optionally substituted C1-C3 alkylene, an optionally substituted ether, an optionally substituted thioether, an optionally substituted ketone, an optionally substituted amide, a polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), an optionally substituted carbocycle, an optionally substituted aryl, or an optionally substituted heteroaryl; and each R 73 are independently carbo...

Claims

1. (a) having 1 to 4 binding sites per Drug unit and having the following structure (i) or (ii): a linker unit having one of: (b) at least one polar group comprising a polymer unit, optionally a sugar unit, optionally a carboxyl unit, or a combination thereof; and (c) optionally, a stretcher group having a binding site for a targeting group; or a stereoisomer or salt thereof, comprising: During the ceremony, α- is the attachment site for the enzyme-cleavable group; β- is the attachment site to the at least one polar group; δ- is H, a site of attachment to at least one of the Drug units, or a site of attachment to a linking group attached to at least one of the Drug units; the polymer units comprise 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 are independently H or C 1 ~C 6 is alkyl; Each R b is independently halo, C 1～6 alkyl, a bond to at least one of the Drug units, or a bond 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 Alkylene, C 1 ~C 6 Alkenylene, C 1 ~C 6 alkynylene, triazolyl-containing groups, or combinations thereof; The linker compound or a stereoisomer or salt thereof.

2. The linker unit has the following structure (ia), (ii-a), or (iii-a):

2. The linker compound of claim 1, having one of:

3. The linker unit has the following structure (ib), (ic), (id), (ie), or (if):

2. The linker compound of claim 1, having one of:

4. The linker unit has the following structure (ii-b) or (iii-b):

2. The linker compound of claim 1, having:

5. The at least one polar group has the formula: L3-N(CH 2 -(CH(XR)) k -X 1 (X 2 )) 2 (X) or 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 is independently CH 2 and C(O); each X 2 is independently selected from H, OH, and OR; k is 1 to 10; L3 is the point of attachment to the remainder of the polar group; The linker compound of any one of claims 1 to 4.

6. At least one of the sugar units has the following structure (XII) or (XIII): or a stereoisomer or salt thereof, During the ceremony, each R is independently selected from hydrogen, monosaccharides, disaccharides, and polysaccharides; m is 1 to 8; n is 0 to 4; The linker compound of any one of claims 1 to 4.

7. below: (a) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -NR 24 R 25 (XX) or a stereoisomer, salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 alkylene; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted C 1 ~C 8 Alkyl; Substituted -C(O)-C 1 ~C 8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 Both of these cannot 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 alkylene; R 24 and R 25 one of which is H; polyhydroxyl group; substituted polyhydroxyl group; -C(O)-polyhydroxyl group; substituted -C(O)-polyhydroxyl group; substituted C 1 ~C 8 Alkyl; Substituted -C(O)-C 1 ~C 8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from: 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 26 and R 27 are arbitrary, and the bonds, C 1 ~C 12 Alkylene, -NH-C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkylene-NH-, -C 1 ~C 12 Alkylene-N(CH 3 )-, -C(O)-C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkylene-C(O)-, -NH-C 1 ~C 12 Alkylene -C(O)-, and -C(O)-C 1 ~C 12 independently selected from alkylene-NH-; R 24 and R 25 one of which is H; polyhydroxyl group; substituted polyhydroxyl group; -C(O)-polyhydroxyl group; substituted -C(O)-polyhydroxyl group; substituted C 1 ~C 8 Alkyl; Substituted -C(O)-C 1 ~C 8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from: R 24 and R 25 the other is selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; a substituted —C(O)-polyhydroxyl group; and a polyethylene glycol optionally having 1 to 24 ethylene glycol subunits; and R 28 is a sugar unit of formula (XII) or (XIII), provided that R 24 and R 25 Both of these cannot be H; Each R 29 is optional, -C(O)-, -NH-, -C(O)-C 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 independently selected from alkylene-, -NH(CO)NH-, and triazole; n20 is 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 is a bond 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 are independently H or -R 22 -NR 24 R 25 and Each R N are independently H, C 1 ~C 6 Alkyl, or -R 22 -NR 24 R 25 and R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted C 1 ~C 8 Alkyl; Substituted -C(O)-C 1 ~C 8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 Both of these cannot be H; each n20 independently is 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently bonded, C 1 ~C 3 Alkylene, or -C 1 ~C 3 Alkylene [O-CH 2 -CH 2 -] n20 and Each R α are independently H or -R 22 -NR 24 R 25 and Each R N are independently H, C 1 ~C 6 Alkyl, or -R 22 -NR 24 R 25 and R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted C 1 ~C 8 Alkyl; Substituted -C(O)-C 1 ~C 8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 Both of these cannot be H; R 26 is H or C 1 ~C 4 is alkyl; Each n20 is independently 2 to 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently bonded, C 1 ~C 3 Alkylene, or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n20 and Each R α are independently H or -R 22 -NR 24 R 25 and Each R N are independently H or C 1 ~C 6 is alkyl; Each R 23 is independently C 1 ~C 6 alkylene; R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted C 1 ~C 8 Alkyl; Substituted -C(O)-C 1 ~C 8 alkyl; chelating agent; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 Both of these cannot be H; each n20 is independently 2 to 26; 7. The linker compound of any one of claims 1 to 6, comprising a polar group having a formula selected from:

8. R 24 and R 25 are each independently selected from H and a polyhydroxyl group, and R 24 and R 25 8. The linker compound of claim 7, wherein both of are not H.

9. 9. The linker compound of claim 7 or 8, wherein the polyhydroxyl group is a linear monosaccharide optionally selected from a C6 or C5 sugar, a sugar acid, and an amino sugar.

10. the C6 or C5 sugar is selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, gulose, 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; 10. The linker compound of claim 9.

11. below: or a stereoisomer or salt thereof, wherein each R is independently H or alkyl; and each R 39 are independently selected from H, linear monosaccharides, and polyethylene glycols which may have 1 to 24 ethylene glycol subunits; each n is independently 1 to 12; the wavy lines represent R b or to said enzyme-cleavable group, The linker compound of any one of claims 7 to 10.

12. R 24 and R 25 9. The linker compound according to claim 7 or 8, wherein one of said monosaccharides is a linear monosaccharide and the other is a cyclic monosaccharide.

