Protease / Enzyme Cleavable Linker-Payload and Protein Conjugate

Abstract

Compounds, their conjugate products, methods, and pharmaceutical compositions for use in treatment and diagnosis are provided herein.

Description

Technical Field

[0001] Cross - reference to related applications

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 389,666, filed on July 15, 2022, which is hereby incorporated by reference in its entirety for all purposes.

[0002]

[0002] The present disclosure relates to linker - payload compounds and their polymeric conjugates; pharmaceutical compositions comprising linker - payload compounds and / or conjugates; methods of producing linker - payload compounds and / or conjugates; and methods of using linker - payload compounds, conjugates, and compositions, inter alia, for the treatment of cell proliferative disorders including cancer.

Background Art

[0003]

[0003] Biopharmaceuticals offer rich treatment and diagnostic possibilities to patients worldwide. However, many drugs based on macromolecules such as proteins, peptides, and antibodies present limitations to their effective use, including limitations in bioavailability, absorption, distribution, metabolism, and excretion (ADME). Some of these limitations can affect drug dosage, half - life, side effects, and toxicity. Strategies for improving the effectiveness of biopharmaceuticals are still needed.

Summary of the Invention

Means for Solving the Problems

[0004]

[0004] In one aspect, a compound of formula (I):

[0005]

Chemical Formula

[0006] [Chemistry] selected from; Ring A is an optionally substituted bridged, fused, or spiro bicyclic carbocyclic ring, or an optionally substituted bridged, fused, or spiro bicyclic heterocyclic ring, and the carbocyclic or heterocyclic ring of Ring A is alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)-, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl, and may be substituted with one or more substituents selected therefrom; Ring B is an optionally substituted N-bonded bridged, fused, or spiro bicyclic heterocyclic ring, and Ring B is alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)-, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl, and may be substituted with one or more substituents selected therefrom; R a and R b are independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; a is an integer independently selected from 0, 1, 2, 3, 4, 5, and 6; b is an integer selected from 0 and 1; R 1 is alkyl optionally substituted with one or more substituents selected from hydrogen, or cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aryl, and heteroaryl; R 2 and R 3 are independently selected from hydrogen, alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; Y is *-C(O)-(CR a R b ) c -NH- or *-C(O)-(CR a R b ) c -, where * represents the location where Y is attached to RG; c is an integer selected from 1, 2, 3, 4, 5, and 6; RG is a reactive group; L 2 is either absent or a linker containing a hydrophilic polymer residue; L 3 is absent, -C(O)-AA-, -C(O)-AA-Z-(CR a R b ) a -Z-(CR a R b ) a -C(O)-, -C(O)-Z-(CR a R b ) a -C(O)-Z-L 4 -OC(O)-, -Z-AA-, -AA-, -C(O)-, -C(O)-AA-Z-(CR a R b ) a-, -AA-C(O)-, -C(O)-(CR a R b ) a -Z-(CR a R b ) a -Z-AA-C(O)-, -C(O)O-L 4 -Z-C(O)-(CR a R b ) a -Z-C(O)-, -AA-Z-, or -(CR a R b ) a -Z-AA-C(O)-; Z is selected from -NR 2 - and -O-; AA is an amino acid residue or a peptide residue; L 4 is

[0007]

Chem.

[0008]

Chem.

[0009]

[0005] In certain embodiments, the compound of formula (I) is a compound of formula (IA):

[0010]

Chem.

[0011]

[0006] In certain embodiments, the compound of formula (I) is a compound of formula (IB):

[0012]

Chemical Structure

[0013]

[0007] In certain embodiments, the compound of formula (I) is a compound of formula (IC):

[0014]

Chemical Structure

[0015]

[0008] In certain embodiments, the compound of formula (I) is a compound of formula (ID):

[0016]

Chemical Structure

[0017]

[0009] In certain embodiments, the compound of formula (I) is a compound of formula (IE):

[0018]

Chemical formula

[0019]

[0010] In certain embodiments, the compound of formula (I) is a compound of formula (IF):

[0020]

Chemical formula

[0021]

[0011] In certain embodiments, the compound of formula (I) is a compound of formula (IG):

[0022]

Chemical formula

[0023]

[0012] In certain embodiments, the compound of formula (I) is a compound of formula (IH):

[0024]

Chemical formula

[0025]

[0013] In one aspect, the conjugate of formula (II):

[0026]

Chemical formula

[0027]

Chemical formula

[0028]

Chemical formula

[0029]

Chemical formula

[0030]

[0014] In certain embodiments, the compound of formula (II) is a compound of formula (IIA):

[0031]

Chemical formula

[0032]

[0015] In certain embodiments, the compound of formula (II) is a compound of formula (IIB):

[0033]

Chemical formula

[0034]

[0016] In certain embodiments, the compound of formula (II) is a compound of formula (IIC):

[0035]

Chemical formula

[0036]

[0017] In certain embodiments, the compound of formula (II) is a compound of formula (IID):

[0037]

Chemical formula

[0038]

[0018] In certain embodiments, the compound of formula (II) is a compound of formula (IIE):

[0039]

Chem.

[0040]

[0019] In certain embodiments, the compound of formula (II) is a compound of formula (IIF):

[0041]

Chem.

[0042]

[0020] In certain embodiments, the compound of formula (II) is a compound of formula (IIG):

[0043]

Chem.

[0044] In certain embodiments, the compound of formula (II) is a compound of formula (IIH):

[0045]

Chemical formula

[0046] In one aspect, a conjugate of formula (III):

[0047]

Chemical formula

[0048] Non-limiting non-natural amino acids include sulfonalanine, hydroxyproline (Hyp), β-alanine, citrulline (Cit), ornithine (Orn), norleucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, and methionine sulfone.

[0049] In certain embodiments, the compound of formula (III) is a compound of formula (IIIA):

[0050] [Chemical formula] or a pharmaceutically acceptable salt and / or stereoisomer thereof (wherein RG, Y, L 2 , L 3 , and D are as defined herein) is.

[0051]

[0025] In certain embodiments, the compound of formula (III) is a compound of formula (IIIB):

[0052] [Chemical formula] or a pharmaceutically acceptable salt and / or stereoisomer thereof (wherein integer a, integer c, RG, R a , R b , POLY 1 , AA, and D are as defined herein) is.

[0053]

[0026] In certain embodiments, the compound of formula (IIIB) is of the formula:

[0054] [Chemical formula] a compound of or a pharmaceutically acceptable salt and / or stereoisomer thereof (wherein integer a, integer c, RG, R a , R b , POLY 1 , AA, and D are as defined herein) is.

[0055]

[0027] In one aspect, the conjugate of formula (IV):

[0056] [Chemical formula] or a pharmaceutically acceptable salt and / or stereoisomer thereof (wherein L 5 is a linker containing a non-natural amino acid; RL, COMP, Y, L 2 L 3 , and D are as defined herein) is provided herein.

[0057]

[0028] In certain embodiments, the compound of formula (IV) is a compound of formula (IVA):

[0058]

Chemical formula

[0059]

[0029] In certain embodiments, the compound of formula (IV) is a compound of formula (IVB):

[0060]

Chemical formula

[0061]

[0030] In certain embodiments, the compound of formula (IVB) is of the formula:

[0062] [Chemistry] The compound of or its pharmaceutically acceptable salt and / or positional isomer (wherein the integer a, integer c, RL, COMP, R a 、R b 、POLY 1 、AA, and D are as defined herein) is.

[0063]

[0031] The present disclosure provides at least the following embodiments: a) A compound of formula (I), formula (IA), formula (IB), formula (IC), formula (ID), formula (IE), formula (IF), formula (IG), formula (IH), formula (IIIA), or formula (IIIB), or a pharmaceutically acceptable salt, solvate, positional isomer, and / or stereoisomer thereof; b) A compound selected from Compounds 1 to 25, or a pharmaceutically acceptable salt, solvate, positional isomer, and / or stereoisomer thereof; c) A pharmaceutical composition comprising the compound of (a) or (b) and a pharmaceutically acceptable excipient, diluent, or carrier; d) A conjugate of formula (II), formula (IIA), formula (IIB), formula (IIC), formula (IID), formula (IIE), formula (IIF), formula (IIG), formula (IIH), formula (IVA), or formula (IVB), or a pharmaceutically acceptable salt, solvate, positional isomer, and / or stereoisomer thereof; e) A compound selected from Compounds 101A to 125B, or a pharmaceutically acceptable salt, solvate, positional isomer, and / or stereoisomer thereof; f) A pharmaceutical composition comprising the compound of (d) or (e) and a pharmaceutically acceptable excipient, diluent, or carrier; g) A method of treating a disease or disorder in a subject in need thereof, the method comprising administering a therapeutically effective amount of the compound of (a) or (b) or the pharmaceutical composition of (c); h) A method for treating a disease or disorder in a subject in need thereof, the method comprising administering a therapeutically effective amount of the compound of (d) or (e) or the pharmaceutical composition of (f); i) A method for inhibiting tubulin polymerization in a subject in need thereof, the method comprising administering a therapeutically effective amount of the compound of (a), (b), (d), or (e) or the pharmaceutical composition of (c) or (f); j) The method of (g) or (h), wherein the disease or disorder is abnormal cell proliferation; k) The method of (j), wherein the abnormal cell proliferation is cancer; l) The method of (k), wherein the cancer is small cell lung cancer, non-small cell lung cancer, ovarian cancer, platinum-resistant ovarian cancer, ovarian adenocarcinoma, endometrial cancer, breast cancer, breast cancer overexpressing HER2, triple-negative breast cancer, lymphoma, large cell lymphoma, diffuse mixed histiocytic and lymphocytic lymphoma, follicular B cell lymphoma, colon cancer, colorectal cancer, colon adenocarcinoma, colorectal adenocarcinoma, melanoma, prostate cancer, or multiple myeloma; m) Use of a therapeutically effective amount of the compound of (a) or (b) or the pharmaceutical composition of (c) in the treatment of a disease or disorder in a subject in need thereof; n) Use of a therapeutically effective amount of the compound of (d) or (e) or the pharmaceutical composition of (f) in the treatment of a disease or disorder in a subject in need thereof; o) Use of a therapeutically effective amount of the compound of (a) or (b) or the pharmaceutical composition of (c) in the manufacture of a medicament for the treatment of a disease or disorder in a subject in need thereof; p) Use of a therapeutically effective amount of the compound of (d) or (e) or the pharmaceutical composition of (f) in the manufacture of a medicament for the treatment of a disease or disorder in a subject in need thereof; q) Use of a therapeutically effective amount of the compound of (a), (b), (d), or (e) or the pharmaceutical composition of (d) or (f) in the manufacture of a medicament for inhibiting tubulin polymerization in a subject in need thereof; r) The use of (m)-(p), wherein the disease or disorder is abnormal cell proliferation; s) The use of (r), wherein the abnormal cell proliferation is cancer; (t) the use of a cancer being small cell lung cancer, non-small cell lung cancer, ovarian cancer, platinum-resistant ovarian cancer, ovarian adenocarcinoma, endometrial cancer, breast cancer, breast cancer overexpressing HER2, triple-negative breast cancer, lymphoma, large cell lymphoma, diffuse mixed histiocytic and lymphocytic lymphoma, follicular B cell lymphoma, colon cancer, colorectal cancer, colon adenocarcinoma, colorectal adenocarcinoma, melanoma, prostate cancer, or multiple myeloma; and (u) A method for producing a conjugate of formula (II), formula (IIA), formula (IIB), formula (IIC), formula (IID), formula (IIE), formula (IIF), formula (IIG), formula (IIH), formula (IVA), or formula (IVB), or a pharmaceutically acceptable salt thereof, the method comprising contacting a compound of formula I, formula (IA), formula (IB), formula (IC), formula (ID), formula (IE), formula (IF), formula (IG), formula (IH), formula (IIIA), or formula (IIIB) with a second compound under conditions suitable for conjugating the compound of formula (I), formula (IA), formula (IB), formula (IC), formula (ID), formula (IE), formula (IF), formula (IG), formula (IH), formula (IIIA), or formula (IIIB) with the second compound, wherein the second compound comprises an alkyne, cyclooctyne, strained alkene, tetrazine, methylcyclopropene, thiol, maleimide, carbonyl, amine, oxyamine, or azide.

Brief Description of the Drawings

[0064]

Figure 1A

[0032] A graph showing the killing activity of anti-FolRα ADC conjugated with Compound 1, Compound 2, and Compound 3 against FolRα-positive Igrov1 cells as described in Example 18.

Figure 1B

[0033] A graph showing that anti-FolRα ADC conjugated with Compound 1, Compound 2, and Compound 3 does not show killing activity against FolRα-negative A549 cells as described in Example 18.

Figure 2A

[0034] As described in Example 18, it is a graph showing the killing activity of anti-FolRα ADC conjugated with Compound 6, Compound 7, Compound 8, and Compound 9 against FolRα-positive Igrov1 cells.

Figure 2B

[0035] As described in Example 18, it is a graph showing that anti-FolRα ADC conjugated with Compound 6, Compound 7, Compound 8, and Compound 9 does not show killing activity against hFolRα-negative A549 cells.

Figure 3A

[0036] As described in Example 19, it is a graph showing the killing activity of anti-FolRα ADC conjugated with Compound 13, Compound 14, and Compound 15 against FolRα-positive Igrov1 cells.

Figure 3B

[0037] As described in Example 19, it is a graph showing that anti-FolRα ADC conjugated with Compound 13, Compound 14, and Compound 15 does not show killing activity against FolRα-negative A549 cells.

Mode for Carrying Out the Invention

[0065]

[0038] For example, compounds of formula (I), formula (IA), formula (IB), formula (IC), formula (ID), formula (IE), formula (IF), formula (IG), formula (IH), formula (IIA), and formula (IIB) that are useful for modulating the bioavailability and ADME of a polymeric conjugate compound are described herein. In some cases, the compounds described herein are useful for preparing conjugates for in vivo use, such as conjugates of formula (II), formula (IIA), formula (IIB), formula (IIC), formula (IID), formula (IIE), formula (IIF), formula (IIG), formula (IIH), formula (IVA), or formula (IVB). In certain embodiments, the compounds and conjugates are characterized by functional groups suitable for enzymatic cleavage to release a payload compound for in vivo or other use. These compounds can be varied to adjust the physiochemical properties and plasma stability of the conjugate. Thereby, a platform for modulating the bioavailability and ADME of polymers in vivo is provided.

[0066]

[0039] Definitions Unless otherwise defined, all technical terms, notations, and other scientific terms used in this specification are intended to have the meanings commonly understood by those of ordinary skill in the art to which this disclosure belongs. In some instances, terms with commonly understood meanings are defined in this specification for clarity and / or for ready reference. The techniques and procedures described or referenced herein are generally well understood and commonly employed by those of ordinary skill in the art using conventional methodologies, such as those described in Green & Sambrook, Molecular Cloning: A Laboratory Manual, 4th Edition (2012), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; and Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons. Procedures involving the use of commercially available kits and reagents are generally carried out according to the protocols and conditions defined by the manufacturer, unless otherwise noted.

[0067]

[0040] As used in this specification, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

[0068]

[0041] The term “about” refers to the indicated value as well as the ranges above and below that value, including those values. In certain embodiments, the term “about” indicates ±10%, ±5%, or ±1% of the specified value. In certain embodiments, the term “about” indicates ± a certain standard deviation of the specified value. In certain embodiments, for example, on a logarithmic scale (e.g., pH), the term “about” indicates ±0.3, ±0.2, or ±0.1 of the specified value.

[0069] When referring to the compounds provided in this specification, unless otherwise indicated, the following terms have the following meanings. Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. If there are multiple definitions for a term in this specification, unless otherwise explicitly stated, the definitions in this section shall prevail.

[0070]

[0043] As used herein, the terms "alkoxy" and "alkoxyl" refer to an -OR’’ group, wherein R’’ is alkyl or cycloalkyl. In certain embodiments, alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

[0071]

[0044] As used herein, the term "alkoxyamine" refers to an -alkylene-O-NH2 group, wherein alkylene is as defined herein. In some embodiments, the alkoxyamine group can react with an aldehyde to form an oxime residue. Examples of alkoxyamine groups include -CH2CH2-O-NH2, -CH2-O-NH2, and -O-NH2.

[0072]

[0045] As used herein, the term "alkyl" refers to a saturated straight-chain or branched hydrocarbon, unless otherwise specified. In certain embodiments, the alkyl group is a primary, secondary, or tertiary hydrocarbon. In certain embodiments, the alkyl group contains 1 to 10 carbon atoms (i.e., C1-C 10 alkyl). In certain embodiments, alkyl is lower alkyl, e.g., C 1~6Such as alkyl. In certain embodiments, the alkyl group is selected from the group consisting of methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl. In certain embodiments, "substituted alkyl" refers to an alkyl substituted with, for example, one, three, or three groups independently selected from halogen (e.g., fluoro (F), chloro (Cl), bromo (Br), or iodo (I)), alkyl, -CN, -NO2, amide, -C(O)-, -C(S)-, ester, carbamate, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, dialkylamino, haloalkyl, hydroxyl, amino, alkylamino, and alkoxy. In some embodiments, the alkyl is unsubstituted.

[0073]

[0046] As used herein, the term "alkylene" refers to a divalent alkyl group as defined herein, unless otherwise specified. "Substituted alkylene" refers to an alkylene group substituted as described herein for alkyl. In some embodiments, the alkylene is unsubstituted.

[0074]

[0047] As used herein, in certain embodiments, the term "alkenyl" refers to an olefinically unsaturated hydrocarbon group having up to about 11 carbon atoms or 2 to 6 carbon atoms (e.g., "lower alkyl"), which may be straight-chain or branched, and having at least one or one to two olefinically unsaturated sites. "Substituted alkenyl" refers to an alkenyl group substituted as described herein for alkyl.

[0075]

[0048] As used herein, the term "alkenylene" refers to a divalent alkenyl as defined herein. Lower alkenylene is, for example, C2-C6-alkenylene.

[0076]

[0049] As used herein, the term "alkynyl" in certain embodiments refers to an acetylenically unsaturated hydrocarbon group having up to about 11 carbon atoms or 2 to 6 carbon atoms (e.g., "lower alkynyl"), which may be linear or branched, and having at least one or one to two acetylenic unsaturation sites. Non-limiting examples of alkynyl groups include acetylene (-C≡CH), propargyl (-CH2C≡CH), and the like. "Substituted alkynyl" refers to an alkynyl group substituted as described herein for alkyl.

[0077]

[0050] As used herein, the term "alkynylene" refers to a divalent alkynyl as defined herein. Lower alkynylene is, for example, C2-C6-alkynylene.

[0078]

[0051] As used herein, the term "amino" refers to -NH2.

[0079]

[0052] As used herein, the term "alkylamino", unless otherwise specified, refers to an -NHR'' group (wherein R'' is, for example, Calkyl, Calkenyl, Calkynyl, Ccarbocyclic, 3- to 12-membered heterocyclic, C haloalkyl, etc. as defined herein). In certain embodiments, the alkylamino is Calkylamino. 1~10 alkyl, C 2~10 alkenyl, C 2~10 alkynyl, C 3~12 carbocyclic, 3- to 12-membered heterocyclic, C 1~10 haloalkyl, etc.). In certain embodiments, the alkylamino is C 1~6 alkylamino.

[0080]

[0053] As used herein, the term "dialkylamino", unless otherwise specified, refers to an -NR''R'' group (wherein each R'' is independently Calkyl as defined herein). In certain embodiments, the dialkylamino is, for example, di-C 1~10 alkylamino, C 1~6 alkylamino, C 2~10 alkenyl, C 2~10 alkynyl, C 3~12 carbocyclic, 3- to 12-membered heterocyclic, C1~10 It is a haloalkyl or the like.

[0081]

[0054] As used herein, the term "aryl", unless otherwise specified, refers to phenyl, biphenyl, or naphthyl. This term includes both substituted and unsubstituted moieties. An aryl group may be substituted with one or more moieties (e.g., in some embodiments, one, two, or three moieties) selected from the group consisting of, but not limited to, halogen (e.g., fluoro (F), chloro (Cl), bromo (Br), or iodo (I)), alkyl, haloalkyl, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, and phosphonate, each moiety being independently unprotected or protected as recognized by one of ordinary skill in the art (e.g., see Greene et al., Protective Groups in Organic Synthesis, John Wiley and Sons, 2nd Edition, 1991), and the aryl of arylamino and aryloxy substituents is further unsubstituted.

[0082]

[0055] As used herein, the term "arylamino", unless otherwise specified, refers to an -NR’R’’ group (wherein R’ is hydrogen or C1-C6-alkyl; and R’’ is aryl as defined herein).

[0083]

[0056] As used herein, the term "arylene", unless otherwise specified, refers to a divalent aryl group as defined herein.

[0084]

[0057] As used herein, the term "aryloxy", unless otherwise specified, refers to an -OR group (wherein R is aryl as defined herein).

[0085]

[0058] "Alkarylene" refers to an arylene group as defined herein, wherein the aryl ring is substituted with one or two alkyl groups. "Substituted alkarylene" refers to an alkarylene as defined herein, wherein the arylene group is further substituted as defined herein for aryl.

[0086]

[0059] "Aralkylen" refers to a -CH2-arylene-, -arylene-CH2-, or -CH2-arylene-CH2-group (wherein arylene is as defined herein). "Substituted aralkylen" refers to an aralkylen as defined herein, wherein the aralkylen group is substituted as defined herein for aryl.

[0087]

[0060] The term "carboxyl" or "carboxy" when used herein refers to -C(O)OH or -COOH.

[0088]

[0061] The term "cycloalkyl" or "carbocyclic" when used herein, unless otherwise specified, refers to a saturated, unsaturated, or aromatic ring in which all atoms of the ring are carbon. In certain embodiments, the "cycloalkyl" or "carbocyclic" group can be a saturated, and / or bridged, and / or unbridged, and / or fused bicyclic group, and / or a spiro bicyclic group. In certain embodiments, the "cycloalkyl" or "carbocyclic" group contains 3 to 10 carbon atoms (i.e., C3-C 10 cycloalkyl). In some embodiments, "cycloalkyl" or "carbocyclic" is 3 to 15 carbons (C 3~15 ), 3 to 10 carbons (C 3~10 ), 3 to 7 carbons (C 3~7 ), or 3 to 6 carbons (C 3~6) having (i.e., "lower cycloalkyl"). In certain embodiments, a "cycloalkyl" or "carbocyclic" group is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexylmethyl, cycloheptyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, decalinyl, or adamantyl. Exemplary "cycloalkyl" or "carbocyclic" groups include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl. "Cycloalkyl" or "carbocyclic" includes 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring of a bicyclic cycloalkyl or carbocyclic can be selected from saturated, unsaturated, and aromatic rings. A bicyclic cycloalkyl or carbocyclic includes any combination of saturated, unsaturated, and aromatic bicyclic rings as valence permits. A bicyclic cycloalkyl or carbocyclic includes any combination of ring sizes such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. Non-limiting examples of bridged bicyclic cycloalkyl or carbocyclic groups include, but are not limited to, bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, bicyclo[2.1.1]hexyl, bicyclo[3.1.1]heptyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl, bicyclo[3.3.2]decyl, and 2-oxabicyclo[2.2.2]octyl. Non-limiting examples of spirocyclic cycloalkyl or carbocyclic groups include, but are not limited to, spiro[3.3]heptyl, spiro[3.4]octyl, spiro[3.5]nonyl, spiro[3.6]decyl, spiro[4.4]nonyl, spiro[4.5]decyl, spiro[5.5]undecyl, spiro[5.6]dodecyl, and spiro[5.7]tridecyl.

[0089] The term "bicyclic ring system" includes a 6- to 12- (e.g., 8- to 12- or 9, 10, or 11-) membered structure forming two rings, the two rings having at least one common atom (e.g., two common atoms). The bicyclic ring can be fused, bridged, or spirocyclic. Bicyclic ring systems include bicyclic aliphatics (e.g., bicycloalkyl or bicycloalkenyl), bicyclic heteroaliphatics, bicyclic aryls, and bicyclic heteroaryls.

[0090]

[0063] The term "bridged bicyclic ring system" refers to a bicyclic heterocyclic aliphatic ring system or a bicyclic alicyclic ring system in which the rings are bridged. Examples of bridged bicyclic ring systems include, but are not limited to, adamantanyl, norbornanyl, bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, bicyclo[2.1.1]hexyl, bicyclo[3.1.1]heptyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl, bicyclo[3.3.2]decyl, 2-oxabicyclo[2.2.2]octyl, 6-azabicyclo[3.1.1]heptyl, 6-azabicyclo[3.1.1]heptyl, 1-azabicyclo[2.2.1]heptyl, 2-azabicyclo[2.2.1]heptyl, 7-azabicyclo[2.2.1]heptyl, 1-azabicyclo[2.2.2]octyl, 3-azabicyclo[3.2.1]octyl, and 2-oxabicyclo[3.1.1]heptyl, 2,6-dioxa-tricyclo[3.3.1.0 3,7Examples include nonyl. The bridged bicyclic ring system may be optionally substituted with one or more substituents such as alkyl (including carboxyalkyl, hydroxyalkyl, and haloalkyl such as trifluoromethyl), alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl, heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino, cycloalkylcarbonylamino, (cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino, (heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino, heteroaralkylcarbonylamino, cyano, halo, hydroxy, acyl, mercapto, alkylsulfanyl, sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, or carbamoyl.

[0091]

[0064] The term "spirobicyclic ring system" refers to a bicyclic heterocyclic aliphatic ring system or a bicyclic alicyclic ring system in which two or three rings are linked by one common atom. The spiro compounds represented by the overlapping rings indicate that the rings can be joined at any vertex. For example, the spiro group

[0092]

Chemical formula

[0093]

[0065] The term "cycloalkylene" as used herein refers to a divalent cycloalkyl group as defined herein. In certain embodiments, the cycloalkylene group is cyclopropylene

[0094]

Chem.

[0095]

Chem.

[0096]

Chem.

[0097]

Chem.

[0098]

Chem.

[0099]

[0066] The term "cycloalkylalkyl", as used herein, unless otherwise specified, refers to an alkyl group as defined herein substituted with one or two cycloalkyls as defined herein.

[0100]

[0067] The term "ester", as used herein, refers to -C(O)OR or -COOR (wherein R is an alkyl as defined herein).

[0101]

[0068] The term "fluorene", as used herein, may be substituted with one or more chemical functional groups described herein on any one or more carbons having one or more hydrogens,

[0102]

Chem.

[0103]

[0069] The term "haloalkyl" refers to an alkyl group as defined herein that is independently selected and substituted with one or more halogen atoms (e.g., in some embodiments, 1, 2, 3, 4, or 5).

[0104] As used herein, the term "heteroalkyl" refers to an alkyl as defined herein in which one or more carbon atoms are replaced by heteroatoms. As used herein, "heteroalkenyl" refers to an alkenyl as defined herein in which one or more carbon atoms are replaced by heteroatoms. As used herein, "heteroalkynyl" refers to an alkynyl as defined herein in which one or more carbon atoms are replaced by heteroatoms. Suitable heteroatoms include, but are not limited to, nitrogen (N), oxygen (O), and sulfur (S) atoms. Heteroalkyl, heteroalkenyl, and heteroalkynyl may be substituted. Examples of heteroalkyl moieties include, but are not limited to, aminoalkyl, sulfonylalkyl, and sulfinylalkyl. Examples of heteroalkyl moieties also include, but are not limited to, methylamino, methylsulfonyl, and methylsulfinyl. "Substituted heteroalkyl" refers to a heteroalkyl substituted with one, two, or three groups independently selected from halogen (e.g., fluoro (F), chloro (Cl), bromo (Br), or iodo (I)), alkyl, haloalkyl, hydroxyl, amino, alkylamino, and alkoxy. In some embodiments, a heteroalkyl group may contain one, two, three, or four heteroatoms. One of ordinary skill in the art will recognize that a 4-membered heteroalkyl may generally contain one or two heteroatoms, a 5- or 6-membered heteroalkyl may generally contain one, two, or three heteroatoms, and a 7- to 10-membered heteroalkyl may generally contain one, two, three, or four heteroatoms.

[0105]

[0071] As used herein, the term "heteroalkylene" refers to a divalent heteroalkyl as defined herein. "Substituted heteroalkylene" refers to a divalent heteroalkyl as defined herein substituted as described for heteroalkyl.

[0106]

[0072] The term "heterocycloalkyl" or "heterocycle" refers to a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms, and nitrogen or sulfur atoms may optionally be oxidized, and nitrogen atoms may optionally be quaternized, and the remaining ring atoms of the non-aromatic ring are carbon atoms. "Heterocycloalkyl" or "heterocycle" includes 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. In certain embodiments, "heterocycloalkyl" or "heterocycle" is a monovalent, monocyclic, or polycyclic fully saturated ring system. In certain embodiments, the "heterocycloalkyl" or "heterocycle" group can be an unsaturated, and / or bridged, and / or unbridged, and / or fused bicyclic group, and / or a spiro bicyclic group. The bicyclic "heterocycloalkyl" or "heterocycle" includes any combination of ring sizes such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. In certain embodiments, the "heterocycloalkyl" or "heterocycle" group has 3 to 20, 3 to 15, 3 to 10, 3 to 8, 4 to 7, 4 to 11, or 5 to 6 ring atoms. "Heterocycloalkyl" or "heterocycle" can be attached to the core structure by any heteroatom or carbon atom that results in the creation of a stable compound. In certain embodiments, "heterocycloalkyl" or "heterocycle" may include a fused ring system or a bridged ring system or a spiro ring system, and nitrogen or sulfur atoms may optionally be oxidized, and / or nitrogen atoms may optionally be quaternized, and is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system.In some embodiments, the "heterocycloalkyl" or "heterocyclic" group includes, but is not limited to, 2,5-diazabicyclo[2.2.2]octanyl, decahydroisoquinolinyl, dihydrobenzisoxazinyl, dihydrofuryl, dihydroisoindolyl, dihydropyranyl, dihydropyrazolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl, dihydropyrrolyl, dioxolanyl, 1,4-dithianyl, furanonyl, imidazolidinyl, imidazolinyl, indolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, oxazolidinonyl, oxazolidinyl, oxiranyl, piperazinyl, piperidinyl, 4-piperidonyl, pyrazolidinyl, pyrazolinyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl, tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothienyl, thiomorpholinyl, thiazolidinyl, tetrahydroquinolinyl, and 1,3,5-trithianyl. Non-limiting examples of bridged heterocycloalkyl or heterocyclic groups include, but are not limited to, 6-azabicyclo[3.1.1]heptyl, 6-azabicyclo[3.1.1]heptyl, 1-azabicyclo[2.2.1]heptyl, 2-azabicyclo[2.2.1]heptyl, 7-azabicyclo[2.2.1]heptyl, 1-azabicyclo[2.2.2]octyl, 3-azabicyclo[3.2.1]octyl, and 2-oxabicyclo[3.1.1]heptyl, 2,6-dioxa-tricyclo[3.3.1.0. 3,7 nonyl. Non-limiting examples of spirocyclic heterocycloalkyl or heterocyclic groups include, but are not limited to, 2,8-diazaspiro[4.5]decyl; 2,7-diazaspiro[3.5]nonyl; 3,9-diazaspiro[5.5]undecyl; 3-azaspiro[5.5]undecyl; 2-oxa-6-azaspiro[3.4]octyl; 2-oxa-9-azaspiro[5.5]undecyl; 3-oxa-9-azaspiro[5.5]undecyl; 7-azaspiro[3.5]nonyl; 2-azaspiro[3.5]nonyl; 7-oxaspiro[3.5]nonyl; and 2-oxaspiro[3.5]nonyl.

