Compositions and uses thereof

Abstract

Provided herein are compounds of Formula (I *): # imgabs0 # or a pharmaceutically acceptable salt thereof, wherein the variables of Formula (I *) are as defined herein. Such compounds may be useful as anticancer agents.

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 377,185, filed on September 26, 2022, and U.S. Provisional Patent Application No. 63 / 384,530, filed on November 21, 2022, each of which is hereby incorporated by reference in its entirety. Background Art

[0003] Currently, small cytotoxic molecules for antibody-drug conjugates can include camptothecin derivatives, which have antitumor effects by inhibiting topoisomerase I. Camptothecin derivatives can be used in antibody-drug conjugates (ADCs). However, there is still a need to further develop camptothecin derivatives and ADC drugs with better efficacy and / or higher safety. Summary of the Invention

[0004] In one aspect, the present disclosure provides a compound represented by formula (I*),

[0005]

[0006] or a pharmaceutically acceptable salt thereof, wherein;

[0007] L and M are linkers;

[0008] n and u are selected from 0 and 1;

[0009] R 1A is selected from hydrogen and C 1-6 alkyl;

[0010] R 1B is optionally substituted C 1-6 alkyl, wherein C 1-6 alkyl is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NH2, and oxo;

[0011] wherein when u is 1, R 1B is absent or is -C 1-6 alkylene-O-; and

[0012] wherein either u or n is 1.

[0013] In some embodiments, formula (I*) is represented as

[0014]

[0015] or a pharmaceutically acceptable salt thereof;

[0016] R 1A is selected from C 1-6 alkyl;

[0017] R 1B selected from C 1-6 alkyl; and

[0018] L is a linker.

[0019] In some embodiments, formula (I*) or formula (I) is represented as

[0020]

[0021] or a pharmaceutically acceptable salt thereof.

[0022] In some embodiments, the linker (e.g., L) is L 1 -L 2 -L 3 -L 4 -(L 5 ) m -R 2 ; and

[0023] L 1 selected from C 1-6 alkylene;

[0024] L 2 selected from optionally substituted C 5-6 carbocycle, wherein C 5-6 carbocycle is optionally substituted with one or more substituents independently selected from R 3 ;

[0025] L 3 selected from residues comprising 1 to 7 amino acids;

[0026] L 4 selected from optionally substituted C 1-6 alkylene, wherein C 1-6 alkylene is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0027] L 5 selected from (-O-CH2-CH2-) q ;

[0028] R 2 selected from optionally substituted 5- to 6-membered heterocycles, wherein the 5- to 6-membered heterocycle is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C1-10 alkyl, C 2-10 alkenyl, C 2-10 substituted by a substituent of alkynyl;

[0029] Each R 3 is independently selected from sugars;

[0030] m is selected from 0 and 1; and

[0031] q is selected from 1 to 10.

[0032] In some embodiments, formula (I*) is represented as

[0033]

[0034] or a pharmaceutically acceptable salt thereof, wherein;

[0035] R 1A is selected from hydrogen and C 1-6 alkyl;

[0036] R 1B is absent or is -C 1-6 alkylene-O-;

[0037] M is (L 2 ) x -(L 2B ) z -(L 2C ) y -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 ;

[0038] L 2 is selected from C 1-6 alkylene;

[0039] L 2B is selected from (NR 4 ) t C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted by one or more R 5 substituents;

[0040] L 2C is selected from C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted by one or more R 6 substituents;

[0041] L 3 is selected from residues comprising 1 to 7 amino acids;

[0042] L 4 selected from optionally substituted C 1-6 alkylene, wherein C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0043] L 5 selected from (O-CH2-CH2) q -(NR 3 ) s ;

[0044] L 6 selected from optionally substituted C 1-6 alkylene, wherein C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0045] L 7 selected from C 5-6 carbocycle;

[0046] R 2 selected from optionally substituted 5- to 6-membered heterocycles, wherein the 5- to 6-membered heterocycles are optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0047] R 3 selected from hydrogen and C 1-6 alkyl;

[0048] R 4 selected from hydrogen and C 1-6 alkyl optionally substituted by one or more SO2C 1-6 alkyl;

[0049] Each R 5 is independently selected from sugars;

[0050] Each R 6 is independently selected from halogen, -OH, -CN, -NO2, -NH2, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0051] m is selected from 0 and 1;

[0052] p is selected from 0 and 1;

[0053] q is selected from 0 to 8;

[0054] s is selected from 0 and 1;

[0055] t is selected from 0 and 1;

[0056] x is selected from 0 and 1;

[0057] y is selected from 0 and 1; and

[0058] z is selected from 0 and 1.

[0059] In another aspect, the present disclosure provides a compound of formula (III):

[0060]

[0061] or a pharmaceutically acceptable salt thereof, wherein;

[0062] R 1A is selected from C 1-6 alkyl;

[0063] R 1B is optionally substituted C 1-6 alkyl, wherein the C 1-6 alkyl is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NH2, and oxo;

[0064] L is a linker; and

[0065] Lg is a ligand.

[0066] In some embodiments, the ligand is selected from an antibody or an antigen-binding fragment thereof. In some cases, the ligand is selected from a chimeric antibody, a humanized antibody, and a fully human antibody.

[0067] In some embodiments, the compound is represented by formula (VII)

[0068]

[0069] or a pharmaceutically acceptable salt thereof.

[0070] In some embodiments, the compound is represented as

[0071]

[0072] or a pharmaceutically acceptable salt thereof, wherein the DAR is selected from about 1 - 8. In some cases, the DAR is 4.

[0073] In some embodiments, the compound is represented by formula (VIII)

[0074]

[0075] or a pharmaceutically acceptable salt thereof, wherein the DAR is selected from about 1 - 8, and Lg is a ligand. In some cases, the DAR is about 4.

[0076] In some embodiments, the pharmaceutical composition comprises a compound or salt of formula (I*), formula (I), formula (I - A), formula (I - B), formula (II), formula (II - A), formula (III), formula (III - A), formula (IV), formula (IV - A), formula (IV - B), formula (IV - C), formula (IV - D), formula (V), formula (VI), formula (VII), formula (VIII), and a pharmaceutically acceptable excipient.

[0077] In some embodiments, the present disclosure provides a method of treating a subject having a tumor, comprising administering to a subject in need thereof a compound or salt of formula (I*), formula (I), formula (I - A), formula (I - B), formula (II), formula (II - A), formula (III), formula (III - A), formula (IV), formula (IV - A), formula (IV - B), formula (IV - C), formula (IV - D), formula (V), formula (VI), formula (VII), formula (VIII), or a pharmaceutical composition thereof.

[0078] In some embodiments, the present disclosure provides a method of treating a subject having cancer, comprising administering to a subject in need thereof a compound or salt of formula (I*), formula (I), formula (I - A), formula (I - B), formula (II), formula (II - A), formula (III), formula (III - A), formula (IV), formula (IV - A), formula (IV - B), formula (IV - C), formula (IV - D), formula (V), formula (VI), formula (VII), formula (VIII), or a pharmaceutical composition thereof.

[0079] Incorporation by reference

[0080] All publications, patents, and patent applications mentioned in this specification are hereby incorporated by reference in their entirety as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent that any incorporated publication, patent, or patent application conflicts with the disclosure contained herein, this specification is intended to supersede and / or take precedence over any such conflicting material. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the invention will be obtained by reference to the following detailed description that sets forth exemplary embodiments in which the principles of the invention are utilized, as well as the accompanying drawings (also referred to herein as “figures” and “drawings”). In the drawings:

[0082] Figure 1 illustrates the stability of ADC-1 in mouse, cynomolgus monkey, and human plasma;

[0083] Figure 2 illustrates the cell binding of ADC-1 in Jeko-1 cells; and

[0084] Figure 3 illustrates the internalization of ADC-1 in Jeko-1 cells. DETAILED DESCRIPTION

[0085] The following description sets forth numerous exemplary configurations, methods, parameters, etc. However, it should be understood that such description is not intended to limit the scope of the disclosure, but is provided merely as a description of exemplary embodiments.

[0086] In the following description, certain specific details are set forth to provide a thorough understanding of various embodiments of the disclosure. However, one skilled in the art will understand that the disclosure may be practiced without these details.

[0087] DEFINITIONS

[0088] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications cited herein are incorporated by reference.

[0089] “Alkyl” refers to a straight-chain or branched-chain hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing no unsaturation, and preferably having from one to fifteen carbon atoms (i.e., C1-C 15 alkyl). In certain embodiments, alkyl contains from one to thirteen carbon atoms (i.e., C1-C 13(alkyl). In certain embodiments, the alkyl contains one to eight carbon atoms (i.e., C1-C8 alkyl). In other embodiments, the alkyl contains one to five carbon atoms (i.e., C1-C5 alkyl). In other embodiments, the alkyl contains one to four carbon atoms (i.e., C1-C4 alkyl). In other embodiments, the alkyl contains one to three carbon atoms (i.e., C1-C3 alkyl). In other embodiments, the alkyl contains one to two carbon atoms (i.e., C1-C2 alkyl). In other embodiments, the alkyl contains one carbon atom (i.e., C1 alkyl). In other embodiments, the alkyl contains five to fifteen carbon atoms (i.e., C5-C 15 (alkyl). In other embodiments, the alkyl contains five to eight carbon atoms (i.e., C5-C8 alkyl). In other embodiments, the alkyl contains two to five carbon atoms (i.e., C2-C5 alkyl). In other embodiments, the alkyl contains three to five carbon atoms (i.e., C3-C5 alkyl). In certain embodiments, the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (isopropyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (isobutyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl is attached to the rest of the molecule by a single bond.

[0090] The term "C x-y " when used in combination with a chemical moiety (such as alkyl, alkenyl or alkynyl) means a group containing x to y carbons in the chain. For example, "C 1-6 alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group, including straight-chain and branched-chain alkyl groups containing 1 to 6 carbons. The term -C x-y alkylene- refers to a substituted or unsubstituted alkylene chain, where the alkylene chain has x to y carbon atoms. For example, -C 1-6 alkylene- can be selected from methylene, ethylene, propylene, butylene, pentylene and hexylene, any of which is optionally substituted.

[0091] "Alkoxy" refers to a group of the formula -O-alkyl bonded through an oxygen atom, where the alkyl is an alkyl chain as defined above.

[0092] "Alkenyl" refers to a straight-chain or branched-chain hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and preferably having two to twelve carbon atoms (i.e., C2-C 12(alkenyl). In certain embodiments, the alkenyl contains two to eight carbon atoms (i.e., C2-C8 alkenyl). In certain embodiments, the alkenyl contains two to six carbon atoms (i.e., C2-C6 alkenyl). In other embodiments, the alkenyl contains two to four carbon atoms (i.e., C2-C4 alkenyl). The alkenyl is attached to the remainder of the molecule by a single bond, such as ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, pent-1,4-dienyl, etc.

[0093] "Alkynyl" refers to a straight-chain or branched-chain hydrocarbon chain group consisting of only carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, and preferably having two to twelve carbon atoms (i.e., C2-C 12 alkynyl). In certain embodiments, the alkynyl contains two to eight carbon atoms (i.e., C2-C8 alkynyl). In other embodiments, the alkynyl contains two to six carbon atoms (i.e., C2-C6 alkynyl). In other embodiments, the alkynyl contains two to four carbon atoms (i.e., C2-C4 alkynyl). The alkynyl is attached to the remainder of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc.

[0094] The term "C x-y alkenyl" and "C x-y alkynyl" refer to substituted or unsubstituted unsaturated aliphatic groups, which are similar in length and possible substitution to the above-mentioned alkyl groups, but contain at least one double bond or triple bond respectively. The term -C x-y alkenylene- refers to a substituted or unsubstituted alkenylene chain, where there are x to y carbons in the alkenylene chain. For example, -C 2-6 alkenylene- can be selected from vinylene, propenylene, butenylene, pentenylene, and hexenylene, any of which is optionally substituted. The alkenylene chain can have one double bond or more than one double bond in the alkenylene chain. The term -C x-y alkynylene- refers to a substituted or unsubstituted alkynylene chain with x to y carbons in the alkynylene chain. For example, -C 2-6 alkynylene- can be selected from ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any of which is optionally substituted. The alkynylene chain can have one triple bond or more than one triple bond in the alkynylene chain.

[0095] "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain that connects the remainder of the molecule to a radical group consisting only of carbon and hydrogen, contains no unsaturation, and preferably has one to twelve carbon atoms, such as methylene, ethylene, propylene, n-butylene, etc. The alkylene chain is attached to the remainder of the molecule by a single bond and to the radical group by a single bond. The attachment points of the alkylene chain to the remainder of the molecule and to the radical group can be through any two carbons in the chain. In certain embodiments, the alkylene contains one to ten carbon atoms (i.e., C1-C 10 alkylene). In certain embodiments, the alkylene contains one to eight carbon atoms (i.e., C1-C8 alkylene). In other embodiments, the alkylene contains one to five carbon atoms (i.e., C 1- C5 alkylene). In other embodiments, the alkylene contains one to four carbon atoms (i.e., C1-C4 alkylene). In other embodiments, the alkylene contains one to three carbon atoms (i.e., C1-C3 alkylene). In other embodiments, the alkylene contains one to two carbon atoms (i.e., C1-C2 alkylene). In other embodiments, the alkylene contains one carbon atom (i.e., C1 alkylene). In other embodiments, the alkylene contains five to eight carbon atoms (i.e., C5-C8 alkylene). In other embodiments, the alkylene contains two to five carbon atoms (i.e., C2-C5 alkylene). In other embodiments, the alkylene contains three to five carbon atoms (i.e., C3-C5 alkylene).

[0096] "Alkenylene" or "alkenylene chain" refers to a straight or branched divalent hydrocarbon chain that connects the remainder of the molecule to a radical group consisting only of carbon and hydrogen, contains at least one carbon-carbon double bond, and preferably has two to twelve carbon atoms. The alkenylene chain is attached to the remainder of the molecule by a single bond and to the radical group by a single bond. The attachment points of the alkenylene chain to the remainder of the molecule and to the radical group can be through any two carbon atoms in the chain. In certain embodiments, the alkenylene contains two to ten carbon atoms (i.e., C2-C 10 alkenylene). In certain embodiments, the alkenylene contains two to eight carbon atoms (i.e., C2-C8 alkenylene). In other embodiments, the alkenylene contains two to five carbon atoms (i.e., C2-C5 alkenylene). In other embodiments, the alkenylene contains two to four carbon atoms (i.e., C2-C4 alkenylene). In other embodiments, the alkenylene contains two to three carbon atoms (i.e., C2-C3 alkenylene). In other embodiments, the alkenylene contains two carbon atoms (i.e., C2 alkenylene). In other embodiments, the alkenylene contains five to eight carbon atoms (i.e., C5-C8 alkenylene). In other embodiments, the alkenylene contains three to five carbon atoms (i.e., C3-C5 alkenylene).

[0097] "Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain that attaches the remainder of the molecule to a radical group consisting only of carbon and hydrogen, contains at least one carbon-carbon triple bond, and preferably has two to twelve carbon atoms. The alkynylene chain is attached to the remainder of the molecule by a single bond and to the radical group by a single bond. The points of attachment of the alkynylene chain to the remainder of the molecule and to the radical group can be through any two carbon atoms in the chain. In certain embodiments, the alkynylene contains two to ten carbon atoms (i.e., C2-C 10 alkynylene). In certain embodiments, the alkynylene contains two to eight carbon atoms (i.e., C2-C8 alkynylene). In other embodiments, the alkynylene contains two to five carbon atoms (i.e., C2-C5 alkynylene). In other embodiments, the alkynylene contains two to four carbon atoms (i.e., C2-C4 alkynylene). In other embodiments, the alkynylene contains two to three carbon atoms (i.e., C2-C3 alkynylene). In other embodiments, the alkynylene contains two carbon atoms (i.e., C2 alkynylene). In other embodiments, the alkynylene contains five to eight carbon atoms (i.e., C5-C8 alkynylene). In other embodiments, the alkynylene contains three to five carbon atoms (i.e., C3-C5 alkynylene).