13. below: or a stereoisomer or salt thereof, In the formula, R 41 is a cyclic monosaccharide; the wavy line is R b or to said enzyme-cleavable group, 13. The linker compound of claim 12.

14. R 24 and R 25 8. The linker compound of claim 7, wherein is independently a polyhydroxyl selected from cyclic monosaccharides, disaccharides, and polysaccharides.

15. below: or a stereoisomer or salt thereof, In the formula, each R 45 is selected from H and a monosaccharide, a disaccharide, or a polysaccharide; R 46 is selected from cyclic monosaccharides, disaccharides, or polysaccharides; the wavy line is R b or to said enzyme-cleavable group, 15. The linker compound of claim 14.

16. R 24 and R 25 are independently selected from a linear monosaccharide and a substituted linear monosaccharide, wherein the substituted linear monosaccharide is substituted with a monosaccharide, a disaccharide, or a polysaccharide.

17. below: or a stereoisomer or salt thereof, In the formula, R 47 is a linear monosaccharide; each R 49 is selected from monosaccharides, disaccharides, and polysaccharides; the wavy line is R b or to said enzyme-cleavable group, 17. The linker compound of claim 16.

18. R 24 and R 25 are independently selected from linear monosaccharides and substituted monosaccharides, wherein the substituted linear monosaccharides are substituted with one or more substituents selected from carboxyl, ester, and amide, and may be further substituted with a monosaccharide, disaccharide, or polysaccharide.

19. below: or a stereoisomer or salt thereof, In the formula, each R 42 are independently selected from linear monosaccharides and substituted linear monosaccharides; each R 43 are independently selected from hydroxyl, carboxyl, ester, and amide; the wavy line represents R b or to said enzyme-cleavable group, 19. The linker compound of claim 18.

20. R 24 and R 25 is a -C(O)-polyhydroxyl group or a substituted -C(O)-polyhydroxyl group, and R 24 and R 25 and the other is H, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl group, or a substituted polyhydroxyl group; and wherein the substituted -C(O)-polyhydroxyl group and the polyhydroxyl group are substituted with a monosaccharide, a disaccharide, a polysaccharide, a carboxyl, an ester, or an amide.

21. below: or a stereoisomer or salt thereof, In the formula, the wavy line represents R b or to said enzyme-cleavable group, 21. The linker compound of claim 18 or 20.

22. R 24 and R 25 are independently H or substituted -C 1 ~C 8 Alkyl, but R 24 and R 25 and -C cannot both be H; 1 ~C 8 8. The linker compound of claim 7, wherein the alkyl is substituted with hydroxyl and / or carboxyl.

23. below: or a stereoisomer or salt thereof, In the formula, R 48 are H, OH, and CH 2 OH, COOH, or -C substituted with hydroxyl or carboxyl 1 ~C 6 alkyl; the wavy line is R b or to said enzyme-cleavable group, 23. The linker compound of claim 22.

24. R 24 and R 25 One of the groups is H or substituted -C(O)-C 1 ~C 8 alkyl, and R 24 and R 25 The other is a substituted -C(O)-C 1 ~C 8 Alkyl or substituted -C 1 ~C 8 alkyl, where substituted -C(O)-C 1 ~C 8 Alkyl and Substituted-C 1 ~C 8 8. The linker compound of claim 7, wherein the alkyl is substituted with hydroxyl and / or carboxyl.

25. below: or a stereoisomer or salt thereof, In the formula, the wavy line represents R b or to said enzyme-cleavable group, 25. The linker compound of claim 24.

26. R 24 and R 25 is independently selected from H and a chelating agent, wherein the chelating agent is selected from —NR by alkylene, arylene, carbocyclo, heteroarylene, or heterocarbocylo; 24 R 25 may be attached to the nitrogen of R 24 and R 25 8. The linker compound of claim 7, wherein both of are not H.

27. 27. The linker compound of claim 26, wherein the chelator is selected from ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), triethylenetetraminehexaacetic acid (TTHA), benzyl-DTPA, 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA), benzyl-DOTA, 1,4,7-triazacyclononane-N,N',N''-triacetic acid (NOTA), benzyl-NOTA, 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA), and N,N'-dialkyl substituted piperazines.

28. below: or a stereoisomer or salt thereof, In the formula, the wavy line represents R b or to said enzyme-cleavable group, 28. The linker compound of claim 27.

29. Each monosaccharide independently C5 or C6 sugars selected from glucose, ribose, galactose, mannose, arabinose, 2-deoxyglucose, glyceraldehyde, erythrose, threose, xylose, lyxose, allose, altrose, gulose, idose talose, aldose, and ketose; a sugar acid selected from gluconic acid, aldonic acid, uronic acid, and urosonic acid; or an amino sugar selected from glucosamine, N-acetylglucosamine, galactosamine, and N-acetylgalactosamine 21. The linker compound of any one of claims 6 to 20, selected from:

30. 30. The linker compound of any one of claims 1 to 29, wherein the attachment site is formed from a functional group of a 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, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, and protected forms thereof.

