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

Abstract

Aspects of the present invention provide polar groups, linker compounds, linker drugs, and conjugates thereof. TIFF2026503087000657.tif153145

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to International Application No. PCT / CN2023 / 071781, 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 agent (i.e., the number of drugs attached to each targeting agent (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] 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, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 6-([O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 ) n3 (Ia) or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; Each R 3 are independently bonded, C1 to 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)-, -C1-C 12 Alkylene-NR a -C(O)-, -C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -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)-, -NR a -C(O)-C1~C 12Alkylene-C(O)-, -C(O)-NR a -C1~C 12 Alkylene-(CH(OH)) 1~8 -C1~C 12 Alkylene-, -O-CH2-CH2, -OC(O)-NR a -C1~C 12 Alkylene, -O-CH2-CH(OH)-C(O)-, -O-CH2-CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C1-C 12 Alkylene, C1-C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-C1-C 12 Alkylene-NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-NR a -C1~C 12 Alkylene-, -[C(O)-(CH2) 1~8 -NR a ] 1~8 -, Triazolyl, -C1~C 12 Alkylene-triazolyl-, -N(polyhydroxyl group)-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, and each R a are independently H, C 1~6 alkyl, wherein any of the above alkylene groups may be substituted with —SO3H; Each R 4and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R 6 teeth, (i) TIFF2026503087000002.tif26128In the above formula, each n 3 and n 4 are independently between 0 and 1, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; m is 1 to 4; each v is independently 1 to 6; n 2 is 1; (ii) TIFF2026503087000003.tif19128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, n 6 is 1 to 10, each p is independently 0 to 6; n 2 is 1; (iii) TIFF2026503087000004.tif25128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; q is 1 to 8; each v is independently 1 to 6; n 2 is 1; (iv) TIFF2026503087000005.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, each p is independently 0 to 6; n 2 is 1; (v) -R 10 -[O-CH2-CH2] 1~8 -R 10 - In the above formula, Each R b are independently H or C 1~6 is alkyl, Each R 10 is independent TIFF2026503087000006.tif24128, each p is independently 1 to 6; Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), q is 1 to 8; n 2 is 1; and (vi) -N-(R1 -XR 2 -)2 In the above formula, Each X is independently -NR a -C(O)- or -C(O)NR a - and n 2 is 2; More selected; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; each n 0 are independently 2 to 26; each n 1 are independently 1 to 6; n 3 is 1 to 6, polar group A linker compound is provided, comprising:

[0008] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 2 -(NR 4 R 5 ) n1 ) n3 (Ia') or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; Each R 3are independently -N(polyhydroxyl group), triazolyl, -C1 to -C 12 alkylene-triazolyl-, TIFF2026503087000007.tif101128; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R a are independently H or C 1~6 is alkyl; TIFF2026503087000008.tif8128 is R 0 to R 3 indicates the binding site of; Wavy line TIFF2026503087000009.tif8128 is R 1 to R 3 indicates the binding site of; each p is 1 to 6; each n 0 are independently 2 to 8; each n 1 are independently 1 to 6; n 3 is 1 to 6, polar group A linker compound is provided, comprising:

[0009] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 1 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n3 (Ia") or a stereoisomer or salt thereof, wherein: (i) R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; R 3 is -C(O)-; R 4 is H; R 5 is independently a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; n 0 are independently 2 to 26; n 1 is 1 to 6; n 3 is 1-6; (ii) R 0 is -C(O)-; R 1 , R 2 , and R 3 are bonds respectively; R4 and R 5 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; n 0 is 6; n 1 is 1 to 6; n 3 is 1; (iii) R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C6 alkylene; R 3 Ha-NR a -C(O)-C1~C 12 alkylene-C(O)-, where the alkylene is substituted with -SO3H; R a is H or C 1~6 is alkyl; R 4 and R 5 are each independently H, a carboxyl-containing moiety, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; The wavy line (~) is R 0indicates the binding site of the amino acid unit to; each n 0 are independently 1 to 26; n 1 is 1 to 6; n 3 is 1 to 6; or (iv) R 0 teeth TIFF2026503087000010.tif25128; Each R 1 are independently a bond or C1-C6 alkylene; R 2 and R 3 are bonds respectively; R 4 and R 5 are each independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R a are independently H or C 1~6 is alkyl; Wavy line TIFF2026503087000011.tif7128 is the R to the rest of the polymer units 0 indicates the binding site of; Wavy line (~*) is R 0 indicates the binding site of the amino acid unit to; n 0 is 1 to 8; n 1 is 1 to 6; n 3 is 2, polar group A linker compound is provided, comprising:

[0010] In some embodiments, the following: (a) A linker unit having 1 to 4 attachment sites per drug unit, wherein the linker unit has the formula: TIFF2026503087000012.tif49128 or a stereoisomer or salt thereof, wherein: α- represents the direct or indirect attachment site to the amino acid unit; δ- represents an attachment site to at least one of the Drug units or for a linking group attached to at least one of the Drug units; R a is H or C 1~6 is alkyl, Linker units; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to an amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit; A linker compound is provided, comprising:

[0011] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to an amino acid unit, the polymer unit comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit being (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, Polyamides containing TIFF2026503087000013.tif19128, or stereoisomers thereof; (ii) Each R b are independently H or C 1~6 is alkyl, and n 0 are independently 2 to 26, a polyether containing TIFF2026503087000014.tif22128, or a stereoisomer thereof; or (iii) any combination thereof Including, polar group A linker compound is provided, comprising:

[0012] In some embodiments, at least one polar group attached to an amino acid unit has the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 6 -([O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 ) n3 (Ia), ~R 0 -(R 3 -R 1 -[O-CH2-CH(OH)-CH2] n0 -R 6 -[O-CH2-CH(OH)-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n3 (Ib), or A linker compound is provided, comprising TIFF2026503087000015.tif22139 or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; Each R 3 are independently bonded, C1 to C 12Alkylene, -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)-, -C1-C 12 Alkylene-NR a -C(O)-, -C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -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)-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-, -C(O)-NR a -C1~C 12 Alkylene-(CH(OH)) 1~8 -C1~C 12 Alkylene-, -O-CH2-CH2, -OC(O)-NR a -C1~C 12 Alkylene, -O-CH2-CH(OH)-C(O)-, -O-CH2-CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C1-C 12 Alkylene, C1-C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C1~C 12Alkylene-, -CH(OH)-C1-C 12 Alkylene-NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-NR a -C1~C 12 Alkylene-, -[C(O)-(CH2) 1~8 -NR a ] 1~8 -, Triazolyl, -C1~C 12 Alkylene-triazolyl-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, and each R a are independently H, C 1~6 alkyl, wherein any of the above alkylene groups may be substituted with —SO3H; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R 6 are independently a bond or: (i) TIFF2026503087000016.tif26128In the above formula, each n 3 and n4 are independently between 0 and 1, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; m is 1 to 4; each v is independently 1 to 6; n 2 is 1; (ii) TIFF2026503087000017.tif19128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, n 6 is 1 to 10, each p is independently 0 to 6; n 2 is 1; (iii) TIFF2026503087000018.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; q is 1 to 8; each v is independently 1 to 6; n 2 is 1; (iv) TIFF2026503087000019.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, each p is independently 0 to 6; n 2 is 1; (v) -R 10 -[O-CH2-CH2] 1~8 -R 10 - In the above formula, Each R b are independently H or C 1~6 is alkyl, Each R 10 is independent TIFF2026503087000020.tif24128, each p is independently 1 to 6; q is 1 to 8; and (vi) -N-(R 1 -XR 2 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 )2 In the above formula, Each X is independently -NR a -C(O)- or -C(O)NR a - and n 2 is 2; More selected; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; each n 0 are independently 2 to 26; n 1 is 0 to 6, and n 1 If is 0, R 3 is -OH or -C(O)ORb where R b are independently H or C 1~6 is alkyl; n 3 is 1 to 6.

[0013] In some embodiments, a Drug-Linker Compound is provided that comprises a linker compound described herein having at least one Drug unit attached thereto.

[0014] In some embodiments, a conjugate is provided that includes a targeting unit attached to a drug-linker compound described herein.

[0015] In some aspects, a pharmaceutical composition is provided comprising a conjugate described herein and a pharmaceutically acceptable carrier.

[0016] 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 molecule, such as a target antigen, associated with the cancer or autoimmune disease.

[0017] 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]

[0018] The present disclosure will be further described based on exemplary embodiments. These exemplary embodiments will be described in detail with reference to the drawings. It should be noted that the drawings are not to scale. These embodiments are non-limiting exemplary embodiments, and like reference numerals represent like structures throughout the multiple views of the drawings.

Figure 1

Figure 2

Figure 3

Figure 4

[0019] 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.

[0020] 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.

[0021] 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.

[0022] 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.

[0023] 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.

[0024] 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.

[0025] 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.

[0026] 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.

[0027] 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)).

[0028] 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).

[0029] 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.

[0030] 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.

[0031] 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-).

[0032] 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.

[0033] 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, saturated and containing 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 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.

[0034] 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.

[0035] 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, no orientation is implied for alkylene and heteroalkylene.

[0036] Unless otherwise stated, the terms "heteroalkenylene" and "heteroalkynylene," by themselves or as part of other terms, 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, no orientation is implied for heteroalkenylene and heteroalkynylene.

[0037] 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.

[0038] 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).

[0039] 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).

[0040] 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.

[0041] 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).

[0042] 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.

[0043] 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.

[0044] 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.

[0045] 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).

[0046] Unless otherwise indicated, "carboxyl" is COOH or COO - M + where M + is a cation.

[0047] Unless otherwise indicated, "oxo" refers to (C=O).

[0048] Unless otherwise indicated, "substituted alkyl" and "substituted aryl" refer to alkyl and aryl, respectively, in which one or more hydrogen atoms are each 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.

[0049] 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.

[0050] 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.

[0051] Any substituents replacing a hydrogen in any one 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( =O)2N(R')R", -S(=O)2NH2, -S(=O)2N(R')R", -S(=O)2OR', -S(=O)R", -OP(=O)(OR')(OR" ), -OP(OH)3, -P(=O)(OR')(OR"), -PO3H2, -C(=O)R', -C(=S)R", -CO2R', -C(=S)OR", -C(=O )SR', -C(=S)SR', -C(=S)NH2, -C(=S)N(R')(R")2, -C(=NR')NH2, -C(=NR')N(R')R" and salts thereof, wherein each X is independently selected from the group consisting of halogen: -F, -CI, -Br, and -I; and wherein each R" is independently C1 to C 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.

[0052] 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-C21 alkyl, C19-C22 alkyl, C19-C25 alkyl, C19-C26 alkyl, C19-C28 alkyl, C19-C29 alkyl, C19-C25 alkyl, C19-C26 alkyl, C19-C27 alkyl, C19-C28 alkyl, C19-C29 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.

[0053] 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.

[0054] "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.

[0055] 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.

[0056] 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 a pharmaceutically acceptable salt contains multiple charged atoms, multiple counterions may be present. Thus, a pharmaceutically acceptable salt may have one or more charged atoms and / or one or more counterions.

[0057] 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.

[0058] 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.

[0059] 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%.

[0060] 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.

[0061] Other terms are defined herein within the description of various aspects of the invention.

[0062] Detailed Description Provided herein are linkers comprising polar groups such as saccharide units, polymer units, and / or carboxyl units. Also provided are targeting unit-linkers, drug linkers, and conjugates thereof, comprising drug units such as cytotoxic agents or immunomodulatory agents, as further described herein.

[0063] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 6-([O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 ) n3 (Ia) or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; Each R 3 are independently bonded, C1 to 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)-, -C1-C 12 Alkylene-NR a -C(O)-, -C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -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)-, -NR a -C(O)-C1~C 12Alkylene-C(O)-, -C(O)-NR a -C1~C 12 Alkylene-(CH(OH)) 1~8 -C1~C 12 Alkylene-, -O-CH2-CH2, -OC(O)-NR a -C1~C 12 Alkylene, -O-CH2-CH(OH)-C(O)-, -O-CH2-CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C1-C 12 Alkylene, C1-C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-C1-C 12 Alkylene-NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-NR a -C1~C 12 Alkylene-, -[C(O)-(CH2) 1~8 -NR a ] 1~8 -, Triazolyl, -C1~C 12 Alkylene-triazolyl-, -N(polyhydroxyl group)-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, and each R a are independently H, C 1~6 alkyl, wherein any of the above alkylene groups may be substituted with —SO3H; Each R 4and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R 6 teeth, (i) TIFF2026503087000021.tif26128In the above formula, each n 3 and n 4 are independently between 0 and 1, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; m is 1 to 4; each v is independently 1 to 6; n 2 is 1; (ii) TIFF2026503087000022.tif20128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, n 6 is 1 to 10, each p is independently 0 to 6; n 2 is 1; (iii) TIFF2026503087000023.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; q is 1 to 8; each v is independently 1 to 6; n 2 is 1; (iv) TIFF2026503087000024.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, each p is independently 0 to 6; n 2 is 1; (v) -R 10 -[O-CH2-CH2] 1~8 -R 10 - In the above formula, Each R b are independently H or C 1~6 is alkyl, Each R 10 is independent TIFF2026503087000025.tif24128, each p is independently 1 to 6; Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), q is 1 to 8; n 2 is 1; (vi) -N-(R1 -XR 2 -)2 In the above formula, Each X is independently -NR a -C(O)- or -C(O)NR a - and n 2 is 2; More selected; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; each n 0 are independently 2 to 26; each n 1 are independently 1 to 6; n 3 is 1 to 6, polar group A linker compound is provided, comprising:

[0064] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 6 -([O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 ) n3 (Ia) or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2are independently a bond or C1-C6 alkylene; Each R 3 are independently bonded, C1 to 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)-, -C1-C 12 Alkylene-NR a -C(O)-, -C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -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)-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-, -C(O)-NR a -C1~C 12 Alkylene-(CH(OH)) 1~8 -C1~C 12 Alkylene-, -O-CH2-CH2, -OC(O)-NR a -C1~C 12 Alkylene, -O-CH2-CH(OH)-C(O)-, -O-CH2-CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C1-C 12 Alkylene, C1-C12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-C1-C 12 Alkylene-NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-NR a -C1~C 12 Alkylene-, -[C(O)-(CH2) 1~8 -NR a ] 1~8 -, Triazolyl, -C1~C 12 Alkylene-triazolyl-, -N(polyhydroxyl group)-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, and each R a are independently H, C 1~6 alkyl, wherein any of the above alkylene groups may be substituted with —SO3H; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether 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, -C(O)-R where R is a saccharide unit of formula (XII) or (XIII), a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R 6 teeth, (i) TIFF2026503087000026.tif26128In the above formula, each n 3 and n 4 are independently between 0 and 1, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; m is 1 to 4; each v is independently 1 to 6; n 2 is 1; (ii) TIFF2026503087000027.tif20128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, n 6 is 1 to 10, each p is independently 0 to 6; n 2 is 1; (iii) TIFF2026503087000028.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; q is 1 to 8; each v is independently 1 to 6; n 2 is 1; (iv) TIFF2026503087000029.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, each p is independently 0 to 6; n 2 is 1; (v) -R 10 -[O-CH2-CH2] 1~8 -R 10 - In the above formula, Each R b are independently H or C 1~6 is alkyl, Each R 10 is independent TIFF2026503087000030.tif24128, each p is independently 1 to 6; Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), q is 1 to 8; n 2is 1; and (vi) -N-(R 1 -XR 2 -)2 In the above formula, Each X is independently -NR a -C(O)- or -C(O)NR a - and n 2 is 2; More selected; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; each n 0 are independently 2 to 26; each n 1 are independently 1 to 6; n 3 is 1 to 6, polar group A linker compound is provided, comprising:

[0065] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 2 -(NR 4 R 5 ) n1 ) n3 (Ia') or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; Each R 3 are independently -N(polyhydroxyl group), triazolyl, -C1 to -C 12 alkylene-triazolyl-, TIFF2026503087000031.tif101128; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R a are independently H or C 1~6 is alkyl; TIFF2026503087000032.tif8128 is R 0 to R 3 indicates the binding site of; Wavy line TIFF2026503087000033.tif8128 is R 1 to R 3 indicates the binding site of; each p is 1 to 6; each n 0 are independently 2 to 8; each n 1 are independently 1 to 6; n 3 is 1 to 6, polar group A linker compound is provided, comprising:

[0066] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 2 -(NR 4 R 5 ) n1 ) n3 (Ia') or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; Each R 3 are independently -N(polyhydroxyl group), triazolyl, -C1 to -C 12 alkylene-triazolyl-, TIFF2026503087000034.tif101128; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether 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, -C(O)-R where R is a saccharide unit of formula (XII) or (XIII), a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R5 At least one of them is not H; Each R a are independently H or C 1~6 is alkyl; TIFF2026503087000035.tif8128 is R 0 to R 3 indicates the binding site of; Wavy line TIFF2026503087000036.tif8128 is R 1 to R 3 indicates the binding site of; each p is 1 to 6; each n 0 are independently 2 to 8; each n 1 are independently 1 to 6; n 3 is 1 to 6, polar group A linker compound is provided, comprising:

[0067] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 1 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n3 (Ia") or a stereoisomer or salt thereof, wherein: (i) R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2are independently a bond or C1-C6 alkylene; R 3 is -C(O)-; R 4 is H; R 5 is independently a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; n 0 are independently 2 to 26; n 1 is 1 to 6; n 3 is 1-6; (ii) R 0 is -C(O)-; R 1 , R 2 , and R 3 are bonds respectively; R 4 and R 5 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; n 0 is 6; n 1 is 1 to 6; n 3 is 1; (iii) R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C6 alkylene; R 3 Ha-NR a -C(O)-C1~C 12 alkylene-C(O)-, where the alkylene is substituted with -SO3H; R a is H or C 1~6 is alkyl; R 4 and R 5 are each independently H, a carboxyl-containing moiety, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; each n 0 are independently 1 to 26; n 1 is 1 to 6; n 3 is 1 to 6; or (iv) R 0 teeth TIFF2026503087000037.tif24128; Each R 1 are independently a bond or C1-C6 alkylene; R 2 and R 3 are bonds respectively; R 4 and R 5are each independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R a are independently H or C 1~6 is alkyl; Wavy line TIFF2026503087000038.tif7128 is the R to the rest of the polymer units 0 indicates the binding site of; Wavy line (~*) is R 0 indicates the binding site of the amino acid unit to; n 0 is 1 to 8; n 1 is 1 to 6; n 3 is 2, polar group A linker compound is provided, comprising:

[0068] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 1 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n3 (Ia") or a stereoisomer or salt thereof, wherein: (i) R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; R 3 is -C(O)-; R 4 is H; R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether 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, -C(O)-R where R is a saccharide unit of formula (XII) or (XIII), a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; n 0 are independently 2 to 26; n 1 is 1 to 6; n 3 is 1-6; (ii) R 0 is -C(O)-; R 1 , R 2 , and R 3 are bonds respectively; R 4 and R 5are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl ether group, a substituted polyhydroxyl ether group, or an optionally substituted C3 to 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, -C(O)-R where R is a saccharide unit of formula (XII) or (XIII), a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; n 0 is 6; n 1 is 1 to 6; n 3 is 1; (iii) R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C6 alkylene; R 3 Ha-NR a -C(O)-C1~C 12 alkylene-C(O)-, where the alkylene is substituted with -SO3H; R a is H or C 1~6 is alkyl; R 4 and R 5are each independently H, a carboxyl-containing moiety, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether 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, -C(O)-R where R is a saccharide unit of formula (XII) or (XIII), a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; each n 0 are independently 1 to 26; n 1 is 1 to 6; n 3 is 1 to 6; or (iv) R 0 teeth TIFF2026503087000039.tif24128; Each R 1 are independently a bond or C1-C6 alkylene; R 2 and R 3 are bonds respectively; R 4 and R 5 are each independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, a chelating agent, or —NR 4 R5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R a are independently H or C 1~6 is alkyl; Wavy line TIFF2026503087000040.tif7128 is the R to the rest of the polymer units 0 indicates the binding site of; Wavy line (~*) is R 0 indicates the binding site of the amino acid unit to; n 0 is 1 to 8; n 1 is 1 to 6; n 3 is 2, polar group A linker compound is provided, comprising:

[0069] In some embodiments, the following: (a) A linker unit having 1 to 4 attachment sites per drug unit, wherein the linker unit has the formula: TIFF2026503087000041.tif49128 or a stereoisomer or salt thereof, wherein: α- represents the direct or indirect attachment site to the amino acid unit; δ- represents an attachment site for at least one of the Drug units or for a linking group attached to at least one of the Drug units; R a is H or C 1~6 is alkyl, Linker units; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to an amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit; A linker compound is provided, comprising:

[0070] In some embodiments, the following: (a) A linker unit having 1 to 4 attachment sites per drug unit, wherein the linker unit has the formula: TIFF2026503087000042.tif49128 or a stereoisomer or salt thereof, wherein: α- represents the direct or indirect binding site to an amino acid; δ- represents an attachment site to at least one of the Drug units or for a linking group attached to at least one of the Drug units; R a is H or C 1~6 is alkyl, Linker units; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to an amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit; A linker compound is provided, comprising:

[0071] In some embodiments, the linker unit has the formula: TIFF2026503087000043.tif49128 or a stereoisomer or salt thereof.

[0072] In some embodiments, the linker unit has the formula: TIFF2026503087000044.tif47128 or a stereoisomer or salt thereof.

[0073] In some embodiments, the linker unit has the formula: TIFF2026503087000045.tif50128 or a stereoisomer or salt thereof.

[0074] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit comprising: (i) optionally substituted polyamides; (ii) a substituted polyether; or (iii) any combination thereof Including, polar group A linker compound is provided, comprising:

[0075] In some embodiments, the following: (a) a linker unit with 1 to 4 binding sites per drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit comprising: (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, Polyamides containing TIFF2026503087000046.tif20128, or stereoisomers thereof; (ii) Each R b are independently H or C 1~6 is alkyl, and n 0 are independently 2 to 26, a polyether containing TIFF2026503087000047.tif22128, or a stereoisomer thereof; or (iii) any combination thereof Including, polar group A linker compound is provided, comprising:

[0076] In some embodiments, at least one polar group attached to an amino acid unit has the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 6 -([O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 ) n3 (Ia), ~R 0 -(R 3 -R 1 -[O-CH2-CH(OH)-CH2] n0 -R 6 -[O-CH2-CH(OH)-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n3 (Ib), or A linker compound is provided, comprising TIFF2026503087000048.tif22138 or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; Each R 3 are independently bonded, C1 to 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)-, -C1-C 12 Alkylene-NR a-C(O)-, -C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -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)-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-, -C(O)-NR a -C1~C 12 Alkylene-(CH(OH)) 1~8 -C1~C 12 Alkylene-, -O-CH2-CH2, -OC(O)-NR a -C1~C 12 Alkylene, -O-CH2-CH(OH)-C(O)-, -O-CH2-CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C1-C 12 Alkylene, C1-C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-C1-C 12 Alkylene-NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a-C1~C 12 Alkylene-, -CH(OH)-NR a -C1~C 12 Alkylene-, -[C(O)-(CH2) 1~8 -NR a ] 1~8 -, Triazolyl, -C1~C 12 Alkylene-triazolyl-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, and each R a are independently H, C 1~6 alkyl, wherein any of the above alkylene groups may be substituted with —SO3H; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R 6 are independently a bond or: (i) TIFF2026503087000049.tif27128In the above formula, each n 3 and n 4 are independently between 0 and 1, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; m is 1 to 4; each v is independently 1 to 6; n 2 is 1; (ii) TIFF2026503087000050.tif20128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, n 6 is 1 to 10, each p is independently 0 to 6; n 2 is 1; (iii) TIFF2026503087000051.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; q is 1 to 8; each v is independently 1 to 6; n 2 is 1; (iv) TIFF2026503087000052.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, each p is independently 0 to 6; n 2is 1; (v) -R 10 -[O-CH2-CH2] 1~8 -R 10 - In the above formula, Each R b are independently H or C 1~6 is alkyl, Each R 10 is independent TIFF2026503087000053.tif24128, each p is independently 1 to 6; q is 1 to 8; and (vi) -N-(R 1 -XR 2 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 )2 In the above formula, Each X is independently -NR a -C(O)- or -C(O)NR a - and n 2 is 2; More selected; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; each n 0 are independently 2 to 26; n 1 is 0 to 6, and n 1 If is 0, R 3 is -OH or -C(O)OR b where R b are independently H or C 1~6 is alkyl; n 3 is 1 to 6.