[0098] "Aryl" refers to a radical obtained by removing a hydrogen atom from a ring carbon atom of an aromatic monocyclic or aromatic polycyclic hydrocarbon ring system. The aromatic monocyclic or aromatic polycyclic hydrocarbon ring system contains only hydrogen and carbon, and has 5 to 18 carbon atoms, wherein at least one ring in the ring system is aromatic, i.e., according to Hückel's theory, it contains a cyclic, delocalized (4n + 2)π - electron system. Ring systems from which aryl is derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetrahydronaphthalene, and naphthalene.

[0099] "Aralkyl" refers to a radical of the formula -R c -aryl, where R c is an alkylene chain as defined above, such as methylene, ethylene, etc.

[0100] "Arenyl" refers to a radical of the formula -R d -aryl, where R d is an alkenylene chain as defined above. "Arylalkynyl" refers to a radical of the formula -R e -aryl, where R e is an alkynylene chain as defined above.

[0101] "Carbocyclic ring" means a saturated ring, an unsaturated ring or an aromatic ring, wherein each atom of the ring is carbon. The carbocyclic ring can include a 3- to 10-membered monocyclic ring, a 6- to 12-membered bicyclic ring and a 6- to 12-membered bridged ring. Each ring of the bicyclic carbocyclic ring can be selected from a saturated ring, an unsaturated ring and an aromatic ring. An aromatic ring (such as phenyl) can be fused with a saturated ring or an unsaturated ring, such as cyclohexane, cyclopentane or cyclohexene. The definition of the carbocyclic ring includes any combination of saturated rings, unsaturated rings and aromatic rings (as long as the valence allows). Exemplary carbocyclic rings include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl and naphthyl. The bicyclic carbocyclic ring can be a fused, bridged or spiro ring system. In some cases, the spiro carbocyclic ring has at least two molecular rings, wherein there is only one common atom.

[0102] "Carbocyclene" means a divalent carbocyclic ring that attaches the remainder of the molecule to a radical group.

[0103] The term "unsaturated carbocyclic ring" means a carbocyclic ring having at least one degree of unsaturation and not containing an aromatic carbocyclic ring. Examples of unsaturated carbocyclic rings include cyclohexadiene, cyclohexene and cyclopentene.

[0104] "Cycloalkyl" means a fully saturated monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms, which includes a fused ring or a bridged ring system and preferably has three to twelve carbon atoms. In certain embodiments, the cycloalkyl contains three to ten carbon atoms. In other embodiments, the cycloalkyl contains five to seven carbon atoms. The cycloalkyl can be attached to the remainder of the molecule by a single bond. Examples of monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Examples of polycyclic cycloalkyls include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptyl), norbornenyl, decahydronaphthyl, 7,7-dimethylbicyclo[2.2.1]heptyl, etc.

[0105] "Cycloenyl" means an unsaturated non-aromatic monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms, which includes a fused ring or a bridged ring system, preferably has three to twelve carbon atoms, and contains at least one double bond. In certain embodiments, the cycloenyl contains three to ten carbon atoms. In other embodiments, the cycloenyl contains five to seven carbon atoms. The cycloenyl can be attached to the remainder of the molecule by a single bond. Examples of monocyclic cycloenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl.

[0106] "Cycloalkylalkyl" means a radical of the formula -R c -cycloalkyl, wherein R c is an alkylene chain as described above.

[0107] "Cycloalkylalkoxy" means a radical bonded through an oxygen atom of the formula -O-R c -cycloalkyl, wherein Rc is an alkylene chain as described above.

[0108] "Halogenated" or "halogen" refers to halogen substituents such as bromine, chlorine, fluorine, and iodine substituents.

[0109] As used herein, the term "haloalkyl" or "haloalkane" refers to an alkyl radical as defined above that is substituted with one or more halogen groups, such as trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. In some embodiments, the alkyl portion of the fluoroalkyl radical is optionally further substituted. Examples of halogen-substituted alkanes ("haloalkanes") include halomethanes (e.g., chloromethane, bromomethane, fluoromethane, iodomethane), dihalomethanes, and trihalomethanes (e.g., trichloromethane, tribromomethane, trifluoromethane, triiodomethane), 1-haloethanes, 2-haloethanes, 1,2-dihaloethanes, 1-halopropanes, 2-halopropanes, 3-halopropanes, 1,2-dihalopropanes, 1,3-dihalopropanes, 2,3-dihalopropanes, 1,2,3-trihalopropanes, and any other suitable combination of an alkane (or substituted alkane) and a halogen (e.g., chlorine, bromine, fluorine, iodine, etc.). When an alkyl group is substituted with more than one halogen group, each halogen can be independently selected, e.g., 1-chloro,2-fluoroethane.

[0110] "Fluoroalkyl" refers to an alkyl radical as defined above that is substituted with one or more fluorine groups. For example, trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc.

[0111] "Aminoalkyl" refers to an alkyl radical as defined above that is substituted with one or more amine radicals, such as propan-2-amine, butane-1,2-diamine, pentane-1,2,4-triamine, etc.

[0112] "Hydroxyalkyl" refers to an alkyl radical as defined above that is substituted with one or more hydroxy radicals, such as propan-1-ol, butane-1,4-diol, pentane-1,2,4-triol, etc.

[0113] "Alkoxyalkyl" refers to an alkyl radical as defined above that is substituted with one or more alkoxy radicals, such as methoxymethane, 1,3-dimethoxybutane, 1-methoxypropane, 2-ethoxypentane, etc.

[0114] "Cyanoalkyl" as used herein refers to an alkyl radical as defined above that is substituted with one or more cyano radicals, such as acetonitrile, 2-ethyl-3-methylbutanedinitrile, butanenitrile, etc.

[0115] "Heterocycle" refers to a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered monocycles, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring of a bicyclic heterocycle can be selected from a saturated ring, an unsaturated ring, and an aromatic ring. Bicyclic heterocycles can be fused, bridged, or spiro ring systems. In some cases, a spiro heterocycle has at least two molecular rings with only one common atom. A spiro heterocycle includes at least one heteroatom.

[0116] "Heterocyclene" refers to a divalent heterocycle that attaches the remainder of the molecule to a radical group.

[0117] "Heteroaryl" or "aromatic heterocycle" refers to a group derived from a heteroaromatic ring group that contains from one to eleven carbon atoms and at least one heteroatom, where each heteroatom can be selected from N, O, and S. As used herein, a heteroaromatic ring can be selected from monocyclic or bicyclic and fused or bridged ring systems, where at least one ring in the ring system is aromatic, i.e., according to Hückel's theory, it contains a cyclic, delocalized (4n + 2)π-electron system. The heteroatoms in a heteroaryl radical can be optionally oxidized. One or more nitrogen atoms (if present) can be optionally quaternized. A heteroaryl can be attached to the remainder of the molecule through any atom of the heteroaryl (where valence allows), such as a carbon atom or a nitrogen atom of the heteroaryl. Examples of heteroaryls include, but are not limited to, pyridine, pyrimidine, oxazole, furan, pyran, thiophene, isoxazole, benzimidazole, benzothiazole, and imidazopyridine.

[0118] "X-membered heteroaryl" is the number of atoms within the ring, i.e., X. For example, a 5-membered heteroaromatic ring or 5-membered aromatic heterocycle has 5 atoms within the ring, such as triazole, oxazole, thiophene, etc.

[0119] The term "unsaturated heterocycle" refers to a heterocycle having at least one degree of unsaturation and not containing an aromatic heterocycle. Examples of unsaturated heterocycles include dihydropyrrole, dihydrofuran, oxazoline, pyrazoline, and dihydropyridine. Heterocycles can be optionally substituted with one or more substituents, such as those described herein.

[0120] The term "substituted" refers to a moiety having a substituent that replaces a hydrogen on one or more carbons or a replaceable heteroatom (e.g., NH of the structure). It should be understood that "substituted" or "substituted with" includes the implicit limitation that such substitution conforms to the allowed valences of the atom being substituted and the substituent and that the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. In certain embodiments, substituted refers to a moiety having a substituent that replaces two hydrogens on the same carbon atom, such as replacing two hydrogens on a single carbon with an oxo, imino, or thio group.

[0121] As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. For suitable organic compounds, the permissible substituents can be one or more and can be the same or different. For the purposes of this disclosure, heteroatoms (such as nitrogen) can have hydrogen substituents and / or any permissible substituents of the organic compounds described herein, which satisfy the valence of the heteroatom. In some embodiments, the substituents can include any of the substituents described herein, for example: halogen, hydroxy, oxo (═O), thioxo (═S), cyano (—CN), nitro (—NO2), imino (═NH), oximino (═N—OH), hydrazino (═N—NH2), -R b -OR a 、-R b -OC(O)-R a 、-R b -OC(O)-OR a 、-R b -OC(O)-N(R a )2、-R b -N(R a )2、-R b -C(O)R a 、-R b -C(O)OR a 、-R b -C(O)N(R a )2、-R b -O-R c -C(O)N((R a )2、-R b -N(R a )C(O)OR a 、-R b -N(R a )C(O)R a 、-R b -N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a)2 (where t is 1 or 2); and alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, and heterocycle, any of which may optionally be substituted with: alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo(=O), thioxo(=S), cyano(-CN), nitro(-NO2), imino(=NH), oximino(=N-OH), hydrazino(=N-NH2), -R b -OR a 、-R b -OC(O)-R a 、-R b -OC(-O)-OR a -、-R b -OC(O)-N(R a )2、-R b -N(R a )2、-R b -C(-O)R a 、-R b -C(O)OR a 、-R b -C(O)N(R a )2、-R b -OR c -C(O)N(R a )2、-R b -N(R a )C(O)OR a 、-R b -N(R a )C(O)R a 、-R b -N(R a )S(O) t R a (where t is 1 or 2)、-R b -S(O) t R a (where t is 1 or 2)、-R b -S(O) t OR a (where t is 1 or 2) and R b -S(O) t N(R a )2 (where t is 1 or 2); where each R a is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, where each R aOptionally, when the valence state permits, it may be substituted by the following: alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo(=O), thioxo(=S), cyano(-CN), nitro(-NO2), imino(=N-H), oximino(=N-OH), hydrazino(=N-NH2), -R b -OR a 、-R b -OC(O)-R a 、-R b -OC(O)-OR a 、-R b -OC(O)-N(R a )2、-R b -N(R a )2、-R b -C(O)R a 、-R b -C(O)OR a 、-R b -C(O)N(R a )2、-R b -O-R c -C(O)N(R a )2、-R b -N(R a )C(O)OR a 、-R b -N(R a )C(O)R a 、-R b -N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a )2(where t is 1 or 2); and where each R b is independently selected from a direct bond or a straight-chain or branched alkylene, alkenylene or alkynylene chain, and each R c is a straight-chain or branched alkylene, alkenylene or alkynylene chain.

[0122] As used in this specification and the claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

[0123] The terms “salt” or “pharmaceutically acceptable salt” refer to salts derived from a variety of organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic bases and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, basic ion exchange resins, and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salts are selected from ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts.

[0124] As used herein, the phrases “parenteral administration” and “administered parenterally” mean modes of administration other than enteral and topical administration, generally by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0125] The phrase “pharmaceutically acceptable” as used herein refers to those compounds, materials, compositions, and / or dosage forms that, within the scope of reasonable medical judgment, are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications and are commensurate with a reasonable benefit / risk ratio.

[0126] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0127] In certain embodiments, the terms "prevent" or "preventing" as related to a disease or disorder can mean that a compound reduces the occurrence of a disorder or condition in a treated sample, relative to an untreated control sample, or delays the onset or reduces the severity of one or more symptoms of a disorder or condition, relative to an untreated control sample.

[0128] As used herein, the term "treat" can include alleviating, reducing, or ameliorating symptoms of a disease or disorder, preventing other symptoms, improving or preventing the underlying cause of symptoms, inhibiting a disease or disorder, e.g., arresting the development of a disease or disorder, alleviating a disease or disorder, causing a disease or disorder to regress, alleviating a condition caused by a disease or disorder, or prophylactically and / or therapeutically halting the symptoms of a disease or disorder.

[0129] The term "ligand" generally refers to a macromolecular compound capable of recognizing and binding to an antigen or receptor associated with a target cell. Ligands can be used to deliver a drug to a population of target cells that bind to the ligand, including but not limited to protein hormones, lectins, growth factors, antibodies, or other substances that can bind to cells, receptors, and / or antigen molecules. A ligand can be an antibody. A ligand can be an antigen-binding fragment. A ligand can be a targeting moiety.

[0130] The term "targeting moiety" refers to a structure that has a selective affinity for a target molecule relative to other non-target molecules. The targeting moiety binds to the target molecule. The targeting moiety can include, for example, an antibody, a peptide, a ligand, a receptor, or a binding portion thereof. The target biomolecule can be a biological receptor or other structure of a cell, such as a tumor antigen.

[0131] The term "linker" refers to a chemical moiety that is capable of combining two different chemical moieties with each other. The linker can include a spacer and an amino acid. The linker can be a cleavable linker that facilitates the release of the compounds described herein. The linker can be used to form a covalent bond with a ligand (such as an antibody).

[0132] The term "antibody" means a whole antibody and any antigen-binding fragment thereof (i.e., "antigen-binding portion") or single-chain variant. A whole antibody is a protein that contains at least two heavy chains (H) and two light chains (L) that are interconnected by disulfide bonds. Each heavy chain contains a heavy-chain variable region (VH) and a heavy-chain constant region that contains three domains: CH1, CH2, and CH3. Each light chain contains a light-chain variable region (VL or Vk) and a light-chain constant region that contains one domain, CL. The VH and VL regions can be further subdivided into hypervariable regions, called complementarity-determining regions (CDRs), which are interspersed with more conserved framework regions (FRs). Each VH and VL contains three CDRs and four FRs, which are arranged in the following order from the amino terminus to the carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable region contains a binding domain that interacts with an antigen. The constant region can mediate the binding of the antibody to host tissues or factors, including various cells of the immune system (such as effector cells) and the first component of the classical complement system (Clq). If the KD of an antibody for binding to antigen X is 5×10 -8 M or lower, more preferably 1×10 -8 M or lower, more preferably 6×10 -9 M or lower, more preferably 3×10 -9 M or lower, even more preferably 2×10 -9 M or lower, then the antibody is said to "specifically bind" to antigen X. The antibody can be a chimeric antibody, a humanized antibody, or preferably a human antibody. The heavy-chain constant region can be engineered to affect the type or degree of glycosylation, extend the half-life of the antibody, enhance or reduce the interaction with effector cells or the complement system, or modulate some other property. The engineering can be accomplished by replacing, adding, or deleting one or more amino acids or by replacing a domain with a domain from another immunoglobulin type or a combination of the foregoing.