31. below: (a) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -R 30 (XXX) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 an alkylene group; R 30 may be substituted C 3 ~C 10 Carbocycle; Thiourea; Optionally substituted thiourea; Urea; optionally substituted ureas; sulfamides; alkyl sulfamides; acylsulfamides; optionally substituted alkyl sulfamides; optionally substituted acylsulfamides; sulfonamides; optionally substituted sulfonamides; guanidines, including alkyl guanidines and aryl guanidines; phosphoramides; or optionally substituted phosphoramides; or R 30 represents azido, alkynyl, substituted alkynyl, -NH-C(O)-alkynyl, -NH-C(O)-alkynyl-R 65 Cyclooctyne; -NH-cyclooctyne, -NH-C(O)-cyclooctyne, or -NH-(cyclooctyne) 2 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (b) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -NH-C(O)-R 31 (XXXI) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 an alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 having R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (c) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -C(O)NH-R 31 (XXXII) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 an alkylene group; R 31 is a branched polyethylene glycol chain, each branch independently having 1 to 26 ethylene glycol subunits and each branch having R at its end 35 having R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, and optionally substituted heteroaryl; n20 is 2 to 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 31 is H or R 22 -NR 24 R 25 and R 21 and R 22 are each independently a bond or C 1 ~C 3 an alkylene group; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 24 and R 25 Both of these cannot be H; n20 is 2 to 26; (e) ~R 20 -R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -N(R 33 -R 31 ) 2 (XXXIV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 an alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 having 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 alkylene-C(O); R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; n20 is 2 to 26; (f) ~R 20 -(R 21 -[CH 2 -CH(OR 34 )-CH 2 -O] n20 -R 36 ) n25 (XXXV) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; Each R 21 are independently a bond, -O- or C 1 ~C 3 an alkylene group; Each R 34 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 Alkylene-NR 24 R 25 and R N is H or C 1 ~C 4 is alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; or a substituted polyhydroxyl group, with the proviso that R 24 and R 25 Both of these cannot be H; Each R 36 are independently H, C 1 ~C 6 Alkylene-C(OH)H-NR 44 R 45 , C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkylene-NR 44 R 45 , -C(O)-NR 24 R 25 , -C(O)N(R N )-C 1 ~C 6 Alkylene-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 are independently H or C 1 ~C 6 is alkyl; R 44 and R 45 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; and a substituted -C(O)-polyhydroxyl group, with the proviso that R 44 and R 45 cannot both be H; each n20 independently is 1 to 26; n25 is 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 , R 22 and R 23 are each independently a bond or C 1 ~C 3 an alkylene group; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 24 and R 25 Both of these cannot be H; each n20 independently is 0 to 26 and each n21 independently is 0 to 26, provided that at least one of n20 or n21 is 2 to 26; n22 is 1 to 5; each n23 is independently 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 an alkylene group; R N is H or C 1 ~C 4 is alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; and a substituted polyhydroxyl group, with the proviso that R 24 and R 25 Both of these cannot be H; Each R 34 are independently H, -[CH 2 -CH(OH)-CH 2 -O] n20 -R 36 , or -C(O)N(R N )-C 1 ~C 6 Alkylene-NR 24 R 25 and Each R 36 are independently H, C 1 ~C 6 Alkylene-C(OH)H-NR 44 R 45 , C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkylene-NR 44 R 45 , -C(O)N(R N )-C 1 ~C 6 Alkylene-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 are independently H or C 1 ~C 6 is alkyl; R 44 and R 45 are each independently selected from H; a polyhydroxyl group; a substituted polyhydroxyl group; a —C(O)-polyhydroxyl group; and a substituted —C(O)-polyhydroxyl group, with the proviso that R 44 and R 45 Both of these cannot be H; n20 is 2 to 26; n21 is 1 to 26; n25 is 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 an alkylene group; R N is H or C 1 ~C 4 is alkyl; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; and a substituted polyhydroxyl group, with the proviso that R 24 and R 25 Both of these cannot be H; n20 is 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 a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 and R 22 are each independently bonded, 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 are independently H or -R 22 -NR 24 R 25 and Each R N are independently H, C 1 ~C 6 Alkyl, or -R 22 -NR 24 R 25 and R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C(O)-C 1 ~C 8 Alkyl; Chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 Both of these cannot be H; each n20 independently is 0 to 26, provided that at least one n20 is 2 to 26; n25 is 1 or 2; or (k) or a stereoisomer or salt thereof, wherein R 20 is part R b or to an enzyme-cleavable group; R 21 , R 22 and R 23 are each independently bonded, 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 are independently H or -R 22 -NR 24 R 25 and Each R N are independently H, C 1 ~C 6 Alkyl, or -R 22 -NR 24 R 25 and R 24 and R 25 are each independently H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C(O)-C 1 ~C 8 Alkyl; Chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from, however, R 24 and R 25 Both of these cannot be H; R 26 is H or C 1 ~C 6 is alkyl; each n20 independently is 0 to 26, provided that at least one n20 is 2 to 26; each n21 independently is 0 to 26, provided that at least one n21 is 2 to 26; 7. The linker compound of any one of claims 1 to 6, comprising a polar group having a formula selected from:

32. 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) or a stereoisomer or salt thereof, During the ceremony, R 20 is part R b is a linking group to or to said enzyme-cleavable group; R 21 and R 22 are each independently a bond or C 1 ~C 3 an alkylene group; R 31 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 35 having 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 alkylene-C(O); R 35 are azide, alkynyl, and alkynyl-R 65 , cyclooctyne, or cyclooctyne-R 65 where R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; the wavy (~) line is R 20 n20 indicates a binding site for The linker compound of any one of claims 1 to 6.

33. below: and a polar group formed from a precursor group selected from In the formula, R 65 is selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocycle, optionally substituted aryl, optionally substituted heterocarbocycle, or optionally substituted heteroaryl; the wavy line represents the moiety R b or to said enzyme-cleavable group, 33. The linker compound of claim 31 or 32.

34. R b 34. The linker compound of any one of claims 31 to 33, wherein the attachment site to or to the enzyme-cleavable group is formed from a functional group of a 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, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, and protected forms thereof.

35. 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 stereoisomer or salt thereof, During the ceremony, R 20 is part R b is a linking group to or to said enzyme-cleavable group; R 41 and R 42 are each independently a bond or C 1 ~C 6 alkylene; Each R 43 are independently bonded, C 1 ~C 12 Alkylene, -OC 1 ~C 12 Alkylene, -C(=O)-, -NR a -C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkylene-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 Alkylene-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 wherein each alkylene is selected from the group consisting of hydroxyl, SO 3 may be substituted with H and / or oxo, R a is H, C 1 ~C 6 alkyl, polyhydroxyl, or substituted polyhydroxyl group, and R 46 and R 47 One of the two is H or C 1 ~C 12 alkylene and the other is C 1 ~C 12 alkylene, wherein the C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom. 44 R 45 is bound to; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; n40 is 2 to 26, with the proviso that R 44 and R 45 Both of these cannot be H; n40 is 2 to 26; n41 is 1 to 6; n42 is 1 to 6, The linker compound of any one of claims 1 to 6.

