β-Glucuronide linker-payload, its protein conjugate, and its method

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 under 35 U.S.C. § 119 of U.S. Provisional Patent Application No. 63 / 355,975, filed on June 27, 2022, the entire content of which is incorporated herein by reference.

[0002]

[0002] Provided herein are β - glucuronide linker - payload compounds, and their polymeric conjugates; pharmaceutical compositions comprising β - glucuronide linker - payload compounds and / or conjugates; methods of making β - glucuronide linker - payload compounds and / or conjugates; and methods of using β - glucuronide linker - payload compounds, conjugates, and compositions for treatment. The β - glucuronide linker - payload compounds, conjugates, and compositions are useful, for example, in methods of treating and preventing cell proliferation and cancer, methods of detecting cell proliferation and cancer, and methods of diagnosing cell proliferation and cancer.

Background Art

[0003]

[0003] Biopharmaceuticals offer abundant treatment and diagnostic possibilities for patients worldwide. However, many drugs based on macromolecules such as proteins, peptides, and antibodies pose 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] Compounds of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), (IVB), and their sub-formulas, compositions containing such compounds, methods of making such compounds, and methods of using such compounds, conjugates, and compositions in treatment and diagnosis are provided herein. The compounds of the present disclosure, including those of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), (IVB), and their sub-formulas and embodiments, are useful for modulating the bioavailability and ADME of macromolecular compounds. In certain embodiments, the compounds can be used to prepare prodrug conjugates of macromolecular compounds for in vivo or other uses. In certain embodiments, the compounds and conjugates are characterized by functional groups suitable for enzymatic cleavage to release payload compounds for in vivo or other uses. 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 macromolecules in vivo is provided.

[0005]

[0005] In some embodiments, the compound of formula (I)

[0006]

Chem.

[0007]

[0006] In some embodiments, the compound of formula (I)

[0008]

Chemical formula

[0009]

[0007] In some embodiments, the present disclosure provides a compound having the structure of formula (III):

[0010]

Chemical Formula

[0011]

[0008] In some embodiments,

[0012]

Chemical formula

[0013]

[0009] In some embodiments,

[0014]

Chemical formula

[0015]

[0010] In some embodiments, conjugates are provided that include residues of compounds of formula (I), (IA), (IB), (III), (IIIA), (IIIB), or pharmaceutically acceptable salts thereof, linked to a second compound.

[0016]

[0011] In some embodiments, the following chemical structures

[0017]

Chemical formula

[0018]

Chemical formula

[0019]

[0012] In some embodiments, the following chemical structures

[0020]

Chemical formula

[0021]

Chemical formula

[0022]

[0013] In certain embodiments, pharmaceutical compositions are provided that comprise a compound of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), or (IVB) and a pharmaceutically acceptable excipient, carrier, or diluent. Any suitable pharmaceutical composition may be used.

[0023]

[0014] In certain embodiments, the compounds of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), and (IVB), or pharmaceutical compositions thereof, are useful for treatment. In certain embodiments, the compounds of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), and (IVB), or pharmaceutical compositions thereof, are useful for the treatment of cancer. In certain embodiments, the compounds of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), and (IVB), or pharmaceutical compositions thereof, are useful in medicine.

[0024]

[0015] In certain embodiments, a method of inhibiting tubulin polymerization in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), or (IVB), or a pharmaceutical composition thereof, is provided. In certain embodiments, a method of decreasing cell proliferation in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), or (IVB), or a pharmaceutical composition thereof, is provided. In certain embodiments, a method of treating cell proliferation or cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), or (IVB), or a pharmaceutical composition thereof, is provided.

[0025]

[0016] In certain embodiments, a method of generating a conjugate, the method comprising contacting a compound of formula (I), (IA), (IB), (III), (IIIA), or (IIIB) with a second compound under conditions suitable for conjugating the compound of formula (I), (IA), (IB), (III), (IIIA), or (IIIB) to the second compound, wherein the second compound comprises an alkyne, cyclooctyne, strained alkene, tetrazine, methylcyclopropene, thiol, maleimide, carbonyl, amine, oxyamine, or azide, is provided.

Brief Description of the Drawings

[0026]

Figure 1

[0017] A diagram showing MALDI-ToF data for the drug-antibody ratio (DAR) after conjugation with an antibody.

Figure 2A

Figure 2B

Figure 2C

Figure 2D

Figure 2E

Figure 2F

Figure 3A

Figure 3B

Figure 3C

Figure 3D

[0027]

[0020] Compounds of formulas (I), (IA), (IB), (III), (IIIA), and (IIIB), and conjugates of formulas (II), (IIA), (IIB), (IV), (IVA), and (IVB) that are useful for modulating the bioavailability and ADME of such polymeric conjugate compounds are described herein. In some examples, the compounds described herein are useful for preparing conjugates of polymers for in vivo use, such as prodrugs.

[0028]

[0021] Definitions Unless otherwise defined, all technical terms, notations, and other scientific terms used herein are intended to have the meaning commonly understood by one of ordinary skill in the art to which this disclosure pertains. In some examples, terms having commonly understood meanings are defined herein 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, the widely utilized molecular cloning methodologies. 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.

[0029]

[0022] As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.

[0030] As used herein, the term "about" refers to the indicated value as well as ranges above and below that value, including these. 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.

[0031]

[0024] When referring to the compounds provided herein, unless otherwise indicated, the following terms have the following meanings. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. If there are multiple definitions for a term herein, the definitions in this section shall prevail unless otherwise expressly stated.

[0032]

[0025] "Alkoxy" and "alkoxyl" refer to an -OR’’ group, where 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.

[0033]

[0026] As used herein, the term "alkoxyamine" refers to an -alkylene-O-NH2 group, where 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.

[0034]

[0027] As used herein, the term "alkyl" refers to saturated straight-chain or branched hydrocarbons, unless otherwise specified. In certain embodiments, the alkyl group is a primary, secondary, or tertiary hydrocarbon. In certain embodiments, the alkyl group contains from 1 to 10 carbon atoms (i.e., C 1- C 10 alkyl). In certain embodiments, alkyl is lower alkyl, such as C 1-6 alkyl and the like. 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 alkyl substituted with one, three, or three groups independently selected from, for example, 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, alkyl is unsubstituted.

[0035]

[0028] 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, alkylene is unsubstituted.

[0036]

[0029] "Alkenyl", in certain embodiments, refers to an olefinically unsaturated hydrocarbon group that can be linear or branched, has up to about 11 carbon atoms or 2 to 6 carbon atoms (e.g., "lower alkyl"), and has at least one or one to two olefinically unsaturated sites. "Substituted alkenyl" refers to an alkenyl group substituted as described herein for alkyl.

[0037]

[0030] "Alkenylene" refers to a divalent alkenyl as defined herein. Lower alkenylene is, for example, C2-C6-alkenylene.

[0038]

[0031] "Alkynyl", in certain embodiments, refers to an acetylenically unsaturated hydrocarbon group that can be linear or branched, has up to about 11 carbon atoms or 2 to 6 carbon atoms (e.g., "lower alkynyl"), and has at least one or one to two acetylenically unsaturated 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.

[0039]

[0032] "Alkynylene" refers to a divalent alkynyl as defined herein. Lower alkynylene is, for example, C2-C6-alkynylene.

[0040]

[0033] "Amino" refers to -NH2.

[0041]

[0034] The term "alkylamino", as used herein, unless otherwise specified, refers to an -NHR'' group (wherein R'' is, for example, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, C 1-10 haloalkyl, etc.). In certain embodiments, alkylamino is C 1-6It is alkylamino.

[0042]

[0035] As used herein, the term "dialkylamino", unless otherwise specified, refers to an -NR''R'' group (wherein each R'' is independently a C 1-10 alkyl as defined herein). In certain embodiments, the dialkylamino is, for example, di-C 1-6 alkylamino, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, C 1-10 haloalkyl, and the like.

[0043]

[0036] As used herein, the term "aryl", unless otherwise specified, refers to phenyl, biphenyl, or naphthyl. This term includes both substituted and unsubstituted moieties. The 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, where each moiety is 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 the arylamino and aryloxy substituents is further unsubstituted.

[0044]

[0037] 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).

[0045]

[0038] As used herein, the term "arylene", unless otherwise specified, refers to a divalent aryl group as defined herein.

[0046]

[0039] As used herein, the term "aryloxy", unless otherwise specified, refers to an -OR group (wherein R is an aryl as defined herein).

[0047]

[0040] "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.

[0048]

[0041] "Aralkylene" refers to a -CH2-arylene-, -arylene-CH2-, or -CH2-arylene-CH2- group (wherein arylene is as defined herein). "Substituted aralkylene" refers to an aralkylene as defined herein, wherein the aralkylene group is substituted as defined herein for aryl.

[0049]

[0042] "Carboxyl" or "carboxy" refers to -C(O)OH or -COOH.

[0050]

[0043] As used herein, the term "cycloalkyl", unless otherwise specified, refers to a saturated cyclic hydrocarbon. In certain embodiments, the cycloalkyl group can be a saturated, and / or bridged, and / or unbridged, and / or fused bicyclic group. In certain embodiments, the cycloalkyl group contains from 3 to 10 carbon atoms (i.e., C3-C 10 cycloalkyl). In some embodiments, cycloalkyl is from 3 to 15 carbons (C 3-15 ), from 3 to 10 carbons (C 3-10 ), from 3 to 7 carbons (C 3-7) or having 3 to 6 carbons (C 3-6 ), i.e., "lower cycloalkyl". In certain embodiments, the cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexylmethyl, cycloheptyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, decalinyl, or adamantyl.

[0051]

[0044] As used herein, the term "carbocyclic ring" refers to a saturated ring, unsaturated ring, or aromatic ring in which each atom of the ring is carbon, unless otherwise specified. In certain embodiments, the carbocyclic group can be a saturated and / or bridged and / or unbridged and / or fused bicyclic group, and / or a spiro bicyclic group. In some embodiments, the carbocyclic ring includes a 3- to 10-membered monocyclic ring, a 6- to 12-membered bicyclic ring, and / or a 6- to 12-membered bridged ring. In some embodiments, each ring of the bicyclic carbocyclic ring can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. In some embodiments, an aromatic ring, for example, phenyl, may be fused to a saturated ring or an unsaturated ring, such as cyclohexane, cyclopentane, or cyclohexene. The bicyclic carbocyclic ring includes any combination of saturated, unsaturated, and aromatic bicyclic rings as permitted by valence. The bicyclic carbocyclic ring 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 carbocyclic group includes 3 to 10 carbon atoms (i.e., C3-C 10 carbocyclic ring). In certain embodiments, the carbocyclic group includes 3 to 12 carbon atoms (i.e., C3-C 12 carbocyclic ring). In some embodiments, the carbocyclic ring has 3 to 15 carbons (C 3-15 ), 3 to 12 carbons (C 3-12 ), 3 to 10 carbons (C 3-10 ), 3 to 7 carbons (C 3-7) or has 3 to 6 carbons (C3 - C6). In certain embodiments, the carbocyclic group is cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cyclohexylmethyl, cycloheptyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, decalinyl, phenyl, indanyl, naphthyl, or adamantyl.

[0052]

[0045] As used herein, the term "cycloalkylene" refers to a divalent cycloalkyl group as defined herein. In certain embodiments, the cycloalkylene group is cyclopropylene

[0053]

Chem.

[0054]

Chem.

[0055]

Chem.

[0056]

Chem.

[0057]

Chem.

[0058] As used herein, the term "cycloalkylalkyl" refers to an alkyl group as defined herein substituted with one or two cycloalkyl groups as defined herein, unless otherwise specified.

[0059] As used herein, the term "ester" refers to -C(O)OR or -COOR, where R is alkyl as defined herein.

[0060] As used herein, the term "fluorene" may be substituted with one or more chemical functional groups described herein, where any one or more carbons having one or more hydrogens.

[0061] [Chemical formula] refers to.

[0062] As used herein, the term "haloalkyl" refers to an alkyl group as defined herein substituted with one or more halogen atoms (e.g., in some embodiments, 1, 2, 3, 4, or 5), independently selected.

[0063] 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.

[0064] 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.

[0065]

[0052] The term "heterocycloalkyl" refers to a monovalent monocyclic or polycyclic non-aromatic ring system in which one or more of the ring atoms are heteroatoms independently selected from oxygen (O), sulfur (S), and nitrogen (N) (e.g., 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. In certain embodiments, heterocycloalkyl is a monovalent monocyclic or polycyclic fully saturated ring system. In certain embodiments, the heterocycloalkyl 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 can be attached to the core structure at any heteroatom or carbon atom that results in the creation of a stable compound. In certain embodiments, heterocycloalkyl may include a fused ring system or a bridged 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 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, thiamorpholinyl, thiazolidinyl, tetrahydroquinolinyl, and 1,3,5-trithianyl. In certain embodiments, heterocycloalkyl may also be optionally substituted as described herein.In certain embodiments, the heterocycloalkyl 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 group may contain one, two, three, or four heteroatoms. One of ordinary skill in the art will recognize that a 4-membered heterocycloalkyl may generally contain one or two heteroatoms, a 5- or 6-membered heterocycloalkyl may generally contain one, two, or three heteroatoms, and a 7- to 10-membered heterocycloalkyl may generally contain one, two, three, or four heteroatoms.

[0066] As used herein, the term "heterocyclic ring" refers to a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include nitrogen (N), oxygen (O), silicon (Si), phosphorus (P), boron (B), and sulfur (S) atoms, with nitrogen or sulfur atoms optionally oxidized and nitrogen atoms optionally quaternized, and the remaining ring atoms of non-aromatic rings being carbon atoms. Heterocyclic rings include 3- to 10-membered monocyclic rings, 3- to 12-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. In certain embodiments, the heterocyclic ring is a monocyclic or polycyclic fully saturated ring system of valence one. The bicyclic heterocyclic ring includes any combination of saturated, unsaturated, and aromatic bicyclic rings as permitted by valence. In some embodiments, an aromatic ring, such as pyridyl, may be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, morpholine, piperidine, or cyclohexene. The bicyclic heterocyclic ring 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, a heterocycloalkyl or "heterocyclic ring" group can be a saturated, and / or bridged, and / or unbridged, and / or fused bicyclic group, and / or a spiro ring bicyclic group. The term "unsaturated heterocyclic ring" refers to a heterocyclic ring having at least one degree of unsaturation and excluding aromatic heterocyclic rings. Examples of unsaturated heterocyclic rings include dihydropyrrole, dihydrofuran, oxazoline, pyrazoline, and dihydropyridine.

[0067] As used herein, "heterocycloalkylene" refers to a divalent heterocycloalkyl as defined herein.

[0068] As used herein, the term "heteroaryl" refers to a monocyclic aromatic group and / or a 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 remainder 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.

[0069]

[0056] The term "heteroarylene" as used herein refers to a divalent heteroaryl group as defined herein. "Replaced heteroarylene" is heteroarylene substituted as defined for aryl.

[0070]

[0057] 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 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).

[0071] "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, acid addition salts formed by (1) 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) a metal ion such as an alkali metal ion, an alkaline earth metal ion, or an aluminum ion, or an alkali metal hydroxide or an alkaline earth metal hydroxide 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 an organic base such as an aliphatic, alicyclic, or aromatic organic amine 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.

[0072]

[0059] Pharmaceutically acceptable salts further include, by way of example and without limitation, sodium salts, potassium salts, calcium salts, magnesium salts, ammonium salts, and tetraalkylammonium salts, etc. And when the compound contains a basic functional group, hydrohalic acid salts, 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., salts of non-toxic organic acids or inorganic acids are included.

[0073] The terms “substantially free of” or “in the substantial absence of” with respect to a composition refer to a composition that contains at least 85 wt% or 90 wt%, in certain embodiments 95 wt%, 98 wt%, 99 wt%, or 100 wt%; or in certain embodiments 95%, 98%, 99%, or 100% of a 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 the two enantiomers. In certain embodiments, in the methods and compounds provided herein, the compound is substantially free of one of the 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).

[0074] Similarly, the term “isolated” with respect to a composition refers to a composition that contains at least 85 wt%, 90 wt%, 95 wt%, 98 wt%, or 99 wt% to 100 wt% of a compound, with the balance containing other chemical species, enantiomers, or diastereomers.

[0075] “Solvate” refers to a compound or a salt thereof provided herein that further contains 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.

[0076] “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 present naturally for a given atom. Atoms having 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 expressly indicated, when a position is specifically shown as hydrogen (H), that position is understood to have hydrogen of its natural isotopic composition.

[0077]

[0064] "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.

[0078]

[0065] "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.

[0079]

[0066] As used herein, the "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.

[0080]

[0067] Similarly, as used herein, the "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.

[0081]

[0068] 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 is of a mass of at least 1000 Da. In certain embodiments, the polymer has at least 1000 atoms. In certain embodiments, the polymer may be modified. For example, a protein, peptide, or antibody may be modified with one or more carbohydrates and / or small molecule therapeutic compounds.

[0082]

[0069] 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, for example, Paul, Fundamental Immunology, 7th Edition, Chapter 5 (2013) Lippincott Williams & Wilkins, Philadelphia, PA. Briefly, each heavy chain typically comprises 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 comprises three domains designated C H 1 (or CH1), C H 2 (or CH2), and C H 3 (or CH3). Each light chain typically comprises a light chain variable region (V L or VL) and a light chain constant region. The light chain constant region typically comprises one domain designated C L or CL.

[0083] 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.

[0084] 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 the 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, unnatural (or non-natural) 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, seryl, threonyl, 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-seryl, β-threonyl, 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 constituting 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.

[0085]

[0072] The term "conjugate" refers to a compound or drug moiety described herein that is linked to one or more polymer moieties. The polymer moiety is as defined herein or any polymer that is considered suitable for those skilled 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 polymer moiety via a covalent bond, or the compound or drug moiety can be indirectly linked to the polymer moiety via a linker. Typically, the linker is covalently bonded to the polymer moiety and also covalently bonded to the compound or drug moiety.

[0086]

[0073] "pAMF", "pAMF residue", or "pAMF variant" refers to a mutant phenylalanine residue (i.e., para-azidomethyl-L-phenylalanine) added or substituted to a polypeptide.

[0087] 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 another 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 of forming covalent bonds with more than one compound or drug moiety. After the linker forms 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, considering 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 bound to a polymer, a linker residue covalently bound to a compound or drug moiety, and / or a linker residue covalently bound to a polymer and covalently bound to a compound or drug moiety. In some embodiments, the linker is a cleavable linker. For example, a cleavable linker may be labile in vivo or released by enzymatic function, whether or not manipulated. In some embodiments, the linker is a non-cleavable linker. For example, a non-cleavable linker may be released upon degradation of the polymeric moiety.

[0088]

[0075] 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 maximal manifestation of a particular response induced, evoked, or enhanced by the particular test compound.

[0089]

[0076] 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.

[0090]

[0077] 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 domestic animal (e.g., a horse, a cow, a pig, etc.) or a pet (e.g., a dog or a cat). In certain embodiments, the subject is a human.

[0091]

[0078] As used herein, the terms "therapeutic agent" and "therapeutic agents" refer to any agent or payload 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, a therapeutic agent is a drug that is known to be useful for, or has been used or is currently being used for, the treatment or prevention of a disorder or one or more of its symptoms.

[0092]

[0079] "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 a condition. The "therapeutically effective amount" can vary, in particular, depending on the compound, the disease or disorder and its severity, and the age, weight, etc. of the subject being treated.

[0093] As used herein, "treating" or "treatment" of any disease or disorder, in certain embodiments, refers to ameliorating a disease or disorder present in a subject. In another embodiment, "treating" or "treatment" includes ameliorating at least one physical parameter that may be undetectable by the subject. In yet another embodiment, "treating" or "treatment" includes modulating a 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 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 progression of a disease or disorder or one or more symptoms thereof, or reducing severity of a disease or disorder or one or more symptoms thereof.

[0094] 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 contacted 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 a variety of mechanisms including, but not limited to, internalization, apoptosis, necrosis, and / or effector function-mediated activity of the conjugate, compound, or drug moiety.

[0095] As used herein, the terms "prophylactic agent" and "prophylactic agents" refer to any agent or payload 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 a drug that is known to be useful or has been used or is currently used to prevent or interfere with the onset, development, progression, and / or severity of a disorder.

[0096] As used herein, the phrase "prophylactically effective amount" refers to an amount of a treatment (e.g., a prophylactic agent or payload) that is 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).

[0097] In some of the chemical structures illustrated herein, certain substituents, chemical groups, and atoms are depicted by one or more bent / wavy / curved lines that cross one or more bonds to indicate the atom to which the substituent, chemical group, and atom are attached (e.g.,

[0098]

Chem.

[0099]

Chem.

[0100] [Chem.] In some structures, such as, this curved / wavy / sinuous line indicates the atoms in the polymer, as well as the atoms in the backbone of a conjugate, compound, or drug moiety to which the exemplified chemical entity is attached.

[0101]

[0085] 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 in 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, the group

[0102] [Chem.] (wherein the subscript q is an integer from 0 to 4, and the position of the substituent R 1 is generally described, i.e., at any vertex of the bond line structure, i.e., not directly bonded to a specific ring carbon atom) includes the following non-limiting examples of groups in which the substituent R 1 is bonded to a specific ring carbon atom:

[0103] [Chem.] .

[0104] The term "site-specific" refers to the modification of a polypeptide at a predetermined 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 at that sequence position recombinantly or synthetically. Similarly, a moiety can be "site-specifically" linked to a residue at a specific sequence position of a polypeptide. In certain embodiments, a polypeptide can comprise two or more site-specific modifications.