[0077] In some embodiments, at least one polar group attached to an amino acid unit has the formula: ~R 0 -(R 3-R 1 -[O-CH2-CH2] n0 -R 6 -([O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 ) n3 (Ia), ~R 0 -(R 3 -R 1 -[O-CH2-CH(OH)-CH2] n0 -R 6 -[O-CH2-CH(OH)-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n3 (Ib), or A linker compound is provided, comprising TIFF2026503087000054.tif22139 or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C1-C6 alkylene; Each R 3 are independently bonded, C1 to 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)-, -C1-C 12 Alkylene-NR a -C(O)-, -C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -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)-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-, -C(O)-NR a -C1~C 12 Alkylene-(CH(OH)) 1~8 -C1~C 12 Alkylene-, -O-CH2-CH2, -OC(O)-NR a -C1~C 12 Alkylene, -O-CH2-CH(OH)-C(O)-, -O-CH2-CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C1-C 12 Alkylene, C1-C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-C1-C 12 Alkylene-NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-NR a -C1~C 12 Alkylene-, -[C(O)-(CH2)1~8 -NR a ] 1~8 -, Triazolyl, -C1~C 12 Alkylene-triazolyl-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, and each R a are independently H, C 1~6 alkyl, wherein any of the above alkylene groups may be substituted with —SO3H; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether 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, -C(O)-R where R is a saccharide unit of formula (XII) or (XIII), a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R 6 are independently a bond or: (i) TIFF2026503087000055.tif27128In the above formula, each n 3 and n 4 are independently between 0 and 1, Each R b are independently H or C1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; m is 1 to 4; each v is independently 1 to 6; n 2 is 1; (ii) TIFF2026503087000056.tif20128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, n 6 is 1 to 10, each p is independently 0 to 6; n 2 is 1; (iii) TIFF2026503087000057.tif24128In the above formula, Each R a are independently H or C 1~6 is alkyl, Each R b are independently H or C 1~6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH)2, or -(CH2) v -OS(=O)2(OH), each p is independently 0 to 6; q is 1 to 8; each v is independently 1 to 6; n 2 is 1; (iv) TIFF2026503087000058.tif24128In the above formula, Each R a are independently H or C 1~6is alkyl, Each R b are independently H or C 1~6 is alkyl, each p is independently 0 to 6; n 2 is 1; (v) -R 10 -[O-CH2-CH2] 1~8 -R 10 - In the above formula, Each R b are independently H or C 1~6 is alkyl, Each R 10 is independent TIFF2026503087000059.tif24128, each p is independently 1 to 6; q is 1 to 8; and (vi) -N-(R 1 -XR 2 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 )2 In the above formula, Each X is independently -NR a -C(O)- or -C(O)NR a - and n 2 is 2; More selected; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to; each n 0 are independently 2 to 26; n 1 is 0 to 6, and n 1 If is 0, R 3 is -OH or -C(O)OR b where R b are independently H or C 1~6 is alkyl; n 3is 1 to 6.

[0078] In some embodiments, each R 3 are independently a bond, -C(O)-, or -NR a -C(O)-C1~C 12 Alkylene-C(O)-, -C(O)-NR a -C1~C 12 Alkylene-(CH(OH)) 1~8 -C1~C 12 Alkylene-, -O-CH2-CH(OH)-C(O)-, -O-CH2-CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C1-C 12 Alkylene, C1-C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-C1-C 12 Alkylene-NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-NR a -C1~C 12 Alkylene-, -[C(O)-(CH2) 1~8 -NR a ] 1~8 -, triazolyl, and -C1 to C 12 alkylene-triazolyl-, -N(polyhydroxyl group)-, and each R a are independently H, C 1~6 Linker compounds are provided in which any of the above alkylene groups may be substituted with -SO3H.

[0079] In some embodiments, each R 3are independently a bond, -C(O)-, or -NR a -C(O)-C1~C 12 Alkylene-C(O)-, -C(O)-NR a -C1~C 12 Alkylene-(CH(OH)) 1~8 -C1~C 12 Alkylene-, -O-CH2-CH(OH)-C(O)-, -O-CH2-CH(OH)-C(O)-NR a -C1~C 12 Alkylene, C1-C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C1~C 12 Alkylene-, -CH(OH)-C1-C 12 Alkylene-NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -NR a -C(O)-C1~C 12 Alkylene-C(O)-NR a -C1~C 12 Alkylene-, -[C(O)-(CH2) 1~8 -NR a ] 1~8 -, triazolyl, and -C1 to C 12 alkylene-triazolyl-, -N(polyhydroxyl group)-, and each R a are independently H, C 1~6 Linker compounds are provided in which any of the above alkylene groups may be substituted with -SO3H.

[0080] In some aspects, the linker unit is A linker compound is provided, comprising a moiety selected from TIFF2026503087000060.tif58134 or a stereoisomer or salt thereof; During the ceremony, α- represents the direct or indirect attachment site to the amino acid unit; d- represents a site of attachment to at least one of the Drug units or to a linking group attached to at least one of the Drug units; R a is H or C 1~6 It is alkyl.

[0081] In some embodiments, at least one polar group has the formula: L3-N(CH2-(CH(X1R)) k -X2(X3))2(X) or a stereoisomer or salt thereof, During the ceremony, each X1 is independently selected from NH or O; each R is independently selected from hydrogen, acetyl, monosaccharides, disaccharides, and polysaccharides; each X2 is independently selected from CH2 and C(O); each X3 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.

[0082] In some embodiments, at least one polar group has the following structure (XII) or (XIII): TIFF2026503087000061.tif84128 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.

[0083] In some embodiments, the following: (a) ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -NR 4 R 5 (XX) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C3 alkylene; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5 Both of them cannot be H; n0 is 2 to 26; (b) ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -NR 4 R 5 (XXI) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C3 alkylene; R 4 and R 5 is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), and R 4 and R 5 The other is a polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, wherein R 4 and R 5 Both of them cannot be H; n0 is 2 to 26; (c) ~R 0 -[-R 6 -[R 9 -[O-CH2-CH2-] n0 -R 9 ] n1 -R 7 -NR 4 R 5 ] n7 (XXII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 6 and R 7 are each independently a bond, 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-C 12 alkylene-NH-; R 4 and R 5 is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a chelating agent, and —C(O)—R, where R is a saccharide unit of formula (XII) or (XIII); and R 4 and R 5 the other is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), and polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, wherein R 4 and R 5 Both of them cannot be H; Each R 9is independently selected from a bond, —C(O)—, —NH—, —C(O)—C-C alkylene-, —NH—C-C alkylene-, —C-C alkylene-NH—, —C-C alkylene-C(O)—, —NH(CO)—C-C alkylene-, —N(CH)—(CO)—C-C alkylene-, —NH(CO)NH—, and triazole; n0 is 2 to 26; n1 is 1 to 4; n7 is 1-4; (d) ~R 0 -R 1 -[-C(R α )HC(O)-N(R N )-] n0 -R 2 -NR 4 R 5 (XXIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 is a bond, C1-C3 alkylene, -C1-C3 alkylene-[O-CH2-CH2-] n0 , -[CH2-CH2-O] n0 -C1-C3 alkylene-, or -C1-C3 alkylene-[O-CH2-CH2-] n0 -C(O)-; R 2 is C1-C3 alkylene, -C1-C3 alkylene-[O-CH2-CH2-] n0 , -[CH2-CH2-O] n0 -C1-C3 alkylene-, or -C1-C3 alkylene-[O-CH2-CH2-] n0 -C(O)-; Each R α are independently H or -R 2 -NR 4 R 5 and; Each R N are independently H, C1-C6 alkyl, or -R 2 -NR 4 R 5and; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5 Both of them cannot be H; Each n0 is independently 2 to 26; (e) ~R 0 -R 1 -[-C(R α )HC(O)-N(R N )-] n0 -R 2 -CO2R 6 (XXIV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 is a bond, C1-C3 alkylene, or -C1-C3 alkylene [O-CH2-CH2-] n0 and; R 2 is C1-C3 alkylene or -C1-C3 alkylene [O-CH2-CH2-] n0 and; Each R α are independently H or -R 2 -NR 4 R 5 and; Each R N are independently H, C1-C6 alkyl, or -R 2 -NR 4 R 5 and; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII),4 and R 5 Both of them cannot be H; R 6 is H or C1-C4 alkyl; Each n0 is independently 2 to 26, However, at least one R α or R N Ha-R 2 -NR 4 R 5 is; or (f) ~R 0 -R 1 -[C(R α )HC(O)-N(R N )-] n0 -R 2 -N-(R 3 -NR 4 R 5 )2(XXV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond, C1-C3 alkylene, or -C1-C3 alkylene-[O-CH2-CH2-] n0 and; Each R α are independently H or -R 2 -NR 4 R 5 and; Each R N are independently H or C1-C6 alkyl; Each R 3 are independently C1-C6 alkylene; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5Both of them cannot be H; Each n0 is independently 2 to 26; Linker compounds are provided that include a polar group having a formula selected from:

[0084] In some embodiments, the following: (a) ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -NR 4 R 5 (XX) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C3 alkylene; R 4 and R 5 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl ether group, a substituted polyhydroxyl ether group, or an optionally substituted C3 to 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, -C(O)-R, where R is a sugar unit of formula (XII) or (XIII), and a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; n0 is 2 to 26; (b) ~R 0 -R 1-[O-CH2-CH2] n0 -R 2 -NR 4 R 5 (XXI) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C3 alkylene; R 4 and R 5 is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), and R 4 and R 5 The other is a polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, wherein R 4 and R 5 Both of them cannot be H; n0 is 2 to 26; (c) ~R 0 -[-R 6 -[R 9 -[O-CH2-CH2-] n0 -R 9 ] n1 -R 7 -NR 4 R 5 ] n7 (XXII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 6 and R 7 are each independently a bond, C1 to C 12 Alkylene, -NH-C1~C 12 Alkylene, -C1~C 12 Alkylene-NH-, -C1~C 12Alkylene -N(CH3)-, -C(O)-C1~C 12 Alkylene, -C1~C 12 Alkylene-C(O)-, -NH-C1-C 12 Alkylene -C(O)- and -C(O)-C1-C 12 alkylene-NH-; R 4 and R 5 is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII); and R 4 and R 5 the other is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), and polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, or -NR 4 R 5 together form a C3-C8 heterocycle, and R 4 and R 5 Both of them cannot be H; Each R 9 is independently selected from a bond, —C(O)—, —NH—, —C(O)—C-C alkylene-, —NH—C-C alkylene-, —C-C alkylene-NH—, —C-C alkylene-C(O)—, —NH(CO)—C-C alkylene-, —N(CH)—(CO)—C-C alkylene-, —NH(CO)NH—, and triazole; n0 is 2 to 26; n1 is 1 to 4; n7 is 1-4; (d) ~R 0 -R 1 -[-C(R α )HC(O)-N(R N )-] n0 -R 2 -NR 4 R5 (XXIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 is a bond, C1-C3 alkylene, -C1-C3 alkylene-[O-CH2-CH2-] n0 , -[CH2-CH2-O] n0 -C1-C3 alkylene-, or -C1-C3 alkylene-[O-CH2-CH2-] n0 -C(O)-; R 2 is C1-C3 alkylene, -C1-C3 alkylene-[O-CH2-CH2-] n0 , -[CH2-CH2-O] n0 -C1-C3 alkylene-, or -C1-C3 alkylene-[O-CH2-CH2-] n0 -C(O)-; Each R α are independently H or -R 2 -NR 4 R 5 and; Each R N are independently H, C1-C6 alkyl, or -R 2 -NR 4 R 5 and; R 4 and R 5 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl ether group, a substituted polyhydroxyl ether group, or an optionally substituted C3 to 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, -C(O)-R, where R is a sugar unit of formula (XII) or (XIII), and a chelating agent, or -NR 4 R 5together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each n0 is independently 2 to 26; (e) ~R 0 -R 1 -[-C(R α )HC(O)-N(R N )-] n0 -R 2 -CO2R 6 (XXIV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 is a bond, C1-C3 alkylene, or -C1-C3 alkylene [O-CH2-CH2-] n0 and; R 2 is C1-C3 alkylene or -C1-C3 alkylene [O-CH2-CH2-] n0 and; Each R α are independently H or -R 2 -NR 4 R 5 and; Each R N are independently H, C1-C6 alkyl, or -R 2 -NR 4 R 5 and; R 4 and R 5 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl ether group, a substituted polyhydroxyl ether group, or an optionally substituted C3 to 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, -C(O)-R, where R is a sugar unit of formula (XII) or (XIII), and a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; R 6 is H or C1-C4 alkyl; Each n0 is independently 2 to 26, However, at least one R α or R N Ha-R 2 -NR 4 R 5 is; or (f) ~R 0 -R 1 -[C(R α )HC(O)-N(R N )-] n0 -R 2 -N-(R 3 -NR 4 R 5 )2(XXV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond, C1-C3 alkylene, or -C1-C3 alkylene-[O-CH2-CH2-] n0 and; Each R α are independently H or -R 2 -NR 4 R 5 and; Each R N are independently H or C1-C6 alkyl; Each R 3are independently C1-C6 alkylene; R 4 and R 5 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a polyhydroxyl ether group, a substituted polyhydroxyl ether group, or an optionally substituted C3 to 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, -C(O)-R, where R is a sugar unit of formula (XII) or (XIII), and a chelating agent, or -NR 4 R 5 together form a C3-C8 heterocycle, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each n0 is independently 2 to 26; Linker compounds are provided that include a polar group having a formula selected from:

[0085] In some embodiments, R 4 and R 5 are each independently selected from H and a polyhydroxyl group, where R 4 and R 5 is not H.

[0086] In some embodiments, linker compounds are provided in which the polyhydroxyl group is a linear monosaccharide optionally selected from a C6 or C5 sugar, a sugar acid, or an amino sugar.

[0087] In some embodiments, 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; A linker compound is provided.

[0088] In some embodiments, the following: TIFF2026503087000062.tif230121TIFF2026503087000063.tif193144TIFF2026503087000 064.tif248148TIFF2026503087000065.tif250158TIFF2026503087000066.tif223158TIFF2 026503087000067.tif243159TIFF2026503087000068.tif212158TIFF2026503087000069.tif201159TIFF2026503087000070.tif63128 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; and the wavy line is a bond to an amino acid unit.

[0089] In some embodiments, R 4 and R 5 is provided, wherein one of the two is a linear monosaccharide and the other is a cyclic monosaccharide.

[0090] In some embodiments, —(NR4 R 5 ) but the following: TIFF2026503087000071.tif35128 or a stereoisomer or salt thereof; In the formula, R 11 is a cyclic monosaccharide.

[0091] In some embodiments, the following: a linker compound comprising a polar group selected from TIFF2026503087000072.tif32128TIFF2026503087000073.tif70128 or a stereoisomer or salt thereof; In the formula, R 41 is a cyclic monosaccharide; the wavy lines indicate the bonds to the amino acid units.

[0092] In some embodiments, R 4 and R 5 are independently a polyhydroxyl selected from cyclic monosaccharides, disaccharides, and polysaccharides.

[0093] In some embodiments, —(NR 4 R 5 ) but the following: TIFF2026503087000074.tif104133 or a stereoisomer or salt thereof; In the formula, each R 12 is selected from H and a monosaccharide, a disaccharide, or a polysaccharide; R 5 is selected from cyclic monosaccharides, disaccharides, or polysaccharides.

[0094] In some embodiments, the following: TIFF2026503087000075.tif190144 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 46is selected from cyclic monosaccharides, disaccharides, or polysaccharides; the wavy line indicates the bond to an amino acid unit.

[0095] In some embodiments, R 4 and R 5 are independently selected from linear monosaccharides and substituted linear monosaccharides, wherein the substituted linear monosaccharide is replaced with a monosaccharide, a disaccharide, or a polysaccharide.

[0096] In some embodiments, —(NR 4 R 5 ) but the following: TIFF2026503087000076.tif38128 or a stereoisomer or salt thereof; In the formula, R 13 is a linear monosaccharide; each R 14 is selected from monosaccharides, disaccharides and polysaccharides.

[0097] In some embodiments, the following: a linker compound comprising a polar group selected from TIFF2026503087000077.tif82128TIFF2026503087000078.tif67128 or a stereoisomer or salt thereof; In the formula, R 47 is a linear monosaccharide; each R 49 are selected from monosaccharides, disaccharides and polysaccharides; the wavy lines are bonds to amino acid units.

[0098] In some embodiments, R 4 and R 5 are 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 with a monosaccharide, disaccharide, or polysaccharide.

[0099] In some embodiments, R 4 and R5 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 be further substituted with a monosaccharide, disaccharide, or polysaccharide.

[0100] In some embodiments, —(NR 4 R 5 ) but the following: TIFF2026503087000079.tif38128 or a stereoisomer or salt thereof; In the formula, each R 15 are independently selected from linear monosaccharides and substituted linear monosaccharides; each R 16 is independently selected from carboxyl, ester, and amide.

[0101] In some embodiments, —(NR 4 R 5 ) but the following: TIFF2026503087000080.tif38128 or a stereoisomer or salt thereof; In the formula, each R 15 are independently selected from linear monosaccharides and substituted linear monosaccharides; each R 16 is independently selected from alkyl, O-alkyl, aryl, O-aryl, carboxyl, ester, and amide.

[0102] In some embodiments, the following: TIFF2026503087000081.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 43are independently selected from carboxyl, ester, and amide; the wavy line is the bond to the amino acid unit.

[0103] In some embodiments, the following: TIFF2026503087000082.tif85128 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 is the bond to the amino acid unit.

[0104] In some embodiments, R 4 and R 5 is a -C(O)-polyhydroxyl group or a substituted -C(O)-polyhydroxyl group, and R 4 and R 5 and 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 the polyhydroxyl group are substituted with a monosaccharide, disaccharide, polysaccharide, carboxyl, ester, or amide.

[0105] In some embodiments, R 4 and R 5 is a -C(O)-polyhydroxyl group or a substituted -C(O)-polyhydroxyl group, and R 4 and R 5 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, disaccharide, polysaccharide, alkyl, -O-alkyl, aryl, carboxyl, ester, or amide.

[0106] In some embodiments, —(NR 4 R 5 ) but the following: TIFF2026503087000083.tif39128 or a stereoisomer or salt thereof.

[0107] In some embodiments, the following: TIFF2026503087000084.tif81128 or a stereoisomer or salt thereof, where the wavy lines are bonds to the amino acid units.

[0108] In some embodiments, R 4 and R 5 are independently selected from H, substituted -C1-C8 alkyl, substituted -C1-C4 alkyl, or substituted -C1-C3 alkyl; 4 and R 5 at least one of is not H; wherein the substituted -C1-C8 alkyl, -C1-C4 alkyl and -C1-C3 alkyl are substituted with hydroxyl and / or carboxyl.

[0109] In some embodiments, —(NR 4 R 5 ) but the following: TIFF2026503087000085.tif54151 or a stereoisomer or salt thereof; In the formula, R 18 is selected from OH, CH2OH, COOH, or -C1-C6 alkyl substituted with hydroxyl or carboxyl.

[0110] In some embodiments, —(NR 4 R 5 ) but the following: TIFF2026503087000086.tif54151 or a stereoisomer or salt thereof; In the formula, R 18 is selected from H, OH, CH2OH, COOH, or -C1-C6 alkyl substituted with hydroxyl or carboxyl.

[0111] In some embodiments, the following: a linker compound comprising a polar group selected from TIFF2026503087000087.tif129128TIFF2026503087000088.tif98128 or a stereoisomer or salt thereof; In the formula, R 48 is selected from OH, CH2OH, COOH, or -C1-C6 alkyl substituted with hydroxyl or carboxyl; the wavy line is the bond to the amino acid unit.

[0112] In some embodiments, the following: a linker compound comprising a polar group selected from TIFF2026503087000089.tif111128TIFF2026503087000090.tif123128 or a stereoisomer or salt thereof; In the formula, R 48 is selected from H, OH, CH2OH, COOH, or -C1-C6 alkyl substituted with hydroxyl or carboxyl; the wavy line is the bond to the amino acid unit.

[0113] In some embodiments, R 4 and R 5 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 4 and R 5is 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.

[0114] In some embodiments, —(NR 4 R 5 ) but the following: TIFF2026503087000091.tif35158TIFF2026503087000092.tif21146 or a stereoisomer or salt thereof.

[0115] In some embodiments, the following: a linker compound comprising a polar group selected from TIFF2026503087000093.tif179128TIFF2026503087000094.tif78128 or a stereoisomer or salt thereof; where the wavy lines are bonds to the amino acid units.

[0116] In some embodiments, the R of the polymer unit 24 and R 25 is selected from H and optionally substituted aryl; provided that R 24 and R 25 and n are not both H, where the optional substituents are as defined herein, e.g., in some embodiments, the optional substituents are halo, such as F, Cl, or Br. In some embodiments, a linker intermediate or linker is provided in which the polymer unit is selected from the group consisting of: a linker intermediate or linker selected from TIFF2026503087000095.tif71128 or a salt thereof; In the formula, the wavy line on the left indicates the site of attachment of the amino acid unit to a subunit or part of the linker subunit.

[0117] In some embodiments, R 4 and R 5 taken together form an optionally substituted C3-C8 heterocycle or heteroaryl.

[0118] In some embodiments, the polymer units are TIFF2026503087000096.tif13128 or a salt thereof.

[0119] In some embodiments, R 4 and R 5 is independently selected from H and a chelator, wherein the chelator is selected from —NR by alkylene, arylene, carbocyclyl, heteroarylene, or heterocarbocyclyl; 4 R 5 may be attached to the nitrogen of R 4 and R 5 and n are not both H.

[0120] In some embodiments, linker compounds are provided 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.

[0121] In some embodiments, the following: A linker compound is provided, comprising a polar group selected from TIFF2026503087000097.tif83130 or a stereoisomer or salt thereof; where the wavy lines are bonds to the amino acid units.

[0122] In some embodiments, R 4 and R 5 are independently selected from H, a polyhydroxyl ether group, and a substituted polyhydroxyl ether group.