[0107]

[0073] In certain embodiments, "heterocycloalkyl" or "heterocycle" may also be optionally substituted as described herein. In certain embodiments, "heterocycloalkyl" or "heterocycle" is substituted with one, two, or three groups independently selected from halogen (e.g., fluoro (F), chloro (Cl), bromo (Br), or iodo (I)), alkyl, haloalkyl, hydroxyl, amino, alkylamino, and alkoxy. In some embodiments, the heterocycloalkyl or "heterocycle" group may contain one, two, three, or four heteroatoms. One of ordinary skill in the art will recognize that a 4-membered "heterocycloalkyl" or "heterocycle" may generally contain one or two heteroatoms, a 5- or 6-membered "heterocycloalkyl" or "heterocycle" may generally contain one, two, or three heteroatoms, and a 7- to 10-membered heterocycloalkyl or "heterocycle" may generally contain one, two, three, or four heteroatoms.

[0108]

[0074] "Heterocycloalkylene" refers to a divalent heterocycloalkyl as defined herein.

[0109] The term "heteroaryl" refers to a monovalent monocyclic aromatic group and / or polycyclic aromatic group in which at least one aromatic ring contains one or more heteroatoms independently selected from oxygen, sulfur, and nitrogen within the ring. Each ring of the heteroaryl group can contain one or two oxygen atoms, one or two sulfur atoms, and / or one to four nitrogen atoms, provided that the total number of heteroatoms within each ring is four or less and each ring contains at least one carbon atom. In certain embodiments, heteroaryl has 5 to 20, 5 to 15, or 5 to 10 ring atoms. Heteroaryl can be attached to the rest of the molecule via a nitrogen or carbon atom. In some embodiments, monocyclic heteroaryl groups include, but are not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, triazolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, and triazinyl. Examples of bicyclic heteroaryl groups include, but are not limited to, benzofuranyl, benzimidazolyl, benzoisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzotriazolyl, benzoxazolyl, furopyridyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quinazolinyl, thiadiazolopyrimidyl, and thienopyridyl. Examples of tricyclic heteroaryl groups include, but are not limited to, acridinyl, benzoindolyl, carbazolyl, dibenzofuranyl, perimidinyl, phenanthrolinyl, phenanthridinyl, phenarsazinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and xanthenyl. In certain embodiments, heteroaryl may also be optionally substituted as described herein."Replaced heteroaryl" is heteroaryl substituted as defined for aryl.

[0110]

[0076] The term "heteroarylene" as used herein refers to a divalent heteroaryl group as defined herein. "Replaced heteroarylene" is heteroarylene substituted as defined for aryl.

[0111]

[0077] The term "protecting group" as used herein, unless otherwise specified, refers to a group added to an oxygen, nitrogen, or phosphorus atom to prevent further reaction at (protected) oxygen, nitrogen, or phosphorus, or for other purposes. A wide variety of oxygen and nitrogen protecting groups are known to those of ordinary skill in the art of organic synthesis (see, e.g., Greene et al., Protective Groups in Organic Synthesis, John Wiley and Sons, 4th Edition, 2006, which is hereby incorporated by reference in its entirety).

[0112] "Pharmaceutically acceptable salts" refers to any salts of the compounds provided herein that retain their biological properties, are not toxic, and are not otherwise undesirable for pharmaceutical use. Such salts can be derived from a variety of organic and inorganic counterions well known in the art.Such salts include, but are not limited to, (1) acid addition salts formed by organic or inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, sulfamic acid, acetic acid, trifluoroacetic acid, trichloroacetic acid, propionic acid, hexanoic acid, cyclopentylpropionic acid, glycolic acid, glutaric acid, pyruvic acid, lactic acid, malonic acid, succinic acid, sorbic acid, ascorbic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, picric acid, caffeic acid, mandelic acid, phthalic acid, lauric acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphoric acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, cyclohexylsulfamic acid, quinic acid, and muconic acid; or (2) salts formed when the acidic proton present in the parent compound is replaced by (a) metal ions such as alkali metal ions, alkaline earth metal ions, or aluminum ions, or alkali metal hydroxides or alkaline earth metal hydroxides such as sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, aluminum hydroxide, lithium hydroxide, zinc hydroxide, and barium hydroxide, or ammonia, or (b) coordinated with organic bases such as aliphatic, alicyclic, or aromatic organic amines including, but not limited to, ammonia, methylamine, dimethylamine, diethylamine, picoline, ethanolamine, diethanolamine, triethanolamine, ethylenediamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylene-diamine, chloroprocaine, procaine, N-benzylphenethylamine, N-methylglucamine, piperazine, tris(hydroxymethyl)aminomethane, and tetramethylammonium hydroxide.

[0113]

[0079] Pharmaceutically acceptable salts further include, by way of example and not limitation, sodium salts, potassium salts, calcium salts, magnesium salts, ammonium salts, and tetraalkylammonium salts, etc. And when the compound contains a basic functional group, salts of hydrohalic acids, for example, hydrochloride salts and hydrobromide salts, sulfate salts, phosphate salts, sulfamate salts, nitrate salts, acetate salts, trifluoroacetate salts, trichloroacetate salts, propionate salts, hexanoate salts, cyclopentylpropionate salts, glycolate salts, glutarate salts, pyruvate salts, lactate salts, malonate salts, succinate salts, sorbate salts, ascorbate salts, malate salts, maleate salts, fumarate salts, tartrate salts, citrate salts, benzoate salts, 3-(4-hydroxybenzoyl)benzoate salts, picrate salts, caffeate salts, mandelate salts, phthalate salts, laurate salts, methanesulfonate salts (mesylate salts), ethanesulfonate salts, 1,2-ethane-disulfonate salts, 2-hydroxyethanesulfonate salts, benzenesulfonate salts (besylate salts), 4-chlorobenzenesulfonate salts, 2-naphthalenesulfonate salts, 4-toluenesulfonate salts, camphorate salts, camphorsulfonate salts, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylate salts, glucoheptonate salts, 3-phenylpropionate salts, trimethylacetate salts, tert-butylacetate salts, lauryl sulfate salts, gluconate salts, glutamate salts, hydroxynaphthoate salts, salicylate salts, stearate salts, cyclohexylsulfamate salts, quinate salts, muconate salts, etc., i.e., salts of non-toxic organic or inorganic acids are included.

[0114]

[0080] The terms "substantially free of" or "in the substantial absence of" with respect to a composition refer to a composition containing at least 85 wt%, 90 wt%, in certain embodiments, 95 wt%, 98 wt%, 99 wt%, or 100 wt%; or in certain embodiments, 95%, 98%, 99%, or 100% of the designated enantiomer or diastereomer of a compound. In certain embodiments, in the methods and compounds provided herein, the compound is substantially free of one of two enantiomers. In certain embodiments, in the methods and compounds provided herein, the compound is substantially free of one of two diastereomers. In certain embodiments, in the methods and compounds provided herein, the compound is substantially free of enantiomers (i.e., the compound is not a racemic or 50:50 mixture of the compound).

[0115]

[0081] Similarly, the term "isolated" with respect to a composition refers to a composition containing at least 85 wt%, 90 wt%, 95 wt%, 98 wt%, or 99 wt% to 100 wt% of a compound, with the remainder containing other chemical species, enantiomers, or diastereomers.

[0116]

[0082] "Solvate" refers to a compound or a salt thereof provided herein that further includes a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. When the solvent is water, the solvate is a hydrate.

[0117]

[0083] The term "substituted" refers to a moiety having a substituent that replaces one or more carbons or replaceable heteroatoms, such as hydrogen on NH or NH2, of a compound. It is understood that "substituted" or "substituted with" includes the implicit condition that such substitution follows the valences of the substituted atoms and the allowable atoms of the substituent and results in a stable compound, i.e., a compound that does not spontaneously undergo transformation, such as rearrangement, cyclization, elimination, etc. In certain embodiments, "substituted" refers to a moiety having a substituent that replaces two hydrogen atoms on the same carbon atom, such as replacing two hydrogen atoms on a single carbon with an oxo, imino, or thioxo group. As used herein, the term "substituted" is intended to include all acceptable substituents of an organic compound. In a broad aspect, acceptable substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of an organic compound. Acceptable substituents may be one or more, and may be the same or different, for a suitable organic compound.

[0118]

[0084] In some embodiments, the substituent is any of the substituents described herein, such as: halogen, hydroxy, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO2), imino (=N-H), oximo (=N-OH), hydrazino (=N-NH2), -R b1 -OR a1 、-R b1 -OC(O)-R a1 、-R b1 -OC(O)-OR a1 、-R b1 -OC(O)-N(R a1 )2、-R b1 -N(R a )2、-R b1 -C(O)R a1 、-R b1 -C(O)OR a1 、-R b1 -C(O)N(R a1 )2、-R b1 -O-R c1 -C(O)N(R a1 )2、-Rb1 -N(R a1 )C(O)OR a1 、-R b1 -N(R a )C(O)R a1 、-R b1 -N(R a1 )S(O) t R a1 (where t is 1 or 2), -R b1 -S(O) t R a1 (where t is 1 or 2), -R b1 -S(O) t OR a1 (where t is 1 or 2), and -R b1 -S(O) t N(R a1 )2(where t is 1 or 2); and any of them being alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo(=O), thioxo(=S), cyano(-CN), nitro(-NO2), imino(=N-H), oximo(=N-OH), hydrazine(=N-NH2), -R b1 -OR a1 、-R b1 -OC(O)-R a1 、-R b1 -OC(O)-OR a1 、-R b1 -OC(O)-N(R a1 )2、-R b1 -N(R a )2、-R b1 -C(O)R a1 、-R b1 -C(O)OR a1 、R b1 -C(O)N(R a1 )2、-R b1 -O-R c1 -C(O)N(R a1 )2、-R b1 -N(R a1 )C(O)OR a1 、-R b1 -N(R a1 )C(O)R a1 、-R b1 -N(R a1 )S(O)t R a1 (wherein t is 1 or 2), -R b1 -S(O) t R a1 (wherein t is 1 or 2), -R b1 -S(O) t OR a1 (wherein t is 1 or 2) and -R b1 -S(O) t N(R a1 )2 (wherein t is 1 or 2), alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl which may be optionally substituted by -R a1 each R a1 is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, and each R b1 -OR a1 -R b1 -OC(O)-R a1 -R b1 -OC(O)-OR a1 -R b1 -OC(O)-N(R a1 )2, -R b1 -N(R a1 )2, -R b1 -C(O)R a1 -R b1 -C(O)OR a1 -R b1 -C(O)N(R a1 )2, -R b1 -O-R c1 -C(O)N(R a1 )2, -R b1 -N(Ra1 )C(O)OR a1 、 -R b1 -N(R a1 )C(O)R a1 、 -R b1 -N(R a )S(O) t R a1 (wherein t is 1 or 2), -R b1 -S(O) t R a1 (wherein t is 1 or 2), -R b1 -S(O) t OR a1 (wherein t is 1 or 2) and -R b1 -S(O) t N(R a1 )2 (wherein t is 1 or 2) may be optionally substituted; each R b1 is independently selected from a direct bond, or a straight or branched alkylene, alkenylene, or alkynylene chain, and each R c is a straight or branched alkylene, alkenylene, or alkynylene chain) may be included.

[0119]

[0085] It will be understood by those skilled in the art that, where appropriate, the substituents themselves may also be substituted. Unless otherwise specified as "unsubstituted", references to chemical moieties herein are understood to include substituted variants. For example, references to a "heteroaryl" group or moiety, unless otherwise specified, implicitly include both substituted and unsubstituted variants.

[0120]

[0086] The term "amino acid" or "amino acid residue" refers to D- or L-amino acids, whether natural or non-natural. Representative amino acids include, but are not limited to, alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan, or tyrosine. "Non-natural amino acids" are amino acids that do not form part of a protein and are either naturally occurring or chemically synthesized. Non-limiting examples of non-natural amino acids include sulfoalanine, hydroxyproline (Hyp), β-alanine, citrulline (Cit), ornithine (Orn), norleucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, and methionine sulfone.

[0121]

[0087] "Isotopic composition" refers to the amount of each isotope present for a given atom, and "natural isotopic composition" refers to the isotopic composition or abundance that is naturally present for a given atom. Atoms containing their natural isotopic composition may also be referred to herein as "non-enriched" atoms. Unless otherwise indicated, the atoms of the compounds recited herein are intended to represent any stable isotope of that atom. For example, unless otherwise explicitly stated, if a position is specifically shown as hydrogen (H), that position is understood to have hydrogen with its natural isotopic composition.

[0122]

[0088] "Isotope enrichment" refers to the proportion of incorporation of a specific isotope of an atom into a molecule, instead of the natural isotope abundance of a given atom. For example, 1% deuterium (D) enrichment at a given position means that 1% of the molecules in a given sample contain deuterium at the designated position. Since the naturally occurring deuterium distribution is about 0.0156%, the deuterium enrichment at any position in a compound synthesized using non-enriched starting materials is about 0.0156%. The isotope enrichment of the compounds provided herein can be determined using conventional analytical methods known to those skilled in the art, including mass spectrometry and nuclear magnetic resonance spectroscopy.

[0123]

[0089] "Isotope-enriched" refers to an atom having an isotope composition other than the natural isotope composition of the atom. "Isotope-enriched" can also refer to a compound containing at least one atom having an isotope composition other than the natural isotope composition of the atom.

[0124]

[0090] As used herein, the terms "alkyl", "alkylene", "alkylamino", "dialkylamino", "cycloalkyl", "aryl", "arylene", "alkoxy", "amino", "carboxyl", "heterocycloalkyl", "heteroaryl", "heteroarylene", "carboxyl", and "amino acid" groups optionally include deuterium (D) at one or more positions where a hydrogen (H) atom is present, and the deuterium composition of one or more atoms is other than the natural isotope composition.

[0125]

[0091] Similarly, as used herein, the terms "alkyl", "alkylene", "alkylamino", "dialkylamino", "cycloalkyl", "aryl", "arylene", "alkoxy", "amino", "carboxyl", "heterocycloalkyl", "heteroaryl", "heteroarylene", "carboxyl", and "amino acid" groups optionally include an amount of carbon-13 ( 13 C) other than the natural isotope composition.

[0126] As used herein, the term "polymer" or "polymeric moiety" refers to proteins, peptides, antibodies, nucleic acids, carbohydrates, or other large molecules composed of polymerized monomers. This includes peptides having two or more residues, or ten or more residues. In certain embodiments, the polymer has a mass of at least 1000 Da. In certain embodiments, the polymer has at least 1000 atoms. In certain embodiments, the polymer can be modified. For example, a protein, peptide, or antibody can be modified with one or more carbohydrates and / or small molecule therapeutic compounds.

[0127]

[0093] As used herein, the term "immunoglobulin" refers to a class of structurally related proteins generally comprising two pairs of polypeptide chains, one pair of light (L) chains and one pair of heavy (H) chains. In "intact immunoglobulins", all four of these chains are interconnected by disulfide bonds. The structure of immunoglobulins is well-characterized. See, e.g., Paul, Fundamental Immunology, 7th Edition, Chapter 5 (2013) Lippincott Williams & Wilkins, Philadelphia, PA. Briefly, each heavy chain typically includes a heavy chain variable region (V H or VH) and a heavy chain constant region (C H or CH). The heavy chain constant region typically includes three domains designated C H 1 (or CH1), C H 2 (or CH2), and C H 3 (or CH3). Each light chain typically includes a light chain variable region (V L or VL) and a light chain constant region. The light chain constant region typically includes one domain designated C L or CL.

[0128] The term "antibody" is used herein in its broadest sense. Antibodies include intact antibodies (e.g., intact immunoglobulins), and antibody fragments (e.g., antigen-binding fragments or antigen-binding fragments of antibodies). An antibody includes at least one antigen-binding domain. An example of an antigen-binding domain is a V H -V L antigen-binding domain formed by a dimer.

[0129] The term "amino acid" refers to the 20 common naturally occurring amino acids. Naturally occurring amino acids include alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine (Cys; C), glutamic acid (Glu; E), glutamine (Gln; Q), glycine (Gly; G), histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Val; V), as well as the less common pyrrolidine and selenocysteine. Naturally occurring amino acids also include citrulline. Naturally encoded amino acids include post-translational variants of 22 naturally occurring amino acids such as prenylated amino acids, isoprenylated amino acids, myristoylated amino acids, palmitoylated amino acids, N-linked glycosylated amino acids, O-linked glycosylated amino acids, phosphorylated amino acids, and acylated amino acids. The term "amino acid" includes, but is not limited to, non-natural (or unnatural) or synthetic α-, β-, γ-, or δ-amino acids, including the amino acids found in proteins, namely glycine, alanine, valine, leucine, isoleucine, methionine, phenylalanine, tryptophan, proline, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, and histidine. In certain embodiments, the amino acid is in the L configuration. In certain embodiments, the amino acid is in the D configuration.Alternatively, the amino acid can be a derivative of alanyl, valinyl, leucinyl, isoleucinyl, prolynyl, phenylalanyl, tryptophanyl, methioninyl, glycyl, serynyl, threoninyl, cysteinyl, tyrosinyl, asparaginyl, glutaminyl, aspartoyl, glutaryl, lysinyl, argininyl, histidinyl, β-alanyl, β-valinyl, β-leucinyl, β-isoleucinyl, β-prolynyl, β-phenylalanyl, β-tryptophanyl, β-methioninyl, β-glycyl, β-serynyl, β-threoninyl, β-cysteinyl, β-tyrosinyl, β-asparaginyl, β-glutaminyl, β-aspartoyl, β-glutaryl, β-lysinyl, β-argininyl, or β-histidinyl. A non-natural amino acid is neither an amino acid that constitutes a protein nor a post-translational modification variant thereof. In particular, the term non-natural amino acid refers to an amino acid that is not one of the 20 common amino acids, nor pyrrolidine, nor selenocysteine, nor a post-translational modification variant thereof.

[0130]

[0096] The term "conjugate" refers to a compound or drug moiety described herein that is linked to one or more polymeric moieties. The polymeric moiety is as defined herein or any polymer that is considered suitable for one of ordinary skill in the art. The compound or drug moiety can be any compound or drug moiety described herein. The compound or drug moiety can be directly linked to the polymeric moiety via a covalent bond, or the compound or drug moiety can be indirectly linked to the polymeric moiety via a linker. Typically, the linker is covalently bonded to the polymeric moiety and also covalently bonded to the compound or drug moiety.

[0131]

[0097] "pAMF", "pAMF residue", or "pAMF variant" refers to a mutant phenylalanine residue (i.e., para-azidomethyl-L-phenylalanine) added or substituted to a polypeptide.

[0132] As used herein, the term "linker" refers to a molecular moiety capable of forming at least two covalent bonds. Typically, a linker is capable of forming at least one covalent bond with a polymeric moiety and at least one additional covalent bond with a compound or drug moiety. In certain embodiments, a linker is capable of forming more than two covalent bonds with a polymeric moiety. In certain embodiments, a linker is capable of forming more than two covalent bonds with a compound or drug moiety, or is capable of forming covalent bonds with more than two compound or drug moieties. After the linker has formed a bond with a polymeric moiety, or a compound or drug moiety, or both, the remaining structure (i.e., the "linker residue" of the linker after one or more covalent bonds have been formed) may still be referred to herein as the "linker". The term "linker precursor" refers to a linker having one or more reactive groups capable of forming a covalent bond with a polymer, or a compound or drug moiety, or both. One of ordinary skill in the art will understand, in view of the context in which the term "linker" is used, whether "linker" means a linker precursor having one reactive group, a linker precursor having more than one reactive group, a linker residue covalently attached to a polymer, a linker residue covalently attached to a compound or drug moiety, and / or a linker residue covalently attached to a polymer and covalently attached to a compound or drug moiety. In some embodiments, the linker is a cleavable linker. For example, a cleavable linker may be labile or released by an enzymatic function, whether or not it has been manipulated. In some embodiments, the linker is a non-cleavable linker. For example, a non-cleavable linker may be released by degradation of the polymeric moiety.

[0133]

[0099] As used herein, "EC 50 " refers to the dose, concentration, or amount of a particular test compound that elicits a dose-dependent response at 50% of the maximum manifestation of a particular response induced, evoked, or enhanced by the particular test compound.

[0134]

[0100] As used herein, unless otherwise specified, the term "IC" 50 refers to the amount, concentration, or dosage of a particular test compound that achieves 50% inhibition of the maximal response in an assay measuring a response.

[0135]

[0101] As used herein, the terms "subject" and "patient" are used interchangeably. The terms "subject" and "subjects" refer to animals such as mammals including non - primates (e.g., cows, pigs, horses, cats, dogs, rats, and mice) and primates (e.g., monkeys such as cynomolgus monkeys, chimpanzees, and humans), and in certain embodiments, refer to humans. In certain embodiments, the subject is a farm animal (e.g., horse, cow, pig, etc.) or a pet (e.g., dog or cat). In certain embodiments, the subject is a human.

[0136]

[0102] As used herein, the terms "therapeutic agent" and "therapeutic agents" refer to any agent that can be used for the treatment or prevention of a disorder or one or more of its symptoms. In certain embodiments, the term "therapeutic agent" includes the compounds or conjugates provided herein. In certain embodiments, the therapeutic agent is an agent that is known to be useful or has been used or is currently used for the treatment or prevention of a disorder or one or more of its symptoms.

[0137]

[0103] "Therapeutically effective amount" refers to the amount of a compound or composition that, when administered to a subject for treating a condition, is sufficient to effect treatment of such condition. The "therapeutically effective amount" can vary particularly depending on the compound, the disease or disorder and its severity, and the age, weight, etc. of the subject being treated.

[0138]

[0104] "Treating" or "treatment" of any disease or disorder, in certain embodiments, refers to ameliorating the disease or disorder present in a subject. In another embodiment, "treating" or "treatment" includes improving at least one physical parameter that may be undetectable by the subject. In yet another embodiment, "treating" or "treatment" includes modulating the disease or disorder, either physically (e.g., stabilization of discernible symptoms) or physiologically (e.g., stabilization of physical parameters) or both. In yet another embodiment, "treating" or "treatment" includes delaying or preventing the onset of a disease or disorder, or delaying or preventing the recurrence of a disease or disorder. In yet another embodiment, "treating" or "treatment" includes reducing or eliminating any of a disease or disorder, or delaying the progression of a disease or disorder or one or more symptoms thereof, or reducing the severity of a disease or disorder or one or more symptoms thereof.

[0139]

[0105] As used herein, the term "inhibiting growth" (e.g., with respect to cells such as tumor cells) is intended to include any measurable decrease in cell growth (e.g., tumor cell growth) as compared to the growth of the same cells not in contact with a compound, drug moiety, or conjugate of the present disclosure when contacted with a compound, drug moiety, or conjugate of the present disclosure. In some embodiments, growth can be inhibited by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 99%, or 100%. The decrease in cell growth can occur via various mechanisms including, but not limited to, internalization, apoptosis, necrosis, and / or effector function-mediated activity of the conjugate, compound, or drug moiety.

[0140] As used herein, the terms "prophylactic agent" and "prophylactic agents" refer to any agent that can be used to prevent a disorder or one or more symptoms thereof. In certain embodiments, the term "prophylactic agent" includes the compounds, drug moieties, or conjugates provided herein. In certain other embodiments, the term "prophylactic agent" does not refer to any of the compounds, drug moieties, or conjugates provided herein. For example, a prophylactic agent is known to be useful for preventing or interfering with the onset, development, progression, and / or severity of a disorder, or has been used or is currently being used to prevent or interfere with the onset, development, progression, and / or severity of a disorder.

[0141] As used herein, the phrase "prophylactically effective amount" refers to an amount of a treatment (e.g., a prophylactic agent) sufficient to effect prevention or reduction of the development, recurrence, or onset of one or more symptoms associated with a disorder, or to enhance or improve the prophylactic effect of another treatment (e.g., another prophylactic agent).

[0142] In some of the chemical structures exemplified herein, certain substituents, chemical groups, and atoms are depicted by one or more curved / wavy / sinuous lines that cross one or more bonds to indicate the atom to which the substituent, chemical group, and atom are attached (e.g.,

[0143]

Chemical Structure

[0144]

Chemical Structure

[0145] [Chem.] In some structures, such as, this curved / wavy / sinuous line indicates atoms in a polymer, as well as atoms in the backbone of a conjugate, compound, or drug moiety to which the exemplified chemical entity is attached.

[0146]

[0109] As used herein, a figure showing a substituent attached to a cyclic group (e.g., aromatic, heteroaromatic, fused ring, and saturated or unsaturated cycloalkyl or heterocycloalkyl) through a bond between ring atoms, unless otherwise specified, is intended to show that the cyclic group may be substituted with that substituent at any ring position of the cyclic group or at any ring of a fused ring group, as shown herein or according to techniques known in the art to which this disclosure pertains. For example, a group,

[0147] [Chem.] (wherein the position of the substituent O-Su is generally described, i.e., at any vertex of the bond-line structure, i.e., not directly attached to a specific ring carbon atom) is the following non-limiting examples of groups in which the substituent O-Su is attached to a specific ring atom:

[0148] [Chem.] including.

[0149] As used herein, the term "site-specific" refers to the modification of a polypeptide at a defined sequence position of the polypeptide. The modification is at a single, predictable residue of the polypeptide with little or no variation. In certain embodiments, the modified amino acid is introduced recombinantly or synthetically at that sequence position. Similarly, a moiety can be "site-specifically" linked to a residue at a defined sequence position of a polypeptide. In certain embodiments, a polypeptide can include two or more site-specific modifications.

[0150]

[0111] Compounds of formulas (I) and (III) and conjugates of formulas (II) and (IV) In one aspect, the compound is a compound of formula (I):

[0151]

Chemical formula

[0152]

Chemical formula

[0153]

Chemical formula

[0154]

Chemical formula

[0155]

[0112] In certain embodiments, the compound of formula (I) is a compound of formula (IA):

[0156]

Chemical formula

[0157]

[0113] In certain embodiments, the compound of formula (IA) is selected from the following:

[0158]

Chemical formula

[0159]

[0114] In certain embodiments, the compound of formula (I) is a compound of formula (IB):

[0160]

Chemical formula

[0161]

[0115] In certain embodiments, the compound of formula (IB) is selected from the following:

[0162]

Chemical formula

[0163]

[0116] In certain embodiments, the compound of formula (I) is a compound of formula (IC):

[0164]

Chemical formula

[0165]

[0117] In certain embodiments, the compound of formula (IC) is selected from the following:

[0166]

Chemical formula

[0167]

[0118] In certain embodiments, the compound of formula (I) is a compound of formula (ID):

[0168]

Chemical formula

[0169]

[0119] In certain embodiments, the compound of formula (ID) is one of the following:

[0170]

Chemical formula

[0171]

[0120] In certain embodiments, the compound of formula (I) is a compound of formula (IE):

[0172]

Chemical formula

[0173]

[0121] In certain embodiments, the compound of formula (IE) is one of the following:

[0174]

Chemical formula

[0175]

Chem.

[0176]

[0122] In certain embodiments, the compound of formula (I) is a compound of formula (IF):

[0177]

Chem.

[0178]

[0123] In certain embodiments, the compound of formula (IF) is as follows:

[0179]

Chem.

[0180]

[0124] In certain embodiments, the compound of formula (I) is a compound of formula (IG):

[0181]

Chem.

[0182]

[0125] In certain embodiments, the compound of formula (IG) is as follows:

[0183]

Chemical formula

[0184]

[0126] In certain embodiments, the compound of formula (I) is a compound of formula (IH):

[0185]

Chemical formula

[0186]

[0127] In certain embodiments, the compound of formula (IH) is as follows:

[0187]

Chemical formula

[0188]

[0128] In one aspect, the conjugate of formula (II):

[0189]

Chemical formula

[0190]

Chemical formula

[0191]

Chem.

[0192]

Chem.

[0193]

[0129] In certain embodiments, the compound of formula (II) is a compound of formula (IIA):

[0194]

Chem.

[0195]

[0130] In certain embodiments, the compound of formula (IA) is selected from the following:

[0196]

Chemical formula

[0197]

[0131] In certain embodiments, the compound of formula (II) is the compound of formula (IIB):

[0198]

Chemical formula

[0199]

[0132] In certain embodiments, the compound of formula (IIB) is selected from the following:

[0200]

Chemical formula

[0201]

[0133] In certain embodiments, the compound of formula (II) is a compound of formula (IIC):

[0202]

Chemical formula

[0203]

[0134] In certain embodiments, the compound of formula (IIC) is one of the following:

[0204]

Chemical formula

[0205]

[0135] In certain embodiments, the compound of formula (II) is a compound of formula (IID):

[0206]

Chemical formula

[0207]

[0136] In certain embodiments, the compound of formula (IID) is one of the following:

[0208] [Chemical formula] Or selected from its pharmaceutically acceptable salts and / or stereoisomers.

[0209]

[0137] In certain embodiments, the compound of formula (II) is a compound of formula (IIE):

[0210] [Chemical formula] Or its pharmaceutically acceptable salts and / or stereoisomers (wherein the integer a, integer c, RL, R a , R b , ring B, L 3 , POLY 2 , D, and COMP are as defined herein) .

[0211]

[0138] In certain embodiments, the compound of formula (IIE) is the following:

[0212] [Chemical formula]

[0213] [Chemical formula] Or selected from its pharmaceutically acceptable salts and / or stereoisomers.