36. formula: ~R 20 -(R 41 -[O-CH 2 -CH 2 ] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLI) or a stereoisomer or salt thereof, During the ceremony, R 20 is part R b is a linking group to or to said enzyme-cleavable group; R 41 and R 42 are each independently a bond or C 1 ~C 6 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 Alkylene-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 Alkylene-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 wherein each alkylene is selected from the group consisting of hydroxyl, SO 3 may be substituted with H and / or oxo, R a is H, C 1 ~C 6 alkyl, polyhydroxyl, or substituted polyhydroxyl group, and R 46 and R 47 One of the two is H or C 1 ~C 12 alkylene and the other is C 1 ~C 12 alkylene, wherein the C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom. 44 R 45 is bound to; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 Both of these cannot be H; n40 is 1 to 26; n41 is 1 to 6; n42 is 1 to 6, The linker compound of any one of claims 1 to 6.

37. formula: ~R 20 -(R 41 -[O-CH 2 -CH 2 ] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLII) or a stereoisomer or salt thereof, During the ceremony, R 20 is part R b is a linking group to or to said enzyme-cleavable group; R 41 and R 42 are each independently a bond or C 1 ~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 Alkylene-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 Alkylene-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 wherein each alkylene is selected from the group consisting of hydroxyl, SO 3 may be substituted with H and / or oxo, R a is H, C 1 ~C 6 alkyl, polyhydroxyl, or substituted polyhydroxyl group, and R 46 and R 47 One of the two is H or C 1 ~C 6 alkylene and the other is C 1 ~C 12 alkylene, wherein the C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom. 44 R 45 is bound to; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 Both of them cannot be H; n40 is 1 to 16; n41 is 1 to 4; n42 is 1 to 4, The linker compound of any one of claims 1 to 6.

38. R 20 is formed from a functional group of a precursor compound of said polar group, said functional group being selected from halo, aldehyde, carboxyl, amino, alkynyl, azide, hydroxyl, carbonyl, carbamate, thiol, urea, thiocarbamate, thiourea, sulfonamide, acylsulfonamide, alkyl sulfonate, triazole, azadibenzocyclooctyne, hydrazine, carbonylalkylheteroaryl, or a protected form thereof.

39. R 20 but has the following structure: or a stereoisomer thereof, In the formula, R is H, C 1 ~C 6 an alkyl or polyhydroxyl group, n is 0 to 12; is R b indicates the site of attachment to or to said enzyme-cleavable group, indicates the site of attachment of the polar group to the remainder of the group; 38. The linker compound of any one of claims 7, 31, 32, and 35-37.

40. R 20 but has the following structure: or a stereoisomer thereof, wherein n is 0 to 12; is R b indicates the site of attachment to or to said enzyme-cleavable group, indicates the site of attachment of the polar group to the remainder of the group; 38. The compound of any one of claims 7, 31, 32, and 35-37.

41. R 43 -(NR 44 R 45 ) n41 but has the following structure: or a stereoisomer thereof, In the formula, R a is H, C 1 ~C 6 alkyl, polyhydroxyl, or substituted polyhydroxyl; p is an integer from 1 to 6; R to the remainder of the polar group 43 indicates the binding site of 41. The linker compound of any one of claims 35 to 40.

42. R 43 -(NR 44 R 45 ) n41 but has the following structure: or a stereoisomer thereof, During the ceremony, R to the remainder of the polar group 43 indicates the binding site of 41. The linker compound of any one of claims 35 to 40.

43. -NR 44 R 45 but has the following structure: or a stereoisomer thereof, During the ceremony, -NR to the remainder of the polar group 44 R 45 indicates the binding site of 43. The linker compound of any one of claims 35 to 42.

44. Prior to attachment to the linker unit, the structure: and a polar group having one of During the ceremony, (*) indicates part R b indicates the site of attachment to or to said enzyme-cleavable group; Each R is independently H or C 1 ~C 6 is alkyl; R' is H, C 1 ~C 6 Alkyl, -N(R 24 )(R 25 ), or -CO 2 H; each n is independently 1 to 12; X is O, NR, or -CH 2 - and; V is a bond or C 1 ~C 6 is alkyl; R 24 and R 25 one of which is 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; and R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 Selected from: R 24 and R 25 The other is H; a polyhydroxyl group; a substituted polyhydroxyl group; a -C(O)-polyhydroxyl group; a substituted -C(O)-polyhydroxyl group; a substituted -C(O)-C 1 ~C 8 Alkyl; Chelating agent; R 28 is a sugar unit of formula (XII) or (XIII) -C(O)-R 28 and polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, with the proviso that R 24 and R 25 Both of them cannot be H.