[0105]

[0087] Compounds of formula (I), (IA), (IB), (III), (IIIA), and (IIIB) Compounds are provided herein that are useful for modulating one or more properties of a polymer. The compounds can be formed as described herein and can be used to form conjugates with one or more polymers. The conjugates can be useful for therapy or diagnosis. In certain embodiments, the therapy is the treatment of cancer or an inflammatory disease or condition.

[0106]

[0088] The embodiments described herein include the recited compounds, as well as their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, tautomers, and / or mixtures.

[0107]

[0089] This disclosure pertains to compounds of the structure of formula (I)

[0108]

Chem.

[0109]

[0090] The present disclosure relates to a compound having the structure of formula (I)

[0110]

Chemical formula

[0111]

[0091] In some embodiments, a compound of formula (IA)

[0112]

Chemical Formula

[0113]

[0092] In some embodiments, a compound of formula (I) is one of formula (IB)

[0114]

Chemical Formula

[0115]

[0093] In some embodiments, L 1 is -C 1-3 alkylene-. In some embodiments, L 1 is -CH2-. In some embodiments, L 1 is -CH2CH2-. In some embodiments, L 1 is -CH2CH2CH2-.

[0116]

[0094] In some embodiments, p is 1. In some embodiments, Y is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n -X 1 - and at least one alkylene of Y is substituted with one or more substituents selected from R 50 In some embodiments, Y is -X 1 -C 2-6 alkenylene-[X 1 -C 2-6 alkenylene] n -X 1 - and at least one alkenylene of Y is substituted with one or more substituents selected from R 50 In some embodiments, Y is -X 1 -C 2-6 alkynylene-[X 1 -C 2-6 alkynylene] n -X 1 - and at least one alkynylene of Y is substituted with one or more substituents selected from R 50 In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0117]

[0095] In some embodiments, p is 1. In certain embodiments, Y is -X 1 -C 1-4 alkylene-[X 1 -C 1-4 alkylene] n -X 1 - and at least one alkylene of Y is substituted with one or more substituents selected from R 50 In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0118]

[0096] In some embodiments, p is 1. In certain embodiments, Y is -X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-X 1 -, and at least one alkylene of Y is substituted with one or more substituents selected from R 50 In certain embodiments, Y is -X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-X 1 -, and at least one alkylene of Y is substituted with one or more substituents selected from R 50 In certain embodiments, Y is -X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-X 1 -, and at least one alkylene of Y is substituted with one or more substituents selected from R 50

[0119]

[0097] In some embodiments, p is 0. In some embodiments, Y is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n -, and at least one alkylene of Y is substituted with one or more substituents selected from R 50 and the alkylene of Y may be substituted with one or more substituents selected from R 51 In some embodiments, Y is -X 1 -C 2-6 alkenylene-[X 1 -C 2-6 alkenylene] n ​- and at least one alkenylene of Y is substituted with one or more substituents selected from R 50 and the alkenylene of Y may be substituted with one or more substituents selected from R 51 . In some embodiments, Y is -X 1 -C 2-6 alkynylene-[X 1 -C 2-6 alkynylene] n - and at least one alkynylene of Y is substituted with one or more substituents selected from R 50 and the alkynylene of Y may be substituted with one or more substituents selected from R 51 . In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0120]

[0098] In some embodiments, p is 0. In some embodiments, Y is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n - and at least one alkylene of Y is substituted with one or more substituents selected from R 50 and the alkylene of Y may be substituted with one or more substituents selected from R 51 . In some embodiments, Y is -X 1 -C 1-4 alkylene-[X 1 -C 1-4 alkylene] n - and at least one alkylene of Y is substituted with one or more substituents selected from R 50 and the alkylene of Y may be substituted with one or more substituents selected from R 51 . In some embodiments, Y is -X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-X 1 -C1-4 is alkylene, and at least one alkylene of Y is R 50 is substituted with one or more substituents selected from R 51 and the alkylene of Y may be substituted with one or more substituents selected from R 51 . In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, R 51 is independently selected from halogen, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)-. In some embodiments, R 51 is halogen. In some embodiments, R 51 is -CN. In some embodiments, R 51 is -NO2. In some embodiments, R 51 is -OH. In some embodiments, R 51 is -NH2. In some embodiments, R 51 is -C(O)NH2. In some embodiments, R

[0121]

[0099] In some embodiments, X 1 and X 2 are independently selected from -N(R 10 ), -C(O)-, and -N(R 10 )C(O)-. In some embodiments, X 1 and X 2 are independently selected from -NH-, -C(O)-, and -N(R 10 )C(O)-. In some embodiments, X 1 and X 2 are independently selected from -C(O)-, and -N(R 10 )C(O)-. In some embodiments, X 1 and X 2 are independently selected from -C(O)-, and -NHC(O)-.

[0122]

[0100] In certain embodiments, R 50 is -C1-6 Alkylene-X 2 -[C 1-6 Alkylene] m -POLY, and each alkylene of R 50 may be substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 haloalkyl. In some embodiments, R 50 is -C 1-4 Alkylene-X 2 -[C 1-4 Alkylene] m -POLY, and each alkylene of R 50 may be substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 haloalkyl. In some embodiments, each alkylene of R 50 may be substituted with one or more substituents selected from halogen, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 haloalkyl. In some embodiments, m is 0. In some embodiments, m is 1.

[0123]

[0101] In a certain specific embodiment, R 50 is -C 2-6 alkenylene-X 2 -[C 2-6 alkenylene] m -POLY, and each alkenylene of R 50 may be substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocyclic ring, 3- to 12-membered heterocyclic ring, and C 1-10 haloalkyl. In some embodiments, m is 0. In some embodiments, m is 1.

[0124]

[0102] In a certain specific embodiment, R 50 is -C 2-6 alkynylene-X 2 -[C 2-6 alkynylene] m -POLY, and each alkynylene of R 50 may be substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocyclic ring, 3- to 12-membered heterocyclic ring, and C 1-10 haloalkyl. In some embodiments, m is 0. In some embodiments, m is 1.

[0125]

[0103] In certain embodiments, POLY is 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), a polysaccharide, poly(α-hydroxy acid), poly(vinyl alcohol), polyphosphazene, polyoxazoline (POZ), poly(N-acryloylmorpholine), polysarcosine, or a combination thereof. In some embodiments, POLY is polyethylene glycol (PEG). In some embodiments, POLY is methoxypolyethylene glycol (mPEG). In some embodiments, POLY is poly(propylene glycol) (PPG). In some embodiments, POLY is a copolymer of ethylene glycol and propylene glycol. In some embodiments, POLY is poly(oxyethylated polyol). In some embodiments, POLY is poly(olefin alcohol). In some embodiments, POLY is poly(vinyl pyrrolidone). In some embodiments, POLY is poly(hydroxyalkyl methacrylamide). In some embodiments, POLY is poly(hydroxyalkyl methacrylate). In some embodiments, POLY is a polysaccharide. In some embodiments, POLY is poly(α-hydroxy acid). In some embodiments, POLY is poly(vinyl alcohol). In some embodiments, POLY is polyphosphazene. In some embodiments, POLY is polyoxazoline (POZ). In some embodiments, POLY is poly(N-acryloylmorpholine). In some embodiments, POLY is polysarcosine. In some embodiments, POLY is a non-peptidic, water-soluble polymer. In certain embodiments, POLY comprises polyethylene glycol (PEG) or methoxypolyethylene glycol (mPEG). In certain embodiments, POLY is

[0126]

Chemical Formula

[0127] [Chemical formula] represents a bond to the remainder of the compound, and n1 is an integer from 1 to 20. In certain embodiments, n1 is an integer between 5 and 15. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n1 is 3. In some embodiments, n1 is 4. In some embodiments, n1 is 5. In some embodiments, n1 is 6. In some embodiments, n1 is 7. In some embodiments, n1 is 8. In some embodiments, n1 is 9. In some embodiments, n1 is 10. In some embodiments, n1 is 11. In some embodiments, n1 is 12. In some embodiments, n1 is 13. In some embodiments, n1 is 14. In some embodiments, n1 is 15. In some embodiments, n1 is 16. In some embodiments, n1 is 17. In some embodiments, n1 is 18. In some embodiments, n1 is 19. In some embodiments, n1 is 20. In some embodiments, n1 is 21. In some embodiments, n1 is 22. In some embodiments, n1 is 23. In some embodiments, n1 is 24. In some embodiments, n1 is 25. In some embodiments, n1 is 26. In some embodiments, n1 is 27. In some embodiments, n1 is 28. In some embodiments, n1 is 29. In some embodiments, n1 is 30.

[0128]

[0104] In certain embodiments, RL comprises an alkyne, cyclooctyne, strained alkene, tetrazine, amine, methylcyclopropene, thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide. In some embodiments, RL comprises an alkyne. In some embodiments, RL comprises cyclooctyne. In some embodiments, RL comprises a strained alkene. In some embodiments, RL comprises tetrazine. In some embodiments, RL comprises an amine. In some embodiments, RL comprises methylcyclopropene. In some embodiments, RL comprises a thiol. In some embodiments, RL comprises a para-acetyl-phenylalanine residue. In some embodiments, RL comprises oxyamine. In some embodiments, RL comprises maleimide. In some embodiments, RL comprises azide. In certain embodiments, RL is,

[0129]

Chemical formula

[0130]

Chemical formula

[0131]

Chemical formula

[0132]

Chemical formula

[0133] [Chemistry] represents a bond to the remainder of the compound. In some embodiments, RL is

[0134] [Chemistry] and

[0135] [Chemistry] represents a bond to the remainder of the compound. In some embodiments, RL is

[0136] [Chemistry] and

[0137] [Chemistry] represents a bond to the remainder of the compound. In some embodiments, RL is

[0138] [Chemistry] and

[0139] [Chemistry] represents a bond to the remainder of the compound. In some embodiments, RL is

[0140] [Chemistry] and

[0141] [Chemistry] represents the bond to the remainder of the compound. In some embodiments, RL is

[0142]

Chemical formula

[0143]

Chemical formula

[0144]

Chemical formula

[0145]

Chemical formula

[0146]

Chemical formula

[0147]

Chemical formula

[0148]

Chemical formula

[0149]

Chemical formula

[0150]

Chemical formula

[0151]

Chemical formula

[0152]

[0105] In some embodiments, Su is a sugar moiety. In some embodiments, Su is a hexose form of a monosaccharide. Su can be a glucuronic acid or mannose residue. In certain embodiments, Su is

[0153]

Chemical formula

[0154]

Chemical formula

[0155] [Chemical formula] and

[0156] [Chemical formula] represents a bond to the remainder of the compound.

[0157]

[0106] In some embodiments, D is an immunomodulatory payload. In some embodiments, the immunomodulatory payload is an agonist of stimulator of interferon genes (STING), Toll-like receptor 7 (TLR7), Toll-like receptor 7 / 8 (TLR7 / 8), or Toll-like receptor 8 (TLR8). In some embodiments, the immunomodulatory payload is an agonist of stimulator of interferon genes (STING). In some embodiments, the immunomodulatory payload is an agonist of Toll-like receptor 7 (TLR7). In some embodiments, the immunomodulatory payload is an agonist of Toll-like receptor 7 / 8 (TLR7 / 8). In some embodiments, the immunomodulatory payload is an agonist of Toll-like receptor 8 (TLR8). In some embodiments, the agonist of STING is selected from the group consisting of small molecule agonists of the STING pathway, antibodies that activate STING activity, recombinant proteins that activate the STING pathway, TTI-10001, DMXAA (ASA404), CDNs, c-di-GMP, 2’3’-cGAMP, MK-1454, ADU-S100 (MIW815), SB11285, ADU-V19, IACS-8779, IACS-8803, IMSA101, non-CDNs, E7766, MK-2118, diABZI, MSA-2, JNJ-‘6196, bacterial vectors, SYNB1891, and STACT (see, e.g., Luo et al. Molecules 2022, 27, 4638, the entire content of which is incorporated herein by reference). In some embodiments, the agonist of STING is a small molecule agonist of the STING pathway. In some embodiments, the agonist of STING is an antibody that activates STING activity. In some embodiments, the agonist of STING is a recombinant protein that activates the STING pathway. In some embodiments, the agonist of STING is TTI-10001. In some embodiments, the agonist of STING is DMXAA (ASA404). In some embodiments, the agonist of STING is a CDN. In some embodiments, the agonist of STING is c-di-GMP. In some embodiments, the agonist of STING is 2’3’-cGAMP. In some embodiments, the agonist of STING is MK-1454.In some embodiments, the agonist of STING is ADU-S100 (MIW815). In some embodiments, the agonist of STING is SB11285. In some embodiments, the agonist of STING is ADU-V19. In some embodiments, the agonist of STING is IACS-8779. In some embodiments, the agonist of STING is IACS-8803. In some embodiments, the agonist of STING is IMSA101. In some embodiments, the agonist of STING is non-CDN. In some embodiments, the agonist of STING is E7766. In some embodiments, the agonist of STING is MK-2118. In some embodiments, the agonist of STING is diABZI. In some embodiments, the agonist of STING is MSA-2. In some embodiments, the agonist of STING is JNJ-‘6196. In some embodiments, the agonist of STING is a bacterial vector. In some embodiments, the agonist of STING is SYNB1891. In some embodiments, the agonist of STING is STACT.

[0158]

[0107] In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is selected from the group consisting of GS-986, PRTX-007, PRX-034, S-34240, MBS-8, and APR-002 (see, e.g., Bhagchandani et al., Advanced Drug Delivery Reviews 2021, 175, 113803, the entire contents of which are incorporated herein by reference). In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is GS-986. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is PRTX-007. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is PRX-034. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is S-34240. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is MBS-8. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is APR-002.

[0159] In some embodiments, an agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is selected from the group consisting of imiquimod (R837), resiquimod (R848), 852-A (PF-4878691), besifimod (GS-9620), AZD8848, motolimod (VTX-2337), selgantolimod (GS-9688), NKTR-262, RG-7854 (RO7020531), DSP-0509, BDB-001, BDC-1001, LHC-165, SHR-2150, JNJ-4964 (TQ-73334), RO-7119929, DN-1508052, VTX-1463, BNT-411 (SC1), APR-003, ALT-702, TRANSCON, VX-001, SNAPvax, R848-HA, SM360320, and GSK2245035 (see, e.g., Bhagchandani et al., Advanced Drug Delivery Reviews 2021, 175, 113803; and Evans et al., ACS Omega 2019, 4, 13, 15665, the entire contents of each of which are incorporated herein by reference). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is imiquimod (R837). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is resiquimod (R848). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is 852-A (PF-4878691). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is besifimod (GS-9620). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is AZD8848. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is motolimod (VTX-2337). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is selgantolimod (GS-9688). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is NKTR-262. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is RG-7854 (RO7020531).In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is DSP-0509. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is BDB-001. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is BDC-1001. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is LHC-165. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is SHR-2150. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is JNJ-4964 (TQ-73334). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is RO-7119929. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is DN-1508052. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is VTX-1463. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is BNT-411 (SC1). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is APR-003. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is ALT-702. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is TRANSCON. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is VX-001. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is SNAPvax. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is R848-HA. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is SM360320. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is GSK2245035.

[0160] In some embodiments, the agonist of Toll-like receptor 8 (TLR8) is selected from the group consisting of SBT-6050, SBT-6290, and the ZM-TLR8 agonist. In some embodiments, the agonist of Toll-like receptor 8 (TLR8) is SBT-6050. In some embodiments, the agonist of Toll-like receptor 8 (TLR8) is SBT-6290. In some embodiments, the agonist of Toll-like receptor 8 (TLR8) is the ZM-TLR8 agonist.

[0161]

[0110] In certain embodiments, D is a cytotoxic agent or a cytotoxic payload. In certain embodiments, D is an alkylating agent or an alkylating payload. In certain embodiments, D is a bifunctional alkylating agent. In some embodiments, D is a bifunctional alkylating agent selected from the group consisting of cyclophosphamide, mechlorethamine, chlorambucil, and melphalan. In some embodiments, D is cyclophosphamide. In some embodiments, D is mechlorethamine. In some embodiments, D is chlorambucil. In some embodiments, D is melphalan. In some embodiments, D is a monofunctional alkylating agent. In some embodiments, D is a monofunctional alkylating agent selected from the group consisting of dacarbazine, nitrosourea, and temozolomide. In some embodiments, D is dacarbazine. In some embodiments, D is nitrosourea. In some embodiments, D is temozolomide. In certain embodiments, D is a cytoskeletal disruptor (e.g., taxane). In some embodiments, D is a cytoskeletal disruptor selected from the group consisting of paclitaxel, docetaxel, abraxane, and taxotere. In some embodiments, D is paclitaxel. In some embodiments, D is docetaxel. In some embodiments, D is abraxane. In some embodiments, D is taxotere. In certain embodiments, D is epothilone. In some embodiments, D is an epothilone selected from the group consisting of epothilone A, epothilone B, epothilone C, epothilone D, and ixabepilone. In some embodiments, D is epothilone A. In some embodiments, D is epothilone B. In some embodiments, D is epothilone C. In some embodiments, D is epothilone D. In some embodiments, D is ixabepilone. In certain embodiments, D is a histone deacetylase inhibitor. In some embodiments, D is a histone deacetylase inhibitor selected from the group consisting of vorinostat and romidepsin. In some embodiments, D is vorinostat. In some embodiments, D is romidepsin. In certain embodiments, D is a kinase inhibitor.In some embodiments, D is a kinase inhibitor selected from the group consisting of bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, and visomib. In some embodiments, D is bortezomib. In some embodiments, D is erlotinib. In some embodiments, D is gefitinib. In some embodiments, D is imatinib. In some embodiments, D is vemurafenib. In some embodiments, D is visomib. In certain embodiments, D is a nucleotide analog and / or a precursor analog. In some embodiments, 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). In some embodiments, D is azacitidine. In some embodiments, D is azathioprine. In some embodiments, D is capecitabine. In some embodiments, D is cytarabine. In some embodiments, D is doxifluridine. In some embodiments, D is fluorouracil. In some embodiments, D is gemcitabine. In some embodiments, D is hydroxyurea. In some embodiments, D is mercaptopurine. In some embodiments, D is methotrexate. In some embodiments, D is tioguanine (formerly thioguanine). In certain embodiments, D is a peptide antibiotic. In some embodiments, D is a peptide antibiotic selected from the group consisting of bleomycin and actinomycin. In some embodiments, D is bleomycin. In some embodiments, D is actinomycin. In certain embodiments, D is a platinum-based agent or a platinum-based payload. In some embodiments, D is a platinum-based agent or a platinum-based payload selected from the group consisting of carboplatin, cisplatin, and oxaliplatin. In some embodiments, D is carboplatin.In some embodiments, D is cisplatin. In some embodiments, D is oxaliplatin. In certain embodiments, D is a retinoid. In some embodiments, D is a retinoid selected from the group consisting of tretinoin, alitretinoin, and bexarotene. In some embodiments, D is tretinoin. In some embodiments, D is alitretinoin. In some embodiments, D is bexarotene. In certain embodiments, D is a vinca alkaloid and its derivatives. In some embodiments, D is a vinca alkaloid and its derivatives selected from the group consisting of vinblastine, vincristine, vindesine, and vinorelbine. In some embodiments, D is vinblastine. In some embodiments, D is vincristine. In some embodiments, D is vindesine. In some embodiments, D is vinorelbine. In certain embodiments, the cytotoxic agent or cytotoxic payload is a tubulin inhibitor, a DNA topoisomerase I inhibitor, or a DNA topoisomerase II inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a tubulin inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase I inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase I inhibitor selected from the group consisting of irinotecan, SN-38, topotecan, and exatecan. In some embodiments, the cytotoxic agent or cytotoxic payload is irinotecan. In some embodiments, the cytotoxic agent or cytotoxic payload is SN-38. In some embodiments, the cytotoxic agent or cytotoxic payload is topotecan. In some embodiments, the cytotoxic agent or cytotoxic payload is exatecan. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase II inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase II inhibitor selected from the group consisting of etoposide, teniposide, and tafluposide. In some embodiments, the cytotoxic agent or cytotoxic payload is etoposide.In some embodiments, the cytotoxic agent or cytotoxic payload is teniposide. In some embodiments, the cytotoxic agent or cytotoxic payload is tafurposide. In certain embodiments, D is selected from the group consisting of hemiasterlin, camptothecin, and anthracyclines. Anthracyclines can include PNU-159682 and EDA PNU-159682 derivatives. In certain embodiments, the anthracycline is selected from the group consisting of daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, and valrubicin. In some embodiments, the anthracycline is daunorubicin. In some embodiments, the anthracycline is doxorubicin. In some embodiments, the anthracycline is epirubicin. In some embodiments, the anthracycline is idarubicin. In some embodiments, the anthracycline is mitoxantrone. In some embodiments, the anthracycline is valrubicin. In some embodiments, D is hemiasterlin. In some embodiments, D is camptothecin. In some embodiments, D is an anthracycline. In some embodiments, D is PNU-159682. In some embodiments, D is an EDA PNU compound. In some embodiments, D is an EDA PNU-159682 derivative. In certain embodiments, D is hemiasterlin, exatecan, PNU-159682, or an EDA PNU-159682 derivative. In some embodiments, D is hemiasterlin. In some embodiments, D is exatecan. In some embodiments, D is PNU-159682. In some embodiments, D is an EDA PNU-159682 compound or derivative.

[0162]

[0111] Representative compounds of the present disclosure, including the compounds of formulas (I), (IA), and (IB), are shown in Table 1.