[0214]

[0139] In certain embodiments, the compound of formula (II) is a compound of formula (IIF):

[0215] [Chemical formula] or a pharmaceutically acceptable salt and / or stereoisomer thereof (wherein integer a, integer c, RL, R a 、R b 、ring B, L 3 、POLY 2 、D, and COMP are as defined herein) is as follows.

[0216]

[0140] In certain embodiments, the compound of formula (IIF) is selected from the following:

[0217]

Chemical Structure

[0218]

[0141] In certain embodiments, the compound of formula (II) is the compound of formula (IIG):

[0219]

Chemical Structure

[0220]

[0142] In certain embodiments, the compound of formula (IIG) is selected from the following:

[0221]

Chemical Structure

[0222]

[0143] In certain embodiments, the compound of formula (II) is a compound of formula (IIH):

[0223]

Chemical formula

[0224]

[0144] In certain embodiments, the compound of formula (IIH) is one of the following:

[0225]

Chemical formula

[0226]

[0145] In one aspect, a conjugate of formula (III):

[0227]

Chemical formula

[0228]

[0146] Non-limiting non-natural amino acids include sulfonalanine, hydroxyproline (Hyp), β-alanine, citrulline (Cit), ornithine (Orn), norleucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, and methionine sulfone.

[0229]

[0147] In certain embodiments, the compound of formula (III) is a compound of formula (IIIA):

[0230]

Chem.

[0231]

[0148] In certain embodiments, the compound of formula (III) is a compound of formula (IIIB):

[0232]

Chem.

[0233]

[0149] In certain embodiments, the compound of formula (IIIB) is of the formula:

[0234]

Chem.

[0235]

[0150] In one embodiment, a conjugate of formula (IV):

[0236]

Chemical Formula

[0237]

[0151] In certain embodiments, a compound of formula (IV) is a compound of formula (IVA):

[0238]

Chemical Formula

[0239]

[0152] In certain embodiments, a compound of formula (IV) is a compound of formula (IVB):

[0240]

Chemical Formula

[0241]

[0153] In certain embodiments, the compound of formula (IVB) is of the formula:

[0242]

Chem.

[0243]

[0154] In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 is

[0244]

Chem.

[0245]

Chem.

[0246]

Chem.

[0247]

Chem.

[0248]

Chem.

[0249]

Chem.

[0250]

Chem.

[0251]

Chem.

[0252]

Chem.

[0253]

Chem.

[0254] [Chemistry] is. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, L 1 is

[0255] [Chemistry] is. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, L 1 is

[0256] [Chemistry] is.

[0257]

[0155] In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, L 1 is

[0258] [Chemistry] is. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, L 1 is

[0259] [Chemistry] is.

[0260]

[0156] In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, ring B of L 1 is a 5- to 12-membered N-linked bridged, fused, or spiro bicyclic heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, including the N to which the ring is attached. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, L 1Ring B is a 5- to 12-membered N-bonded spirobicyclic heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, including the N to which the ring is bonded.

[0261]

[0157] In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 Ring B is an optionally substituted 5- to 12-membered N-bonded, bridged, fused, or spirobicyclic heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, including the N to which the ring is bonded, and the heterocyclic ring of Ring B is C 1~12 alkyl, C 2~12 alkenyl, C 2~12 alkynyl, C 3~12 cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)-, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl, and is optionally substituted with one or more substituents selected from the foregoing. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 Ring B is an optionally substituted 5- to 12-membered N-bonded spirobicyclic heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, including the N to which the ring is bonded, and the heterocyclic ring of Ring B is C 1~12 alkyl, C 2~12 alkenyl, C 2~12 alkynyl, C 3~12 cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)-, -C(O)N(R 2 R 3 )2, -C(O)OR 2It may be substituted with one or more substituents selected from aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

[0262]

[0158] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, L 1 the ring B of

[0263]

Chemical formula

[0264]

[0159] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, L 1 the ring B of

[0265]

Chemical formula

[0266]

Chemical formula

[0267]

Chemical formula

[0268]

Chemical formula

[0269]

Chem.

[0270]

Chem.

[0271]

Chem.

[0272]

[0160] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, L 1 ring B of

[0273]

Chem.

[0274]

Chem.

[0275]

Chem.

[0276]

Chem.

[0277]

Chem.

[0278]

Chem.

[0279]

Chem.

[0280]

[0161] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, L 1 ring B of

[0281]

Chem.

[0282]

Chem.

[0283]

Chem.

[0284]

Chem.

[0285]

Chem.

[0286]

Chem.

[0287]

Chem.

[0288]

Chem.

[0289]

Chem.

[0290]

[0162] In certain embodiments of formulas (I) - (IIH),

[0291]

Chem.

[0292]

Chem.

[0293]

Chem.

[0294] [Chem.] as follows. In certain embodiments of formulas (I)-(IIH),

[0295] [Chem.] is

[0296] [Chem.] as follows. In certain embodiments of formulas (I)-(IIH),

[0297] [Chem.] is

[0298] [Chem.] as follows. In certain embodiments of formulas (I)-(IIH),

[0299] [Chem.] is

[0300] [Chem.] as follows.

[0301]

[0163] In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 ring B of

[0302] [Chemical formula] is. In certain embodiments of formulas (I) to (IIH),

[0303] [Chemical formula] where

[0304] [Chemical formula] is, and m is 1, 2, or 3. In certain embodiments of formulas (I) to (IIH),

[0305] [Chemical formula] where

[0306] [Chemical formula] is, and m is 1, 2, or 3. In certain embodiments of formulas (I) to (IIH),

[0307] [Chemical formula] where

[0308] [Chemical formula] is, and m is 1, 2, or 3. In certain embodiments of formulas (I) to (IIH),

[0309] [Chemical formula] where

[0310]

Chem.

[0311]

[0164] In certain embodiments of formulas (I) - (IIH),

[0312]

Chem.

[0313]

Chem.

[0314]

Chem.

[0315]

Chem.

[0316]

Chem.

[0317]

Chem.

[0318]

Chem.

[0319]

Chem.

[0320]

Chem.

[0321]

Chem.

[0322]

[0165] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, L 1 ring B of is

[0323]

Chem.

[0324]

[0166] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, L 1 ring B of is

[0325]

Chem.

[0326]

[0167] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, L 1 ring B of is

[0327]

Chem.

[0328]

[0168] In certain embodiments of formulas (I)-(IIH) that include any of the foregoing, L 1 is

[0329]

Chem.

[0330]

[0169] In certain embodiments of formulas (I)-(IIH) that include any of the foregoing, L 1 is

[0331]

Chem.

[0332]

[0170] In certain embodiments of formulas (I)-(IIH) that include any of the foregoing, ring B of L 1 is

[0333]

Chem.

[0334]

[0171] In certain embodiments of formulas (I)-(IIH) including any of the foregoing, ring B of L 1 is

[0335]

Chemical formula

[0336]

[0172] In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L1 The ring A of [description] is a bridged, fused, or spiro bicyclic carbocyclic ring. In certain embodiments including any of the foregoing, L 1 the ring A of [description] is C 4~12 a bridged, fused, or spiro bicyclic carbocyclic ring. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 the ring A of [description] is C 4~12 a bridged bicyclic carbocyclic ring. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 the ring A of [description] is C 4~8 a bridged bicyclic carbocyclic ring.

[0337]

[0173] In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 the ring A of [description] is an optionally substituted bridged, fused, or spiro bicyclic carbocyclic ring, and the carbocyclic ring of ring A is C 1~12 alkyl, C 2~12 alkenyl, C 2~12 alkynyl, C 3~12 cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)-, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl, and may be substituted with one or more substituents selected from the group consisting of: In certain embodiments including any of the foregoing, L 1 the ring A of [description] is an optionally substituted C 4~12 bridged, fused, or spiro bicyclic carbocyclic ring. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 the ring A of [description] is an optionally substituted C 4~12 bridged bicyclic carbocyclic ring. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, L 1 the ring A of [description] is an optionally substituted C 4~8 bridged bicyclic carbocyclic ring.

[0338]

[0174] In certain embodiments of formulas (I)-(IIH) comprising any of the foregoing, L 1 the ring A of

[0339]

Chemical formula

[0340]

[0175] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, ring A of L 1 is selected from

[0341]

Chemical formula

[0342]

[0176] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, ring A of L 1 is selected from

[0343]

Chemical formula

[0344]

[0177] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, ring A of L 1 is selected from

[0345]

Chemical formula

[0346]

[0178] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, ring A of L 1 is

[0347]

Chem.

[0348]

[0179] In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, ring A of L 1 is

[0349]

Chem.

[0350]

Chem.

[0351]

[0180] In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, X 1 , X 2 , X 3 , and / or X 4 is -C(R 4 )2-. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, X 1 and X 2 are -C(R 4 )2-. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, X 1 , X 2 , and X 3 are -C(R 4 )2-. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, X 1 , X 2 , X 3 , and X 4 are -C(R 4) is 2-. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, X 1 is -NH-. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, X 2 is -NH-. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, X 3 is -NH-. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, X 4 is -NH-. In certain embodiments including any of the foregoing, X 1 is -O-. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, X 2 is -O-. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, X 3 is -O-. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, X 4 is -O-.

[0352]

[0181] In certain embodiments of formulas (I) to (IIH) including any of the foregoing, L 1 is

[0353]

Chem.

[0354]

Chem.

[0355]

[0182] In certain embodiments including any of the foregoing, a is 0. In certain embodiments including any of the foregoing, a is 1. In certain embodiments including any of the foregoing, a is 2. In certain embodiments including any of the foregoing, a is 3. In certain embodiments including any of the foregoing, a is 4. In certain embodiments including any of the foregoing, a is 5. In certain embodiments including any of the foregoing, a is 6.

[0356]

[0183] In certain embodiments of formulas (I) to (IIH), b is 0. In certain embodiments of formulas (I) to (IIH), b is 1.

[0357]

[0184] In certain embodiments of formulas (I) to (IIH), b is 0 and a is 0. In certain embodiments of formulas (I) to (IIH), b is 0 and a is 1. In certain embodiments of formulas (I) to (IIH), b is 0 and a is 2. In certain embodiments of formulas (I) to (IIH), b is 0 and a is 3. In certain embodiments of formulas (I) to (IIH), b is 0 and a is 4. In certain embodiments of formulas (I) to (IIH), b is 0 and a is 5. In certain embodiments of formulas (I) to (IIH), b is 0 and a is 6. In certain embodiments of formulas (I) to (IIH), b is 1 and a is 1. In certain embodiments of formulas (I) to (IIH), b is 1 and a is 2. In certain embodiments of formulas (I) to (IIH), b is 1 and a is 3. In certain embodiments of formulas (I) to (IIH), b is 1 and a is 4. In certain embodiments of formulas (I) to (IIH), b is 1 and a is 5. In certain embodiments of formulas (I) to (IIH), b is 1 and a is 6.

[0358]

[0185] In certain embodiments of formulas (I) to (IIH) including any of the foregoing, R 1 is hydrogen. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, R 1is unsubstituted alkyl. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, R 1 is methyl. In certain embodiments of formulas (I) to (IIH) including any of the foregoing, R 1 is alkyl optionally substituted with one or more substituents selected from cycloalkyl, halogen, alkoxy, -CN, -NO2, and -OH.

[0359]

[0186] In certain embodiments of formulas (III) to (IVB), L 5 is a linker comprising at least one amino acid selected from sulfonalanine, hydroxyproline (Hyp), β-alanine, citrulline (Cit), ornithine (Orn), norleucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, and methionine sulfone. In certain embodiments of formulas (III) to (IVB), L 5 is

[0360]

Chemical formula

[0361]

Chemical formula

[0362]

Chemical formula

[0363] [Chemical formula] is as follows.

[0364]

[0187] In certain embodiments including any of the foregoing, R a is hydrogen, and R b is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl. In certain embodiments including any of the foregoing, R a is hydrogen, and R b is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, -C(O)OH, aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl. In certain embodiments including any of the foregoing, R a is hydrogen, and R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH. In certain embodiments including any of the foregoing, R a and R b are both hydrogen.

[0365]

[0188] In certain embodiments of formulas (I) - (IIH) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; R 1is hydrogen; a is 1; b is 1. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; R 1 is hydrogen; a is 2; b is 1. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; R 1 is hydrogen; a is 3; b is 1.

[0366]

[0189] In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; a is 1; b is 0. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; a is 2; b is 0. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; a is 3; b is 0.

[0367]

[0190] In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a is hydrogen; R bis selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; R 1 is methyl; a is 1; b is 1. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; R 1 is methyl; a is 2; b is 1. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; R 1 is methyl; a is 3; b is 1.

[0368]

[0191] In certain embodiments including any of the foregoing of formulas (I)-(IIH), R a and R b are both hydrogen; R 1 is hydrogen; a is 1, and b is 1. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is hydrogen; a is 2, and b is 1. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is hydrogen; a is 3, and b is 1. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is hydrogen; a is 4, and b is 1. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R1 is hydrogen; a is 5 and b is 1. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is hydrogen; a is 6 and b is 1.

[0369]

[0192] In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; a is 1; b is 0. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; a is 2; b is 0. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; a is 3; b is 0. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; a is 4; b is 0. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; a is 5; b is 0. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; a is 6; b is 0.

[0370]

[0193] In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is methyl; a is 1; b is 0. In certain embodiments of Formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1is methyl; a is 2; b is 0. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is methyl; a is 3; b is 0. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is methyl; a is 4; b is 0. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is methyl; a is 5; b is 0. In certain embodiments of formulas (I)-(IIH) including any of the foregoing, R a and R b are both hydrogen; R 1 is methyl; a is 6; b is 0.

[0371]

[0194] In certain embodiments of formulas (III)-(IVB) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; a is 1; c is 1. In certain embodiments of formulas (III)-(IVB) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; a is 2; c is 1. In certain embodiments of formulas (III)-(IVB) including any of the foregoing, R a is hydrogen; R b is selected from hydrogen, alkyl, halogen, alkoxy, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)OH; a is 3; c is 1.

[0372]

[0195] In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b ) c -NH-, where * represents the location where Y is attached to RG in Formulas (I)-(IH) and (III)-(IIIB), and to RL in Formulas (II)-(IIH) and (IV)-(IVB). In certain embodiments, including any of the foregoing, Y is *-C(O)-(CH2) c -NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CH2)-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CH2)2-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CH2)3-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CH2)4-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CH2)5-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CH2)6-NH-.

[0373]

[0196] In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b )-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b )2-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b )3-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b )4-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b )5-NH-. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b )6-NH-.

[0374]

[0197] In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b ) c -NH-, R a is hydrogen, and R b is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl. In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b )2-NH-, *-C(O)-(CR a R b )3-NH-, or *-C(O)-(CR a R b )4-NH-, R a is hydrogen, and R b is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

[0375]

[0198] In certain embodiments, including any of the foregoing, Y is *-C(O)-(CR a R b ) c- and *, where Y is attached to RG in Formulas (I) to (IH) and (III) to (IIIB), and to RL in Formulas (II) to (IIH) and (IV) to (IVB). In certain embodiments including any of the foregoing, Y is *-C(O)-(CH2) c -. In certain embodiments including any of the foregoing, Y is *-C(O)-(CH2)-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CH2)2-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CH2)3-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CH2)4-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CH2)5-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CH2)6-.

[0376]

[0199] In certain embodiments including any of the foregoing, Y is *-C(O)-(CR a R b )-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CR a R b )2-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CR a R b )3-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CR a R b )4-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CR a R b )5-. In certain embodiments including any of the foregoing, Y is *-C(O)-(CR a R b )6-.

[0377]

[0200] In certain embodiments including any of the foregoing, Y is *-C(O)-(CR a R b ) c - and R a is hydrogen and Rb is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl. In certain embodiments, including any of the foregoing, Y is, *-C(O)-(CR a R b )2-, *-C(O)-(CR a R b )3-, or *-C(O)-(CR a R b )4-, R a is hydrogen, and R b is selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO2, -OH, -N(R 2 R 3 )2, -C(O)N(R 2 R 3 )2, -C(O)OR 2 , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

[0378]

[0201] In certain embodiments, including any of the foregoing, Y is *-C(O)-(CH2)2-NH- or *-C(O)-(CH2)4-.

[0379]

[0202] In certain embodiments, including any of the foregoing, L 2 is absent. In certain embodiments, including any of the foregoing, L 2 is a linker comprising a hydrophilic polymer residue.

[0380]

[0203] In certain embodiments, including any of the foregoing, L 2 is -(CR a Rb ) a -POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -CH2-POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -(CH2)2-POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -(CH2)3-POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -(CH2)4-POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -(CH2)5-POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -(CH2)6-POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -CR a R b -POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -(CR a R b )2-POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -(CR a R b )3-POLY 1 - is. In certain embodiments including any of the foregoing, L 2 is -(CR a R b )4-POLY 1 - is.

[0381]

[0204] In certain embodiments including any of the foregoing, L 2 is -POLY 1 - is.

[0382] In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -(CR a R b ) a -. In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -(CR a R b ) a -, and a is independently selected from 0, 1, 2, 3, 4, 5, or 6. In certain embodiments, including any of the foregoing, L 2 is -(CH2) a -POLY 1 -(CH2) a -, and a is independently selected from 0, 1, 2, 3, 4, 5, or 6. In certain embodiments, including any of the foregoing, L 2 is (CH2) a -POLY 1 -(CR a R b ) a -, and a is selected from 1, 2, 3, 4, 5, or 6. In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -(CH2) a (wherein a is independently selected from 0, 1, 2, 3, 4, 5, or 6).

[0383]

[0206] In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of a non-peptidic, hydrophilic polymer. In certain embodiments, POLY 1is a divalent residue of polyethylene glycol (PEG), poly(propylene glycol) (PPG), a copolymer of ethylene glycol and propylene glycol, poly(oxyethylated polyol), poly(olefin alcohol), poly(vinyl pyrrolidone), poly(hydroxyalkyl methacrylamide), poly(hydroxyalkyl methacrylate), polysaccharide, poly(α-hydroxy acid), poly(vinyl alcohol), polyphosphazene, polyoxazoline (POZ), poly(N-acryloylmorpholine), polysarcosine, or a combination thereof. In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of polyethylene glycol (PEG), poly(propylene glycol) (PPG), or a copolymer of ethylene glycol and propylene glycol.

[0384]

[0207] In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of polyethylene glycol (PEG). In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of poly(propylene glycol) (PPG). In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of a copolymer of ethylene glycol and propylene glycol. In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of poly(oxyethylated polyol). In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of poly(olefin alcohol). In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of poly(vinyl pyrrolidone). In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of poly(hydroxyalkyl methacrylamide). In certain embodiments, including any of the foregoing, POLY 1 is a divalent residue of poly(hydroxyalkyl methacrylate). In certain embodiments, including any of the foregoing, POLY1 is a divalent residue of a polysaccharide. In certain embodiments including any of the foregoing, POLY 1 is a divalent residue of a poly(α-hydroxy acid). In certain embodiments including any of the foregoing, POLY 1 is a divalent residue of poly(vinyl alcohol). In certain embodiments including any of the foregoing, POLY 1 is a divalent residue of a polyphosphazene. In certain embodiments including any of the foregoing, POLY 1 is a divalent residue of a polyoxazoline (POZ). In certain embodiments including any of the foregoing, POLY 1 is a divalent residue of poly(N-acryloylmorpholine). In certain embodiments including any of the foregoing, POLY 1 is a divalent residue of polysarcosine.

[0385]

[0208] In certain embodiments including any of the foregoing, POLY 1 is

[0386]

Chem.

[0387]

Chem.

[0388]

[0209] In some embodiments including any of the foregoing, R 5 is hydrogen. In some embodiments including any of the foregoing, R 5 is methyl.

[0389]

[0210] In certain embodiments including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -, where POLY 1 is

[0390]

Chem.

[0391]

Chem.

[0392]

Chem.

[0393]

Chem.

[0394]

Chemical formula

[0395]

Chemical formula

[0396]

Chemical formula

[0397]

Chemical formula

[0398]

Chem.

[0399]

[0211] In a specific embodiment including any of the foregoing, L 2 is

[0400]

Chem.

[0401]

Chem.

[0402]

Chem.

[0403]

Chem.

[0404]

Chem.

[0405]

Chemical formula

[0406]

Chemical formula

[0407]

Chemical formula

[0408]

[0212] In certain embodiments, including any of the foregoing, L 2 is

[0409] [Chemical formula] selected from. In certain embodiments including any of the foregoing, L 2 is

[0410] [Chemical formula] is. In certain embodiments including any of the foregoing, L 2 is

[0411] [Chemical formula] is.

[0412]

[0213] In certain embodiments including any of the foregoing, L 2 is

[0413] [Chemical formula] is. In certain embodiments including any of the foregoing, L 2 is

[0414] [Chemical formula] is. In certain embodiments including any of the foregoing, L 2 is

[0415] [Chemical formula] is. In certain embodiments including any of the foregoing, L 2 is

[0416] [Chemical formula] is. In certain embodiments, including any of the foregoing, L 2 is

[0417]

Chemical Formula

[0418]

[0214] In certain embodiments, including any of the foregoing, POLY 2 is a residue of a non-peptidic, hydrophilic polymer. In certain embodiments, POLY 2 is a residue of polyethylene glycol (PEG), methoxypolyethylene glycol (mPEG), poly(propylene glycol) (PPG), a copolymer of ethylene glycol and propylene glycol, poly(oxyethylated polyol), poly(olefin alcohol), poly(vinyl pyrrolidone), poly(hydroxyalkyl methacrylamide), poly(hydroxyalkyl methacrylate), polysaccharide, poly(α-hydroxy acid), poly(vinyl alcohol), polyphosphazene, polyoxazoline (POZ), poly(N-acryloylmorpholine), polysarcosine, or a combination thereof. In certain embodiments, including any of the foregoing, POLY 2 is a residue of polyethylene glycol (PEG), methoxypolyethylene glycol (mPEG), poly(propylene glycol) (PPG), or a copolymer of ethylene glycol and propylene glycol. In certain embodiments, including any of the foregoing, POLY 2 is a residue of methoxypolyethylene glycol (mPEG).

[0419]

[0215] In certain embodiments, including any of the foregoing, POLY 2 is a residue of polyethylene glycol (PEG). In certain embodiments, including any of the foregoing, POLY 2 is a residue of poly(propylene glycol) (PPG). In certain embodiments, including any of the foregoing, POLY 2is a residue of a copolymer of ethylene glycol and propylene glycol. In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(oxyethylated polyol). In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(olefin alcohol). In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(vinyl pyrrolidone). In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(hydroxyalkyl methacrylamide). In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(hydroxyalkyl methacrylate). In certain embodiments including any of the foregoing, POLY 2 is a residue of a polysaccharide. In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(α-hydroxy acid). In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(vinyl alcohol). In certain embodiments including any of the foregoing, POLY 2 is a residue of polyphosphazene. In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(oxazoline) (POZ). In certain embodiments including any of the foregoing, POLY 2 is a residue of poly(N-acryloylmorpholine). In certain embodiments including any of the foregoing, POLY 2 is a residue of polysarcosine.

[0420]

[0216] In certain embodiments including any of the foregoing, POLY 2 is

[0421]

Chemical formula

[0422] [Chemical formula] represents a bond to the remainder of the compound or conjugate. In certain embodiments, including any of the foregoing, x is an integer between 1 and 25. In certain embodiments, including any of the foregoing, x is an integer between 5 and 15. In some embodiments, including any of the foregoing, x is 1. In some embodiments, including any of the foregoing, x is 2. In some embodiments, including any of the foregoing, x is 3. In some embodiments, including any of the foregoing, x is 4. In some embodiments, including any of the foregoing, x is 5. In some embodiments, including any of the foregoing, x is 6. In some embodiments, including any of the foregoing, x is 7. In some embodiments, including any of the foregoing, x is 8. In some embodiments, including any of the foregoing, x is 9. In some embodiments, including any of the foregoing, x is 10. In some embodiments, including any of the foregoing, x is 11. In some embodiments, including any of the foregoing, x is 12. In some embodiments, including any of the foregoing, x is 13. In some embodiments, including any of the foregoing, x is 14. In some embodiments, including any of the foregoing, x is 15. In some embodiments, including any of the foregoing, x is 16. In some embodiments, including any of the foregoing, x is 17. In some embodiments, including any of the foregoing, x is 18. In some embodiments, including any of the foregoing, x is 19. In some embodiments, including any of the foregoing, x is 20. In certain embodiments, including any of the foregoing, x is an integer between 25 and 50. In certain embodiments, including any of the foregoing, x is an integer between 35 and 45. In certain embodiments, including any of the foregoing, x is an integer between 50 and 75. In certain embodiments, including any of the foregoing, x is an integer between 55 and 65. In certain embodiments, including any of the foregoing, x is an integer between 75 and 100. In certain embodiments, including any of the foregoing, x is an integer between 85 and 95. In certain embodiments, including any of the foregoing, x is an integer in the range of 1 to 25, 20 to 45, 40 to 65, 60 to 85, 70 to 95, or 75 to 100.

[0423]

[0217] In some embodiments including any of the foregoing, R 5 is hydrogen. In some embodiments including any of the foregoing, R 5 is methyl.

[0424]

[0218] In certain embodiments including any of the foregoing, L 2 is

[0425]

Chemical formula

[0426]

[0219] In certain embodiments including any of the foregoing, L 2 is

[0427]

Chemical formula

[0428]

[0220] In certain embodiments including any of the foregoing, L 2 is

[0429]

Chemical formula

[0430]

Chemical formula

[0431]

[0221] In certain embodiments including any of the foregoing, L 2 is

[0432] [Chemical formula] is. In certain embodiments, including any of the foregoing, L 2 is

[0433] [Chemical formula] is.

[0434]

[0222] In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-.

[0435]

[0223] In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-Z-(CR a R b ) a -Z-(CR a R b ) a -C(O)-. In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-NR 2 -(CR a R b ) a -NR 2 -(CR a R b ) a -C(O)-. In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-NH-(CR a R b ) a -NH-(CR a R b ) a -C(O)-. In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-NH-(CH2) a -NH-(CH2) a -C(O)-. In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-NH-(CH2)a -NH-(CH2) a is -C(O)-, and a is selected from 1, 2, and 3. In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-NH-CH2-NH-CH2-C(O)-

[0436]

[0224] In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-Z-(CR a R b ) a In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-NR 2 -(CH2) a In certain embodiments, including any of the foregoing, L 3 is -C(O)-AA-NH-(CH2)2

[0437]

[0225] In certain embodiments, including any of the foregoing, L 3 is -AA-. In certain embodiments, including any of the foregoing, L 3 is

[0438]

Chemical formula

[0439]

[0226] In certain embodiments, including any of the foregoing, -AA- is an amino acid residue. In certain embodiments, including any of the foregoing, -AA- is a peptide residue. In certain embodiments, including any of the foregoing, -AA- is a dipeptide residue, a tripeptide residue, a tetrapeptide residue, or a pentapeptide residue. In certain embodiments, including any of the foregoing, -AA- comprises at least one amino acid residue selected from alanine, glycine, valine, and asparagine. In certain embodiments, including any of the foregoing, -AA- comprises at least one amino acid residue selected from alanine and glycine. In certain embodiments, including any of the foregoing, -AA- is selected from the group consisting of

[0440]

Chemical formula

[0441]

[0227] In certain embodiments, including any of the foregoing, -AA- is

[0442]

Chemical formula

[0443]

[0228] In certain embodiments, including any of the foregoing, L 3 is -C(O)-.

[0444]

[0229] In certain embodiments, including any of the foregoing, L 3 is -C(O)-Z-(CR a R b ) a -C(O)-Z-L 4 -OC(O)-, and L 4 is

[0445]

Chemical formula

[0446]

[0230] In certain embodiments including any of the foregoing, L 4 is

[0447]

Chem.

[0448]

Chem.

[0449]

Chem.

[0450]

[0231] In some embodiments, including any of the foregoing, Su is a sugar moiety. In some embodiments, Su is a hexose form of a monosaccharide. Su can be a glucuronic acid or a mannose residue. In certain embodiments, including any of the foregoing, Su is

[0451]

Chemical formula

[0452]

Chemical formula

[0453]

Chemical formula

[0454]

Chemical formula

[0455]

[0232] In certain embodiments, including any of the foregoing, L 4 is

[0456]

Chemical formula

[0457]

[0233] In certain embodiments, including any of the foregoing, L 3 is -C(O)-NH-(CH2)2-C(O)-NH-L 4-OC(O)-, and L 4 is

[0458]

Chem.

[0459]

Chem.

[0460]

[0234] In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -, and L 3 is -C(O)-AA-. In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -; L 3 is -C(O)-AA-; POLY 1 is

[0461]

Chem.

[0462]

Chem.

[0463]

Chemical formula

[0464]

Chemical formula

[0465]

[0235] In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -; and L 3 is -C(O)-AA-Z-(CR a R b ) a -Z-(CR a R b ) a -C(O)-. In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -; and L 3 is -C(O)-AA-Z-(CR a R b ) a-Z-(CR a R b ) a -C(O)-; POLY 1 is

[0466]

Chem.

[0467]

Chem.

[0468]

Chem.

[0469]

Chem.

[0470]

[0236] In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -, and L 3 is -C(O). In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -; L 3 is -C(O); POLY 1 is

[0471]

Chemical formula

[0472]

Chemical formula

[0473]

Chemical formula

[0474]

Chemical formula

[0475]

[0237] In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 - and L 3 is absent. In certain embodiments, including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 - and L 3 is absent; POLY 1 is

[0476]

Chemical formula

[0477]

Chemical formula

[0478]

Chemical formula

[0479]

Chemical formula

[0480]

[0238] In certain embodiments including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -; and L 3 is -C(O)-AA-Z-(CR a R b ) a -. In certain embodiments including any of the foregoing, L 2 is -(CR a R b ) a -POLY 1 -; and L 3 is -C(O)-AA-Z-(CR a R b ) a -; and POLY 1 is

[0481]

Chemical formula

[0482]

Chemical formula

[0483]

Chemical formula

[0484]

Chemical formula

[0485]

[0239] -L 2 -L 3 - Non-limiting examples of include

[0486]

Chemical formula

[0487]

[0240] -L 2 -L 3 Additional non-limiting examples of - include

[0488]

Chemical formula

[0489]

[0241] In certain embodiments, including any of the foregoing, L 2 is

[0490]

Chemical formula

[0491]

Chemical formula

[0492]

Chemical formula

[0493] In certain embodiments, including any of the foregoing, L 2 is

[0494]

Chem.