45. The linker compound of any one of claims 1 to 44.

45. below: (a) ~R 40 -(R 43 -R 41 -[O-CH 2 -CH 2 ] n40 -R 46 -[O-CH 2 -CH 2 ] n40 -R 42 -R 43 -(NR 44 R 45 ) n41 ) n42 (XLIII) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 41 and R 42 are each independently a bond or C 1 ~C 6 alkylene; Each R 43 are independently bonded, C 1 ~C 12 Alkylene, -OC 1 ~C 12 Alkylene, -C(=O)-, -NH-C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkylene-NH-, -C(O)-C 1 ~C 12 Alkylene, -C 1 ~C 12 Alkylene-C(O)-, -NH-C 1 ~C 12 Alkylene-C(O)-, -C(O)-C 1 ~C 12 Alkylene-NH-, -NH-C(O)-NH-, -NH-C(O)-, -NH-C(O)-C 1 ~C 12 Alkylene, -C(O)-NH-C 1 ~C 12 Alkylene, C 1 ~C 12 Alkylene-NH-C(O)-, -heteroarylene, heteroaryl-C 1 ~C 12 Alkylene, Heteroaryl-C 1 ~C 12 Alkylene -C(O)- or -C(O)NR 46 R 47 where R 46 and R 47 One of the two is H or C 1 ~C 12 alkylene and the other is C 1 ~C 12 alkylene; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 Both of these cannot be H; Each R 46 is independently -NR 50 -, -NR 50 -C 1 ~C 6 Alkylene-NR 50 -, -NR 50 -C(O)-NR 50 -S(O) 2 -NR 50 -, or -NR 50 -C(O)-C 1～6 alkylene-; Each R 50 are independently H, C 1 ~C 6 alkyl, or polyhydroxyl groups; each n40 is independently 2 to 26; n41 is 1 to 6; n42 is 1-6; (b) ~R 40 -(R 51 -[O-CH 2 -CH 2 ] n43 -R 52 -X 1 -R 55 -X 2 -R 53 -[O-CH 2 -CH 2 ] n43 -R 54 -[X 3 -R 56 ] n44 -R 57 ) n45 (XLIV) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 51 , R 52 , R 53 and R 54 are each independently a bond or C 1 ~C 6 alkylene; X 1 , X 2 and X 3 are each independently -NR N -C(O)- or -C(O)-NR N - and; Each R N are independently H, C 1 ~C 6 represents an alkyl, or polyhydroxyl group; R 55 and R 56 each independently represents a divalent polyhydroxyl group; R 57 is H, OH or C 1 ~C 6 is alkyl; each n43 is independently 0 to 26, provided that at least one n43 is 1 to 26; n44 is 0 to 10; n45 is 1 or 2; or (c) ~R 40 -R 51 -[O-CH 2 -CH 2 ] n43 -R 52 -N-(R 53 -X 1 -R 54 -[O-CH 2 -CH 2 ] n43 -(NR 44 R 45 )) 2 (XLV) or a stereoisomer or salt thereof, wherein R 40 is part R b or to an enzyme-cleavable group; R 51 , R 53 and R 54 each independently optionally bonded or substituted C 1 ~C 6 alkylene; R 52 is a bond, C 1 ~C 6 alkylene, —C(O)—, or —OC(O)—; each X 1 is independently -NR N -C(O)- or -C(O)-NR N - and; Each R N are independently H, C 1 ~C 6 represents an alkyl, or polyhydroxyl group; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 Both of these cannot be H; each n43 is independently 2 to 26; 7. The linker compound of any one of claims 1 to 6, comprising a polar group having a formula selected from:

46. The following structure prior to attachment to the enzyme-cleavable group and / or to the linker unit: and a polar group having one of During the ceremony, (*) indicates part R b indicates the site of attachment to or to said enzyme-cleavable group; each R is independently H, an alkyl, or a polyhydroxyl group; R 44 and R 45 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, or a substituted -C(O)-polyhydroxyl group, where the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, with the proviso that R 44 and R 45 Both of these cannot be H; each n is independently 1 to 12; 46. ​​The linker compound of claim 45.

47. A polar group having a formula selected from the following, or a stereoisomer or salt thereof: During the ceremony, Each Y is independently R 76 or and Each R 76 are independently H, acetyl, -P(=O)(OH) 2 , or -(CH 2 ) v -OS(=O) 2 (OH); Each R a and R b are independently H or R a and R b together with the carbon to which they are attached form an oxo group; each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; each v is independently 1 to 6; Each * is R b or to said enzyme-cleavable group, The linker compound of any one of claims 1 to 6.

48. A polar group having a formula selected from the following, or a stereoisomer or salt thereof: During the ceremony, Each R 76 are independently H, acetyl, -P(=O)(OH) 2 , or -(CH 2 ) v S(=O) 2 (OH); each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; each v is independently 1 to 6; Each * is R b or to said enzyme-cleavable group, The linker compound of any one of claims 1 to 6.

49. A polar group having a formula selected from the formula: During the ceremony, each q is independently 2 to 26; each m is independently 1 to 4; each n is independently 1 to 4; Each * is R b or to said enzyme-cleavable group, The linker compound of any one of claims 1 to 6.

50. Y is R 76 48. The linker compound of claim 47, wherein:

51. Y is 48. The linker compound of claim 47, wherein:

52. Each R a and R b 48. The linker compound of claim 47, wherein:

53. R a and R b 48. The linker compound of claim 47, wherein, together with the carbon to which they are attached, form an oxo group.

54. 48. The linker compound of any one of claims 45 to 47, wherein q is 10 to 20.

55. 50. The linker compound of any one of claims 57 to 49, wherein q is 12.

56. below: or a stereoisomer or salt thereof, During the ceremony, each Z is joined at *, more independently selected, each is R b or to said enzyme-cleavable group, 50. The linker compound of any one of claims 1-6, 35-37, or 47-49.

57. The polar group has the formula: or a stereoisomer or salt thereof, During the ceremony, (a) L 70 is C 1 ~C 8 Alkylene, C 1 ~C 8 Alkylene-C(O)-, -C(O)-C 1 ~C 8 Alkylene-, and -C(O)-C 1 ~C 8 alkylene-C(O)-, * is R b , to an enzyme-cleavable group, or to the remainder of a polar group; R 70 Ha~NR 71 (R 72 -R 73 ), where R 71 is H, C 1 ~C 12 Alkyl, substituted C 1 ~C 12 alkyl, or polyethylene glycol (which may have 1 to 12 ethylene glycol subunits); R 72 is a bond or an optionally substituted C 1 ~C 3 is selected from alkylene, optionally substituted ether, optionally substituted thioether, optionally substituted ketone, optionally substituted amide, polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), optionally substituted carbocycle, optionally substituted aryl, or optionally substituted heteroaryl; R 73 is a carboxyl or polycarboxyl, wherein the polycarboxyl contains 1 to 10, or 1 to 6, or 1 to 4 carboxyl groups, and the carboxyl groups are interconnected by alkyl, alkylene, substituted alkyl, substituted alkylene, heteroalkyl, heteroalkylene, amino, and / or amido; or (b) L 70 is C 1 ~C 8 Alkylene, C 1 ~C 8 Alkylene-C(O)-, -C(O)-C 1 ~C 8 Alkylene-, and -C(O)-C 1 ~C 8 alkylene-C(O)-, * is R b , to an enzyme-cleavable group, or to the remainder of a polar group; R 70 Ha~NR 71 (R 75 - (R 73 ) 2 ), where R 71 is H, C 1 ~C 12 Alkyl, substituted C 1 ~C 12 alkyl, or polyethylene glycol (which may have 1 to 12 ethylene glycol subunits); R 75 C is an optionally branched substituted 1 ~C 3 alkylene, optionally substituted ether, optionally substituted thioether, optionally substituted ketone, optionally substituted amide, polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), optionally substituted carbocycle, optionally substituted aryl, or optionally substituted heteroaryl, and each R 73 is independently a carboxyl or polycarboxyl, wherein the polycarboxyl contains 1 to 10, or 1 to 6, or 1 to 4 carboxyl groups, and the carboxyl groups are interconnected by alkyl, alkylene, substituted alkyl, substituted alkylene, heteroalkyl, heteroalkylene, amino, and / or amido; or (c) L 70 is C 1 ~C 8 Alkylene, C 1 ~C 8 Alkylene-C(O)-, -C(O)-C 1 ~C 8 Alkylene-, and -C(O)-C 1 ~C 8 alkylene-C(O)-, * is R b , to an enzyme-cleavable group, or to the remainder of a polar group; R 70 Ha~N(R 74 -R 73 )(R 72 - R 73 ), where R 72 and R 74 each independently represents an optionally substituted C 1 ~C 3 is selected from alkylene, optionally substituted ether, optionally substituted thioether, optionally substituted ketone, optionally substituted amide, polyethylene glycol (optionally having 1 to 12 ethylene glycol subunits), optionally substituted carbocycle, optionally substituted aryl, or optionally substituted heteroaryl; 73 are independently carboxyl or polycarboxyl, wherein the polycarboxyl contains 1 to 10, or 1 to 6, or 1 to 4 carboxyl groups, and the carboxyl groups are interconnected by alkyl, alkylene, substituted alkyl, substituted alkylene, heteroalkyl, heteroalkylene, amino, and / or amido; The linker compound of any one of claims 1 to 6.