[0163]

Table 1-1

[0164]

Table 1-2

[0165]

Table 1-3

[0166]

Table 1-4

[0167]

Table 1-5

[0168]

Table 1-6

[0169]

Table 1-7

[0170]

Table 1-8

[0171]

Table 1-9

[0172]

Table 1-10

[0173]

Table 1-11

[0174]

Table 1-12

[0175]

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

[0176]

Chemical formula

[0177]

Chemical formula

[0178]

Chemical formula

[0179]

Chemical formula

[0180]

Chemical formula

[0181]

Chemical formula

[0182]

Chemical formula

[0183]

Chemical formula

[0184]

Chemical formula

[0185]

[0113] In certain embodiments, the compound of formula (I), (IA), or (IB) is

[0186]

Chemical formula

[0187]

Chemical formula

[0188]

Chemical formula

[0189]

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

[0190] [Chemistry]

[0191] [Chemistry]

[0192] [Chemistry] and are selected from any one of them and pharmaceutically acceptable salts thereof. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0193] [Chemistry] or a salt thereof. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0194] [Chemistry] or a salt thereof. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0195] [Chemistry] or a salt thereof. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0196] [Chemistry] or a salt thereof. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0197] [Chemistry] or a salt thereof. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0198]

Chemical formula

[0199]

[0115] In certain embodiments, the compound of formula (I), (IA), or (IB) is

[0200]

Chemical formula

[0201]

Chemical formula

[0202]

Chemical formula

[0203]

Chemical formula

[0204]

Chemical formula

[0205]

Chem.

[0206]

[0116] In some embodiments, the compound of formula (I), (IA), or (IB) is

[0207]

Chem.

[0208]

Chem.

[0209]

Chem.

[0210]

Chem.

[0211]

Chem.

[0212] [Chemical] or a pharmaceutically acceptable salt of any one of them.

[0213]

[0117] In some embodiments, the compound of formula (I), (IA), or (IB) is

[0214] [Chemical] selected from or a pharmaceutically acceptable salt of any one of them. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0215] [Chemical] or a pharmaceutically acceptable salt of any one of them. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0216] [Chemical] or a pharmaceutically acceptable salt of any one of them.

[0217]

[0118] In some embodiments, the compound of formula (I), (IA), or (IB) is

[0218] [Chemical] selected from or a pharmaceutically acceptable salt of any one of them. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0219] [Chemical] or a pharmaceutically acceptable salt of any one of them. In some embodiments, the compound of formula (I), (IA), or (IB) is

[0220]

Chem.

[0221]

[0119] In some aspects, the present disclosure relates to a compound having the structure of formula (III):

[0222]

Chem.

[0223]

[0120] In some embodiments, the compound of formula (IIIA)

[0224]

Chemical formula

[0225]

[0121] In some embodiments, the compound of formula (III) is of formula (IIIB)

[0226]

Chemical formula

[0227]

[0122] In some embodiments, L 1 is -C 1-3is alkylene. In some embodiments, L 1 is -CH2-. In some embodiments, L 1 is -CH2CH2-. In some embodiments, L 1 is -CH2CH2CH2-.

[0228]

[0123] In some embodiments, Z is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n -X 1 -, -X 1 -C 2-6 alkenylene-[X 1 -C 2-6 alkenylene] n -X 1 -, -X 1 -C 2-6 alkynylene-[X 1 -C 2-6 alkynylene] n -X 1 -, and at least one alkylene, alkenylene, or alkynylene of Z is substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocyclic ring, 3- to 12-membered heterocyclic ring, and C 1-10 haloalkyl. In some embodiments, Z is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n -X 1 -, and the alkylene of Z is halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10)2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocyclic ring, 3- to 12-membered heterocyclic ring, and C 1-10 may be substituted with one or more substituents selected from haloalkyl. In some embodiments, Z is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n -X 1 -.

[0229]

[0124] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0230]

[0125] In some embodiments, X 1 and X 2 are independently selected from -C(O)- and -N(R 10 )C(O)-.

[0231]

[0126] In certain embodiments, RL comprises an alkyne, cyclooctyne, strained alkene, tetrazine, amine, thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide. In some embodiments, RL comprises an alkyne. In some embodiments, RL comprises cyclooctyne. In some embodiments, RL comprises a strained alkene. In some embodiments, RL comprises tetrazine. In some embodiments, RL comprises an amine. In some embodiments, RL comprises a thiol. In some embodiments, RL comprises a para-acetyl-phenylalanine residue. In some embodiments, RL comprises an oxyamine. In some embodiments, RL comprises maleimide. In some embodiments, RL comprises an azide. In certain embodiments, RL is

[0232]

Chemical Structure

[0233] [Chemical formula] represents a bond to the remainder of the compound. In some embodiments, RL is

[0234] [Chemical formula] and

[0235] [Chemical formula] represents a bond to the remainder of the compound. In some embodiments, RL is

[0236] [Chemical formula] and

[0237] [Chemical formula] represents a bond to the remainder of the compound. In some embodiments, RL is

[0238] [Chemical formula] and

[0239] [Chemical formula] represents a bond to the remainder of the compound. In some embodiments, RL is

[0240]

Chem.

[0241]

Chem.

[0242]

Chem.

[0243]

Chem.

[0244]

Chem.

[0245]

Chem.

[0246] [Chemical formula] and

[0247] [Chemical formula] represents the bond to the remainder of the compound. In some embodiments, RL is

[0248] [Chemical formula] and

[0249] [Chemical formula] represents the bond to the remainder of the compound. In some embodiments, RL is

[0250] [Chemical formula] and

[0251] [Chemical formula] represents the bond to the remainder of the compound. In some embodiments, RL is

[0252] [Chemical formula] and

[0253] [Chemical formula] represents a bond to the remainder of the compound. In some embodiments, RL is -N3. In some embodiments, RL is -NH2. In some embodiments, RL is methylcyclopropene. In some embodiments, RL is -SH.

[0254]

[0127] In some embodiments, Su is a sugar moiety. In some embodiments, Su is a hexose form of a monosaccharide. Su can be a glucuronic acid or mannose residue. In certain embodiments, Su is

[0255]

Chemical formula

[0256]

Chemical formula

[0257]

Chemical formula

[0258]

Chemical formula

[0259]

[0128] In certain embodiments, CYTO is a cytotoxic agent or cytotoxic payload. In certain embodiments, the cytotoxic agent or cytotoxic payload is a tubulin inhibitor, a DNA topoisomerase I inhibitor, or a DNA topoisomerase II inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a tubulin inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase I inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase I inhibitor selected from the group consisting of irinotecan, SN-38, topotecan, and exatecan. In some embodiments, the cytotoxic agent or cytotoxic payload is irinotecan. In some embodiments, the cytotoxic agent or cytotoxic payload is SN-38. In some embodiments, the cytotoxic agent or cytotoxic payload is topotecan. In some embodiments, the cytotoxic agent or cytotoxic payload is exatecan. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase II inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase II inhibitor selected from the group consisting of etoposide, teniposide, and tafuriposide. In some embodiments, the cytotoxic agent or cytotoxic payload is etoposide. In some embodiments, the cytotoxic agent or cytotoxic payload is teniposide. In some embodiments, the cytotoxic agent or cytotoxic payload is tafuriposide. In certain embodiments, CYTO is selected from the group consisting of hemiasterlin, camptothecin, and anthracyclines. Anthracyclines can include PNU-159682 and EDA PNU-159682 derivatives. In certain embodiments, the anthracycline is selected from the group consisting of daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, and valrubicin. In some embodiments, the anthracycline is daunorubicin. In some embodiments, the anthracycline is doxorubicin. In some embodiments, the anthracycline is epirubicin.In some embodiments, the anthracycline is idarubicin. In some embodiments, the anthracycline is mitoxantrone. In some embodiments, the anthracycline is valrubicin. In some embodiments, CYTO is hemiasterlin. In some embodiments, CYTO is camptothecin. In some embodiments, CYTO is an anthracycline. In some embodiments, CYTO is PNU-159682. In some embodiments, CYTO is an EDA PNU compound. In some embodiments, CYTO is an EDA PNU-159682 derivative. In certain embodiments, CYTO is hemiasterlin, exatecan, PNU-159682, or an EDA PNU-159682 derivative. In some embodiments, CYTO is hemiasterlin. In some embodiments, CYTO is exatecan. In some embodiments, CYTO is PNU-159682. In some embodiments, CYTO is an EDA PNU-159682 compound or derivative. In some embodiments, CYTO is not an immunostimulatory compound.

[0260]

[0129] In some embodiments, the compound is

[0261]

Chem.

[0262]

[0130] In some embodiments, the compound is

[0263]

Chem.

[0264]

[0131] optically active compound In certain embodiments, the compounds provided herein may have several chiral centers, exist in optically active 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, or stereoisomeric form, and / or mixtures thereof. 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 racemic form through recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or chromatographic separation using a chiral stationary phase. Additionally, 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 distinct enantiomers.

[0265]

[0132] 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 individual enantiomers. This technique can be used when crystals of the individual 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 by which partial or complete separation of a 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 synthesis 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 for 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 an excess of the diastereomers of the desired enantiomer, or the kinetic or thermodynamic crystallization of the diastereomers of the desired enantiomer disrupts the equilibrium such that ultimately, in principle, all substances are converted to the crystalline diastereomers of the desired enantiomer. The desired enantiomer is then derived from the diastereomer. viii) Dynamic 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 dynamic conditions; ix) Enantioselective synthesis from non-racemic precursors - a synthesis 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 of contacting a racemate 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.

[0266]

[0133] In some embodiments, provided herein are compositions of the compounds of the present disclosure that comprise a compound of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), or (IVB) that is substantially free of a designated stereoisomer of the compound. In certain embodiments, in the methods and compounds of the present disclosure, the compound is substantially free of other stereoisomers. In some embodiments, the composition comprises a compound that is at least 85 wt%, 90 wt%, 95 wt%, 98 wt%, or 99 wt% - 100 wt% of the compound, and the balance comprises other chemical species or enantiomers. In some embodiments, provided herein are compositions of a compound of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), or (IVB) that is substantially free of a designated enantiomer of the compound. In certain embodiments, in the methods and compounds of the present disclosure, the compound is substantially free of other enantiomers. In some embodiments, the composition comprises a compound that is at least 85 wt%, 90 wt%, 95 wt%, 98 wt%, or 99 wt% - 100 wt% of the compound, and the balance comprises other chemical species or enantiomers.

[0267]

[0134] Isotope-enriched compound Also provided herein are isotopically enriched compounds, including but not limited to, isotopically enriched compounds of formula (I), (IA), (IB), (II), (IIA), (IIB), (III), (IIIA), (IIIB), (IV), (IVA), or (IVB).

[0268]

[0135] 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).

[0269]

[0136] Isotopic enrichment of a drug can be used to, for example, (1) reduce or eliminate undesirable metabolites; (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.

[0270]

[0137] 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, for example, 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)).

[0271]

[0138] 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 extremely large numbers, e.g., up to 50 or more, meaning that the reaction can be 50 times or more slower when deuterium replaces hydrogen.

[0272]

[0139] 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 similar kinetic isotope effects.

[0273]

[0140] The body of an animal expresses various enzymes for the purpose of eliminating foreign substances such as therapeutic agents or payloads from its circulatory system. Examples of such enzymes include cytochrome P450 enzymes ("CYPs") that react with these foreign substances and convert them into more polar intermediates or metabolites for renal excretion, esterases, proteases, reductases, dehydrogenases, and monoamine oxidases. 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. In many drugs, such oxidation is rapid. Thus, these drugs often require multiple or high daily doses.

[0274]

[0141] 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.

[0275]

[0142] Conjugates of formula (II), (IIA), (IIB), (IV), (IVA), (IVB) Conjugates of a polymer and one of the compounds of formula (I), (IA), (IB), (III), (IIIA), or (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 to one or more compounds of formula (I), (IA), (IB), (III), (IIIA), or (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 formula (I), (IA), (IB), (III), (IIIA), or (IIIB).

[0276]

[0143] The linker can be any linker capable of forming at least one bond with the polymer and at least one bond with the compound of formula (I), (IA), (IB), (III), (IIIA), or (IIIB). Useful linkers are described herein, particularly in the following sections and examples.

[0277]

[0144] 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 10 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.

[0278]

[0145] In some embodiments, the conjugate of formula (II)

[0279] [Chemical formula] (wherein, COMP is the residue of the second compound; L 1 is -C 1-6 alkylene-; Y is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n -[X 1 p -, -X 1 -C 2-6 alkenylene-[X 1 -C 2-6 alkenylene] n -[X 1 p -, or -X 1 -C 2-6 alkynylene-[X 1 -C 2-6 alkynylene] n -[X 1 p -, and at least one of the alkylene, alkenylene, or alkynylene of Y is substituted with one or more substituents selected from R 50 ; The alkylene, alkenylene, or alkynylene of Y may be substituted with one or more substituents selected from R 51 ; R 50 is -C 1-6 alkylene-X 2 -[C 1-6 alkylene] m -POLY, -C 2-6 alkenylene-X 2 -[C 2-6 alkenylene] m -POLY, or -C 2-6 alkynylene-X 2 -[C 2-6 alkynylene] m -POLY, and R 50 ​​​Each alkylene, alkenylene, or alkynylene may be substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 haloalkyl; R 51 is independently selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 haloalkyl; X 1 and X 2 are independently selected from -N(R 10 )-, -C(O)-, and -N(R 10 )C(O)-; R 10 is, in each occurrence, independently selected from hydrogen, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 haloalkyl; POLY is a water-soluble polymer; n is an integer selected from 0, 1, 2, and 3; m is an integer selected from 0 and 1; p is an integer selected from 0 and 1; Su is a hexose form of a monosaccharide; D is a drug moiety; RL is a reactive linker group residue) is provided.

[0280]

[0146] In some embodiments, the conjugate of formula (II)

[0281]

Chemical formula

[0282]

[0147] 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.

[0283]

[0148] In some embodiments, the conjugate of formula (IIA)

[0284]

Chemical Formula

[0285]

[0149] In some embodiments, the compound of formula (II) is of formula (IIB)

[0286]

Chemical formula

[0287]

[0150] In some embodiments, L 1 is -C 1-3 alkylene-. In some embodiments, L 1 is -CH2-. In some embodiments, L 1 is -CH2CH2-. In some embodiments, L 1 is -CH2CH2CH2-.

[0288]

[0151] In some embodiments, p is 1. In some embodiments, Y is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n -X 1 -, and at least one alkylene of Y is substituted with one or more substituents selected from R 50 . In some embodiments, Y is -X 1 -C 2-6 alkenylene-[X 1 -C 2-6 alkenylene] n -X 1 -, and at least one alkenylene of Y is substituted with one or more substituents selected from R 50 . In some embodiments, Y is -X 1 -C 2-6 alkynylene-[X 1 -C 2-6 alkynylene] n -X 1- and at least one alkylene of Y is substituted with one or more substituents selected from R 50 In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0289]

[0152] In some embodiments, p is 1. In certain embodiments, Y is -X 1 -C 1-4 alkylene -[X 1 -C 1-4 alkylene] n -X 1 - and at least one alkylene of Y is substituted with one or more substituents selected from R 50 In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0290]

[0153] In some embodiments, p is 1. In certain embodiments, Y is -X 1 -C 1-4 alkylene -X 1 -C 1-4 alkylene -X 1 -C 1-4 alkylene -X 1 -C 1-4 alkylene -X 1 - and at least one alkylene of Y is substituted with one or more substituents selected from R 50 In certain embodiments, Y is -X 1 -C 1-4 alkylene -X 1 -C 1-4 alkylene -X 1 -C 1-4 alkylene -X 1 - and at least one alkylene of Y is substituted with one or more substituents selected from R 50 In certain embodiments, Y is -X 1 -C 1-4 alkylene -X 1 -C1-4 Alkylene-X 1 - and at least one alkylene of Y is substituted with one or more substituents selected from R 50

[0291]

[0154] In some embodiments, p is 0. In some embodiments, Y is -X 1 -C 1-6 Alkylene-[X 1 -C 1-6 Alkylene] n - and at least one alkylene of Y is substituted with one or more substituents selected from R 50 and the alkylene of Y may be substituted with one or more substituents selected from R 51 In some embodiments, Y is -X 1 -C 2-6 Alkenylene-[X 1 -C 2-6 Alkenylene] n - and at least one alkenylene of Y is substituted with one or more substituents selected from R 50 and the alkenylene of Y may be substituted with one or more substituents selected from R 51 In some embodiments, Y is -X 1 -C 2-6 Alkynylene-[X 1 -C 2-6 Alkynylene] n - and at least one alkynylene of Y is substituted with one or more substituents selected from R 50 and the alkynylene of Y may be substituted with one or more substituents selected from R 51 In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0292]

[0155] In some embodiments, p is 0. In some embodiments, Y is -X 1 -C 1-6 Alkylene-[X 1 -C​1-6 alkylene n -, and at least one alkylene of Y is substituted with one or more substituents selected from R 50 and the alkylene of Y may be substituted with one or more substituents selected from R 51 . In some embodiments, Y is -X 1 -C 1-4 alkylene-[X 1 -C 1-4 alkylene n -, and at least one alkylene of Y is substituted with one or more substituents selected from R 50 and the alkylene of Y may be substituted with one or more substituents selected from R 51 . In some embodiments, Y is -X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-X 1 -C 1-4 alkylene-, and at least one alkylene of Y is substituted with one or more substituents selected from R 50 and the alkylene of Y may be substituted with one or more substituents selected from R 51 . In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, R 51 is independently selected from halogen, -CN, -NO2, -OH, -NH2, -C(O)NH2, and -C(O)-. In some embodiments, R 51 is halogen. In some embodiments, R 51 is -CN. In some embodiments, R 51 is -NO2. In some embodiments, R 51 is -OH. In some embodiments, R 51 is -NH2. In some embodiments, R 51 is -C(O)NH2. In some embodiments, R 51 is -C(O)-.

[0293]

[0156] In some embodiments, X 1 and X 2 are independently selected from -N(R 10 )-, -C(O)-, and -N(R 10 )C(O)-. In some embodiments, X 1 and X 2 are independently selected from -NH-, -C(O)-, and -N(R 10 )C(O)-. In some embodiments, X 1 and X 2 are independently selected from -C(O)- and -N(R 10 )C(O)-. In some embodiments, X 1 and X 2 are independently selected from -C(O)- and -NHC(O)-.

[0294]

[0157] In certain embodiments, R 50 is -C 1-6 alkylene-X 2 -[C 1-6 alkylene] m -POLY, and each alkylene of R 50 is optionally substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 haloalkyl. In some embodiments, R 50 is -C 1-4 alkylene-X 2 -[C 1-4 alkylene] m -POLY, and each alkylene of R 50 is optionally substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10)2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 may be substituted with one or more substituents selected from haloalkyl. In some embodiments, R 50 each alkylene of is halogen, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 may be substituted with one or more substituents selected from haloalkyl. In some embodiments, m is 0. In some embodiments, m is 1.

[0295]

[0158] In certain embodiments, R 50 is -C 2-6 alkenylene-X 2 -[C 2-6 alkenylene] m -POLY, and each alkenylene of R 50 is halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 may be substituted with one or more substituents selected from haloalkyl. In some embodiments, m is 0. In some embodiments, m is 1.

[0296]

[0159] In certain embodiments, R 50 is -C 2-6Alkynylene-X 2 -[C 2-6 alkynylene] m -POLY, and each alkynylene of R 50 is optionally substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocycle, 3- to 12-membered heterocycle, and C 1-10 haloalkyl. In some embodiments, m is 0. In some embodiments, m is 1.

[0297]

[0160] In certain embodiments, POLY is 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), a polysaccharide, poly(α-hydroxy acid), poly(vinyl alcohol), polyphosphazene, polyoxazoline (POZ), poly(N-acryloylmorpholine), polysarcosine, or a combination thereof. In some embodiments, POLY is polyethylene glycol (PEG). In some embodiments, POLY is methoxypolyethylene glycol (mPEG). In some embodiments, POLY is poly(propylene glycol) (PPG). In some embodiments, POLY is a copolymer of ethylene glycol and propylene glycol. In some embodiments, POLY is poly(oxyethylated polyol). In some embodiments, POLY is poly(olefin alcohol). In some embodiments, POLY is poly(vinyl pyrrolidone). In some embodiments, POLY is poly(hydroxyalkyl methacrylamide). In some embodiments, POLY is poly(hydroxyalkyl methacrylate). In some embodiments, POLY is a polysaccharide. In some embodiments, POLY is poly(α-hydroxy acid). In some embodiments, POLY is poly(vinyl alcohol). In some embodiments, POLY is polyphosphazene. In some embodiments, POLY is polyoxazoline (POZ). In some embodiments, POLY is poly(N-acryloylmorpholine). In some embodiments, POLY is polysarcosine. In some embodiments, POLY is a non-peptidic, water-soluble polymer. In certain embodiments, POLY comprises polyethylene glycol (PEG) or methoxypolyethylene glycol (mPEG). In certain embodiments, POLY is

[0298]

Chemical formula

[0299] [Chem.] represents a bond to the remainder of the compound, and n1 is an integer from 1 to 20. In certain embodiments, n1 is an integer between 5 and 15. In some embodiments, n1 is 1. In some embodiments, n1 is 2. In some embodiments, n1 is 3. In some embodiments, n1 is 4. In some embodiments, n1 is 5. In some embodiments, n1 is 6. In some embodiments, n1 is 7. In some embodiments, n1 is 8. In some embodiments, n1 is 9. In some embodiments, n1 is 10. In some embodiments, n1 is 11. In some embodiments, n1 is 12. In some embodiments, n1 is 13. In some embodiments, n1 is 14. In some embodiments, n1 is 15. In some embodiments, n1 is 16. In some embodiments, n1 is 17. In some embodiments, n1 is 18. In some embodiments, n1 is 19. In some embodiments, n1 is 20. In some embodiments, n1 is 21. In some embodiments, n1 is 22. In some embodiments, n1 is 23. In some embodiments, n1 is 24. In some embodiments, n1 is 25. In some embodiments, n1 is 26. In some embodiments, n1 is 27. In some embodiments, n1 is 28. In some embodiments, n1 is 29. In some embodiments, n1 is 30.