[0495]

Chem.

[0496]

[0243] In certain embodiments, including any of the foregoing, L 2 is selected from the group consisting of

[0497]

Chem.

[0498]

[0244] In certain embodiments, including any of the foregoing, L 2 is selected from the group consisting of

[0499]

Chem.

[0500] In certain embodiments, including any of the foregoing, L 2 is

[0501]

Chem.

[0502]

Chem.

[0503] In certain embodiments, including any of the foregoing, L 2 is

[0504]

Chem.

[0505]

[0247] Non-limiting examples of -L 2 -L 3 - include

[0506]

Chem.

[0507]

[0248] In any embodiment of -L 2 -L 3 -, D can be

[0508]

Chem.

[0509]

[0249] -L 2 -L 3 Non-limiting examples of -D include

[0510]

Chem.

[0511]

Chem.

[0512]

Chem.

[0513]

Chem.

[0514]

Chem.

[0515]

[0250] In certain embodiments of Formulas (I)-(IH),

[0516]

Chem.

[0517]

Chem.

[0518]

Chem.

[0519] [Chemical formula] In certain embodiments of formulas (I)-(III),

[0520] [Chemical formula] is

[0521] [Chemical formula] In certain embodiments of formulas (I)-(III),

[0522] [Chemical formula] is

[0523] [Chemical formula] In certain embodiments of formulas (I)-(III),

[0524]

[0251] In certain embodiments of formulas (I)-(III),

[0525] [Chemical formula] is

[0526] [Chemical formula] and R a and R b are hydrogen. In certain embodiments of formulas (I)-(III),

[0527]

Chem.

[0528]

Chem.

[0529]

Chem.

[0530]

Chem.

[0531]

Chem.

[0532]

[0252] In certain embodiments of formulas (I) - (III),

[0533]

Chem.

[0534]

Chem.

[0535]

Chem.

[0536]

Chem.

[0537]

Chem.

[0538]

Chem.

[0539]

Chem.

[0540]

[0253] In certain embodiments of formulas (I) to (IH),

[0541]

Chem.

[0542]

Chem.

[0543]

Chemical formula

[0544]

Chemical formula

[0545]

Chemical formula

[0546]

Chemical formula

[0547]

Chemical formula

[0548]

[0254] In certain embodiments of formulas (I)-(IH),

[0549]

Chemical formula

[0550]

Chemical formula

[0551]

Chemical formula

[0552]

Chemical formula

[0553]

Chemical formula

[0554]

Chemical formula

[0555]

Chemical formula

[0556]

[0255]

[0557]

Chemical formula

[0558]

Chemical formula

[0559]

[0256] In certain embodiments of formula (III), (IIIA), or (IIIB),

[0560]

Chemical formula

[0561]

Chemical formula

[0562]

Chemical formula

[0563]

Chemical formula

[0564]

Chemical formula

[0565]

Chemical formula

[0566]

Chemical formula

[0567]

Chem.

[0568]

Chem.

[0569]

Chem.

[0570]

[0257] In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG is a group containing alkyne, cyclooctyne, strained alkene, tetrazine, amine, methylcyclopropene, thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains alkyne. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains cyclooctyne. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains strained alkene. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains tetrazine. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains amine. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains methylcyclopropene. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains thiol. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains para-acetyl-phenylalanine residue. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains oxyamine. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains maleimide. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG contains azide. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG is selected from the group consisting of

[0571]

Chemical formula

[0572]

Chemical formula

[0573]

Chem.

[0574]

Chem.

[0575]

Chem.

[0576]

Chem.

[0577]

Chem.

[0578]

Chem.

[0579]

Chem.

[0580] [Chem.] represents a bond to the remainder of the compound. In certain embodiments of Formulas (I)-(IH) or (III)-(IIIB), RG is

[0581] [Chem.] and

[0582] [Chem.] represents a bond to the remainder of the compound. In certain embodiments of Formulas (I)-(IH) or (III)-(IIIB), RG is

[0583] [Chem.] and R T is C 1~6 alkyl,

[0584] [Chem.] represents a bond to the remainder of the compound. In certain embodiments, R T is methyl, ethyl, or propyl. In certain embodiments, R T is methyl. In certain embodiments, R T is ethyl. In certain embodiments, R T is propyl. In certain embodiments, R T is butyl. In certain embodiments, R T is pentyl. In certain embodiments, R Tis hexyl. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG is

[0585]

Chemical formula

[0586]

Chemical formula

[0587]

Chemical formula

[0588]

Chemical formula

[0589]

Chemical formula

[0590]

Chemical formula

[0591]

Chemical formula

[0592] [Chemical formula] and

[0593] [Chemical formula] represents a bond to the remainder of the compound. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG is

[0594] [Chemical formula] and

[0595] [Chemical formula] represents a bond to the remainder of the compound. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG is -N3. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG is -NH2. In certain embodiments of formula (I)-(IIIB), RG is methylcyclopropene. In certain embodiments of formula (I)-(IH) or (III)-(IIIB), RG is -SH.

[0596]

[0258] In certain embodiments of formula (II)-(IIH),

[0597] [Chemical formula] is

[0598] [Chemical formula] and. In certain embodiments of formula (II)-(IIH),

[0599]

Chem.

[0600]

Chem.

[0601]

Chem.

[0602]

Chem.

[0603]

Chem.

[0604]

Chem.

[0605]

[0259] In certain embodiments of Formulas (II) - (IIH),

[0606]

Chem.

[0607]

Chem.

[0608]

Chem.

[0609]

Chem.

[0610]

Chem.

[0611]

Chem.

[0612]

Chem.

[0613]

[0260] In certain embodiments of Formulas (II) to (IIH),

[0614]

Chem.

[0615]

Chem.

[0616]

Chemical formula

[0617]

Chemical formula

[0618]

Chemical formula

[0619]

Chemical formula

[0620]

Chemical formula

[0621]

[0261] In certain embodiments of Formulas (II) - (IIH),

[0622]

Chemical formula

[0623]

Chem.

[0624]

Chem.

[0625]

Chem.

[0626]

Chem.

[0627]

Chem.

[0628]

Chem.

[0629]

[0262] In certain embodiments of formulas (II) to (IIH),

[0630]

Chem.

[0631]

Chem.

[0632]

[0263] In certain embodiments of formulas (II) to (IIH),

[0633]

Chem.

[0634]

Chem.

[0635]

Chem.

[0636]

Chem.

[0637]

Chem.

[0638]

[0264] In certain embodiments of formulas (IV) to (IVB),

[0639]

Chem.

[0640]

Chem.

[0641]

Chem.

[0642]

Chem.

[0643]

Chem.

[0644]

Chem.

[0645]

Chem.

[0646]

Chem.

[0647]

Chem.

[0648]

Chem.

[0649]

[0265] In certain embodiments of Formula (II)-(IIH), (IVA), or (IVB), RL is a group containing triazole, pyridazine, thiol, or oxime. In certain embodiments of Formula (II)-(IIH), (IV), (IVA), or (IVB), RL is a group containing triazole. In certain embodiments of Formula (II)-(IIH), (IV), (IVA), or (IVB), RL is a group containing pyridazine. In certain embodiments of Formula (II)-(IIH), (IV), (IVA), or (IVB), RL is a group containing thiol. In certain embodiments of Formula (II)-(IIH), (IV), (IVA), or (IVB), RL is a group containing oxime.

[0650]

[0266] In certain embodiments of Formula (II)-(IIH), (IV), (IVA), or (IVB), RL is

[0651]

Chem.

[0652]

Chem.

[0653]

Chem.

[0654]

Chem.

[0655]

Chem.

[0656]

Chem.

[0657]

Chem.

[0658]

Chem.

[0659]

Chem.

[0660]

Chem.

[0661]

Chem.

[0662]

Chem.

[0663]

[0267] In certain embodiments, including any of the foregoing, D is a cytotoxic payload selected from tubulin inhibitors, DNA topoisomerase I inhibitors, and DNA topoisomerase II inhibitors, or derivatives thereof. In some embodiments, including any of the foregoing, D is a tubulin inhibitor or a derivative thereof. In some embodiments, including any of the foregoing, D is a DNA topoisomerase I inhibitor or a derivative thereof. In some embodiments, including any of the foregoing, D is a DNA topoisomerase I inhibitor selected from the group consisting of irinotecan, SN-38, topotecan, and exatecan; or a derivative thereof. In some embodiments, including any of the foregoing, D is irinotecan or a derivative thereof. In some embodiments, including any of the foregoing, D is SN-38 or a derivative thereof. In some embodiments, including any of the foregoing, D is topotecan or a derivative thereof. In some embodiments, including any of the foregoing, D is exatecan or a derivative thereof. In some embodiments, including any of the foregoing, D is a DNA topoisomerase II inhibitor or a derivative thereof. In some embodiments, including any of the foregoing, D is a DNA topoisomerase II inhibitor selected from the group consisting of etoposide, teniposide, and telfluposide; or a derivative thereof. In some embodiments, including any of the foregoing, D is etoposide or a derivative thereof. In some embodiments, including any of the foregoing, D is teniposide or a derivative thereof. In some embodiments, including any of the foregoing, D is telfluposide or a derivative thereof. In some embodiments, including any of the foregoing, D is a payload selected from the group consisting of hemiasterlin, camptothecin, and anthracyclines; or derivatives thereof. Anthracyclines can include PNU-159682 and EDA PNU-159682 derivatives. In some embodiments, including any of the foregoing, D is an anthracycline selected from the group consisting of daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, and valrubicin; or a derivative thereof. In some embodiments, including any of the foregoing, D is daunorubicin or a derivative thereof.In some embodiments, including any of the foregoing, D is doxorubicin or a derivative thereof. In some embodiments, including any of the foregoing, D is epirubicin or a derivative thereof. In some embodiments, including any of the foregoing, D is idarubicin or a derivative thereof. In some embodiments, including any of the foregoing, D is mitoxantrone or a derivative thereof. In some embodiments, including any of the foregoing, D is valrubicin or a derivative thereof. In some embodiments, including any of the foregoing, D is hemiasterlin or a derivative thereof. In some embodiments, including any of the foregoing, D is camptothecin or a derivative thereof. In some embodiments, including any of the foregoing, D is an anthracycline or a derivative thereof. In some embodiments, including any of the foregoing, D is PNU-159682 or a derivative thereof. In some embodiments, including any of the foregoing, D is an EDA PNU compound or a derivative thereof. In some embodiments, including any of the foregoing, D is an EDA PNU-159682 derivative. In some embodiments, including any of the foregoing, D is hemiasterlin, exatecan, PNU-159682, or an EDA PNU-159682 derivative. In some embodiments, including any of the foregoing, D is hemiasterlin or a derivative thereof. In some embodiments, including any of the foregoing, D is exatecan or a derivative thereof. In some embodiments, including any of the foregoing, D is PNU-159682 or a derivative thereof. In some embodiments, including any of the foregoing, D is an EDA PNU-159682 compound or derivative. In some embodiments, including any of the foregoing, D is not an immunostimulatory compound.

[0664] In some embodiments, including any of the foregoing, D is an alkylating agent or a derivative thereof. In some embodiments, including any of the foregoing, D is a bifunctional alkylating agent or a derivative thereof. In some embodiments, including any of the foregoing, D is a bifunctional alkylating agent selected from the group consisting of cyclophosphamide, mechlorethamine, chlorambucil, and melphalan; or a derivative thereof. In some embodiments, including any of the foregoing, D is cyclophosphamide or a derivative thereof. In some embodiments, including any of the foregoing, D is mechlorethamine or a derivative thereof. In some embodiments, including any of the foregoing, D is chlorambucil or a derivative thereof. In some embodiments, including any of the foregoing, D is melphalan or a derivative thereof. In some embodiments, including any of the foregoing, D is a monofunctional alkylating agent or a derivative thereof. In some embodiments, including any of the foregoing, D is a monofunctional alkylating agent selected from the group consisting of dacarbazine, nitrosourea, and temozolomide; or a derivative thereof. In some embodiments, including any of the foregoing, D is dacarbazine or a derivative thereof. In some embodiments, including any of the foregoing, D is nitrosourea or a derivative thereof. In some embodiments, including any of the foregoing, D is temozolomide or a derivative thereof. In some embodiments, including any of the foregoing, D is a cytoskeletal disrupting agent (e.g., taxane) or a derivative thereof. In some embodiments, including any of the foregoing, D is a cytoskeletal disrupting agent selected from the group consisting of paclitaxel, docetaxel, abraxane, and taxotere; or a derivative thereof. In some embodiments, including any of the foregoing, D is paclitaxel or a derivative thereof. In some embodiments, including any of the foregoing, D is docetaxel or a derivative thereof. In some embodiments, including any of the foregoing, D is abraxane or a derivative thereof. In some embodiments, including any of the foregoing, D is taxotere or a derivative thereof. In some embodiments, including any of the foregoing, D is epothilone or a derivative thereof.In some embodiments, including any of the foregoing, D is an epothilone selected from the group consisting of epothilone A, epothilone B, epothilone C, epothilone D, and ixabepilone; or a derivative thereof. In some embodiments, including any of the foregoing, D is epothilone A or a derivative thereof. In some embodiments, including any of the foregoing, D is epothilone B or a derivative thereof. In some embodiments, including any of the foregoing, D is epothilone C or a derivative thereof. In some embodiments, including any of the foregoing, D is epothilone D or a derivative thereof. In some embodiments, including any of the foregoing, D is ixabepilone or a derivative thereof. In some embodiments, including any of the foregoing, D is a histone deacetylase inhibitor or a derivative thereof. In some embodiments, including any of the foregoing, D is a histone deacetylase inhibitor selected from the group consisting of vorinostat and romidepsin; or a derivative thereof. In some embodiments, including any of the foregoing, D is vorinostat or a derivative thereof. In some embodiments, including any of the foregoing, D is romidepsin or a derivative thereof. In some embodiments, including any of the foregoing, D is a kinase inhibitor or a derivative thereof. In some embodiments, including any of the foregoing, D is a kinase inhibitor selected from the group consisting of bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, and visomibep; or a derivative thereof. In some embodiments, including any of the foregoing, D is bortezomib or a derivative thereof. In some embodiments, including any of the foregoing, D is erlotinib or a derivative thereof. In some embodiments, including any of the foregoing, D is gefitinib or a derivative thereof. In some embodiments, including any of the foregoing, D is imatinib or a derivative thereof. In some embodiments, including any of the foregoing, D is vemurafenib or a derivative thereof. In some embodiments, including any of the foregoing, D is visomibep or a derivative thereof. In some embodiments, including any of the foregoing, D is a nucleotide analog and / or a precursor analog or a derivative thereof.In some embodiments, including any of the foregoing, D is a nucleotide analog and / or a precursor analog selected from the group consisting of azacitidine, azathioprine, capecitabine, cytarabine, doxifluridine, fluorouracil, gemcitabine, hydroxyurea, mercaptopurine, methotrexate, and tioguanine (formerly thioguanine); or a derivative thereof. In some embodiments, including any of the foregoing, D is azacitidine or a derivative thereof. In some embodiments, including any of the foregoing, D is azathioprine or a derivative thereof. In some embodiments, including any of the foregoing, D is capecitabine or a derivative thereof. In some embodiments, including any of the foregoing, D is cytarabine or a derivative thereof. In some embodiments, including any of the foregoing, D is doxifluridine or a derivative thereof. In some embodiments, including any of the foregoing, D is fluorouracil or a derivative thereof. In some embodiments, including any of the foregoing, D is gemcitabine or a derivative thereof. In some embodiments, including any of the foregoing, D is hydroxyurea or a derivative thereof. In some embodiments, including any of the foregoing, D is mercaptopurine or a derivative thereof. In some embodiments, including any of the foregoing, D is methotrexate or a derivative thereof. In some embodiments, including any of the foregoing, D is tioguanine (formerly thioguanine) or a derivative thereof. In some embodiments, including any of the foregoing, D is a peptide antibiotic or a derivative thereof. In some embodiments, including any of the foregoing, D is a peptide antibiotic selected from the group consisting of bleomycin and actinomycin; or a derivative thereof. In some embodiments, including any of the foregoing, D is bleomycin or a derivative thereof. In some embodiments, including any of the foregoing, D is actinomycin or a derivative thereof. In some embodiments, including any of the foregoing, D is a platinum-based payload or a derivative thereof.In some embodiments, including any of the foregoing, D is a platinum-based payload selected from the group consisting of carboplatin, cisplatin, and oxaliplatin; or a derivative thereof. In some embodiments, including any of the foregoing, D is carboplatin or a derivative thereof. In some embodiments, including any of the foregoing, D is cisplatin or a derivative thereof. In some embodiments, including any of the foregoing, D is oxaliplatin or a derivative thereof. In some embodiments, including any of the foregoing, D is a retinoid or a derivative thereof. In some embodiments, including any of the foregoing, D is a retinoid selected from the group consisting of tretinoin, alitretinoin, and bexarotene; or a derivative thereof. In some embodiments, including any of the foregoing, D is tretinoin or a derivative thereof. In some embodiments, including any of the foregoing, D is alitretinoin or a derivative thereof. In some embodiments, including any of the foregoing, D is bexarotene or a derivative thereof. In some embodiments, including any of the foregoing, D is a vinca alkaloid or a derivative thereof. In some embodiments, including any of the foregoing, D is a vinca alkaloid selected from the group consisting of vinblastine, vincristine, vindesine, vinorelbine; or a derivative thereof. In some embodiments, including any of the foregoing, D is vinblastine or a derivative thereof. In some embodiments, including any of the foregoing, D is vincristine or a derivative thereof. In some embodiments, including any of the foregoing, D is vindesine or a derivative thereof.

[0665]

[0269] In some embodiments, including any of the foregoing, D is

[0666]

Chemical formula

[0667]

[0270] In some embodiments, including any of the foregoing, D is

[0668] [Chemical formula] or selected from derivatives thereof.

[0669]

[0271] Non-limiting examples of the compounds of the present disclosure include

[0670] [Chemical formula]

[0671] [Chemical formula]

[0672] [Chemical formula]

[0673] [Chemical formula]

[0674] [Chemical formula]

[0675] [Chemical formula]

[0676] [Chemical formula]

[0677] [Chemical formula] include

[0678]

[0272] Representative compounds of the present disclosure are shown in Table A.

[0679]

Table 1-1

[0680]

Table 1-2

[0681]

Table 1-3

[0682]

Table 1-4

[0683]

Table 1-5

[0684]

Table 1-6

[0685]

Table 1-7

[0686]

[0273] Non-limiting examples of the conjugates of the present disclosure include

[0687]

Chemical formula

[0688]

Chem.

[0689]

Chem.

[0690]

Chem.

[0691]

Chem.

[0692]

Chem.

[0693]

Chem.

[0694]

Chem.

[0695]

Chem.

[0696]

Chem.

[0697]

Chem.

[0698]

Chem.

[0699]

Chem.

[0700]

Chem.

[0701]

[0274] Representative conjugates of the present disclosure are shown in Table B.

[0702]

Table 2-1

[0703]

Table 2-2

[0704]

Table 2-3

[0705]

Table 2-4

[0706]

Table 2-5

[0707]

Table 2-6

[0708]

Table 2-7

[0709]

Table 2-8

[0710]

Table 2-9

[0711]

Table 2-10

[0712]

Table 2-11

[0713]

Table 2-12

[0714]

Table 2-13

[0715]

[0275] Optically active compound In certain embodiments, the compounds provided herein may have several chiral centers, exist in optically active forms and racemic forms, and may be isolated. In certain embodiments, some compounds may exhibit polymorphism. One of ordinary skill in the art will recognize that the compounds provided herein can exist in any racemic, optically active, diastereomeric, polymorphic, positional isomeric, and / or stereoisomeric forms, and / or mixtures thereof.

[0716]

[0276] One of ordinary skill in the art will also recognize that such compounds described herein having the useful properties described herein are within the scope of this disclosure. One of ordinary skill in the art will further recognize methods for preparing the optically active forms of the compounds described herein, for example, by resolution of the racemate through recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or chromatographic separation using a chiral stationary phase. Further, most amino acids are chiral (i.e., the L-enantiomer is the configuration that occurs naturally as designated as L- or D-) and can exist as separate enantiomers.

[0717]

[0277] Examples of methods for obtaining optically active substances are known in the art and include at least the following: i) Physical separation of crystals - a technique for manually separating the macroscopic crystals of the individual enantiomers. This technique can be used when crystals of the separate enantiomers are present (i.e., the substance is an aggregate and the crystals are visually distinguishable); ii) Simultaneous crystallization - a technique in which the individual enantiomers crystallize separately from a solution of the racemate only when the solution of the racemate is an aggregate in the solid state; iii) Enzymatic resolution - a technique in which partial or complete separation of the racemate is achieved by the different reaction rates of the enantiomers in the presence of an enzyme; iv) Enzymatic asymmetric synthesis - a synthetic technique in which at least one step of the synthesis uses an enzymatic reaction to obtain an enantiomerically pure or enriched synthetic precursor of the desired enantiomer; v) Chemical asymmetric synthesis - A synthetic technique in which a desired enantiomer is synthesized from an achiral precursor using a chiral catalyst or a chiral auxiliary to generate chirality (i.e., chirality) in the product; vi) Diastereomer separation - A technique of treating a racemic compound with an enantiomerically pure reagent (chiral auxiliary) that converts individual enantiomers into diastereomers. The resulting diastereomers are then separated by chromatography or crystallization based on the more distinct differences in diastereomers, and then the chiral auxiliary is removed to obtain each enantiomer; vii) Primary and secondary asymmetric transformation - A technique in which the diastereomers of a racemate equilibrate in solution to result in the predominance of the diastereomers of the desired enantiomer, or the kinetic or thermodynamic crystallization of the diastereomers of the desired enantiomer disturbs the equilibrium so that, in principle, ultimately all substances are converted into the crystalline diastereomers of the desired enantiomer. The desired enantiomer is then derived from the diastereomer. viii) Kinetic optical resolution - This technique refers to the achievement of partial or complete resolution of a racemate (or further resolution of a partially resolved compound) by the unequal reaction rates of enantiomers with a chiral or non-racemic reagent or catalyst under kinetic conditions; ix) Enantioselective synthesis from non-racemic precursors - A synthetic technique in which the desired enantiomer is obtained from a chiral starting material and the stereochemical integrity is not impaired or is impaired only minimally during the synthesis process; x) Chiral liquid chromatography - A technique for separating the enantiomers of a racemate in a liquid mobile phase by different interactions with a stationary phase. To induce different interactions, the stationary phase can be made of a chiral material or the mobile phase can contain an additional chiral material; xi) Chiral gas chromatography - A technique for separating enantiomers by different interactions in a gas mobile phase using a column containing a fixed non-racemic adsorption phase by volatilizing the racemate; xii) Extraction with a chiral solvent - a technique for separating enantiomers by the kinetic or thermodynamic dissolution of a particular enantiomer in a specific chiral solvent; xiii) Transport through a chiral membrane - a technique in which a racemate is brought into contact with a thin film barrier. The barrier typically separates two miscible fluids, one of which contains the racemate, and a driving force such as a concentration or pressure difference causes preferential transport through the membrane barrier. Separation occurs as a result of the non-racemic nature of the membrane that allows only one enantiomer of the racemate to pass through.

[0718] In some embodiments, provided herein are compositions of the compounds of Formulas (I), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (III), (IIIA), and (IIIB), and conjugates of Formulas (II), (IIA), (IIB), (IIC), (IID), (IIE), (IIF), (IIG), (IIH), (IV), (IVA), and (IVB), that substantially exclude the designated stereoisomers of the compound or conjugate. In certain embodiments, in the methods, compounds, and conjugates of the disclosure, the compound or conjugate is substantially free of other stereoisomers. In some embodiments, the composition comprises a compound or conjugate that is at least 85%, 90%, 95%, 98%, or 99% to 100% by weight of the compound or conjugate, with the balance comprising other chemical species or enantiomers. In some embodiments, provided herein are compositions of the compounds of Formulas (I), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (III), (IIIA), and (IIIB), and conjugates of Formulas (II), (IIA), (IIB), (IIC), (IID), (IIE), (IIF), (IIG), (IIH), (IV), (IVA), and (IVB), that substantially exclude the designated enantiomer of the compound or conjugate. In certain embodiments, in the methods, compounds, and conjugates of the disclosure, the compound or conjugate is substantially free of other enantiomers. In some embodiments, the composition comprises a compound or conjugate that is at least 85%, 90%, 95%, 98%, or 99% to 100% by weight of the compound or conjugate, with the balance comprising other chemical species or enantiomers.

[0719]

[0279] Isotope-enriched compound Isotopically enriched compounds of formulas (I), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (III), (IIIA), and (IIIB) and conjugates of formulas (II), (IIA), (IIB), (IIC), (IID), (IIE), (IIF), (IIG), (IIH), (IV), (IVA), and (IVB), among others, are also provided herein.

[0720]

[0280] Isotopic enrichment (e.g., deuteration) of pharmaceuticals to improve pharmacokinetics (“PK”), pharmacodynamics (“PD”), and / or toxicity profiles has been previously demonstrated in some drug classes. See, e.g., Lijinsky et al., Food Cosmet. Toxicol., 20:393 (1982); Lijinsky et al., J. Nat. Cancer Inst., 69:1127 (1982); Mangold et al., Mutation Res. 308:33 (1994); Gordon et al., Drug Metab. Dispos., 15:589 (1987); Zello et al., Metabolism, 43:487 (1994); Gately et al., J. Nucl. Med., 27:388 (1986); Wade D, Chem. Biol. Interact. 117:191 (1999).

[0721]

[0281] Isotopic enrichment of a drug can be used to, for example, (1) reduce or eliminate an unwanted metabolite; (2) increase the half-life of the parent drug; (3) decrease the number of administrations necessary to achieve a desired effect; (4) decrease the amount of the administration necessary to achieve a desired effect; (5) increase the formation of an active metabolite, if formed; and / or (6) decrease the production of a harmful metabolite in a particular tissue. Isotopic enrichment of a drug can also be used to create a more effective and / or safer drug for combination therapy, whether the combination therapy is intended or not.

[0722]

[0282] Replacing an atom with one of its isotopes often changes the reaction rate of a chemical reaction. This phenomenon is known as the kinetic isotope effect (“KIE”). For example, if a C-H bond is broken during the rate-determining step of a chemical reaction (i.e., the step with the highest transition state energy), replacing the reactive hydrogen with a (heavier) isotope causes a decrease in the reaction rate. The deuterium kinetic isotope effect (“DKIE”) is the most common form of the KIE (see, e.g., Foster et al., Adv. Drug Res., Vol. 14, pp. 1-36 (1985); Kushner et al., Can. J. Physiol. Pharmacol., Vol. 77, pp. 79-88 (1999)).

[0723]

[0283] The magnitude of the DKIE can be expressed as the ratio between the rate of a given reaction in which a C-H bond is broken and the rate of the same reaction in which deuterium replaces hydrogen and a C-D bond is broken. The DKIE can range from about 1 (no isotope effect) to very large numbers, e.g., up to 50 or more, meaning that the reaction can be 50 times or more slower when deuterium replaces hydrogen.

[0724]

[0284] Replacing hydrogen with tritium (“T”) gives a stronger bond than deuterium and a numerically larger isotope effect. Similarly, but not limited thereto, substitution with isotopes of other elements including substitution of carbon with 13 C or 14 C; substitution of sulfur with 33 S, 34 S, or 36 S; substitution of nitrogen with 15 N; and substitution of oxygen with 17 O or 18 O can result in a similar kinetic isotope effect.

[0725]

[0285] The body of an animal expresses various enzymes from its circulatory system for the purpose of eliminating foreign substances such as therapeutic agents. Examples of such enzymes include cytochrome P450 enzymes ("CYPs"), esterases, proteases, reductases, dehydrogenases, and monoamine oxidases that react with these foreign substances and convert them into more polar intermediates or metabolites for renal excretion. Some of the most common metabolic reactions of pharmaceutical compounds involve oxidation of a carbon - hydrogen (C - H) bond to either a carbon - oxygen (C - O) or a carbon - carbon (C = C) π - bond. The resulting metabolites can be stable or unstable under physiological conditions and can have substantially different PK / PD, as well as acute and long - term toxicity profiles compared to the parent compound. For many drugs, such oxidation is rapid. Thus, these drugs often require multiple or high daily doses.

[0726]

[0286] Thus, the isotope enrichment at certain positions of the compounds provided herein results in a detectable KIE that affects the pharmacological, PK, PD, and / or toxicological profiles of the compounds provided herein compared to similar compounds having a natural isotope composition.

[0727]

[0287] The conjugates of formula (II), (IIA), (IIB), (IIC), (IID), (IIE), (IIF), (IIG), (IIH), (III), (IIIA), and (IIIB) Conjugates of polymers with one of the compounds of formulas (I)-(IH), (III), (IIIA), and (IIIB) described herein are provided herein. The conjugates are covalently linked either directly or indirectly through a linker. In certain embodiments, the conjugate comprises a polymer conjugated with one or more compounds of formulas (I)-(IH), (III), (IIIA), and (IIIB) described herein. In certain embodiments, the conjugate comprises two or more polymers. In certain embodiments, the polymer is linked to one, two, three, four, five, six, seven, eight, or more compounds of formulas (I)-(IH), (III), (IIIA), and (IIIB).

[0728]

[0288] The linker can be any linker capable of forming at least one bond with the polymer and at least one bond with the compounds of formulas (I)-(IH), (III), (IIIA), and (IIIB). Useful linkers are described herein, particularly in the following sections and examples.

[0729]

[0289] The polymer can be any polymer considered suitable by one of ordinary skill in the art. In certain embodiments, the polymer is a second compound. In certain embodiments, COMP is a residue of the second compound. In certain embodiments, the polymer is a protein, peptide, antibody or antigen-binding fragment thereof, nucleic acid, carbohydrate, or other large molecule composed of polymerized monomers. In certain embodiments, the polymer is a peptide of two or more residues. In certain embodiments, the polymer is a peptide of ten or more residues. In certain embodiments, the polymer has a mass of at least 1000 Da. In certain embodiments, the polymer contains at least 1000 atoms. Useful polymers are described in the following sections.