58. 58. The linker compound of any one of claims 1 to 57, comprising a polar group comprising said polymer unit and a saccharide unit.

59. 58. The linker compound of any one of claims 1-57, comprising a polar group comprising at least two polymer units.

60. 58. The linker compound of any one of claims 1 to 57, comprising a polar group comprising said polymer unit and a carboxyl unit.

61. 58. The linker compound of any one of claims 1 to 57, comprising at least two polar groups.

62. 58. The linker compound of any one of claims 1 to 57, comprising a polar group comprising said polymer unit, said sugar unit, and said carboxyl unit.

63. 58. The linker compound of any one of claims 1-57, comprising a polar group comprising at least two polymer units, at least one sugar unit, and at least one carboxyl unit.

64. 58. The linker compound of any one of claims 1 to 57, wherein said enzyme-cleavable group comprises at least two amino acid units.

65. 65. The linker compound of any one of claims 1 to 64, comprising at least one said polar group attached to said enzyme-cleavable group.

66. The following structure: and During the ceremony, R C is a bond or C 1～6 alkylene; The wavy line on the amino group indicates the attachment site for a Stretcher group or indicates an H prior to attachment to the Stretcher group; β- is the attachment site to the at least one polar group; the benzyl H on the benzyl OH may be replaced with a bond to at least one of the Drug units or a bond to a linking group attached to at least one of the Drug units; 65. The linker compound of any one of claims 1 to 64.

67. 67. The linker compound of any one of claims 1 to 66, wherein said enzyme-cleavable group comprises a peptide that is cleavable by an intracellular protease.

68. 68. The linker compound of claim 67, wherein the intracellular protease is cathepsin B.

69. 69. The linker compound of claim 68, wherein the enzyme-cleavable group comprises a cleavable peptide comprising a valine-citrulline peptide, a valine-alanine peptide, a valine-lysine peptide, a phenylalanine-lysine peptide, or a glycine-glycine-phenylalanine-glycine peptide.

70. The following structure: including one of the During the ceremony, R C is a bond or C 1～6 alkylene; A wavy line on an amino group indicates the attachment site for the Stretcher group, or indicates an H prior to attachment to the Stretcher group; β- is the attachment site to the at least one polar group; the H on the benzyl OH may be replaced with a bond to at least one of the Drug units or a bond to a binding site for at least one of the Drug units; 70. The linker compound of any one of claims 67 to 69.

71. The following structure: and wherein the wavy line on the amino group indicates the site of attachment to the Stretcher group or represents an H prior to attachment to the Stretcher group; and the H on the benzyl OH may be replaced with a bond to at least one of the Drug units or a bond to a linking group attached to at least one of the Drug units. The linker compound of any one of claims 1 to 5.

72. 70. The linker compound of any one of claims 1-69, wherein said enzyme-cleavable group is attached to said Stretcher group by a non-peptide linking group.

73. The non-peptide linking group is optionally substituted C 1 ~C 10 Alkylene, optionally substituted C 2 ~C 10 Alkenylene, optionally substituted C 2 ~C 10 73. The linker compound of claim 72, selected from alkynylene, or optionally substituted polyethylene glycol.

74. 74. The linker compound of any one of claims 1 to 73, comprising said Stretcher group attached to said enzyme-cleavable group.

75. The stretcher group may be: wherein R 17 -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 1 to 26), -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C 1 ~C 8 alkylene- (where b is 1 to 26), -arylene-, -C 1 ~C 10 Alkylene-arylene-, -arylene-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)- (wherein n is 1 to 26), C 1 ~C 10 Heteroalkylene-C(=O)-, -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C(=O)- (wherein b is 1 to 26), -(CH 2 -O-CH 2 ) b -C 1 ~C 8 alkylene-C(=O)- (wherein b is 1 to 26), -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C 1 ~C 8 alkylene-C(=O)- (wherein 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-arylene-C(=O)-, -arylene-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- (wherein b is 1 to 26), -(CH 2 -O-CH 2 ) b -C 1 ~C 8 alkylene-NH- (wherein b is 1 to 26), -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C 1 ~C 8 alkylene-NH- (wherein b is 1 to 26), -C 1 ~C 8 Alkylene-(C(=O))-NH-(CH 2 -O-CH 2 ) b -C(=O)- (wherein b is 1 to 26), -C 1 ~C 8 Alkylene-(C(=O))-NH-(CH 2 -O-CH 2 ) b -C 1 ~C 8 alkylene-C(=O)- (wherein b is 1 to 26), -C 1 ~C 8 Alkylene-NH-(C(=O))-(CH 2 -O-CH 2 ) b -NH- (wherein b is 1 to 26), -C 1 ~C 8 Alkylene-NH-(C(=O))-(CH 2 -O-CH 2 ) b -C 1 ~C 8 alkylene-NH- (wherein 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 Alkylene-arylene-S-, -arylene-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 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 alkylene)(CH 2 OCH 2 ) p2 C(O)-, or their open ring forms, wherein p2 is 1 to 26; * is the site of attachment to the targeting group and the wavy line is the site of attachment to the enzyme-cleavable group; 75. The linker compound of claim 74.