[0123] In some embodiments, —(NR 4 R 5 ) but the following: A linker compound is provided, selected from TIFF2026503087000098.tif25128 or a stereoisomer or salt thereof.

[0124] In some embodiments, the following: (a) ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -R 3 (XXX) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or a C1-C3 alkylene group; R 3 is an optionally substituted C3 to C 10 or R is selected from a carbocycle, a thiourea, an optionally substituted thiourea, a urea, an optionally substituted urea, a sulfamide, an alkylsulfamide, an acylsulfamide, an optionally substituted alkylsulfamide, an optionally substituted acylsulfamide, a sulfonamide, an optionally substituted sulfonamide, a guanidine including an alkylguanidine and an arylguanidine, a phosphoramide, or an optionally substituted phosphoramide; or R3 represents azido, alkynyl, substituted alkynyl, -NH-C(O)-alkynyl, -NH-C(O)-alkynyl-R 5 , cyclooctyne; -NH-cyclooctyne, -NH-C(O)-cyclooctyne, or -NH-(cyclooctyne)2; where R 5 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; n0 is 2 to 26; (b) ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -NH-C(O)-R 3 (XXXI) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or a C1-C3 alkylene group; R 3 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 4 having; R 4 are azide, alkynyl, and alkynyl-R 5 , cyclooctyne, or cyclooctyne-R 5 where R 5 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; n0 is 2 to 26; (c) ~R 0 -R1 -[O-CH2-CH2] n0 -R 2 -C(O)NH-R 3 (XXXII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or a C1-C3 alkylene group; R 3 is a branched polyethylene glycol chain, each branch independently having 1 to 26 ethylene glycol subunits and each branch having R at its end 4 having; R 4 are azide, alkynyl, and alkynyl-R 5 , cyclooctyne, or cyclooctyne-R 5 where R 5 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; n0 is 2 to 26; (d) ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -C(O)NR 3 -R 2 -NR 4 R 5 (XXXIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 3 is H or R 2 -NR 4 R 5 and; R 1 and R 2each independently represents a bond or a C1-C3 alkylene group; R 4 and R 5 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, where R 4 and R 5 Both of them cannot be H; n0 is 2 to 26; (e) ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -NR 6 -R 3 (XXXIV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or a C1-C3 alkylene group; R 3 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 4 having; R 6 is C1-C3 alkylene, C1-C3 alkylene-C(O), -C(O)-C1-C3 alkylene, or -C(O)-C1-C3 alkylene-C(O); R 4 are azide, alkynyl, and alkynyl-R 5 , cyclooctyne, or cyclooctyne-R 5 where R 5 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; n0 is 2 to 26; (f) ~R 0 -(R 1 -[CH2-CH(OR 3 )-CH2-O] n0 -R 6 ) n2 (XXXV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 are independently a bond, —O—, or a C1-C3 alkylene group; Each R 3 are independently H, -[CH2-CH(OH)-CH2-O] n0 -R 6 , -C(O)-NR 4 R 5 or -C(O)N(R N )-C1~C6 alkylene-NR 4 R 5 and; R N is H or C1-C4 alkyl; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, where R 4 and R 5 Both of them cannot be H; Each R 6 are independently H, C1-C6 alkylene -C(OH)H-NR 7 R 8 , C1-C6 alkylene-C(OH)H-C1-C6 alkylene-NR 7 R 8 , -C(O)-NR 4 R 5 , -C(O)N(R N )-C1~C6 alkylene-NR 4 R 5 , C1-C6 alkylene-C(O)NR 4 R 5 or C1-C6 alkylene-CO2R 9 and; Each R 9are independently H or C1-C6 alkyl; R 7 and R 8 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; each n0 is independently 1 to 26; n2 is 1 or 2; (g) ~R 0 -R 1 -[[CH2-CH2-O] n0 -R 2 -[CH2-[CH(OH)] n3 -CH2-O] n1 ] n2 -R 3 -NR 4 -R 5 (XXXVI) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 , R 2 and R 3 each independently represents a bond or a C1-C3 alkylene group; R 4 and R 5 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, where R 4 and R 5 Both of them cannot be H; each n0 independently ranges from 0 to 26 and each n1 independently ranges from 0 to 26, provided that at least one of n0 or n1 ranges from 2 to 26; n2 is 1 to 5; each n3 is independently 1 or 2; (h) ~R 0 -(R 1 -[O-CH2-CH2] n0 -R 2 -N(RN )-CO2-[CH2-CH(OR 3 )-CH2-O] n1 -R 6 ) n5 (XXXVII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or a C1-C3 alkylene group; R N is H or C1-C4 alkyl; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, where R 4 and R 5 Both of them cannot be H; Each R 3 are independently H, -[CH2-CH(OH)-CH2-O] n0 -R 6 or -C(O)N(R N )-C1~C6 alkylene-NR 4 R 5 and; Each R 6 are independently H, C1-C6 alkylene -C(OH)H-NR 7 R 8 , C1-C6 alkylene-C(OH)H-C1-C6 alkylene-NR 7 R 8 , -C(O)N(R N )-C1~C6 alkylene-NR 4 R 5 , C1-C6 alkylene-C(O)NR 4 R 5 or C1-C6 alkylene-CO2R 9 and; Each R 9 are independently H or C1-C6 alkyl; R 7 and R 8are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, and a substituted —C(O)-polyhydroxyl group; n0 is 2 to 26; n1 is 1 to 26; n5 is 1 or 2; (i) ~R 0 -(R 1 -[N(R N )-C(O)-[O-CH2-CH(OH)-CH2] n0 ] n1 -R 2 -NR 4 R 5 ) n5 (XXXVIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or a C1-C3 alkylene group; R N is H or C1-C4 alkyl; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, where R 4 and R 5 Both of them cannot be H; n0 is 2 to 26; n1 is 2 to 4; n5 is 1, 2 or 3; (j) ~R 0 -(R 1 -[C(R α )HC(O)-N(R N )] n0 -R 2 -[CH2-CH2-O] n0 -NR 4 R 5 ) n5 (XXXIX) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond, C1-C3 alkylene, -C1-C3 alkylene-[O-CH2-CH2-] n0 , -[CH2-CH2-O] n0 -C1-C3 alkylene-, or -C1-C3 alkylene-[O-CH2-CH2-] n0 -C(O)-; Each R α are independently H or -R 2 -NR 4 R 5 and; Each R N are independently H, C1-C6 alkyl, or -R 2 -NR 4 R 5 and; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a chelating agent, and —C(O)—R, where R is a saccharide unit of formula (XII) or (XIII); or —NR 4 R 5 together form a C3-C8 heterocycle, and R 4 and R 5 Both of them cannot be H; Each n0 is independently 0 to 26, provided that at least one n 0 is 2 to 26; n5 is 1 or 2; or (k) TIFF2026503087000099.tif32128 or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 , R 2 and R 3each independently represents a bond, C1-C3 alkylene, -C1-C3 alkylene-[O-CH2-CH2-] n0 , -[CH2-CH2-O] n0 -C1-C3 alkylene-, or -C1-C3 alkylene-[O-CH2-CH2-] n0 -C(O)-; Each R α are independently H or -R 2 -NR 4 R 5 and; Each R N are independently H, C1-C6 alkyl, or -R 2 -NR 4 R 5 and; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a chelating agent, —C(O)—R, where R is a saccharide unit of formula (XII) or (XIII); or —NR 4 R 5 together form a C3-C8 heterocycle, and R 4 and R 5 Both of them cannot be H; R 6 is H or C1-C6 alkyl; each n0 independently ranges from 0 to 26, provided that at least one n0 ranges from 2 to 26; each n1 independently is 0 to 26, provided that at least one n1 is 2 to 26; Linker compounds are provided that include a polar group having a formula selected from:

[0125] In some embodiments, the following: ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -NH-C(O)-R 3 (XXXI), ~R 0 -R 1-[O-CH2-CH2] n0 -R 2 -C(O)NH-R 3 (XXXII), and ~R 0 -R 1 -[O-CH2-CH2] n0 -R 2 -N-(R 6 -R 3 )2(XXXIII); A linker compound is provided which includes a polar group having a formula selected from: During the ceremony, R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or a C1-C3 alkylene group; R 3 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 4 having; R 6 is C1-C3 alkylene, -C1-C3 alkylene-C(O), -C(O)-C1-C3 alkylene, or -C(O)-C1-C3 alkylene-C(O); R 4 are azide, alkynyl, and alkynyl-R 5 , cyclooctyne, or cyclooctyne-R 5 where R 5 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 0 indicates the binding site of the amino acid unit to; n0 is 2 to 26.

[0126] In some embodiments, the following: a linker compound is provided, comprising a polar group formed from a precursor group selected from TIFF2026503087000100.tif219132; 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 is the bond to the amino acid unit.

[0127] In some embodiments, the formula: ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 (XL) or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C6 alkylene; Each R 3 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 7 R 8 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 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 alkylene, one of which is C1-C2 alkylene, and the nitrogen atom is NR 44 R 45 It is bound to; R 4 and R 5 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, and wherein R 4 and R 5 Both of them cannot be H; n0 is 2 to 26; n1 is 1 to 6; n2 is 1 to 6.

[0128] In some embodiments, the formula: ~R 0 -(R 1 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 )n2 (XLI) or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C6 alkylene; Each R 3 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 7 R 8 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 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12alkylene, one of which is C1-C2 alkylene, and the nitrogen atom is NR 44 R 45 It is bound to; R 4 and R 5 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, and wherein R 4 and R 5 Both of them cannot be H; n0 is 2 to 26; n1 is 1 to 6; n2 is 1 to 6.

[0129] In some embodiments, the formula: ~R 0 -(R 1 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 (XLII) or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C3 alkylene; Each R 3 are independently 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 7 R 8 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 7 and R 8 One of the groups is H or C1-C6 alkylene, and the other is C1-C 12 alkylene, one of which is C1-C2 alkylene, and the nitrogen atom is NR 44 R 45 It is bound to; R 4 and R 5 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, and wherein R 4 and R 5 Both of them cannot be H; n0 is 2 to 16; n1 is 1 to 4; n2 is 1 to 4.

[0130] In some embodiments, R 0is derived from a functional group of a precursor compound to the polymer unit, 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 protected forms thereof.

[0131] In some embodiments, R 0 But the following structure: TIFF2026503087000101.tif175162TIFF2026503087000102.tif157154 or a stereoisomer thereof; where R is H, C1-C6 alkyl or polyhydroxyl group, n is 0-12, and (*) indicates the R 0 The binding sites of each TIFF2026503087000103.tif4128 is the R to the rest of the polymer units 0 The binding site of

[0132] In some embodiments, R 0 But the following structure: TIFF2026503087000104.tif44148TIFF2026503087000105.tif191161TIFF2026503087000106.tif107153 or a stereoisomer thereof; where R is H, C1-C6 alkyl or polyhydroxyl group, n is 0-12, and (*) indicates the R 0 The binding sites of each TIFF2026503087000107.tif4128 is the R to the rest of the polymer units 0 The binding site of

[0133] In some embodiments, R3 If present, -R 3 -(NR 4 R 5 ) n1 But the following structure: TIFF2026503087000108.tif86160TIFF2026503087000109.tif18128 or a stereoisomer thereof; In the formula, each R a and R b are independently H or C 1~6 is alkyl, and X 4 is SO3H, p is 0 to 8, TIFF2026503087000110.tif4128 is the R to the rest of the polymer units 3 The binding site of

[0134] In some embodiments, R 3 If present, -R 3 -(NR 4 R 5 ) n1 has the following structure: TIFF2026503087000111.tif115160 or a stereoisomer thereof, During the ceremony, TIFF2026503087000112.tif4128 is the R to the rest of the polymer units 3 The binding site of

[0135] In some embodiments, at least one -NR 4 R 5 If present, the following structure: TIFF2026503087000113.tif154161TIFF2026503087000114.tif141153 or a stereoisomer thereof; During the ceremony, TIFF2026503087000115.tif4128 -NR to the rest of the polymer units 4 R 5The binding site of

[0136] In some embodiments, the linker unit has the following structure prior to attachment: TIFF2026503087000116.tif57128TIFF2026503087000117.tif231156TIFF2026503087000118.tif238102TIFF20265 03087000119.tif214115TIFF2026503087000120.tif208164TIFF2026503087000121.tif225162TIFF20265030870001 22.tif230168TIFF2026503087000123.tif188162TIFF2026503087000124.tif188162TIFF2026503087000125.tif20 5162TIFF2026503087000126.tif225163TIFF2026503087000127.tif220141TIFF2026503087000128.tif191170TIFF2 026503087000129.tif221161TIFF2026503087000130.tif226169TIFF2026503087000131.tif243166TIFF202650308 7000132.tif183165TIFF2026503087000133.tif203145TIFF2026503087000134.tif220167TIFF2026503087000135.t if228168TIFF2026503087000136.tif237106TIFF2026503087000137.tif20492TIFF2026503087000138.tif244170TIFF2026503087000139.tif222170TIFF2026503087000140.tif190170, or a stereoisomer thereof; During the ceremony, (*) indicates the binding site to the amino acid unit; Each R, R a and R b are independently H or C1-C6 alkyl; R' is H, C1-C6 alkyl, -N(R 4 )(R 5 ) or -CO2H; each n is independently 1 to 12; X is O, NR, or -CH2-; V is a bond or C1-C6 alkyl; R 4 and R 5 is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, or -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII); and R 4 and R 5 the other is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a chelating agent, or —C(O)—R, where R is a saccharide unit of formula (XII) or (XIII), and polyethylene glycol optionally having 1 to 24 ethylene glycol subunits; or —NR 4 R 5 together form a C3-C8 heterocycle, and R 4 and R 5 Both of them cannot be H.

[0137] In some embodiments, the following: (a) ~R 0 -(R 3 -R 1 -[O-CH2-CH2] n0 -R 6 -[O-CH2-CH2] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 (XLIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 each independently represents a bond or C1-C6 alkylene; Each R 3 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)-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C1~C 12 alkylene, and the other is C1-C 12 is alkylene; R 4 and R 5 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, and wherein R 4 and R 5 Both of them cannot be H; Each R 6 are independently, -NR a -, -NR a -C1~C6 alkylene-NR a-, -NR a -C(O)-NR a -S(O)2-NR a -, or -NR a -C(O)-C 1~6 alkylene-selected from; Each R a are independently selected from H, C1-C6 alkyl, or polyhydroxyl groups; each n0 is independently 2 to 26; n1 is 1 to 6; n2 is 1 to 6; (b) ~R 0 -(R 1 -[O-CH2-CH2] n3 -R 2 -X1-R 5 -X2-R 3 -[O-CH2-CH2] n3 -R 4 -[X3-R 6 ] n4 -R 7 ) n5 (XLIV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 , R 2 , R 3 and R 4 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 5 and R 6 each independently represents a divalent polyhydroxyl group; R 7 is H, OH or C1-C6 alkyl; each n3 independently is 0 to 26, provided that at least one n3 is 2 to 26; n4 is 0 to 10; n5 is 1 or 2; or (c) ~R 0 -R 1 -[O-CH2-CH2] n3 -R 2 -N-(R 3 -X1-R 4 -[O-CH2-CH2] n3 -(NR 4 R 5 ))2 (XLV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 , R 3 and R 4 each independently represents an optionally bonded or substituted C1-C6 alkylene; Each R 2 are independently 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 4 and R 5 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, and wherein R 4 and R 5 Both of them cannot be H; each n3 is independently 2 to 26; Linker compounds are provided that include a polar group having a formula selected from:

[0138] In some embodiments, the amino acid unit has the following structure prior to attachment: TIFF2026503087000141.tif101128TIFF2026503087000142.tif165170, During the ceremony, (*) indicates the binding site to the amino acid unit; Each R a are independently H, alkyl, or polyhydroxyl groups; R 4 and R 5 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, and wherein R 4 and R 5 Both of them cannot be H; Each n is independently 1 to 12.

[0139] In some embodiments, TIFF2026503087000143.tif212157 or a stereoisomer or salt thereof, During the ceremony, Each Y is independently R 76 or TIFF2026503087000144.tif19128; 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 a bond to an amino acid unit.

[0140] In some embodiments, TIFF2026503087000145.tif223164 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 a bond to an amino acid unit.

[0141] In some embodiments, TIFF2026503087000146.tif58128TIFF2026503087000147.tif160166 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 a bond to an amino acid unit.

[0142] In some embodiments, Y is R 76 A linker compound is provided, wherein:

[0143] In some embodiments, Y is A linker compound is provided, which is TIFF2026503087000148.tif19128.

[0144] In some embodiments, each R a and R b are independently H.

[0145] In some embodiments, R a and R b taken together with the carbon to which they are attached form an oxo group.

[0146] In some embodiments, linker compounds are provided wherein q is 10-20.

[0147] In some embodiments, linker compounds are provided wherein q is 12.

[0148] In some embodiments, the polar group has the following structure prior to attachment to the amino acid unit: TIFF2026503087000149.tif173148TIFF2026503087000150.tif243144TIFF2026503087000151.tif155141 or a stereoisomer thereof; In the formula, R a is H or C 1~6 alkyl, and n is 1 to 20.

[0149] In some embodiments, the polar group has the following structure prior to attachment to the amino acid unit: TIFF2026503087000152.tif228138TIFF2026503087000153.tif193170TIFF2026503087000154.tif249165TIFF2026503087000155.tif231141TIFF2026503087000156.tif213149TIFF2026503087000157.tif50128, or a stereoisomer thereof; In the formula, R a is H or C 1~6 alkyl, and n is 1 to 20.

[0150] In some embodiments, the polar group has the following structure prior to attachment to the amino acid unit: TIFF2026503087000158.tif130138TIFF2026503087000159.tif234146TIFF2026503087000160.tif239170TIFF2026503087000161.tif247150TIFF2026503087000162.tif178146TIFF2026503087000163.tif129128, or a stereoisomer thereof; In the formula, R a is H or C 1~6 alkyl, and n is 1 to 20.

[0151] In some embodiments, the following: TIFF2026503087000164.tif59128TIFF2026503087000165.tif218159TIFF2026503087000166.tif210158TIFF2026503087000167.tif197158TI FF2026503087000168.tif209158TIFF2026503087000169.tif205158TIFF2026503087000170.tif222158TIFF2026503087000171.tif212158TIF F2026503087000172.tif181156TIFF2026503087000173.tif245158TIFF2026503087000174.tif243159TIFF2026503087000175.tif215159TIFF 2026503087000176.tif199158TIFF2026503087000177.tif207156TIFF2026503087000178.tif205156TIFF2026503087000179.tif207158TIFF20 26503087000180.tif187148TIFF2026503087000181.tif226146TIFF2026503087000182.tif238146TIFF2026503087000183.tif238143TIFF202 6503087000184.tif213149TIFF2026503087000185.tif232138TIFF2026503087000186.tif205146TIFF2026503087000187.tif238120TIFF2026 503087000188.tif22080TIFF2026503087000189.tif235158TIFF2026503087000190.tif192159TIFF2026503087000191.tif238159TIFF2026503087000192.tif192160TIFF2026503087000193.tif207160TIFF2026503087000194.tif202163, or a stereoisomer or salt thereof; wherein each Z is attached at *; TIFF2026503087000195.tif31148TIFF2026503087000196.tif90159 are independently selected, TIFF2026503087000197.tif2128 is the bond to the amino acid unit.

[0152] In some embodiments, the following: a linker compound comprising a polar group selected from TIFF2026503087000198.tif123136TIFF2026503087000199.tif233170TIFF2026503087000200.tif240161TIFF2026503087000201.tif237164TIFF2026503087000202.tif222164, or a stereoisomer or salt thereof; During the ceremony, each TIFF2026503087000203.tif2128 shows the binding to amino acid units.

[0153] In some embodiments, the polar group has the formula: A linker compound is provided, comprising at least one carboxyl unit having TIFF2026503087000204.tif38128 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)-, where * is the bond to the amino acid unit or to the remainder of the polar group; R 70 Ha~NR 71 (R 72 -R 73 ), where R 71 H, C1~C 12 Alkyl, substituted C1-C 12alkyl, 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 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; (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)-, where * is the bond to the amino acid unit or to the remainder of the polar group; R 70 Ha~NR 71 (R 75 - (R 73 )2), where R 71 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 R73 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; or (c) 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)-, where * is the bond to the amino acid unit 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.

[0154] In some aspects, a linker compound is provided that includes a polar group that includes a polymer unit and a sugar unit.

[0155] In some embodiments, a linker compound is provided that includes a polar group comprising at least two polymer units.

[0156] In some aspects, a linker compound is provided that includes a polymeric unit and a polar group that includes a carboxyl unit.

[0157] In some embodiments, a linker compound is provided that includes at least two polar groups.

[0158] In some aspects, a linker compound is provided that includes a polymer unit, a saccharide unit, and a polar group that includes a carboxyl unit.

[0159] In some embodiments, a linker compound is provided that includes a polar group that includes at least two polymer units, at least one sugar unit, and at least one carboxyl unit.

[0160] In some embodiments, linker compounds are provided wherein the amino acid unit comprises at least two amino acid subunits.

[0161] In some embodiments, a linker compound is provided that, when present, comprises two polar groups, both attached to an amino acid unit.

[0162] In some aspects, linker compounds are provided in which the linker unit is attached to a side chain of a subunit of an amino acid unit.

[0163] In some aspects, linker compounds are provided in which the amino acid units are joined to the linker units by non-peptide linking groups.

[0164] 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 A linker compound is provided that is selected from alkynylene, or optionally substituted polyethylene glycol.

[0165] In some embodiments, the following structure: TIFF2026503087000205.tif250158TIFF2026503087000206.tif48130 or a stereoisomer thereof, wherein a polar group is attached to an amino acid subunit of the Amino Acid Unit, and an H of the hydroxyl or amino group of the para-aminobenzyl group or an H of the hydroxyl of the glycine residue of the GGFG peptide may be replaced with a bond to at least one of the Drug Units or to a linking group attached to at least one of the Drug Units, and a wavy line on the amino group indicates the attachment site for a Stretcher Unit or an Amino Acid Unit, or indicates an H prior to attachment. In some embodiments, at least one of the Drug Units is attached directly to the benzyl oxygen (—O—). In some embodiments, at least one of the Drug Units is attached indirectly via a linking group. The linking group may be any suitable group for attaching at least one drug unit to benzyl-O-, which 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-CH2-CH2-CH2-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 another embodiment, the linking group is -C(O)-NH-CH2-CH2-CH2-C(O)-.

[0166] In some embodiments, the following: A linker compound is provided, comprising a formula selected from TIFF2026503087000207.tif31128, In the formula, square brackets indicate amino acid units, each aa is any subunit of an amino acid unit, L2 is a linker unit, and each wavy line (~) indicates a binding site for a Stretcher unit; aa1 (poly) is a polymer unit attached to an amino acid subunit of an amino acid unit, SU is a sugar unit attached to a subunit of an amino acid unit or to a linker unit, and CU is a carboxyl unit attached to a subunit of an amino acid unit or to a linker unit; a double wavy line TIFF2026503087000208.tif4128 shows a binding site for at least one of the Drug units, where aa and aa1 are independently selected from α, β and γ amino acids and derivatives thereof.