[0730]

[0290] Polymer (COMP) The polymer (COMP) can be any polymer that is considered suitable by those skilled in the art. In certain embodiments, the polymer is a protein, peptide, antibody or antigen-binding fragment thereof, nucleic acid, carbohydrate, or other large molecule composed of polymerized monomers. In certain embodiments, the polymer is a protein. In certain embodiments, the polymer is an antibody or antigen-binding fragment thereof. In some embodiments, COMP is a residue of a polypeptide. In some embodiments, COMP is a residue of an antibody. In some embodiments, COMP is a residue of an antibody chain.

[0731]

[0291] In some embodiments, the macromolecule is an antibody or an antigen-binding fragment thereof. In some embodiments, the macromolecule is a known antibody. Useful antibodies include, but are not limited to, Rituxan® (IDEC / Genentech / Roche) (see, e.g., U.S. Patent No. 5,736,137), a chimeric anti-CD20 antibody approved for treating non-Hodgkin's lymphoma; HuMax-CD20, an anti-CD20 antibody currently being developed by Genmab, the anti-CD20 antibody described in U.S. Patent No. 5,500,362, AME-133 (Applied Molecular Evolution), hA20 (Immunomedics, Inc.), HumaLYM (Intracel), and PRO70769 (PCT Application No. PCT / US2003 / 040426); trastuzumab (Herceptin® (Genentech)) (see, e.g., U.S. Patent No. 5,677,171), a humanized anti-HER2 / neu antibody approved for treating breast cancer; pertuzumab (rhuMab-2C4, Omnitarg®), currently being developed by Genentech; an anti-HER2 antibody (U.S. Patent No. 4,753,894); cetuximab (Erbitux® (Imclone)) (U.S. Patent No. 4,943,533; PCT Publication No. WO 96 / 40210), a chimeric anti-EGFR antibody in clinical trials for various cancers; ABX-EGF (U.S. Patent No. 6,235,883), currently being developed by Abgenix-Immunex-Amgen; HuMax-EGFr (U.S. Patent No. 7,247,301), currently being developed by Genmab; 425, EMD55900, EMD62000, and EMD72000 (Merck KGaA) (U.S. Patent No. 5,558,864; Murthy et al. (1987) Arch. Biochem. Biophys. 252(2):549-60; Rodeck et al. (1987) J. Cell Biochem. 35(4):315-20; Kettleborough et al. (1991) Protein Eng. 4(7):773-83); ICR62 (Institute of Cancer Research) (PCT Publication No.WO 95 / 20045; Modjtahedi et al. (1993) J. Cell. Biophys. 22(I-3):129-46; Modjtahedi et al. (1993) Br. J. Cancer 67(2):247-53; Modjtahedi et al. (1996) Br. J. Cancer 73(2): 228-35; Modjtahedi et al. (2003) Int. J. Cancer 105(2):273-80); TheraCIM hR3 (YM Biosciences, Canada and Centro de Immunologia Molecular, Cuba (US Patent No. 5,891,996; US Patent No. 6,506,883; Mateo et al. (1997) Immunotechnol. 3(1):71-81); mAb-806 (Ludwig Institute for Cancer Research, Memorial Sloan-Kettering) (Jungbluth et al. (2003) Proc. Natl. Acad. Sci. USA. 100(2):639-44); KSB-102 (KS Biomedix); MR1-1 (IVAX, National Cancer Institute) (PCT Publication No. WO 01 / 62931); and SC100 (Scancell) (PCT Publication No. WO 01 / 88138); alemtuzumab (Campath®, Millennium), a humanized mAb currently approved for the treatment of B-cell chronic lymphocytic leukemia; muromonab-CD3 (Orthoclone OKT3®), an anti-CD3 antibody developed by Ortho Biotech / Johnson & Johnson, ibritumomab tiuxetan (Zevalin®), an anti-CD20 antibody developed by IDEC / Schering AG, gemtuzumab ozogamicin (Mylotarg®), an anti-CD33 (p67 protein) antibody developed by Celltech / Wyeth, alefacept (Amevive®), an anti-LFA-3 developed by BiogenFc fusion proteins), abciximab (ReoPro®), basiliximab (Simulect®) developed by Centocor / Lilly, palivizumab (Synagis®) developed by Novartis, infliximab (Remicade®) developed by Medimmune, anti-TNFα antibodies developed by Centocor, adalimumab (Humira®), anti-TNFα antibodies developed by Abbott, Humicade®, anti-TNFα antibodies developed by Celltech, golimumab (CNTO-148), fully human TNF antibodies developed by Centocor, etanercept (Enbrel®), p75 TNF receptor Fc fusion proteins developed by Immunex / Amgen, Ienercept, p55 TNF receptor Fc fusion proteins previously developed by Roche, ABX-CBL, anti-CD147 antibodies developed by Abgenix, ABX-IL8, anti-IL8 antibodies developed by Abgenix, ABX-MA1, anti-MUC18 antibodies developed by Abgenix, pemtumomab (R1549, 90Y-muHMFG1), anti-MUC1 under development by Antisoma, Therex (R1550), anti-MUC1 antibodies developed by Antisoma, AngioMab (AS1405), developed by Antisoma, HuBC-1, developed by Antisoma, thioplatin (AS1407) developed by Antisoma, Antegren® (natalizumab), anti-α4β1 (VLA-4) and α4β7 antibodies developed by Biogen, VLA-1 mAb, anti-VLA-1 integrin antibodies developed by Biogen, LTBR mAb, anti-lymphotoxin β receptor (LTBR) antibodies developed by Biogen, CAT-152, anti-TGF-β antibodies developed by Cambridge Antibody Technology, ABT 874 (J695), anti-IL-12 p40 antibodies developed by Abbott, CAT-192, CambridgeThe anti-TGFβ1 antibody, CAT-213, developed by Antibody Technology and Genzyme; the anti-eotaxin 1 antibody, LymphoStat-B®, developed by Cambridge Antibody Technology; the anti-Blys antibody, TRAIL-R1 mAb, developed by Cambridge Antibody Technology and Human Genome Sciences Inc.; the anti-TRAIL-R1 antibody, Avastin® (bevacizumab, rhuMAb-VEGF), the anti-VEGF antibody developed by Genentech; the anti-HER receptor family antibody developed by Genentech; anti-tissue factor (ATF), the anti-tissue factor antibody developed by Genentech; Xolair® (omalizumab), the anti-IgE antibody developed by Genentech; Raptiva® (efalizumab), the anti-CD11a antibody developed by Genentech and Xoma; the MLN-02 antibody (formerly LDP-02), developed by Genentech and Millennium Pharmaceuticals; HuMax CD4, the anti-CD4 antibody developed by Genmab; HuMax-IL15, the anti-IL15 antibody developed by Genmab and Amgen; HuMax-Inflam, developed by Genmab and Medarex; HuMax-Cancer, the anti-heparanase I antibody developed by Genmab, Medarex and Oxford GlycoSciences; HuMax-Lymphoma, developed by Genmab and Amgen; HuMax-TAC, developed by Genmab; IDEC-131, and the anti-CD40L antibody developed by IDEC Pharmaceuticals; IDEC-151 (clenoliximab), the anti-CD4 antibody developed by IDEC Pharmaceuticals; IDEC-114, IDECThe anti-CD80 antibody developed by Pharmaceuticals, IDEC-152, the anti-CD23 developed by IDEC Pharmaceuticals, the anti-macrophage migration inhibitory factor (MIF) antibody developed by IDEC Pharmaceuticals, BEC2, the anti-idiotype antibody developed by Imclone, IMC-1C11, the anti-KDR antibody developed by Imclone, DC101, the anti-flk-1 antibody developed by Imclone, the anti-VE cadherin antibody developed by Imclone, CEA-Cide® (Racotumomab), the anti-carcinoembryonic antigen (CEA) antibody developed by Immunomedics, LymphoCide® (Epratuzumab), the anti-CD22 antibody developed by Immunomedics, AFP-Cide, developed by Immunomedics, MyelomaCide, developed by Immunomedics, LkoCide, developed by Immunomedics, ProstaCide, developed by Immunomedics, MDX-010, the anti-CTLA4 antibody developed by Medarex, MDX-060, the anti-CD30 antibody developed by Medarex, MDX-070 developed by Medarex, MDX-018 developed by Medarex, Osidem® (IDM-1), as well as Medarex and Immuno-DesignedThe anti-HER2 antibody developed by Molecules, HuMax®-CD4, the anti-CD4 antibody developed by Medarex and Genmab, HuMax-IL15, the anti-IL15 antibody developed by Medarex and Genmab, CNTO148, the anti-TNFα antibody developed by Medarex and Centocor / J&J, CNTO1275, the anti-cytokine antibody developed by Centocor / J&J, MOR101 and MOR102, the anti-intercellular adhesion molecule-1 (ICAM-1) (CD54) antibody developed by MorphoSys, MOR201, the anti-fibroblast growth factor receptor 3 (FGFR-3) antibody developed by MorphoSys, Nuvion® (visilizumab), the anti-CD3 antibody developed by Protein Design Labs, HuZAF®, the anti-gamma interferon antibody developed by Protein Design Labs, anti-α5β1 integrin, developed by Protein Design Labs, anti-IL-12, developed by Protein Design Labs, ING-1, the anti-Ep-CAM antibody developed by Xoma antibodies, Xolair® (omalizumab), the humanized anti-IgE antibody developed by Genentech and Novartis, and MLN01, the anti-β2 integrin antibody developed by Xoma are included.

[0732]

[0292] In another embodiment, the therapeutic agent is KRN330 (Kirin); huA33 antibody (A33, Ludwig Institute for Cancer Research); CNTO95 (αV integrin, Centocor); MEDI-522 (αVβ3 integrin, Medimmune); brolocizumab (αVβ1 integrin, Biogen / PDL); human mAb 216 (B cell glycosylation epitope, NCl); BiTE MT103 (bispecific CD19×CD3, Medimmune); 4G7×H22 (bispecific B cell×FcγR1, Medarex / Merck Kga); rM28 (bispecific CD28×MAPG, European Patent No. 1444268); MDX447 (EMD82633) (bispecific CD64×EGFR, Medarex); catumaxomab (removab) (bispecific EpCAM×anti-CD3, Trion / Fres); ertumaxomab (bispecific HER2 / CD3, Fresenius Biotech); oregovomab (OvaRex) (CA-125, ViRexx); Rencarex® (WXG250) (carbonic anhydrase IX, Wilex); CNTO888 (CCL2, Centocor); TRC105 (CD105 (endoglin), Tracon); BMS-663513 (CD137 agonist, Bristol Myers Squibb); MDX-1342 (CD19, Medarex); sipuleucel-T (MEDI-507) (CD2, Medimmune); ofatumumab (Humax-CD20) (CD20, Genmab); rituximab (Rituxan) (CD20, Genentech); belzutifan (hA20) (CD20, Immunomedics); epratuzumab (CD22, Amgen); lumiliximab (IDEC152) (CD23, Biogen); muromonab-CD3 (CD3, Ortho); HuM291 (CD3 fc receptor, PDL Biopharma); HeFi-1, CD30, NCl); MDX-060 (CD30, Medarex); MDX-1401 (CD30, Medarex); SGN-30 (CD30, Seattle Genentics);SGN-33 (Rituximab) (CD33, Seattle Genetics); Zanolimumab (HuMax-CD4) (CD4, Genmab); HCD122 (CD40, Novartis); SGN-40 (CD40, Seattle Genetics); MabCampath (Alemtuzumab) (CD52, Genzyme); MDX-1411 (CD70, Medarex); hLL1 (EPB-1) (CD74.38, Immunomedics); Galiximab (IDEC-144) (CD80, Biogen); MT293 (TRC093 / D93) (Cleaved Collagen, Tracon); HuLuc63 (CS1, PDL Pharma); Ipilimumab (MDX-010) (CTLA4, Bristol Myers Squibb); Tremelimumab (Ticilimumab, CP-675,2) (CTLA4, Pfizer); HGS-ETR1 (Mapatumumab) (DR4 TRAIL-R1 Agonist, Human Genome Science / Glaxo Smith Kline); AMG-655 (DR5, Amgen); Apomab (Apomab) (DR5, Genentech); CS-1008 (DR5, Daiichi Sankyo); HGS-ETR2 (Lexatumumab) (DR5 TRAIL-R2 Agonist, HGS); Cetuximab (Erbitux) (EGFR, Imclone); IMC-11F8 (EGFR, Imclone); Nimotuzumab (EGFR, YM Bio); Panitumumab (Vectabix) (EGFR, Amgen); Zalutumumab (HuMaxEGFr) (EGFR, Genmab); CDX-110 (EGFRvIII, AVANT Immunotherapeutics); Adecatumumab (MT201) (Epcam, Merck); Edrecolomab (Panorex, 17-1A) (Epcam, Glaxo / Centocor); MORAb-003 (Folate Receptor a, Morphotech); KW-2871 (Ganglioside GD3, Kyowa); MORAb-009 (GP-9, Morphotech); CDX-1307 (MDX-1307) (hCGb, Celldex); Trastuzumab (Herceptin) (HER2, Celldex);Pertuzumab (rhuMAb 2C4) (HER2 (DI), Genentech); Apolizumab (HLA-DRβ chain, PDL Pharma); AMG-479 (IGF-1R, Amgen); Anti-IGF-1R R1507 (IGF1-R, Roche); CP 751871 (IGF1-R, Pfizer); IMC-A12 (IGF1-R, Imclone); BIIB022 (IGF-1R, Biogen); Mik-β-1 (IL-2Rb (CD122), Hoffman-La Roche); CNTO328 (IL6, Centocor); Anti-KIR (1-7F9) (killer cell Ig-like receptor (KIR), Novo); Hu3S193 (Lewis (y), Wyeth, Ludwig Institute of Cancer Research); hCBE-11 (LTβR, Biogen); HuHMFG1 (MUC1, Antisoma / NCl); RAV12 (N-linked carbohydrate epitope, Raven); CAL (parathyroid hormone-related protein (PTH-rP), University of California); CT-011 (PD1, CureTech); MDX-1106 (ono-4538) (PD1, Medarex / Ono); Mab CT-011 (PD1, Curetech); IMC-3G3 (PDGFRa, Imclone); Bavituximab (phosphatidylserine, Peregrine); huJ591 (PSMA, Cornell Research Foundation); muJ591 (PSMA, Cornell Research Foundation); GC1008 (TGFb (pan) inhibitor (IgG4), Genzyme); Infliximab (Remicade) (TNFa, Centocor); A27.15 (transferrin receptor, Salk Institute, INSERN International Publication No. 2005 / 111082); E2.3 (transferrin receptor, Salk Institute); Bevacizumab (Avastin) (VEGF, Genentech); HuMV833 (VEGF, Tsukuba Research Lab, PCT Publication No. International Publication No. 2000 / 034337, University of Texas); IMC-18F1 (VEGFR1, Imclone);It includes IMC-1121 (VEGFR2, Imclone).;

[0733]

[0293] Examples of useful bispecific antibodies include, but are not limited to, bispecific antibodies having one antibody directed against a tumor cell antigen and the other antibody directed against a cytotoxic agent molecule, such as anti-FcγRI / anti-CD15, anti-p185 HER2 / FcγRIII (CD16), anti-CD3 / anti-malignant B cell (1D10), anti-CD3 / anti-p185 HER2 , anti-CD3 / anti-p97, anti-CD3 / anti-renal cell carcinoma, anti-CD3 / anti-OVCAR-3, anti-CD3 / L-D1 (anti-colorectal cancer), anti-CD3 / anti-melanocyte stimulating hormone analog, anti-EGF receptor / anti-CD3, anti-CD3 / anti-CAMA1, anti-CD3 / anti-CD19, anti-CD3 / MoV18, anti-neural cell adhesion molecule (NCAM) / anti-CD3, anti-folate binding protein (FBP) / anti-CD3, anti-pan-cancer associated antigen (AMOC-31) / anti-CD3; bispecific antibodies having one antibody that specifically binds to a tumor antigen and another antibody that binds to a toxin, such as anti-saponin / anti-Id-1, anti-CD22 / anti-saponin, anti-CD7 / anti-saponin, anti-CD38 / anti-saponin, anti-CEA / anti-ricin A chain, anti-interferon α (IFN-α) / anti-hybridoma idiotype, anti-CEA / anti-vinca alkaloid; bispecific antibodies that convert an enzyme-activated prodrug, such as anti-CD30 / anti-alkaline phosphatase (catalyzing the conversion of mitomycin phosphate prodrug to mitomycin alcohol); bispecific antibodies that can be used as fibrinolytic agents, such as anti-fibrin / anti-tissue plasminogen activator (tPA), anti-fibrin / anti-urokinase type plasminogen activator (uPA); bispecific antibodies for targeting immune complexes to cell surface receptors, such as anti-low density lipoprotein (LDL) / anti-Fc receptor (e.g., FcγRI, FcγRII, or FcγRIII); bispecific antibodies for use in the treatment of infectious diseases, such as anti-CD3 / anti-simple herpes virus (HSV), anti-T cell receptor:CD3 complex / anti-influenza, anti-FcγR / anti-HIV; bispecific antibodies for detecting tumors in vitro or in vivo, such as anti-CEA / anti-EOTUBE, anti-CEA / anti-DPTA, anti-anti-p185HER2 / anti-hapten; bispecific antibodies as vaccine adjuvants (see Fanger, M W et al., Crit Rev Immunol. 1992;12(34):101-24, incorporated herein by reference); and bispecific antibodies as diagnostic tools, such as anti-rabbit IgG / anti-ferritin, anti-horseradish peroxidase (HRP) / anti-hormone, anti-somatostatin / anti-substance P, anti-HRP / anti-FITC, anti-CEA / anti-β-galactosidase (see Nolan, O. and O’Kennedy, R., Biochim Biophys Acta. August 1, 1990;1040(1):1-11, incorporated herein by reference). Examples of trispecific antibodies include anti-CD3 / anti-CD4 / anti-CD37, anti-CD3 / anti-CD5 / anti-CD37, and anti-CD3 / anti-CD8 / anti-CD37.

[0734]

[0294] Conjugation In certain embodiments, conjugates can be formed from polymers that include one or more reactive groups. In certain embodiments, conjugates can be formed from polymers that include all naturally encoded amino acids. One of ordinary skill in the art will recognize that some naturally encoded amino acids contain reactive groups that can conjugate with the compounds or linkers of Formulas (I), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (III), (IIIA), and (IIIB). These reactive groups include cysteine side chains, lysine side chains, and amino terminal groups. In these embodiments, the conjugate can include a compound or linker of Formulas (I), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (III), (IIIA), and (IIIB) linked to a residue of an antibody reactive group. In these embodiments, the precursor of a compound of Formulas (I), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (III), (IIIA), and (IIIB) or a linker precursor includes a reactive group that can form a bond with a reactive group of an antibody or an antigen-binding fragment thereof. Exemplary reactive groups include maleimide groups, activated carbonates (including, but not limited to, p-nitrophenyl esters), and activated esters (including, but not limited to, N-hydroxysuccinimide, p-nitrophenyl esters, and aldehydes). Particularly useful reactive groups include maleimide and succinimide, such as N-hydroxysuccinimide, for forming bonds with cysteine and lysine side chains. Additional reactive groups are described in the following sections and examples.

[0735]

[0295] Reactive group The reactive group promotes conjugation with a second compound such as a polymer (i.e., COMP) described herein of a compound of formula (I)-(IH), (III), (IIIA), or (IIIB) described herein to form a conjugate of formula (II)-(IIH), (III), (IIIA), or (IIIB) described herein. In certain embodiments, the reactive group is designated as RG herein. The reactive group can react via any suitable reaction mechanism known to those skilled in the art. In certain embodiments, the reactive group (RG) reacts through a [3+2] alkyne-azide cycloaddition reaction, inverse electron demand Diels-Alder ligation reaction, thiol-electrophile reaction, or carbonyl-oxyamine reaction as described in detail herein. In certain embodiments, the reactive group (RG) comprises an alkyne, strained alkyne, tetrazine, thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide. In certain embodiments, the reactive group is,

[0736] [Chemical formula] , -N3, or SH; R T is lower alkyl. In certain embodiments, R T is methyl, ethyl, or propyl. In some embodiments, R T is methyl. In some embodiments, R T is ethyl. In some embodiments, R T is propyl. Additional reactive groups are described, for example, in U.S. Patent Application Publication No. 2014 / 0356385, U.S. Patent Application Publication No. 2013 / 0189287, U.S. Patent Application Publication No. 2013 / 0251783, U.S. Patent No. 8,703,936, U.S. Patent No. 9,145,361, U.S. Patent No. 9,222,940, and U.S. Patent No. 8,431,558.

[0737]

[0296] After conjugation, a divalent residue of the reactive group (referred to herein as RL) is formed and is attached to the residue of the second compound (e.g., COMP). The structure of the divalent residue is determined by the type of conjugation reaction employed to form the conjugate.

[0738]

[0297] [3+2] alkyne-azide cycloaddition reaction

[0739]

Chem.

[0740]

[0298] Advantageously, the compounds described herein that contain a conjugated alkyne group or an azide group facilitate a selective and efficient reaction with a second compound that contains a complementary azide group or alkyne group. The azide group and the alkyne group are thought to react in a 1,3-dipolar cycloaddition reaction to form a 1,2,3-triazolylene moiety that links the compound described herein that contains an alkyne group or an azide group to the second compound. This reaction between the azide and the alkyne to form a triazole is commonly known to those skilled in the art as the Huisgen cycloaddition reaction or the [3+2] alkyne-azide cycloaddition reaction.

[0741]

[0299] The unique reactivity of azide and alkyne functional groups makes them useful for the selective modification of polypeptides and other biological molecules. Organic azides, particularly aliphatic azides, and alkynes are generally stable to common reaction chemical conditions. In particular, both azide and alkyne functional groups are inert to the side chains of the 20 common amino acids found in naturally occurring polypeptides. When brought into proximity, the "spring-loaded" nature of the azide and alkyne groups becomes apparent, and it is thought that the azide and alkyne groups react selectively and efficiently via a [3+2] alkyne-azide cycloaddition reaction to produce the corresponding triazole. See, for example, Chin J. et al., Science 301:964-7 (2003); Wang, Q. et al., J. Am. Chem. Soc. 125, 3192-3193 (2003); Chin, J.W. et al., J. Am. Chem. Soc. 124:9026-9027 (2002).

[0742]

[0300] The [3+2] alkyne-azide cycloaddition reaction involves a selective cycloaddition reaction rather than a nucleophilic substitution [see, for example, Padwa, A., COMPREHENSIVE ORGANIC SYNTHESIS, Volume 4, (Editor Trost, B.M., 1991), pages 1069-1109; Huisgen, R., 1,3-DIPOLAR CYCLOADDITION CHEMISTRY, (Editor Padwa, A., 1984), pages 1-176]. By incorporating non-naturally encoded amino acids having azide-containing side chains and alkyne-containing side chains, the resulting polypeptide can be selectively modified at the positions of non-naturally encoded amino acids. The cycloaddition reaction involving an azide-containing compound or an alkyne-containing compound can be carried out at room temperature under aqueous conditions by adding Cu(II) (including, but not limited to, a catalytic amount of CuSO4) in the presence of a catalytic amount of a reducing agent for reducing Cu(II) to Cu(I) in situ. See, for example, Wang, Q. et al., J. Am. Chem. Soc. 125, 3192-3193 (2003); Tornoe, C. W. et al., J. Org. Chem. 67:3057-3064 (2002); Rostovtsev et al., Angew. Chem. Int. Ed. 41:2596-2599 (2002). Exemplary reducing agents include, but are not limited to, ascorbic acid, metallic copper, quinines, hydroquinone, vitamin K, glutathione, cysteine, Fe 2+ , Co 2+ , and application of a potential.

[0743]

[0301] In certain embodiments, when the conjugate is formed through a [3+2] alkyne-azide cycloaddition reaction, the divalent residue of the reactive group (e.g., RL) includes a triazole ring or a fused cyclic group containing a triazole ring. In certain embodiments, when the conjugate is formed through a strain-promoted [3+2] alkyne-azide cycloaddition (SPAAC) reaction, the divalent residue of the reactive group (e.g., RL) is

[0744]

Chemical Structure

[0745]

[0302] When the conjugate of formula (II) - (IIH), (IV), (IVA), or (IVB) is formed by [3 + 2] alkyne - azide cycloaddition, the conjugate includes both regioisomers. In certain embodiments, the conjugate of formula (II) - (IIH), (IV), (IVA), or (IVB) is a mixture of regioisomers formed from [3 + 2] alkyne - azide cycloaddition.

[0746]

[0303] Inverse electron demand ligation reaction

[0747]

Chemical formula

[0748]

[0304] Advantageously, compounds containing a terminal tetrazine or strained alkene group promote a selective and efficient reaction with a second compound containing a strained alkene or tetrazine group. Tetrazine and strained alkene are thought to react in an inverse demand Diels - Alder reaction followed by a retro - Diels - Alder reaction that links a compound containing a terminal tetrazine or strained alkene group to the second compound. The reaction is thought to be highly specific with little to no cross - reactivity with functional groups within biomolecules. The reaction can be carried out under mild conditions, e.g., at room temperature without a catalyst. This reaction between tetrazine and strained alkene is generally known to those skilled in the art as the tetrazine ligation reaction.

[0749]

[0305] In certain embodiments, when the conjugate is formed through a tetrazine inverse electron demand Diels - Alder ligation reaction, the divalent residue of the reactive group (e.g., RL) includes a fused bicyclic ring having at least two adjacent nitrogen atoms within the ring. In certain embodiments, when the conjugate is formed through a tetrazine inverse electron demand Diels - Alder ligation reaction, the divalent residue of the reactive group (e.g., RL) is

[0750]

Chem.

[0751]

[0306] When the conjugate of formula (II)-(IIH), (IV), (IVA), or (IVB) is formed by an inverse electron demand ligation reaction, the conjugate includes both regioisomers. In certain embodiments, the conjugate of formula (II)-(IIH), (IV), (IVA), or (IVB) is a mixture of regioisomers formed from an inverse electron demand ligation reaction.

[0752]

[0307] Thiol reaction

[0753]

Chem.

[0754]

[0308] Advantageously, compounds containing a terminal thiol group or a suitable electrophilic group or disulfide-forming group promote a selective and efficient reaction with a second compound containing a complementary electrophilic group or disulfide-forming group or thiol group. These reactions are thought to be selective with little to no cross-reactivity with functional groups within biomolecules. In some embodiments, the thiol reaction does not include the reaction of a maleimide group.

[0755]

[0309] In certain embodiments, when the conjugate is formed through a thiol-maleimide reaction, the divalent residue of the reactive group is

[0756]

Chem.

[0757]

Chem.

[0758]

Chem.

[0759]

Chem.

[0760]

Chem.

[0761]

[0310] Carbonyl-oxyamine reaction

[0762]

Chem.

[0763]

[0311] Advantageously, compounds containing a terminal carbonyl or oxyamine group facilitate a selective and efficient reaction with a second compound containing an oxyamine or carbonyl group. The carbonyl and oxyamine are thought to react to form an oxime bond. The reaction is thought to be specific with little to no cross-reactivity with functional groups within biomolecules.

[0764]

[0312] In certain embodiments, when the conjugate of formula (II)-(IIH), (IV), (IVA), or (IVB) is formed through an oxime conjugation reaction, the divalent residue of the reactive group comprises the divalent residue of a non-natural amino acid. In certain embodiments, when the conjugate of formula (II)-(IIH), (IV), (IVA), or (IVB) is formed through an oxime conjugation reaction, the divalent residue of the reactive group (e.g., RL) is

[0765]

Chem.

[0766]

Chem.

[0767]

[0313] Thiol-N-hydroxysuccinimide reaction

[0768]

Chem.

[0769]

[0314] Advantageously, a compound comprising a terminal thiol-N-hydroxysuccinimide promotes the reaction with a second compound comprising an amine group, such as glutamine, to form an amide.

[0770]

[0315] In certain embodiments, when the conjugates of formulas (II)-(IIH), (IV), (IVA), or (IVB) are formed through a thiol-N-hydroxysuccinimide conjugation reaction, the divalent residue of the reactive group comprises the divalent residue of an amino acid, such as glutamine. In certain embodiments, when the conjugates of formulas (II)-(IIH), (IV), (IVA), or (IVB) are formed through a thiol-N-hydroxysuccinimide conjugation reaction, the divalent residue of the reactive group (e.g., RL) is

[0771]

Chemical formula

[0772]

[0316] Other reactions

[0773]

[0317] Other suitable conjugation reactions are described in the literature. See, for example, Lang, K. and Chin, J. 2014, Bioorthogonal Reactions for Labeling Proteins, ACS Chem Biol 9, 16-20; Paterson, D.M. et al. 2014, Finding the Right (Bioorthogonal) Chemistry, ACS Chem Biol 9, 592-605; King, M. and Wagner, A. 2014, Developments in the Field of Bioorthogonal Bond Forming Reactions - Past and Present Trends, Bioconjugate Chem., 2014, 25(5), 825-839; and Ramil, C.P. and Lin, Q., 2013, Bioorthogonal chemistry: strategies and recent developments, Chem Commun 49, 11007-11022.

[0774]

[0318] Release reaction

[0775]

[0319] A release reaction is a reaction that acts to release a biologically active moiety of a compound or conjugate described herein in vivo and / or in vitro from the compound or conjugate. In certain embodiments, the biologically active moiety released is a compound (e.g., a cytotoxic agent) described elsewhere herein, or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof. An example of a release reaction is an intramolecular reaction between an eliminator group and a release-inducing group of a compound or conjugate described herein that releases the biologically active moiety of the compound or conjugate described herein. The eliminator group itself can be transformed into two reactive components as exemplified in these reactions, and X is a drug having a heteroatom for bonding that is nitrogen or oxygen. Exemplary release reactions are shown in the following scheme:

[0776] [Chem.]

[0777]

[0320] Water-soluble polymer In certain embodiments, the compounds or conjugates described herein comprise one or more water-soluble polymers. A wide variety of polymeric polymers and other molecules can be linked to the polypeptides described herein to modulate the biological properties of the polypeptides and / or to confer new biological properties on the polypeptides. These polymeric polymers can be linked to the polypeptide via naturally encoded amino acids, via non-naturally encoded amino acids, or via any functional substituent of a natural or modified amino acid, or via any substituent or functional group added to a natural or modified amino acid. The molecular weight of the polymer can include a wide range including, but not limited to, between about 100 Da and about 100,000 Da or more.

[0778]

[0321] The polymer selected can be water-soluble such that the protein to which it is attached does not precipitate in an aqueous environment such as a physiological environment. The polymer can be branched or unbranched. In certain embodiments, the polymer will be pharmaceutically acceptable for therapeutic use of the final product preparation.