[0133] The terms "antigen-binding fragment" and "antigen-binding portion" of an antibody (or simply "antibody portion" or "antibody fragment") mean one or more fragments of an antibody that retain the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody, such as (i) Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragment, a divalent fragment consisting of two Fab fragments linked by a disulfide bridge at the hinge region;

[0134] (iii) Fab' fragment, which is essentially a Fab with a portion of the hinge region (see, e.g., Abbas et al., Cellular and Molecular Immunology, 6th ed., Saunders Elsevier 2007); (iv) Fd fragment consisting of the VH and CH1 domains; (v) Fv fragment consisting of the VL and VH domains of a single arm of an antibody; (vi) dAb fragment (Ward et al., (1989) Nature 341:544 - 546), which consists of the VH domain; (vii) isolated complementarity-determining region (CDR); and (viii) nanobody, a heavy-chain variable region containing a single variable domain and two constant domains. Preferred antigen-binding fragments are Fab, F(ab')2, Fab', Fv, and Fd fragments. In addition, although the two domains VL and VH of the Fv fragment are encoded by different genes, they can be joined by recombinant methods using a synthetic linker so that they can be made into a single protein chain in which the VL and VH regions pair to form a monovalent molecule (referred to as single-chain Fv or scFv); see, e.g., Bird et al. (1988) Science 242:423 - 426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879 - 5883). Such single-chain antibodies are also encompassed within the term "antigen-binding portion" of an antibody.

[0135] The term "isolated antibody" means an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds to antigen X is substantially free of antibodies that specifically bind to antigens other than antigen X). However, an isolated antibody that specifically binds to antigen X may cross-react with other antigens (such as antigen X molecules from other species). In certain embodiments, the isolated antibody specifically binds to a human antigen X and does not cross-react with other (non-human) antigen X. In addition, an isolated antibody may be substantially free of other cellular materials and / or chemicals.

[0136] The term "monoclonal antibody" or "monoclonal antibody composition" means a preparation of antibody molecules consisting of a single molecule that exhibits a single binding specificity and affinity for a particular epitope.

[0137] The term "human antibody" means an antibody having variable regions in which the framework regions and CDR regions (and constant regions, if present) are all derived from human germline immunoglobulin sequences. A human antibody may include post-translational modifications, including natural or synthetic modifications. A human antibody may include amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-directed mutagenesis or by in vivo somatic mutation). However, a "human antibody" does not include an antibody in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto a human framework sequence.

[0138] The term "human monoclonal antibody" means an antibody that exhibits a single binding specificity and has variable regions in which the framework regions and CDR regions are both derived from human germline immunoglobulin sequences. In one embodiment, a human monoclonal antibody is produced by a hybridoma that includes B cells obtained from a transgenic non-human animal (e.g., a transgenic mouse) having a genome that includes a human heavy chain transgene and a light chain transgene that are fused to immortalized cells.

[0139] The compounds of the present disclosure

[0140] The following is a discussion of the compounds and their salts that can be used in the methods of the present disclosure.

[0141] In one aspect, the present disclosure provides compounds of formula (I*):

[0142]

[0143] or a pharmaceutically acceptable salt thereof, wherein;

[0144] L and M are linkers;

[0145] n and u are each independently selected from 0 and 1;

[0146] R 1A is selected from hydrogen and C 1-6 alkyl;

[0147] R 1B is optionally substituted C 1-6 alkyl, wherein the C 1-6 alkyl is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NH2, and oxo;

[0148] wherein when u is 1, R 1B is absent or is -C 1-6 alkylene-O-; and

[0149] wherein either u or n is 1.

[0150] In some embodiments, for the compound or salt of formula (I*), n and u are 1. In some cases, n is 0 and u is 1. In some cases, n is 1 and u is 0. In some cases, n is 0. In some cases, n is 1. In some cases, u is 0. In some cases, u is 1.

[0151] In some embodiments, formula (I*) is represented as

[0152]

[0153] or a pharmaceutically acceptable salt thereof, wherein;

[0154] R 1A is selected from C 1-6 alkyl;

[0155] R 1B is selected from C 1-6 alkyl; and

[0156] L is a linker.

[0157] In some embodiments, formula (I*) or formula (I) is represented as

[0158]

[0159] or a pharmaceutically acceptable salt thereof.

[0160] In some embodiments, for the compound or salt of formula (I*), formula (I), formula (I-A), formula (II) or formula (III), L is L 1 -L 2 -L 3 -L 4 -(L 5 ) m -R 2 ; wherein

[0161] L 1 is selected from C 1-6 alkylene;

[0162] L 2 is selected from optionally substituted C 5-6 carbocycle, wherein C 5-6 carbocycle is optionally substituted with one or more substituents independently selected from R 3 ;

[0163] L 3 is selected from residues comprising 1 to 7 amino acids;

[0164] L 4 is selected from optionally substituted C 1-6 alkylene, wherein C 1-6The alkylene group is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0165] L 5 is selected from (-O-CH2-CH2-) q ;

[0166] R 2 is selected from optionally substituted 5- to 6-membered heterocycles, wherein the 5- to 6-membered heterocycle is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0167] Each R 3 is independently selected from sugars;

[0168] m is selected from 0 and 1; and

[0169] q is selected from 1 to 10.

[0170] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (II), formula (II-A), formula (III) or formula (III-A), R 1A is selected from C 1-6 alkyl and R 1B is selected from C 1-6 alkyl. In some cases, R 1A is the same as R 1B . In some cases, R 1A is different from R 1B . In some cases, R 1A is methyl and R 1B is methyl. In some cases, R 1A is methyl. In some cases, R 1A is hydrogen. In some cases, R 1B is methyl. In some cases, R 1B is C 1-6 alkyl, which is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NH2 and oxo. In some cases, R 1B is C 1-6An alkyl group, which is optionally substituted by one or more substituents independently selected from -OH, -NH2, and oxo. In some cases, R 1B is C 1-6 An alkyl group, which is optionally substituted by one or more substituents independently selected from -OH and oxo. In some cases, R 1B is C alkyl optionally substituted by -OH 1-6 In some cases, R 1B is C alkyl optionally substituted by oxo 1-6 In some cases, R 1B is C 1-6 An alkyl group, which is optionally substituted by one or more substituents independently selected from -NH2 and oxo. In some cases, R 1B is C alkyl optionally substituted by -NH2 1-6 In some cases, R 1B is C alkyl substituted by one -OH 1-6 In some cases, R 1B is

[0171] In some embodiments, for the compound or salt of formula (I*), n is 0, and u is 1, R 1A is selected from hydrogen and C 1-6 alkyl, and R 1B -M is In some cases, R 1B -M is

[0172] In some embodiments, for the compound or salt of formula (I*), formula (I), formula (I-A), formula (II), formula (II-A), formula (III), or formula (III-A), for L, L 1 is selected from C 1-5 alkylene. In some cases, L 1 is selected from C 1-3 alkylene. In some cases, L 1 is C1. In some cases, L 1 is

[0173] In some embodiments, for the compound or salt of formula (I*), formula (I), formula (I-A), formula (II), formula (II-A), formula (III), or formula (III-A), for L, L 2 is selected from optionally substituted C 5-6 carbocycle, wherein the C 5-6 carbocycle is optionally substituted by one or more substituents independently selected from R 3 In some cases, L 2Selected from optionally substituted C 5-6 carbocyclic olefins, wherein C 5-6 carbocyclic olefins are optionally substituted by one or more substituents independently selected from R 3 . In some cases, L 2 is selected from optionally substituted phenyl, wherein phenyl is optionally substituted by one or more substituents independently selected from R 3 . In some cases, L 2 is unsubstituted C 5-6 carbocycle. In some cases, L 2 is phenyl. In some cases, L 2 is phenylene. In some cases, L 2 is In some cases, L 2 is In some cases, L 2 is In some cases, L 2 is In some cases, L 2 is

[0174] In some embodiments, for a compound or salt of formula (I*), formula (I) or formula (I-A), each is represented by formula (I-B):

[0175]

[0176] or a pharmaceutically acceptable salt thereof, wherein;

[0177] b is selected from 0 to 1.

[0178] In some embodiments, for a compound or salt of formula (I*), formula (I-B), b is 1. In some cases, b is 0. In some cases, b is 2.

[0179] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, R 3 is a sugar. In some cases, the sugar is selected from monosaccharides and disaccharides. In some cases, the sugar is selected from monosaccharides. In some cases, the sugar is selected from fructose, galactose, glucose, glucuronic acid, maltose, sucrose and sucrose. In some cases, the sugar is glucuronic acid. In some cases, R 3 is In some cases, R 3 is

[0180] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, L 3 is selected from residues comprising from 1 to 6 amino acids. In some cases, L 3 is selected from residues comprising from 1 to 5 amino acids. In some cases, L 3 is selected from residues comprising from 1 to 4 amino acids. In some cases, L 3 is selected from residues comprising from 1 to 3 amino acids. In some cases, L 3 is selected from residues comprising from 2 to 5 amino acids. In some cases, L 3 is selected from residues comprising from 1 to 2 amino acids. In some cases, L 3 is selected from residues comprising from 2 to 3 amino acids. In some cases, L 3 is selected from residues comprising 1 amino acid. In some cases, L 3 is selected from residues comprising 2 amino acids. In some cases, L 3 is selected from residues comprising 3 amino acids. In some cases, L 3 is selected from residues comprising 4 amino acids. In some cases, L 3 is selected from residues comprising 5 amino acids.

[0181] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, the amino acids are natural amino acids. In some cases, the amino acids are selected from α-amino acids and β-amino acids. In some cases, the amino acids are selected from α-amino acids. In some cases, the amino acids are selected from β-amino acids. In some cases, the amino acids are selected from non-α-amino acids. In some cases, the amino acids are γ-amino acids.

[0182] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, the amino acid is selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, citrulline, and β-alanine. In some cases, the amino acid is selected from alanine, glycine, lysine, phenylalanine, valine, citrulline, and β-alanine. In some cases, the amino acid is selected from alanine, glycine, valine, citrulline, and β-alanine. In some cases, the amino acid is selected from alanine, valine, and citrulline. In some cases, the amino acid is selected from alanine and valine. In some cases, the amino acid is selected from valine and citrulline. In some cases, the amino acid is selected from lysine and phenylalanine. In some cases, the amino acid is selected from glycine and phenylalanine. In some cases, the amino acid is lysine. In some cases, the amino acid is β-alanine.

[0183] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, L 3 is selected from In some cases, L 3 is In some cases, L 3 is In some cases, L 3 is In some cases, L 3 is In some cases, L 3 is In some cases, L 3 is

[0184] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, the amino acid is a non-natural amino acid.

[0185] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, L 4 is selected from C 1-6 alkylene substituted by one oxo. In some cases, L4 Selected from C substituted by one oxo 1-3 alkylene. In some cases, L 4 is selected from In some cases, L 4 is In some cases, L 4 is

[0186] In some embodiments, for the compounds or salts of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, L 5 is selected from (-O-CH2-CH2-) q . In some cases, q is selected from 1 to 5. In some cases, q is selected from 1 to 3. In some cases, q is selected from 1 to 2. In some cases, q is 2. In some cases, q is In some cases, q is

[0187] In some embodiments, for the compounds or salts of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, m is selected from 0 and 1. In some cases, m is 0. In some cases, m is 1.

[0188] In some embodiments, for the compounds or salts of formula (I*), formula (I), formula (I-A) or formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), for L, R 2 is selected from optionally substituted 5-membered heterocycles. In some cases, R 2 is selected from optionally substituted 5-membered heteroalkenes. In some cases, one or more optional substituents of R 2 are selected from oxo and halogen. In some cases, one or more optional substituents of R 2 are selected from oxo. In some cases, the 5-membered heterocycle has at least one double bond. In some cases, the 5-membered heterocycle has one double bond. In some cases, R 2 is maleimide. In some cases, R 2 is In some cases, R 2 is

[0189] In some cases, R 2 is a reactive moiety capable of forming a new bond. In some cases, R 2 is a reactive moiety capable of forming a new bond from an existing double bond.

[0190] In some embodiments, for a compound or salt of formula (I*), n is selected from 0 and 1. In some cases, n is 0. In some cases, n is 1.

[0191] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A) or formula (I-B), L is selected from In some cases, L is selected from In some cases, L is selected from In some cases, L is selected from In some cases, L is selected from

[0192] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A) or formula (I-B), the compound or salt further comprises a ligand. In some cases, the compound or salt is further modified with a ligand. In some cases, the compound or salt is covalently attached to a ligand. In some cases, the compound or salt reacts to form a covalent bond with the ligand. In some cases, the R of the compound or salt 2 reacts to form a covalent bond with the ligand. In some cases, the R of the compound or salt 2 double bond reacts to form a covalent bond with the ligand. In some cases, R 2 is maleimide, and the double bond of maleimide reacts to form a covalent bond with the ligand.

[0193] In some embodiments, a compound or salt of formula (I*), formula (I), formula (I-A) or formula (I-B) is represented by

[0194]

[0195] or a pharmaceutically acceptable salt thereof, wherein;

[0196] Lg is a ligand.

[0197] In some embodiments, formula (II) is represented as

[0198]

[0199] or a pharmaceutically acceptable salt thereof.

[0200] In one aspect, the present disclosure provides a compound of formula (III):

[0201]

[0202] or a pharmaceutically acceptable salt thereof, wherein;

[0203] R 1A is selected from C1-6 alkyl;

[0204] R 1B is an optionally substituted C 1-6 alkyl group, wherein the C 1-6 alkyl group is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NH2, and oxo;

[0205] L is a linker; and

[0206] Lg is a ligand.

[0207] In some embodiments, for a compound or salt of formula (III), the linker (e.g., L) is L 1 -L 2 -L 3 -L 4 -(L 5 ) m -R 2 ; and

[0208] L 1 is selected from C 1-6 alkylene;

[0209] L 2 is selected from an optionally substituted C 5-6 carbocycle, wherein the C 5-6 carbocycle is optionally substituted by one or more substituents independently selected from R 3 ;

[0210] L 3 is selected from residues comprising 1 to 7 amino acids;

[0211] L 4 is selected from an optionally substituted C 1-6 alkylene group, wherein the C 1-6 alkylene group is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0212] L 5 is selected from (-O-CH2-CH2-) q ;

[0213] R 2 is selected from an optionally substituted 5- to 6-membered heterocycle, wherein the 5- to 6-membered heterocycle is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10Halogenoalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 substituted by substituents of alkynyl;

[0214] Each R 3 is independently selected from sugars;

[0215] m is selected from 0 and 1; and

[0216] q is selected from 1 to 10.

[0217] In some embodiments, formula (III) is represented as

[0218]

[0219] or a pharmaceutically acceptable salt thereof.

[0220] In some embodiments, for the compound or salt of formula (I*), only one of u or n is 1. In some cases, u or n is 1 and the other variable is 0.

[0221] In some embodiments, for the compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), m is selected from 0 and 1. In some cases, m is 0. In some cases, m is 1.

[0222] In some embodiments, for the compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), n is selected from 0 and 1. In some cases, n is 0. In some cases, n is 1.

[0223] In some embodiments, for the compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III) or formula (III-A), m is selected from 0 and 1. In some cases, m is 0. In some cases, m is 1.