[0167] In some embodiments, the following: A linker compound is provided, comprising a formula selected from TIFF2026503087000209.tif70128, In the formula, the square brackets indicate amino acid units, each aa is an amino acid subunit of the amino acid unit, L2 is a linker subunit attached to the side chain of aa, the wavy line (~) indicates the attachment site for the Stretcher unit; aa1 (poly) is a polymer unit attached to aa, SU is a sugar unit attached to aa, CU is a carboxyl unit attached to aa, and the double wavy line TIFF2026503087000210.tif4128 shows a binding site for at least one of the Drug units, where aa and aa1 are independently selected from α, β and γ amino acids and derivatives thereof.

[0168] In some embodiments, linker compounds are provided in which at least two polymer units are attached to an amino acid unit.

[0169] In some embodiments, the following: A linker compound is provided, comprising a formula selected from TIFF2026503087000211.tif31128, In the formula, the square brackets indicate amino acid units, aa is any subunit of an amino acid unit, L2 is a linker unit, the wavy line (~) indicates the attachment site for a Stretcher unit; aa1 (poly) and aa2 (poly) are polymer units attached to aa or other polymer units, respectively, each SU is a sugar unit attached to aa or other sugar unit, each CU is a carboxyl unit attached to aa or other carboxyl unit, and the double wavy line TIFF2026503087000212.tif4128 shows a binding site for at least one of the Drug units; where aa, aa1 and aa2 are independently selected from α, β and γ amino acids and derivatives thereof.

[0170] In some embodiments, the following: A linker compound is provided, comprising a formula selected from TIFF2026503087000213.tif70128, In the formula, the square brackets indicate amino acid units, aa is an amino acid subunit of the amino acid unit, L2 is a linker unit attached to the side chain of aa, each wavy line (~) indicates a binding site for a Stretcher unit; aa1 (poly) and aa2 (poly) are each polymer units attached to aa, each SU is a saccharide unit attached to aa; each CU is a carboxyl unit attached to aa; and double wavy lines TIFF2026503087000214.tif4128 shows a binding site for at least one of the Drug units; where each of aa, aa1 and aa2 is independently selected from α, β and γ amino acids and derivatives thereof.

[0171] In some aspects, linker compounds are provided wherein the linker unit is a cleavable linker unit.

[0172] In some aspects, linker compounds are provided in which the linker unit comprises a peptide that is cleavable by an intracellular protease.

[0173] In some embodiments, the intracellular protease is cathepsin B.

[0174] In some embodiments, linker compounds are provided wherein 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.

[0175] In some aspects, linker compounds are provided in which the amino acid units comprise peptides that are cleavable by intracellular proteases.

[0176] In some embodiments, linker compounds are provided wherein 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.

[0177] In some embodiments, linker compounds are provided in which a cleavable peptide is attached to a para-aminobenzyl alcohol self-immolative group (PABA).

[0178] In some embodiments, the following structure: TIFF2026503087000215.tif174138TIFF2026503087000216.tif231146TIFF2026503087000217.tif237163TIFF2026503087000218.tif237151TIFF2026503087000219.tif250146TIFF2026503087000220.tif195146TIFF2026503087000221.tif199163TIFF2026503087000222.tif226159; In the formula, the wavy line on the oxygen group TIFF2026503087000223.tif2128 or *-amino group indicates the site of attachment to at least one of the Drug units or for a linking group attached to at least one of the Drug units; wavy line on amino group TIFF2026503087000224.tif2128 indicates the binding site for the Stretcher unit or amino acid unit, or H before binding.

[0179] In some embodiments, at least one of the drug units is directly bonded to the benzyl O. In some embodiments, at least one of the drug units is indirectly bonded via a linking group. The linking group may be any suitable group for bonding at least one drug unit to the benzyl oxygen (—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).

[0180] In some aspects, linker compounds are provided wherein the linker unit further comprises a stretcher unit having a binding site for a targeting unit, wherein the stretcher unit is attached to an amino acid unit of the linker compound.

[0181] In some embodiments, the stretcher unit comprises: Selected from TIFF2026503087000225.tif99144, wherein: each TIFF2026503087000226.tif4128 shows the binding sites to amino acid units; 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 10Alkylene-(C3-C8 carbocyclo)-, -(C3-C8 carbocyclo)-C1-C 10 Alkylene-, -C3-C8 heterocyclo-, -C1-C 10 Alkylene-(C3-C8 heterocyclo)-, -(C3-C8 heterocyclo)-C1-C 10 Alkylene-, -C1~C 10 Alkylene-C(=O)-, C1-C 10 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 The stretcher unit is maleimide (C1-C 10 Alkylene-C(O)-, maleimide (CH2OCH2) p2 (C1~C 10 Alkylene)C(O)-, Maleimide (C1-C 10 alkylene) (CH2OCH2) p2 C(O)- or their open ring forms, wherein p2 is 1 to 26; A linker compound is provided.

[0182] In some embodiments, the stretcher unit comprises: a linker compound selected from TIFF2026503087000227.tif83140TIFF2026503087000228.tif43144 or a stereoisomer thereof; In the formula, each R a are independently H or C 1~6 alkyl, each n is independently 0 to 12, TIFF2026503087000229.tif2128 shows the attachment site of the Stretcher unit to the Amino Acid unit, and the attachment site for the Targeting unit is on the maleimide, primary amine, or alkyne functional group.

[0183] In some embodiments, the stretcher unit comprises: a linker compound selected from TIFF2026503087000230.tif138143TIFF2026503087000231.tif40153 or a stereoisomer thereof; In the formula, wavy line TIFF2026503087000232.tif2128 shows the attachment sites of the Stretcher unit to the Amino Acid unit, and the attachment sites for the Targeting unit are on the maleimide, primary amine, or alkyne functional group.

[0184] In some embodiments, the following structure: TIFF2026503087000233.tif134128TIFF2026503087000234.tif224164TIFF2026503087000235.tif250164TIFF2026503087000236.tif217164TIFF2026503087000237.tif234165TIFF2026503087000238.tif198164TIFF2026503087000239.tif231164TIFF2026503087000240.tif200164, or a stereoisomer thereof; In the formula, wavy line TIFF2026503087000241.tif2128 represents the attachment site to at least one of the Drug units or for a linking group attached to at least one of the Drug units.

[0185] In some embodiments, a Drug-Linker Compound is provided that comprises a linker compound described herein conjugated to at least one Drug unit.

[0186] 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.

[0187] In some embodiments, Drug-Linkers are provided wherein the Drug unit is a cytotoxic agent.

[0188] In some embodiments, a drug-linker is provided wherein the cytotoxic agent is selected from the group consisting of an auristatin, a maytansinoid, a camptothecin, a duocarmycin, and a calicheamicin.

[0189] In some embodiments, a drug-linker is provided wherein the cytotoxic agent is an auristatin.

[0190] In some embodiments, a drug-linker is provided wherein the cytotoxic agent is MMAE or MMAF.

[0191] In some embodiments, a drug-linker is provided wherein the cytotoxic agent is camptothecin.

[0192] In some embodiments, a drug-linker is provided wherein the cytotoxic agent is exatecan or SN-38.

[0193] In some embodiments, a drug-linker is provided wherein the cytotoxic agent is RS-exatecan or SS-exatecan.

[0194] In some embodiments, a drug-linker is provided wherein the cytotoxic agent is calicheamicin.

[0195] In some embodiments, a Drug-Linker is provided wherein the cytotoxic agent is a maytansinoid.

[0196] In some embodiments, a Drug-Linker is provided wherein the maytansinoid is maytansine, maytansinol, or ansamatocin-2.

[0197] In some embodiments, a Drug-Linker is provided wherein the Drug unit is an immunomodulator.

[0198] In some embodiments, a drug-linker is provided wherein the immunomodulator is selected from a TRL7 agonist, a TLR8 agonist, a STING agonist, or a RIG-I agonist.

[0199] In some embodiments, a drug-linker is provided wherein the immunomodulator is a TLR7 agonist.

[0200] In some embodiments, a drug-linker is provided 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.

[0201] In some embodiments, a Drug-Linker is provided wherein the immunomodulator is a TLR8 agonist.

[0202] In some embodiments, a drug-linker is provided wherein the TLR8 agonist is selected from imidazoquinoline, thiazoloquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzimidazol-2-amine, tetrahydropyridopyrimidine, or ssRNA.

[0203] In some embodiments, a drug-linker is provided wherein the immunomodulator is a STING agonist.

[0204] In some embodiments, a drug-linker is provided wherein the immunomodulator is a RIG-I agonist.

[0205] In some embodiments, a drug-linker is provided wherein the RIG-I agonist is selected from KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400, and KIN2000.

[0206] In some embodiments, a Drug-Linker is provided wherein the Drug unit is a chelating ligand.

[0207] In some embodiments, a drug-linker is provided 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.

[0208] In some embodiments, the following structure: TIFF2026503087000242.tif153164TIFF2026503087000243.tif191164TIFF2026503087000244.ti f179164TIFF2026503087000245.tif251164TIFF2026503087000246.tif223164TIFF2026503087000 247.tif240165TIFF2026503087000248.tif184165TIFF2026503087000249.tif204164TIFF2026503087000250.tif195164TIFF2026503087000251.tif61144 or stereoisomers thereof.

[0209] In some embodiments, a conjugate is provided that includes a targeting unit attached to a drug-linker described herein, wherein the targeting unit specifically binds to a target molecule.

[0210] In some aspects, a conjugate is provided wherein the targeting unit is selected from an antibody or an antigen-binding portion thereof.

[0211] In some embodiments, conjugates are provided wherein the targeting unit is a monoclonal antibody, a Fab, a Fab', a F(ab'), an Fv, a disulfide-linked Fc, an scFv, a single domain antibody, a diabody, a bispecific antibody, or a multispecific antibody.

[0212] In some embodiments, conjugates are provided wherein the targeting unit 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.

[0213] In some embodiments, conjugates are provided wherein the targeting unit is a diabody, DART, anticalin, affibody, avimer, DARPin, or adnectin.

[0214] In some embodiments, conjugates are provided in which the targeting unit is monospecific.

[0215] In some embodiments, conjugates are provided wherein the targeting unit is bivalent.

[0216] In some embodiments, conjugates are provided wherein the targeting unit is bispecific.

[0217] In some embodiments, 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.

[0218] In some embodiments, the following: TIFF2026503087000252.tif186164TIFF2026503087000253.tif239164TIFF2026503087 000254.tif225164TIFF2026503087000255.tif251164TIFF2026503087000256.tif2211 64TIFF2026503087000257.tif200164TIFF2026503087000258.tif202164TIFF2026503087000259.tif198164TIFF2026503087000260.tif128160 or a stereoisomer thereof.

[0219] In some embodiments, a conjugate is provided, wherein the targeting unit binds to a target molecule.

[0220] In some embodiments, conjugates are provided wherein the target molecule is CD19, CD20, CD30, CD33, CD70, LIV-1, HER2, or EGFRv3. In some embodiments, conjugates are provided wherein the target molecule is CD19, CD20, CD30, CD33, CD70, LIV-1, or EGFRv3.

[0221] In some embodiments, a conjugate is provided wherein the target molecule is a cancer-associated antigen.

[0222] In some embodiments, the target molecule is CD19, CD20, CD30, CD33, CD38, CA125, 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), HER2, 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, TUBB 3, VHL, CDH1, ERBB4, FBXW7, HNF1A, JAK3, NPM1, PTPN11, RB1, SMAD4, SMARCB1, STK1, MLH1, MSH2, MSH6, PMS2, ROS1, ERCC1, 5T4(TPBG), B7-H3, CCR7, CD 105, 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.

[0223] In some embodiments, the targeting unit 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 are provided that are antibodies, or fragments thereof, 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 anti-LHRH receptor antibodies, including clones A9E4, F1G4, AT2G7, GNRH03, or GNRHR2.

[0224] In some aspects, a pharmaceutical composition is provided comprising a conjugate described herein and a pharmaceutically acceptable carrier.

[0225] 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.

[0226] Sugar Unit (SU) In some embodiments, the sugar unit (SU) has the general formula (X): L3-N(CH2-(CH(X1R)) k -X2(X3))2(X) or a stereoisomer or salt thereof, During the ceremony, each X1 is independently selected from NH or O; each R is independently selected from hydrogen, acetyl, monosaccharides, disaccharides, and polysaccharides; each X2 is independently selected from CH2 and C(O); each X3 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.

[0227] In some embodiments, the sugar unit has the following structure (XII) or (XIII): TIFF2026503087000261.tif87128 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.

[0228] Carboxyl Unit In some embodiments, the linker comprises a carboxyl unit. In some embodiments, the carboxyl unit has the following general formula (XXXX): TIFF2026503087000262.tif38128 or a stereoisomer or salt thereof, wherein: (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)-, where * is the bond to the amino acid unit or to the remainder of the polar group; R 70 Ha~NR 71 (R 72 -R 73 ), where R 71 H, C1~C 12 Alkyl, substituted C1-C 12 alkyl, or polyethylene glycol (which may have 1 to 12 ethylene glycol subunits); R72 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 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; (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)-, where * is the bond to the amino acid unit or to the remainder of the polar group; R 70 Ha~NR 71 (R 75 - (R 73 )2), where R 71 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 73are 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; or (c) 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)-, where * is the bond to the amino acid unit 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.

[0229] Linker Unit As further described herein, the linker comprises at least one linker unit or linker subunit L2, and each linker unit or linker subunit L2 has a binding site for at least one drug unit (D). In some embodiments, a drug unit (D) is attached to each binding site for a drug unit on the linker unit or linker subunit L2. In various embodiments, the linker unit or linker subunit L2 may be a cleavable linker unit or a non-cleavable linker unit. The linker unit or linker subunit L2 also has a binding site for an amino acid unit (AA) or a stretcher unit (L1).

[0230] In some embodiments, the attachment site for the drug unit comprises a linking group. The linking group may be any suitable group for attachment of at least one drug unit that allows for release of the active drug unit or release of 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).

[0231] 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).

[0232] In some embodiments, the linker unit or linker subunit L2 comprises a polar group, such as a saccharide unit, a polymer unit, or a carboxyl unit. In some embodiments, the linker unit or linker subunit L2 does not comprise a polar group, where the amino acid unit comprises a polar group. In some embodiments, both the linker unit or linker subunit L2 and the amino acid unit (if present) comprise a polar group.

[0233] 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 is selected from an optionally substituted polyamide, a substituted polyether, or a combination thereof. In further embodiments, the polymer unit is (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, TIFF2026503087000263.tif19128, 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 TIFF2026503087000264.tif22128, or a stereoisomer thereof; or (iii) any combination thereof is selected from.

[0234] In some embodiments, the linker unit or linker sub-unit L2 is a cleavable linker unit. 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 linker sub-unit L2, or a portion thereof, resulting in a free Drug unit or other metabolite of the linker unit-drug unit or linker sub-unit L2-drug unit dissociated from the remainder of the linker unit or linker sub-unit L2.

[0235] In some embodiments, the linker unit or linker subunit L2 is a protease-cleavable linker unit, an acid-cleavable linker unit, a disulfide linker unit, a disulfide-containing linker unit, or a disulfide-containing linker unit having a dimethyl group adjacent to the disulfide bond (e.g., an SPDB linker) (see, e.g., Jain et al., Pharm. Res. 32:3526-3540 (2015); Chari et al., Cancer Res. 52:127-131 (1992); U.S. Pat. No. 5,208,020)), a cleavable self-stabilizing linker (see, e.g., WO2018 / 031690 and WO2015 / 095755 and Jain et al., Pharm. Res. 32:3526-3540 (2015)) and / or a cleavable hydrophilic linker (see, e.g., WO2015 / 123679). In some embodiments, the linker unit or linker subunit L2 comprises a photolabile linker subunit. In some embodiments, the linker unit or linker subunit L2 has a non-cleavable linker unit (see, e.g., WO2007 / 008603).

[0236] In some embodiments, the linker unit or linker subunit L2 comprises a glucuronide cleavable moiety (see, e.g., US2014 / 0031535).

[0237] In some embodiments, the linker unit or linker subunit L2 is a cleavable linker that is cleavable under intracellular conditions such that cleavage of or within the linker unit or linker subunit L2 releases the Drug unit from the linker unit (or linker subunit L2) or from the remainder of the linker unit within the intracellular environment. For example, in some embodiments, the linker unit or linker subunit L2 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 linker subunit L2, or a portion thereof, resulting in a free Drug unit or other metabolite of the linker unit-drug unit dissociated from the remainder of the linker unit or linker subunit L2 within the cell. The cleaved portion of the conjugate thus becomes an intracellular metabolite. One advantage of using intracellular proteolytic release of the Drug Unit is that the activity of the Drug Unit is typically diminished upon conjugation, and the serum stability of the conjugate is typically increased.

[0238] In some embodiments, the bond between the linker unit or linker subunit L2 and the drug unit can be enzymatically cleaved by one or more enzymes, including tumor-associated proteases, to release the drug unit (D). The linker unit or linker subunit L2 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 (see, e.g., WO2004 / 010957, US20150297748, US2008 / 0166363, US20120328564, and US20200347075). The linker unit or linker subunit L2 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 may include cathepsins 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, e.g., Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123). The peptidyl linker may be cleavable by enzymes present in target antigen-expressing cells. For example, a peptidyl linker subunit (e.g., having a Phe-Leu, Val-Ala, Val-Cit, or Gly-Phe-Leu-Gly peptide) that can be cleaved by cathepsin B, a thiol-dependent protease highly expressed in cancer tissues, may be used.

[0239] Typically, the linker unit or linker subunit L2 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 peptidyl linker is a dipeptide, tripeptide, tetrapeptide, or pentapeptide. In certain embodiments, the peptidyl linker subunit may contain only natural amino acids. In some embodiments, for example, the peptidyl linker subunit may have a Phe-Leu, Val-Ala, Val-Cit, or Gly-Phe-Leu-Gly peptide. Other such cleavable linkers 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 specific embodiments, the peptidyl linker cleavable by intracellular proteases comprises a Val-Cit peptide or a Phe-Lys peptide (see, e.g., U.S. Patent No. 6,214,345) or a Gly-Gly-Phe-Gly linker (see, e.g., U.S. Patent Application Publication No. 2015 / 0297748). One advantage of using intracellular proteolytic release of the drug unit is that the activity of the drug unit is typically reduced upon conjugation, and the serum stability of the conjugate is typically increased. See also U.S. Patent No. 9,345,785.

[0240] In some embodiments, a peptidyl linker subunit can contain only unnatural amino acids. In some embodiments, a peptidyl linker subunit can contain a natural amino acid linked to an unnatural amino acid. In some embodiments, a peptidyl linker subunit can contain a natural amino acid linked to a D-form of a natural amino acid. In some embodiments, at least one amino acid of a peptidyl linker subunit is an L-amino acid. In some embodiments, at least one amino acid is a D-amino acid.

[0241] 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.

[0242] In some embodiments, the peptidyl linker subunits contain one or more of the following: glycine and / or L-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and polar groups (including polymer units attached to glycine or L-amino acids). In some embodiments, the peptidyl linker subunits contain one or more of the following: glycine and / or D-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and polar groups (including polymer units attached to glycine or D-amino acids). In some embodiments, the peptidyl linker subunits contain one or more of the following: glycine and / or a mixture of L- and D-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and a polar group (including a polymer unit attached to glycine or an amino acid).

[0243] In some embodiments, the peptidyl linker subunit contains one or more of the following: glycine and / or natural L-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and at least one polar group, such as a sugar unit, or a carboxyl unit or polymeric unit, attached to the glycine or L-amino acid. In some embodiments, the peptidyl linker subunit contains one or more of the following: glycine and / or a D-amino acid, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and at least one polar group, such as a sugar unit, or a carboxyl unit or polymeric unit, attached to the glycine or D-amino acid.

[0244] In some embodiments, the amino acids of the peptidyl linker subunits have the formula shown below within square brackets: TIFF2026503087000265.tif24128, In the formula, R 190 is 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 TIFF2026503087000266.tif59128.

[0245] In some embodiments, the peptidyl linker subunit 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; and at least one polar group, e.g., a sugar unit, polymer unit, or carboxyl unit, attached to the glycine or natural amino acid.

[0246] In some embodiments, the peptidyl linker subunits do not contain cysteines, hi some embodiments, the peptidyl linker subunits do not contain prolines.

[0247] In some embodiments, the peptidyl linker subunit comprises one or more of the following D-forms of these naturally occurring amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, tryptophan, and valine; and at least one polar group, e.g., a sugar unit, polymer unit, or carboxyl unit, attached to the glycine or D-amino acid.

[0248] In some embodiments, the peptidyl linker subunit 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 acid, aminoalkanoic acid, aminoalkanedioic acid, aminobenzoic acid, amino-heterocyclo-alkanoic acid, heterocyclo-carboxylic acid, citrulline, statin, diaminoalkanoic acid, and derivatives thereof; and at least one polar group attached to the amino acid, e.g., a sugar unit, a polymer unit, or a carboxyl unit. Examples of such amino acid derivatives are described in the section describing the amino acid subunits below.

[0249] In some embodiments, the peptidyl linker subunit contains a saccharide unit as part of the cleavable peptide, such as a saccharide unit containing lysine or citrulline as part of the cleavable peptide. In some embodiments, the peptidyl linker subunit contains a carboxyl unit as part of the cleavable peptide, such as a carboxyl unit containing lysine or citrulline as part of the cleavable peptide.

[0250] In some embodiments, the cleavable linker subunit is pH-sensitive, i.e., sensitive to hydrolysis at a specific pH value. Typically, the pH-sensitive linker subunit is hydrolyzable under acidic conditions. For example, an acid-labile linker subunit (e.g., hydrazone, semicarbazone, thiosemicarbazone, cis-aconitic acid amide, orthoester, acetal, ketal, etc.) that is hydrolyzable in lysosomes can be used. (See, for example, U.S. Patent Nos. 5,122,368, 5,824,805, and 5,622,929; Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123; Neville et al., 1989, Biol. Chem. 264:14653-14661) Such linker subunits are relatively stable under neutral pH conditions, such as in blood, but become unstable below pH 5.5 or 5.0, the approximate pH of the lysosome. In certain embodiments, the hydrolyzable linker unit is a thioether linker (e.g., a thioether attached to the Drug unit via an acylhydrazone bond (see, e.g., U.S. Pat. No. 5,622,929)).

[0251] In some embodiments, the linker unit or linker subunit L2 is cleavable under reducing conditions (eg, a disulfide linker subunit). A variety of disulfide linkers are known, including those that can be formed using, for example, SATA (N-succinimidyl-5-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), and SMPT (N-succinimidyl-oxycarbonyl-α-methyl-α-(2-pyridyl-dithio)toluene)-, SPDB, and SMPT (see, e.g., Thorpe et al., 1987, Cancer Res. 47:5924-5931; Wawrzynczak et al., In Immunoconjugates: Antibody Conjugates in Radioimagery and Therapy of Cancer (C.W. Vogel ed., Oxford U. Press, 1987; see also U.S. Pat. No. 4,880,935).

[0252] In some embodiments, the linker unit or linker subunit L2 is a malonate linker (Johnson et al., 1995, Anticancer Res. 15:1387-93), a maleimidobenzoyl linker (Lau et al., 1995, Bioorg-Med-Chem. 3(10):1299-1304), or a 3'-N-amide analog (Lau et al., 1995, Bioorg-Med-Chem. 3(10):1305-12). In some embodiments, the linker unit or linker subunit L2 is non-cleavable, such as a maleimidocaproyl linker, and the drug unit is released by metabolic degradation of the drug-linker. (See, e.g., U.S. Patent Application Publication No. 2005 / 0238649.)