76. The stretcher group is: Selected from In the formula, wavy line indicates the attachment site of the Stretcher group to the enzyme-cleavable group, and the attachment site to the targeting group is on a maleimide, a primary amine, or an alkyne functional group; 75. The linker compound of claim 74.

77. The following structure: and wherein the H on the benzyl OH may be replaced with a bond to at least one of the Drug units or a bond to a linking group attached to at least one of the Drug units. The linker compound of claim 1.

78. 78. The linker compound of any one of claims 1 to 77, wherein said linker compound is attached at said attachment site to at least one Drug unit or to a linking group attached to at least one Drug unit.

1. A drug-linker compound comprising:

79. 79. The Drug-Linker of Claim 78, wherein said Drug unit is selected from a cytotoxic agent, an immunomodulator, a nucleic acid, a growth inhibitor, a PROTAC, a toxin, a radioisotope, and a chelating ligand.

80. 80. The Drug-Linker of Claim 79, wherein the Drug unit is a cytotoxic agent.

81. 81. The drug-linker of claim 80, wherein the cytotoxic agent is selected from the group consisting of an auristatin, a maytansinoid, a camptothecin, a duocarmycin, and a calicheamicin.

82. 82. The drug-linker of claim 81, wherein the cytotoxic agent is an auristatin.

83. 83. The drug-linker of claim 82, wherein the cytotoxic agent is MMAE or MMAF.

84. 82. The drug-linker of claim 81, wherein the cytotoxic agent is camptothecin.

85. 85. The drug-linker of claim 84, wherein the cytotoxic agent is exatecan, or SN-38, or DxD.

86. 86. The drug-linker of claim 85, wherein the cytotoxic agent is RS-exatecan or SS-exatecan.

87. 82. The drug-linker of claim 81, wherein the cytotoxic agent is calicheamicin.

88. 82. The drug-linker of claim 81, wherein the cytotoxic agent is a maytansinoid.

89. 89. The drug-linker of claim 88, wherein the maytansinoid is maytansine, maytansinol, or ansamatocin-2.

90. 80. The Drug-Linker of Claim 79, wherein the Drug unit is an immunomodulator.

91. 91. The drug-linker of claim 90, wherein the immunomodulator is selected from a TRL7 agonist, a TLR8 agonist, a STING agonist, or a RIG-I agonist.

92. 92. The drug-linker of claim 91, wherein the immunomodulator is a TLR7 agonist.

93. The drug-linker of claim 92, wherein the TLR7 agonist is an imidazoquinoline, imidazoquinoline amine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzimidazol-2-amine, tetrahydropyridopyrimidine, heteroarothiadiazide-2,2-dioxide, benzonaphthyridine, guanosine analog, adenosine analog, thymidine homopolymer, ssRNA, CpG-A, polyG10, or polyG3.

94. 92. The drug-linker of claim 91, wherein the immunomodulator is a TLR8 agonist.

95. 95. The drug-linker of claim 94, wherein the TLR8 agonist is selected from imidazoquinolines, thiazoloquinolines, aminoquinolines, aminoquinazolines, pyrido[3,2-d]pyrimidine-2,4-diamines, pyrimidine-2,4-diamines, 2-aminoimidazoles, 1-alkyl-1H-benzimidazol-2-amines, tetrahydropyridopyrimidines, or ssRNA.

96. 92. The drug-linker of claim 91, wherein the immunomodulator is a STING agonist.

97. 92. The drug-linker of claim 91, wherein the immunomodulator is a RIG-I agonist.

98. The drug-linker of claim 97, wherein the RIG-I agonist is selected from KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400, and KIN2000.

99. 80. The Drug-Linker of Claim 79, wherein said Drug unit is a chelating ligand.

100. 100. The drug-linker of claim 99, wherein the chelating ligand is selected from platinum (Pt), ruthenium (Ru), rhodium (Rh), gold (Au), silver (Ag), copper (Cu), molybdenum (Mo), titanium (Ti), or iridium (Ir); a radioisotope, 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.

101. The following structure:

79. The drug-linker of claim 78, having one of:

102. 102. A conjugate comprising a targeting group attached to the drug-linker of any one of claims 78-101, wherein the targeting group specifically binds to a target molecule.

103. 103. The conjugate of claim 102, wherein the targeting group is selected from an antibody or an antigen-binding portion thereof.

104. 104. The conjugate of claim 103, wherein the targeting group is a monoclonal antibody, Fab, Fab', F(ab'), Fv, disulfide-linked Fc, scFv, single domain antibody, diabody, bispecific antibody, or multispecific antibody.

105. 103. The conjugate of claim 102, wherein said targeting group is a diabody, DART, anticalin, affibody, avimer, DARPin, or adnectin.

106. 106. The conjugate of any one of claims 102 to 105, wherein said targeting group is monospecific.

107. 107. The conjugate of any one of claims 102 to 106, wherein the targeting group is divalent.

108. 106. The conjugate of any one of claims 102 to 105, wherein said targeting group is bispecific.

109. The average drug loading of the complex (p load 109. The conjugate of any one of claims 102-108, wherein n is from about 1 to about 8, about 2, about 4, about 6, about 8, about 10, about 12, about 14, about 16, about 3 to about 5, about 6 to about 8, or about 8 to about 16.

110. below: Selected from where Ab is a targeting group and n is p load That is, 110. The conjugate of any one of claims 102 to 109.

111. The conjugate of any one of claims 102 to 110, wherein the target molecule is CD19, CD20, CD30, CD33, CD70, LIV-1, EGFRv3, or HER2.