[0300]

[0161] In certain embodiments, RL comprises an alkyne, cyclooctyne, strained alkene, tetrazine, amine, methylcyclopropene, thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide. In some embodiments, RL comprises an alkyne. In some embodiments, RL comprises cyclooctyne. In some embodiments, RL comprises a strained alkene. In some embodiments, RL comprises tetrazine. In some embodiments, RL comprises an amine. In some embodiments, RL comprises methylcyclopropene. In some embodiments, RL comprises a thiol. In some embodiments, RL comprises a para-acetyl-phenylalanine residue. In some embodiments, RL comprises oxyamine. In some embodiments, RL comprises maleimide. In some embodiments, RL comprises azide. In certain embodiments, RL is selected from the group consisting of

[0301]

Chem.

[0302]

Chem.

[0303]

Chem.

[0304]

Chem.

[0305]

Chem.

[0306]

Chem.

[0307]

Chem.

[0308]

Chem.

[0309]

Chem.

[0310]

Chem.

[0311]

Chem.

[0312]

Chem.

[0313] [Chem.] represents a bond to the remainder of the compound. In some embodiments, RL is

[0314] [Chem.] and

[0315] [Chem.] represents a bond to the remainder of the compound. In some embodiments, RL is

[0316] [Chem.] and

[0317] [Chem.] represents a bond to the remainder of the compound. In some embodiments, RL is

[0318] [Chem.] and

[0319] [Chem.] represents a bond to the remainder of the compound. In some embodiments, RL is

[0320] [Chem.] and R Tis C 1-6 is alkyl,

[0321]

Chem.

[0322]

Chem.

[0323]

Chem.

[0324]

Chem.

[0325]

Chem.

[0326]

Chem.

[0327]

Chem.

[0328]

Chem.

[0329]

Chem.

[0330]

[0162] In some embodiments, Su is a sugar moiety. In some embodiments, Su is a hexose form of a monosaccharide. Su can be a glucuronic acid or mannose residue. In certain embodiments, Su is

[0331]

Chem.

[0332]

Chem.

[0333]

Chem.

[0334]

Chem.

[0335]

[0163] In some embodiments, D is an immunomodulatory payload. In some embodiments, the immunomodulatory payload is an agonist of interferon gene stimulator (STING), Toll-like receptor 7 (TLR7), Toll-like receptor 7 / 8 (TLR7 / 8), or Toll-like receptor 8 (TLR8). In some embodiments, the immunomodulatory payload is an agonist of interferon gene stimulator (STING). In some embodiments, the immunomodulatory payload is an agonist of Toll-like receptor 7 (TLR7). In some embodiments, the immunomodulatory payload is an agonist of Toll-like receptor 7 / 8 (TLR7 / 8). In some embodiments, the immunomodulatory payload is an agonist of Toll-like receptor 8 (TLR8). In some embodiments, the agonist of STING is selected from the group consisting of small molecule agonists of the STING pathway, antibodies that activate STING activity, recombinant proteins that activate the STING pathway, TTI-10001, DMXAA (ASA404), CDNs, c-di-GMP, 2’3’-cGAMP, MK-1454, ADU-S100 (MIW815), SB11285, ADU-V19, IACS-8779, IACS-8803, IMSA101, non-CDNs, E7766, MK-2118, diABZI, MSA-2, JNJ-‘6196, bacterial vectors, SYNB1891, and STACT (see, e.g., Luo et al. Molecules 2022, 27, 4638, the entire content of which is incorporated herein by reference). In some embodiments, the agonist of STING is a small molecule agonist of the STING pathway. In some embodiments, the agonist of STING is an antibody that activates STING activity. In some embodiments, the agonist of STING is a recombinant protein that activates the STING pathway. In some embodiments, the agonist of STING is TTI-10001. In some embodiments, the agonist of STING is DMXAA (ASA404). In some embodiments, the agonist of STING is a CDN. In some embodiments, the agonist of STING is c-di-GMP. In some embodiments, the agonist of STING is 2’3’-cGAMP. In some embodiments, the agonist of STING is MK-1454.In some embodiments, the agonist of STING is ADU-S100 (MIW815). In some embodiments, the agonist of STING is SB11285. In some embodiments, the agonist of STING is ADU-V19. In some embodiments, the agonist of STING is IACS-8779. In some embodiments, the agonist of STING is IACS-8803. In some embodiments, the agonist of STING is IMSA101. In some embodiments, the agonist of STING is non-CDN. In some embodiments, the agonist of STING is E7766. In some embodiments, the agonist of STING is MK-2118. In some embodiments, the agonist of STING is diABZI. In some embodiments, the agonist of STING is MSA-2. In some embodiments, the agonist of STING is JNJ-‘6196. In some embodiments, the agonist of STING is a bacterial vector. In some embodiments, the agonist of STING is SYNB1891. In some embodiments, the agonist of STING is STACT.

[0336]

[0164] In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is selected from the group consisting of GS-986, PRTX-007, PRX-034, S-34240, MBS-8, and APR-002 (see, e.g., Bhagchandani et al. Advanced Drug Delivery Reviews 2021, 175, 113803, the entire contents of which are incorporated herein by reference). In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is GS-986. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is PRTX-007. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is PRX-034. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is S-34240. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is MBS-8. In some embodiments, the agonist of Toll-like receptor 7 (TLR7) is APR-002.

[0337] In some embodiments, agonists of Toll-like receptor 7 / 8 (TLR7 / 8) are selected from the group consisting of imiquimod (R837), resiquimod (R848), 852-A (PF-4878691), besifimod (GS-9620), AZD8848, motolimod (VTX-2337), selgantolimod (GS-9688), NKTR-262, RG-7854 (RO7020531), DSP-0509, BDB-001, BDC-1001, LHC-165, SHR-2150, JNJ-4964 (TQ-73334), RO-7119929, DN-1508052, VTX-1463, BNT-411 (SC1), APR-003, ALT-702, TRANSCON, VX-001, SNAPvax, R848-HA, SM360320, and GSK2245035 (see, e.g., Bhagchandani et al. Advanced Drug Delivery Reviews 2021, 175, 113803; and Evans et al. ACS Omega 2019, 4, 13, 15665, the entire contents of each of which are incorporated herein by reference). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is imiquimod (R837). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is resiquimod (R848). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is 852-A (PF-4878691). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is besifimod (GS-9620). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is AZD8848. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is motolimod (VTX-2337). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is selgantolimod (GS-9688). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is NKTR-262. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is RG-7854 (RO7020531).In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is DSP-0509. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is BDB-001. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is BDC-1001. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is LHC-165. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is SHR-2150. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is JNJ-4964 (TQ-73334). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is RO-7119929. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is DN-1508052. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is VTX-1463. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is BNT-411 (SC1). In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is APR-003. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is ALT-702. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is TRANSCON. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is VX-001. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is SNAPvax. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is R848-HA. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is SM360320. In some embodiments, the agonist of Toll-like receptor 7 / 8 (TLR7 / 8) is GSK2245035.

[0338] In some embodiments, the agonist of Toll-like receptor 8 (TLR8) is selected from the group consisting of SBT-6050, SBT-6290, and the ZM-TLR8 agonist. In some embodiments, the agonist of Toll-like receptor 8 (TLR8) is SBT-6050. In some embodiments, the agonist of Toll-like receptor 8 (TLR8) is SBT-6290. In some embodiments, the agonist of Toll-like receptor 8 (TLR8) is the ZM-TLR8 agonist.

[0339]

[0167] In certain embodiments, D is a cytotoxic agent or cytotoxic payload. In certain embodiments, D is an alkylating agent or alkylating payload. In certain embodiments, D is a bifunctional alkylating agent. In some embodiments, D is a bifunctional alkylating agent selected from the group consisting of cyclophosphamide, mechlorethamine, chlorambucil, and melphalan. In some embodiments, D is cyclophosphamide. In some embodiments, D is mechlorethamine. In some embodiments, D is chlorambucil. In some embodiments, D is melphalan. In some embodiments, D is a monofunctional alkylating agent. In some embodiments, D is a monofunctional alkylating agent selected from the group consisting of dacarbazine, nitrosourea, and temozolomide. In some embodiments, D is dacarbazine. In some embodiments, D is nitrosourea. In some embodiments, D is temozolomide. In certain embodiments, D is a cytoskeletal disrupting agent (e.g., taxane). In some embodiments, D is a cytoskeletal disrupting agent selected from the group consisting of paclitaxel, docetaxel, abraxane, and taxotere. In some embodiments, D is paclitaxel. In some embodiments, D is docetaxel. In some embodiments, D is abraxane. In some embodiments, D is taxotere. In certain embodiments, D is epothilone. In some embodiments, D is an epothilone selected from the group consisting of epothilone A, epothilone B, epothilone C, epothilone D, and ixabepilone. In some embodiments, D is epothilone A. In some embodiments, D is epothilone B. In some embodiments, D is epothilone C. In some embodiments, D is epothilone D. In some embodiments, D is ixabepilone. In certain embodiments, D is a histone deacetylase inhibitor. In some embodiments, D is a histone deacetylase inhibitor selected from the group consisting of vorinostat and romidepsin. In some embodiments, D is vorinostat. In some embodiments, D is romidepsin. In certain embodiments, D is a kinase inhibitor.In some embodiments, D is a kinase inhibitor selected from the group consisting of bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, and visomodegib. In some embodiments, D is bortezomib. In some embodiments, D is erlotinib. In some embodiments, D is gefitinib. In some embodiments, D is imatinib. In some embodiments, D is vemurafenib. In some embodiments, D is visomodegib. In certain embodiments, D is a nucleotide analog and / or a precursor analog. In some embodiments, 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). In some embodiments, D is azacitidine. In some embodiments, D is azathioprine. In some embodiments, D is capecitabine. In some embodiments, D is cytarabine. In some embodiments, D is doxifluridine. In some embodiments, D is fluorouracil. In some embodiments, D is gemcitabine. In some embodiments, D is hydroxyurea. In some embodiments, D is mercaptopurine. In some embodiments, D is methotrexate. In some embodiments, D is tioguanine (formerly thioguanine). In certain embodiments, D is a peptide antibiotic. In some embodiments, D is a peptide antibiotic selected from the group consisting of bleomycin and actinomycin. In some embodiments, D is bleomycin. In some embodiments, D is actinomycin. In certain embodiments, D is a platinum-based agent or a platinum-based payload. In some embodiments, D is a platinum-based agent or a platinum-based payload selected from the group consisting of carboplatin, cisplatin, and oxaliplatin. In some embodiments, D is carboplatin.In some embodiments, D is cisplatin. In some embodiments, D is oxaliplatin. In certain embodiments, D is a retinoid. In some embodiments, D is a retinoid selected from the group consisting of tretinoin,alitretinoin, and bexarotene. In some embodiments, D is tretinoin. In some embodiments, D is alitretinoin. In some embodiments, D is bexarotene. In certain embodiments, D is a vinca alkaloid and its derivatives. In some embodiments, D is a vinca alkaloid and its derivatives selected from the group consisting of vinblastine, vincristine, vindesine, and vinorelbine. In some embodiments, D is vinblastine. In some embodiments, D is vincristine. In some embodiments, D is vindesine. In some embodiments, D is vinorelbine. In certain embodiments, the cytotoxic agent or cytotoxic payload is a tubulin inhibitor, a DNA topoisomerase I inhibitor, or a DNA topoisomerase II inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a tubulin inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase I inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase I inhibitor selected from the group consisting of irinotecan, SN-38, topotecan, and exatecan. In some embodiments, the cytotoxic agent or cytotoxic payload is irinotecan. In some embodiments, the cytotoxic agent or cytotoxic payload is SN-38. In some embodiments, the cytotoxic agent or cytotoxic payload is topotecan. In some embodiments, the cytotoxic agent or cytotoxic payload is exatecan. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase II inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase II inhibitor selected from the group consisting of etoposide, teniposide, and tufoploside. In some embodiments, the cytotoxic agent or cytotoxic payload is etoposide.In some embodiments, the cytotoxic agent or cytotoxic payload is teniposide. In some embodiments, the cytotoxic agent or cytotoxic payload is tafuriposide. In certain embodiments, D is selected from the group consisting of hemiasterlin, camptothecin, and anthracyclines. Anthracyclines can include PNU-159682 and EDA PNU-159682 derivatives. In certain embodiments, the anthracycline is selected from the group consisting of daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, and valrubicin. In some embodiments, the anthracycline is daunorubicin. In some embodiments, the anthracycline is doxorubicin. In some embodiments, the anthracycline is epirubicin. In some embodiments, the anthracycline is idarubicin. In some embodiments, the anthracycline is mitoxantrone. In some embodiments, the anthracycline is valrubicin. In some embodiments, D is hemiasterlin. In some embodiments, D is camptothecin. In some embodiments, D is an anthracycline. In some embodiments, D is PNU-159682. In some embodiments, D is an EDA PNU compound. In some embodiments, D is an EDA PNU-159682 derivative. In certain embodiments, D is hemiasterlin, exatecan, PNU-159682, or an EDA PNU-159682 derivative. In some embodiments, D is hemiasterlin. In some embodiments, D is exatecan. In some embodiments, D is PNU-159682. In some embodiments, D is an EDA PNU-159682 compound or derivative.

[0340]

[0168] Representative conjugates of the present disclosure that include conjugates of formulas (II), (IIA), and (IIB) are shown in Table 2.

[0341]

Table 2-1

[0342]

Table 2-2

[0343]

Table 2-3

[0344]

Table 2-4

[0345]

Table 2-5

[0346]

Table 2-6

[0347]

Table 2-7

[0348]

Table 2-8

[0349]

Table 2-9

[0350]

Table 2-10

[0351]

Table 2-11

[0352]

Table 2-12

[0353]

Table 2-13

[0354]

Table 2-14

[0355]

Table 2-15

[0356]

Table 2-16

[0357]

Table 2-17

[0358]

Table 2-18

[0359]

Table 2-19

[0360]

[0169] In certain embodiments, the conjugate of formula (II), (IIA), or (IIB) is

[0361]

Chemical Structure

[0362]

Chemical Structure

[0363]

Chem.

[0364]

Chem.

[0365]

Chem.

[0366]

Chem.

[0367]

Chem.

[0368]

Chem.

[0369]

Chem.

[0370]

Chem.

[0371] [Chemical formula] is the residue of the second compound. In some embodiments, the conjugate of formula (II), (IIA), or (IIB) is

[0372] [Chemical formula] and

[0373] [Chemical formula] is the residue of the second compound. In some embodiments, the conjugate of formula (II), (IIA), or (IIB) is

[0374] [Chemical formula] and

[0375] [Chemical formula] is the residue of the second compound. In some embodiments, the conjugate of formula (II), (IIA), or (IIB) is

[0376] [Chemical formula] and

[0377] [Chemical formula] is the residue of the second compound. In some embodiments, the conjugate of formula (II), (IIA), or (IIB) is

[0378]

Chem.

[0379]

Chem.

[0380]

[0170] In certain embodiments, the conjugate of formula (II), (IIA), or (IIB) is

[0381]

Chem.

[0382]

Chem.

[0383]

Chem.

[0384]

Chem.

[0385]

Chem.

[0386] [Chemical formula] and

[0387] [Chemical formula] is a residue of the second compound.

[0388]

[0171] In certain embodiments, the conjugate of formula (II), (IIA), or (IIB) is

[0389] [Chemical formula]

[0390] [Chemical formula]

[0391] [Chemical formula]

[0392] [Chemical formula]

[0393] [Chemical formula]

[0394] [Chemical formula] selected from the group consisting of

[0395] [Chemical formula] is a residue of the second compound. In some embodiments, the conjugate of formula (II), (IIA), or (IIB) is,

[0396]

Chemical formula

[0397]

Chemical formula

[0398]

Chemical formula

[0399]

Chemical formula

[0400]

Chemical formula

[0401]

Chemical formula

[0402]

Chemical formula

[0403]

Chem.

[0404]

Chem.

[0405]

Chem.

[0406]

Chem.

[0407]

Chem.

[0408]

[0172] In certain embodiments, the conjugate of formula (II), (IIA), or (IIB) is

[0409]

Chem.

[0410]

Chem.

[0411]

Chemical Structure

[0412]

Chemical Structure

[0413]

[0173] In certain embodiments, the conjugate of formula (II), (IIA), or (IIB) is,

[0414]

Chemical Structure

[0415]

Chemical Structure

[0416]

[0174] In some aspects, the present disclosure provides a conjugate having the structure of formula (IV):

[0417]

Chemical Structure

[0418]

[0175] 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.

[0419]

[0176] In some embodiments, the compound of formula (IVA)

[0420]

Chemical formula

[0421]

[0177] In some embodiments, the compound of formula (IV) is by the compound of formula (IB)

[0422]

Chemical formula

[0423]

[0178] In some embodiments, L 1 is -C 1-3 alkylene-. In some embodiments, L 1 is -CH2-. In some embodiments, L 1 is -CH2CH2-. In some embodiments, L 1 is -CH2CH2CH2-.

[0424]

[0179] In some embodiments, Z is -X 1 -C 1-6 alkylene-[X1 -C 1-6 alkylene] n -X 1 -, -X 1 -C 2-6 alkenylene-[X 1 -C 2-6 alkenylene] n -X 1 -, -X 1 -C 2-6 alkynylene-[X 1 -C 2-6 alkynylene] n -X 1 - and at least one alkylene, alkenylene, or alkynylene of Z is substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocyclic ring, 3- to 12-membered heterocyclic ring, and C 1-10 haloalkyl. In some embodiments, Z is -X 1 -C 1-6 alkylene-[X 1 -C 1-6 alkylene] n -X 1 - and the alkylene of Z is substituted with one or more substituents selected from halogen, -CN, -NO2, -OH, -N(R 10 )2, -C(O)N(R 10 )2, -C(O)-, -C(S)-, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl, C 3-12 carbocyclic ring, 3- to 12-membered heterocyclic ring, and C 1-10 haloalkyl. In some embodiments, Z is -X 1 -C 1-6 alkylene-[X 1 -C 1-6Alkylene n -X 1 - is.

[0425]

[0180] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0426]

[0181] In some embodiments, X 1 and X 2 are independently selected from -C(O)- and -N(R 10 )C(O)-.

[0427]

[0182] In certain embodiments, RL comprises an alkyne, cyclooctyne, strained alkene, tetrazine, amine, thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide. In some embodiments, RL comprises an alkyne. In some embodiments, RL comprises cyclooctyne. In some embodiments, RL comprises a strained alkene. In some embodiments, RL comprises tetrazine. In some embodiments, RL comprises an amine. In some embodiments, RL comprises a thiol. In some embodiments, RL comprises a para-acetyl-phenylalanine residue. In some embodiments, RL comprises oxyamine. In some embodiments, RL comprises maleimide. In some embodiments, RL comprises azide. In certain embodiments, RL is

[0428]

Chemical formula

[0429]

Chemical formula

[0430]

Chem.

[0431]

Chem.

[0432]

Chem.

[0433]

Chem.

[0434]

Chem.

[0435]

Chem.

[0436]

Chem.

[0437]

Chem.

[0438]

Chem.

[0439]

Chem.

[0440]

Chem.

[0441]

Chem.

[0442]

Chem.

[0443]

Chem.

[0444]

Chem.

[0445]

Chem.

[0446]

Chem.

[0447]

Chem.

[0448]

Chem.

[0449]

Chem.

[0450]

[0183] In some embodiments, 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, Su is

[0451]

Chem.

[0452] [Chem.] represents a bond to the remainder of the compound. In certain embodiments, Su is

[0453] [Chem.] and

[0454] [Chem.] represents a bond to the remainder of the compound.

[0455]

[0184] In certain embodiments, CYTO is a cytotoxic agent or a cytotoxic payload. In certain embodiments, the cytotoxic agent or cytotoxic payload is a tubulin inhibitor, a DNA topoisomerase I inhibitor, or a DNA topoisomerase II inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a tubulin inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase I inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase I inhibitor selected from the group consisting of irinotecan, SN-38, topotecan, and exatecan. In some embodiments, the cytotoxic agent or cytotoxic payload is irinotecan. In some embodiments, the cytotoxic agent or cytotoxic payload is topotecan. In some embodiments, the cytotoxic agent or cytotoxic payload is exatecan. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase II inhibitor. In some embodiments, the cytotoxic agent or cytotoxic payload is a DNA topoisomerase II inhibitor selected from the group consisting of etoposide, teniposide, and tafuriposide. In some embodiments, the cytotoxic agent or cytotoxic payload is etoposide. In some embodiments, the cytotoxic agent or cytotoxic payload is teniposide. In some embodiments, the cytotoxic agent or cytotoxic payload is tafuriposide. In certain embodiments, CYTO is selected from the group consisting of hemiasterlin, camptothecin, and anthracyclines. Anthracyclines can include PNU-159682 and EDA PNU-159682 derivatives. In certain embodiments, the anthracycline is selected from the group consisting of daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, and valrubicin. In some embodiments, the anthracycline is daunorubicin. In some embodiments, the anthracycline is doxorubicin. In some embodiments, the anthracycline is epirubicin. In some embodiments, the anthracycline is idarubicin.In some embodiments, the anthracycline is mitoxantrone. In some embodiments, the anthracycline is valrubicin. In some embodiments, CYTO is hemiasterlin. In some embodiments, CYTO is camptothecin. In some embodiments, CYTO is an anthracycline. In some embodiments, CYTO is PNU-159682. In some embodiments, CYTO is an EDA PNU compound. In some embodiments, CYTO is an EDA PNU-159682 derivative. In certain embodiments, CYTO is hemiasterlin, exatecan, PNU-159682, or an EDA PNU-159682 derivative. In some embodiments, CYTO is hemiasterlin. In some embodiments, CYTO is exatecan. In some embodiments, CYTO is PNU-159682. In some embodiments, CYTO is an EDA PNU-159682 compound or derivative. In some embodiments, CYTO is not an immunostimulatory compound.