[0224] In some embodiments, the compound or salt of formula (I*) is represented by:

[0225]

[0226] or a pharmaceutically acceptable salt thereof, wherein;

[0227] R 1A is selected from hydrogen and C 1-6 alkyl;

[0228] R 1B is absent or is -C1-6 Alkylene-O-;

[0229] M is (L 2 ) x -(L 2B ) z -(L 2C ) y -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 ;

[0230] L 2 is selected from C 1-6 alkylene;

[0231] L 2B is selected from (NR 4 ) t C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted by one or more R 5 substituents;

[0232] L 2C is selected from C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted by one or more R 6 substituents;

[0233] L 3 is selected from residues comprising 1 to 7 amino acids;

[0234] L 4 is selected from optionally substituted C 1-6 alkylene, wherein the C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0235] L 5 is selected from (O-CH2-CH2) q -(NR 3 ) s ;

[0236] L 6 is selected from optionally substituted C 1-6 alkylene, wherein the C 1-6The alkylene group is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0237] L 7 is selected from C 5-6 carbocyclic rings;

[0238] R 2 is selected from optionally substituted 5- to 6-membered heterocycles, wherein the 5- to 6-membered heterocycle is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0239] R 3 is selected from hydrogen and C 1-6 alkyl;

[0240] R 4 is selected from hydrogen and C 1-6 alkyl optionally substituted by one or more SO2C 1-6 alkyl;

[0241] Each R 5 is independently selected from sugars;

[0242] Each R 6 is independently selected from halogen, -OH, -CN, -NO2, -NH2, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0243] m is selected from 0 and 1;

[0244] p is selected from 0 and 1;

[0245] q is selected from 0 to 8;

[0246] s is selected from 0 and 1;

[0247] t is selected from 0 and 1;

[0248] x is selected from 0 and 1;

[0249] y is selected from 0 and 1; and

[0250] z is selected from 0 and 1.

[0251] In some embodiments, for a compound or salt of formula (IV), R 1A is selected from hydrogen and C 1-6 alkyl. In some cases, R 1A is hydrogen. In some cases, R 1A is C 1-6 alkyl. In some cases, R 1A is methyl.

[0252] In some embodiments, for a compound or salt of formula (IV), R 1B is absent or is -C 1-6 alkylene-O-. In some cases, R 1B is absent. In some cases, R 1B is -C 1-6 alkylene-O-. In some cases, R 1B is -C2 alkylene-O-.

[0253] In some embodiments, for a compound or salt of formula (IV), R 1B -M is

[0254] In some embodiments, a compound or salt of formula (I*) or formula (IV) is represented by formula (IV-A):

[0255]

[0256] or a pharmaceutically acceptable salt thereof.

[0257] In some embodiments, for a compound or salt of formula (I*), formula (IV), or formula (IV-A), R 1A is hydrogen. In some cases, R 1A is C 1-6 alkyl. In some cases, R 1A is methyl.

[0258] In some embodiments, a compound or salt of formula (I*) or formula (IV) is represented by formula (IV-B):

[0259]

[0260] or a pharmaceutically acceptable salt thereof.

[0261] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), M is L 2 -(L 2B ) z-L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 。In some cases, M is L 2 -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 。In some cases, M is L 2 -L 3 -L 4 -L 5 -L 6 -R 2 。In some cases, M is L 2 -L 3 -L 4 -L 7 -R 2 。In some cases, M is L 2 -L 3 -L 4 -R 2 。In some cases, M is L 2 -L 2B -L 3 -L 4 -L 5 -R 2 。In some cases, M is L 2 -L 2B -L 3 -L 4 -R 2 。In some cases, M is L 2B -L 3 -L 4 -R 2 。

[0262] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), M is (L 2B ) z -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R2 In some cases, M is L 2B -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 In some cases, M is L 2B -L 3 -L 4 -L 5 -L 6 -R 2 In some cases, M is L 2B -L 3 -L 4 -L 5 -R 2 In some cases, M is L 2B -L 3 -L 4 -R 2 。

[0263] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), M is (L 2C ) y -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 In some cases, M is L 2C -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 In some cases, M is L 2C -L 3 -L 4 -L 5 -L 6 -R 2 In some cases, M is L 2C -L 3 -L 4 -L 5 -R 2 In some cases, M is L 2C -L 3 -L 4 -R 2. In some cases, M is L 2C -L 3 -L 4 -L 7 -R 2 . In some cases, L is L 3 -L 4 -R 2 .

[0264] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 2 is selected from C 1-6 alkylene. In some cases, L 2 is C5 alkylene. In some cases, L 2 is C4 alkylene. In some cases, L 2 is C3 alkylene. In some cases, L 2 is C2 alkylene. In some cases, L 2 is C1 alkylene. In some cases, L 2 is selected from C 1-5 alkylene. In some cases, L 2 is selected from C 1-3 alkylene. In some cases, L 2 is C1 alkylene.

[0265] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, z is 1. In some cases, L 2B is selected from NR 4 C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted by one R 5 substituent. In some cases, L 2B is selected from NR 4 C(O)O-CH2-phenylene, wherein the phenylene is optionally substituted by one R 5 substituent. In some cases, L 2B is selected from C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted by one R 5 substituent. In some cases, L 2B is In some cases, L 2B is In some cases, L 2B is In some cases, L 2B is In some cases, L 2B is In some cases, L 2B is In some cases, each R 5 is independently selected from sugars. In some cases, the sugar is selected from monosaccharides and disaccharides. In some cases, the sugar is selected from monosaccharides. In some cases, the sugar is selected from fructose, galactose, glucose, glucuronic acid, maltose, sucrose, and trehalose. In some cases, the sugar is glucuronic acid. In some cases, R 5 is In some cases, R 5 is In some cases, z is 0. In some cases, R 4 is selected from hydrogen. In some cases, R 4 is selected from C 1-6 alkyl optionally substituted by one SO2C 1-6 alkyl. In some cases, R 4 is selected from C 1-6 alkyl substituted by one SO2C 1-6 alkyl. In some cases, R 4 is selected from C 1-6 alkyl substituted by one SO2 methyl. In some cases, R 4 is

[0266] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), for M, y is 1. In some cases, L 2C is selected from C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted by one or more R 6 groups. In some cases, L 2C is selected from C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted by one R 6 group. In some cases, L 2C is In some cases, L 2C is

[0267] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), for M, L 3 is selected from residues comprising from 1 to 7 amino acids. In some cases, L 3 is selected from residues comprising from 1 to 5 amino acids. In some cases, L 3 is selected from residues comprising from 1 to 4 amino acids. In some cases, L 3 is selected from residues comprising from 1 to 3 amino acids. In some cases, L3 Residues selected from those containing 1 to 2 amino acids. In some cases, L 3 Residues selected from those containing 2 to 4 amino acids. In some cases, L 3 Residues selected from those containing 1 amino acid. In some cases, L 3 Residues selected from those containing 2 amino acids. In some cases, L 3 Residues selected from those containing 3 amino acids. In some cases, L 3 Residues selected from those containing 4 amino acids. In some cases, L 3 Residues selected from those containing 5 amino acids. In some cases, L 3 Residues selected from those containing 6 amino acids. In some cases, L 3 Residues selected from those containing 7 amino acids.

[0268] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 3 Selected from residues containing natural and unnatural amino acids.

[0269] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 3 Selected from residues containing natural amino acids. In some cases, L 3 Selected from residues containing α-amino acids. In some cases, L 3 Selected from residues containing β-amino acids.

[0270] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, the amino acid is an unnatural amino acid.

[0271] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 3 Selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, citrulline and β-alanine. In some cases, L 3Selected from alanine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, isoleucine, leucine, lysine, phenylalanine, serine, valine, citrulline, sarcosine, and β-alanine. In some cases, L 3 Selected from alanine, glycine, lysine, phenylalanine, serine, valine, citrulline, sarcosine, and β-alanine. In some cases, L 3 Selected from alanine, glycine, phenylalanine, valine, citrulline, and β-alanine. In some cases, L 3 Selected from alanine, valine, and citrulline. In some cases, L 3 Selected from glycine and phenylalanine. In some cases, L 3 Selected from glycine, phenylalanine, and sarcosine. In some cases, L 3 Selected from sarcosine.

[0272] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), for M, L 3 Comprises 4 amino acids. In some cases, L 3 Comprises 2 amino acids. In some cases, L 3 Comprises 1 amino acid. In some cases, L 3 Comprises at least two different amino acids. In some cases, L 3 is In some cases, L 3 is In some cases, L 3 is In some cases, L 3 is

[0273] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), for M, L 4 Selected from optionally substituted C 1-6 alkylene, wherein C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl. In some cases, L 4 Selected from optionally substituted C 1-6 alkylene, wherein C 1-6The alkylene group is optionally substituted by one or more substituents independently selected from -OH, -NH2, and oxo. In some cases, L 4 is selected from optionally substituted C 1-6 alkylene, wherein the C 1-6 alkylene is optionally substituted by oxo. In some cases, L 4 is selected from -C(O)-(CH2)2- and -C(O)-(CH2)5-. In some cases, L 4 is -C(O)-(CH2)2-. In some cases, L 4 is -C(O)-(CH2)3-. In some cases, L 4 is -C(O)-(CH2)4-. In some cases, L 4 is -C(O)-(CH2)5-.

[0274] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), for M, L 5 is selected from -(O-CH2-CH2-) q -(NR 3 ) s -; wherein, q is selected from 0 to 8; and s is selected from 0 and 1. In some cases, L 5 is selected from -(O-CH2-CH2-) q -(NR 3 ) s -; wherein, q is selected from 0 to 8; and s is 0. In some cases, L 5 is selected from -(O-CH2-CH2-) q -(NR 3 ) s -; wherein, q is selected from 0 to 8; and s is 1. In some cases, q is 1. In some cases, q is 2. In some cases, q is 3. In some cases, q is 4. In some cases, q is 5. In some cases, q is 6. In some cases, L 5 is -(O-CH2-CH2-)2-NH-. In some cases, L 5 is -(O-CH2-CH2-)3-NH-. In some cases, L 5 is -(O-CH2-CH2-)4-NH-. In some cases, L 5 is -(O-CH2-CH2-)5-NH-. In some cases, L 5 is -(O-CH2-CH2-)6-NH-. In some cases, L 5is -(O-CH2-CH2-)7-NH-. In some cases, L 5 is -(O-CH2-CH2-)8-NH-. In some cases, L 5 is selected from -(O-CH2-CH2-) 1-8 . In some cases, s is 0. In some cases, s is 1.

[0275] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 6 is selected from optionally substituted C 1-6 alkylene, wherein C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl. In some cases, L 6 is selected from optionally substituted C 1-6 alkylene, wherein C 1-6 alkylene is optionally substituted by one or more substituents independently selected from -OH, -NH2 and oxo. In some cases, L 6 is selected from optionally substituted C 1-6 alkylene, wherein C 1-6 alkylene is optionally substituted by oxo. In some cases, L 6 is selected from -C(O)-(CH2)2- and -C(O)-(CH2)5-. In some cases, L 6 is -C(O)-(CH2)2-. In some cases, L 6 is -C(O)-(CH2)3-. In some cases, L 6 is -C(O)-(CH2)4-. In some cases, L 6 is -C(O)-(CH2)5-.

[0276] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 7 is selected from C 5-6 carbocycle. In some cases, L 7 is selected from C 5-6 carbocyclene. In some cases, L 7 is selected from C6 carbocycle. In some cases, L 7 is phenyl. In some cases, L 7is phenylene. In some cases, L 7 is cyclohexyl. In some cases, L 7 is In some cases, L 7 is

[0277] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, R 2 is selected from an optionally substituted 5- to 6-membered heterocycle, wherein the 5- to 6-membered heterocycle is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl. In some cases, R 2 is selected from an optionally substituted 5- to 6-membered heteroalkene, wherein the 5- to 6-membered heteroalkene is optionally substituted with one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl. In some cases, R 2 is selected from an optionally substituted 5- to 6-membered heterocycle, wherein the 5- to 6-membered heterocycle is optionally substituted with one or more substituents independently selected from oxo and halogen. In some cases, R 2 is selected from an optionally substituted 5- to 6-membered heteroalkene, wherein the 5- to 6-membered heteroalkene is optionally substituted with one or more substituents independently selected from oxo and halogen. In some cases, R 2 is selected from an optionally substituted 5-membered heterocycle, wherein the 5-membered heterocycle is optionally substituted with one or more substituents independently selected from oxo. In some cases, R 2 is selected from an optionally substituted 5-membered heteroalkene, wherein the 5-membered heteroalkene is optionally substituted with one or more substituents independently selected from oxo. In some cases, the 5-membered heterocycle has at least one double bond. In some cases, the 5-membered heterocycle has one double bond. In some cases, R 2 is maleimide. In some cases, R 2 is In some cases, R 2 is wherein Lg is a ligand. In some cases, R 2 is a reactive moiety capable of forming a new bond. In some cases, R2 is a reactive moiety capable of forming a new bond from an existing double bond.

[0278] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 2 -L 3 -L 4 -R 2 is selected from In some cases, L 2 -L 3 -L 4

[0279]

[0280] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 2 -L 3 -L 4 is In some cases, L 2 -L 3 is

[0281] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 2 -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 is In some cases, L 2 -L 3 -L 4 -L 5 -L 6 -L 7 is In some cases, L 2 -L 3 -L 4 -L 5 -L 6 is

[0282] In some cases, L 2 -L 3 -L 4 -L 5 is In some cases, L 2 -L 3 -L 4 -L 5 is

[0283] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), for M, L 2 -L 2B -L 3 -L 4 -R 2 is In some cases, L 2 -L 2B -L 3 -L 4 is In some cases, L 2 -L 2B -L 3 is In some cases, L 2 -L 2B is In some cases, L 2B is

[0284] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), M is

[0285]

[0286] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), M is selected from L 2C -L 3 -L 4 -R 2 , L 2C -L 3 -L 4 -L 7 -R 2 and L 2C -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 ; wherein L 2C is selected from

[0287]

[0288] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), m is selected from 0 and 1. In some cases, m is 0. In some cases, m is 1.

[0289] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), p is selected from 0 and 1. In some cases, p is 0.

[0290] In some cases, p is 1.

[0291] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), q is selected from 0 and 1. In some cases, q is 0.

[0292] In some cases, q is 1.

[0293] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), s is selected from 0 and 1. In some cases, s is 0.

[0294] In some cases, s is 1.

[0295] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), t is selected from 0 and 1. In some cases, t is 0. In some cases, t is 1.

[0296] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), x is selected from 0 and 1. In some cases, x is 0.

[0297] In some cases, x is 1.

[0298] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C) or formula (IV-D), y is selected from 0 and 1. In some cases, y is 0.

[0299] In some cases, y is 1.

[0300] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), z is selected from 0 and 1. In some cases, z is 0.

[0301] In some cases, z is 1.

[0302] In some embodiments, for a compound or salt of formula (I*), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), the compound or salt further comprises a ligand. In some cases, the compound or salt is further modified with a ligand. In some cases, the compound or salt is covalently attached to a ligand. In some cases, the compound or salt reacts to form a covalent bond with the ligand. In some cases, the R of the compound or salt 2 reacts to form a covalent bond with the ligand. In some cases, the R of the compound or salt 2 of the double bond reacts to form a covalent

[0303] bond with the ligand. In some cases, R 2 is maleimide, and the double bond of maleimide reacts to form a covalent bond with the ligand.

[0304] In some embodiments, for a compound or salt of formula (I*) or formula (I), where u is 0; n is 1; L is L 1 -L 2 -L 3 -L 4 -R 2 ; L 1 is selected from C 1-6 alkylene; L 2 is selected from phenylene; L 3 is selected from residues comprising 1 to 7 amino acids; L 4 is selected from optionally substituted C 1-6 alkylene, where C 1-6 alkylene is substituted with one or more substituents independently selected from oxo; R 1A is methyl, R 1B is methyl, and R 2 is maleimide. In some cases, L 1 is In some cases, L 2 is In some cases, L 3 is selected from residues comprising 1 to 3 amino acids. In some cases, L 3 is selected from residues comprising 2 amino acids. In some cases, L 3 is In some cases, L 4 is selected from In some cases, L 4 is selected from In some cases, R 2 is In some cases, R 2 is wherein Lg is a ligand. In some cases, the ligand is an antibody or an antigen-binding fragment thereof.