112. The conjugate of any one of claims 102 to 111, wherein the target molecule is a cancer-associated antigen.

113. The target molecule is CD19, CD20, CD30, CD33, CD38, CA125, HER2, MUC-1, prostate-specific membrane antigen (PSMA), CD44 surface adhesion molecule, mesothelin (MLSN), carcinoembryonic antigen (CEA), epidermal growth factor receptor (EGFR), EGFRvIII, vascular endothelial growth factor receptor 2 (VEGFR2), high molecular weight melanoma-associated antigen (HMW-MAA), MAGE-A1, IL-13R-a2, GD2, 1p19q, HER2, ABL 1, AKT1, ALK, APC, AR, ATM, BRAF, BRCA1, BRCA2, cKIT, cMET, CSF1R, CTNNB1, FGFR1, FGFR2, FLT3, GNA11, GNAQ, GNAS, HRAS, IDH1 , IDH2, JAK2, KDR(VEGFR2), KRAS, MGMT, MGMT-Me, MLH1, MPL, NOTCH1, NRAS, PDGFRA, Pgp, PIK3CA, PR, PTEN, RET, RRM1, SMO, SPA RC, TLE3, TOP2A, TOPO1, TP53, TS, TUBB3, VHL, CDH1, ERBB4, FBXW7, HNF1A, JAK3, NPM1, PTPN11, RB1, SMAD4, SMARCB1, STK1, MLH 1, MSH2, MSH6, PMS2, ROS1, ERCC1, 5T4(TPBG), B7-H3, CCR7, CD105, CD22, CD46, CD47, CD56, CD70, CD71, CD79b, CDH6, CLDN6, CL The conjugate of any one of claims 102 to 111, which is DN18.2, CLEC12A, DLL3, DR5, ERBB3 (HER3), EPCAM, FOLR1, IGF1R, IL2RA (CD25), IL3RA, ITGB6, LIV-1, LRRC15, mesothelin (MSLN), NaPi2b (SLC34A2), nectin-4, PTK7, ROR1, SEZ6, SLC44A4, SLITRK6, tissue factor (TF), TROP2, or B7-H4.

114. The targeting group is selected from the group consisting of rituximab (Rituxan®), trastuzumab (Herceptin®), pertuzumab (Perjeta®), bevacizumab (Avastin®), ranibizumab (Lucentis®), cetuximab (Erbitux®), alemtuzumab (Campath®), panitumumab (Vectibix®), ibritumomab 105. The conjugate of any one of claims 102-104, which is an antibody comprising tiusetan (Zevalin®), tositumomab (Vexar®), ipilimumab, zalutumumab, dalotuzumab, figitumumab, ramucirumab, galiximab, faretuzumab, ocrelizumab, ofatumumab (Arzera®), tositumumab, ibritumomab, CD20 antibody 2F2 (HuMax-CD20), 7D8, IgM2C6, IgG1 2C6, 11B8, B1, 2H7, LT20, 1FS, or AT80, daclizumab (Zenapax®), or an anti-LHRH receptor antibody comprising clone A9E4, F1G4, AT2G7, GNRH03, or GNRHR2, or a fragment thereof.

115. 115. A pharmaceutical composition comprising the conjugate of any one of claims 102 to 114 and a pharmaceutically acceptable carrier.

116. 1. A method of treating a subject in need thereof, comprising: administering to said subject a conjugate of any one of claims 102 to 114 or a pharmaceutical composition of claim 115; the subject has cancer or an autoimmune disease, and the complex binds to a target antigen associated with the cancer or autoimmune disease; The method.

117. Structure of Formula (A): or a salt thereof, During the ceremony, (i) β is R 20 (-R 21 -[O-CH 2 -CH 2 ] n20 -R 22 -NR 24 R 25 ) z where: R 20 is NH, and R 21 and R 22 are each independently a bond or C 1 ~C 3 alkylene; R 24 and R 25 are each independently selected from H; a polyhydroxyl group; and a —C(O)-polyhydroxyl group; provided that R 24 and R 25 cannot both be H; z is 1 or 2; n20 is 2 to 26; (ii)R a is H or C 1 ~C 6 is alkyl; (iii)R c is a bond, -C(O)-, -S(O)-, -SO 2 -, C 1～6 Alkylene, C 1～6 Alkynylene, or C 1～6 alkynylene-triazolyl; (iv)R 1 is a bond, -C(O)-, or C 1～6 alkylene; (v) δ is selected from drugs; (vi) α is where: R 2 is a peptide having 2 to 5 amino acids; R 3 -C 1 ~C 10 Alkylene-C(=O)-, -C 1 ~C 10 Alkylene-, -C 1 ~C 10 Alkylene-C(O)NH-C 1 ~C 8 Alkylene-[O-CH 2 -CH 2 ] n -C(O)- (where n is 1 to 26), or -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C(=O)- (wherein b is 1 to 26); Drug-linker compounds.

118. R 20 but 118. The drug-linker compound of claim 117, wherein:

119. β is and n20 is 4 to 12.

120. β is 118. The drug-linker compound of claim 117, wherein:

121. R 2 but 121. The drug-linker compound of any one of claims 117-120, selected from:

122. R 2 but 122. The drug-linker compound of any one of claims 117-121, selected from:

123. R 3 but 123. The drug-linker compound of any one of claims 117-122, selected from:

124. R 3 but 124. The drug-linker compound of any one of claims 117-123, selected from:

125. R c The drug-linker compound of any one of claims 117-124, wherein is -C(O)-.

126. R 1 The drug-linker compound of any one of claims 117-125, wherein is -C(O)-.

127. 127. The Drug-Linker compound of any one of claims 117-126, wherein the drug is selected from a cytotoxic agent.

128. The drug-linker compound of any one of claims 117-126, wherein the drug is MMAE or MMAF.

129. The drug-linker compound of any one of claims 117-126, wherein the drug is MMAE.

130. The drug-linker compound of any one of claims 117-126, wherein the drug is exatecan.

131. 127. The Drug-Linker compound of any one of claims 117-126, wherein the drug is selected from immunomodulators.

132. δ is 127. The drug-linker compound of any one of claims 117-126, selected from:

133. δ is The drug-linker compound of any one of claims 117 to 126, wherein

134. δ is The drug-linker compound of any one of claims 117-126, wherein the compound is

135. 135. A conjugate comprising a targeting group attached to the drug-linker of any one of claims 117-134, wherein the targeting group specifically binds to a target molecule.

136. 136. The conjugate of claim 135, wherein the ratio of drug-linker to targeting group is represented by a DAR value, and said DAR value is from 1 to 8.

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