[0456]

[0185] In some embodiments, the conjugate is

[0457]

Chem.

[0458]

Chem.

[0459]

[0186] In some embodiments, the conjugate is

[0460]

Chem.

[0461] [Chem.] is the residue of the second compound.

[0462]

[0187] Polymer (COMP) Polymer (COMP) can be any polymer considered suitable by those skilled in the art. In certain embodiments, the polymer is a protein, peptide, antibody or its antigen-binding fragment, 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 its antigen-binding fragment. 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.

[0463]

[0188] 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 (registered trademark) (Labetuzumab), the anti-carcinoembryonic antigen (CEA) antibody developed by Immunomedics, LymphoCide (registered trademark) (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 (registered trademark) (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, Xolair® (omalizumab), the humanized anti-IgE antibody developed by Genentech and Novartis, and MLN01, the anti-β2 integrin antibody developed by Xoma are included.

[0464]

[0189] 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); Brolucizumab (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); 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 Genentics); Zanolimumab (HuMax-CD4) (CD4, Genmab); HCD122 (CD40, Novartis); SGN-40 (CD40, Seattle Genentics); 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 (Tremelimumab, 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).;

[0465]

[0190] 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-simplex 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.

[0466]

[0191] 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 include reactive groups that can conjugate with compounds or linkers of formula (I), (IA), (IB), (III), (IIIA), or (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 formula (I), (IA), (IB), (III), (IIIA), or (IIIB) linked to a residue of an antibody reactive group. In these embodiments, a precursor of a compound of formula (I), (IA), (IB), (III), (IIIA), or (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.

[0467]

[0192] Reactive group The reactive group promotes conjugation of the compounds of formula (I), (IA), (IB), (III), (IIIA), or (IIIB) described herein with a second compound such as a polymer (i.e., COMP) 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 reacts through a [3+2] alkyne-azide cycloaddition reaction, an inverse electron demand Diels-Alder ligation reaction, a thiol-electrophile reaction, or a carbonyl-oxyamine reaction as described in detail herein. In certain embodiments, the reactive group comprises an alkyne, strained alkyne, tetrazine, thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide. In certain embodiments, the reactive group is

[0468] [Chemical Formula] , -N3, or SH; R 201 is lower alkyl. In certain embodiments, R 201 is methyl, ethyl, or propyl. In some embodiments, R 201 is methyl. In some embodiments, R 201 is ethyl. In some embodiments, R 201 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.

[0469]

[0193] After conjugation, a divalent residue of the reactive group (i.e., RG’) 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.

[0470]

[0194] [3+2] alkyne-azide cycloaddition reaction

[0471]

Chemical Formula

[0472]

[0195] Advantageously, the compounds described herein that contain a terminal conjugated alkyne group (e.g., a compound according to any of formula (I), (IA), or (IB)) 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 of formula (I), (IA), or (IB) 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.

[0473]

[0196] The unique reactivity of azide and alkyne functional groups makes them useful for the selective modification of polypeptides and other biological molecules. Organic azides, especially 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).

[0474]

[0197] 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 under aqueous conditions at room temperature 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, quinine, hydroquinone, vitamin K, glutathione, cysteine, Fe 2+ , Co 2+ , and application of a potential.

[0475]

[0198] 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., RG') 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 (RG') is

[0476]

Chemical Structure

[0477]

[0199] Inverse electron demand ligation reaction

[0478]

Chemical formula

[0479]

[0200] 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, for example, at room temperature without the use of a catalyst. This reaction between tetrazine and strained alkene is commonly known to those skilled in the art as the tetrazine ligation reaction.

[0480]

[0201] 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., RG’) contains 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., RG’) is

[0481]

Chemical formula

[0482]

[0202] Thiol reaction

[0483]

Chemical formula

[0484]

[0203] Advantageously, compounds containing a terminal thiol group or a suitable electrophilic group or disulfide-forming group promote selective and efficient reactions 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 maleimide groups.

[0485]

[0204] In certain embodiments, when the conjugate is formed through a thiol-maleimide reaction, the divalent residue of the reactive group

[0486]

Chemical formula

[0487]

Chemical formula

[0488]

Chemical formula

[0489]

[0205] Carbonyl-oxyamine reaction

[0490]

Chemical formula

[0491]

[0206] Advantageously, compounds containing a terminal carbonyl or oxyamine group promote 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.

[0492]

[0207] In certain embodiments, when the conjugate 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 is formed through an oxime conjugation reaction, the divalent residue of the reactive group (e.g., RG’) is

[0493]

Chemical formula

[0494]

Chemical formula

[0495]

[0208] Other reactions

[0496] 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.

[0497]

[0210] Release reaction

[0498]

[0211] The release reaction is a reaction that acts to release the biologically active moiety of a compound or conjugate from the compounds or conjugates described herein in vivo and / or in vitro. In certain embodiments, the biologically active moiety released is a compound (e.g., a cytotoxic agent or cytotoxic payload) 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 converted to two reactive components as exemplified in these reactions, and X is a drug having nitrogen or oxygen as a heteroatom for bonding. Exemplary release reactions are shown in the following reaction scheme:

[0499]

Chemical formula

[0500]

[0212] Water-soluble polymer In certain embodiments, the conjugate comprises 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 polypeptides 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 attached 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.

[0501]

[0213] The selected polymer can be water-soluble so 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.

[0502]

[0214] 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 minimal excess unreacted protein or polymer is present) can be determined by the molecular weight of the polyethylene glycol selected and the number of available reactive groups. 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.

[0503]

[0215] The water-soluble polymer can be of any structural type including, but not limited to, linear, dendritic, 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 are described using PEG.

[0504]

[0216] 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 has the formula: X’O-(CH2CH2O) n-CH2CH2-Y’ (wherein n is an integer selected from 2 to 10,000, X’ is hydrogen or, but not limited thereto, C 1-4 is a terminal modification containing an alkyl, and Y’ is a binding point to a polypeptide) and can be represented as being linked to a polypeptide.

[0505]

[0217] 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, but are not limited to, functional groups found in the 20 common amino acids, reactive groups that react specifically with complementary functional groups present in non-naturally encoded amino acids that are inert to the 20 common amino acids (including, but not limited to, 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 azide groups and / or alkyne groups). It is noted that the other end of the PEG represented by Y’ in the above formula 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, a strong nucleophile (including, but not limited to, hydrazine, hydrazide, hydroxylamine, or semicarbazide) reacts with an aldehyde or ketone group present in a non-naturally encoded amino acid to form, where applicable, a hydrazone, oxime, or semicarbazone, 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 ketone or aldehyde groups present in a water-soluble polymer.

[0506]

[0218] Among other things, any molecular weight of PEG can be used as actually desired, including, but not limited to, about 100 Daltons (Da) to 100,000 Da or more (including, but not limited to, 0.1 to 50 kDa or 10 to 40 kDa in certain embodiments). Among other things, branched-chain PEGs can also be used that contain PEG molecules having a molecular weight (MW) in the range of 1 to 100 kDa (including, but not limited to, 1 to 50 kDa or 5 to 20 kDa) for each chain. 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.

[0507]

[0219] Generally, at least one end of the PEG molecule is available for reaction with the remainder of the compound of formula (I), (IA), or (IB). 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 hydrazide, hydrazine, hydroxylamine, or semicarbazide functional groups) 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).

[0508]

[0220] In some embodiments, a polypeptide variant having a 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.

[0509]

[0221] In certain embodiments, the water-soluble polymer is an azide-containing polymer or an acetylene-containing polymer that includes 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 are suitable for use, including poly(ethylene glycol) as well as other related polymers including poly(dextran) and poly(propylene glycol), 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 but not limited to 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.

[0510]

[0222] 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 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 part 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.

[0511]

[0223] 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).

[0512]

[0224] 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.

[0513]

[0225] In addition to these forms of PEG, the polymer can also be prepared with weak 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.

[0514]

[0226] 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 in many cases from about 6,000 Da to about 80,000 Da.

[0515]

[0227] One of ordinary skill in the art will recognize that the foregoing list of substantially water-soluble backbones is in no way exhaustive and is merely exemplary, and that all polymer materials having the qualities described herein are contemplated to be suitable for use.

[0516]

[0228] 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.

[0517]

[0229] 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.

[0518]

[0230] 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, including water and oils such as those of petroleum, animal, vegetable, or synthetic origin, e.g., 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, particularly as liquid carriers for injectable solutions. Examples of suitable pharmaceutical carriers are described in Martin, E.W., Remington’s Pharmaceutical Sciences.

[0519]

[0231] 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.

[0520]

[0232] 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 injection medium at the time of use.

[0521]

[0233] 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.

[0522]

[0234] The pharmaceutical composition may include one or more pharmaceutical excipients. Any suitable pharmaceutical excipient may be used, and those 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, glyceryl 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, minor amounts of wetting or emulsifying agents, or pH buffering agents. 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.

[0523]

[0235] In some embodiments, the pharmaceutical composition includes 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.

[0524]

[0236] 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.

[0525]

[0237] 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.

[0526]

[0238] 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.

[0527]

[0239] In some embodiments, the pharmaceutical composition comprises a surfactant. Exemplary examples of surfactants include d-α tocopherol, benzalkonium chloride, benzetonium 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.

[0528]

[0240] In some embodiments, the pharmaceutical composition comprises an anti-solidification agent. Exemplary examples of anti-solidification agents include (tertiary) calcium phosphate, hydroxymethyl cellulose, hydroxypropyl cellulose, and magnesium oxide.

[0529] 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, solubilizers, emulsifiers, gelling agents, ointment bases, penetration enhancers, preservatives, solubilizing agents, 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.

[0530]

[0242] In some embodiments, the pharmaceutical composition comprises 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.

[0531]

[0243] 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 a lipid. In some aspects, the microparticles or nanoparticles are micelles, liposomes, or polymersomes.

[0532]

[0244] In some embodiments, since water can promote the degradation of some antibodies or antigen-binding fragments thereof, anhydrous pharmaceutical compositions and dosage forms comprising conjugates are further provided herein.

[0533]

[0245] 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 manufacturing, packaging, and / or storage is anticipated, pharmaceutical compositions and dosage forms comprising lactose and at least one active ingredient comprising a primary or secondary amine can be anhydrous.

[0534]

[0246] The anhydrous pharmaceutical composition can be prepared and stored such that its anhydrous nature is maintained. Thus, the anhydrous composition can be packaged using materials known to prevent exposure to water so that they can be included in appropriate formulation 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.

[0535]

[0247] The lactose-free compositions provided herein are well known in the art and can include excipients listed, for example, 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.

[0536]

[0248] Also provided are pharmaceutical compositions and dosage forms that include one or more excipients that reduce 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.

[0537]

[0249] 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 those administrations typically bypass the subject's natural defenses against contaminants, parenteral dosage forms are typically sterile or can be sterilized before administration to the subject. Examples of parenteral dosage forms include, but are not limited to, immediately injectable solutions, dry products that can be immediately dissolved or suspended in a pharmaceutically acceptable vehicle for injection, immediately injectable suspensions, and emulsions.

[0538] <0250> 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.

[0539] <0251> Excipients that increase the solubility of one or more of the antibodies disclosed herein can also be incorporated into the parenteral dosage form.

[0540] <0252> Dosage and Unit Dosage Form In human therapeutics, the physician will determine the most appropriate pharmacology in accordance with prophylactic or therapeutic treatment, as well as in accordance with age, weight, condition, and other factors specific to the subject being treated.

[0541] <0253> 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 therapeutically effective amount of one or more prophylactic antibodies or therapeutic antibodies or antigen-binding fragments thereof.

[0542]

[0254] The amount of the conjugate or composition effective in preventing or treating a disorder or one or more of its symptoms will vary depending on the nature and severity of the disease or condition, as well as 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 or payload), 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.

[0543]

[0255] As will be readily appreciated by those skilled in the art, different therapeutic effective amounts may be applicable to different diseases and conditions. Similarly, amounts that are sufficient to prevent, manage, treat, or improve such disorders but insufficient to cause adverse effects associated with the antibodies or antigen-binding fragments thereof provided herein, or sufficient to reduce adverse effects, are 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.

[0544]

[0256] In certain embodiments, treatment or prophylaxis can be initiated with one or more loading doses of the conjugate or composition provided herein, followed by one or more maintenance doses.

[0545]

[0257] In certain embodiments, a dose of the conjugate or composition provided herein can be administered to achieve a steady-state concentration of the conjugate in the blood or serum of the subject. The steady-state concentration can be determined by measurement 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.

[0546]

[0258] 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 achieve local and systemic therapeutic effects. The intraperitoneal route can be particularly useful, for example, in the treatment of ovarian tumors.

[0547]

[0259] The conjugates provided herein can be useful in 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 cancer.

[0548]

[0260] 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.

[0549]

[0261] In some diagnostic and prognostic uses or embodiments, the conjugate can 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.

[0550]

[0262] 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.

[0551]

[0263] 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.

[0552]

[0264] In some embodiments, the kit can include the conjugate or composition provided herein, an optional second agent or composition, and instructions for providing medical personnel 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 conjugate or composition provided herein, or the second agent or composition, can include an amount that, when administered to a subject, is capable of maintaining in the subject a therapeutically effective plasma level or a prophylactically effective plasma level of the compound or composition 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.

[0553]

[0265] In some embodiments, suitable packaging is provided. As used herein, "packaging" includes a solid matrix or material that is 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.

[0554]

[0266] Preparation and Synthesis Procedures General schemes for synthesizing the compounds of formula (I), (IA), or (IB) are provided below. All other groups or variable elements are as defined in the Summary of the Invention, or in any embodiment of this specification.

[0555]

Chemical Formula

[0556]

[0267] General schemes for synthesizing the compounds of formula (III), (IIIA), or (IIIB) are provided below. All other groups or variable elements are as defined in the Summary of the Invention, or in any embodiment of this specification.

[0557]

Chemical Formula

[0558]

[0268] Conjugation The conjugate can be prepared by standard techniques. In certain embodiments, the polymer is contacted with a compound of formula (I), (IA), (IB), (III), (IIIA), or (IIIB) under conditions suitable to form a bond from the polymer to the compound of formula (I), (IA), (IB), (III), (IIIA), or (IIIB) to form the conjugate. In certain embodiments, the 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 the conjugate. In certain embodiments, the 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 the conjugate. For example, in certain embodiments, the second compound comprises tetrazine; RL comprises a strained alkene. In some embodiments, RL is

[0559]

Chemical formula

[0560]

Chemical formula

[0561]

Chemical formula

[0562]

Chemical formula

[0563]

Chemical formula

[0564]

Chemical formula

[0565]

Chemical formula

[0566]

Chemical formula

[0567]

Chemical formula

[0568] [Chemistry] It includes. 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

[0569]

[0269] The compounds provided herein can be prepared, isolated, or obtained by any method that is apparent to one of ordinary skill 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 one of ordinary skill in the art and will be known to one of ordinary skill 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 (deuterated chloroform); 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).

[0570]

[0270] For all of the following examples, standard work-up 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 are not indicative of the scope of the present disclosure.

[0571]

[0271] Unless otherwise indicated, all anhydrous solvents were obtained commercially and stored under nitrogen in Sure-Seal bottles. 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 on a Shimadzu LC equipped with a CTC IFC, using a linear gradient of B (CH3CN) in mobile phase A (water + 0.1% TFA) at a flow rate of 50 mL / min on a Phenomenex Gemini NX 5μ, C18, 110 Å, 150 × 50 mm reverse-phase column. 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.

[0572]

[0272] Example 1: Synthesis of DBCO β-glucuronide-PEG12-exatecan (107)

[0573]

Chemical formula

[0574]

[0273]

[0575]

Chemical formula

[0576]

[0274]

[0577]

Chemical formula

[0578]

[0275]

[0579]

Chemical formula

[0580]

[0276]

[0581]

Chemical formula

[0582]

[0277] Synthesis of b-Glu PNP linker fragment (8)

[0583]

[0278] Synthesis of compound (4)

[0584]

Chemical formula

[0585]

[0279] A suspension of 4-hydroxybenzaldehyde (1) (5.3 g, 43.4 mmol) and 2-chloro-N-(hydroxymethyl)acetamide (2) (5 g, 40.6 mmol) in AcOH (20 mL) was slowly added with concentrated H2SO4 (32 mL). The mixture was stirred at room temperature (22 °C) for 16 h. The resulting viscous liquid was poured into ice water (300 mL) and extracted with EtOAc (100 mL × 5). The organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude compound (3), which was dissolved in 1,4-dioxane (40 mL). Concentrated hydrochloric acid (40 mL) was added to this solution. The mixture was heated under reflux for 1 h and then concentrated in vacuo to give a residue. The residue was dissolved in dioxane:H2O (1:1, 50 mL). Et3N (9 mL) was added to this mixture, followed by Boc2O (10 g, 46 mmol). The reaction mixture was stirred at room temperature for 16 h and partitioned between EtOAc (300 mL) and water (100 mL). The organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure. The residue was purified by silica gel chromatography (220 g column, 20 - 30% EtOAc:hexane in 30 min, flow rate: 40 mL / min) to give compound (4) as an off-white solid (5.3 g). MS C 13 H 17 Calculated for C12H15NO4, 251.3; found 252.2 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 10.75 (s, 1H), 9.77 (s, 1H), 7.69 - 7.62 (m, 2H), 7.31 (t, J = 6.2 Hz, 1H), 6.96 (d, J = 8.1 Hz, 1H), 4.11 (d, J = 6.1 Hz, 2H), 1.41 (s, 9H).

[0586]

[0280] Synthesis of (7)

[0587]

Chem.

[0588]

[0281] To a solution of compound (4) (2.2 g, 8.8 mmol) and (2R,3R,4S,5S,6S)-2-bromo-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (5) (3.2 g, 8.1 mmol) in anhydrous acetonitrile (50 mL) was added Ag2O (3.7 g, 16 mmol). The suspension was stirred for 16 h under an argon atmosphere. The solid was filtered off and washed with acetonitrile (10 mL). To the combined acetonitrile solution were added i-PrOH (10 mL) and NaBH4 (300 mg, 8.1 mmol), and the mixture was stirred at room temperature. After 30 min, the reaction was quenched with water (100 mL) and extracted with EtOAc (3 × 100 mL). The organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure. The residue was purified by silica gel chromatography (220 g column, 40 - 70% EtOAc:hexane in 30 min, flow rate: 40 mL / min) to give compound (7) as a white foam (3.0 g, 5.2 mmol). MS C 26 H 35 NO 13 Calculated for, 569.2; Found 570.4 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 7.15 (q, J = 6.0 Hz, 3H), 6.98 (d, J = 8.7 Hz, 1H), 5.61 - 5.41 (m, 2H), 5.25 - 5.03 (m, 3H), 4.73 (d, J = 9.9 Hz, 1H), 4.42 (d, J = 5.3 Hz, 2H), 4.03 (tt, J = 16.4, 8.3 Hz, 2H), 3.65 (s, 3H), 2.14 - 1.92 (m, 10H), 1.41 (s, 9H), 1.33 - 1.14 (m, 2H).

[0589]

[0282] Synthesis of (8)

[0590]

Chem.

[0591] To a solution of compound (7) (2.6 g, 4.56 mmol) in anhydrous DMF (15 mL) were added DIEA (1.1 mL) and bisPNP carbonate (2 g, 6.57 mmol). The mixture was stirred at room temperature for 16 hours and directly purified by reverse-phase HPLC (Phenomenex Gemini NX 5μ, C18, 110Å, 150×50 mm, mobile phase: A: 0.1% TFA in water, B: acetonitrile, gradient: 20 - 90% B over 20 minutes, flow rate 50 mL / min). After lyophilization, compound (8) (1.8 g) was obtained as a white solid. MS C 33 H 38 N2O 17 calculated, 734.2; found 735.5 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 8.35 - 8.29 (m, 2H), 7.60 - 7.53 (m, 2H), 7.36 (dd, J = 8.4, 2.2 Hz, 1H), 7.29 (d, J = 2.3 Hz, 1H), 7.22 (t, J = 6.2 Hz, 1H), 7.09 (d, J = 8.4 Hz, 1H), 5.65 (d, J = 7.9 Hz, 1H), 5.52 (t, J = 9.6 Hz, 1H), 5.26 (s, 2H), 5.19 (dd, J = 9.8, 7.9 Hz, 1H), 5.10 (t, J = 9.7 Hz, 1H), 4.76 (d, J = 9.9 Hz, 1H), 4.03 (qd, J = 16.7, 6.2 Hz, 3H), 3.66 (s, 3H), 2.06 (s, 3H), 2.02 (d, J = 2.6 Hz, 6H), 1.39 (s, 8H), 1.30 (s, 1H).