[0305] In some embodiments, for a compound or salt of formula (I*) or formula (I), wherein L is

[0306]

[0307] In some embodiments, the compound is represented by formula (VII)

[0308]

[0309] or a pharmaceutically acceptable salt thereof.

[0310] In some embodiments, the compound is represented by formula (VIII)

[0311]

[0312] or a pharmaceutically acceptable salt thereof, wherein DAR is selected from about 1-8, and Lg is a ligand. In some cases, the ligand is a targeting moiety. In some cases, the ligand is an antibody or an antigen-binding fragment thereof. In some embodiments, DAR is about 6. In some embodiments, DAR is about 4. In some embodiments, DAR is about 2. In some cases, DAR is selected from about 4-8.

[0313] In some embodiments, the compound is represented as

[0314]

[0315] or a pharmaceutically acceptable salt thereof, wherein DAR is selected from about 1-8.

[0316] In some embodiments, DAR is about 6. In some embodiments, DAR is about 4.

[0317] In some embodiments, DAR is about 2.

[0318] Included in the present disclosure are salts, particularly pharmaceutically acceptable salts of the compounds described herein. Compounds of the present invention having sufficient acidity, sufficient basicity, or both functional groups can react with a variety of inorganic bases, as well as inorganic and organic acids, to form salts. Alternatively, inherently charged compounds (such as compounds having a quaternary nitrogen) can form salts with appropriate counterions (such as halides such as bromide, chloride, or fluoride, particularly bromide).

[0319] A chemical entity having a carbon-carbon double bond or a carbon-nitrogen double bond can exist in the Z- or E-form (or cis or trans). In addition, some chemical entities can exist in various tautomeric forms. Unless otherwise stated, the compounds described herein are also intended to include all Z-, E-, and tautomeric forms.

[0320] “Tautomers” refer to molecules in which a proton can transfer from one atom of the molecule to another atom of the same molecule. In certain embodiments, the compounds described herein exist as tautomers. Where tautomerism can occur, there will be a chemical equilibrium between the tautomers. The exact ratio of the tautomers depends on several factors, including physical state, temperature, solvent, and pH. Some examples of tautomeric equilibria include:

[0321]

[0322] In some embodiments, the compounds disclosed herein are used in different enriched isotope forms, for example, in 2 H, 3 H, 11 C, 13 C, and / or 14 C enriched content. In a particular embodiment, the compound is deuterated at at least one position. Such deuterated forms can be prepared by the procedures described in U.S. Patent Nos. 5,846,514 and 6,334,997. As described in U.S. Patent Nos. 5,846,514 and 6,334,997, deuteration can improve metabolic stability and / or efficacy, thereby prolonging the duration of action of the drug.

[0323] Unless otherwise stated, the compounds described herein are intended to include compounds that differ only in the presence of one or more isotope-enriched atoms. For example, compounds having this structure but with hydrogen replaced by deuterium or tritium, or carbon replaced by carbon enriched in 13 C or 14 C are all within the scope of the present disclosure.

[0324] The compounds of the present disclosure optionally contain non-natural proportions of atomic isotopes at one or more atoms that make up such compounds. For example, the compounds can be labeled with isotopes, such as deuterium ( 2 H), tritium (3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C). Isotopic substitution with 2 H, 11 C, 13 C, 14 C, 15 C, 12 N, 13 N, 15 N, 16 N, 16 O, 17 O, 14 F, 15 F, 16 F, 17 F, 18 F, 33 S, 34 S, 35 S, 36 S, 35 Cl, 37 Cl, 79 Br, 81 Br, and 125 I is also contemplated. All isotopic variants of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

[0325] In certain embodiments, some or all of the 1 H of the compounds disclosed herein is replaced by 2 H. Methods for synthesizing deuterium-containing compounds are known in the art and include, but are not limited to, the following synthetic methods.

[0326] Deuterium-substituted compounds are synthesized using various methods described in the following literature: Dean, Dennis C.; editor. Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr., Pharm. Des., 2000; 6(10)] 2000, 110 pp; George W.; Varma, Rajender S. The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans, E. Anthony. Synthesis of Radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.

[0327] Deuterated starting materials are readily available and are used in the synthetic methods described herein for the synthesis of deuterium-containing compounds. A large number of deuterated reagents and building blocks are available from chemical suppliers such as Aldrich Chemical Co.

[0328] The compounds of the present invention also include crystalline and amorphous forms of these compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, non-solvated polymorphs (including anhydrous compounds), conformational polymorphs, and amorphous forms of the compounds, and mixtures thereof.

[0329] The compounds described herein can, in some cases, exist as diastereoisomers, enantiomers or other stereoisomeric forms. If no absolute stereochemistry is specified, the compounds described herein include all diastereoisomers, enantiomers and epimeric forms and their appropriate mixtures. Separation of stereoisomers can be carried out by chromatography or by formation of diastereoisomers and by recrystallization or chromatography or any combination thereof. (Jean Jacques, Andre Collet, Samuel H. Wilen, “Enantiomers, Racemates and Resolutions”, John Wiley And Sons, Inc., 1981, which is incorporated herein by reference for that disclosure). Stereoisomers can also be obtained by stereoselective synthesis.

[0330] The methods and compositions described herein include the use of amorphous forms as well as crystalline forms (also known as polymorphs). The compounds described herein can be in the form of pharmaceutically acceptable salts. In addition, in some embodiments, the active metabolites of these compounds having the same type of activity are included within the scope of the present disclosure. Further, the compounds described herein can exist in unsolvated form or in solvated form with a pharmaceutically acceptable solvent such as water, ethanol, etc. The solvated forms of the compounds described herein are also considered to be disclosed herein.

[0331] In certain embodiments, a compound or a salt of a compound can be a prodrug, for example, where a hydroxyl group in the parent compound is present as an ester or a carbonate or a carboxylic acid in the parent compound. The term “prodrug” is intended to cover compounds that are converted into the agents of the present disclosure under physiological conditions. One method for preparing a prodrug is to include hydrolysis of one or more selected moieties under physiological conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by the enzymatic activity of a host animal such as a particular target cell within the host animal. For example, esters or carbonates (e.g., esters of alcohols or carboxylic acids and carbonates and esters of phosphonic acids) are preferred prodrugs of the present disclosure.

[0332] Prodrug forms of the compounds described herein, where the prodrug is metabolized in vivo to produce the compounds as shown herein, are included within the scope of the claims. In some cases, some of the compounds described herein can be prodrugs of another derivative or active compound.

[0333] Prodrugs are generally useful because in some cases they may be more readily administered than the parent drug. For example, they may be bioavailable by oral administration while the parent is not. Prodrugs can help enhance the cellular permeability of a compound relative to the parent drug. The solubility of a prodrug in a pharmaceutical composition may also be higher than that of the parent drug. Prodrugs can be designed as reversible drug derivatives to act as modifiers to enhance the transport of a drug to a site-specific tissue or increase the residence of a drug within a cell.

[0334] In some embodiments, the prodrug is designed to increase the lipophilicity of the agent. In some embodiments, the prodrug is designed to increase effective water solubility. See, e.g., Fedorak et al., Am. J. Physiol., 269:G210-218 (1995); McLoed et al., Gastroenterol, 106:405-413 (1994); Hochhaus et al., Biomed. Chrom., 6:283-286 (1992); J. Larsen and H. Bundgaard, Int. J. Pharmaceutics, 37, 87 (1987); J. Larsen et al., Int. J. Pharmaceutics, 47, 103 (1988); Sinkula et al., J. Pharm. Sci., 64:181-210 (1975); T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series; and Edward B. Roche, Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, all of which are incorporated herein by reference for such disclosure. According to another embodiment, the present disclosure provides methods for producing the compounds defined above. The compounds can be synthesized using conventional techniques. Advantageously, these compounds are conveniently synthesized from readily available starting materials.

[0335] Synthetic chemical transformations and methods that can be used to synthesize the compounds described herein are known in the art, including, for example, those described in R. Larock, Comprehensive Organic

[0336] Those described in Transformations (1989); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d. Ed. (1991); L. Fieser and M. Fieser, Fieser and Fieser’s Reagents for Organic Synthesis (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis (1995).

[0337] ligand

[0338] In some embodiments, for a compound or salt of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III), formula (III-A), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), formula (IV-D), or formula (VIII), the ligand is selected from an antibody or an antigen-binding fragment thereof.

[0339] In some cases, the ligand is selected from chimeric antibodies, humanized antibodies, and human antibodies.

[0340] In some embodiments, the ligand (e.g., an antibody) performs a targeting function. By binding to the target tissue or cell where its antigen or receptor is located, the ligand directs the conjugate thereto. In some cases, when the ligand is an antibody, the compound is sometimes referred to as an antibody-drug conjugate (ADC) or an immunoconjugate. Preferably, the target tissue or cell is a cancer tissue or cell, and the antigen or receptor is a tumor-associated antigen, i.e., an antigen that is uniquely expressed or overexpressed by cancer cells as compared to non-cancerous cells. In some cases, the conjugate is internalized into the target cell by endocytosis and cleavage occurs within the target cell.

[0341] In some embodiments, the ligand is an antibody against a tumor-associated antigen, allowing for selective targeting of cancer cells. Examples of such antigens include: mesothelin, prostate-specific membrane antigen (PSMA), CD19, CD22, CD30, CD70, B7H3, B7H4 (also known as O8E), protein tyrosine kinase 7 (PTK7), glypican-3, RG1, fucosyl-GM1, CTLA-4, and CD44. The antibody can be animal (e.g., murine), chimeric, humanized, or preferably human. The antibody is preferably a monoclonal antibody, especially a monoclonal human antibody. The preparation of monoclonal human antibodies against some of the above antigens is disclosed in the following: Korman et al., U.S. Patent No. 8,609,816B2 (2013; B7H4, also known as O8E; especially antibodies 2A7, 1G11, and 2F9); Rao-Naik et al., U.S. Patent No. 8,097,703B2 (2012; CD19; especially antibodies 5G7, 13F1, 46E8, 21D4, 21D4a, 47G4, 27F3, and 3C10); King et al., U.S. Patent No. 8,481,683B2 (2013; CD22; especially antibodies 12C5, 19A3, 16F7, and 23C6); Keler et al., U.S. Patent No. 7,387,776B2 (2008; CD30; especially antibodies 5F11, 2H9, and 17G1); Terrett et al., U.S. Patent No. 8,124,738B2 (2012; CD70; especially antibodies 2H5, 10B4, 8B5, 18E7, and 69A7); Korman et al., U.S. Patent No. 6,984,720B1 (2006; CTLA-4; especially antibodies 10D1, 4B6, and 1E2); Vistica et al., U.S. Patent No. 8,383,118B2 (2013, fucosyl-GM1, especially antibodies 5B1, 5B1a, 7D4, 7E4, 13B8, and 18D5); Korman et al., U.S. Patent No. 8,008,449B2 (2011; PD-1; especially antibodies 17D8, 2D3, 4H1, 5C4, 4A11, 7D3, and 5F4); Huang et al., US2009 / 0297438A1 (2009; PSMA; especially antibodies 1C3, 2A10, 2F5, 2C6); Cardarelli et al., U.S. Patent No. 7,875,278B2 (2011; PSMA; especially antibodies 4A3, 7F12, 8C12, 8A11, 16F9, 2A10, 2C6, 2F5, and 1C3); Terrett et al., U.S. Patent No. 8,222,375B2 (2012; PTK7; especially antibodies 3G8, 4D5, 12C6, 12C6a, and 7C8);Terrett et al., U.S. Patent No. 8,680,247 B2 (2014; phosphatidylinositol glycan-3; particularly antibodies 4A6, 11E7, and 16D10); Harkins et al., U.S. Patent No. 7,335,748 B2 (2008; RG1; particularly antibodies A, B, C, and D); Terrett et al., U.S. Patent No. 8,268,970 B2 (2012; mesothelin; particularly antibodies 3C10, 6A4, and 7B1); Xu et al., US2010 / 0092484 A1 (2010; CD44; particularly antibodies 14G9.B8.B4, 2D1.A3.D12, and 1A9.A6.B9); Deshpande et al., U.S. Patent No. 8,258,266 B2 (2012; IP10; particularly antibodies 1D4, 1E1, 2G1, 3C4, 6A5, 6A8, 7C10, 8F6, 10A12, 10A2S, and 13C4); Kuhne et al., U.S. Patent No. 8,450,464 B2 (2013; CXCR4; particularly antibodies F7, F9, D1, and E2); and Korman et al., U.S. Patent No. 7,943,743 B2 (2011; PD-L1; particularly antibodies 3G10, 12A4, 10A5, 5F8, 10H10, 1B12, 7H1, 11E6, 12B7, and 13G4); the disclosures of which are incorporated herein by reference.;

[0342] In some embodiments, the ligand can also be an antibody fragment or an antibody mimetic, such as an affibody, a domain antibody (dAb), a nanobody, a unibody, a DARPin, an anticalin, a versabody, a duocalin, a lipocalin, or an avimer.

[0343] In some embodiments, any of several different reactive groups on the ligand can serve as a conjugation site, including the ε-amino group in a lysine residue, a pendant carbohydrate moiety, a carboxylic acid group, a disulfide group, and a thiol group. Each type of reactive group represents a trade-off, having some advantages and some disadvantages. For a review of antibody reactive groups suitable for conjugation, see, e.g., Garnett, Adv. Drug Delivery Rev. 53 (2001), 171-216 and Dubowchik and Walker, Pharmacology & The Rapeutics 83 (1999), 67-123, the disclosures of which are incorporated herein by reference.

[0344] In some embodiments, the ligand is conjugated via the lysine ε-amino group. Most antibodies have multiple lysine ε-amino groups, which can be conjugated via amide, urea, thiourea, or carbamate bonds using techniques known in the art. However, it is difficult to control which ε-amino groups and how many ε-amino groups react, resulting in potential batch-to-batch variability in conjugate preparation. In addition, conjugation can result in the neutralization of protonated ε-amino groups, which are important for maintaining the native conformation of the antibody, or conjugation can occur at lysines near or at the antigen-binding site, neither of which is an ideal situation.

[0345] In some embodiments, since many antibodies are glycosylated, the ligand can be conjugated via carbohydrate side chains. The carbohydrate side chains can be oxidized with periodate to generate aldehyde groups, which can then react with amines to form imine groups, such as in semicarbazones, oximes, or hydrazones. If desired, the imine group can be converted to a more stable amine group by reduction with sodium cyanoborohydride. For additional disclosure regarding conjugation via carbohydrate side chains, see, for example, Rodwell et al., Proc. Nat'l Acad. Sci. USA 83, 2632-2636 (1986); the disclosure of which is incorporated herein by reference. As with the lysine ε-amino group, there are concerns regarding the reproducibility of the conjugation site location and stoichiometry.

[0346] In some embodiments, the ligand can be conjugated via a carboxylic acid group. In some cases, the terminal carboxylic acid group is functionalized to generate a carbohydrazide, which then reacts with an aldehyde-containing conjugation moiety. See Fisch et al., Bioconjugate Chemistry 1992, 3, 147-153.