[0253] In some embodiments, the linker unit or linker subunit L2 is substantially insensitive to the extracellular environment. As used herein, "substantially insensitive to the extracellular environment" in the context of a linker unit or linker subunit L2 means that when the conjugate is present in an extracellular environment (e.g., in plasma), no more than about 20%, typically no more than about 15%, more typically no more than about 10%, and even more typically no more than about 5%, no more than about 3%, or no more than about 1% of the linker units or linker subunits L2 are cleaved in a sample of the conjugate. Whether a linker unit or linker sub-unit L2 is substantially insensitive to the extracellular environment can be determined, for example, by independently incubating both (a) the conjugate (the "conjugate sample") and (b) an equimolar amount of unconjugated targeting unit or drug unit (the "control sample") with plasma for a predetermined period of time (e.g., 2, 4, 8, 16, or 24 hours), and then comparing the amount of unconjugated targeting unit or drug unit present in the conjugated sample with the amount of unconjugated targeting unit or drug unit present in the control sample, as measured, for example, by high performance liquid chromatography.

[0254] In some embodiments, the linker or linker subunit L2 promotes internalization into a cell. In some embodiments, the linker or linker subunit L2 promotes internalization into a cell when conjugated to a drug unit, such as a cytotoxic agent (i.e., in the environment of the linker-drug unit portion of a conjugate described herein). In yet other embodiments, the linker or linker subunit L2 promotes internalization into a cell when conjugated to both a drug unit and a targeting unit (i.e., in the environment of a conjugate described herein).

[0255] Various linker units or linker subunits L2 that can be used in the present compositions and methods are described, for example, in WO2004010957. In some embodiments, the linker unit or linker subunit L2 comprises a protease-cleavable linker comprising a thiol-reactive spacer and a dipeptide (e.g., maleimidyl caproyl valine alanine). In some embodiments, the linker unit or linker subunit L2 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 or linker subunit L2 comprises a protease-cleavable linker comprising a thiol-reactive maleimidocaproyl spacer, a valine-citrulline dipeptide, and a p-amino-benzyloxycarbonyl self-immolative group.

[0256] In some embodiments, the linker unit or linker subunit L2 comprises an acid-cleavable linker such as a hydrazine linker or a quaternary ammonium linker (see, e.g., WO2017 / 096311 and WO2016 / 040684).

[0257] In some embodiments, the linker unit or linker subunit L2 comprises a self-stabilizing moiety comprising a maleimide group as described in WO2013 / 173337.

[0258] In some embodiments, the linker unit or linker subunit L2 comprises a hydrophilic linker, such as, for example, the hydrophilic peptides in WO2015 / 123679, and the sugar alcohol polymer-based linkers disclosed in WO2013 / 012961 and WO2019 / 213046.

[0259] In another embodiment, the linker unit or linker subunit L2 may be made using a variety of bifunctional protein coupling agents, such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxyl (SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCl), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bisazide compounds (such as bis(p-azidobenzoyl)hexanediamine), bisdiazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). Chelating agents for the complexation of radionucleotides are described, for example, in WO 94 / 11026.

[0260] In some embodiments, the linker unit or linker subunit L2 can be prepared using cross-linker reagents including, but not limited to, BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, and sulfo-SMPB, and SVSB (succinimidyl-(4-vinylsulfone)benzoate), which are commercially available (e.g., from Pierce Biotechnology, Inc., Rockford, IL, USA).

[0261] Amino acid (AA) units The linker may comprise an amino acid unit (AA). When present in the linker, the amino acid unit connects the Stretcher unit (L1) to the linker unit. When s in AA is 0, the amino acid unit is absent (e.g., in any of Formulas I-IV). In some embodiments, the amino acid unit comprises 0 to 12 subunits. Each subunit of the amino acid unit is selected from natural or unnatural α, β, or γ amino acids, or a polar group such as a sugar unit (SU), polymer unit, or carboxyl unit attached to a subunit of the amino acid unit.

[0262] In some embodiments, the amino acid unit is an amino acid or a dipeptide, tripeptide, tetrapeptide, pentapeptide, hexapeptide, heptapeptide, octapeptide, nonapeptide, decapeptide, undecapeptide, or dodecapeptide, wherein one or more subunits may be modified to form a polar group, such as a sugar unit, a polymer unit, or a carboxyl unit.

[0263] In some embodiments, the subunits of the amino acid unit are selected from glycine and / or L-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and polar groups (including polymer units attached to glycine or L-amino acids). In some embodiments, the subunits of the amino acid unit are selected from glycine and / or D-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and polar groups. In some embodiments, subunits of the amino acid unit are selected from glycine and / or mixtures of L- and D-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and polar groups (including polymer units attached to glycine or a D-amino acid).

[0264] In some embodiments, the subunits of the amino acid unit are selected from glycine and / or natural L-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and at least one polar group, such as a sugar unit, polymer unit, or carboxyl unit, attached to the glycine or L-amino acid. In some embodiments, the subunits of the amino acid unit are selected from glycine and / or D-amino acids, such as arginine, glutamine, phenylalanine, tyrosine, tryptophan, lysine, alanine, histidine, serine, proline, glutamic acid, aspartic acid, threonine, cysteine, methionine, leucine, asparagine, isoleucine, and valine, and at least one polar group, such as a sugar unit, polymer unit, or carboxyl unit, attached to the glycine or D-amino acid.

[0265] In some embodiments, the subunits of the amino acid unit independently have the formula shown below in square brackets: TIFF2026503087000267.tif24128, In the formula, R 190 is 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, TIFF2026503087000268.tif59128.

[0266] In some embodiments, each subunit of the amino acid unit is independently selected from the group consisting of the following L-(naturally occurring) amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, tryptophan, and valine; and at least one polar group, e.g., a sugar unit, a polymer unit, or a carboxyl unit, attached to a natural amino acid.

[0267] In some embodiments, the subunit of the amino acid unit is not cysteine. In some embodiments, the subunit of the amino acid unit is not proline.

[0268] In some embodiments, each subunit of the amino acid unit is independently selected from the group consisting of the D-forms of these naturally occurring amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, tryptophan, and valine; and at least one polar group, e.g., a sugar unit, a polymer unit, or a carboxyl unit, attached to the glycine or L-amino acid.

[0269] In some embodiments, each subunit of the amino acid unit is independently selected from 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; and at least one polar group, e.g., a sugar unit, a polymeric unit, or a carboxyl unit, attached to one of the subunits.

[0270] Illustrative examples of alanine and its derivatives are alanine (Ala), N-alkyl-alanine, dehydro-alanine, 4-thiazolylalanine, 2-pyridylalanine, 3-pyridylalanine, 4-pyridylalanine, β-(1-naphthyl)-alanine, β-(2-naphthyl)-alanine, α-aminobutyric acid, β-chloro-alanine, β-cyano-alanine, β-cyclopentyl-alanine, β-cyclohexyl-alanine, β-iodo-alanine, β-cyclopentenyl-alanine, β-cyclopentenyl-alanine, β-cyclohexyl-alanine, β-cyclopentenyl-alanine, β-cyclopentenyl-alanine, β-cyclopentenyl-alanine, β-cyclohexyl-alanine, β-cyclopentenyl-alanine, β-cyclopentenyl-alanine, β-cyclopentenyl-alanine, β-cyclopentenyl-alanine, β-cyclohexenyl-alanine, β-cyclopent ... Examples of suitable amines include, but are not limited to, H-β-(3-benzothienyl)-Ala-OH, H-β-(2- ...

[0271] Examples of arginine and its derivatives include, but are not limited to, arginine (Arg), N-alkyl-arginine, H-Arg(Me)-OH, H-Arg(NH)-OH, H-Arg(NO)-OH, H-Arg(Ac)-OH, H-Arg(Me)-OH (asymmetric), H-Arg(Me)-OH (symmetric), 2-amino-4-(2'-hydroxyguanidino)-butyric acid (N-ω-hydroxy-nor-arginine), and homoarginine.

[0272] Examples of aspartic acid and its derivatives include, but are not limited to, aspartic acid (Asp), N-alkyl-aspartic acid, and H-Asp(OtBu)-OH.

[0273] Examples of asparagine and its derivatives include, but are not limited to, asparagine (Asn), N-alkyl-asparagine, and isoasparagine (H-Asp-NH2).

[0274] Examples of cysteine ​​(Cys) derivatives (not containing a free SH group) include, but are not limited to, H-Cys(Acm)-OH, H-Cys(Trt)-OH, H-Cys(tBu)-OH, H-Cys(Bzl)-OH, H-Cys(Et)-OH, H-Cys(SOH)-OH, H-Cys(aminoethyl)-OH, H-Cys(carbamoyl)-OH, H-Cys(phenyl)-OH, H-Cys(Boc)-OH, and H-Cys(hydroxyethyl)-OH.

[0275] Examples of histidine and its derivatives include, but are not limited to, histidine (His), N-alkyl-histidine, H-His(Boc)-OH, H-His(Bzl)-OH, H-HBs(I-Me)-OH, H-His(l-Tos)-OH, H-2,5-diiodo-His-OH, and H-His(3-Me)-OH.

[0276] Examples of glycine and its derivatives are glycine (Gly), N-alkyl-glycines, H-propargylglycine ( TIFF2026503087000269.tif18128CH) ;α - including, but not limited to, aminoglycine (protected or unprotected), β-cyclopropyl-glycine, cyclopentyl-glycine, cyclohexyl-glycine, α-allylglycine, t-butyl-glycine, neopentylglycine, and phenylglycine.

[0277] Examples of glutamic acid and its derivatives include, but are not limited to, glutamic acid (Glu), N-alkyl-glutamic acid, H-Glu(OtBu)-OH, H-γ-hydroxy-Glu-OH, H-γ-methylene-Glu-OH, H-γ-carboxy-Glu(OtBu)-OH, and pyroglutamic acid.

[0278] Examples of glutamine and its derivatives include, but are not limited to, glutamine (Gln), N-alkyl-glutamine, isoglutamine (H-Glu-NH2), H-Gln(Trt)-OH, and H-Gln(isopropyl)-OH.

[0279] Examples of phenylalanine and its derivatives include, but are not limited to, phenylalanine (Phe), N-alkyl-phenylalanine, Hp-amino-Phe-OH, Hp-amino-Phe(Z)-OH, Hp-bromo-Phe-OH, Hp-benzyl-Phe-OH, Hp-tBu-Phe-OH, Hp-carboxy-Phe(OtBu)-OH, Hp-carboxy-Phe-OH, Hp-cyano-Phe-OH, Hp-fluoro-Phe-OH, H-3,4-dichloro-Phe-OH, Hp-iodo-Phe-OH, Hp-nitro-Phe-OH, Hp-methyl-Phe-OH, H-pentafluoro-Phe-OH, Hm-fluoro-Phe-OH, H-α-Me-Phe-OH, H-4-phenyl-Phe-OH, homophenylalanine, chloro-phenylalanine, and β-homophenylalanine.

[0280] Examples of lysine and its derivatives include, but are not limited to, lysine (Lys), N-alkyl-lysine, H-Lys(Boc)-OH, H-Lys(Ac)-OH, H-Lys(formyl)-OH, H-Lys(Me)-OH, H-Lys(nicotinoyl)-OH, H-Lys(Me)-OH, H-trans-4,5-dehydro-Lys-OH, H-Lys(Aloc)-OH, HH-δ-hydroxy-Lys-OH, H-δ-hydroxy-Lys(Boc)-OH, H-Lys(acetamidoyl)-OH, and H-Lys(isopropyl)-OH. Examples of leucine and its derivatives include, but are not limited to, leucine (Leu), N-alkyl-leucine, 4,5-dehydroleucine, H-α-Me-Leu-OH, homoleucine, norleucine, and t-leucine.

[0281] Examples of methionine and its derivatives include, but are not limited to, methionine (Met), H-Met(O)-OH, and H-Met(O)2-OH.

[0282] Examples of serine and its derivatives include, but are not limited to, serine (Ser), N-alkyl-serine, H-Ser(Ac)-OH, H-Ser(tBu)-OH, H-Ser(Bzl)-OH, H-Ser(ρ-chloro-Bzl)-OH, H-β-(3,4-dihydroxyphenyl)-Ser-OH, H-β-(2-thienyl)-Ser-OH, isoserine, N-alkyl-isoserine, and 3-phenylisoserine.

[0283] Examples of tyrosine and its derivatives include, but are not limited to, tyrosine (Tyr), N-alkyl-tyrosine, H-3,5-dinitro-Tyr-OH, H-3-amino-Tyr-OH, H-3,5-dibromo-Tyr-OH, H-3,5-diiodo-Tyr-OH, H-Tyr(Me)-OH, H-Tyr(tBu)-OH, H-Tyr(Boc)-OH, H-Tyr(Bzl)-OH, H-Tyr(Et)-OH, H-3-iodo-Tyr-OH, and H-3-nitro-Tyr-OH.

[0284] Examples of threonine and its derivatives include, but are not limited to, threonine (Thr), N-alkyl-threonine, allo-threonine, H-Thr(Ac)-OH, H-Thr(tBu)-OH, and H-Thr(Bzl)-OH.

[0285] Examples of isoleucine and its derivatives include, but are not limited to, isoleucine (He), N-alkyl-isoleucine, allo-isoleucine, and norleucine.

[0286] Examples of tryptophan and its derivatives include, but are not limited to, tryptophan (Tip), N-alkyl-tryptophan, H-5-Me-Trp-OH, H-5-hydroxy-Trp-OH, H-4-Me-Trp-OH, H-α-Me-Trp-OH, H-Trp(Boc)-OH, H-Trp(formyl)-OH, and H-Trp(mesitylene-2-sulfonyl)-OH.

[0287] Examples of proline and its derivatives include, but are not limited to, proline (Pro), N-alkyl-proline, homoproline, thioproline, hydroxyproline (H-Hyp-OH), H-Hyp(tBu)-OH, H-Hyp(Bzl)-OH, H-3,4-dehydro-Pro-OH, 4-keto-proline, α-Me-Pro-OH, and H-4-fluoro-Pro-OH.

[0288] Examples of valine and its derivatives include, but are not limited to, valine (Val), N-alkyl-valine, H-α-Me-Val-OH, and norvaline.

[0289] Examples of ornithine and its derivatives include, but are not limited to, ornithine, N-alkyl-ornithine, H-Orn(Boc)-OH, H-Om(Z)-OH, H-α-difluoro-Me-Orn-OH (eflornithine), and H-Orn(Aloc)-OH.

[0290] Examples of penicillamine and its derivatives include, but are not limited to, penicillamine, H-penicillamine (Acm)-OH (H-β,β-dimethylcis(Acm)-OH), and N-alkyl-penicillamine.

[0291] Examples of β-alanine and its derivatives include, but are not limited to, β-alanine, N-alkyl-β-alanine, and dehydro-alanine.

[0292] Illustrative examples of aminoalkanoic acids and derivatives thereof include, but are not limited to, N-alkylaminoalkanoic acids, aminobutyric acid, 4-(neopentyloxysulfonyl)-aminobutyric acid, ε-aminocaproic acid, α-aminoisobutyric acid, piperidylacetic acid, 3-aminopropionic acid, 3-amino-3-(3-pyridyl)-propionic acid, and 5-aminopentanioic acid (aminovaleric acid).

[0293] Illustrative examples of aminoalkynoic acids and derivatives thereof include, but are not limited to, N-alkylaminoalkynoic acids, 6-amino-4-hexynoic acid, 6-(Boc-amino)-4-hexynoic acid.

[0294] Illustrative examples of aminoalkanedioic acids and derivatives thereof include, but are not limited to, N-alkylaminoalkanedioic acids, 2-aminohexanedioic acid, 2-aminoheptanedioic acid, 2-aminooctanedioic acid (H-Asu-OH).

[0295] Illustrative examples of aminobenzoic acids and derivatives thereof include, but are not limited to, N-alkylaminobenzoic acids, 2-aminobenzoic acid, 3-aminobenzoic acid, and 4-aminobenzoic acid.

[0296] Illustrative examples of amino-heterocyclo-alkanoic acids and derivatives thereof include, but are not limited to, N-alkylamino-heterocyclo-alkanoic acids, 4-amino-1-methyl-1H-imidazole-2-carboxylic acid, 4-amino-1-methyl-1H-pyrrole-2-carboxylic acid, 4-amino-piperidine-4-carboxylic acid (H-Pip-OH; 1-protected or not), 3-amino-3-(3-pyridyl)-propionic acid.

[0297] Illustrative examples of heterocyclo-carboxylic acids and derivatives thereof include, but are not limited to, azetidine-2-carboxylic acid, azetidine-3-carboxylic acid, piperidine-4-carboxylic acid, and thiazolidine-4-carboxylic acid.

[0298] Examples of citrulline and its derivatives include, but are not limited to, citrulline (cit), N-alkyl-citrulline, thiocitrulline, S-methyl-thiocitrulline, and homocitrulline.

[0299] Examples of statins and derivatives thereof include, but are not limited to, statins, N-alkyl-statins, cyclohexylstatins, and phenylstatins.

[0300] Illustrative examples of diaminoalkanoic acids (Dab) and derivatives thereof include, but are not limited to, N-alkyl-diamino-alkanoic acids, N,N-dialkylamino-alkanoic acids, α,γ-diaminobutyric acid (H-Dab-OH), H-Dab(Aloc)-OH, H-Dab(Boc)-OH, H-Dab(Z)-OH, α,β-diaminopropionic acid and side chain protected forms thereof.

[0301] In some embodiments, the amino acid unit may be terminated with a capping group such as a straight or branched alkyl group, or a polyethylene chain (1-30 subunits) or polymer unit.

[0302] Exemplary embodiments of amino acid units include the following, where SU is a sugar unit, poly is a polymer unit, and CU is a carboxyl unit:

[0303] In some embodiments, the amino acid unit comprises SU.

[0304] In some embodiments, the amino acid unit comprises SU-Lys-SU.

[0305] In some embodiments, the amino acid unit comprises SU-Lys-SU-tert-butyl.

[0306] In some embodiments, the amino acid unit comprises SU-Lys.

[0307] In some embodiments, the amino acid unit comprises Lys-SU.

[0308] In some embodiments, the amino acid unit comprises Lys-SU-Lys (poly).

[0309] In some embodiments, the amino acid unit comprises SU-Lys(poly)-SU.

[0310] In some embodiments, the amino acid unit comprises SU-Glu-SU.

[0311] In some embodiments, the amino acid unit comprises Lys(poly).

[0312] In some embodiments, the amino acid unit comprises Lys(poly)-Lys(poly).

[0313] In some embodiments, the amino acid unit comprises CU.

[0314] In some embodiments, the amino acid unit comprises CU-CU.

[0315] In some embodiments, an amino acid unit is present and is attached to the peptide of linker subunit L2 via a peptide bond. In some embodiments, such an amino acid unit-linker subunit L2 comprises SU-Val-Cit~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such an amino acid unit-linker subunit L2 comprises SU-Val-Ala~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such an amino acid unit-linker subunit L2 comprises SU-Val-Lys~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such an amino acid unit-linker subunit L2 comprises SU-Gly-Gly-Phe-Gly~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit.

[0316] In some embodiments, such amino acid unit-linker subunit L2 comprises Val-Lys(poly)~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such amino acid unit-linker subunit L2 comprises Val-Cit(poly)~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such amino acid unit-linker subunit L2 comprises Lys(poly)-Val-Cit~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such amino acid unit-linker subunit L2 comprises Lys(poly)-Gly-Gly-Phe-Gly~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit.

[0317] In some embodiments, such amino acid unit-linker subunit L2 comprises CU-Val-Cit~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such amino acid unit-linker subunit L2 comprises CU-Val-Lys~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such amino acid unit-linker subunit L2 comprises CU-Val-Ala~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit. In some embodiments, such amino acid unit-linker subunit L2 comprises Val-CU~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit, and CU comprises a lysine residue. In some embodiments, such amino acid unit-linker subunit L2 comprises CU-Gly-Gly-Phe-Gly~, where the wavy line indicates the bond to the remainder of the linker subunit L2 or to the Drug unit.

[0318] In some embodiments, the amino acid units are present and are attached to the linker subunit L2 by non-peptide bonds. 10 Alkylene, C2-C10 Alkenylene, C2-C 10 It is attached to the linker subunit L2 by an alkynylene or peptide linking group such as polyethylene glycol.

[0319] In some embodiments, a linker intermediate or linker is provided, wherein L2 or AA-L2 has the following structure: TIFF2026503087000270.tif187135, In the formula, the wavy line over the amino group indicates the attachment site for the Stretcher unit, and the Drug unit is attached to the benzyl alcohol.

[0320] Stretcher unit (L1) The Stretcher unit (L1) can link a targeting unit to an amino acid unit (AA) or a linker subunit L2. The Stretcher unit has a functional group that can form a bond with a functional group of a targeting unit. In some embodiments of the linker, the Stretcher unit is attached to an amino acid unit, which is attached to a linker subunit L2 (i.e., when s of AA is 1; see, e.g., Formulas (I)-(IV)). In some embodiments, the Stretcher unit is attached to a linker subunit L2 (i.e., when s of AA is 0; see, e.g., Formulas (I)-(IV)). In some embodiments, the Stretcher unit is attached to the amino acid unit-linker subunit L2 after the amino acid unit-linker subunit L2 is formed. In some embodiments, the Stretcher unit is attached to the amino acid unit-linker subunit L2-drug unit after the amino acid unit-linker subunit L2-drug unit is formed. In some embodiments, a Stretcher unit is attached to the Linker sub-unit L2-Drug unit after the Linker sub-unit L2-Drug unit is formed.

[0321] The functional group of the Stretcher unit for attachment to the targeting unit can include, for example, a maleimide, a haloacetamide, a sulfhydryl group, an NHS ester, an aldehyde, a ketone, a carbonyl, a hydrazide, a hydroxylamine, an amine, an amino, a hydrazine, a thiosemicarbazone, a hydrazine carboxyl, or an arylhydrazide.

[0322] Functional groups that may be present on the targeting unit naturally or through chemical manipulation include, but are not limited to, sulfhydryl (-SH), amino, hydroxyl, carboxy, anomeric hydroxyl groups of carbohydrates, and carboxyl groups. In one aspect, the functional groups of the targeting unit are sulfhydryl and amino. Sulfhydryl groups can be generated by reducing the intramolecular disulfide bond of the targeting unit. Alternatively, sulfhydryl groups can be generated by reacting the amino group of the lysine moiety of the targeting unit with 2-iminothiolane (Traut's reagent) or other sulfhydryl group-generating reagents.

[0323] In some embodiments, the Stretcher unit forms a bond with a sulfur atom of the targeting unit via a maleimide group of the Stretcher unit. The sulfur atom can be derived, for example, from a sulfhydryl group of the targeting unit (e.g., a thiol group of an interchain disulfide bond). Exemplary Stretcher units of this embodiment are represented by the following formulas 100 and 101: TIFF2026503087000271.tif94128, where L is the targeting unit and the wavy line indicates the attachment site for the amino acid unit or to the linker subunit L2.