[0779]

[0322] In certain embodiments, the ratio of polyethylene glycol molecules to polypeptide molecules will vary as their concentrations in the reaction mixture. Generally, the optimal ratio (in terms of the efficiency of the reaction in the sense that there is a minimum amount of excess unreacted protein or polymer present) can be determined by the number of available reactive groups, depending on the molecular weight of the polyethylene glycol selected. With respect to molecular weight, typically, the higher the molecular weight of the polymer, the fewer the number of polymer molecules that can be attached to the protein. Similarly, when optimizing these parameters, the branching of the polymer should be considered. Generally, the higher the molecular weight (or the more branched), the higher the polymer:protein ratio.

[0780]

[0323] The water-soluble polymer can be of any structural form including, but not limited to, linear, forked, or branched. Typically, the water-soluble polymer is a poly(alkylene glycol) such as poly(ethylene glycol) (PEG), although other water-soluble polymers can also be employed. As an example, certain embodiments will be described using PEG.

[0781]

[0324] PEG is a well-known water-soluble polymer that is commercially available or can be prepared by ring-opening polymerization of ethylene oxide according to methods well known in the art (Sandler and Karo, Polymer Synthesis, Academic Press, New York, Volume 3, pages 138 - 161). The term "PEG" is widely used to encompass any polyethylene glycol molecule, regardless of size or modification of the ends of the PEG, and can be represented as being linked to the polypeptide by the formula: X’O-(CH2CH2O) n -CH2CH2-Y’ (wherein n is an integer selected from 2 to 10,000, X’ is hydrogen or, among other things, a terminal modification containing C 1~4 alkyl, and Y’ is a point of attachment to the polypeptide).

[0782]

[0325] In some cases, the PEG terminates at one end with a hydrogen or a methoxy, i.e., X’ is hydrogen or CH3 (also known as "methoxy PEG"). Alternatively, the PEG can terminate with a PEG-reactive group, thereby forming a bifunctional polymer. Typical PEG-reactive groups include functional groups found in the 20 common amino acids (but not limited thereto), such as maleimide groups, activated carbonates (including but not limited to p-nitrophenyl esters), activated esters (including but not limited to N-hydroxysuccinimide, p-nitrophenyl esters, and aldehydes), and functional groups that react specifically with complementary functional groups present in non-naturally encoded amino acids (but not limited thereto, including azide groups and / or alkyne groups) that are inert to the 20 common amino acids. It is noted that the other end of the PEG represented by the above formula by Y’ attaches directly or indirectly to the polypeptide via a naturally occurring amino acid or a non-naturally encoded amino acid. For example, Y’ can be an amide bond, a carbamate bond, or a urea bond with an amine group of the polypeptide (including but not limited to the ε-amine or N-terminus of lysine). Alternatively, Y’ can be a maleimide bond with a thiol group (including but not limited to the thiol group of cysteine). Alternatively, Y’ can be a bond with a residue that is not generally available via the 20 common amino acids. For example, an azide group on the PEG can react with an alkyne group on the polypeptide to form a Huisgen [3+2] cycloaddition product. Alternatively, an alkyne group on the PEG can react with an azide group present in a non-naturally encoded amino acid such as a modified amino acid described herein to form a similar product. In some embodiments, strong nucleophiles (including but not limited to hydrazine, hydrazide, hydroxylamine, or semicarbazide) react with aldehyde or ketone groups present in non-naturally encoded amino acids to form, where applicable, hydrazones, oximes, or semicarbazones, which can be further reduced in some cases by treatment with an appropriate reducing agent.Alternatively, a strong nucleophilic reagent can be incorporated into a polypeptide via a non-naturally encoded amino acid and can be used to preferentially react with a ketone or aldehyde group present in a water-soluble polymer.

[0783]

[0326] Not limited thereto, any molecular weight of PEG including a desired about 100 Daltons (Da) to 100,000 Da or more (not limited thereto, in certain embodiments, including 0.1 to 50 kDa or 10 to 40 kDa) can be used as actually desired. Not limited thereto, branched-chain PEGs containing PEG molecules having a molecular weight (MW) in the range of 1 to 100 kDa (not limited thereto, including 1 to 50 kDa or 5 to 20 kDa) for each chain can also be used. A wide range of PEG molecules are described in the catalogs of Shearwater Polymers, Inc. and Nektar Therapeutics, each of which is incorporated herein by reference.

[0784]

[0327] Generally, at least one end of the PEG molecule is available for reaction with the remainder of the compounds of formulas (I)-(IVB). For example, a PEG derivative having an alkyne moiety and an azide moiety for reaction with an amino acid side chain can be used to attach PEG to a non-naturally encoded amino acid described herein. When the non-naturally encoded amino acid contains an azide, the PEG typically contains either an alkyne moiety for formation of a [3+2] cycloaddition product or an activated PEG species (i.e., an ester, a carbonate) containing a phosphine group for formation of an amide bond. Alternatively, when the non-naturally encoded amino acid contains an alkyne, the PEG typically contains an azide moiety for formation of a [3+2] Huisgen cycloaddition product. When the non-naturally encoded amino acid contains a carbonyl group, the PEG typically contains a nucleophile (including, but not limited to, a hydrazide, hydrazine, hydroxylamine, or semicarbazide functional group) for formation of the corresponding hydrazone bond, oxime bond, and semicarbazone bond, respectively. In other alternative forms, the reverse orientation of the reactive groups described herein can be used (i.e., the azide moiety of the non-naturally encoded amino acid can be reacted with a PEG derivative containing an alkyne).

[0785]

[0328] In some embodiments, the polypeptide variant having the PEG derivative contains a chemical functional group that is reactive with a chemical functional group present on the side chain of a non-naturally encoded amino acid.

[0786]

[0329] In certain embodiments, the water-soluble polymer is an azide-containing polymer or an acetylene-containing polymer comprising a water-soluble polymer backbone having an average molecular weight of from about 800 Da to about 100,000 Da. The polymer backbone of the water-soluble polymer can be poly(ethylene glycol). However, without limitation, a wide variety of water-soluble polymers including poly(ethylene glycol) as well as other related polymers including poly(dextran) and poly(propylene glycol) are suitable for use, and the use of the term "PEG" or "poly(ethylene glycol)" is intended to encompass and include all such molecules. The term "PEG" further includes poly(ethylene glycol) in any of its forms including, without limitation, bifunctional PEG, multi-arm PEG, derivatized PEG, dendritic PEG, branched PEG, pendant PEG (i.e., PEG or related polymer having one or more functional groups pendant from the polymer backbone), or PEG having a degradable bond therein.

[0787]

[0330] The polymer backbone can be linear or branched. Branched polymer backbones are generally known in the art. Typically, a branched polymer has a central branched core portion and a plurality of linear polymer chains linked to the central branched core. PEG is commonly used in a branched form that can be prepared by adding ethylene oxide to various polyols such as glycerin, glycerin oligomers, pentaerythritol, and sorbitol. The central branched portion can also be derived from some amino acids such as lysine. Branched poly(ethylene glycol) is R-(-PEG-OH) m(In the formula, R is derived from a core moiety such as glycerin, glycerin oligomer, or pentaerythritol, and m represents the number of arms), and can be represented in a general form. Multi-arm PEG molecules, for example, U.S. Patent Nos. 5,932,462; 5,643,575; 5,229,490; and 4,289,872, each of which is incorporated herein by reference in its entirety; U.S. Patent Application Publication No. 2003 / 0143596; and International Publication Nos. 96 / 21469 and 93 / 21259, can also be used as the polymer backbone.

[0788]

[0331] Branched PEG can also be in the form of forked PEG represented by PEG(-Y’’CHZ2) n (wherein Y’’ is a linking group and Z is an activated terminal group linked to CH by a chain of atoms of a predetermined length).

[0789]

[0332] Another branched form, pendant PEG, has a PEG-reactive group such as carboxyl along the PEG backbone rather than at the end of the PEG chain.

[0790]

[0333] In addition to these forms of PEG, the polymer can also be prepared with labile or degradable bonds within the backbone. For example, PEG can be prepared with an ester bond within a polymer backbone that is susceptible to hydrolysis. As shown herein, this hydrolysis cleaves the polymer into low molecular weight fragments: -PEG-CO2-PEG- + H2O → PEG-CO2H + HO-PEG-. The term "poly(ethylene glycol)" or "PEG" represents, but is not limited to, all forms known in the art including those disclosed herein, or is understood by those skilled in the art to include these.

[0791]

[0334] Many other polymers are also suitable for use. In some embodiments, water-soluble polymer backbones having from 2 to about 300 termini are particularly suitable. Examples of suitable polymers include, but are not limited to, other poly(alkylene glycols) such as poly(propylene glycol) (“PPG”), copolymers thereof (including, but not limited to, copolymers of ethylene glycol and propylene glycol), terpolymers thereof, mixtures thereof, and the like. The molecular weight of each chain of the polymer backbone can vary, but typically ranges from about 800 Da to about 100,000 Da, and often from about 6,000 Da to about 80,000 Da.

[0792]

[0335] Those skilled in the art will recognize that the foregoing list of substantially water-soluble backbones is in no way exhaustive and is merely illustrative, and that all polymer materials having the qualities described herein are contemplated to be suitable for use.

[0793]

[0336] In some embodiments, the polymer derivative is “multifunctional,” which means that the polymer backbone has at least 2 termini, and perhaps up to about 300 termini, that are functionalized or activated with functional groups. Multifunctional polymer derivatives include, but are not limited to, linear polymers having two termini that are attached to functional groups that may be the same or different at each terminus.

[0794]

[0337] Compositions and Uses Pharmaceutical Compositions and Methods of Administration The conjugates provided herein can be formulated into pharmaceutical compositions using methods available in the art and the methods disclosed herein. Any of the conjugates provided herein can be provided in a suitable pharmaceutical composition and administered by a suitable route of administration.

[0795]

[0338] The methods provided herein include administering a pharmaceutical composition comprising at least one conjugate provided herein and one or more pharmaceutically acceptable carriers of compatibility. In this context, the term "pharmaceutically acceptable" means approved by a regulatory authority of the federal or state government or listed in the United States Pharmacopeia or other generally recognized pharmacopeias for use in animals and, in certain embodiments, in humans. The term "carrier" includes diluents, adjuvants (e.g., Freund's adjuvant (complete and incomplete)), excipients, or vehicles with which a therapeutic agent can be administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water can be used as a carrier when the pharmaceutical composition is administered intravenously. Physiological saline as well as aqueous dextrose and glycerol solutions can also be employed especially as liquid carriers for injectable solutions. Examples of suitable pharmaceutical carriers are described in Martin, E.W., Remington’s Pharmaceutical Sciences.

[0796]

[0339] In clinical practice, the pharmaceutical compositions or conjugates provided herein can be administered by any route known in the art. Exemplary routes of administration include, but are not limited to, oral, inhalation, intraarterial, intradermal, intramuscular, intraperitoneal, intravenous, intranasal, parenteral, pulmonary, and subcutaneous routes. In some embodiments, the pharmaceutical compositions or conjugates provided herein are administered orally. In some embodiments, the pharmaceutical compositions or conjugates provided herein are administered parenterally.

[0797] The composition for parenteral administration can be an emulsion or a sterile solution. The parenteral composition can include, for example, propylene glycol, polyethylene glycol, vegetable oil, and organic esters for injection (e.g., ethyl oleate). These compositions can also contain wetting agents, isotonic agents, emulsifying agents, dispersing agents, and stabilizing agents. Sterilization can be carried out in several ways, for example, using a bacterial filter, through radiation, or through heating. The parenteral composition can also be prepared in the form of a sterile solid composition that can be dissolved in sterile water or any other sterile medium for injection at the time of use.

[0798]

[0341] In some embodiments, the compositions provided herein are pharmaceutical compositions or single unit dosage forms. The pharmaceutical compositions and single unit dosage forms provided herein contain a prophylactically effective amount or a therapeutically effective amount of one or more prophylactic conjugates or therapeutic conjugates.

[0799]

[0342] The pharmaceutical composition may contain one or more pharmaceutical excipients. Any suitable pharmaceutical excipient may be used, and a person skilled in the art can select a suitable pharmaceutical excipient. Non-limiting examples of suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, nonfat dry milk, glycerin, propylene glycol, water, ethanol, and the like. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on various factors well known in the art, including but not limited to the method by which the dosage form is to be administered to a subject and the particular conjugate in the dosage form. The composition or single unit dosage form may also contain, if desired, trace amounts of a wetting or emulsifying agent, or a pH buffering agent. Accordingly, the pharmaceutical excipients provided herein are intended to be exemplary and not limiting. Additional pharmaceutical excipients include, for example, those described in Handbook of Pharmaceutical Excipients, Rowe et al. (eds.) 6th Edition (2009), which is hereby incorporated by reference in its entirety.

[0800]

[0343] In some embodiments, the pharmaceutical composition contains an antifoaming agent. Any suitable antifoaming agent may be used. In some aspects, the antifoaming agent is selected from alcohols, ethers, oils, waxes, silicones, surfactants, and combinations thereof. In some aspects, the antifoaming agent is selected from mineral oil, vegetable oil, ethylene bisstearamide, paraffin wax, ester wax, fatty alcohol wax, long-chain fatty alcohol, fatty acid soap, fatty acid ester, silicone glycol, fluorosilicone, polyethylene glycol - polypropylene glycol copolymer, polydimethylsiloxane - silicon dioxide, ether, octyl alcohol, capryl alcohol, sorbitan trioleate, ethyl alcohol, 2-ethyl-hexanol, dimethicone, oleyl alcohol, simethicone, and combinations thereof.

[0801]

[0344] In some embodiments, the pharmaceutical composition comprises a co-solvent. Exemplary examples of co-solvents include ethanol, poly(ethylene) glycol, butylene glycol, dimethylacetamide, glycerin, and propylene glycol.

[0802]

[0345] In some embodiments, the pharmaceutical composition comprises a buffering agent. Exemplary examples of buffering agents include acetates, borates, carbonates, lactates, malates, phosphates, citrates, hydroxides, diethanolamine, monoethanolamine, glycine, methionine, guar gum, and sodium glutamate.

[0803]

[0346] In some embodiments, the pharmaceutical composition comprises a carrier or filler. Exemplary examples of carriers or fillers include lactose, maltodextrin, mannitol, sorbitol, chitosan, stearic acid, xanthan gum, and guar gum.

[0804]

[0347] In some embodiments, the pharmaceutical composition comprises a surfactant. Exemplary examples of surfactants include d-α tocopherol, benzalkonium chloride, benzethonium chloride, cetrimide, cetylpyridinium chloride, doxart sodium, glyceryl behenate, glyceryl monooleate, lauric acid, macrogol 15 hydroxystearate, myristyl alcohol, phospholipids, polyoxyethylene alkyl ether, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene stearate, polyoxyl glyceride, sodium lauryl sulfate, sorbitan ester, and vitamin E polyethylene (glycol) succinate.

[0805]

[0348] In some embodiments, the pharmaceutical composition comprises an anti-caking agent. Exemplary examples of anti-caking agents include (tertiary) calcium phosphate, hydroxymethylcellulose, hydroxypropylcellulose, and magnesium oxide.

[0806] Other excipients that can be used in pharmaceutical compositions include, for example, albumin, antioxidants, antibacterial agents, antifungal agents, biocompatible polymers, chelating agents, controlled release agents, diluents, dispersants, solubilizing agents, emulsifiers, gelling agents, ointment bases, penetration enhancers, preservatives, solubilizers, solvents, stabilizers, and sugars. Specific examples of each of these agents are described, for example, in Handbook of Pharmaceutical Excipients, Rowe et al. (eds.), 6th Edition (2009), The Pharmaceutical Press, which is hereby incorporated by reference in its entirety.

[0807]

[0350] In some embodiments, the pharmaceutical composition includes a solvent. In some aspects, the solvent is a saline solution such as sterile isotonic saline or a dextrose solution. In some aspects, the solvent is water for injection.

[0808]

[0351] In some embodiments, the pharmaceutical composition is in particulate form such as microparticles or nanoparticles. The microparticles and nanoparticles can be formed from any suitable material such as a polymer or lipid. In some aspects, the microparticles or nanoparticles are micelles, liposomes, or polymersomes.

[0809]

[0352] In some embodiments, since water can promote the degradation of some antibodies or antigen-binding fragments thereof, anhydrous pharmaceutical compositions and dosage forms containing conjugates are further provided herein.

[0810]

[0353] The anhydrous pharmaceutical compositions and dosage forms provided herein can be prepared using anhydrous or low-moisture-containing components and low-moisture or low-humidity conditions. When substantial contact with moisture and / or humidity during manufacture, packaging, and / or storage is anticipated, pharmaceutical compositions and dosage forms containing lactose and at least one active ingredient containing a primary or secondary amine can be anhydrous.

[0811]

[0354] The anhydrous pharmaceutical compositions can be prepared and stored such that their anhydrous nature is maintained. Thus, the anhydrous compositions can be packaged using materials known to prevent exposure to water so that they can be included in suitable dosing kits. Examples of suitable packaging include, but are not limited to, sealed foils, plastics, unit dose containers (e.g., vials), blister packs, and strip packs.

[0812]

[0355] The lactose-free compositions provided herein are well known in the art and can include excipients such as those listed in the United States Pharmacopeia (USP) SP(XXI) / NF(XVI). Generally, lactose-free compositions include a pharmaceutically compatible and pharmaceutically acceptable amount of an active ingredient, a binder / filler, and a lubricant. Exemplary lactose-free dosage forms include an active ingredient, microcrystalline cellulose, pregelatinized starch, and magnesium stearate.

[0813]

[0356] Pharmaceutical compositions and dosage forms are also provided that include one or more excipients that decrease the rate at which the conjugate degrades. Such excipients are also referred to herein as "stabilizers" and include, but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers.

[0814]

[0357] Parenteral Dosage Forms In certain embodiments, parenteral dosage forms are provided. Parenteral dosage forms can be administered to a subject by a variety of routes including, but not limited to, subcutaneous, intravenous (including bolus injection), intramuscular, and intraarterial. Since such administrations typically bypass the subject's natural defenses against contaminants, parenteral dosage forms are typically sterile or can be sterilized prior to administration to the subject. Examples of parenteral dosage forms include, but are not limited to, solutions that can be immediately injected, dry products that can be immediately dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions that can be immediately injected, and emulsions.

[0815]

[0358] Suitable vehicles that can be used to provide a parenteral dosage form are well known to those of skill in the art. Examples include, but are not limited to, Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[0816]

[0359] Excipients that increase the solubility of one or more of the antibodies disclosed herein can also be incorporated into the parenteral dosage form.

[0817]

[0360] Dosage and Unit Dosage Form In human therapeutics, the physician will determine the most appropriate pharmacology, in accordance with prophylactic or therapeutic treatment, and in accordance with age, body weight, condition, and other factors specific to the subject being treated.

[0818]

[0361] In certain embodiments, the compositions provided herein are pharmaceutical compositions or single unit dosage forms. The pharmaceutical compositions and single unit dosage forms provided herein contain a prophylactically effective amount or a therapeutically effective amount of one or more prophylactic antibodies or therapeutic antibodies or antigen-binding fragments thereof.

[0819]

[0362] The amount of the conjugate or composition effective in the prevention or treatment of a disorder or one or more of its symptoms will vary depending on the nature and severity of the disease or condition and the route by which the conjugate is administered. The frequency and dosage will also vary according to factors specific to each subject that depend on the particular treatment being administered (e.g., therapeutic or prophylactic agent), the severity of the disorder, disease, or condition, the route of administration, and the age, weight, response, and past medical history of the subject. An effective dosage can be extrapolated from a dose-response curve derived from in vitro or animal model test systems.

[0820]

[0363] In certain embodiments, exemplary dosages of the conjugate or composition include antibody amounts in milligrams or micrograms per kilogram of subject or sample weight (e.g., from about 10 micrograms per kilogram to about 50 milligrams per kilogram, from about 100 micrograms per kilogram to about 25 milligrams per kilogram, or from about 100 micrograms per kilogram to about 10 milligrams per kilogram).

[0821]

[0364] The dosage can be administered according to a suitable schedule. As will be apparent to those skilled in the art, in some cases it may be necessary to use dosages of the antibody conjugate outside the ranges disclosed herein. Further, it is noted that the clinician or treating physician will know how and when to interrupt, adjust, or terminate the treatment in accordance with the response of the subject.

[0822] As will be readily appreciated by those skilled in the art, different therapeutically effective amounts may be applicable to different diseases and conditions. Similarly, an amount that is sufficient to prevent, manage, treat, or improve such a disorder but insufficient to cause an adverse effect associated with the antibodies or antigen-binding fragments thereof provided herein, or an amount sufficient to reduce an adverse effect, is also encompassed by the dosage and dosing frequency schedules described herein. Further, when multiple doses of the compositions provided herein are administered to a subject, not all of the doses need be the same. For example, the dose administered to a subject can be increased to improve the prophylactic or therapeutic effect of the composition or decreased to reduce one or more side effects experienced by a particular subject.

[0823]

[0366] In certain embodiments, treatment or prophylaxis can be initiated with one or more loading doses of the conjugates or compositions provided herein, followed by one or more maintenance doses.

[0824]

[0367] In certain embodiments, a dose of the conjugates or compositions provided herein can be administered to achieve a steady-state concentration of the conjugate in the blood or serum of a subject. The steady-state concentration can be determined by measurements according to techniques available to those skilled in the art or can be based on physical characteristics of the subject such as height, weight, and age.

[0825]

[0368] Therapeutic Use For therapeutic use, the conjugate is administered to a mammal, in certain embodiments, a human, in a pharmaceutically acceptable dosage form, such as dosage forms known in the art and discussed herein. For example, the conjugates of the present disclosure can be administered to a human intravenously as a bolus or by continuous infusion over a period of time, by intramuscular, intraperitoneal, intrathecal, subcutaneous, intraarterial, intra-articular synovial sac, intramedullary, or intratumoral routes. The conjugate can also be appropriately administered by peritumoral, intralesional, or perilesional routes to exert local and systemic therapeutic effects. The intraperitoneal route can be particularly useful, for example, in the treatment of ovarian tumors.

[0826]

[0369] The conjugates provided herein can be useful for the treatment of any disease or condition described herein (e.g., inflammatory and / or proliferative diseases or conditions). In some embodiments, the disease or condition is a disease or condition that can be diagnosed by overexpression of an antigen. In some embodiments, the disease or condition is a disease or condition that can benefit from treatment with a macromolecule. In some embodiments, the disease or condition is abnormal cell proliferation.

[0827]

[0370] In some embodiments, the disease or condition is cancer. In certain embodiments, the cancer is small cell lung cancer, non-small cell lung cancer, ovarian cancer, platinum-resistant ovarian cancer, ovarian adenocarcinoma, endometrial cancer, breast cancer, breast cancer overexpressing HER2, triple-negative breast cancer, lymphoma, large cell lymphoma, diffuse mixed histiocytic and lymphocytic lymphoma, follicular B cell lymphoma, colon cancer, colorectal cancer, colon adenocarcinoma, colorectal adenocarcinoma, melanoma, prostate cancer, or multiple myeloma.

[0828]

[0371] Diagnostic Use In some embodiments, the conjugates provided herein are used for diagnostic purposes. These assays can be useful, for example, for diagnosing and / or prognosticating diseases such as cancer.

[0829]

[0372] In some diagnostic and prognostic uses or embodiments, the conjugate may be labeled with a detectable moiety. Suitable detectable moieties include, but are not limited to, radioisotopes, fluorescent labels, and enzyme-substrate labels. In another embodiment, the conjugate need not be labeled, and the presence of the conjugate can be detected using a labeled antibody or an antigen-binding fragment thereof that specifically binds to the conjugate.

[0830]

[0373] Kit In some embodiments, the conjugates provided herein are provided in the form of a kit (i.e., a packaged combination of a predetermined amount of reagents and instructions for performing a procedure). In some embodiments, the procedure is a diagnostic assay. In certain embodiments, the procedure is a therapeutic procedure.

[0831]

[0374] In some embodiments, the kit further includes a solvent for reconstituting the conjugate. In some embodiments, the conjugate is provided in the form of a pharmaceutical composition.

[0832]

[0375] In some embodiments, the kit can include the conjugates or compositions provided herein, an optional second agent or composition, and instructions for providing healthcare providers with information regarding use for treating a disorder. The instructions can be provided in printed form, or in the form of an electronic medium such as a floppy disk, CD, or DVD, or in the form of a website address from which such instructions can be obtained. The unit dosage of the conjugates or compositions provided herein, or the second agent or composition, can include an amount that, when administered to a subject, can maintain a therapeutically effective plasma level or prophylactically effective plasma level of the compound or composition in the subject for at least one day. In some embodiments, the compound or composition can be included as a sterile aqueous pharmaceutical composition or a dry powder (e.g., lyophilized) composition.

[0833]

[0376] In some embodiments, appropriate packaging is provided. As used herein, "packaging" includes solid matrices or materials that are customarily used in the art and that can hold, within certain limits, the compounds provided herein and / or a second agent suitable for administration to a subject. Such materials include glass and plastic (e.g., polyethylene, polypropylene, and polycarbonate) bottles, vials, paper, plastic, plastic foil laminate envelopes, etc. When electron beam sterilization techniques are employed, the packaging should have a density low enough to allow sterilization of the contents.

[0834]

[0377] Preparation and synthesis procedures Conjugation Conjugates can be prepared by standard techniques. In certain embodiments, a polymer is contacted with a compound of formula (I)-(IH), (III), (IIIA), or (IIIB) under conditions suitable to form a bond from the polymer to formula (I)-(IH), (III), (IIIA), or (IIIB) to form a conjugate of, for example, formula (II)-(IIH), (IV), (IVA), or (IVB). In certain embodiments, a polymer is contacted with a linker precursor under conditions suitable to form a bond from the polymer to the linker. The resulting polymer-linker is contacted with a compound or drug moiety under conditions suitable to form a bond from the polymer-linker to the compound or drug moiety to form a conjugate. In certain embodiments, a compound or drug moiety is contacted with a linker precursor under conditions suitable to form a bond from the compound or drug moiety to the linker. The resulting compound-linker or drug moiety-linker is contacted with a polymer under conditions suitable to form a bond from the compound-linker or drug moiety-linker to the polymer to form a conjugate. For example, in certain embodiments, the second compound comprises tetrazine; and the RG comprises a strained alkene. In some embodiments, the RG is

[0835] [Chemistry] It is. In certain embodiments, the second compound contains an azide; RG contains an alkyne. In some embodiments, RG is

[0836] [Chemistry] It is. In certain embodiments, the second compound contains an alkyne; RG contains an azide. In certain embodiments, the second compound contains a strained alkene; RG contains a tetrazine. In certain embodiments, the second compound contains a thiol; RG contains a maleimide. In some embodiments, RG is

[0837] [Chemistry] It is. In certain embodiments, the second compound contains a maleimide; RG contains a thiol. In some embodiments, the second compound is

[0838] [Chemistry] including. In certain embodiments, the second compound contains a carbonyl; RG contains an oxyamine. In some embodiments, RG is

[0839] [Chemistry] It is. In some embodiments, the second compound is

[0840] [Chemistry] including. In certain embodiments, the second compound contains an oxyamine; RG contains a carbonyl. In certain embodiments, RG is

[0841] [Chemical formula] is. In some embodiments, RG is

[0842] [Chemical formula] is. In some embodiments, RG is

[0843] [Chemical formula] is. In certain embodiments, the second compound is

[0844] [Chemical formula] comprises. In certain embodiments, the second compound is a polypeptide. In certain embodiments, the second compound is an antibody. In certain embodiments, the second compound is an antibody chain. Linkers suitable for preparing the conjugate are disclosed herein, and exemplary conditions for conjugation are described in the following examples. [Examples]

[0845]

[0378] The compounds provided herein can be prepared, isolated, or obtained by any method apparent to those skilled in the art. The compounds provided herein can be prepared according to the exemplary preparation schemes provided below. Reaction conditions, steps, and reactants not provided in the exemplary preparation schemes will be apparent to those skilled in the art and will be known to those skilled in the art. As used herein, the symbols and conventions used in these processes, schemes, and examples are consistent with those used in modern scientific literature, such as the Journal of the American Chemical Society or the Journal of Biological Chemistry, whether or not specific abbreviations are explicitly defined. Specifically, but not limited to, the following abbreviations may be used in the examples and throughout the specification: g (gram); mg (milligram); mL (milliliter); μL (microliter); mM (millimolar concentration); μM (micromolar concentration); Hz (hertz); MHz (megahertz); mmol (millimole); h, hr, or hrs (hour); min (minute); MS (mass spectrometry); ESI (electrospray ionization); LCMS (liquid chromatography - mass spectrometry); TLC (thin - layer chromatography); HPLC (high - performance liquid chromatography); rt (room temperature); atm (atmosphere); cald (calculated); equiv (equivalent); CDCl3 (deuterochloroform); DBCO (dibenzocyclooctyne - amine); DCE (dichloroethane); DCM (dichloromethane); DIPEA (diisopropylethylamine); DMSO (dimethyl sulfoxide); DMSO - d6 (deuterated dimethyl sulfoxide); EtOAc (ethyl acetate); EtOH (ethanol); MeCN (acetonitrile); MeOH (methanol); RB (round - bottom flask); TFA (trifluoroacetic acid); THF (tetrahydrofuran); DMF (dimethylformamide); and BOC (t - butyloxycarbonyl).

[0846]

[0379] For all of the following examples, standard post-treatment and purification methods known to those skilled in the art can be utilized. Unless otherwise indicated, all temperatures are expressed in °C (degrees Celsius). Unless otherwise noted, all reactions are carried out at room temperature. The synthetic methodologies exemplified in this specification are intended to illustrate the applicable chemistry through the use of specific examples and do not indicate the scope of the present disclosure.