[0592]

[0284] Synthesis of m-PEG12-DBCO-PFP (16)

[0593]

[0285] Synthesis of (11)

[0594]

Chemical Structure

[0595]

[0286] A solution of Fmoc-Dap(Boc)-COOH(9) (853 mg, 2 mmol) in anhydrous DMF (10 mL) was added with N,N,N’,N’-tetramethyl-O-(N-succinimidyl)uronium tetrafluoroborate (TSTU) (608 mg, 2 mmol), followed by DIPEA (0.7 mL). The mixture was stirred at room temperature for 10 minutes. A solution of β-alanine (0.2 g) in acetonitrile:water (1:1, 3 mL) was added, followed by DIPEA (0.4 mL). The reaction mixture was stirred at room temperature for 30 minutes and then acidified with 0.5 N hydrochloric acid (50 mL). The mixture was extracted with EtOAc (200 mL), the organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure to obtain the crude compound (10) as a white powder. The crude compound (10) was treated with TFA:DCM (1:4, v / v, 20 mL) at room temperature for 1 hour. The mixture was evaporated to dryness under reduced pressure to obtain the crude compound (11), which was used directly in the next step. MS C 21 H 23 Calculated for C19H24N3O5, 397.16; found 398.2 [M+H] + 。

[0596]

[0287] Synthesis of (14)

[0597]

Chem.

[0598]

[0288] m-PEG12-acid (12) (1.18 g, 2 mmol) was dissolved in DMF (10 mL), TSTU (610 mg, 2 mmol) was added, followed by DIPEA (700 μL). After 5 minutes, a solution of compound (11) in DMF (10 mL) and DIPEA (0.7 mL) was added. The reaction mixture was stirred at room temperature for 30 minutes and directly purified by reverse-phase HPLC to obtain compound (14) as a viscous foam (1.27 g). MS C 47 H 73 N3O 18 Calculated for C47H77N3O14, 967.5; found 968.8 [M+H] + ; 11H NMR (500 MHz, DMSO-d6) δ 8.35 - 8.29 (m, 2H), 7.60 - 7.53 (m, 2H), 7.36 (dd, J = 8.4, 2.2 Hz, 1H), 7.29 (d, J = 2.3 Hz, 1H), 7.22 (t, J = 6.2 Hz, 1H), 7.09 (d, J = 8.4 Hz, 1H), 5.65 (d, J = 7.9 Hz, 1H), 5.52 (t, J = 9.6 Hz, 1H), 5.26 (s, 2H), 5.19 (dd, J = 9.8, 7.9 Hz, 1H), 5.10 (t, J = 9.7 Hz, 1H), 4.76 (d, J = 9.9 Hz, 1H), 4.03 (qd, J = 16.7, 6.2 Hz, 3H), 3.66 (s, 3H), 2.06 (s, 3H), 2.02 (d, J = 2.6 Hz, 6H), 1.39 (s, 8H), 1.30 (s, 1H).

[0599]

[0289] Synthesis of (15)

[0600]

Chem.

[0601]

[0290] To a solution of compound (14) (1.24 g) in DMF (10 mL) was added i-Pr2NH (DIPA) (10 mL), and the reaction mixture was stirred at room temperature for 2 hours. Then, the mixture was concentrated under reduced pressure to about 9 mL. To this solution was added DBCO-C6-NHS (0.55 g), followed by the addition of DIPEA (0.23 mL). The mixture was stirred at room temperature for 2 hours and then purified directly by reverse-phase HPLC to obtain compound (15) as a colorless syrup (1.17 g). MS C 53 H 80 N4O 18 Calculated value for, 1060.6; Found 1061.9 [M + H] + .

[0602]

[0291] Synthesis of m-PEG12-DBCO-PFP linker (16)

[0603]

Chem.

[0604]

[0292] To a solution of compound (15) (1.1 g) in DMF (8 mL) was added pentafluorophenol-tetramethyluronium hexafluorophosphate (PfTU) (0.5 g), followed by DIEA (0.4 mL), and the reaction mixture was stirred at room temperature for 10 minutes. Direct purification of the mixture by RP-HPLC gave compound (16) as a colorless syrup (1.0 g). MS C 59 H 79 F5N4O 18 Calculated value of, 1226.5; found 1227.8 [M+H] + . Compound (16) was used immediately in the next reaction.

[0605]

[0293] Synthesis of (107)

[0606]

[0294] Synthesis of (19)

[0607]

Chemical formula

[0608] To a solution of compound (8) (735 mg, 1 mmol) and exatecan mesylate (531 mg, 1 mmol) in DMF (10 mL) was added DIPEA (350 μL). The reaction mixture was stirred at room temperature (22 °C) for 5 h and then diluted with EtOAc (200 mL). The mixture was washed with 0.5 N hydrochloric acid (100 mL), water (100 mL), and brine (50 mL). The organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure to give the crude compound (18), which was suspended in acetonitrile:water (2:1, 50 mL). To this mixture was added 1 N aqueous NaOH (7 mL), and the reaction was stirred at room temperature. After 3 h, 1 N hydrochloric acid (7 mL) was added, and the mixture was evaporated to dryness under reduced pressure. The resulting residue was treated with TFA:DCM (1:4, v / v, 20 mL) at room temperature for 1 h. The mixture was diluted with toluene (30 mL) and then evaporated to dryness under reduced pressure. Purification of the residue by reverse-phase HPLC gave compound (19) as a yellow solid (745 mg). MS C 39 H 39 FN4O 13 calculated for, 790.3; found 791.6 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 8.09 (d, J = 8.0 Hz, 4H), 7.73 (dd, J = 10.7, 3.2 Hz, 1H), 7.54 (d, J = 2.1 Hz, 1H), 7.49 (dd, J = 8.5, 2.2 Hz, 1H), 7.32 (s, 1H), 7.22 (d, J = 8.5 Hz, 1H), 6.55 (s, 1H), 5.75 (s, 1H), 5.48 (d, J = 16.3 Hz, 1H), 5.46 - 5.37 (m, 3H), 5.31 - 5.13 (m, 4H), 5.08 - 5.02 (m, 2H), 4.19 - 4.07 (m, 2H), 3.95 (d, J = 9.5 Hz, 1H), 3.44 (dd, J = 8.8, 2.6 Hz, 1H), 3.28 - 3.19 (m, 2H), 3.14 - 3.04 (m, 1H), 2.34 (s, 3H), 2.26 (dd, J = 12.5, 6.4 Hz, 1H), 2.12 (qd, J = 9.0, 4.6 Hz, 1H), 1.87 (dh, J = 21.5, 7.2 Hz, 2H), 0.89 (t, J = 7.2 Hz, 3H).

[0609] Synthesis of [

[0296] ] (107)

[0610] [Chemical formula]

[0611]

[0297] Compound b-Glu-exatecan benzylamine (19) (540 mg, 0.68 mmol) was dissolved in DMF (4 mL), and compound m-PEG12-DBCO-PFP (16) (750 mg, 0.6 mmol) and then DIPEA (0.32 mL) were added. The reaction mixture was stirred at room temperature for 30 minutes and purified directly by reverse-phase HPLC (Phenomenex Gemini NX 5μ, C18, 110Å, 150×50 mm, mobile phase: A: 0.1% TFA in water, B: acetonitrile, gradient: 20 - 90% B over 20 minutes, flow rate 50 mL / min), and compound (107) was obtained as a pale yellow solid (704 mg). HRMS m / z (ESI + ): C 92 H 117 FN8O 30 Calculated value for, 1832.78; Measured value 1833.79 [M+H] + ; 11H NMR (500 MHz, DMSO-d6) δ 12.84 (s, 1H), 8.20 (t, J = 6.2 Hz, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.75 (td, J = 6.1, 3.4 Hz, 2H), 7.62 - 7.52 (m, 1H), 7.50 - 7.39 (m, 2H), 7.32 - 7.25 (m, 2H), 6.52 (s, 1H), 5.50 - 5.39 (m, 2H), 5.27 (d, J = 13.6 Hz, 3H), 5.01 - 4.94 (m, 1H), 4.29 (t, J = 6.3 Hz, 2H), 3.57 (d, J = 14.0 Hz, 1H), 3.56 - 3.46 (m, 35H), 3.48 - 3.40 (m, 5H), 3.35 (td, J = 12.8, 6.5 Hz, 5H), 3.24 (s, 2H), 3.22 (s, 3H), 2.36 (d, J = 1.8 Hz, 3H), 2.27 (dq, J = 13.4, 6.3 Hz, 3H), 2.16 (ddd, J = 21.5, 11.0, 6.0 Hz, 2H), 1.87 (dp, J = 21.0, 7.2 Hz, 4H), 1.17 (s, 3H), 0.88 (t, J = 7.3 Hz, 3H).

[0612]

[0298] Synthesis of β-glucuronide-cleavable hemiasterlin (114)

[0613]

Chem.

[0614]

[0299]

[0615]

Chem.

[0616]

[0300] Synthesis of (5)

[0617]

Chem.

[0618]

[0301] A solution of compound (1) DBCO-C6-NHS ester (150 mg, 0.35 mmol), compound (2) β-alanine (32 mg, 0.70 mmol), and DIEA (112 mg, 0.87 mmol) in 50% CH3CN:H2O (5 mL) was stirred at room temperature for 20 minutes. The solution was extracted with dichloromethane (50 mL), and the organic layer was concentrated to obtain crude compound (3), which was redissolved in anhydrous DCM (5 mL). To this solution, N-hydroxysuccinimide (129 mg, 0.70 mmol) and EDC·HCl (200 mg, 1.05 mmol) were sequentially added, and the reaction mixture was stirred at room temperature for 2 hours. The solvent was removed under vacuum. The residue was dissolved in DMF (3 mL) and purified by reverse-phase HPLC to obtain compound (5) as an amorphous solid (136 mg).

[0619]

[0302]

[0620]

Chem.

[0621]

[0303]

[0622]

Chem.

[0623]

[0304] Synthesis of (114)

[0624]

Chem.

[0625] To a solution of compound (6) (100 mg, 136 μmol) and compound (7) as the TFA salt (100 mg, 134 μmol) in DMF (2 mL) was added DIPEA (75 μL). The reaction mixture was stirred at room temperature (22 °C) for 3 days, and the crude product was purified by reverse-phase HPLC. After lyophilization, compound (8) was obtained as a white solid (143 mg, 86%). Compound (8) (143 mg, 117 μmol) was treated with TFA:DCM (1:4, v / v, 3 mL) at room temperature for 30 minutes. The reaction was concentrated to dryness under reduced pressure to give a residue. The residue was dissolved in acetonitrile:water (6:4, v / v, 3 mL), and an aqueous NaOH solution (1 M, 0.8 mL) was added. The reaction mixture was stirred at room temperature for 2 days and acidified with hydrochloric acid (1 M, 0.5 mL). The mixture was purified by reverse-phase preparative HPLC. After lyophilization, compound (9) (87 mg, 69% over two steps) was obtained as a white solid. Compound (9) (87 mg) was dissolved in DMF (2 mL), compound (5) (41 mg, 82 μmol) was added, followed by DIPEA (70 μL). The reaction mixture was stirred at room temperature for 20 minutes and directly purified by reverse-phase preparative HPLC (Phenomenex Gemini NX 5μ, C18, 110 Å, 150×50 mm, mobile phase: A: 0.1% TFA in water, B: acetonitrile, gradient: 5 - 60% B over 30 minutes, flow rate 50 mL / min), and compound (144) was obtained as a white solid (87 mg, 84%). MS C 66 H 83 N7O 16 Calculated for, 1229.59; found m / z 1230.7 [M + H] + 。 11H NMR (500 MHz, DMSO-d6) δ 9.65 (s, 1H), 8.70 (s, 1H), 7.99 (s, 1H), 7.66 (t, J = 5.6 Hz, 1H), 7.61 (dd, J = 7.6, 1.5 Hz, 1H), 7.57 - 7.40 (m, 5H), 7.40 - 7.24 (m, 6H), 7.20 (t, J = 7.9 Hz, 1H), 7.09 (dd, J = 13.4, 8.1 Hz, 2H), 6.64 (dd, J = 9.6, 1.7 Hz, 1H), 5.48 (s, 1H), 5.17 (s, 1H), 5.07 - 4.99 (m, 3H), 4.91 (t, J = 10.1 Hz, 1H), 4.76 (d, J = 8.8 Hz, 2H), 4.37 (dd, J = 14.7, 6.5 Hz, 1H), 4.23 (dd, J = 14.8, 5.6 Hz, 1H), 3.59 (d, J = 13.8 Hz, 2H), 3.18 (qd, J = 6.8, 2.6 Hz, 3H), 2.98 (s, 3H), 2.27 (t, J = 7.2 Hz, 2H), 2.13 (ddd, J = 13.9, 8.0, 5.6 Hz, 1H), 2.06 - 1.93 (m, 4H), 1.83 - 1.75 (m, 5H), 1.71 (dtd, J = 14.5, 5.9, 2.7 Hz, 1H), 1.26 (s, 4H), 1.16 (d, J = 20.8 Hz, 6H), 0.93 (s, 9H), 0.79 (d, J = 6.6 Hz, 3H), 0.73 (d, J = 6.5 Hz, 3H).

[0626]

[0306] Synthesis of (108)

[0627]

Chem.

[0628]

[0307]

[0629]

Chem.

[0630] Compound (9) (60 mg, 55 μmol) was dissolved in anhydrous DMF (2 mL), compound (16) (74 mg, 60 μmol) was added, and subsequently DIPEA (60 μL) was added. The reaction mixture was stirred at room temperature for 20 minutes and directly purified by reverse phase preparative HPLC (Phenomenex Gemini NX 5 μ, C18, 110 Å, 150×50 mm, mobile phase: A: 0.1% TFA in water, B: acetonitrile, gradient: 5 - 60% B over 30 minutes, flow rate 50 mL / min), and compound (108) was obtained as a white solid (93 mg, 84%). HRMS m / z (ESI + ): C 95 H 139 N9O 30 calculated for, 1885.96; found 1886.97 [M+H] + ; 11H NMR (500 MHz, DMSO-d6) δ 12.61 (s, 2H), 9.77 (s, 1H), 8.76 (d, J = 8.2 Hz, 2H), 8.20 (t, J = 6.1 Hz, 1H), 7.85 (t, J = 5.8 Hz, 1H), 7.76 (t, J = 5.9 Hz, 2H), 7.67 (dd, J = 8.0, 2.7 Hz, 1H), 7.64 - 7.52 (m, 3H), 7.53 - 7.41 (m, 3H), 7.41 - 7.25 (m, 6H), 7.25 - 7.15 (m, 2H), 7.07 (d, J = 8.5 Hz, 1H), 6.67 (dd, J = 9.6, 1.7 Hz, 1H), 5.57 (s, 2H), 5.17 - 4.99 (m, 3H), 5.00 - 4.87 (m, 2H), 4.77 (d, J = 8.1 Hz, 1H), 4.30 (ddt, J = 31.2, 20.7, 7.9 Hz, 3H), 4.22 - 4.10 (m, 1H), 3.91 (d, J = 9.7 Hz, 1H), 3.69 - 3.37 (m, 70H), 3.37 - 3.28 (m, 5H), 3.23 (s, 9H), 3.02 (s, 3H), 2.28 (q, J = 5.9 Hz, 7H), 2.17 (ddd, J = 16.6, 9.0, 3.8 Hz, 1H), 2.01 (ddd, J = 10.5, 7.9, 5.0 Hz, 1H), 1.95 - 1.85 (m, 2H), 1.79 (d, J = 1.4 Hz, 3H), 1.74 (ddd, J = 14.2, 7.0, 2.8 Hz, 1H), 1.33 (s, 4H), 1.26 - 1.10 (m, 6H), 0.98 (s, 10H), 0.80 (d, J = 6.5 Hz, 3H), 0.77 (d, J = 6.5 Hz, 3H).

[0631]

[0309] Synthesis of (109)

[0632]

Chem.

[0633]

[0310]

[0634]

Chem.

[0635]

[0311] To a solution of compound (1) (50 mg) in DCM (2 mL), pentafluorophenol (20 mg) was added, followed by DIC (8 mL), and the reaction mixture was stirred at room temperature for 5 minutes under an argon atmosphere. To this solution, a solution of compound (19) (45 mg) in DMF (2 mL) was added, followed by DIPEA (0.04 mL). The reaction mixture was stirred at room temperature for 1 hour. LCMS indicated the completion of the reaction. DCM was removed under reduced pressure, and the residue was purified directly by reverse-phase HPLC to obtain compound (109) as a pale yellow solid (44 mg). LCMS m / z (ESI + ): C 89 H 112 FN7O 29 calculated value for, 1761.75; measured value 1762.8 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 12.80 (s, 1H), 8.29~8.14 (m, 1H), 8.00 (d, J = 8.8 Hz, 1H), 7.78 (tt, J = 14.3, 6.8 Hz, 3H), 7.58 (ddd, J = 7.4, 4.5, 1.7 Hz, 1H), 7.53 (dd, J = 7.0, 1.8 Hz, 1H), 7.44 (ttd, J = 7.6, 5.0, 2.7 Hz, 3H), 7.38~7.22 (m, 5H), 7.22~7.14 (m, 1H), 7.07 (d, J = 8.2 Hz, 1H), 6.53 (d, J = 5.0 Hz, 1H), 5.57~5.32 (m, 4H), 5.27 (s, 4H), 5.13~4.86 (m, 4H), 4.44~4.16 (m, 3H), 3.90 (d, J = 9.7 Hz, 1H), 3.60~3.52 (m, 2H), 3.53~3.37 (m, 42H), 3.38 (s, 51H), 3.24 (s, 5H), 2.38 (d, J = 1.9 Hz, 3H), 2.29~2.02 (m, 5H), 2.00~1.77 (m, 4H), 1.77~1.62 (m, 1H), 1.23 (dd, J = 57.4, 11.9 Hz, 4H), 0.94~0.75 (m, 3H).

[0636]

[0312] Synthesis of (110)

[0637]

Chem.

[0638]

[0313]

[0639]

Chem.

[0640]

[0314] To a solution of compound (1) (88 mg) in DMF (2 mL) were added PfTU (35 mg) and subsequently DIPEA (0.03 mL), and the reaction mixture was stirred at room temperature for 10 minutes. Direct purification of the mixture by reverse-phase HPLC gave compound (2) as a colorless syrup (85 mg).

[0641]

[0315] Compound (9) (45 mg) was dissolved in DMF (2 mL), compound (2) (64 mg, 0.05 mmol) was added, and subsequently DIPEA (0.04 mL) was added. The reaction mixture was stirred at room temperature for 30 minutes and direct purification by reverse-phase HPLC gave compound (110) as a pale yellow solid (57 mg). LCMS m / z (ESI + ): C 95 H 123 FN8O 30 calculated value, 1874.83; found 1875.9 [M + H] + ; 11H NMR (500 MHz, DMSO-d6) δ 12.82 (s, 2H), 8.18 (t, J = 6.0 Hz, 1H), 8.03 (d, J = 8.8 Hz, 1H), 7.84 (t, J = 5.8 Hz, 1H), 7.76 (dt, J = 11.3, 4.1 Hz, 2H), 7.70 (d, J = 8.0 Hz, 1H), 7.60 (dd, J = 7.5, 1.5 Hz, 1H), 7.55 (dt, J = 6.8, 1.9 Hz, 1H), 7.51 - 7.39 (m, 3H), 7.39 - 7.18 (m, 6H), 7.09 (d, J = 8.5 Hz, 1H), 6.52 (s, 1H), 5.55 (s, 1H), 5.50 - 5.32 (m, 3H), 5.27 (s, 4H), 5.13 - 4.86 (m, 4H), 4.40 - 4.19 (m, 2H), 4.06 (tdd, J = 8.3, 5.1, 3.0 Hz, 1H), 3.91 (d, J = 9.7 Hz, 1H), 3.17 - 3.01 (m, 1H), 2.96 (dd, J = 10.4, 4.5 Hz, 2H), 2.37 (d, J = 1.9 Hz, 3H), 2.27 (q, J = 6.6 Hz, 4H), 2.16 (ddd, J = 21.5, 10.9, 5.8 Hz, 3H), 1.88 (tp, J = 14.0, 6.2 Hz, 4H), 1.79 - 1.64 (m, 1H), 1.51 (s, 1H), 1.38 (d, J = 9.3 Hz, 1H), 1.29 (d, J = 8.9 Hz, 3H), 1.16 (dq, J = 15.0, 7.3 Hz, 4H), 0.88 (t, J = 7.4 Hz, 3H).

[0642]

[0316] Synthesis of (111)

[0643]

Chem.

[0644]

[0317]

[0645]

Chem.

[0646]

[0318] (111) was synthesized in the same manner using the same method as above. LCMS m / z (ESI + ): C95 H 123 FN8O 30 Calculated value, 1804.75; measured value 1804.90 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 12.82 (s, 1H), 8.39 - 8.10 (m, 1H), 8.11 - 7.93 (m, 1H), 7.93 - 7.54 (m, 6H), 7.54 - 7.36 (m, 3H), 7.40 - 7.16 (m, 6H), 7.10 (dd, J = 8.6, 4.3 Hz, 1H), 6.53 (d, J = 5.2 Hz, 2H), 5.61 - 5.30 (m, 4H), 5.26 (d, J = 9.2 Hz, 4H), 5.15 - 4.69 (m, 4H), 4.42 - 4.22 (m, 2H), 4.22 - 4.07 (m, 1H), 3.91 (d, J = 9.5 Hz, 1H), 3.66 - 3.53 (m, 3H), 3.54 - 3.45 (m, 36H), 3.45 - 3.39 (m, 6H), 3.39 - 3.28 (m, 63H), 3.24 (s, 5H), 3.16 - 2.96 (m, 2H), 2.60 (dt, J = 15.8, 7.2 Hz, 1H), 2.41 - 2.27 (m, 5H), 2.23 (t, J = 6.7 Hz, 3H), 2.21 - 2.11 (m, 2H), 2.03 (dq, J = 14.3, 7.4 Hz, 1H), 1.94 - 1.71 (m, 3H), 0.88 (t, J = 7.3 Hz, 3H).