[0324] In some embodiments, the stretcher unit comprises: a linker selected from TIFF2026503087000272.tif137142 is provided; In the formula, wavy line TIFF2026503087000273.tif1128 shows the binding site of the stretcher unit to the amino acid unit.

[0325] In Equations 100 and 101, 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~C 10 Alkylene-C(=O)-, C1-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-C10 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~C10 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-. 17 Any of the substituents may be substituted or unsubstituted (also referred to as unsubstituted). In some embodiments, R 17 The substituent is unsubstituted. In some embodiments, R 17 The substituent may be substituted. In some embodiments, R 17 Groups (see, for example, WO2013 / 173337), such as -(CH) x NH2, -(CH2) x NHR a , and -(CH2) x NR a 2, where x is an integer from 1 to 4, and each R a are independently selected from the group consisting of C1-C6 alkyl and C1-C6 haloalkyl, or two R a The groups together with the nitrogen to which they are attached form an azetidinyl, pyrrolidinyl, or piperidinyl group.

[0326] In some embodiments of Formula 100, R 17 is -C1-C6 alkylene-C=O-. In some embodiments, R 17 is -C1 alkylene-C(=O)-.

[0327] In some embodiments of Formula 100, R 17 -(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), -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)- (where b is 1 to 26), -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), or -C1 to C8 alkylene-NH-(C(=O))-(CH2-O-CH2) b -C1-C8 alkylene-NH- (wherein b is 1 to 26).

[0328] In another embodiment, the Stretcher unit is attached to the Targeting unit via a disulfide bond between a sulfur atom of the Stretcher unit and a sulfur atom of the Targeting unit. An exemplary Stretcher unit of this embodiment has the following formula 102: TIFF2026503087000274.tif43128, where L is a targeting unit, the wavy line indicates the binding site for the amino acid unit or linker subunit L2, and R 17 is as described above for Equations 100 and 101.

[0329] In yet another embodiment, the reactive group of the Stretcher unit contains a reactive moiety capable of forming a bond with a primary or secondary amino group of the Targeting unit. Examples of these reactive moieties include, but are not limited to, activated esters such as succinimide esters, 4-nitrophenyl esters, pentafluorophenyl esters, tetrafluorophenyl esters, anhydrides, acid chlorides, sulfonyl chlorides, isocyanates, and isothiocyanates. Exemplary Stretcher units of this embodiment are represented by Formulas 103, 104, and 105: TIFF2026503087000275.tif70128TIFF2026503087000276.tif29128, where L is a targeting unit, the wavy line indicates the binding site for the amino acid unit or linker subunit L2, and R 17 is as described above for Equations 100 and 101.

[0330] In yet another embodiment, the reactive group of the Stretcher unit contains a reactive site that is reactive with the (-CHO) group of the modified carbohydrate that may be present on the targeting unit. For example, the carbohydrate can be mildly oxidized using a reagent such as sodium periodate, and the (-CHO) unit of the resulting oxidized carbohydrate can be condensed with a Stretcher unit containing a functional group such as hydrazide, oxime, primary or secondary amine, hydrazine, thiosemicarbazone, hydrazine carboxyl, or arylhydrazide (such as those described by Kaneko, T. et al. (1991) Bioconjugate Chem. 2:133-41). Representative Stretcher units of this embodiment are represented by Formulas 106, 107, and 108: TIFF2026503087000277.tif103128, where L is a targeting unit, the wavy line indicates the binding site for the amino acid unit or linker subunit L2, and R 17 is as described above for Equations 100 and 101.

[0331] In some embodiments, it is desirable to extend the length of the stretcher unit. Thus, the stretcher unit may include additional components. An exemplary stretcher unit of this embodiment has the following formula 109: TIFF2026503087000278.tif44128, where L is a targeting unit, the wavy line indicates the binding site for the amino acid unit or linker subunit L2, and R 17 is as described above for Equations 100 and 101.

[0332] In some aspects of this embodiment, R 17 is -C1-C5 alkylene-C(=O)-. 13 is -C1-C6 alkylene, -(CH2-O-CH2) b -(wherein b is 1 to 26), -C3-C8 carbocyclo-, -arylene-, -C1-C 10 Heteroalkylene-, -C3-C8 heterocyclo-, -C1-C 10 Alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1~C 10 Alkylene-(C3-C8 carbocyclo)-, -(C3-C8 carbocyclo)-C1-C 10 Alkylene-, -C1~C 10 Alkylene-(C3-C8 heterocyclo)- or -(C3-C8 heterocyclo)-C1-C 10 In a preferred embodiment, R 13 -(CH2-O-CH2) b - where b is 1 to 26.

[0333] Targeting Unit In some embodiments, the linker is attached to the targeting unit to form a targeting unit-linker. In some embodiments, the linker is attached to the targeting unit via a stretcher unit (L1) and to the drug unit via a linker subunit L2 to form a conjugate. In some embodiments, the linker is attached to the targeting unit via a stretcher unit (L1) and to the drug unit via a linker subunit L2 to form a conjugate. In some embodiments, the targeting unit is a protein, polypeptide, or peptide. The targeting unit may be an antibody, an antigen-binding portion thereof, or a non-antibody targeting unit. A non-antibody targeting unit may also be referred to as a non-antibody scaffold.

[0334] In some embodiments, the targeting unit specifically binds to a target molecule. As used herein, "specifically binds" refers to a targeting unit (e.g., an antibody or portion thereof) described herein specifically binds to a target molecule with 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.Specific binding refers to the ability to bind to a target.Specific binding can be affected, for example, by the affinity and avidity of targeting unit and the concentration of target polypeptide.Those skilled in the art can use any suitable method, such as titrating binding substances in suitable cell binding assays, to determine the appropriate conditions under which the antibody, antibody binding moiety and non-antibody scaffold described herein selectively bind to a target.Targeting units that specifically bind to their target are not displaced by dissimilar competitors.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.

[0335] 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).

[0336] 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.

[0337] 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).

[0338] 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).

[0339] 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).

[0340] In some embodiments, the targeting unit 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.

[0341] In some embodiments, the targeting unit is a target molecule, e.g., a cancer-associated antigen, e.g., CD19, CD20, CD30, CD33, CD38, CA125, 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), HER2, 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, TUBB3, VHL, CDH1, ERBB4, FBXW7, HNF1A, JAK3, NPM1, PTPN11, R B1, SMAD4, SMARCB1, STK1, MLH1, MSH2, MSH6, PMS2, ROS1, ERCC1, 5T4(TPBG), B7-H3, CCR7, CD105, CD22, CD46, CD47, CD56, CD70, It binds to 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. 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.

[0342] In some embodiments, the targeting unit specifically binds to a target such as CD19, CD20, CD30, CD33, CD70, LIV-1, HER2, or EGFRv3.

[0343] In some embodiments, the targeting unit 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.

[0344] In some embodiments, the targeting unit is a non-antibody scaffold. In some embodiments, the targeting unit is a non-antibody protein scaffold. Such non-antibody scaffolds include, for example, affibody, affilin, anticalin, atrimer, avimer, bicyclic peptide, cysteine ​​knot, DARPin, FN3 scaffold (e.g., adnectin, centilin, pronectin, and Tn3), finomer, Kunitz domain, and O-body. (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, for example, Vazquez-Lombardi et al., Drug Discovery Today 20(10):1271 (2015) and references cited therein.) Non-antibody scaffolds can be considered to be classified into two structural categories: domain-sized constructs (in the range of 6 to 20 kDa) and constrained peptides (in the range of 2 to 4 kDa). Domain-sized non-antibody scaffolds include, but are not limited to, affibodies, affilins, anticalins, atrimers, DARPins, FN3 scaffolds (such as adnectins and centilins), 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 sentinels), finomers, 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 underlying proteins or peptides 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.

[0345] 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.

[0346] Thus, in some embodiments, a targeting unit may comprise a non-antibody scaffold.Thus, in some embodiments, a targeting unit may comprise a non-antibody scaffold protein. Those skilled in the art will appreciate that in some embodiments, the targeting unit may be, 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.

[0347] constant region In some embodiments, a targeting unit, such as an antibody or antigen-binding portion thereof, or other targeting unit, has 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).

[0348] 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.

[0349] Furthermore, targeting units comprising antibodies or antigen-binding portions thereof or non-antibody scaffolds may be part of larger molecules formed by covalent or non-covalent binding of antibodies or antigen-binding portions to one or more other proteins or peptides. Examples of such targeting units 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).

[0350] Fc domain modifications that alter effector function In some embodiments, the Fc region or Fc domain of a targeting unit, 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).

[0351] 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.

[0352] 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.

[0353] 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.

[0354] 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).

[0355] 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)).

[0356] 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.

[0357] 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.

[0358] Targeting units, such as antibodies or antigen-binding portions thereof, or non-antigenic scaffolds, with reduced effector function activity include substitutions of one or more 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 units, such as antibodies or antigen-binding portions thereof or non-antibody scaffolds, that contain such amino acid modifications and have reduced binding to FcRs can be prepared.

[0359] In some embodiments, the targeting unit, 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.

[0360] 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).

[0361] Methods for producing antibodies and antigen-binding moieties and other targeting units In various embodiments, targeting units 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 offers alternatives to cell-based manufacturing systems. A major advantage of transgenic animals is the potential for high yields from renewable resources.

[0362] Nucleic acid molecules encoding the amino acid sequence of a targeting unit, 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 ligations, 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.

[0363] 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.

[0364] 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.

[0365] Thus, expression of targeting units, 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.

[0366] 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.

[0367] 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.

[0368] 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.

[0369] 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.

[0370] 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.

[0371] 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.

[0372] 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.

[0373] 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.

[0374] 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)).

[0375] 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).

[0376] 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.

[0377] 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).

[0378] 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.

[0379] 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.

[0380] 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.

[0381] 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).

[0382] 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.

[0383] 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).

[0384] 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.

[0385] 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).

[0386] 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.

[0387] 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).

[0388] 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).

[0389] 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).

[0390] Engineered antibodies with three or more functional antigen-binding sites, including "octopus antibodies," can also be targeting units (see, e.g., U.S. Patent Application Publication No. 2006 / 0025576A1).

[0391] In some embodiments, the targeting unit 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 unit that promote the association of the desired polypeptides.

[0392] 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.

[0393] In some embodiments, the targeting unit 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 contain 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 comprises 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.

[0394] 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 activities 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 unit 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.

[0395] In some embodiments, the targeting unit is multispecific, such as an 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.

[0396] 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.

[0397] 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).

[0398] Drug Unit In some embodiments, the linker is attached to the drug unit, the targeting unit, and / or the targeting unit and drug unit (the latter also referred to as a conjugate, ADC, or antibody-drug conjugate). In some embodiments, the linker is attached to at least one drug unit via a linker subunit L2. As used herein, in the context of a conjugate, the term "drug unit" or drug refers to a cytotoxic agent (such as a chemotherapeutic agent or drug), an immunomodulator, a nucleic acid (including siRNA), a growth inhibitor, 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.

[0399] 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.

[0400] 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 in the present invention are disclosed in U.S. Patent No. 9,345,785, which is incorporated herein by reference.

[0401] 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.

[0402] Colchicine includes, but is not limited to, colchicine and CA-4.

[0403] Vinca alkaloids include, but are not limited to, vinblastine (VBL), vinorelbine (VRL), vincristine (VCR), and vindesine (VOS).

[0404] Taxanes include, but are not limited to, paclitaxel and docetaxel.

[0405] Cryptophycins include, but are not limited to, cryptophycin-1 and cryptophycin-52.

[0406] 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.

[0407] 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 by Streptomyces sp.); and 4,5-deoxy (see U.S. Pat. No. 4,371,533) (prepared by titanium trichloride / LAH reduction of maytansinol).

[0408] Hemiasterins include, but are not limited to, hemiasterin and HTL-286.

[0409] Other tubulin disrupting agents include taccalonolide A, taccalonolide B, taccalonolide AF, taccalonolide AJ, taccalonolide Al-epoxide, discodermolide, epothilone A, epothilone B, and laulimalide.

[0410] 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, a conjugate is provided in which the cytotoxic agent is a diastereomer of exatecan. Other camptothecins are disclosed in WO1996 / 021666, WO00 / 08033, US2016 / 0229862, and WO2020 / 156189.

[0411] In some embodiments, the cytotoxic agent is a duocarmcycin, including the synthetic analogs KW-2189 and CBI-TMI.

[0412] immunomodulator In some embodiments, the Drug Unit is an immunomodulator, which may be, for example, a TLR7 and / or TLR8 agonist, a STING agonist, a RIG-I agonist, or other immunomodulator.

[0413] 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, heteroarothiadiazide-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, heteroarothiadiazide-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.

[0414] 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.

[0415] In some embodiments, the TLR8 agonist may be any of the compounds described in WO2018 / 170179, WO2020 / 056198, and WO2020056194.

[0416] Other TLR7 and TLR8 agonists are described, for example, in WO2016142250, WO2016142250, WO2017046112, WO2007024612, WO2011022508, WO2011022509, WO2012045090, WO2012097173, WO2012097177, WO2 017079283, US20160008374, US20160194350, US20160289229, US Patent No. 6043238, US20180086755, WO2017216054, WO2017190669, WO2017202704, WO2017202703, WO20170071944 , US20140045849, US20140073642, WO2014056953, WO2014076221, WO2014128189, US20 140350031, WO2014023813, US20080234251, US20080306050, US20100029585, US201100 92485, US20110118235, US20120082658, US20120219615, US20140066432, US20140088085, US20140275167, and US20130251673, WO2018198091, and US20170131421.

[0417] 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.

[0418] 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.

[0419] 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.

[0420] 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.

[0421] 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.

[0422] 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.

[0423] Drug Loading A conjugate may contain one or more drug units per targeting unit. The number of drug units per targeting unit is called the drug loading. The drug loading of a conjugate is the average number of drug units (drug molecules (e.g., cytotoxic agents)) per targeting unit (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 units (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 load In 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.

[0424] The average number of drug units per targeting unit (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.

[0425] 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 unit (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, a linker is first attached to a drug unit (e.g., a cytotoxic agent, an immunomodulator, or other agent), and then a drug-linker is attached to a targeting unit (e.g., an antibody or its antigen-binding portion or a non-antibody protein scaffold). In some embodiments, a linker is first attached to a targeting unit (e.g., an antibody or its antigen-binding portion or a non-antibody protein scaffold), and then a 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 unit; 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 Unit 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).

[0426] 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 the nucleophilic group of a targeting unit (e.g., an antibody or its antigen-binding portion, or a non-antibody protein scaffold) to form a targeting unit-linker intermediate via a covalent bond, followed by reaction with a drug unit; and (2) the reaction of a bivalent linker with the 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 unit. 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.

[0427] Nucleophilic groups on targeting units 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 units, 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. Each cysteine ​​bridge thus theoretically forms two reactive thiol nucleophiles. Additional nucleophilic groups can be introduced into targeting units, 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 units (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).

[0428] Conjugates can also be generated by the reaction of an electrophilic group on a targeting unit, such as an aldehyde or ketone carbonyl group, with a nucleophilic group on a linker reagent. Useful nucleophilic groups on a linker reagent 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, for example, 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, for example, 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 agent, including non-antibody scaffolds) that can react with appropriate groups on a linker (see, e.g., Hermanson, Bioconjugate Techniques). In another embodiment, targeting units such as antibodies containing an N-terminal serine or threonine residue 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.

[0429] 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.

[0430] 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 comprises 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.

[0431] 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.

[0432] 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).

[0433] 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).

[0434] 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.

[0435] 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.

[0436] 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.

[0437] 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).

[0438] 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).

[0439] 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.

[0440] Pharmaceutical preparations 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.

[0441] 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, and the like, and combinations thereof. Additionally, 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). The pharmaceutical compositions described herein can include pharmaceutically acceptable salts of the components therein. Pharmaceutically acceptable salts include acid addition salts (formed with the free amino groups of a polypeptide) formed with inorganic acids such as hydrochloric or phosphoric acid, or organic acids such as acetic, tartaric, or mandelic acid. Salts formed with free carboxylic 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, histystadine, and prodecane. Physiologically acceptable carriers are well known in the art. An exemplary liquid carrier is a sterile aqueous solution, such as phosphate-buffered saline, containing the active ingredient (e.g., complex) and water, which 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 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 tha...

Claims

1. A linker compound comprising: (a) a Linker unit having 1 to 4 attachment sites per Drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to said amino acid unit, said polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, said polymer unit having the formula: ~R 0 -(R 3 -R 1 -[O-CH 2 -CH 2 ] n0 -R 6 -([O-CH 2 -CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 ) n3 (Ia) or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of the amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C 1 ~C 6 alkylene; Each R 3 are independently bonded, C 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)-, -C 1 ~C 12 Alkylene-NR a -C(O)-, -C 1 ~C 12 Alkylene-C(O)-NR a -C 1 ~C 12 Alkylene-, -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)-, -NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-, -C(O)-NR a -C 1 ~C 12 Alkylene-(CH(OH)) 1～8 -C 1 ~C 12 Alkylene-, -O-CH 2 -CH 2 , -OC(O)-NR a -C 1 ~C 12 Alkylene, -O-CH 2 -CH(OH)-C(O)-, -O-CH 2 -CH(OH)-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C 1 ~C 12 Alkylene-, C 1 ~C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-C 1 ~C 12 Alkylene-NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-NR a -C 1 ~C 12 Alkylene-, -NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-NR a -C 1 ~C 12 Alkylene-, -[C(O)-(CH 2 ) 1～8 -NR a ] 1～8 -, triazolyl, -C 1 ~C 12 Alkylene-triazolyl-, -N(polyhydroxyl group)-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C 1 ~C 12 alkylene and the other is C 1 ~C 12 alkylene, and each R a are independently H, C 1～6 alkyl, and wherein any of the above alkylene groups is selected from -SO 3 optionally substituted with H; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R 6 teeth, (i) In the above formula, each n 3 and n 4 are independently between 0 and 1, Each R b are independently H or C 1～6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH) 2 , or -(CH 2 ) v -OS(=O) 2 (OH), each p is independently 0 to 6; m is 1 to 4; each v is independently 1 to 6; n 2 is 1; (ii) In the above formula, Each R a are independently H or C 1～6 is alkyl, Each R b are independently H or C 1～6 is alkyl, n 6 is between 1 and 10, each p is independently 0 to 6; n 2 is 1; (iii) In the above formula, Each R a are independently H or C 1～6 is alkyl, Each R b are independently H or C 1～6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH) 2 , or -(CH 2 ) v -OS(=O) 2 (OH), each p is independently 0 to 6; q is 1 to 8, each v is independently 1 to 6; n 2 is 1; (iv) In the above formula, Each R a are independently H or C 1～6 is alkyl, Each R b are independently H or C 1～6 is alkyl, each p is independently 0 to 6; n 2 is 1; (v) -R 10 -[O-CH 2 -CH 2 ] 1～8 -R 10 - In the above formula, Each R b are independently H or C 1～6 is alkyl, Each R 10 is independent and each p is independently 1 to 6; Each R 9 are independently H, acetyl, -P(=O)(OH) 2 , or -(CH 2 ) v -OS(=O) 2 (OH), q is 1 to 8, n 2 is 1; and (vi) -N-(R 1 -X-R 2 -) 2 In the above formula, Each X is independently -NR a -C(O)- or -C(O)NR a - and n 2 is 2; Selected from: The wavy line (~) is R 0 indicating the binding site of the amino acid unit to each n 0 are independently 2 to 26; each n 1 are independently 1 to 6; n 3 is 1 to 6, The polar group.

2. A linker compound comprising: (a) a Linker unit having 1 to 4 attachment sites per Drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to said amino acid unit, said polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, said polymer unit having the formula: ~R 0 -(R 3 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -(NR 4 R 5 ) n1 ) n3 (Ia') or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of the amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C 1 ~C 6 alkylene; Each R 3 are independently -N(polyhydroxyl group)-, triazolyl, -C 1 ~C 12 alkylene-triazolyl-, and Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R a are independently H or C 1～6 is alkyl; is R 0 to R 3 indicates the binding site of Wavy line is R 1 to R 3 indicates the binding site of each p is 1 to 6; each n 0 are independently 2 to 8; each n 1 are independently 1 to 6; n 3 is 1 to 6, The polar group.

3. A linker compound comprising: (a) a Linker unit having 1 to 4 attachment sites per Drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to an amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit having the formula: ~R 0 -(R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n3 (Ia") or a stereoisomer or salt thereof, wherein: (i) R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C 1 ~C 6 alkylene; R 3 is —C(O)—; R 4 is H; R 5 is independently a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to n 0 are independently 2 to 26; n 1 is 1 to 6; n 3 is 1 to 6; (ii) R 0 is —C(O)—; R 1 , R 2 , and R 3 are each bonds; R 4 and R 5 are each independently H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to n 0 is 6; n 1 is 1 to 6; n 3 is 1; (iii) R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 6 alkylene; R 3 Ha-NR a -C(O)-C 1 ~C 12 alkylene-C(O)-, where the alkylene is -SO 3 is substituted with H; R a is H or C 1～6 is alkyl; R 4 and R 5 are each independently H, a carboxyl-containing moiety, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; The wavy line (~) is R 0 indicates the binding site of the amino acid unit to each n 0 are independently 1 to 26; n 1 is 1 to 6; n 3 is 1 to 6; or (iv) R 0 teeth and Each R 1 are independently a bond or C 1 ~C 6 alkylene; R 2 and R 3 are each bonds; R 4 and R 5 are each independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R a are independently H or C 1～6 is alkyl; Wavy line is the R 0 indicates the binding site of Wavy line (~*) is R 0 indicates the binding site of the amino acid unit to n 0 is 1 to 8; n 1 is 1 to 6; n 3 is 2, The polar group.

4. A linker compound comprising: (a) A linker unit having 1 to 4 attachment sites per Drug unit, the linker unit having the formula: or a stereoisomer or salt thereof, wherein α- represents the direct or indirect attachment site to the amino acid unit; δ- represents an attachment site to at least one of the Drug units or for a linking group attached to at least one of the Drug units; R a is H or C 1～6 is alkyl, the linker unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to said amino acid unit, said polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit.