[0347] In some embodiments, antibodies can be conjugated by bridging cysteine residues on the antibody with sulfur on the conjugate or another moiety of the compound via a disulfide bond. Some antibodies lack free thiol (mercapto) groups but have disulfide bonds, such as in the hinge region. In such cases, free thiol groups can be generated by reducing the native disulfide bonds. The thiol groups thus generated can then be used for conjugation. See, for example, Packard et al., Biochemistry 1986, 25, 3548 - 3552; King et al., Cancer Res. 54, 6176 - 6185 (1994); and Doronina et al., Nature Biotechnol. 21(7), 778 - 784 (2003); the disclosures of which are incorporated herein by reference. There are various known methods for introducing free thiol groups into antibodies without disrupting native disulfide bonds, and these methods can be practiced with the ligands of the present invention. Depending on the method employed, it may be possible to introduce a predictable number of free thiol groups at a predetermined position. In one method, mutant antibodies are prepared in which cysteine is replaced by another amino acid. See, for example, Eigenbrot et al., U.S. Patent No. 7,521,541 B2 (2009); Chilkoti et al., Bioconjugate Chem. 1994, 5, 504 - 507; Urnovitz et al., U.S. Patent No. 4,698,420 (1987); Stimmel et al., J. Biol. Chem., 275(39), 30445 - 30450 (2000); Bam et al., U.S. Patent No. 7,311,902 B2 (2007); Kuan et al., J. Biol. Chem., 269(10), 7610 - 7618 (1994); Poon et al., J. Biol. Chem., 270(15), 8571 - 8577 (1995). In another method, an additional cysteine is added to the C-terminus. See, for example, Cumber et al., J. Immunol., 149, 120 - 126 (1992); King et al., Cancer Res., 54, 6176 - 6185 (1994); Li et al., Bioconjugate Chem., 13, 985 - 995 (2002); Yang et al., Protein Engineering, 16, 761 - 770 (2003); and Olafson et al., Protein Engineering Design & Selection, 17, 21 - 27 (2004). A preferred method for introducing free cysteine is proposed by Liu et al. in WO2009 / 026274 A1, in which an amino acid sequence bearing cysteine is added to the C-terminus of the antibody heavy chain.This method introduces a known number of cysteine residues (one per heavy chain) at a known position distant from the antigen-binding site. The disclosures of the documents cited in this paragraph are hereby incorporated by reference in their entirety.

[0348] In some embodiments, the lysine ε-amino group can be modified with a reagent such as 2-iminothiolane or N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) to convert the ε-amino group to a thiol or disulfide group, generating a cysteine surrogate, so to speak.

[0349] Linker

[0350] Compounds and salts of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III), formula (III-A), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), formula (IV-D), formula (V), formula (VI), formula (VII) can bind to a linker (e.g., L). The linker can be as described elsewhere herein. In some cases, L is a peptide linker. In some embodiments, the linker also binds to a ligand (e.g., an antibody) and is referred to as an antibody-drug conjugate or conjugate. The linker of the conjugates described herein can not affect the binding of the active portion of the conjugate (e.g., antigen-binding domain, Fc domain, target-binding domain, antibody, agonist, etc.) to the target (which can be a cognate binding partner such as an antigen). The conjugate can contain multiple linkers, each linker having one or more attached compounds. These linkers can be the same linker or different linkers.

[0351] In some embodiments, the linker can be short, flexible, rigid, cleavable, non-cleavable, hydrophilic or hydrophobic. The linker can contain segments with different properties, such as flexible segments or rigid segments. The linker can be chemically stable in the extracellular environment, e.g., chemically stable in the bloodstream, or can include labile or selectively stable linkages. The linker can include linkages that are designed to be cleaved and / or sacrificed or otherwise degraded specifically or non-specifically intracellularly. Cleavable linkers can be sensitive to enzymes. Cleavable linkers can be cleaved by enzymes such as proteases. Cleavable linkers can contain valine-citrulline linkers or valine-alanine peptides. Linkers containing valine-citrulline or valine-alanine can contain maleimide or succinimide groups.

[0352] In some embodiments, the non-cleavable linker can be protease-insensitive. The non-cleavable linker can be a maleimidocaproyl linker. The maleimidocaproyl linker can comprise N-maleimidomethylcyclohexane-1-carboxylate. The maleimidocaproyl linker can contain a succinimide group. The maleimidocaproyl linker can contain a pentafluorophenyl group. The linker can be a combination of a maleimidocaproyl group and one or more polyethylene glycol molecules. The linker can be a maleimide-PEG4 linker. The linker can be a combination of a maleimidocaproyl linker containing a succinimide group and one or more polyethylene glycol molecules. The linker can be a combination of a maleimidocaproyl linker containing a pentafluorophenyl group and one or more polyethylene glycol molecules. The linker can contain a maleimide attached to a polyethylene glycol molecule, where the polyethylene glycol can allow for greater linker flexibility or can be used to extend the linker. The linker can be a (maleimidocaproyl)-(valine-citrulline)-(p-aminobenzyloxycarbonyl) linker. The linker can be a linker suitable for attachment to an engineered cysteine (THIOMAB). The THIOMAB linker can be a (maleimidocaproyl)-(valine-citrulline)-(p-aminobenzyloxycarbonyl) linker.

[0353] In some embodiments, the linker can further comprise alkylene, alkenylene, alkynylene, polyether, polyester, polyamide groups, and further comprise polyamino acids, polypeptides, cleavable peptides or aminobenzyl carbamate. The linker can contain a maleimide at one end and an N-hydroxysuccinimide ester at the other end. The linker can contain N-terminal amine acetylated lysine and a valine-citrulline cleavage site. The linker can be a linkage produced by microbial transglutaminase, where the linkage can be established between an amine-containing moiety and a moiety engineered to contain glutamine, due to the enzyme-catalyzed formation of a bond between the acyl group of the glutamine side chain and the primary amine of the lysine chain. The linker can contain a reactive primary amine. The linker can be a sortase A linker. The sortase A linker can be created by a sortase A enzyme that fuses an LXPTG recognition motif with an N-terminal GGG motif, thereby regenerating a native amide bond. Thus, the created linker can connect the portion attached to the LXPTG recognition motif with the portion attached to the N-terminal GGG motif.

[0354] In some embodiments, any compound or salt described herein is linked to an antibody construct via a linker (also referred to herein as L or linker). As used herein, L can be selected from any of the linker moieties discussed herein. The linker that connects the compound or salt to the conjugate antibody construct can be short, long, hydrophobic, hydrophilic, flexible, or rigid, or can consist of segments that independently have one or more of the above properties such that the linker can include segments with different properties. The linker can be multivalent such that they covalently link more than one compound or salt to a single site on the antibody construct, or can be monovalent such that they covalently link a single compound or salt to a single site on the antibody construct.

[0355] In some embodiments, the linker can have from about 10 to about 500 atoms in the linker, such as from about 10 to about 400 atoms, such as from about 10 to about 300 atoms in the linker. In some embodiments, the linker can have from about 30 to about 400 atoms, such as from about 30 to about 300 atoms in the linker.

[0356] In some embodiments, the linker described herein can link any one of the compounds or salts of the formulas described herein to a ligand (e.g., an antibody) via a covalent linkage between the linker, the antibody construct, and the compound. The linker can include functional groups capable of covalently linking to a ligand (e.g., an antibody).

[0357] In some embodiments, by way of example and not limitation, some cleavable and non-cleavable linkers that can be included in the conjugates described herein are described below.

[0358] In some embodiments, the cleavable linker can be cleaved in vitro and in vivo. The cleavable linker can include chemically or enzymatically labile or degradable linkages. The cleavable linker can rely on intracellular processes to release the benzazepine compound, such as reduction in the cytoplasm, exposure to acidic conditions in lysosomes, or cleavage by specific proteases or other enzymes within the cell. The cleavable linker can contain one or more chemically or enzymatically cleavable chemical bonds while the remainder of the linker can be non-cleavable.

[0359] In some embodiments, the linker can contain chemically labile groups such as hydrazones and / or disulfides. Linkers containing chemically labile groups can take advantage of the differential properties between plasma and some cytoplasmic compartments. Cleavable linkers can also include disulfides.

[0360] In some embodiments, acid-labile groups (such as hydrazones) can remain intact in the systemic circulation at the neutral pH of blood (pH 7.3 - 7.5) and once the antibody construct benzazepine Compound conjugates are internalized into the weakly acidic endosomal (pH 5.0 - 6.5) and lysosomal (pH 4.5 - 5.0) compartments of cells and can undergo hydrolysis and release benzazepines compounds. This pH-dependent release mechanism can be associated with non-specific release of the drug. To increase the stability of the linker hydrazone group, the linker can be modified by chemical modification (e.g., substitution), allowing for adjustment to achieve more efficient release in lysosomes while minimizing losses in circulation. Hydrazone-containing linkers can contain additional cleavage sites, such as additional acid-labile cleavage sites and / or enzymatically labile cleavage sites. Other acid-labile groups that can be included in the linker include cis-aconityl-containing linkers. Cis-aconityl chemistry can use carboxylic acids juxtaposed to the amide bond to accelerate amide hydrolysis under acidic conditions.

[0361] In some embodiments, the linker can be specifically cleaved by an enzyme. For example, the linker can be cleaved by lysosomal enzymes. Such linkers can be peptide-based or can include peptide regions that can serve as substrates for enzymes. Peptide-based linkers can be more stable in the plasma and extracellular environment compared to chemically labile linkers. Peptide bonds can have good serum stability because lysosomal proteolytic enzymes can have very low activity in blood due to the presence of endogenous inhibitors and the inappropriately high pH of blood compared to lysosomes. Release of the compounds described herein from antibody-drug conjugates can occur due to the action of lysosomal proteases (e.g., cathepsins and plasmin). These proteases can be present at elevated levels in certain tumor tissues. The linker can be cleaved by lysosomal enzymes. Lysosomal enzymes can be, for example, cathepsin B, cathepsin S, β-glucuronidase, or β-galactosidase. Cleavable peptides can be selected from tetrapeptides, such as Gly-Phe-Leu-Gly, Ala-Leu-Ala-Leu, or dipeptides, such as Val-Cit, Val-Ala, and Phe-Lys. Dipeptides can have lower hydrophobicity compared to long peptides. A variety of dipeptide-based cleavable linkers can be used in the antibody-drug conjugates described herein. Enzymatically cleavable linkers can be β-glucuronide-based linkers.

[0362] In some embodiments, the cleavable linker can include non-cleavable moieties or segments, and / or cleavable segments or moieties can be included in an otherwise non-cleavable linker to make it cleavable.

[0363] In some embodiments, the linker can contain an enzymatically cleavable peptide moiety. The peptide can be selected from natural amino acids, unnatural amino acids, or combinations thereof. In certain embodiments, the peptide can be selected from tripeptides or dipeptides. In specific embodiments, the dipeptide can comprise L-amino acids and is selected from: Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe-Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu-Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit or salts thereof.

[0364] Pharmaceutical formulation

[0365] In certain embodiments, provided herein are pharmaceutical compositions comprising a therapeutically effective amount of any compound or salt of any one of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III), formula (III-A), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), formula (IV-D), formula (V), formula (VI), formula (VII), formula (VIII) (also referred to herein as "agent").

[0366] The pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers (including excipients and auxiliaries) which assist in processing the medicament into a pharmaceutically useful preparation. Appropriate formulations depend on the chosen route of administration. An overview of pharmaceutical compositions is found, for example, in Remington: The Science and Practice of Pharmacy, 19th Edition (Easton, Pa., Mack Publishing Company, 1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition (Lippincott Williams & Wilkins, 1999).

[0367] The compositions and methods of the present disclosure can be used to treat an individual in need. In certain embodiments, the individual is a mammal such as a human or non-human mammal. When administered to an animal (such as a human), the composition or medicament is preferably administered in the form of a pharmaceutical composition which comprises, for example, the medicament and a pharmaceutically acceptable carrier or excipient. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions (such as water or physiological buffer saline) or other solvents or vehicles (such as ethylene glycol, glycerol, oils (such as olive oil) or injectable organic esters). In a preferred embodiment, when such pharmaceutical compositions are used for human administration, particularly for invasive routes of administration (such as injection or implantation routes which circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free or substantially pyrogen-free. For example, excipients can be selected to achieve delayed release of the agent or to selectively target one or more cells, tissues or organs. The pharmaceutical compositions can be in dosage unit form, such as tablets, capsules, granules, reconstitutable lyophilized agents, powders, solutions, syrups, suppositories, injections, etc. The composition can also be present in a transdermal administration system, such as a skin patch. The composition can also be present in a solution suitable for topical administration, such as eye drops.

[0368] Pharmaceutically acceptable excipients may contain physiologically acceptable agents, such as to stabilize, increase solubility or increase absorption of a compound (such as a pharmaceutical agent). Such physiologically acceptable agents include, for example, carbohydrates (such as glucose, sucrose or dextran), antioxidants (such as ascorbic acid or glutathione), chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of pharmaceutically acceptable excipients (including physiologically acceptable agents) depends, for example, on the route of administration of the composition. The formulation or pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) may also be a liposome or other polymeric matrix, which may have incorporated therein, for example, a compound of the present invention. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable and metabolizable carriers, and their preparation and administration are relatively simple.

[0369] The pharmaceutical composition (formulation) may be administered to a subject by any of a variety of routes of administration, including, for example, oral, such as by drenches in the form of: aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue; absorption through the oral mucosa, such as sublingual absorption; absorption through the anus, rectum or vagina, such as in the form of vaginal suppositories, creams or foams; parenteral administration, including intramuscular, intravenous, subcutaneous or intrathecal, such as in the form of a sterile solution or suspension; nasal administration; intraperitoneal administration; subcutaneous administration; transdermal administration, such as in the form of a patch applied to the skin; and topical administration, such as in the form of a cream, ointment or spray applied to the skin, or in the form of eye drops. The compound may also be formulated for inhalation. In certain embodiments, the compound may simply be dissolved or suspended in sterile water.

[0370] The pharmaceutical composition may be a sterile aqueous or non-aqueous solution, suspension or emulsion, such as a microemulsion. The excipients described herein are only examples and do not constitute any limitation. An effective amount or a therapeutically effective amount refers to the amount of one or more pharmaceutical agents administered to a subject in a single dose or as part of a series of doses, which amount is effective for producing the desired therapeutic effect.

[0371] In general, the therapeutic effect of a subject can be monitored using assays and methods suitable for the condition being treated, which assays are familiar to those skilled in the art and are described herein. The pharmacokinetics of a pharmaceutical agent or one or more of its metabolites administered to a subject can be monitored by determining the level of the pharmaceutical agent or metabolite in a biological fluid from the subject (such as blood, blood fractions (such as serum) and / or urine, and / or other biological samples or biological tissues from the subject). Any method for detecting an agent practiced in the art and described herein can be used to measure the level of the pharmaceutical agent or metabolite during a course of treatment.

[0372] The dosage of the agents described herein for treating a disease or disorder can depend on the condition of the subject, i.e., the stage of the disease, the severity of the symptoms caused by the disease, the overall health status, as well as age, sex, and weight, and other factors that will be apparent to those skilled in the medical arts. The pharmaceutical composition can be administered in a manner suitable for the disease to be treated as determined by those skilled in the medical arts. In addition to the factors related to the use of the agents for treating a disease or disorder described herein and above, the appropriate duration and frequency of administration of the agents can also be determined or adjusted according to factors such as the patient's condition, the type and severity of the patient's disease, the specific form of the active ingredient, and the method of administration. The optimal dosage of the agent can generally be determined using experimental models and / or clinical trials. The optimal dosage can depend on the body mass, weight, or blood volume of the subject. It is generally preferred to use the minimum dosage sufficient to provide effective treatment. The design and conduct of preclinical and clinical studies of the agents described herein (including drugs administered for prophylactic benefits) are entirely within the skills of those skilled in the relevant art. When two or more agents are administered to treat a disease or disorder, the optimal dosage of each agent can be different, such as lower than the dosage when either agent is used alone as a monotherapy. In certain specific embodiments, the combination of two agents can act synergistically or additively, and the amount of either agent can be lower than the amount when administered alone. The amount of the agent that can be administered daily can be, for example, about 0.01 mg / kg to 100 mg / kg, such as about 0.1 to 1 mg / kg, about 1 to 10 mg / kg, about 10 - 50 mg / kg, about 50 - 100 mg / kg body weight. In other embodiments, the amount of the agent that can be administered daily is about 0.01 mg / kg to 1000 mg / kg body weight, about 100 - 500 mg / kg body weight, or about 500 - 1000 mg / kg body weight. The optimal dosage per day or per course of treatment can vary depending on the disease or disorder to be treated and can also vary depending on the route of administration and the treatment regimen.