[0647]

[0319] Synthesis of maleimidocaproyl PEGylated β-glucuronide cleavable linker payload (111a)

[0648]

Chem.

[0649]

[0320] Preparation of (15a)

[0650]

[0321] To a solution of compound (14a) (0.2 g) in DMF (2 mL) was added i-Pr2NH (2 mL), and the reaction mixture was stirred at room temperature for 2 hours. Then, the mixture was concentrated under reduced pressure to about 2 mL, and to this solution was added MC-Pfp (76 mg, CAS 692739-25-6), followed by the addition of DIEA (0.035 mL). The mixture was stirred at room temperature for 2 hours, and the crude material was purified directly by reverse-phase preparative HPLC to give compound (15a) as a colorless syrup (0.14 g). LCMS m / z (ESI + ): C 42 H 74 N4O 19 Calculated for, 938.49; found 939.63 [M+H] + .

[0651]

[0322] Preparation of (16a)

[0652]

[0323] To a solution of compound (15a) (0.14 g) in DMF (2 mL) were added PfTU (TCI-US, 0.065 g), followed by DIEA (0.06 mL), and the reaction mixture was stirred at room temperature for 10 minutes. LCMS indicated the formation of the desired product, and the crude mixture was purified directly by reverse-phase preparative HPLC to give compound (16a) as a colorless syrup (0.12 g). LCMS m / z (ESI + ): C 48 H 73 F5N4O 19 Calculated for, 1104.48; found 1105.6 [M+H] + .

[0653]

[0324] Preparation of (111a)

[0654]

[0325] Compound (9a) (92 mg) was dissolved in DMF (2 mL), compound (16a) (110 mg, 0.1 mmol) was added, followed by DIPEA (0.07 mL). The reaction mixture was stirred at room temperature for 30 minutes, and then LCMS indicated the formation of the desired product. The crude material was purified directly by reverse-phase preparative HPLC to give compound 111a as a pale yellow solid (120 mg). 11H NMR (500 MHz, DMSO-d6) δ 12.82 (s, 2H), 8.21 (t, J = 6.1 Hz, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.89 (t, J = 5.7 Hz, 1H), 7.83 - 7.69 (m, 3H), 7.31 (d, J = 7.0 Hz, 2H), 7.25 (d, J = 2.2 Hz, 1H), 7.10 (d, J = 8.4 Hz, 1H), 6.99 (s, 2H), 6.51 (s, 1H), 5.45 (d, J = 4.1 Hz, 3H), 5.28 (s, 4H), 5.07 (d, J = 6.1 Hz, 3H), 4.97 (d, J = 6.9 Hz, 1H), 4.45 - 4.12 (m, 4H), 3.92 (d, J = 9.7 Hz, 2H), 3.56 (s, 2H), 3.54 - 3.40 (m, 45H), 3.40 - 3.27 (m, 13H), 3.24 (s, 8H), 3.16 - 3.04 (m, 2H), 2.38 (d, J = 1.9 Hz, 3H), 2.31 (q, J = 5.5 Hz, 4H), 2.07 (d, J = 7.4 Hz, 2H), 1.88 (dd, J = 13.1, 7.1 Hz, 2H), 1.51 - 1.41 (m, 4H), 1.18 (s, 2H), 0.89 (t, J = 7.3 Hz, 3H); 13 13C NMR (126 MHz, DMSO-d6) δ 172.92, 172.54, 171.53, 171.08, 170.98, 170.52, 170.23, 163.11, 161.13, 157.23, 156.53, 155.10, 153.01, 150.49, 148.51, 148.40, 145.69, 141.39, 136.94, 134.90, 130.91, 128.91, 128.76, 125.89, 124.20, 124.04, 122.05, 119.60, 115.55, 110.20, 101.57, 97.18, 76.10, 75.94, 73.57, 72.84, 71.84, 71.76, 70.26, 70.24, 70.12, 70.06, 69.95, 67.19, 66.16, 65.76, 58.52, 53.26, 50.28, 47.77, 36.49, 35.93, 35.59, 35.52, 31.16, 30.80, 28.24, 26.30, 24.99, 24.21, 11.46, 11.42, 8.22; LCMS m / z (ESI + ): C 81 H 111 FN8O31 Calculated value: 1710.73; measured value: 1711.9 [M+H] + 。

[0655]

[0326] Synthesis of mDPR PEGylated β-glucuronide cleavable linker payload (111b)

[0656]

Chem.

[0657]

[0327] (111b) was synthesized in the same manner as the above (111a) (for example, instead of (15a),

[0658]

Chem.

[0659]

[0328] Synthesis of β-Glucuronide-Cleavable EDA PNU-159682 (112)

[0660]

Chem.

[0661]

[0329]

[0662]

Chem.

[0663]

[0330] To a solution of compound (8) (147 mg, 0.2 mmol) in anhydrous DMF (2 mL) were added mono-Fmoc-ethylenediamine (1) (HCl salt, 64 mg, 0.2 mmol) and subsequently DIPEA (0.07 mL). The mixture was stirred at room temperature for 1 hour and partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was dried over Na2SO4 and concentrated to dryness under reduced pressure to give crude compound (2). The crude compound (2) was treated with TFA:DCM (1:4, v / v, 4 mL) at room temperature for 30 minutes. The mixture was evaporated to dryness under reduced pressure to give crude compound 3, which was used directly in the next step.

[0664]

[0331] The crude compound (3) was dissolved in DMF (2 mL), DBCO-PEG12-Pfp ester (240 mg, 0.2 mmol) was added, and subsequently DIPEA (70 μL) was added. The reaction mixture was stirred at room temperature for 2 hours and purified directly by reverse-phase HPLC to give compound (4) as a viscous syrup (240 mg).

[0665]

[0332] To a solution of compound 4 (240 mg) in ACN:water (3:2, v / v, 6 mL) was added NaOH (aqueous solution, 1 N, 1.2 mL), and the reaction mixture was stirred at room temperature for 6 hours. Then the mixture was acidified with 1 N hydrochloric acid (0.6 mL) and purified directly by reverse-phase HPLC to give compound (5) as a colorless syrup (138 mg).

[0666]

[0333] To a solution of compound (6) (36.4 mg, 50 μmol) in anhydrous DMF (1 mL) were added TSTU (15 mg, 50 μmol) and DIPEA (18 μL, 0.1 mmol). The mixture was stirred at room temperature for 10 minutes, and then a solution of compound (5) (78.6 mg, 50 μmol) in anhydrous DMF (1 mL) was added. The reaction mixture was stirred for an additional 10 minutes and then purified directly by reverse-phase HPLC. Freeze-drying the fractions gave compound (112) as a red solid (42 mg). LCMS m / z (ESI + ): C 101 H 134 N8O 38 Calculated for: 2066.88; found 2067.9 [M+H] + .

[0667]

[0334] In certain embodiments, linker-payload compounds and / or their conjugates that contain or have an immunomodulatory payload are also contemplated herein. Linker-immunomodulatory payloads and / or their conjugates can be synthesized according to standard techniques known in the art, as will be recognized by those of skill in the art, and / or via the methods described herein for making linker-payloads and / or their conjugates (i.e., linker-payloads and / or their conjugates that contain or have a cytotoxic agent or cytotoxic payload, or are synthesized via the methods described herein for making linker-cytotoxic agents or linker-cytotoxic payloads).

[0668]

[0335] Conjugation method: (114), (107), and (108) linker-payloads were 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 as shown in Figure 1. 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.

[0669]

[0336] Results: As shown in Figure 1, (114), (107), and (108) were conjugated to the aFolR mAb by incorporating eight pAMF sites into the heavy chain Y180F404 site and the light chain K42E161 site. Conjugation efficiencies of over 94% were achieved for all three different linker-payloads according to the conjugation conditions described in the Methods section. The results via analytical SEC showed that all conjugates exhibited high purity with over 99% monomer.

[0670]

Table 3

[0671]

[0337] In vitro cell killing activity of β-glucuronide linker payload antibody-drug conjugate (ADC)

[0672]

[0338] To evaluate the cell killing activity of ADCs with different β-Glu linkers to payloads, an anti-FolRα antibody was conjugated to (114), (107), and (108) at a DAR = 8 and tested in an in vitro cell killing assay. An anti-FolRα antibody conjugated to hemiasterlin using a cathepsin-cleavable ValCit linker was used as a control. FolRα-negative A549 and MC38 cells were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA), and FolRα-positive Igrov1 cells were licensed from NCI (National Cancer Institute at Frederick, Maryland). MC38-hFolRα cells were generated at Sutro by stable transfection of a vector expressing human FolRα. 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 ADCs was measured in a cell proliferation assay. A total of 625 cells for Igrov1 and A549 and 312 cells for MC38-hFolRα in a volume of 25 microliters were seeded into 384-well flat-bottom white polystyrene plates the day before the actual assay start. The ADCs were formulated in cell culture medium at a 2× starting concentration and filtered through a MultiScreen HTS 96-well filter plate (Millipore, Billerica, MA). The filter-sterilized samples were 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 Igov1 and A549 and 72 hours for MC38-hFolRα cells. For cell viability measurement, 30 microliters 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 fit with a logarithmic (inhibitor) vs. response, variable slope nonlinear regression analysis and a four-parameter fit equation using GraphPad Prism.

[0673]

[0339] As free payloads, exatecan and hemiasterlin showed good cell killing against all three cell lines tested. As shown in Table 1A and FIGS. 2D - 2F, free exatecan (207) was approximately three-fold more potent than free hemiasterlin (201). As expected, anti-FolRα ADCs with b-Glu or b-Glu-PEG12 linkers showed potent cell killing against FolRα-positive Igrov1 cells and MC38-hFolRα cells, but no cell killing was observed in hFolRα-negative A549 cells. This indicates that there is no non-specific release of free payloads that kill target-negative cells, meaning that the b-Glu linker and the b-Glu-PEG12 linker were stable in cell culture medium for 5 days.

[0674]

[0340] As shown in Table 1A and FIGS. 2A - 2C, ADCs conjugated with hemiasterlin by β-Glu (202) or β-Glu-PEG12 linker (203) showed good similar killing activity, which was slightly weaker than that of hemiasterlin ADC (204) containing a ValCit linker. Exatecan ADC (205) containing a b-Glu-PEG12 linker was much weaker against Igov1 cells than hemiasterlin ADC (203) containing the same linker and was inactive in both MC38-hFolRα cells.

[0675]

Table 4

[0676]

[0341] β-Glucuronidase Reactivity

[0677]

[0342] The sensitivity of the β-glucuronide linker-payload to enzymatic cleavage was evaluated by treating the (107) and (108) linker-payloads with β-glucuronidase as shown in FIGS. 3A-3D. This assay enabled confirmation of the expected release of exatecan (207) and 3-aminohemicastrine (201). LCMS was used to monitor the cleavage products. 945 μL of 50 mM sodium acetate, 5 μL of 2 mM (107) / (108) linker-payload stock (final concentration 10 μM), and 50 μL of 6 KU / mL E. coli β-glucuronidase in 0.2% NaCl (final concentration 300 U / mL) were mixed and incubated at 37° C. Aliquots (50 μL) were taken at t = 0 h, 0.5 h, 1 h, 2 h, 4 h, and 24 h, treated with three volumes of quenching solution (methanol:acetonitrile 5:95 v / v), and analyzed by LCMS. After 30 minutes, most of the (107) and (108) in the reaction mixture disappeared simultaneously with the appearance of (207) and (201), and the remaining (107) and (108) were cleaved within 24 hours.

[0678]

[0343] β-Glucuronidase Cleavage of (107)

[0679]

Chem.

[0680]

[0344] Synthesis of DBCO-Valcit-pAB-Hemicastrine (208):

[0681]

Chem.

[0682]

[0345] 208 DBCO-Valcit-pAB-Hemicastrine Linker-Payload was synthesized as described in International Publication No. WO 2020 / 252015.

[0683]

[0346] β-glucuronidase cleavage of (108)

[0684]

Chem.

[0685] equivalent

[0347] The above disclosure may include 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 exemplified 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 considered 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 are also considered to be included within the subject matter of the present disclosure, whether they relate to different embodiments or the same embodiment, and whether they are broader, narrower, equal, or different in scope compared to the original claims.

[0686]

[0348] One or more features of any embodiment described herein or in the drawings may be combined with one or more features of any other embodiment described herein or in the drawings without departing from the scope of the present disclosure.

[0687]

[0349] All publications, patents, and patent applications cited in this specification are hereby incorporated by reference into this specification 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 skilled in the art that certain changes and modifications may be made without departing from the spirit or scope of the appended claims in light of the teachings of this disclosure.

Claims

1. Compounds based on the structure of formula (I) 【Chemistry 1】 or a pharmaceutically acceptable salt thereof (in the formula, L １ Ha-C １－６ Alkylene- is; Y is -X １ -C １－６ alkylene-[X １ -C １－６ alkylene] ｎ -[X １ ｐ -,-X １ -C ２－６ alkenylene-[X １ -C ２－６ alkenylene] ｎ -[X １ ｐ -、or -X １ -C ２－６ alkynylene-[X １ -C ２－６ alkynylene] ｎ -[X １ ｐ - and at least one alkylene, alkenylene, or alkynylene of Y is substituted with one or more substituents selected from R ５０ ​​​​ Alkilen, alkenylene, or alkynylene of Y is R ５１ They may be substituted with one or more substituents selected from; R ５０ is, -C １－６ Alkilen-X ２ - [C １－６ Alkilen ｍ -POLY, -C ２－６ Alkenylene-X ２ - [C ２－６ Alkenirene] ｍ -POLY, or -C ２－６ Alkinylene-X ２ - [C ２－６ Alkinylene] ｍ - It is POLY, R ５０ Each alkylene, alkenylene, or alkynylene is a halogen, -CN, -NO ２ , -OH, -N(R １０ ) ２ , -C(O)N(R １０ ) ２ , -C(O)-, -C(S)-, -C(O)OCH ２ C ６ H ５ , -NHC(O)OCH ２ C ６ H ５ , C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ It may be substituted with one or more substituents selected from haloalkyl groups; R ５１ These are halogen, -CN, -NO ２ , -OH, -N(R １０ ) ２ , -C(O)N(R １０ ) ２ , -C(O)-, -C(S)-, -C(O)OCH ２ C ６ H ５ , -NHC(O)OCH ２ C ６ H ５ , C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ Selected independently from haloalkyl groups; X １ and X ２ is -N(R １０ )-, -C(O)-, and -N(R １０ ) Selected independently from C(O)-; R １０ In each appearance, hydrogen and C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ Selected independently from haloalkyl groups; POLY is a water-soluble polymer; n is an integer selected from 0, 1, 2, and 3; m is an integer selected from 0 and 1; p is an integer selected from 0 and 1; Su is a hexasaccharide monosaccharide; D is the drug part; RL is a reactive linker group residue.

2. L １ ga-C １－６ Alkylene- is; Y is -X １ -C １－６ alkylene-[X １ -C １－６ alkylene] ｎ -X １ -, -X １ -C ２－６ alkenylene-[X １ -C ２－６ alkenylene] ｎ -X １ -, or -X １ -C ２－６ alkynylene-[X １ -C ２－６ alkynylene] ｎ -X １ -, and at least one alkylene, alkenylene, or alkynylene of Y is substituted with one or more substituents selected from R ５０ ; R ５０ However, -C １－６ Alkilen-X ２ - [C １－６ Alkilen ｍ -POLY, -C ２－６ Alkenylene-X ２ - [C ２－６ Alkenirene] ｍ -POLY, or -C ２－６ Alkinylene-X ２ - [C ２－６ Alkinylene] ｍ - It is POLY, R ５０ Each alkylene, alkenylene, or alkynylene is a halogen, -CN, -NO ２ , -OH, -N(R １０ ) ２ , -C(O)N(R １０ ) ２ , -C(O)-, -C(S)-, -C(O)OCH ２ C ６ H ５ , -NHC(O)OCH ２ C ６ H ５ , C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ It may be substituted with one or more substituents selected from haloalkyl groups; X １ and X ２ However, -C(O)- and -N(R １０ ) Selected independently from C(O)-; R １０ However, in each appearance, hydrogen, C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ Selected independently from haloalkyl groups; POLY is a water-soluble polymer; n is an integer selected from 0, 1, 2, and 3; m is an integer selected from 0 and 1; Su is a monosaccharide in the hexasaccharide form; D is the drug part; The compound according to claim 1, wherein RL is a reactive linker group residue.

3. The compound of formula (I) is the compound of formula (IA) 【Chemistry 2】 The compound or salt according to claim 1, or a pharmaceutically acceptable salt thereof.

4. L １ ga-C １－３ The compound or salt according to claim 1, which is alkylene-.

5. The compound or salt according to claim 1, wherein p is 1.

6. Y is -X １ -C １－６ Alkylene-[X １ -C １－６ Alkilen ｎ -X １ - and at least one alkylene of Y is R ５０ The compound or salt according to claim 1, which is substituted with one or more substituents selected from the following.

7. Y is -X １ -C １－４ Alkylene-[X １ -C １－４ Alkilen ｎ -X １ - and at least one alkylene of Y is R ５０ The compound or salt according to claim 1, which is substituted with one or more substituents selected from the following.

8. Y is -X １ -C １－４ Alkilen-X １ -C １－４ Alkilen-X １ -C １－４ Alkilen-X １ - and at least one alkylene of Y is R ５０ The compound or salt according to claim 7, which is substituted with one or more substituents selected from the following.

9. Y is -X １ -C １－４ Alkilen-X １ -C １－４ Alkilen-X １ - and at least one alkylene of Y is R ５０ The compound or salt according to claim 7, which is substituted with one or more substituents selected from the following.

10. The compound or salt according to claim 1, wherein p is 0.

11. Y is -X １ -C １－６ Alkylene-[X １ -C １－６ Alkilen ｎ - and at least one alkylene of Y is R ５０ It is substituted with one or more substituents selected from, and the alkylene of Y is R ５１ The compound or salt according to claim 1, which may be substituted with one or more substituents selected from the following.

12. Y is -X １ -C １－４ Alkylene-[X １ -C １－４ Alkilen ｎ - and at least one alkylene of Y is R ５０ It is substituted with one or more substituents selected from, and the alkylene of Y is R ５１ The compound or salt according to claim 1, which may be substituted with one or more substituents selected from the following.

13. Y is -X １ -C １－４ Alkilen-X １ -C １－４ Alkilen-X １ -C １－４ It is an alkylene, and at least one alkylene of Y is R ５０ It is substituted with one or more substituents selected from, and the alkylene of Y is R ５１ The compound or salt according to claim 12, which may be substituted with one or more substituents selected from.

14. R ５１ However, halogen, -CN, -NO ２ -OH, -NH ２ , -C(O)NH ２ A compound or salt according to claim 1, independently selected from and -C(O)-.

15. R ５０ ga-C １－６ Alkilen-X ２ - [C １－６ Alkilen ｍ - It is POLY, R ５０ Each alkylene is a halogen, -CN, -NO ２ , -OH, -N(R １０ ) ２ , -C(O)N(R １０ ) ２ , -C(O)-, -C(S)-, -C(O)OCH ２ C ６ H ５ , -NHC(O)OCH ２ C ６ H ５ , C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ The compound or salt according to claim 1, which may be substituted with one or more substituents selected from haloalkyl groups.

16. The compound or salt according to claim 1, wherein m is 1.

17. The compound or salt according to claim 1, wherein POLY is polyethylene glycol (PEG), methoxypolyethylene glycol (mPEG), poly(propylene glycol) (PPG), copolymer of ethylene glycol and propylene glycol, poly(oxyethylated polyol), poly(olefin alcohol), poly(vinylpyrrolidone), 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.

18. The compound or salt according to claim 17, wherein POLY comprises polyethylene glycol (PEG) or methoxypolyethylene glycol (mPEG).

19. POLY 【Transformation 3】 And, 【Chemistry 4】 The compound or salt according to claim 1, wherein n1 represents a bond to the remainder of the compound, and n1 is an integer from 1 to 20.

20. The compound or salt according to claim 19, wherein n1 is an integer between 5 and 15.

21. The compound or salt according to claim 1, wherein RL comprises an alkyne, cyclooctin, strained alkene, tetrazine, methylcyclopropene, thiol, para-acetyl-phenylalanine residue, oxyamine, maleimide, or azide.

22. RL is 【Transformation 5】 , -N ３ , -NH ２ Selected from the group consisting of , and -SH; R Ｔ C １－６ It is alkyl; 【Transformation 6】 The compound or salt according to claim 1, wherein represents bonding to the remainder of the compound.

23. R Ｔ The compound or salt according to claim 22, wherein is methyl, ethyl, or propyl.

24. R Ｔ The compound or salt according to claim 23, wherein is methyl.

25. RL is 【Transformation 7】 A compound or salt according to claim 22, selected from the group consisting of the following.

26. RL 【Transformation 8】 The compound or salt according to claim 22.

27. Su 【Chemistry 9】 And, 【Chemistry 10】 The compound or salt according to claim 1, wherein represents bonding to the remainder of the compound.