5. A linker compound comprising: (a) a Linker unit having 1 to 4 attachment sites per Drug unit; (b) an amino acid unit having 1 to 12 amino acid subunits; and (c) at least one polar group attached to the amino acid unit, the polar group comprising a polymer unit, optionally a sugar unit, and optionally a carboxyl unit, the polymer unit comprising: (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, 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, or a stereoisomer thereof; or (iii) any combination thereof The polar group comprises:

6. At least one polar group attached to said amino acid unit has the formula: ~R 0 -(R 3 -R 1 -[O-CH 2 -CH 2 ] n0 -R 6 -([O-CH 2 -CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 ) n3 (Ia), ~R 0 -(R 3 -R 1 -[O-CH 2 -CH(OH)-CH 2 ] n0 -R 6 -[O-CH 2 -CH(OH)-CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n3 (Ib), or or a stereoisomer or salt thereof, wherein: R 0 is a functional group for attachment of the amino acid unit to a subunit; Each R 1 and R 2 are independently a bond or C 1 ~C 6 alkylene; Each R 3 are independently bonded, C 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)-, -C 1 ~C 12 Alkylene-NR a -C(O)-, -C 1 ~C 12 Alkylene-C(O)-NR a -C 1 ~C 12 Alkylene-, -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)-, -NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-, -C(O)-NR a -C 1 ~C 12 Alkylene-(CH(OH)) 1～8 -C 1 ~C 12 Alkylene-, -O-CH 2 -CH 2 , -OC(O)-NR a -C 1 ~C 12 Alkylene, -O-CH 2 -CH(OH)-C(O)-, -O-CH 2 -CH(OH)-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C 1 ~C 12 Alkylene-, C 1 ~C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-C 1 ~C 12 Alkylene-NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-NR a -C 1 ~C 12 Alkylene-, -NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-NR a -C 1 ~C 12 Alkylene-, -[C(O)-(CH 2 ) 1～8 -NR a ] 1～8 -, triazolyl, -C 1 ~C 12 Alkylene-triazolyl-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C 1 ~C 12 alkylene and the other is C 1 ~C 12 alkylene, and each R a are independently H, C 1～6 alkyl, and wherein any of the above alkylene groups is selected from -SO 3 optionally substituted with H; Each R 4 and R 5 are independently H, a polyhydroxyl group, a carboxyl-containing moiety, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a polyhydroxyl-ether group, a substituted polyhydroxyl-ether group, or a chelating agent, wherein the optional substituents are selected from sulfate, phosphate, alkyl sulfate, and alkyl phosphate, and wherein R 4 and R 5 At least one of them is not H; Each R 6 are independently a bond, or: (i) In the above formula, each n 3 and n 4 are independently between 0 and 1, Each R b are independently H or C 1～6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH) 2 , or -(CH 2 ) v -OS(=O) 2 (OH), each p is independently 0 to 6; m is 1 to 4; each v is independently 1 to 6; n 2 is 1; (ii) In the above formula, Each R a are independently H or C 1～6 is alkyl, Each R b are independently H or C 1～6 is alkyl, n 6 is between 1 and 10, each p is independently 0 to 6; n 2 is 1; (iii) In the above formula, Each R a are independently H or C 1～6 is alkyl, Each R b are independently H or C 1～6 is alkyl, Each R 9 are independently H, acetyl, -P(=O)(OH) 2 , or -(CH 2 ) v -OS(=O) 2 (OH), each p is independently 0 to 6; q is 1 to 8, each v is independently 1 to 6; n 2 is 1; (iv) In the above formula, Each R a are independently H or C 1～6 is alkyl, Each R b are independently H or C 1～6 is alkyl, each p is independently 0 to 6; n 2 is 1; (v) -R 10 -[O-CH 2 -CH 2 ] 1～8 -R 10 - In the above formula, Each R b are independently H or C 1～6 is alkyl, Each R 10 is independent and each p is independently 1 to 6; q is 1 to 8; and (vi) -N-(R 1 -X-R 2 -[O-CH 2 -CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) 2 In the above formula, Each X is independently -NR a -C(O)- or -C(O)NR a - and n 2 is 2; Selected from: The wavy line (~) is R 0 indicating the binding site of the amino acid unit to each n 0 are independently 2 to 26; n 1 is 0 to 6, and n 1 If is 0, R 3 is -OH or -C(O)OR b where R b are independently H or C 1～6 is alkyl; n 3 is 1 to 6, 6. The linker compound of claim 4 or 5.

7. Each R 3 are independently a bond, -C(O)-, or -NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-, -C(O)-NR a -C 1 ~C 12 Alkylene-(CH(OH)) 1～8 -C 1 ~C 12 Alkylene-, -O-CH 2 -CH(OH)-C(O)-, -O-CH 2 -CH(OH)-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-, -CH(OH)-C 1 ~C 12 Alkylene-, C 1 ~C 12 Alkylene -CH(OH)-, -CH(OH)-C(O)-, -CH(OH)-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-C 1 ~C 12 Alkylene-NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-NR a -C 1 ~C 12 Alkylene-, -NR a -C(O)-C 1 ~C 12 Alkylene-C(O)-NR a -C 1 ~C 12 Alkylene-, -CH(OH)-NR a -C 1 ~C 12 Alkylene-, -[C(O)-(CH 2 ) 1～8 -NR a ] 1～8 -, triazolyl, and -C 1 ~C 12 alkylene-triazolyl-, -N(polyhydroxyl group)-, and each R a are independently H, C 1～6 alkyl, wherein any of the above alkylene groups is selected from -SO 3 7. The linker compound of any one of claims 1 or 4 to 6, optionally substituted with H.

8. said linker unit or a stereoisomer or salt thereof, During the ceremony, α- represents the direct or indirect attachment site to said amino acid unit; δ- represents a site of attachment to at least one of the Drug units or to a linking group attached to at least one of the Drug units; R a is H or C 1～6 is alkyl, The linker compound of any one of claims 1 to 3 or 5 to 7.

9. The at least one polar group has the formula: L3-N(CH 2 -(CH(X 1 R)) k -X 2 (X 3 )) 2 (X) or a stereoisomer or salt thereof, During the ceremony, each X 1 is independently selected from NH or O; each R is independently selected from hydrogen, acetyl, monosaccharides, disaccharides, and polysaccharides; each X 2 is independently CH 2 and C(O); each X 3 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 8.

10. The at least one polar group 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 8.

11. below: (a) ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -NR 4 R 5 (XX) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 3 alkylene; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5 Both of these cannot be H; n0 is 2 to 26; (b) ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -NR 4 R 5 (XXI) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 3 alkylene; R 4 and R 5 is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), and R 4 and R 5 the other is a polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, where R 4 and R 5 Both of these cannot be H; n0 is 2 to 26; (c) ~R 0 -[-R 6 -[R 9 -[O-CH 2 -CH 2 -] n0 -R 9 ] n1 -R 7 -NR 4 R 5 ] n7 (XXII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 6 and R 7 are each independently a bond, 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 alkylene-NH-; R 4 and R 5 is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a chelating agent, and —C(O)—R, where R is a saccharide unit of formula (XII) or (XIII); and R 4 and R 5 the other is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), and polyethylene glycol optionally having 1 to 24 ethylene glycol subunits, wherein R 4 and R 5 Both of these cannot be H; Each R 9 are independently a bond, -C(O)-, -NH-, or -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 selected from alkylene-, -NH(CO)NH-, and triazole; n0 is 2 to 26; n1 is 1 to 4; n7 is 1 to 4; (d) ~R 0 -R 1 -[-C(R α )H-C(O)-N(R N )-] n0 -R 2 -NR 4 R 5 (XXIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 is a bond, C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 , -[CH 2 -CH 2 -O] n0 -C 1 ~C 3 Alkylene- or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 -C(O)-; R 2 is C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 , -[CH 2 -CH 2 -O] n0 -C 1 ~C 3 Alkylene- or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 -C(O)-; Each R α are independently H or -R 2 -NR 4 R 5 and Each R N are independently H, C 1 ~C 6 Alkyl, or -R 2 -NR 4 R 5 and R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5 Both of these cannot be H; each n0 is independently 2 to 26; (e) ~R 0 -R 1 -[-C(R α )HC(O)-N(R N )-] n0 -R 2 -WHAT 2 R 6 (XXIV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 is a bond, C 1 ~C 3 Alkylene, or -C 1 ~C 3 Alkylene [O-CH 2 -CH 2 -] n0 and R 2 is C 1 ~C 3 Alkylene or -C 1 ~C 3 Alkylene [O-CH 2 -CH 2 -] n0 and Each R α are independently H or -R 2 -NR 4 R 5 and Each R N are independently H, C 1 ~C 6 Alkyl, or -R 2 -NR 4 R 5 and R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5 Both of these cannot be H; R 6 is H or C 1 ~C 4 is alkyl; Each n0 is independently 2 to 26, However, at least one R α or R N Ha-R 2 -NR 4 R 5 is; or (f) ~R 0 -R 1 -[C(R α )H-C(O)-N(R N )-] n0 -R 2 -N-(R 3 -NR 4 R 5 ) 2 (XXV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently bonded, C 1 ~C 3 Alkylene, or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 and Each R α are independently H or -R 2 -NR 4 R 5 and Each R N are independently H or C 1 ~C 6 is alkyl; Each R 3 is independently C 1 ~C 6 alkylene; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5 Both of these cannot be H; each n0 is independently 2 to 26; 11. The linker compound of any one of claims 4 to 10, comprising a polar group having a formula selected from:

12. R 4 and R 5 are each independently selected from H and a polyhydroxyl group, where R 4 and R 5 12. The linker compound of claim 1, wherein at least one of: is not H.

13. 13. The linker compound of claim 11 or 12, wherein the polyhydroxyl group is a linear monosaccharide optionally selected from a C6 or C5 sugar, a sugar acid, or an amino sugar.

14. 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; 14. The linker compound of claim 13.

15. below: or a stereoisomer or salt thereof, wherein each R is independently H or alkyl; and each R 39 is independently selected from H, a linear monosaccharide, and polyethylene glycol, which may have 1 to 24 ethylene glycol subunits; each n is independently 1 to 12; and the wavy line is the bond to the amino acid unit. A linker compound according to any one of claims 1 to 14.

16. R 4 and R 5 12. The linker compound of any one of claims 1 to 2 or 4 to 11, wherein one of said monosaccharides is a linear monosaccharide and the other is a cyclic monosaccharide.

17. -(NR 4 R 5 ) but the following: or a stereoisomer or salt thereof; In the formula, R 11 is a cyclic monosaccharide, 17. The linker compound of claim 16.

18. below: or a stereoisomer or salt thereof, In the formula, R 41 is a cyclic monosaccharide; the wavy line indicates the bond to the amino acid unit.

17. The linker compound of claim 16.

19. R 4 and R 5 12. The linker compound of any one of claims 1-2 or 4-11, wherein is independently a polyhydroxyl selected from cyclic monosaccharides, disaccharides, and polysaccharides.

20. -(NR 4 R 5 ) but the following: or a stereoisomer or salt thereof; In the formula, each R 12 is selected from H and a monosaccharide, a disaccharide, or a polysaccharide; R 5 is selected from cyclic monosaccharides, disaccharides, or polysaccharides; 20. The linker compound of claim 19.

21. 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 indicates the bond to the amino acid unit.

20. The linker compound of claim 19.

22. R 4 and R 5 are independently selected from linear monosaccharides and substituted linear monosaccharides, wherein the substituted linear monosaccharides are substituted with monosaccharides, disaccharides, or polysaccharides.

23. -(NR 4 R 5 ) but the following: or a stereoisomer or salt thereof; In the formula, R 13 is a linear monosaccharide; each R 14 is selected from monosaccharides, disaccharides, and polysaccharides; 23. The linker compound of claim 22.

24. 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 the bond to the amino acid unit, 23. The linker compound of claim 22.

25. R 4 and R 5 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.

26. -(NR 4 R 5 ) but the following: or a stereoisomer or salt thereof; In the formula, each R 15 are independently selected from linear monosaccharides and substituted linear monosaccharides; each R 16 are independently selected from hydroxyl, carboxyl, ester, and amide; 26. The linker compound of claim 25.

27. 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 is the bond to the amino acid unit.

26. The linker compound of claim 25.

28. R 4 and R 5 is a -C(O)-polyhydroxyl group or a substituted -C(O)-polyhydroxyl group, and R 4 and R 5 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.

29. -(NR 4 R 5 ) but the following:

29. The linker compound of claim 28, selected from:

30. below: or a stereoisomer or salt thereof, wherein the wavy line is the bond to the amino acid unit.

29. The linker compound of claim 28.

31. -(NR 4 R 5 ) but the following: or a stereoisomer or salt thereof; In the formula, R 18 OH, CH 2 OH, COOH, or -C substituted with hydroxyl or carboxyl 1 ~C 6 selected from alkyl, 12. The linker compound of any one of claims 1-2 or 4-11.

32. below: or a stereoisomer or salt thereof, In the formula, R 48 OH, CH 2 OH, COOH, or -C substituted with hydroxyl or carboxyl 1 ~C 6 alkyl; the wavy line is the bond to the amino acid unit; 12. The linker compound of any one of claims 1-2 or 4-11.

33. -(NR 4 R 5 ) but the following:

12. The linker compound of any one of claims 1-2 or 4-11, selected from:

34. below: or a stereoisomer or salt thereof, wherein the wavy line is the bond to the amino acid unit.

12. The linker compound of any one of claims 1-2 or 4-11.

35. R 4 and R 5 are independently selected from H and a chelating agent, with the proviso that R 4 and R 5 12. The linker compound of any one of claims 1-2 or 4-11, wherein both of are not H.

36. The chelator is linked by an alkylene, arylene, carbocyclyl, heteroarylene, or heterocarbocyclyl to -NR 4 R 5 36. The linker compound of claim 35, wherein the linker compound is optionally attached to a nitrogen of

37. 38. The linker compound of claim 37, 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.

38. below: or a stereoisomer or salt thereof, wherein the wavy line is the bond to the amino acid unit.

38. The linker compound of claim 37.

39. R 4 and R 5 12. The linker compound of any one of claims 1-2 or 4-11, wherein: are independently selected from H, a polyhydroxyl ether group, and a substituted polyhydroxyl ether group.

40. -(NR 4 R 5 ) but the following:

40. The linker compound of claim 39, selected from:

41. below: (a) ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -R 3 (XXX) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 3 an alkylene group; R 3 may be substituted C 3 ~C 10 or R is selected from a carbocycle, a thiourea, an optionally substituted thiourea, a urea, an optionally substituted urea, a sulfamide, an alkylsulfamide, an acylsulfamide, an optionally substituted alkylsulfamide, an optionally substituted acylsulfamide, a sulfonamide, an optionally substituted sulfonamide, a guanidine including an alkylguanidine and an arylguanidine, a phosphoramide, or an optionally substituted phosphoramide; or R 3 represents azido, alkynyl, substituted alkynyl, -NH-C(O)-alkynyl, -NH-C(O)-alkynyl-R 5 , cyclooctyne; -NH-cyclooctyne, -NH-C(O)-cyclooctyne, or -NH-(cyclooctyne) 2 where R 5 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; n0 is 2 to 26; (b) ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -NH-C(O)-R 3 (XXXI) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 3 an alkylene group; R 3 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 4 having R 4 are azide, alkynyl, and alkynyl-R 5 , cyclooctyne, or cyclooctyne-R 5 where R 5 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; n0 is 2 to 26; (c) ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -C(O)NH-R 3 (XXXII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 3 an alkylene group; R 3 is a branched polyethylene glycol chain, each branch independently having 1 to 26 ethylene glycol subunits and each branch having R at its end 4 having R 4 are azide, alkynyl, and alkynyl-R 5 , cyclooctyne, or cyclooctyne-R 5 where R 5 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; n0 is 2 to 26; (d) ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -C(O)NR 3 -R 2 -NR 4 R 5 (XXXIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 3 is H or R 2 -NR 4 R 5 and R 1 and R 2 are each independently a bond or C 1 ~C 3 an alkylene group; R 4 and R 5 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, where R 4 and R 5 Both of these cannot be H; n0 is 2 to 26; (e) ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -NR 6 -R 3 (XXXIV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 3 an alkylene group; R 3 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 4 having R 6 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 4 are azide, alkynyl, and alkynyl-R 5 , cyclooctyne, or cyclooctyne-R 5 where R 5 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; n0 is 2 to 26; (f) ~R 0 -(R 1 -[CH 2 -CH(OR 3 )-CH 2 -O] n0 -R 6 ) n2 (XXXV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; Each R 1 are independently a bond, -O- or C 1 ~C 3 an alkylene group; Each R 3 are independently H, -[CH 2 -CH(OH)-CH 2 -O] n0 -R 6 , -C(O)-NR 4 R 5 or -C(O)N(R N )-C 1 ~C 6 Alkylene-NR 4 R 5 and R N is H or C 1 ~C 4 is alkyl; R 4 and R 5 are each independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, where R 4 and R 5 Both of these cannot be H; Each R 6 are independently H, C 1 ~C 6 Alkylene-C(OH)H-NR 7 R 8 , C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkylene-NR 7 R 8 , -C(O)-NR 4 R 5 , -C(O)N(R N )-C 1 ~C 6 Alkylene-NR 4 R 5 , C 1 ~C 6 Alkylene-C(O)NR 4 R 5 or C 1 ~C 6 Alkylene-CO 2 R 9 and Each R 9 are independently H or C 1 ~C 6 is alkyl; R 7 and R 8 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; each n0 is independently 1 to 26; n2 is 1 or 2; (g) ~R 0 -R 1 -[[CH 2 -CH 2 -O] n0 -R 2 -[CH 2 -[CH(OH)] n3 -CH 2 -O] n1 ] n2 -R 3 -NR 4 -R 5 (XXXVI) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 , R 2 and R 3 are each independently a bond or C 1 ~C 3 an alkylene group; R 4 and R 5 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, where R 4 and R 5 Both of these cannot be H; each n0 independently ranges from 0 to 26 and each n1 independently ranges from 0 to 26, provided that at least one of n0 or n1 ranges from 2 to 26; n2 is 1 to 5; each n3 is independently 1 or 2; (h) ~R 0 -(R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -N(R N )-CO 2 -[CH 2 -CH(OR 3 )-CH 2 -O] n1 -R 6 ) n5 (XXXVII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 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 4 and R 5 are each independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, where R 4 and R 5 Both of these cannot be H; Each R 3 are independently H, -[CH 2 -CH(OH)-CH 2 -O] n0 -R 6 or -C(O)N(R N )-C 1 ~C 6 Alkylene-NR 4 R 5 and Each R 6 are independently H, C 1 ~C 6 Alkylene-C(OH)H-NR 7 R 8 , C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkylene-NR 7 R 8 , -C(O)N(R N )-C 1 ~C 6 Alkylene-NR 4 R 5 , C 1 ~C 6 Alkylene-C(O)NR 4 R 5 or C 1 ~C 6 Alkylene-CO 2 R 9 and Each R 9 are independently H or C 1 ~C 6 is alkyl; R 7 and R 8 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; n0 is 2 to 26; n1 is 1 to 26; n5 is 1 or 2; (i) ~R 0 -(R 1 -[N(R N )-C(O)-[O-CH 2 -CH(OH)-CH 2 ] n0 ] n1 -R 2 -NR 4 R 5 ) n5 (XXXVIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 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 4 and R 5 are each independently selected from H, a polyhydroxyl group, or a substituted polyhydroxyl group, where R 4 and R 5 Both of these cannot be H; n0 is 2 to 26; n1 is 2 to 4; n5 is 1, 2, or 3; (j) ~R 0 -(R 1 -[C(R α )H-C(O)-N(R N )] n0 -R 2 -[CH 2 -CH 2 -O] n0 -NR 4 R 5 ) n5 (XXXIX) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond, C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 , -[CH 2 -CH 2 -O] n0 -C 1 ~C 3 Alkylene- or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 -C(O)-; Each R α are independently H or -R 2 -NR 4 R 5 and Each R N are independently H, C 1 ~C 6 Alkyl, or -R 2 -NR 4 R 5 and R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, and -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5 Both of these cannot be H; each n0 is independently 0 to 26, provided that at least one n0 is 2 to 26; n5 is 1 or 2; or (k) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 , R 2 and R 3 are each independently a bond, C 1 ~C 3 Alkylene, -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 , -[CH 2 -CH 2 -O] n0 -C 1 ~C 3 Alkylene- or -C 1 ~C 3 Alkylene-[O-CH 2 -CH 2 -] n0 -C(O)-; Each R α are independently H or -R 2 -NR 4 R 5 and Each R N are independently H, C 1 ~C 6 Alkyl, or -R 2 -NR 4 R 5 and R 4 and R 5 are each independently selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a -C(O)-polyhydroxyl group, a substituted -C(O)-polyhydroxyl group, a chelating agent, or -C(O)-R, where R is a saccharide unit of formula (XII) or (XIII), 4 and R 5 Both of these cannot be H; R 6 is H or C 1 ~C 6 is alkyl; each n0 is independently 0 to 26, provided that at least one n0 is 2 to 26; each n1 independently is 0 to 26, provided that at least one n1 is 2 to 26; 41. The linker compound of any one of claims 1 to 40, comprising a polar group having a formula selected from:

42. below: ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -NH-C(O)-R 3 (XXXI)、 ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -C(O)NH-R 3 (XXXII), and ~R 0 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -N-(R 6 -R 3 ) 2 (XXXIII); or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of said amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 3 an alkylene group; R 3 is a branched polyethylene glycol chain, each branch having 1 to 26 ethylene glycol subunits and each branch having R 4 having R 6 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 4 are azide, alkynyl, and alkynyl-R 5 , cyclooctyne, or cyclooctyne-R 5 where R 5 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 0 indicating the binding site of the amino acid unit to n0 is 2 to 26.

42. The linker compound of any one of claims 1 to 41.

43. 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 is the bond to the amino acid unit, 43. The linker compound of claim 41 or 42.

44. formula: ~R 0 -(R 3 -R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 (XL) or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of said amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 6 alkylene; Each R 3 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)-, and -C(O)NR 7 R 8 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 7 and R 8 One of the two is H or C 1 ~C 12 alkylene and the other is C 1 ~C 12 alkylene, C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom. 44 R 45 is bound to; R 4 and R 5 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, and wherein R 4 and R 5 Both of these cannot be H; n0 is 2 to 26; n 1 is 1 to 6; n2 is 1 to 6, 44. The linker compound of any one of claims 1 to 43.

45. formula: ~R 0 -(R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 (XLI) or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of said amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 6 alkylene; Each R 3 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 7 R 8 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 7 and R 8 One of the two is H or C 1 ~C 12 alkylene and the other is C 1 ~C 12 alkylene, C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom. 44 R 45 is bound to; R 4 and R 5 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, and wherein R 4 and R 5 Both of these cannot be H; n0 is 2 to 26; n 1 is 1 to 6; n2 is 1 to 6, 45. The linker compound of any one of claims 1 to 44.

46. formula: ~R 0 -(R 1 -[O-CH 2 -CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 (XLII) or a stereoisomer or salt thereof, During the ceremony, R 0 is a functional group for attachment of said amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 3 alkylene; Each R 3 are independently bonded, 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 7 R 8 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 7 and R 8 One of the two is H or C 1 ~C 6 alkylene and the other is C 1 ~C 12 alkylene, C 1 ~C 2 One of the alkylenes is NR at the nitrogen atom. 44 R 45 is bound to; R 4 and R 5 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, and wherein R 4 and R 5 Both of them cannot be H; n0 is 2 to 16; n1 is 1 to 4; n2 is 1 to 4, 46. ​​The linker compound of any one of claims 1 to 45.

47. R 0 is derived from a functional group of a precursor compound to said polymer unit, 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.

48. R 0 but has the following structure: or a stereoisomer thereof, In the formula, R is H, C 1 ~C 6 alkyl or polyhydroxyl group, n is 0-12, (*) indicates the R 0 The binding sites of each is the R to the remainder of the polymer unit 0 indicates the binding site of 48. The linker compound of any one of claims 1-2 or 4-47.

49. R 0 but has the following structure: or a stereoisomer thereof, In the formula, R is H, C 1 ~C 6 alkyl or polyhydroxyl group, n is 0-12, (*) indicates the R 0 The binding sites of each is the R to the remainder of the polymer unit 0 indicates the binding site of 49. The compound of claim 48.

50. R 3 If present, -R 3 -(NR 4 R 5 ) n1 but has the following structure: or a stereoisomer thereof, In the formula, each R a and R b are independently H or C 1～6 is alkyl, and X 4 SO 3 H, p is 0 to 8, is the R to the remainder of the polymer unit 3 indicates the binding site of 31. The linker compound of any one of claims 1-2 or 6-30.