[0373] The pharmaceutical composition comprising the agent can be formulated in a manner suitable for the delivery method using techniques conventional in the art. The composition can be in solid (e.g., tablets, capsules), semi-solid (e.g., gels), liquid, or gaseous (e.g., aerosols) form. In other embodiments, the pharmaceutical composition is administered by bolus infusion.

[0374] In the pharmaceutical field, pharmaceutically acceptable excipients are well known and described, for example, in Rowe et al., Handbook of Pharmaceutical Excipients: A Comprehensive Guide to Uses, Properties, and Safety, 5th Edition, 2006 and Remington: The Science and Practice of Pharmacy (Gennaro, 21st Edition Mack Pub. Co., Easton, PA (2005)). Exemplary pharmaceutically acceptable excipients include sterile saline and phosphate buffered saline at physiological pH. Preservatives, stabilizers, dyes, buffers, etc. can be provided in the pharmaceutical composition. In addition, antioxidants and suspending agents can also be used. Generally, the choice of excipient type is based on the mode of administration and the chemical composition of the active ingredient. Alternatively, the compositions described herein can be formulated as lyophilized products. The compositions described herein can be lyophilized or otherwise formulated into a lyophilized product using one or more suitable excipient solutions for dissolving and / or diluting the pharmaceutical composition upon administration. In other embodiments, the pharmaceutical agent can be encapsulated in liposomes using techniques known and practiced in the art. In certain specific embodiments, the pharmaceutical agent is not formulated in liposomes for use in stents for treating highly (but not completely) occluded arteries. The pharmaceutical composition can be formulated for any suitable mode of administration described herein and in the art.

[0375] Pharmaceutical compositions, such as those for oral administration or injection, infusion, subcutaneous delivery, intramuscular delivery, intraperitoneal delivery, or other methods, can be in liquid form. Liquid pharmaceutical compositions can include, for example, one or more of the following: sterile diluents such as water, saline solutions (preferably physiological saline), Ringer's solution, isotonic sodium chloride, fixed oils that can be used as solvents or suspending media, polyethylene glycol, glycerin, propylene glycol, or other solvents; antibacterial agents; antioxidants; chelating agents; buffers, and agents for adjusting tonicity such as sodium chloride or dextrose. Parenteral compositions can be enclosed in ampoules, disposable syringes, or multi-dose vials made of glass or plastic. Physiological saline is preferably used, and injectable pharmaceutical compositions are preferably sterile. In another embodiment, for treating ophthalmic conditions or diseases, the liquid pharmaceutical composition can be administered to the eye in the form of eye drops. The liquid pharmaceutical composition can be delivered orally.

[0376] For oral formulations, at least one of the agents described herein can be used alone or in combination with suitable additives to form tablets, powders, granules or capsules, and used in combination with diluents, buffers, wetting agents, preservatives, colorants and flavoring agents, if desired. The agent can be formulated with a buffer to provide protection of the compound from the low pH of the gastric environment and / or an enteric coating. The agent included in the pharmaceutical composition can be formulated with a flavoring agent (e.g., in liquid, solid or semi-solid formulations) and / or with an enteric coating for oral administration.

[0377] A pharmaceutical composition comprising any one of the agents described herein can be formulated for sustained or slow release, also known as timed release or controlled release. Such compositions can generally be prepared using well-known techniques and administered, for example, orally, rectally, intradermally or subcutaneously by implantation, or implanted at the desired target site. Sustained release formulations can contain a compound dispersed in a carrier matrix and / or a compound contained in a reservoir surrounded by a rate-controlling membrane. The excipients used in such formulations are biocompatible and can also be biodegradable; preferably the formulation provides a relatively constant level of release of the active ingredient. The amount of the agent contained in the sustained release formulation depends on the implantation site, the release rate and the expected duration, as well as the nature of the condition, disease or disorder to be treated or prevented.

[0378] In certain embodiments, a pharmaceutical composition comprising an agent is formulated for transdermal, intradermal or topical administration. The composition can be administered in the form of a powder / talc or other solid, liquid, spray, aerosol, ointment, foam, cream, gel, paste using a syringe, bandage, transdermal patch, insert or syringe-like applicator. This is preferably in the form of a controlled release formulation or a sustained release formulation, which is administered, for example, intradermally or subcutaneously locally or directly injected into the skin adjacent to or within the area to be treated. The active composition can also be delivered by iontophoresis. Preservatives can be used to prevent the growth of fungi and other microorganisms. Suitable preservatives include, but are not limited to, benzoic acid, butylparaben, ethylparaben, methylparaben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenethyl alcohol, thimerosal and combinations thereof.

[0379] A pharmaceutical composition containing a medicament can be formulated as an emulsion for topical application. An emulsion contains a liquid distributed in a second liquid. The emulsion can be an oil-in-water emulsion or a water-in-oil emulsion. Either or both of the oil phase and the water phase can contain one or more surfactants, emulsifying agents, emulsion stabilizers, buffering agents, and other excipients. The oil phase can contain other oily pharmaceutically approved excipients. Suitable surfactants include, but are not limited to, anionic surfactants, nonionic surfactants, cationic surfactants, and amphoteric surfactants. The composition for topical application can also include at least one suitable suspending agent, antioxidant, chelating agent, emollient, or humectant.

[0380] Ointments and creams can be formulated, for example, with an aqueous or oily base, to which suitable thickening agents and / or gelling agents are added. Lotions can be formulated with an aqueous or oily base and generally also contain one or more emulsifying agents, stabilizers, dispersing agents, suspending agents, thickening agents, or coloring agents. Liquid sprays can be delivered from a pressurized package, for example, through a specially shaped cap. Oil-in-water emulsions can also be used in compositions, patches, bandages, and articles. These systems are semi-solid emulsion, microemulsion, or foam emulsion systems.

[0381] In some embodiments, the medicaments described herein can be formulated as inhalants. The inhalation method can deliver the drug directly to the airways. The medicaments can be formulated as aerosols, microspheres, liposomes, or nanoparticles. The medicaments can be formulated with solvents, gases, nitrates, or any combination thereof. The compositions described herein are optionally formulated as liquid aerosols or inhalable dry powders for delivery. Liquid aerosol formulations are optionally atomized primarily into particle sizes that can be delivered to the terminal bronchioles and respiratory bronchioles. Liquid aerosol and inhalable dry powder formulations are preferably delivered through the entire intrapulmonary tree to the terminal bronchioles and ultimately to the parenchymal tissue.

[0382] The aerosolized formulations described herein are optionally delivered using an aerosol-forming device (such as a nebulizer, vibrating perforated plate, or ultrasonic nebulizer), preferably selected to allow the formation of aerosol particles with a mass median average diameter mainly between 1 and 5 μm. In addition, the formulations preferably have a balanced osmotic pressure, ionic strength, and chloride concentration, and a minimum aerosolizable volume capable of delivering an effective dose of the medicament. In addition, the aerosolized formulations preferably have no negative impact on airway function and do not cause undesirable side effects.

[0383] A nebulizing device suitable for administering the aerosol formulations described herein includes, for example, jet, vibrating porous plate, ultrasonic nebulizers, and electrically powered dry powder inhalers, which are capable of nebulizing the formulation into an aerosol particle size predominantly in the range of 1-5 μm. By "predominantly" in this application is meant that at least 70% but preferably more than 90% of all the aerosol particles produced are in the range of 1-5 μm. The jet nebulizer operates by breaking up a liquid solution into aerosol droplets by means of air pressure. The vibrating porous plate nebulizer operates by using acoustic vacuum generated by a rapidly vibrating porous plate to extrude solvent droplets through the porous plate. The ultrasonic nebulizer operates by using a piezoelectric crystal to shear the liquid into small aerosol droplets. A variety of suitable devices are available, including, for example, the AeroNeb™ and AeroDose™ vibrating porous plate nebulizers (AeroGen, Inc., Sunnyvale, California), nebulizers (Medic-Aid Ltd., West Sussex, England), Pari Pari LC jet nebulizers (Pari Respiratory Equipment, Inc., Richmond, Virginia) and the Aerosonic™ (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany) and (Omron Healthcare, Inc., Vernon Hills, Illinois) ultrasonic nebulizers.

[0384] In some embodiments, the medicament can be formulated with an oily matrix or ointment to form a semisolid composition having a desired shape. In addition to the medicament, these semisolid compositions can contain dissolved and / or suspended bactericides, preservatives, and / or buffer systems. The petrolatum component that can be included can be any petrolatum having a viscosity range from mineral oil admixed with isobutylene, colloidal silica, or stearate to paraffin wax. Absorbent matrices can be used with the oily systems. Additives can include cholesterol, lanolin (lanolin derivatives, beeswax, fatty alcohols, lanolin alcohols, low HLB (hydrophilic-lipophilic balance) emulsifiers, and a variety of ionic and nonionic surfactants, either alone or in combination.

[0385] Controlled-release or sustained-release transdermal or topical formulations can be achieved by adding gradually releasing additives available in the art, such as polymeric structures, matrices. For example, the composition can be administered by using hot melt extrusion products, such as bioadhesive hot melt extrusion films. The formulation can comprise a crosslinked polycarboxylic acid polymer formulation. The crosslinking agent can be present in an amount sufficient to provide sufficient adhesion to allow the system to remain attached to the target epithelial or endothelial cell surface for a sufficient time to allow the desired release of the compound.

[0386] Inserts, transdermal patches, bandages or articles can comprise a polymer blend or coating that provides for the release of the medicament at a constant rate over a longer period of time. In some embodiments, the article, transdermal patch or insert comprises a water-soluble pore former, such as polyethylene glycol (PEG), which can be mixed with a water-insoluble polymer to increase the durability of the insert and extend the release of the active ingredient.

[0387] Transdermal devices (inserts, patches, bandages) can also comprise water-insoluble polymers. Rate-controlling polymers can be useful for administration to sites where pH changes can be used to affect release. These rate-controlling polymers can be applied during the spray and drying process with the active compound using a continuous coating film. In one embodiment, a coating formulation is used to coat pellets comprising the active ingredient, and these pellets are compressed to form solid, biodegradable inserts.

[0388] Polymeric formulations can also be used to provide controlled or sustained release. Bioadhesive polymers described in the art can be used. By way of example, sustained-release gels and compounds can be incorporated into a polymer matrix, such as a hydrophobic polymer matrix. Examples of polymer matrices include microparticles. The microparticles can be microspheres, and the core can be of a different material than the polymer shell. Alternatively, the polymer can be cast into slabs or films, powders or gels (such as hydrogels) produced by milling or other standard techniques. The polymer can also be in the form of a coating or part of a bandage, stent, catheter, vascular graft or other device to facilitate the delivery of the medicament. The matrix can be formed by solvent evaporation, spray drying, solvent extraction and other methods known to those skilled in the art.

[0389] Kits having a unit dose of one or more of the medicaments described herein are generally provided in oral or injectable doses. Such kits can include a container containing the unit dose, an informational package insert describing the use of the drug in treating the disease and the attendant benefits, and optionally an appliance or device for delivering the composition.

[0390] Treatment methods

[0391] In one aspect, the present disclosure provides a method of treating a subject having a disease or disorder. In some cases, treating a subject having a disease or disorder includes administering to a subject in need thereof a compound or salt of any one of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III), formula (III-A), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), formula (IV-D), formula (V), formula (VI), formula (VII), formula (VIII), or a pharmaceutical composition thereof. In some cases, the disease or disorder is cancer. In some cases, the disease or disorder is a tumor.

[0392] In one aspect, the present disclosure provides a method of treating a subject having a tumor. In some cases, treating a subject having a tumor includes administering to a subject in need thereof a compound or salt of any one of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III), formula (III-A), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), formula (IV-D), formula (V), formula (VI), formula (VII), formula (VIII), or a pharmaceutical composition thereof. In some cases, the tumor is associated with cancer. In some cases, the cancer is selected from lung cancer, kidney cancer, urethral cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, gastric cancer, and esophageal cancer.

[0393] In one aspect, the present disclosure provides a method of treating a subject having cancer. In some cases, treating a subject having cancer includes administering to a subject in need thereof a compound or salt of any one of formula (I*), formula (I), formula (I-A), formula (I-B), formula (II), formula (II-A), formula (III), formula (III-A), formula (IV), formula (IV-A), formula (IV-B), formula (IV-C), formula (IV-D), formula (V), formula (VI), formula (VII), formula (VIII), or a pharmaceutical composition thereof. In some cases, the cancer is selected from lung cancer, kidney cancer, urethral cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, gastric cancer, and esophageal cancer.

[0394] In some embodiments, the compounds described herein can be used to treat diseases, such as, but not limited to, hyperproliferative diseases, including: cancers of the head and neck, including tumors of the head, neck, nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx, hypopharynx, salivary glands, and paraganglioma; cancers of the liver and biliary system, particularly hepatocellular carcinoma; bowel cancer, particularly colorectal cancer; ovarian cancer; small cell and non-small cell lung cancers (SCLC and NSCLC); breast sarcomas, such as fibrosarcoma, malignant fibrous histiocytoma, embryonal rhabdomyosarcoma, leiomyosarcoma, neurofibrosarcoma, osteosarcoma, synovial sarcoma, liposarcoma, and alveolar soft part sarcoma; leukemias, such as acute promyelocytic leukemia (APL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), and chronic myeloid leukemia (CML); neoplasms of the central nervous system, particularly brain cancer; multiple myeloma (MM); lymphomas, such as Hodgkin lymphoma, lymphoplasmacytic lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue lymphoma, mantle cell lymphoma, B-cell large cell lymphoma, Burkitt lymphoma, and T-cell anaplastic large cell lymphoma. Clinically, practicing the methods described herein and using the compositions described herein will result in a decrease in the size or number of cancerous growths and / or alleviation of related symptoms (where applicable). Pathologically, practicing the methods described herein and using the compositions described herein will produce a pathologically relevant response, such as: inhibition of cancer cell proliferation, reduction in the size of the cancer or tumor, prevention of further metastasis, and inhibition of tumor angiogenesis. Methods of treating such diseases include administering to a subject a therapeutically effective amount of the combination of the present invention. The method can be repeated as needed. The cancer can be renal cancer, lung cancer, gastric cancer, or ovarian cancer.

[0395] In some embodiments, treatment of a tumor-bearing subject inhibits tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and still even more preferably at least about 80% relative to an untreated subject. A therapeutically effective amount of the therapeutic compound can reduce the size of the tumor or otherwise improve the symptoms of the subject, which is typically a human, but can be another mammal.