28. Su 【Chemistry 11】 And, 【Chemistry 12】 The compound or salt according to claim 1, wherein represents bonding to the remainder of the compound.

29. The compound or salt according to claim 1, wherein D is an immunomodulatory payload.

30. The compound or salt according to claim 29, wherein the immunomodulatory payload is an agonist of interferon gene stimulating factor (STING), Toll-like receptor 7 (TLR7), Toll-like receptor 7 / 8 (TLR7 / 8), or Toll-like receptor 8 (TLR8).

31. The compound or salt according to claim 30, wherein the STING agonist is selected from the group consisting of small molecule agonists of the STING pathway, antibodies that activate STING activity, recombinant proteins that activate the STING pathway, TTI-10001, DMXAA (ASA404), CDN, c-Z-GMP, 2'3'-cGAMP, MK-1454, ADU-S100 (MIW815), SB11285, ADU-V19, IACS-8779, IACS-8803, IMSA101, non-CDN, E7766, MK-2118, diABZI, MSA-2, JNJ-'6196, bacterial vectors, SYNB1891, and STACT.

32. The compound or salt according to claim 30, wherein the agonist of Toll-like receptor 7 (TLR7) is selected from the group consisting of GS-986, PRTX-007, PRX-034, S-34240, MBS-8, and APR-002.

33. Toll-like receptor 7 / 8 (TLR7 / 8) agonists include imiquimod (R837), reximod (R848), 852-A (PF-4878691), besatrimod (GS-9620), AZD8848, motlimod (VTX-2337), sergantrimod (GS-9688), NKTR-262, RG-7854 (RO7020531), DSP-0509, BDB-001, and BDC-1001. A compound or salt according to claim 30, selected from the group consisting of LHC-165, SHR-2150, JNJ-4964 (TQ-73334), RO-7119929, DN-1508052, VTX-1463, BNT-411 (SC1), APR-003, ALT-702, TRANSCON, VX-001, SNAPvax, R848-HA, SM360320, and GSK2245035.

34. The compound or salt according to claim 30, wherein the Toll-like receptor 8 (TLR8) agonist is selected from the group consisting of SBT-6050, SBT-6290, and ZM-TLR8 agonists.

35. The compound or salt according to claim 1, wherein D is a cytotoxic payload.

36. The compound or salt according to claim 35, wherein the cytotoxic payload is a tubulin inhibitor, a DNA topoisomerase I inhibitor, or a DNA topoisomerase II inhibitor.

37. The compound or salt according to claim 1, wherein D is selected from the group consisting of hemiastalin, camptothecin, anthracycline, PNU-159682, and EDA PNU-159682 derivatives.

38. The compound or salt according to claim 1, wherein D is hemiastarin, exatecan, PNU-159682, or an EDA PNU-159682 derivative. 【Request Item 39】 【Chemistry 13-1】 【Chemistry 13-2】 【Chemistry 13-3】 The compound or salt according to claim 1, or selected from any one of the pharmaceutically acceptable salts thereof. 【Request Item 40】 【Chemistry 14】 The compound or salt according to claim 1, or selected from any one of the pharmaceutically acceptable salts thereof. 【Request Item 41】 【Chemistry 15-1】 【Chemistry 15-2】 【Chemistry 15-3】 The compound or salt according to claim 1, or selected from any one of the pharmaceutically acceptable salts thereof. 【Request Item 42】 【Chemistry 16-1】 【Chemistry 16-2】 The compound or salt according to claim 1, or selected from any one of the pharmaceutically acceptable salts thereof. 【Request Item 43】 【Chemistry 17-1】 【Chemistry 17-2】 The compound or salt according to claim 1, or selected from any one of the pharmaceutically acceptable salts thereof. 【Request Item 44】 【Chemistry 18】 The compound or salt according to claim 1, or selected from any one of the pharmaceutically acceptable salts thereof. 【Request Item 45】 【Chemistry 19】 The compound or salt according to claim 1, or selected from any one of the pharmaceutically acceptable salts thereof.

46. The following chemical structure 【Chemistry 20】 A compound having, or a pharmaceutically acceptable salt thereof.

47. The following chemical structure 【Chemistry 21】 A compound having, or a pharmaceutically acceptable salt thereof.

48. A conjugate comprising the compound according to claim 1, or a pharmaceutically acceptable salt thereof, linked to a second compound.

49. Structure of Equation II 【Chemistry 22】 (In the formula, COMP is a residue of the second compound; L １ Ha-C １－６ Alkylene- is; Y is -X １ -C １－６ Alkylene-[X １ -C １－６ Alkilen ｎ - [X １ ] ｐ -, -X １ -C ２－６ Alkenylene-[X １ -C ２－６ Alkenirene] ｎ - [X １ ] ｐ -, or -X １ -C ２－６ Alkinylene-[X １ -C ２－６ Alkinylene] ｎ - [X １ ] ｐ - and at least one alkylene, alkenylene, or alkynylene of Y is R ５０ It is substituted with one or more substituents selected from; Alkilen, alkenylene, or alkynylene of Y is R ５１ They may be substituted with one or more substituents selected from; R ５０ is, -C １－６ Alkilen-X ２ - [C １－６ Alkilen ｍ -POLY, -C ２－６ Alkenylene-X ２ - [C ２－６ Alkenirene] ｍ -POLY, or -C ２－６ Alkinylene-X ２ - [C ２－６ Alkinylene] ｍ - It is POLY, R ５０ Each alkylene, alkenylene, or alkynylene is a halogen, -CN, -NO ２ , -OH, -N(R １０ ) ２ , -C(O)N(R １０ ) ２ , -C(O)-, -C(S)-, -C(O)OCH ２ C ６ H ５ , -NHC(O)OCH ２ C ６ H ５ , C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ It may be substituted with one or more substituents selected from haloalkyl groups; R ５１ These are halogen, -CN, -NO ２ , -OH, -N(R １０ ) ２ , -C(O)N(R １０ ) ２ , -C(O)-, -C(S)-, -C(O)OCH ２ C ６ H ５ , -NHC(O)OCH ２ C ６ H ５ , C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ Selected independently from haloalkyl groups; X １ and X ２ is -N(R １０ )-, -C(O)-, and -N(R １０ ) Selected independently from C(O)-; R １０ In each appearance, hydrogen and C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ Selected independently from haloalkyl groups; POLY is a water-soluble polymer; n is an integer selected from 0, 1, 2, and 3; m is an integer selected from 0 and 1; p is an integer selected from 0 and 1; Su is a hexasaccharide monosaccharide; D is the drug part; RL is a reactive linker group residue. The conjugate according to claim 48.

50. COMP is the residue of the second compound; L １ ga-C １－６ Alkylene- is; Y is -X １ -C １－６ Alkylene-[X １ -C １－６ Alkilen ｎ -X １ -, -X １ -C ２－６ Alkenylene-[X １ -C ２－６ Alkenirene] ｎ -X １ -, or -X １ -C ２－６ Alkinylene-[X １ -C ２－６ Alkinylene] ｎ -X １ - and at least one alkylene, alkenylene, or alkynylene of Y is R ５０ It is substituted with one or more substituents selected from; R ５０ However, -C １－６ Alkilen-X ２ - [C １－６ Alkilen ｍ -POLY, -C ２－６ Alkenylene-X ２ - [C ２－６ Alkenirene] ｍ -POLY, or -C ２－６ Alkinylene-X ２ - [C ２－６ Alkinylene] ｍ - It is POLY, R ５０ Each alkylene, alkenylene, or alkynylene is a halogen, -CN, -NO ２ , -OH, -N(R １０ ) ２ , -C(O)N(R １０ ) ２ , -C(O)-, -C(S)-, -C(O)OCH ２ C ６ H ５ , -NHC(O)OCH ２ C ６ H ５ , C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ It may be substituted with one or more substituents selected from haloalkyl groups; X １ and X ２ However, -C(O)- and -N(R １０ ) Selected independently from C(O)-; R １０ However, in each appearance, hydrogen, C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ Selected independently from haloalkyl groups; POLY is a water-soluble polymer; n is an integer selected from 0, 1, 2, and 3; m is an integer selected from 0 and 1; Su is a monosaccharide in the hexasaccharide form; D is the drug part; The conjugate according to claim 49, wherein RL is a reactive linker group residue.

51. The conjugate according to claim 49, wherein COMP is a polypeptide residue.

52. The conjugate according to claim 49, wherein COMP is a residue of an antibody.

53. The conjugate according to claim 49, wherein COMP is a residue of the antibody chain.

54. The conjugate of equation (II) is equation (IIA). 【Chemistry 23】 The conjugate according to claim 49, which is made by

55. L １ ga-C １－３ The conjugate according to claim 49, wherein the conjugate is alkylene.

56. The conjugate according to claim 49, wherein p is 1.

57. Y is -X １ -C １－６ Alkylene-[X １ -C １－６ Alkilen ｎ -X １ - and at least one alkylene of Y is R ５０ The conjugate according to claim 50, which is substituted with one or more substituents selected from the following.

58. Y is -X １ -C １－４ Alkylene-[X １ -C １－４ Alkilen ｎ -X １ - and at least one alkylene of Y is R ５０ The conjugate according to claim 50, which is substituted with one or more substituents selected from the following.

59. Y is -X １ -C １－４ Alkilen-X １ -C １－４ Alkilen-X １ -C １－４ Alkilen-X １ - and at least one alkylene of Y is R ５０ The conjugate according to claim 58, which is substituted with one or more substituents selected from the following.

60. Y is -X １ -C １－４ Alkilen-X １ -C １－４ Alkilen-X １ - and at least one alkylene of Y is R ５０ The conjugate according to claim 58, which is substituted with one or more substituents selected from the following.

61. The conjugate according to claim 49, wherein p is 0.

62. Y is -X １ -C １－６ Alkylene-[X １ -C １－６ Alkilen ｎ - and at least one alkylene of Y is R ５０ It is substituted with one or more substituents selected from, and the alkylene of Y is R ５１ The conjugate according to claim 49, which may be substituted with one or more substituents selected from the following.

63. Y is -X １ -C １－４ Alkylene-[X １ -C １－４ Alkilen ｎ - and at least one alkylene of Y is R ５０ It is substituted with one or more substituents selected from, and the alkylene of Y is R ５１ The conjugate according to claim 49, which may be substituted with one or more substituents selected from the following.

64. Y is -X １ -C １－４ Alkilen-X １ -C １－４ Alkilen-X １ -C １－４ It is an alkylene, and at least one alkylene of Y is R ５０ It is substituted with one or more substituents selected from, and the alkylene of Y is R ５１ The conjugate according to claim 63, which may be substituted with one or more substituents selected from the following.

65. R ５１ However, halogen, -CN, -NO ２ -OH, -NH ２ , -C(O)NH ２ The conjugate according to claim 49, which is independently selected from and -C(O)-.

66. R ５０ ga-C １－６ Alkilen-X ２ - [C １－６ Alkilen ｍ - It is POLY, R ５０ Each alkylene is a halogen, -CN, -NO ２ , -OH, -N(R １０ ) ２ , -C(O)N(R １０ ) ２ , -C(O)-, -C(S)-, -C(O)OCH ２ C ６ H ５ , -NHC(O)OCH ２ C ６ H ５ , C １－１０ Alkyl, C ２－１０ Alkenil, C ２－１０ Alkinyl, C ３－１２ Carbon rings, 3- to 12-membered heterocycles, and C １－１０ The conjugate according to claim 49, which may be substituted with one or more substituents selected from haloalkyl groups.

67. The conjugate according to claim 49, wherein m is 1.

68. The conjugate according to claim 49, wherein POLY is polyethylene glycol (PEG), methoxypolyethylene glycol (mPEG), poly(propylene glycol) (PPG), copolymer of ethylene glycol and propylene glycol, poly(oxyethylated polyol), poly(olefin alcohol), poly(vinylpyrrolidone), 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.

69. The conjugate according to claim 66, wherein POLY comprises polyethylene glycol (PEG) or methoxypolyethylene glycol (mPEG).

70. POLY 【Chemistry 24】 And, 【Chemistry 25】 The conjugate according to claim 49, wherein n represents bonding to the remainder of the compound, and n1 is an integer from 1 to 20.

71. The conjugate according to claim 70, wherein n1 is an integer between 5 and 15.

72. The conjugate according to claim 49, wherein RL comprises an alkyne, cyclooctin, strained alkene, tetrazine, thiol, para-acetyl-phenylalanine residue, oxyamine, amine, maleimide, or azide.

73. RL is 【Chemistry 26】 Selected from the group consisting of; R Ｔ C １－６ Alkyl 【Chemistry 27】 This represents bonding to the remainder of the compound. The conjugate according to claim 49.

74. R Ｔ The conjugate according to claim 49, wherein is methyl, ethyl, or propyl.

75. R Ｔ The conjugate according to claim 49, wherein is methyl.

76. RL is 【Chemistry 28】 A conjugate according to claim 73, selected from the group consisting of the following.

77. RL is 【Chemistry 29】 A conjugate according to claim 76, selected from the group consisting of the following.

78. Su 【Transformation 30】 And, 【Chemistry 31】 The conjugate according to claim 49, wherein represents bonding to the remainder of the compound.

79. Su 【Chemistry 32】 And, 【Transformation 33】 The conjugate according to claim 49, wherein represents bonding to the remainder of the compound.

80. The conjugate according to claim 49, wherein D is an immunomodulatory payload.

81. The conjugate according to claim 80, wherein the immunomodulatory payload is an agonist of interferon gene stimulating factor (STING), Toll-like receptor 7 (TLR7), Toll-like receptor 7 / 8 (TLR7 / 8), or Toll-like receptor 8 (TLR8).

82. The conjugate according to claim 81, wherein the STING agonist is selected from the group consisting of a small molecule agonist of the STING pathway, an antibody that activates STING activity, a recombinant protein that activates the STING pathway, TTI-10001, DMXAA (ASA404), CDN, c-Z-GMP, 2'3'-cGAMP, MK-1454, ADU-S100 (MIW815), SB11285, ADU-V19, IACS-8779, IACS-8803, IMSA101, non-CDN, E7766, MK-2118, diABZI, MSA-2, JNJ-'6196, a bacterial vector, SYNB1891, and STACT.

83. The conjugate according to claim 81, wherein the agonist of Toll-like receptor 7 (TLR7) is selected from the group consisting of GS-986, PRTX-007, PRX-034, S-34240, MBS-8, and APR-002.

84. Toll-like receptor 7 / 8 (TLR7 / 8) agonists include imiquimod (R837), reximod (R848), 852-A (PF-4878691), besatrimod (GS-9620), AZD8848, motlimod (VTX-2337), sergantrimod (GS-9688), NKTR-262, RG-7854 (RO7020531), DSP-0509, BDB-001, and BDC-1001. The conjugate according to claim 81, selected from the group consisting of LHC-165, SHR-2150, JNJ-4964 (TQ-73334), RO-7119929, DN-1508052, VTX-1463, BNT-411 (SC1), APR-003, ALT-702, TRANSCON, VX-001, SNAPvax, R848-HA, SM360320, and GSK2245035.

85. The conjugate according to claim 81, wherein the Toll-like receptor 8 (TLR8) agonist is selected from the group consisting of SBT-6050, SBT-6290, and ZM-TLR8 agonists.

86. The conjugate according to claim 49, wherein D is a cytotoxic payload.

87. The conjugate according to claim 86, wherein the cytotoxic payload is a tubulin inhibitor, a DNA topoisomerase I inhibitor, or a DNA topoisomerase II inhibitor.

88. The conjugate according to claim 49, wherein D is selected from the group consisting of hemiastalin, camptothecin, anthracycline, PNU-159682, and EDA PNU-159682 derivatives.

89. The conjugate according to claim 49, wherein D is hemiasterin, exatecan, PNU-159682, or an EDA PNU-159682 derivative.

90. The compound, 【Chemistry 34-1】 【Chemistry 34-2】 【Chemistry 34-3】 【Chemistry 34-4】 【Chemistry 34-5】 【Chemistry 34-6】 ; or selected from any one of the pharmaceutically acceptable salts thereof, 【Chemistry 35】 The conjugate according to claim 49, wherein is the second compound residue.

91. The compound, 【Chemistry 36-1】 【Chemistry 36-2】 ; or selected from any one of the pharmaceutically acceptable salts thereof, 【Chemistry 37】 The conjugate according to claim 50, wherein is the second compound residue.

92. The compound, 【Chemistry 38-1】 【Chemistry 38-2】 【Chemistry 38-3】 【Chemistry 38-4】 【Chemistry 38-5】 【Chemistry 38-6】 ; or selected from any one of the pharmaceutically acceptable salts thereof, 【Chemistry 39】 The conjugate according to claim 50, wherein is the second compound residue. 【Request Item 93】 【Chemistry 40-1】 【Chemistry 40-2】 , or selected from any one of their pharmaceutically acceptable salts, The conjugate according to claim 49, wherein COMP is a residue of the second compound. 【Request Item 94】 【Chemistry 41-1】 【Chemistry 41-2】 , or selected from any one of their pharmaceutically acceptable salts, The conjugate according to claim 49, wherein COMP is a residue of the second compound. 【Request Item 95】 【Chemistry 42】 , or selected from any one of their pharmaceutically acceptable salts, The conjugate according to claim 49, wherein COMP is a residue of the second compound. 【Request Item 96】 【Chemistry 43】 , or selected from any one of their pharmaceutically acceptable salts, The conjugate according to claim 49, wherein COMP is a residue of the second compound.

97. The following chemical structure 【Chemistry 44】 ; and any one of their pharmaceutically acceptable salts (In the formula, 【Chemistry 45】 (This is a residue of the second compound.) A conjugate that possesses this property.

98. The following chemical structure 【Chemistry 46】 ; and any one of their pharmaceutically acceptable salts (In the formula, 【Chemistry 47】 (This is a residue of the second compound.) A conjugate that possesses this property.

99. A pharmaceutical composition comprising a compound according to any one of claims 1 to 47 or a conjugate according to any one of claims 48 to 98, and a pharmaceutically acceptable excipient, carrier, or diluent.

100. A pharmaceutical composition for use in treatment, comprising the pharmaceutical composition according to claim 99, or the compound according to any one of claims 1 to 47, or the conjugate according to any one of claims 48 to 98.

101. A pharmaceutical composition for use in the treatment of cancer, comprising the pharmaceutical composition according to claim 99, or the compound according to any one of claims 1 to 47, or the conjugate according to any one of claims 48 to 98.

102. A pharmaceutical composition for use as a pharmaceutical, comprising the pharmaceutical composition according to claim 99, or the compound according to any one of claims 1 to 47, or the conjugate according to any one of claims 48 to 98.

103. Use of a compound according to any one of claims 1 to 47, a conjugate according to any one of claims 48 to 98, or a pharmaceutical composition according to claim 99 in the production of a pharmaceutical that inhibits tubulin polymerization.

104. Use of a compound according to any one of claims 1 to 47, a conjugate according to any one of claims 48 to 98, or a pharmaceutical composition according to claim 99 in the manufacture of a pharmaceutical that reduces cell proliferation.

105. Use of a compound according to any one of claims 1 to 47, a conjugate according to any one of claims 48 to 98, or a pharmaceutical composition according to claim 99 in the manufacture of a pharmaceutical for treating cancer.

106. The use according to claim 105, 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, Her2-overexpressing breast cancer, triple-negative breast cancer, lymphoma, large cell lymphoma, diffuse mixed histiocytic and lymphocytic lymphoma, follicular B-cell lymphoma, colon cancer, colon adenocarcinoma, colorectal adenocarcinoma, melanoma, prostate cancer, or multiple myeloma.

107. A method for producing a conjugate, comprising the step of contacting a compound according to any one of claims 1 to 47 with a second compound under conditions suitable for conjugating the compound according to any one of claims 1 to 47 with a second compound, wherein the second compound comprises an alkyne, cyclooctyne, strained alkene, tetrazine, methylcyclopropene, thiol, maleimide, carbonyl, amine, oxyamine, or azide.

108. The method according to claim 107, wherein the second compound comprises tetrazine; and RL comprises a strained alkene.

109. RL 【Chemistry 48】 The method according to claim 107.

110. The method according to claim 107, wherein the second compound comprises an azide; and RL comprises an alkyne.

111. RL 【Chemistry 49】 The method according to claim 107.

112. The method according to claim 107, wherein the second compound comprises an alkyne; and RL comprises an azide.

113. The method according to claim 107, wherein the second compound comprises a strained alkene; and RL comprises tetrazine.

114. The method according to claim 107, wherein the second compound comprises a thiol; and RL comprises maleimide.

115. RL [Transformation 50] The method according to claim 107.

116. The method according to claim 107, wherein the second compound comprises maleimide; and RL comprises a thiol.

117. The second compound is 【Chemistry 51】 The method according to claim 107, including the method described in claim 107.

118. The method according to claim 107, wherein the second compound comprises a carbonyl group; and RL comprises an oxyamine group.

119. RL 【Chemistry 52】 The method according to claim 107.

120. The second compound is 【Chemistry 53】 The method according to claim 107, including the method described in claim 107.

121. The method according to claim 107, wherein the second compound comprises an oxyamine; and RL comprises a carbonyl.

122. RL 【Chemistry 54】 The method according to claim 107.

123. The second compound is 【Transformation 55】 The method according to claim 107, including the method described in claim 107.

124. The method according to claim 107, wherein the second compound is a polypeptide.

125. The method according to claim 107, wherein the second compound is an antibody.

126. The method according to claim 107, wherein the second compound is an antibody chain.

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