51. R 3 If present, -R 3 -(NR 4 R 5 ) n1 but has the following structure: or a stereoisomer thereof, During the ceremony, is the R to the remainder of the polymer unit 3 indicates the binding site of 51. The linker compound of claim 50.

52. At least one -NR 4 R 5 When present, the following structure: or a stereoisomer thereof, During the ceremony, -NR to the remainder of the polymer unit 4 R 5 indicates the binding site of 52. The linker compound of any one of claims 1 to 51.

53. Prior to attachment to the linker unit, the structure: or a stereoisomer thereof, During the ceremony, (*) indicates the binding site to the amino acid unit; Each R, R a and R b are independently H or C 1 ~C 6 is alkyl; R' is H, C 1 ~C 6 Alkyl, -N(R 4 )(R 5 ) 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 4 and R 5 is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a chelating agent, or —C(O)—R, where R is a saccharide unit of formula (XII) or (XIII); and R 4 and R 5 the other is selected from H, a polyhydroxyl group, a substituted polyhydroxyl group, a —C(O)-polyhydroxyl group, a substituted —C(O)-polyhydroxyl group, a chelating agent, or —C(O)—R, where R is a saccharide unit of formula (XII) or (XIII), and polyethylene glycol, which may have 1 to 24 ethylene glycol subunits; or —NR 4 R 5 Let's go together 3 ~C 8 form a heterocycle, R 4 and R 5 Both of them cannot be H.

53. The linker compound of any one of claims 1 to 52.

54. below: (a) ~R 0 -(R 3 -R 1 -[O-CH 2 -CH 2 ] n0 -R 6 -[O-CH 2 -CH 2 ] n0 -R 2 -R 3 -(NR 4 R 5 ) n1 ) n2 (XLIII) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 and R 2 are each independently a bond or C 1 ~C 6 alkylene; Each R 3 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)-, and -C(O)NR 7 R 8 where R 7 and R 8 One of the two is H or C 1 ~C 12 alkylene and the other is C 1 ~C 12 alkylene; R 4 and R 5 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, and wherein R 4 and R 5 Both of these cannot be H; Each R 6 are independently, -NR a -, -NR a -C 1 ~C 6 Alkylene-NR a -, -NR a -C(O)-NR a -S(O) 2 -NR a -, or -NR a -C(O)-C 1～6 alkylene-; Each R a are independently H, C 1 ~C 6 alkyl, or polyhydroxyl groups; each n0 is independently 2 to 26; n1 is 1 to 6; n2 is 1 to 6; (b) ~R 0 -(R 1 -[O-CH 2 -CH 2 ] n3 -R 2 -X 1 -R 5 -X 2 -R 3 -[O-CH 2 -CH 2 ] n3 -R 4 -[X 3 -R 6 ] n4 -R 7 ) n5 (XLIV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 , R 2 , R 3 and R 4 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 5 and R 6 each independently represents a divalent polyhydroxyl group; R 7 is H, OH or C 1 ~C 6 is alkyl; each n3 is independently 0 to 26, provided that at least one n3 is 2 to 26; n4 is 0 to 10; n5 is 1 or 2; or (c) ~R 0 -R 1 -[O-CH 2 -CH 2 ] n3 -R 2 -N-(R 3 -X 1 -R 4 -[O-CH 2 -CH 2 ] n3 -(NR 4 R 5 )) 2 (XLV) or a stereoisomer or salt thereof, wherein R 0 is a functional group for attachment of an amino acid unit to a subunit; R 1 , R 3 and R 4 each independently optionally bonded or substituted C 1 ~C 6 alkylene; Each R 2 are independently bonded, 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 4 and R 5 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, and wherein R 4 and R 5 Both of these cannot be H; each n3 is independently 2 to 26; 54. The linker compound of any one of claims 1 to 53, comprising a polar group having a formula selected from:

55. Prior to attachment to the amino acid unit, the structure: and a polar group having one of During the ceremony, (*) indicates the binding site for the amino acid unit; Each R a are independently H, alkyl, or polyhydroxyl groups; R 4 and R 5 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, and wherein R 4 and R 5 Both of these cannot be H; each n is independently 1 to 12; 55. A linker compound according to any one of claims 1 to 54.

56. 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 a bond to an amino acid unit, 56. The linker compound of any one of claims 1 to 55.

57. A polar group having a formula selected from the formula: 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 a bond to an amino acid unit, 57. A linker compound according to any one of claims 1 to 56.

58. A polar group having a formula selected from the following, 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 a bond to an amino acid unit, 58. The linker compound of any one of claims 1 to 57.

59. Y is R 76 59. The linker compound of claim 58, wherein:

60. Y is 59. The linker compound of claim 58, wherein:

61. Each R a and R b 59. The linker compound of claim 58, wherein:

62. R a and R b 59. The linker compound of claim 58, wherein, together with the carbon to which they are attached, form an oxo group.

63. 59. The linker compound of any one of claims 56 to 58, wherein q is 10 to 20.

64. 59. The linker compound of any one of claims 56 to 58, wherein q is 12.

65. The polar group has the following structure prior to attachment to the amino acid unit: or a stereoisomer thereof, In the formula, R a is H or C 1～6 alkyl, and n is 1 to 20; 65. The linker compound of any one of claims 1 to 64.

66. The polar group has the following structure prior to attachment to the amino acid unit: or a stereoisomer thereof, In the formula, R a is H or C 1～6 alkyl, and n is 1 to 20; 66. The linker compound of any one of claims 1 to 65.

67. The polar group has the following structure prior to attachment to the amino acid unit: or a stereoisomer thereof, In the formula, R a is H or C 1～6 alkyl, and n is 1 to 20; 67. The linker compound of any one of claims 1 to 66.

68. below: or a stereoisomer or salt thereof, wherein each Z is attached at *; More independently selected, each is the bond to the amino acid unit, 68. The linker compound of any one of claims 1 to 67.

69. The polar group is: or a stereoisomer thereof, During the ceremony, each indicates a bond to said amino acid unit, 68. The linker compound of any one of claims 1 to 67.

70. The polar group has the following 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)-, where * is the bond to the amino acid unit or to the remainder of the 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; (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)-, where * is the bond to the amino acid unit or to the remainder of the 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, 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; 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)-, where * is the bond to the amino acid unit 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 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; 70. The linker compound of any one of claims 1 to 69.

71. 71. The linker compound of any one of claims 1 to 70, comprising a polar group comprising said polymer unit and a saccharide unit.

72. 72. The linker compound of any one of claims 1-71, comprising a polar group comprising at least two polymer units.

73. 72. The linker compound of any one of claims 1 to 71, comprising a polar group comprising said polymer unit and a carboxyl unit.

74. 74. The linker compound of any one of claims 1-73, comprising at least two polar groups.

75. 75. The linker compound of any one of claims 1 to 74, comprising a polar group comprising said polymer unit, said sugar unit, and said carboxyl unit.

76. 76. The linker compound of any one of claims 1-75, comprising a polar group comprising at least two polymer units, at least one sugar unit, and at least one carboxyl unit.

77. 77. The linker compound of any one of claims 1 to 76, wherein said amino acid unit comprises at least two amino acid subunits.

78. 78. The linker compound of any one of claims 1 to 77, wherein if present, each comprises two of said polar groups attached to said amino acid unit.

79. 79. The linker compound of any one of claims 1 to 78, wherein said linker unit is attached to a side chain of a subunit of said amino acid unit.

80. 80. The linker compound of any one of claims 1-79, wherein said amino acid unit is attached to said linker unit by a non-peptide linking group.

81. 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 81. The linker compound of claim 80, which is selected from alkynylene, or optionally substituted polyethylene glycol.

82. The following structure: or a stereoisomer thereof, wherein said polar group is attached to an amino acid subunit of said Amino Acid unit, and an H of the hydroxyl or amino group of a para-aminobenzyl group, or an H of the hydroxyl of a glycine residue of GGFG peptide, is optionally replaced with a bond to at least one of the Drug units or to a linking group attached to at least one of the Drug units, and wherein a wavy line on the amino group indicates an attachment site for a Stretcher unit or an Amino Acid unit, or indicates an H prior to attachment.

83. below: and a formula selected from wherein the square brackets represent the amino acid unit, each aa is any subunit of the amino acid unit, L2 is the linker unit, and each wavy line (~) represents a binding site for a stretcher unit; 1 (poly) is a polymer unit attached to an amino acid subunit of the amino acid unit, SU is a saccharide unit attached to a subunit of the amino acid unit or to the linker unit, and CU is a carboxyl unit attached to a subunit of the amino acid unit or to the linker unit; double wavy line represents a binding site for at least one of said Drug units, wherein aa and aa 1 are independently selected from α, β and γ amino acids and derivatives thereof; 83. The linker compound of any one of claims 1 to 82.

84. below: and a formula selected from where the square brackets represent the amino acid unit, each aa is an amino acid subunit of the amino acid unit, L2 is a linker subunit attached to the side chain of aa, and the wavy line (~) represents the attachment site for the Stretcher unit; 1 (Poly) is a polymer unit attached to aa, SU is a sugar unit attached to aa, CU is a carboxyl unit attached to aa, and the double wavy line represents a binding site for at least one of said Drug units; 1 are independently selected from α, β and γ amino acids and derivatives thereof; 83. The linker compound of any one of claims 1 to 82.

85. 83. The linker compound of any one of claims 1-82, wherein at least two polymer units are attached to said amino acid unit.

86. below: and a formula selected from In the formula, the square brackets represent the amino acid unit, aa is any subunit of the amino acid unit, L2 is the linker unit, and the wavy line (~) represents the attachment site for the stretcher unit; 1 (poly) and aa 2 (poly) is a polymer unit attached to an aa or other polymer unit; each SU is a sugar unit attached to an aa or other sugar unit, and each CU is a carboxyl unit attached to an aa or other carboxyl unit, and the double wavy line represents a binding site for at least one of said Drug units; 1 and aa 2 are independently selected from α, β and γ amino acids and derivatives thereof; 83. The linker compound of any one of claims 1 to 82.

87. below: and a formula selected from where the square brackets represent the amino acid unit, aa is an amino acid subunit of the amino acid unit, L2 is a linker unit attached to the side chain of aa, and each wavy line (~) represents a binding site for a Stretcher unit; 1 (poly) and aa 2 (Poly) is a polymer unit attached to each aa, each SU is a sugar unit attached to each aa; each CU is a carboxyl unit attached to each aa; double wavy line represents a binding site for at least one of said Drug units; 1 and aa 2 are independently selected from α, β and γ amino acids and derivatives thereof; 83. The linker compound of any one of claims 1 to 82.

88. 88. The linker compound of any one of claims 1 to 87, wherein the linker unit is a cleavable linker unit.

89. 89. The linker compound of claim 88, wherein the linker unit comprises a peptide that is cleavable by an intracellular protease.

90. 90. The linker compound of claim 89, wherein said 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.

91. 88. The linker compound of any one of claims 1-87, wherein said amino acid unit comprises a peptide that is cleavable by an intracellular protease.

92. 92. The linker compound of claim 91, wherein said 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.

93. 93. The linker compound of any one of claims 88-92, wherein the cleavable peptide is attached to a para-aminobenzyl alcohol self-immolative group (PABA).

94. The following structure: including one of In the formula, the wavy line on the oxygen group or *-amino group indicates the site of attachment to at least one of the Drug units or for a linking group attached to at least one of the Drug units; a wavy line on the amino group indicates a binding site for a Stretcher unit or an amino acid unit, or indicates H before binding, 94. The linker compound of any one of claims 1 to 93.

95. 95. The linker compound of any one of claims 1 to 94, wherein said linker unit further comprises a Stretcher unit having a binding site for a targeting unit, wherein said Stretcher unit is attached to said amino acid unit of said linker compound.

96. The stretcher unit is: is selected from the group consisting of: each indicates the binding site of the Stretcher unit to the amino acid unit; 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 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- (where b is 1 to 26), -C 1 ~C 8 Alkylene-(CH 2 -O-CH 2 ) b -C 1 ~C 8 alkylene-NH- (where b is 1 to 26), -C 1 ~C 8 Alkylene-(C(=O))-NH-(CH 2 -O-CH 2 ) b -C(=O)- (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- (where b is 1 to 26), -C 3 ~C 8 Carbocyclo-NH-, -O-(C 1 ~C 8 alkyl)-NH-, -arylene-NH-, -C 1 ~C 10 Alkylene-arylene-NH-, -arylene-C 1 ~C 10 Alkylene-NH-, -C 1 ~C 10 Alkylene-(C 3 ~C 8 carbocyclo)-NH-, -(C 3 ~C 8 Carbocyclo)-C 1 ~C 10 Alkylene-NH-, -C 3 ~C 8 Heterocyclo-NH-, -C 1 ~C 10 Alkylene-(C 3 ~C 8 heterocyclo)-NH-, -(C 3 ~C 8 Heterocyclo)-C 1 ~C 10 Alkylene-NH-, -C 1 ~C 10 Alkylene-S-, C 1 ~C 10 Heteroalkylene -S-, -C 3 ~C 8 Carbocyclo-S-, -O-(C 1 ~C 8 Alkyl)-S-, -arylene-S-, -C 1 ~C 10 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 unit 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; 96. The linker compound of claim 95.

97. The stretcher unit comprises: or a stereoisomer thereof, In the formula, each R a are independently H or C 1～6 alkyl, each n is independently 0 to 12, and a wavy line indicates the attachment site of the Stretcher unit to the Amino Acid unit, and the attachment site for the Targeting unit is on a maleimide, a primary amine, or an alkyne functional group; 96. The linker compound of claim 95.

98. The stretcher unit comprises: or a stereoisomer thereof, In the formula, wavy line indicates the attachment site of the Stretcher unit to an Amino Acid unit, and the attachment site for the Targeting unit is on a maleimide, a primary amine, or an alkyne functional group; 96. The linker compound of claim 95.

99. The following structure: or a stereoisomer thereof, In the formula, wavy line represents an attachment site to at least one of the Drug units or for a linking group attached to at least one of the Drug units; 99. The linker compound of any one of claims 1 to 98.

100. 100. A Drug-Linker Compound comprising the linker compound of any one of claims 1-99 conjugated to at least one Drug unit.

101. 101. The Drug-Linker of Claim 100, 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.

102. 102. The Drug-Linker of Claim 101, wherein the Drug unit is a cytotoxic agent.

103. 103. The drug-linker of claim 102, wherein said cytotoxic agent is selected from the group consisting of an auristatin, a maytansinoid, a camptothecin, a duocarmycin, and a calicheamicin.

104. The drug-linker of claim 103, wherein the cytotoxic agent is an auristatin.

105. The drug-linker of claim 104, wherein the cytotoxic agent is MMAE or MMAF.

106. The drug-linker of claim 103, wherein the cytotoxic agent is camptothecin.

107. The drug-linker of claim 106, wherein the cytotoxic agent is exatecan or SN-38.

108. The drug-linker of claim 107, wherein the cytotoxic agent is RS-exatecan or SS-exatecan.

109. The drug-linker of claim 103, wherein the cytotoxic agent is calicheamicin.

110. The drug-linker of claim 103, wherein the cytotoxic agent is a maytansinoid.

111. 111. The drug-linker of claim 110, wherein the maytansinoid is maytansine, maytansinol, or ansamatocin-2.

112. 102. The Drug-Linker of Claim 101, wherein the Drug unit is an immunomodulator.

113. The drug-linker of claim 112, wherein said immunomodulatory agent is selected from a TRL7 agonist, a TLR8 agonist, a STING agonist, or a RIG-I agonist.

114. The drug-linker of claim 113, wherein said immunomodulator is a TLR7 agonist.

115. The drug-linker of claim 114, 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.

116. The drug-linker of claim 113, wherein said immunomodulator is a TLR8 agonist.

117. The drug-linker of claim 116, 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.

118. The drug-linker of claim 113, wherein the immunomodulator is a STING agonist.

119. The drug-linker of claim 113, wherein the immunomodulator is a RIG-I agonist.

120. The drug-linker of claim 119, wherein the RIG-I agonist is selected from KIN1148, SB-9200, KIN700, KIN600, KIN500, KIN100, KIN101, KIN400 and KIN2000.

121. 102. The Drug-Linker of Claim 101, wherein said Drug unit is a chelating ligand.

122. 122. The drug-linker of claim 121, 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.

123. The following structure: or stereoisomers thereof.

124. 124. A conjugate comprising a targeting unit attached to the drug-linker of any one of claims 100 to 123, wherein the targeting unit specifically binds to a target molecule.

125. 125. The conjugate of claim 124, wherein the targeting unit is selected from an antibody or an antigen-binding portion thereof.

126. 126. The conjugate of claim 125, wherein the targeting unit is a monoclonal antibody, Fab, Fab', F(ab'), Fv, disulfide-linked Fc, scFv, single domain antibody, diabody, bispecific antibody, or multispecific antibody.

127. The conjugate of claim 124 or 125, wherein the targeting unit 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.

128. 125. The conjugate of claim 124, wherein said targeting unit is a diabody, DART, anticalin, affibody, avimer, DARPin, or adnectin.

129. 129. The conjugate of any one of claims 124-128, wherein said targeting unit is monospecific.

130. 130. The conjugate of any one of claims 124-129, wherein said targeting unit is bivalent.

131. 130. The conjugate of any one of claims 124-129, wherein said targeting unit is bispecific.

132. The average drug loading of the complex (p load 132. The conjugate of any one of claims 124-131, wherein q 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.

133. below:

133. The conjugate of any one of claims 124-132, selected from:

134. 134. The conjugate of claim 133, wherein the target molecule is CD19, CD20, CD30, CD33, CD70, LIV-1, HER2, or EGFRv3.

135. 135. The conjugate of any one of claims 124-134, wherein the target molecule is a cancer-associated antigen.

136. The target molecule is CD19, CD20, CD30, CD33, CD38, CA125, 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), HER2, 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, TUBB3, VH L, CDH1, ERBB4, FBXW7, HNF1A, JAK3, NPM1, PTPN11, RB1, SMAD4, SMARCB1, STK1, MLH1, MSH2, MSH6, PMS2, ROS1, ERCC1, 5T4(TPBG), B7-H3, CCR7, CD105, CD2 2, 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.

137. The targeting unit is selected from the group consisting of rituximab (Rituxan®), trastuzumab (Herceptin®), pertuzumab (Perjeta®), bevacizumab (Avastin®), ranibizumab (Lucentis®), cetuximab (Erbitux®), alemtuzumab (Campath®), panitumumab (Vectibix®), ibritumomab 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®), hu The conjugate of any one of claims 124-126-130, 131, or 132, which is an antibody or fragment thereof comprising an anti-SLITRK6 antibody comprising 1H2-03, or an anti-LHRH receptor antibody comprising clone A9E4, F1G4, AT2G7, GNRH03, or GNRHR2.

138. 138. A pharmaceutical composition comprising the conjugate of any one of claims 124-137 and a pharmaceutically acceptable carrier.

139. 1. A method of treating a subject in need thereof, comprising: administering to said subject a conjugate of any one of claims 124-137 or a pharmaceutical composition of claim 138; 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.

140. formula: R 0 -(R 1 -[CH 2 -CH(OH)-CH 2 -O] n0 -R 6 ) n2 (XXXXI); or R 0 -(R 1 -[[CH 2 -CH 2 -O] n0 -R 2 -NR 4 -R 5 ) n2 (XXXXII); or A polar group represented by a salt of any one of them, During the ceremony, R 0 has the following structure: having one of: Each R 1 are independently a bond or C 1 ~C 3 an alkylene group; R 2 is independently C 1 ~C 3 alkylene groups; R 4 and R 5 are each independently selected from H or a polyhydroxyl group, where R 4 and R 5 Both of these cannot be H; Each R 6 is independently C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkylene-NR 7 R 8 , -C(O)N(R N )-C 1 ~C 6 Alkylene-NR 4 R 5 and R N is H or C 1 ~C 4 is alkyl; R 7 and R 8 are each independently selected from H, a polyhydroxyl group, or a —C(O)-polyhydroxyl group; each n0 is independently 1 to 26; n2 is 1 or 2, The polar group.

141. formula: R 0 -(R 1 -[CH 2 -CH(OH)-CH 2 -O] n0 -R 6 ) n2 (XXXXI); is represented by During the ceremony, R 0 has the following structure: having one of: Each R 1 are independently a bond or C 1 ~C 3 an alkylene group; R 4 and R 5 are each independently selected from H or a polyhydroxyl group, where R 4 and R 5 Both of these cannot be H; Each R 6 is independently C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkylene-NR 7 R 8 , -C(O)N(R N )-C 1 ~C 6 Alkylene-NR 4 R 5 and R N is H or C 1 ~C 4 is alkyl; R 7 and R 8 are each independently selected from H, a polyhydroxyl group, or a —C(O)-polyhydroxyl group; each n0 is independently 1 to 26; n2 is 1 or 2, 141. The polar group of claim 140.

142. R 0 but 142. The polar group of claim 140 or 141, wherein:

143. R 0 but 142. The polar group of claim 140 or 141, wherein:

144. R 1 144. The polar group of any one of claims 140-143, wherein is a bond.

145. 145. The polar group of any one of claims 140-144, wherein n0 is 4 to 10.

146. 146. The polar group of any one of claims 140-145, wherein n0 is 6.

147. R 6 C 1 ~C 6 Alkylene-C(OH)HC 1 ~C 6 Alkylene-NR 7 R 8 147. The polar group of any one of claims 140 to 146, wherein

148. R 6 C 1 -AlkyleneC(OH)HC 1 -Alkylene-NR 7 R 8 148. The polar group of any one of claims 140-147, wherein

149. R 7 is a polyhydroxyl group.

150. R 8 150. The polar group of any one of claims 140-149, wherein

151. R 6 -C(O)N(R N )-C 1 ~C 6 Alkylene-NR 4 R 5 146. The polar group of any one of claims 140 to 145,

152. R 4 152. The polar group of any one of claims 140-145, or 151, wherein is a polyhydroxyl group.

153. R 5 152. The polar group of any one of claims 140-145, or 151, wherein is a polyhydroxyl group.

154. R N 154. The polar group of any one of claims 140-145, or 151-153, wherein

155. 155. The polar group of any one of claims 140-145, or 151-154, wherein n2 is 2.

156. A polar group according to claim 140 selected from:

157. A polar group according to claim 140 selected from:

158. formula: R 0 -(R 1 -[[CH 2 -CH 2 -O] n0 -R 2 -NR 4 -R 5 ) n2 (XXXXII) 2. The polar group of claim 1, represented by:

159. R 0 but 159. The polar group of claim 158, wherein:

160. R 1 160. The polar group of any one of claims 158-159, wherein is a bond.

161. 161. The polar group of any one of claims 158-160, wherein n0 is 6-10.

162. 162. The polar group of any one of claims 158-161, wherein n0 is 8.

163. R 2 independently C 1 ~C 3 163. The polar group of any one of claims 158-162, selected from an alkylene group.

164. R 4 and R 5 is a polyhydroxyl group.

165. 165. The polar group of any one of claims 158-164, wherein n2 is 2.

166. A polar group according to claim 140 selected from:

166. A polar group according to claim 140 selected from:

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