[0847]

[0380] Unless otherwise indicated, all anhydrous solvents were commercially obtained and stored in Sure-Seal bottles under nitrogen. All other reagents and solvents were purchased as the highest available grade and used without further purification. NMR spectra were recorded on an Avance II HD (500 MHz) spectrometer equipped with a 5 mm Prodigy H / F-BBO cryoprobe and a BCU-I temperature controller. Chemical shifts (δ) were reported in parts per million (ppm) relative to tetramethylsilane at δ 0.00, and coupling constants (J) were reported in Hz. Low-resolution mass spectral data were acquired on an Agilent G6125B spectrometer interfaced with an Agilent 1260 high-performance liquid chromatography instrument for LC-MS. The products were purified by RP-HPLC using a Shimadzu LC equipped with a CTC IFC, a Phenomenex Gemini NX 5μ, C18, 110 Å, 150 × 50 mm reversed-phase column, with a linear gradient of B (CH3CN) in mobile phase A (water + 0.1% TFA) at a flow rate of 50 mL / min. Analytical HPLC was performed on a Waters 2695 instrument. For analytical HPLC, the stationary phase used was a Phenomenex Gemini NX 5μ, C18, 110 Å, 150 × 4.6 mm RP column. The products were eluted at a flow rate of 1.0 mL / min with either an acidic linear gradient (referred to as gradient A) of B (CH3CN + 0.05% TFA; 5% - 95% over 20 minutes) in mobile phase A (0.05% aqueous TFA). Preparative HPLC purification was carried out on a Shimadzu LC equipped with a CTC IFC. All other preparative normal-phase purifications were performed by standard flash silica gel chromatography using an ISCO flash system.

[0848]

[0381] Example 1. Synthesis of DBCO-nnAA-PEG13-AAN-Exatecan (Compound 1):

[0849]

[0382]

[0850]

Chem.

[0851]

[0383] A suspension of exatecan mesylate 1 (MsOH salt, 150 mg, 0.28 mmol) in anhydrous DMF (3 mL) was added with Fmoc-AAN(Trt)-OH 2 (250 mg, 0.33 mmol), EDC (65 mg, 0.34 mmol), HOAt (46 mg, 0.34 mmol), and DIPEA (53 μL) at room temperature. When the reaction mixture was stirred at RT for 1 h, LCMS indicated the desired product, then 0.3 mL of piperidine was added. The mixture was stirred for 5 min and then added to a 1 / 1 mixture of hexane / diethyl ether (45 mL). The precipitate was collected by centrifugation and the solvent was removed by decantation to obtain Compound 2b. The residue 2b was dissolved in 3 mL of TFA and the mixture was stirred at RT for 10 min. Then, TFA was removed under reduced pressure and the crude mixture was purified by reverse-phase HPLC. Freeze-drying of the pure fractions gave Compound 3 as the TFA salt (63 mg); LCMS m / z (ESI + ): Calculated for C 34 H 38 FN7O8, 691.28; found 692.4 (M+H).

[0852]

[0384]

[0853]

Chem.

[0854]

[0385] Synthesis of Compound 7: Compound 5 (1.4 g, 3.34 mmol) was dissolved in DMF (10 mL). To the clear solution, HATU (1.2 g, 3.34 mmol) and DIPEA (861 mg, 6.68 mmol) were added. The solution was stirred at room temperature for about 30 seconds, and subsequently DBCO-amine 7 (922 mg, 3.34 mmol) in DMF (2 mL) was added. After the mixture was stirred at room temperature for 20 minutes, diethylamine (2 mL) was added and stirring was continued for another 30 minutes. LCMS indicated the completion of the reaction. The reaction solution was concentrated under reduced pressure and purified by reverse-phase HPLC to obtain Compound 7 (790 mg); LCMS m / z (ESI + ): Calculated for C 29 H 33 N3O2, 455.26; Found 456.4 (M+H).

[0855]

[0386] Synthesis of Compound 4: A solution of Compound 8 (3 g, 2.9 mmol), Compound 7 (780 mg, 1.37 mmol), and DIPEA (353 mg, 2.74 mmol) in DMF (20 mL) was stirred for 20 minutes. LCMS indicated the completion of the reaction. The reaction mixture was directly purified by reverse-phase HPLC to obtain Compound 4 (1.52 g); LCMS m / z (ESI + ): Calculated for C 65 H 88 F5N3O 18 1293.60; Found 1293.7 (M+H).

[0856]

[0387]

[0857]

Chemical Structure

[0858] To a solution of compound 4 (770 mg, 0.6 mmol) in anhydrous DMF (3 mL) was added compound 3 (TFA salt, 478 mg, 0.6 mmol) and DIPEA (206 μL). The mixture was stirred at room temperature for 10 minutes, and LCMS indicated completion of the reaction. The mixture was then purified directly by reverse-phase HPLC to give compound 1 as a pale yellowish solid (710 mg); 1 H NMR (500 MHz, DMSO-d6) δ 11.41 (s, 1H), 8.25 (d, J = 8.5 Hz, 1H), 8.07 - 7.91 (m, 3H), 7.77 (d, J = 10.9 Hz, 1H), 7.68 - 7.54 (m, 2H), 7.47 (dddd, J = 13.4, 7.6, 4.9, 2.8 Hz, 4H), 7.43 - 7.22 (m, 5H), 6.98 - 6.83 (m, 1H), 5.51 (dt, J = 8.8, 4.5 Hz, 1H), 5.43 (s, 2H), 5.22 (s, 2H), 5.04 (d, J = 14.0 Hz, 1H), 4.47 (q, J = 6.8 Hz, 1H), 4.09 (dp, J = 18.0, 7.1 Hz, 3H), 3.73 (s, 10H), 3.67 - 3.27 (m, 61H), 3.22 - 3.01 (m, 3H), 2.92 (dq, J = 13.3, 6.7 Hz, 1H), 2.68 - 2.45 (m, 8H), 2.45 - 2.28 (m, 6H), 2.19 (dq, J = 9.3, 4.8 Hz, 1H), 1.87 (qd, J = 13.8, 7.1 Hz, 4H), 1.61 (d, J = 13.1 Hz, 2H), 1.49 - 1.21 (m, 7H), 1.13 (dd, J = 26.8, 7.1 Hz, 7H), 0.97 (ddd, J = 16.1, 11.6, 6.4 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H); LCMS m / z (ESI + ): C 93 H 125 FN 10 O 25 calculated for, 1801.9; found 1802.9 [M + H] +

[0859]

[0389] Example 2: Synthesis of DBCO-nnAA-PEG13-VKG-exatecan (Compound 2):

[0860]

[0390]

[0861]

Chem.

[0862]

[0391] To a solution of compound Boc-VK(Fmoc)-G-OH 9 (562 mg, 0.9 mmol) in anhydrous DMF (10 mL) were added exatecan mesylate (1) (478 mg, 0.9 mmol) and DIPEA (470 μL). After all components were dissolved, HATU (342 mg, 0.9 mmol) was added and the mixture was stirred at room temperature for 10 minutes. Then, the reaction was diluted with water (80 mL) and extracted with EtOAc (150 mL). The organic layer was washed with hydrochloric acid (0.2 M, 50 mL) and brine (50 mL), dried over Na2SO4, and evaporated to dryness under reduced pressure. The resulting residue was treated with TFA / DCM (1 / 4, 20 mL) at room temperature for 30 minutes, and the reaction was evaporated to dryness under reduced pressure. The crude mixture was dissolved in 5 mL of DMF and purified by reverse-phase HPLC to give compound 10 as a yellowish solid (771 mg). MS C 52 H 56 Calculated for FN7O9, 941.4; found 942.9 [M+H] +

[0863]

[0392]

[0864]

Chem.

[0865]

[0393] To a solution of compound 4 (426 mg, 0.33 mmol) in anhydrous DMF (3 mL) were added compound 10 (TFA salt, 347 mg, 0.33 mmol) and DIPEA (130 μL). The mixture was stirred at room temperature for 15 minutes, and LCMS indicated the formation of the desired product. Then, DBU (366 μL) was added dropwise and the mixture was stirred at room temperature for an additional 10 minutes. LCMS indicated the completion of the reaction. The mixture was then purified directly by reverse-phase HPLC to give compound 2 as a yellowish solid (350 mg); HRMS m / z (ESI +):C 96 H 133 FN 10 O 24 Calculated value: 1828.94; Measured value: 1829.95 [M+H] + ; 1 1H NMR (500 MHz, DMSO) δ 8.43 (d, J = 8.5 Hz, 1H), 8.14 (t, J = 5.6 Hz, 1H), 7.98 (d, J = 7.4 Hz, 1H), 7.81 (d, J = 10.9 Hz, 1H), 7.77 - 7.53 (m, 7H), 7.54 - 7.23 (m, 10H), 5.57 (dt, J = 8.7, 4.4 Hz, 1H), 5.47 - 5.38 (m, 2H), 5.25 (d, J = 3.3 Hz, 2H), 5.04 (d, J = 14.1 Hz, 2H), 4.16 (td, J = 8.1, 5.9 Hz, 3H), 4.08 (dd, J = 8.5, 6.8 Hz, 3H), 3.88 - 3.68 (m, 13H), 3.50 (d, J = 3.6 Hz, 71H), 3.25 - 3.13 (m, 4H), 3.13 - 3.03 (m, 2H), 2.92 (dq, J = 13.4, 6.8 Hz, 2H), 2.78 (q, J = 6.8 Hz, 3H), 2.60 - 2.26 (m, 18H), 2.26 - 2.03 (m, 3H), 1.98 - 1.74 (m, 6H), 1.74 - 1.46 (m, 8H), 1.46 - 1.09 (m, 12H), 0.97 (s, 2H), 0.88 (t, J = 7.3 Hz, 3H), 0.78 (dd, J = 14.1, 6.7 Hz, 7H).

[0866]

[0394] Example 3: Synthesis of DBCO-nnAA-PEG13-AAA-Exatecan (Compound 3):

[0867]

[0395]

[0868]

Chem.

[0869]

[0396]

[0870]

Chem.

[0871]

[0397] Compound 3 was similarly synthesized using the same method as above. LCMS m / z (ESI + ): C 92 H 124 FN9O 24 Calculated value of, 1757.87; Measured value 1759.1 [M+H] +

[0872]

[0398] Example 4: Synthesis of DBCO-nnAA-PEG13-VK-Exatecan (Compound 4):

[0873]

[0399]

[0874] [Chemical formula]

[0875]

[0400]

[0876] [Chemical formula]

[0877]

[0401] Compound 4 was similarly synthesized using the same method as above. LCMS m / z (ESI + ): C 94 H 130 FN9O 23 Calculated value of, 1771.93; Measured value 1773.1 [M+H] + .

[0878]

[0402] Example 5: Synthesis of DBCO-nnAA-PEG13-NN-Exatecan (Compound 5):

[0879]

[0403]

[0880] [Chemical formula]

[0881]

[0404] A solution of Fmoc-Asn(Trt)-OH 16 (120 mg, 0.2 mmol) and exatecan mesylate 1 (106 mg, 0.2 mmol) in anhydrous DMF (4 mL) was added with PyAOP (110 mg), followed by DIPEA (0.14 mL). When the reaction mixture was stirred at room temperature for 1 hour, LCMS indicated the formation of the desired product. Hydrochloric acid (0.5 M, 30 mL) was added to the reaction mixture. The mixture was extracted with EtOAc (60 mL), the organic layer was dried over Na2SO4, and evaporated to dryness under reduced pressure. The obtained residue was dissolved in DMF (3 mL), and piperidine (0.4 mL) was added. The mixture was stirred at room temperature. After 20 minutes, the mixture was concentrated under reduced pressure to about 2 mL, and the residue was triturated with diethyl ether / hexane (1 / 1, 50 mL) to obtain compound 17. Compound 17 was diluted with DMF (5 mL), and to this mixture was added Fmoc-Asn(Trt)-OH 16 (360 mg, 0.6 mmol), followed by PyAOP (330 mg) and DIPEA (0.21 mL). The mixture was stirred at room temperature for 10 minutes and diluted with EtOAc (100 mL). The mixture was washed with 0.5 M hydrochloric acid (50 mL) and water (50 mL). The organic layer was dried over Na2SO4 and evaporated to dryness under reduced pressure. The obtained gum was treated with TFA / TIS (9 / 1, 5 mL) at room temperature for 20 minutes. 100 mL of diethyl ether was added, and the precipitated solid was collected by filtration to obtain the crude product, which was treated with piperidine (20% in DMF, 3 mL) at room temperature for 10 minutes, and LCMS indicated the completion of the reaction. The mixture was purified by RP-HPLC, and after lyophilization, compound 18 (120 mg) was obtained as a pale yellow solid; LCMS m / z (ESI + ): C 32 H 34 FN7O8 calculated value, 663.25; measured value 664.3 [M+H] +

[0882]

[0405]

[0883]

Chem.

[0884]

[0406] When compound 4 and compound 18 were coupled using the same method as above, compound 5 was obtained as a yellow solid. LCMS m / z(ESI + ):C 91 H 121 FN 10 O 25 Calculated value of, 1772.85; Measured value 1773.91[M+H] +

[0885]

[0407] Example 6. Synthesis of compound 6, compound 7, compound 8, and compound 9:

[0886]

Chemical formula

[0887]

[0408] Compound 6, compound 7, compound 8, and compound 9 were similarly synthesized, purified, and characterized from the common intermediate compound 10 using the same method as above.

[0888]

[0409] Example 7. Synthesis of compound 10:

[0889]

[0410]

[0890]

Chemical formula

[0891]

[0411]

[0892]

Chemical formula

[0893]

[0412]

[0894]

Chemical formula

[0895]

Chem.

[0896]

[0413] General method, resin loading of 2-chlorotrityl chloride resin, Fmoc removal with DMF / piperidine, chain elongation were used to synthesize compound 27 on a 2-chlorotrityl chloride resin solid support. Amide coupling was carried out with HATU, DMF, DIPEA, and the final resin was cleaved by treatment with 2 mL of TFA / DCM (4%). The crude product carboxylic acid was dissolved in DMF (4 mL), Pftu (86 mg, 0.2 mmol) was added, and subsequently DIEA (70 μL) was added. The mixture was stirred at room temperature for 10 minutes and purified directly by reverse-phase HPLC, and compound 27 (185 mg) was obtained as a viscous syrup after lyophilization.

[0897]

[0414]

[0898]

Chem.

[0899]

[0415]

[0900]

Chem.

[0901]

Chem.

[0902]

[0416] To a solution of compound 29 (TFA salt, 40 mg, 44 μmol) and compound 27 (55 mg, 40 μmol) in 2 mL of anhydrous DMF, DIPEA (38 μL) was added and the mixture was stirred at RT for 30 minutes. LCMS indicated the completion of the reaction. The mixture was then purified by reverse-phase HPLC to give compound 10 (52.9 mg) as a yellow solid. LCMS m / z (ESI + ): C 103 H 134 FN9O 31 calculated value for, 2011.92; found 2013.1 [M+H] +

[0903]

[0417] Example 8. Synthesis of compounds 11 and 12:

[0904]

Chem.

[0905]

[0418] Compounds 11 and 12 are synthesized, purified, and characterized in the same manner as compound 10 in the above example using the same method starting from the common intermediate compound 29.

[0906]

[0419] Example 9. Synthesis of compound 13:

[0907]

Chem.

[0908]

[0420] Compound 13 is synthesized in the same manner as compound 2 using the same method as described in Example 2.

[0909]

[0421] Example 10. Synthesis of compound 14:

[0910]

Chem.

[0911]

[0422] Compound 14 is synthesized in the same manner using the same method as compound 1 as described in Example 1.

[0912]

[0423] Example 11. Synthesis of compound 15:

[0913]

Chem.

[0914]

[0424] Compound 15 is synthesized in the same manner using the same method as compound 10 as described in Example 7.

[0915]

[0425] Example 12: Synthesis of compound 16:

[0916]

Chem.

[0917]

[0426] To a solution of compound 28 (44 mg, 51 μmol) in anhydrous DMF (2 mL) were added compound 4 (66 mg, 51 μmol) and DIPEA (18 μL, 156 μmol). The mixture was stirred at room temperature for 20 minutes, and LCMS indicated completion of the reaction. The mixture was then purified directly by reverse-phase HPLC to give compound 16 as a yellow solid (61 mg). LCMS m / z (ESI + ): Calculated for C 98 H 127 FN 10 O 25 1862.90; Found 1864.1 [M+H] +

[0918]

[0427] Example 13: Synthesis of compound 17:

[0919]

Chem.

[0920]

[0428] Compound 17 is synthesized in the same manner using the same method as Compound 16 as described in Example 12.

[0921]

[0429] Example 14: Synthesis of Compound 18

[0922]

Chem.

[0923]

[0430] Compound 18 is synthesized in the same manner using the same method as Compound 16 as described in Example 12.

[0924]

[0431] Example 15. Synthesis of Compounds 19, 20, 21, 22, and 23:

[0925]

Chem.

[0926]

Chem.

[0927]

[0432] Compounds 19, 20, 21, 22, and 23 were synthesized using the same method as above.

[0928]

[0433] Example 16. Conjugation of Linker-Payload Compounds

[0929]

[0434] The low-molecular-weight drug linker was dissolved in DMSO to a final concentration of 5 mM. Conjugation was carried out in 1×PBS at an antibody concentration of 1 mg / mL, a drug linker:pAMF ratio of 3, and 15% DMSO. The reaction mixture was incubated overnight at 30 °C. The conjugation efficiency was measured by MALDI. The unconjugated drug linker was removed by desalting. The purity of the conjugate was measured by Sepax SEC-300. The conjugate was formulated in 1×PBS.

[0930]

[0435] The linker drug was conjugated to the aFolR mAb by incorporating four pAMF sites and eight pAMF sites into the heavy-chain Y180F404 site and the light-chain K42E161 site. According to the above conjugation conditions, a conjugation efficiency of over 94% was achieved for all linker-payloads. The results of analytical SEC showed that all conjugates were over 99% monomer and showed high purity (Table 1).

[0931]

Table 3

[0932]

[0436] Example 17: Cell-killing activity of free drug

[0933]

[0437] The in vitro cell-killing activities of exatecan, gly-exatecan, and hemiasterlin were evaluated in a panel of breast cancer cell lines and lung cancer cell lines.

[0934]

Chemical formula

[0935]

[0438] MDA-MB-361, SKBR3, BT20, HCC38, HCC1143, HCC1937, JIMT1, MCF-7, A549, NCI-H441, NCI-H520, NCI-H1703, NCI-H1975, NCI-H2110, NCI-H226, NCI-H292, and NCI-H358 cells were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA). All cell lines were maintained in DMEM:F12 (1:1), high glucose (Corning, Corning, NY) supplemented with 10% heat-inactivated fetal bovine serum (Thermo Scientific, Grand Island, NY), 2 mM glutamax (Thermo Scientific, Grand Island, NY), and 1× penicillin / streptomycin (Corning, Corning, NY). The cytotoxic effect of the exatecan warhead was measured in a cell proliferation assay. Cells were seeded at 625 cells / 25 μL into 384-well flat-bottom white polystyrene plates the day before the actual assay began. Exatecan was formulated in cell culture medium at 2× starting concentration and serially diluted (1:3) under sterile conditions and added to the cells in triplicate. Plates were incubated at 37 °C for 120 h in a CO2 incubator. For cell viability measurements, 30 μL of Cell Titer-Glo® reagent (Promega Corp, Madison, WI) was added to each well and the plates were processed according to the product instructions. Relative luminescence was measured with an ENVISION® plate reader (Perkin-Elmer; Waltham, MA). Relative luminescence readings were converted to % viability using untreated cells as a control. Data were fitted with a four-parameter fitting equation using nonlinear regression analysis with log (inhibitor) vs. response, variable slope, GraphPad Prism.

[0936]

[0439] The exatecan-free warhead, like hemiasterlin, had an EC in the range of 0.32 nM to 4.2 nM 50Values (Table 2) showed potent cell killing against all cell lines tested. Gly-exatecan had an EC in the range of 3.8 nM to 44 nM 50 Values (Table 2) showed slightly lower cell killing activity.

[0937]

Table 4

[0938]

[0440] Example 18: In Vitro Cell Killing Activity of Exatecan ADC

[0939]

[0441] Anti-FolRa ADCs were generated by conjugating anti-FolRa antibody 1848-H01 with exatecan linker warheads at DAR = 4 and DAR = 8. Anti-FolRa antibody 1848-H01 conjugated with compound 27 and compound 26 at DAR = 4 and DAR = 8 was used as a positive control.

[0940]

Table 5

[0941]

[0442] FolRα-positive Igrov1 cells were licensed from the NCI (National Cancer Institute at Frederick, Maryland). FolRα-negative A549 cells were purchased from the ATCC (American Type Culture Collection). Both cell lines were maintained in DMEM:F12 (1:1), high glucose (Corning) supplemented with 10% heat-inactivated fetal bovine serum (Thermo Scientific), 2 mM glutamax (Thermo Scientific), and 1× penicillin / streptomycin (Corning). The cytotoxic effect of the ADC was measured by a cell proliferation assay. Igrov1 cells and A549 cells at a concentration of 625 cells / 25 μl were seeded onto 384-well flat-bottom white polystyrene plates the day before the assay. The ADC was formulated in cell culture medium at 2× the starting concentration and sterile filtered through a SpinX 0.22 μm filter centrifugal tube (Corning Costar). The filter-sterilized sample was serially diluted (1:3) under sterile conditions and added to the cells in triplicate. The plates were incubated at 37 °C in a CO2 incubator for 120 hours. For measurement of cell viability, 30 microliters of Cell Titer-Glo® reagent (Promega Corp, Madison, Wisconsin) was added to each well and the plates were processed according to the product instructions. Relative luminescence was measured with an ENVISION® plate reader (Perkin-Elmer; Waltham, Massachusetts). The relative luminescence readings were converted to % viability using untreated cells as a control. The data were fitted with a four-parameter fitting equation using non-linear regression analysis, logarithmic (inhibitor) vs. response, variable slope, GraphPad Prism.

[0942]

[0443] As shown in FIGS. 1A and 1B, anti-FolRα ADCs conjugated with compound 1, compound 2, and compound 3 at DAR = 4 and DAR 8 showed potent cell killing against FolRα-positive Igrov1 cells, but no cell killing was observed in FolRα-negative A549 cells. This indicates that there was no non-specific release of free warheads that killed target-negative cells, meaning that all the linkers tested were stable in cell culture medium for 5 days. Compared with the ADCs with DAR4, the ADCs with DAR8 showed more potent cell killing with lower EC 50 and a larger killing range.

[0943]

[0444] As shown in FIGS. 2A and 2B, anti-FolRa ADCs conjugated with compound 6, compound 7, compound 8, and compound 9 at DAR = 8 showed potent cell killing against FolRα-positive Igrov1 cells, but no cell killing was observed in hFolRα-negative A549 cells. The anti-FolRα ADC conjugated with compound 24 showed no cell killing against either FolRα-positive Igrov1 cells or FolRα-negative A549 cells.

[0944]

[0445] The cell killing activities of exatecan ADCs are summarized in Tables 3 and 4.

[0945]

Table 6

[0946]

Table 7

[0947]

[0446] Example 19: In Vitro Cell Killing Activity of Hemiasparin ADC

[0948]

[0447] Anti-FolRα ADCs conjugated with hemiasterlin using different cathepsin-cleavable linkers were also generated at DAR = 4, and their cell killing activities were evaluated in FolRα-positive Igrov1 cells and FolRα-negative A549 cells. As shown in FIGS. 3A, 3B, and Table 5, similar to anti-FolRα ADC conjugate 127-3, the anti-FolRα ADCs conjugated with compound 13, compound 14, and compound 15 at DAR 4 showed potent cell killing against FolRα-positive Igrov1 cells, but no cell killing was observed in FolRα-negative A549 cells, indicating that all the linkers tested were stable in cell culture medium for 5 days.

[0949]

Table 8

[0950]

[0448] The disclosure set forth above may embrace a plurality of distinct embodiments having independent utility. Although each of these embodiments is disclosed, since numerous modifications are possible, the specific embodiments disclosed and illustrated herein should not be considered in a limiting sense. The subject matter of the embodiments includes all novel and non-obvious combinations and sub-combinations of the various elements, features, functions, and / or characteristics disclosed herein. The following claims particularly point out certain combinations and sub-combinations that are regarded as novel and non-obvious. Alternative embodiments such as other combinations and sub-combinations of features, functions, elements, and / or characteristics may be claimed in this application, an application claiming priority from this application, or a related application. Such claims, whether for different embodiments or the same embodiment, and whether broader, narrower, equal, or different in scope compared to the original claims, are considered to be included within the subject matter of this disclosure.

[0951] One or more features of any embodiment described in this specification or the drawings may be combined with one or more features of any other embodiment described in this specification or the drawings without departing from the scope of the present disclosure.

[0952]

[0450] All publications, patents, and patent applications cited herein are hereby incorporated by reference as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. The foregoing disclosure has been described in some detail by way of example and for purposes of clarity of understanding, but it will be readily apparent to those of ordinary skill in the art that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

Claims

1. A compound of formula (I): 【Chemical 1】 or a pharmaceutically acceptable salt and / or stereoisomer thereof wherein L １ is [Chemical 2] is selected from; Ring A is an optionally substituted bridged, fused, or spirocyclic bicyclic carbocycle or an optionally substituted bridged, fused, or spirocyclic bicyclic heterocycle, and the carbocycle or heterocycle of Ring A is selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)-, -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , optionally substituted with one or more substituents selected from aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; Ring B is an optionally substituted N-linked bridged, fused, or spirocyclic bicyclic heterocycle, where Ring B is an alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)-, -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , optionally substituted with one or more substituents selected from aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; R ａ and R ｂ is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; a is an integer independently selected from 0, 1, 2, 3, 4, 5, and 6; b is an integer selected from 0 and 1; R １ is alkyl optionally substituted with one or more substituents selected from hydrogen, or cycloalkyl, halogen, alkoxy, -CN, -NO ２ , -OH, -N(R ２ R ３ ), -C(O)N(R ２ R ２ R ３ ), -C(O)OR ２ , aryl, and heteroaryl; ２ and is optionally substituted with one or more substituents selected from hydrogen, or cycloalkyl, halogen, alkoxy, -CN, -NO R ２ and R ３ is independently selected from hydrogen, alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; Y is *-C(O)-(CR ａ R ｂ ) ｃ -NH- or *-C(O)-(CR ａ R ｂ ) ｃ -, where * represents the location where Y is attached to RG; c is an integer selected from 1, 2, 3, 4, 5, and 6; RG is a reactive group; L ２ is non-existent or is a linker containing a hydrophilic polymer residue; L ３ is absent, -C(O)-AA-, -C(O)-AA-Z-(CR ａ R ｂ ) ａ -Z-(CR ａ R ｂ ) ａ -C(O)-, -C(O)-Z-(CR ａ R ｂ ) ａ -C(O)-Z-L ４ -OC(O)-, -Z-AA-, -AA-, -C(O)-, -C(O)-AA-Z-(CR ａ R ｂ ) ａ -, -AA-C(O)-, -C(O)-(CR ａ R ｂ ) ａ -Z-(CR ａ R ｂ ) ａ -Z-AA-C(O)-, -C(O)O-L ４ -Z-C(O)-(CR ａ R ｂ ) ａ -Z-C(O)-, -AA-Z-, or -(CR ａ R ｂ ) ａ -Z-AA-C(O)-; Z is selected from -NR ２ - and -O-; AA is an amino acid residue or a peptide residue; L ４ is 【Chemical Formula 3】 and Su is a hexose form of a monosaccharide; d is an integer independently selected from 1, 2, and 3; D is a cytotoxic payload; 【Chemical Formula 4】 represents a bond to the remainder of the compound).

2. L １ is 【Chemical Formula 5】 The compound according to claim 1, wherein

3. L １ is [Chemical Formula 6] The compound according to claim 2, wherein

4. L １ is 【Chemical Formula 7】 The compound according to claim 1, wherein

5. L １ wherein ring A is an optionally substituted bridged, fused, or spirocyclic bicyclic carbocycle, and the carbocycle of ring A is １～１２ Alkyl, C ２～１２ Alkenyl, C ２～１２ Alkynyl, C ３～１２ Cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)-, -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ 5. The compound according to claim 1 , optionally substituted with one or more substituents selected from: aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

6. L １ wherein the ring A of L is an optionally substituted C ４～１２ bridged, fused, or spiro bicyclic carbocyclic ring, the compound according to claim 5.

7. L １ wherein the ring A of １ is an optionally substituted C ４～１２ bridged bicyclic carbocyclic ring, the compound according to claim 5.

8. L １ wherein the ring A of L is an optionally substituted C ４～８ bridged bicyclic carbocyclic ring; the compound according to claim 5.

9. L １ is 【Chemical Formula 8】 The compound according to claim 1 or 4, wherein

10. L １ is 【Chemical Formula 9】 The compound according to claim 9, wherein

11. L １ is 【Chemical Formula 10】 The compound according to claim 9, wherein

12. L １ is 【Chemical 11】 The compound according to claim 1, wherein

13. L １ wherein ring B is an optionally substituted 5-12 membered N-linked bridged, fused, or spirocyclic bicyclic heterocycle containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, including the N to which the ring is attached, and the heterocycle of ring B is １～１２ Alkyl, C ２～１２ Alkenyl, C ２～１２ Alkynyl, C ３～１２ Cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)-, -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ 13. The compound of claim 1 or 12, optionally substituted with one or more substituents selected from: aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

14. L １ The compound according to claim 13, wherein the ring B of L is an optionally substituted 5- to 12-membered N-bonded spirobicyclic heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, including N to which the ring is bonded.

15. L １ is 【Chemical 12】 is selected from, and m is an integer selected from 1, 2, 3, 4, and 5; each of n and o is an integer independently selected from 1, 2, and 3, the compound according to claim 1 or 12.

16. L １ is 【Chemical 13】 is selected from, and m is an integer selected from 1, 2, 3, 4, and 5; each of n and o is an integer independently selected from 1, 2, and 3, the compound according to claim 1 or 12.

17. L １ The ring B of 【Chemical 14】 selected from, X １ X ２ X ３ and X ４ are independently selected from -C(R ４ ) ２ -, -NH-, -O-, and -S-, and when X １ X ２ and X ３ are present, at least one of X １ to X ３ is -C(R ４ ) ２ -, and when X １ X ２ X ３ and X ４ are present, at least two of X １ to X ４ are -C(R ４ ) ２ -; R ４ is hydrogen, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～１２ Cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; or two R on the same carbon are independently selected from ４ 13. The compound of claim 1 or 12, wherein the groups taken together form an oxo group.

18. Ring B is 【Chemical Formula 15】 selected from, the compound according to claim 17.

19. Ring B is 【Chemical 16】 selected from, X １ X ２ X ３ and X ４ are independently selected from -C(R ４ ) ２ -, -NH-, -O-, and -S-, and when X １ X ２ and X ３ are present, at least one of X １ to X ３ is -C(R ４ ) ２ -, and when X １ X ２ X ３ and X ４ are present, at least two of X １ to X ４ are -C(R ４ ) ２ -; R ４ is independently selected from hydrogen, C １～６ alkyl, C ２～６ alkenyl, C ２～６ alkynyl, C ３～１２ cycloalkyl, halogen, alkoxy, -CN, -NO ２ , -OH, -N(R ２ R ３ ), -C(O)N(R ２ R ２ R ３ ), -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; or two R ２ groups on the same carbon together form an oxo group, the compound according to claim 1 or 12. ４ ​

20. Ring B is 【Chemical 17】 selected from, the compound according to claim 19.

21. Ring A is 【Chemical Formula 18】 selected from, X １ X ２ X ３ and X ４ are independently selected from -C(R ４ ) ２ -, -NH-, -O-, and -S-, and when X １ X ２ and X ３ are present, at least one of X １ to X ３ is -C(R ４ ) ２ -, and when X １ X ２ X ３ and X ４ are present, at least two of X １ to X ４ are -C(R ４ ) ２ -; X ５ is -CR ４ - or -N-; R ４ is independently selected from hydrogen, C １～６ alkyl, C ２～６ alkenyl, C ２～６ alkynyl, C ３～１２ cycloalkyl, halogen, alkoxy, -CN, -NO ２ , -OH, -N(R ２ R ３ ), -C(O)N(R ２ R ２ R ３ ), -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; or two R ２ groups on the same carbon together form an oxo group, the compound according to any one of claims 1 to 4. ４ ​

22. A compound of formula (III): 【Chemical Formula 19】 or a pharmaceutically acceptable salt and / or stereoisomer thereof wherein L ５ is a linker containing a non-natural amino acid; Y is *-C(O)-(CR ａ R ｂ ) ｃ -NH- or *-C(O)-(CR ａ R ｂ ) ｃ -, where * represents the location where Y is attached to RG; c is an integer selected from 1, 2, 3, 4, 5, and 6; RG is a reactive group; L ２ is non-existent or is a linker containing a hydrophilic polymer residue; L ３ is absent, -C(O)-AA-, -C(O)-AA-Z-(CR ａ R ｂ ), ａ -Z-(CR ａ R ｂ ), ａ -C(O)-, -C(O)-Z-(CR ａ R ｂ ), ａ -C(O)-Z-L ４ -OC(O)-, -Z-AA-, -AA-, -C(O)-, -C(O)-AA-Z-(CR ａ R ｂ ), ａ -, -AA-C(O)-, -C(O)-(CR ａ R ｂ ), ａ -Z-(CR ａ R ｂ ), ａ -Z-AA-C(O)-, -C(O)O-L ４ -Z-C(O)-(CR ａ R ｂ ), ａ -Z-C(O)-, -AA-Z-, or -(CR ａ R ｂ ), ａ -Z-AA-C(O)-; R ａ and R ｂ are each independently selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO ２ , -OH, -N(R ２ R ３ ), -C(O)N(R ２ R ２ R ３ ), -C(O)OR ２ , ２ aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl;​ a is an integer independently selected from 0, 1, 2, 3, 4, 5, and 6; Z is selected from -NR ２ - and -O-; AA is an amino acid residue or a peptide residue; L ４ is 【Chemical 20】 and Su is a hexose form of a monosaccharide; d is an integer independently selected from 1, 2, and 3; D is a cytotoxic payload; 【Chemical 21】 represents a bond to the remainder of the compound).

23. L ５ The compound according to claim 22, wherein L is a linker containing at least one amino acid selected from sulfonalanine, hydroxyproline (Hyp), β-alanine, citrulline (Cit), ornithine (Orn), norleucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, and methionine sulfone.

24. L ５ is 【Chemical 22】 The compound according to claim 22, wherein the linker comprises

25. L ５ is 【Chemical 23】 The compound according to claim 22, which is

26. L ２ is -(CR ａ R ｂ ) ａ -POLY １ -, -POLY １ -, -(CR ａ R ｂ ) ａ -POLY １ -(CR ａ R ｂ ) ａ -, 【Chemical 24】 selected from, POLY １ is a divalent hydrophilic polymer residue; POLY ２ is a residue of an aqueous polymer; 【Chemical Formula 25】 The compound according to any one of claims 1 to 25, wherein represents a bond to the remainder of the compound.

27. Y is -C(O)-(CR ａ R ｂ ) ｃ -NH-, the compound according to any one of claims 1 to 26.

28. Y is -C(O)-CH ２ CH ２ -NH- and the compound according to claim 27.

29. Y is -C(O)-(CR ａ R ｂ ) ｃ - and is a compound according to any one of claims 1 to 26.

30. Y is -C(O)-(CH ２ ) ４ - and is the compound according to claim 29.

31. The compound according to any one of claims 1 to 30, wherein RG comprises an alkyne, cyclooctyne, strained alkene, tetrazine, methylcyclopropene (Mecyp), thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide.

32. RG is 【Chemical 26】 -N ３ 、-NH ２ 、and -SH; R Ｔ is C １～６ alkyl; 【Chemical 27】 The compound according to claim 31, wherein represents a bond to the remainder of the compound.

33. RG is 【Chemical Formula 28】 The compound according to claim 32, which is selected from

34. RG is ​ The compound according to claim 32, which is

35. L ２ is -(CR ａ R ｂ ) ａ -POLY １ - and is a compound according to any one of claims 1 to 25 and 31 to 34.

36. -POLY １ - is a divalent residue of polyethylene glycol (PEG), poly(propylene glycol) (PPG), or a copolymer of ethylene glycol and propylene glycol, the compound according to claim 35.

37. The divalent hydrophilic polymer residue has the formula 【Chemical Formula 30】 having, R ５ The compound according to claim 36, wherein R is hydrogen or methyl and x is an integer between 1 and 100 inclusive at both ends.

38. 【Fig. 31】 is 【Chemical 32】 The compound according to claim 37, which is

39. The compound according to claim 36 or 37, wherein x is an integer between 1 and 25 inclusive at both ends.

40. The compound according to claim 36 or 37, wherein x is an integer between 1 and 15 inclusive at both ends.

41. The compound according to claim 36 or 37, wherein x is 13.

42. L ２ is 【Chemical 33】 The compound according to any one of claims 1 to 25, which is

43. POLY ２ The compound according to claim 42, wherein POLY is a residue of polyethylene glycol (PEG), methoxypolyethylene glycol (mPEG), poly(propylene glycol) (PPG), a copolymer of ethylene glycol and propylene glycol, or polysarcosine.

44. POLY ２ is 【Chemical Formula 34】 and R ５ The compound according to claim 43, wherein R is hydrogen or methyl and x is an integer between 1 and 100 inclusive at both ends.

45. 【Fig. 35】 is 【Chemical 36】 The compound according to claim 44, which is

46. The compound according to claim 43 or 44, wherein x is an integer between 1 and 25 inclusive at both ends.

47. The compound according to claim 43 or 44, wherein x is an integer between 1 and 15 inclusive at both ends.

48. The compound according to claim 43 or 44, wherein x is 11.

49. L ３ is absent, -C(O)-AA-, -C(O)-AA-Z-(CR ａ R ｂ ), ａ -Z-(CR ａ R ｂ ), ａ -C(O)-, -C(O)-Z-(CR ａ R ｂ ), ａ -C(O)-Z-L ４ -OC(O)-, -Z-AA-, -AA-, or -C(O)-, a compound according to any one of claims 1 to 48.

50. L ３ The compound according to any one of claims 1 to 48, wherein L is -C(O)-AA-.

51. L ３ is -C(O)-AA-Z-(CR ａ R ｂ ), ａ -Z-(CR ａ R ｂ ), ａ -C(O)-, -Z-AA-, or -AA-, a compound according to any one of claims 1 to 48.

52. The compound according to claim 51, wherein Z is -NH-.

53. L ３ The compound according to any one of claims 1 to 48, wherein L is -AA-.

54. -AA- is 【Chemical 37】 The compound according to claim 53, which is

55. The compound according to any one of claims 50 to 53, wherein AA is a dipeptide residue, tripeptide residue, tetrapeptide residue, or pentapeptide residue.

56. The compound according to claim 55, wherein AA is a tripeptide residue.

57. The compound according to any one of claims 50 to 53 and 55 to 56, wherein AA comprises at least one amino acid residue selected from alanine, glycine, valine, and asparagine.

58. The compound according to any one of claims 50 to 53 and 55 to 56, wherein AA comprises at least one amino acid residue selected from alanine and glycine.

59. wherein AA is 【Chemical Formula 38】 The compound according to any one of claims 50 to 53, selected from the group consisting of

60. L ３ is -C(O)-Z-(CR ａ R ｂ ) ａ -C(O)-Z-L ４ -OC(O)-; L ４ is 【Chemical Formula 39】 and; The compound according to any one of claims 1 to 48, wherein Su is a hexose type of monosaccharide.

61. wherein Su is 【Chemical Formula 40】 The compound according to claim 60.

62. wherein Su is 【Chemical Formula 41】 The compound according to claim 61.

63. The compound according to any one of claims 1 to 62, wherein D is a cytotoxic payload selected from the group consisting of a tubulin inhibitor, a DNA damaging agent, a DNA topoisomerase I inhibitor, a DNA topoisomerase II inhibitor, and an RNA polymerase II inhibitor.

64. The compound according to claim 63, wherein D is a cytotoxic payload selected from the group consisting of hemiasterlin or a derivative thereof, camptothecin or a derivative thereof, anthracycline or a derivative thereof, PNU-159682 or a derivative thereof, PBD or a derivative thereof, and duocarmycin or a derivative thereof.

65. wherein D is 【Chemical 42】 The compound according to claim 63, selected from or derivatives thereof.

66. The compound according to claim 63, wherein D is an exatecan payload.

67. 【Chemical Formula 43】 The compound according to claim 1, represented by or a pharmaceutically acceptable salt thereof.

68. 【Chemical Formula 44】 The compound according to claim 67, represented by or a pharmaceutically acceptable salt thereof.

69. 【Chemical Formula 45】 The compound according to claim 67, represented by or a pharmaceutically acceptable salt thereof.

70. 【Fig. 46】 The compound according to claim 1, represented by or a pharmaceutically acceptable salt thereof.

71. 【Fig. 47】 The compound according to claim 70, represented by or a pharmaceutically acceptable salt thereof.

72. 【Chemical Formula 48】 The compound according to claim 71, represented by or a pharmaceutically acceptable salt thereof.

73. 【Fig. 49-1】 【Chemical 49-2】 【Chemical Formula 49-3】 【Chemical Formula 49-4】 【Chemical Formula 49-5】 【Chemical Formula 49-6】 【Chemical Formula 49-7】 【Chemical Formula 49-8】 The compound according to claim 1, selected from or a pharmaceutically acceptable salt thereof.

74. 【Fig. 50-1】 【Chemical Formula 50-2】 【Chemical Formula 50-3】 【Chemical Formula 50-4】 【Chemical Formula 50-5】 【Chemical Formula 50-6】 【Chemical Formula 50-7】 【Chemical Formula 50-8】 The compound according to claim 73, selected from or a pharmaceutically acceptable salt thereof.

75. 【Fig. 51】 The compound according to claim 72, selected from or a pharmaceutically acceptable salt thereof.

76. Formula: 【Chemical 52】 The compound according to claim 22, or a pharmaceutically acceptable salt thereof.

77. Formula: 【Chemical 53】 The compound according to claim 76, or a pharmaceutically acceptable salt thereof.

78. A conjugate comprising a compound according to any one of claims 1 to 77, or a pharmaceutically acceptable salt thereof, linked to a second compound.

79. Formula II: 【Chemical 54】 or a pharmaceutically acceptable salt thereof (wherein, L １ is 【Chemical Formula 55】 selected from; Ring A is an optionally substituted bridged, fused, or spirocyclic bicyclic carbocycle or an optionally substituted bridged, fused, or spirocyclic bicyclic heterocycle, and the carbocycle or heterocycle of Ring A is selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)-, -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , optionally substituted with one or more substituents selected from aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; Ring B is an optionally substituted N-linked bridged, fused, or spirocyclic bicyclic heterocycle, where Ring B is an alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)-, -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , optionally substituted with one or more substituents selected from aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; R ａ and R ｂ is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; a is an integer independently selected from 0, 1, 2, 3, 4, 5, and 6; b is an integer selected from 0 and 1; R １ is alkyl optionally substituted with one or more substituents selected from hydrogen, or cycloalkyl, halogen, alkoxy, -CN, -NO ２ , -OH, -N(R ２ R ３ ), -C(O)N(R ２ R ２ R ３ ), -C(O)OR ２ , aryl, and heteroaryl; ２ and is optionally substituted with one or more substituents selected from hydrogen, or cycloalkyl, halogen, alkoxy, -CN, -NO, -OH, -N(R)R, -C(O)N(R)R, -C(O)OR, aryl, and heteroaryl; R ２ and R ３ is independently selected from hydrogen, alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; Y is *-C(O)-(CR ａ R ｂ ) ｃ -NH- or *-C(O)-(CR ａ R ｂ ) ｃ -, where * represents the location where Y is attached to RL; c is an integer selected from 1, 2, 3, 4, 5, and 6; RL is a reactive residue; L ２ is non-existent or is a linker containing a hydrophilic polymer residue; L ３ is absent, -C(O)-AA-, -C(O)-AA-Z-(CR ａ R ｂ ) ａ -Z-(CR ａ R ｂ ) ａ -C(O)-, -C(O)-Z-(CR ａ R ｂ ) ａ -C(O)-Z-L ４ -OC(O)-, -Z-AA-, -AA-, -C(O)-, -C(O)-AA-Z-(CR ａ R ｂ ) ａ -, -AA-C(O)-, -C(O)-(CR ａ R ｂ ) ａ -Z-(CR ａ R ｂ ) ａ -Z-AA-C(O)-, -C(O)O-L ４ -Z-C(O)-(CR ａ R ｂ ) ａ -Z-C(O)-, -AA-Z-, or -(CR ａ R ｂ ) ａ -Z-AA-C(O)-; Z is selected from -NR ２ - and -O-; AA is an amino acid residue or a peptide residue; L ４ is 【Chemical 56】 and Su is a hexose type of monosaccharide; d is an integer independently selected from 1, 2, and 3; D is a cytotoxic payload; COMP is a residue of a second compound; 【Chemical 57】 represents a bond to the remainder of the compound) The conjugate according to claim 78.

80. Formula (IV): 【Chemical Formula 58】 or a pharmaceutically acceptable salt and / or positional isomer thereof (wherein, L ５ is a linker containing a non-natural amino acid; Y is *-C(O)-(CR ａ R ｂ ) ｃ -NH- or *-C(O)-(CR ａ R ｂ ) ｃ -, where * represents the location where Y is attached to RL; c is an integer selected from 1, 2, 3, 4, 5, and 6; RL is a reactive residue; L ２ is non-existent or is a linker containing a hydrophilic polymer residue; L ３ is absent, -C(O)-AA-, -C(O)-AA-Z-(CR ａ R ｂ ) ａ -Z-(CR ａ R ｂ ) ａ -C(O)-, -C(O)-Z-(CR ａ R ｂ ) ａ -C(O)-Z-L ４ -OC(O)-, -Z-AA-, -AA-, -C(O)-, -C(O)-AA-Z-(CR ａ R ｂ ) ａ -, -AA-C(O)-, -C(O)-(CR ａ R ｂ ) ａ -Z-(CR ａ R ｂ ) ａ -Z-AA-C(O)-, -C(O)O-L ４ -Z-C(O)-(CR ａ R ｂ ) ａ -Z-C(O)-, -AA-Z-, or -(CR ａ R ｂ ) ａ -Z-AA-C(O)-; R ａ and R ｂ is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; a is an integer independently selected from 0, 1, 2, 3, 4, 5, and 6; Z is selected from -NR ２ - and -O-; AA is an amino acid residue or a peptide residue; L ４ is 【Chemical Formula 59】 and Su is a hexose type of monosaccharide; d is an integer independently selected from 1, 2, and 3; D is a cytotoxic payload; COMP is a residue of a second compound; 【Chemical 60】 represents a bond to the remainder of the compound) The conjugate according to claim 78.

81. The conjugate according to claim 79 or 90, wherein COMP is a residue of a polypeptide.

82. The conjugate according to claim 79 or 80, wherein COMP is a residue of an antibody.

83. The conjugate according to claim 79 or 80, wherein COMP is a residue of an antibody chain.

84. L １ is 【Chemical Formula 61】 The conjugate according to any one of claims 79 and 81 to 83, wherein

85. L １ wherein ring A is an optionally substituted bridged, fused, or spirocyclic bicyclic carbocycle, and the carbocycle of ring A is １～１２ Alkyl, C ２～１２ Alkenyl, C ２～１２ Alkynyl, C ３～１２ Cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)-, -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ 84. The conjugate of any one of claims 79 and 81 to 83, optionally substituted with one or more substituents selected from aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

86. L １ wherein the ring A of L is an optionally substituted C ４～１２ bridged, fused, or spiro bicyclic carbocyclic ring, the conjugate according to claim 85.

87. L １ wherein the ring A of １ is an optionally substituted C ４～１２ bridged bicyclic carbocyclic ring, the conjugate according to claim 86.

88. L １ wherein the ring A of １ is an optionally substituted C ４～８ bridged bicyclic carbocyclic ring, the conjugate according to claim 87.

89. L １ is 【Chemical Formula 62】 The conjugate according to any one of claims 79 and 81 to 83, wherein

90. L １ wherein ring B is an optionally substituted 5-12 membered N-linked bridged, fused, or spirocyclic bicyclic heterocycle containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, including the N to which the ring is attached, and the heterocycle of ring B is １～１２ Alkyl, C ２～１２ Alkenyl, C ２～１２ Alkynyl, C ３～１２ Cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)-, -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ 84. The conjugate of any one of claims 79 and 81 to 83, optionally substituted with one or more substituents selected from aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl.

91. L １ The ring B of １ is an optionally substituted 5- to 12-membered N-bonded spirobicyclic heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from N, O, and S, including N to which the ring is bonded. The conjugate according to any one of claims 79 and 81 to 83.

92. L １ is 【Chemical Formula 63】 selected from, and m is an integer selected from 1, 2, 3, 4, and 5; The conjugate according to any one of claims 79 and 81 to 83, wherein each of n and o is an integer independently selected from 1, 2, and 3.

93. L １ is 【Chemical Formula 64】 selected from, and m is an integer selected from 1, 2, 3, 4, and 5; The conjugate according to any one of claims 79 and 81 to 83, wherein each of n and o is an integer independently selected from 1, 2, and 3.

94. Ring B is 【Chemical Formula 65】 selected from, X １ X ２ X ３ and X ４ are independently selected from -C(R ４ ) ２ -, -NH-, -O-, and -S-, and when X １ X ２ and X ３ are present, at least one of X １ to X ３ is -C(R ４ ) ２ -, and when X １ X ２ X ３ and X ４ are present, at least two of X １ to X ４ are -C(R ４ ) ２ -; R ４ is independently selected from hydrogen, C １～６ alkyl, C ２～６ alkenyl, C ２～６ alkynyl, C ３～１２ cycloalkyl, halogen, alkoxy, -CN, -NO ２ , -OH, -N(R ２ R ３ ), -C(O)N(R ２ R ２ R ３ ), -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; or two R ２ groups on the same carbon together form an oxo group; a conjugate according to any one of claims 79 and 81 to 83. ４ ​

95. Ring B is 【Chemical Formula 66】 selected from, the conjugate according to claim 94.

96. Ring B is 【Chemical Formula 67】 selected from, X １ X ２ X ３ and X ４ are independently selected from -C(R ４ ) ２ -, -NH-, -O-, and -S-, and when X １ X ２ and X ３ are present, at least one of X １ to X ３ is -C(R ４ ) ２ -, and when X １ X ２ X ３ and X ４ are present, at least two of X １ to X ４ are -C(R ４ ) ２ -; R ４ is hydrogen, C １～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～１２ Cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; or two R on the same carbon are independently selected from ４ 84. The conjugate of any one of claims 78 and 81-83, wherein the groups together form an oxo group.

97. Ring B is 【Chemical Formula 68】 selected from, the conjugate according to claim 96.

98. Ring A is 【Chemical Formula 69】 selected from, X １ X ２ X ３ and X ４ are independently selected from -C(R ４ ) ２ -, -NH-, -O-, and -S-, and when X １ X ２ and X ３ are present, at least one of X １ to X ３ is -C(R ４ ) ２ -, and when X １ X ２ X ３ and X ４ are present, at least two of X １ to X ４ are -C(R ４ ) ２ -; X ５ is N or CR ４ ; R ４ is hydrogen, C ２～６ Alkyl, C ２～６ Alkenyl, C ２～６ Alkynyl, C ３～１２ Cycloalkyl, halogen, alkoxy, -CN, -NO ２ , —OH, —N(R ２ R ３ ) ２ , -C(O)N(R ２ R ３ ) ２ , -C(O)OR ２ , aminoalkyl, hydroxyalkyl, haloalkyl, aryl, arylalkyl, heteroaryl, and heteroarylalkyl; or two R on the same carbon are independently selected from ４ 84. The conjugate of any one of claims 79 and 81-83, wherein the groups together form an oxo group.

99. L １ is 【Chemical Formula 70】 is, the conjugate according to any one of claims 79 and 81 to 83.

100. L １ is 【Chemical Formula 71】 is, the conjugate according to claim 99.

101. L ５ The conjugate according to claim 80, wherein L is a linker containing at least one amino acid selected from sulfonalanine, hydroxyproline (Hyp), β-alanine, citrulline (Cit), ornithine (Orn), norleucine (Nle), 3-nitrotyrosine, nitroarginine, pyroglutamic acid (Pyr), naphthylalanine (Nal), 2,4-diaminobutyric acid (DAB), methionine sulfoxide, and methionine sulfone.

102. L ５ is 【Chemical 72】 is a linker containing, the conjugate according to claim 101.

103. L ５ is 【Chemical 73】 is, the conjugate according to claim 102.

104. L ２ is -(CR ａ R ｂ ) ａ -POLY １ -, -POLY １ -, -(CR ａ R ｂ ) ａ -POLY １ -(CR ａ R ｂ ) ａ -, 【Chemical Formula 74】 selected from, POLY １ is a divalent hydrophilic polymer residue, POLY ２ is a residue of a hydrophilic polymer; 【Chemical Formula 75】 represents a bond to the remainder of the compound, the conjugate according to any one of claims 79 to 103.

105. Y is -C(O)-(CR ａ R ｂ ) ｃ -NH- and the conjugate according to any one of claims 79 to 104.

106. Y is -C(O)-CH ２ CH ２ -NH-, the conjugate according to claim 105.

107. Y is -C(O)-(CR ａ R ｂ ) ｃ - and is the conjugate according to any one of claims 79 to 104.

108. Y is -C(O)-(CH ２ ) ４ - and is the conjugate according to claim 107.

109. RL contains triazole, the conjugate according to any one of claims 79 to 108.

110. RL is 【Chemical 76】 selected from the group consisting of, the conjugate according to any one of claims 79 to 108.

111. RL is 【Chemical 77】 is; 【Chemical 78】 represents a bond to the remainder of the compound, the conjugate according to claim 110.

112. L ２ is -(CR ａ R ｂ ) ａ -POLY １ - and is the conjugate according to any one of claims 79 to 103 and 109 to 111.

113. POLY １ - is a divalent residue of polyethylene glycol (PEG), poly(propylene glycol) (PPG), or a copolymer of ethylene glycol and propylene glycol, the conjugate according to claim 112.

114. The divalent hydrophilic polymer residue has the formula 【Chemical Formula 79】 having R ５ The conjugate according to claim 113, wherein R is hydrogen or methyl and x is an integer between 1 and 100 including both ends.

115. x is 1 to 25 including both ends, the conjugate according to claim 114.

116. x is 13, the conjugate according to claim 115.

117. L ２ is 【Chemical 80】 is, 【Chemical 81】 represents a bond to the remainder of the compound, the conjugate according to any one of claims 79 to 103 and 109 to 111.

118. POLY ２ The conjugate according to claim 117, wherein POLY is a residue of polyethylene glycol (PEG), methoxypolyethylene glycol (mPEG), poly(propylene glycol) (PPG), or a copolymer of ethylene glycol and propylene glycol.

119. POLY ２ is 【Chemical 82】 and R ５ is hydrogen or methyl, and x is an integer between 1 and 100 including both ends, the conjugate according to claim 118.

120. x is 1 to 25 including both ends, the conjugate according to claim 119.

121. x is 11, the conjugate according to claim 120.

122. L ３ The conjugate according to any one of claims 79 to 121, wherein L is -C(O)-AA-.

123. L ３ is -C(O)-AA-Z-(CR ａ R ｂ ) ａ -Z-(CR ａ R ｂ ) ａ -C(O)-, -Z-AA-, or -AA-, the conjugate according to any one of claims 79 to 121.

124. L ３ The conjugate according to any one of claims 79 to 121, wherein L is -AA-.

125. -AA- is 【Chemical 83】 is, the conjugate according to claim 124.

126. The conjugate according to any one of claims 122 to 124, wherein AA is a dipeptide residue, a tripeptide residue, a tetrapeptide residue, or a pentapeptide residue.

127. The conjugate according to claim 126, wherein AA is a tripeptide residue.

128. The conjugate according to any one of claims 122 to 124 and 126 to 127, wherein AA comprises at least one amino acid residue selected from alanine, glycine, valine, and asparagine.

129. The conjugate according to any one of claims 122 to 124 and 126 to 127, wherein AA comprises at least one amino acid residue selected from alanine and glycine.

130. AA is 【Chemical 84】 The conjugate according to any one of claims 122 to 124 and 126 to 127, which is selected from the group consisting of

131. L ３ is -C(O)-Z-(CR ａ R ｂ ) ａ -C(O)-Z-L ４ -OC(O)-; L ４ is 【Chemical 85】 ; The conjugate according to any one of claims 79 to 121, wherein Su is a hexose form of a monosaccharide.

132. Su is 【Chemical 86】 The conjugate according to claim 131.

133. Su is 【Chemical 87】 The conjugate according to claim 132.

134. The conjugate according to any one of claims 79 to 133, wherein D is a cytotoxic payload selected from a tubulin inhibitor, a DNA damaging agent, a DNA topoisomerase I inhibitor, a DNA topoisomerase II inhibitor, and an RNA polymerase II inhibitor.

135. The conjugate according to claim 134, wherein D is a cytotoxic payload selected from the group consisting of hemiasterlin or a derivative thereof, camptothecin or a derivative thereof, camptothexin, anthracycline or a derivative thereof, PNU-159682 or a derivative thereof, PBD or a derivative thereof, and duocarmycin or a derivative thereof.

136. D is 【Chemical 88】 selected from 【Chemical 89】 which represents the bond to the remainder of the compound. The conjugate according to claim 134.

137. The conjugate according to claim 134, wherein D is an exatecan payload.

138. 【Chemical Formula 90-1】 【Chemical Formula 90-2】 【Chemical Formula 90-3】 【Chemical Formula 90-4】 【Chemical Formula 90-5】 【Chemical Formula 90-6】 【Chemical Formula 90-7】 【Chemical Formula 90-8】 【Chemical Formula 90-9】 【Chemical Formula 90-10】 【Chemical Formula 90-11】 【Chemical Formula 90-12】 【Chemical Formula 90-13】 【Chemical Formula 90-14】 【Chemical Formula 90-15】 Or a pharmaceutically acceptable salt thereof. The conjugate according to claim 79.

139. Formula: 【Chemical Formula 91-1】 【Chemical Formula 91-2】 Or a pharmaceutically acceptable salt thereof. The conjugate according to claim 138.

140. Formula: 【Chemical Formula 92】 The conjugate according to claim 80, or a pharmaceutically acceptable salt thereof.

141. A pharmaceutical composition comprising a compound according to any one of claims 1 to 77 or a pharmaceutically acceptable salt thereof, or a conjugate according to any one of claims 78 to 140 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient, carrier, or diluent.

142. A method of treating a disease or disorder in a subject in need thereof, the method comprising administering a compound according to any one of claims 1 to 77 or a pharmaceutically acceptable salt thereof, a conjugate according to any one of claims 78 to 140 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 141.

143. A method of inhibiting tubulin polymerization in a subject in need thereof, the method comprising administering a compound according to any one of claims 1 to 77 or a pharmaceutically acceptable salt thereof, a conjugate according to any one of claims 78 to 140 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 141.

144. The method according to claim 142, wherein the disease or disorder is abnormal cell proliferation.

145. The method according to claim 144, wherein the abnormal cell proliferation is cancer.

146. The method according to claim 145, wherein the cancer is small cell lung cancer, non-small cell lung cancer, ovarian cancer, platinum-resistant ovarian cancer, ovarian adenocarcinoma, endometrial cancer, breast cancer, breast cancer overexpressing HER2, triple-negative breast cancer, lymphoma, large cell lymphoma, diffuse mixed histiocytic and lymphocytic lymphoma, follicular B cell lymphoma, colon cancer, colorectal cancer, colon adenocarcinoma, colorectal adenocarcinoma, melanoma, prostate cancer, or multiple myeloma.

147. Use of a compound according to any one of claims 1 to 77 or a pharmaceutically acceptable salt thereof, a conjugate according to any one of claims 78 to 140 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 141, for treating a disease or disorder in a subject in need thereof.

148. Use of a compound according to any one of claims 1 to 77 or a pharmaceutically acceptable salt thereof, a conjugate according to any one of claims 78 to 140 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 141, for preparing a medicament for treating a disease or disorder in a subject in need thereof.

149. Use according to claim 147 or 148, wherein the treatment is inhibition of tubulin polymerization. Use of a compound according to any one of claims 1 to 77 or a pharmaceutically acceptable salt thereof, a conjugate according to any one of claims 78 to 140 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 141, for preparing a medicament for inhibiting tubulin polymerization in a subject in need thereof.

150. Use according to claim 147 or 148, wherein the disease or disorder is abnormal cell proliferation or cancer.

151. A method of reducing cell proliferation in a subject in need thereof, the method comprising administering a compound according to any one of claims 1 to 77 or a pharmaceutically acceptable salt thereof, a conjugate according to any one of claims 78 to 140 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 141.

152. A method of generating a conjugate, the method comprising contacting a compound according to any one of claims 1 to 77 with a second compound under conditions suitable for conjugating the compound according to any one of claims 1 to 77 with the second compound, wherein the second compound comprises an alkyne, cyclooctyne, strained alkene, tetrazine, methylcyclopropene, thiol, maleimide, carbonyl, amine, oxyamine, or azide.

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