[0396] In some embodiments, the compounds described herein can be administered in combination with other therapeutic agents, including antibodies, alkylating agents, angiogenesis inhibitors, antimetabolites, DNA cleavage agents, DNA cross-linking agents, DNA intercalating agents, DNA minor groove binders, enediynes, heat shock protein 90 inhibitors, histone deacetylase inhibitors, immunomodulators, microtubule stabilizers, nucleoside (purine or pyrimidine) analogs, nuclear export inhibitors, proteasome inhibitors, topoisomerase (I or II) inhibitors, tyrosine kinase inhibitors, and serine / threonine kinase inhibitors. Specific therapeutic agents include adalimumab, ansamitocin P3, auristatins, bendamustine, bevacizumab, bicalutamide, bleomycin, bortezomib, busulfan, callistatin A, camptothecin, capecitabine, carboplatin, carmustine, cetuximab, cisplatin, cladribine, cytarabine, cryptophycins, dacarbazine, dasatinib, daunorubicin, docetaxel, doxorubicin, duocarmycin, dynemycin A, epothilones, etoposide, floxuridine, fludarabine, 5-fluorouracil, gefitinib, gemcitabine, ipilimumab, hydroxyurea, imatinib, infliximab, interferons, interleukins, β-lapachone, lenalidomide, irinotecan, maytansine, mechlorethamine, melphalan, 6-mercaptopurine, methotrexate, mitomycin C, nilotinib, oxaliplatin, paclitaxel, procarbazine, suberoylanilide hydroxamic acid (SAHA), 6-thioguanine, thiotepa, teniposide, topotecan, trastuzumab, trichostatin A, vinblastine, vincristine, and vindesine.

[0397] The compounds described herein can be used to prepare a medicament for preventing or treating a disease or condition. Further, a method for treating any disease or condition described herein in a subject in need of such treatment involves administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one of the compounds described herein or a pharmaceutically acceptable salt, pharmaceutically acceptable prodrug, or pharmaceutically acceptable solvate thereof.

[0398] Compositions containing the compounds described herein can be administered for prophylactic and / or therapeutic treatment. In therapeutic applications, the composition is administered to a patient who has a disease or condition in an amount sufficient to cure or at least partially alleviate the symptoms of the disease or condition. The amount effective for this use will depend on the severity and course of the disease or condition, previous treatment, the health status, weight, response to the drug, and the judgment of the attending physician of the patient.

[0399] In prophylactic applications, a composition containing a compound described herein is administered to a patient who is predisposed to, or at risk of developing, a particular disease, disorder, or condition. Such an amount is defined as a "prophylactically effective amount or dose". In this use, the exact amount also depends on the patient's health, weight, etc. When used in a patient, the amount effective for this use will depend on the severity and course of the disease, disorder, or condition, previous treatment, the patient's health and response to the drug, and the judgment of the attending physician.

[0400] In the event that the patient's condition does not improve, the compound may be administered chronically, i.e., over a prolonged period of time, including throughout the patient's lifetime, at the discretion of the physician administering the compound, in order to ameliorate or otherwise control or limit the symptoms of the patient's disease or condition.

[0401] Once improvement of the patient's condition has occurred, a maintenance dose is administered as necessary. Subsequently, the dose or the frequency of administration, or both, may be reduced according to the symptoms, until the disease, disorder, or condition has improved and is maintained at a certain level. However, the patient may require intermittent treatment on a long-term basis whenever any symptoms recur.

[0402] The amount of a given pharmaceutical agent corresponding to this amount will vary depending on a variety of factors, such as the particular compound, the disease or condition and its severity, the identity of the subject or host to be treated (e.g., weight), but can still be determined in a manner recognized in the art, based on the particular circumstances surrounding the case, including, for example, the particular pharmaceutical agent administered, the route of administration, the condition being treated, and the subject or host being treated. However, generally speaking, the dose for adult treatment is usually in the range of about 0.02 to about 5000 mg per day, and in some embodiments, about 1 to about 1500 mg per day. The desired dose may be conveniently administered in a single dose or divided doses simultaneously (or over a short period of time) or at appropriate intervals, such as two, three, four, or more divided doses per day.

[0403] The pharmaceutical compositions described herein may be in unit dosage forms suitable for precise single-dose administration. In a unit dosage form, the preparation is divided into unit doses containing a suitable amount of one or more compounds. The unit dose may be in a package form containing a discrete amount of the preparation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. An aqueous suspension composition may be packaged in a single-dose non-resealable container. Alternatively, a multi-dose resealable container may be used, in which case a preservative is usually included in the composition. By way of example only, a preparation for parenteral injection may be presented in unit dosage form, which includes, but is not limited to, an ampoule or in a multi-dose container, with a preservative added.

[0404] The toxicity and therapeutic efficacy of such therapeutic regimens can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, including but not limited to determining the LD 50 (the dose lethal to 50% of the population) and the ED 50 (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index, and it can be expressed as the ratio of LD 50 to ED 50 . Compounds preferably exhibiting a high therapeutic index are selected. Data obtained from cell culture assays and animal studies can be used to formulate a dosage range for human use. The dosage of such compounds is preferably within the circulating concentration range that includes the ED 50 and is minimally toxic. The dosage can vary within this range depending on the dosage form employed and the route of administration utilized.

[0405] In certain embodiments, the present invention provides a method of treating or preventing a disease, condition, or disorder in a patient in need thereof, comprising administering to the patient an effective amount of a compound of any embodiment of the present invention or a pharmaceutically acceptable salt thereof. The disease, condition, or disorder can be selected from the groups described elsewhere herein.

[0406] Preparation of Compounds

[0407] The compounds of the present disclosure can generally be prepared by a variety of methods well known to those skilled in the art of organic synthesis. By way of example, the compounds of the present disclosure can be synthesized using the methods described herein as well as synthetic methods known in the art of synthetic organic chemistry or variants thereof as understood by those skilled in the art. The compounds of the present disclosure can be prepared according to the schemes and examples described elsewhere herein.

[0408] The following examples further illustrate the invention, but should of course not be construed as limiting its scope in any way.

[0409] Examples

[0410] The following synthetic schemes are provided for illustrative purposes only and not for limitation. The following examples illustrate various methods for preparing the compounds described herein. It should be understood that those skilled in the art will be able to prepare these compounds by similar methods or by combining other methods known to those skilled in the art. It should also be understood that those skilled in the art will be able to prepare in a manner similar to that described below by using appropriate starting materials and modifying the synthetic routes as needed. Generally, the starting materials and reagents can be obtained from commercial suppliers or synthesized from sources known to those skilled in the art or prepared as described herein. The following abbreviations, as used below and throughout the specification of the present invention, should be understood to have the following meanings unless otherwise indicated:

[0411]

[0412]

[0413] TCEP Tris(2 - carboxyethyl)phosphine

[0414] Example 1: Synthesis of Compound 24

[0415]

[0416] Compound 24: To a solution of Compound 21 (mesylate, 53 mg, 0.1 mmol) in DMF (2 mL) was added formaldehyde (30% aqueous solution, 0.1 mL), and then NaBH(OAc)3 (64 mg) was added. The reaction was stirred at room temperature for 2 h. Then, the mixture was directly purified by reverse - phase high - performance liquid chromatography (RP - HPLC) to give Compound 24 (TFA salt, 52 mg) as a yellow solid. MS: 464.3 [M + H] + 。

[0417] Example 2: Synthesis of Compound 23

[0418]

[0419] Compound 22: To a suspension of Compound 19 (121 mg, 212 μmol) in anhydrous DCM (12 mL) was added a solution of thionyl chloride in DCM (1 M, 635 μL, 635 μmol). The mixture was stirred at room temperature for 10 min, and then the solvent was removed by decantation, leaving a gummy solid of crude chloride 20. The solid was washed with ether, dried under vacuum, and redissolved in anhydrous DMF (1 mL). To the resulting solution were added irinotecan mesylate (21) (113 mg, 212 μmol), tetrabutylammonium iodide (4 mg, 106 μmol), and DIEA (130 μL, 748 μmol). The resulting mixture was heated at 60 °C for 16 h. Then the mixture was purified by reverse - phase high - performance liquid chromatography (RP - HPLC). The fractions were lyophilized to give Compound 22 (18 mg) as a white solid.

[0420] Compound 23: To a solution of Compound 22 (TFA salt, 16.6 mg, 15 μmol) in DMF (1 mL) were added DIEA (10 μL, 58 μmol) and methyl iodide (1 mL). The mixture was stirred at 50 °C for 24 h, and then purified by reverse - phase high - performance liquid chromatography (RP - HPLC). The fractions were lyophilized to give Compound 23 (1.7 mg) as a white solid. MS: 1018.7 [M + ].

[0421] Example 3: Preparation of Antibody - Drug Conjugate (ADC) - 1.

[0422]

[0423] , where the DAR is 4.3.

[0424] Adjust the pH or buffer exchange of 55 mg of the UC-961 antibody in the formulation buffer to a formulation of 5% (v / v) 0.5 M Tris, 0.025 M EDTA, pH 8.5 for reduction. 0.025 M EDTA was added to the formulation buffer to prevent metal-catalyzed disulfide reoxidation. 10 mM TCEP (7 equivalents per antibody) was added and incubated at 20 °C for 2 h to reduce the targeted number of interchain disulfide bonds and generate the average number of free thiols required per antibody. The free thiols were conjugated by adding a DMA solution of 20 mM drug linker 23 (12 equivalents) and stirring at 20 °C for 1 h. Additional DMA solvent was added before adding the drug linker to maintain the solubility of the drug linker after addition and mixing was carried out using a stirred flask. The conjugation was terminated by adding an excess of NAC (110 mM) and stirring was continued for 30 min to quench the unreacted maleimide. Incubate with activated carbon at 15 rpm for 1 h at room temperature and / or filter through to remove the excess quenched drug linker. Exchange ADC-1 (yield 91 - 94%) into the final formulation buffer (0.1 M Arg / PBS), filter through a 0.2 μm PES, and then aliquot and store at ≤ -60 °C.

[0425] Characterization of the Conjugate

[0426] Example 4: Plasma Stability

[0427] The stability of ADC-1 in mouse, cynomolgus monkey, and human plasma was tested.

[0428] Incubate ADC-1 at a concentration of 50 μg / mL and a volume of 300 μL in IgG-depleted plasma. IgG depletion was performed using a HiTRap Protein G column (Cytiva). Incubate the samples in Eppendorf tubes for time periods of 0, 1, 3, 7, and 15 days at 37 °C. After incubation at the appropriate time, transfer the samples to an -80 °C freezer until they can be processed. The samples were incubated twice at each time point.

[0429] To isolate ADC-1 from plasma, 100 μL of sample was mixed with 100 μL of Protein A magnetic bead slurry (Thermo Pierce) and 900 μL of sodium phosphate (50 mM, pH 7) and shaken at room temperature for 2 hours. The beads were then washed with 1 mL of 0.1% Triton, 0.1% IPA to remove non-specific binding and then washed twice in 1 mL of PBS for 30 minutes each time with shaking. Then, ADC-1 was eluted from the beads in 100 μL of low pH, high organic mix (40 mM glycine, 2% formic acid, 50% acetonitrile) for 60 minutes. The bead slurry was then centrifuged and 50 μL of the supernatant was injected into LC-MS.

[0430] LC was performed using a 50 mm x 2.1 bioZen XB-C8 column with a gradient of H2O and acetonitrile (each with 0.1% formic acid) over seven minutes. MS was operated in the intact protein mode where the source temperature was 500 °C, the extinction voltage was 200 V, and the mass range was 900 - 4500 m / z.

[0431] Processing was performed in Sciex OS and the stability results are as Figure 1 shown.

[0432] Example 5: Cell Binding

[0433] The ability of ADC-1 to bind to Jeko-1 cells was measured using an in vitro cell binding assay.

[0434] For each Jeko-1 cell line, 500,000 cells were plated at 50 μL per well in a 96-well deep well plate (Thermo Scientific #249946). Starting from a 1-μg / mL stock solution, a 1:3 dilution series of the primary antibody was prepared. Subsequently, 50 μL of the primary antibody at different concentrations (final concentrations ranging from 17 pg / mL to 1000 ng / mL) was placed in the 50 μL containing the cells. The cells and the antibody were mixed and incubated on ice for 20 minutes. For the first wash: 300 μL of FACS buffer was added and centrifuged at 500 × g for 5 minutes at 4 °C. The supernatant was discarded and the cells were resuspended in 400 μL of FACS buffer for additional washes. After the second wash, the cells were resuspended in 100 μL of goat anti-human IgG secondary antibody phycoerythrin (PE) (ThermoFisher Scientific #12-4998-82) at a final concentration of 1 μg / mL and incubated on ice (in the dark) for 20 minutes. The cells were washed twice as previously described and analyzed on a BD FACSVerse using Flowjo software (version 10). The percentage of maximum binding relative to the highest concentration was plotted and the half-maximal effective concentration (EC 50 ) was determined using GraphPad Prism version 7.0, and the binding percentage of the ADC was plotted, in Figure 2 .

[0435] Example 6: Cytotoxicity measurement

[0436] The ability of ADC-1 to inhibit cell growth was measured using an in vitro cytotoxicity assay.

[0437] Cells were cultured in the logarithmic growth phase and aliquoted into 96-well plates. Each cell line was plated at slightly different concentrations, but in the range of 5x10 3 to 50x10 4 cells / well. The cells were incubated in duplicate with 3-fold serial dilutions (3000, 1000, 333, 111, 37, 12.3, 4.1, 1.37, 0.46, 0.15 nM) of a specific immunoconjugate starting at 3000 or 1000 nanomoles at 37 °C and 5% CO2 for 72 hours. After treatment, the cells were incubated with an equal volume of CellTiter- reagent (Promega Inc.) for 15 minutes at room temperature and the viability was determined using a photometer. Table 1 shows the EC50 values of ADC-1 in different cell lines.

[0438] Table 1.

[0439]

[0440] Example 7: Internalization

[0441] Harvest Jeko-1 cells, wash with cold PBS, and resuspend in cold FACS buffer (containing PBS and 2% FBS) at a concentration of 1×10 7 cells / mL. Aliquots of 1×10 6 cells are added to microcentrifuge tubes or wells. Dilute the primary antibody to produce 10-fold stock solutions of 300 μg / mL or 1 mg / mL to allow 10 μl of each solution to be added to the appropriate tubes.

[0442] The control group consists only of unstained secondary antibody (goat anti-human IgG-PE, Fc-γ specific)

[0443] (Thermo Fisher Scientific #12-4998-82). The test groups contain cells treated under the following conditions and are evaluated with primary antibody at 30 μg / mL (203 nM) or 100 μg / mL (676 nM) according to the experiment: Except for the control, the cells are placed on ice for 20 minutes, the control group is centrifuged at 300×g for 4 minutes, washed twice with 200 μl of FACS buffer, resuspended in 100 μl of FACS buffer, and incubated at 37 °C for 30, 60, 120, or 240 minutes. After incubation, the cells are centrifuged at 250×g, washed twice with FACS buffer, and resuspended in 100 μl of FACS buffer. Dilute the 10-fold secondary antibody stock solution 1:2000 in FACS buffer and add 10 μl per tube to the appropriate tubes. Incubate the cells on ice for 20 minutes, wash twice with FACS buffer, and resuspend in 100 μl of fixation buffer (4% paraformaldehyde in PBS). Then perform FACS analysis to evaluate the median fluorescence intensity (MFI). The internalization results are as Figure 3 shown.

[0444] Example 8: DAR by RP-HPLC

[0445] Obtain the drug-to-antibody ratio (DAR) of an ADC (e.g., ADC-1) using reverse-phase HPLC.

[0446] RP-HPLC conditions: Column – Phenomenex Kinetex 50x4.6 mm, 2.6 μm, part number: PL1912-1502. MPA – 0.1% TFA / H20, MPB – 0.1% TFA / ACN

[0447] Method: Flow rate – 1 mL / min, gradient – see Table 2. Column temperature – 50 °C.

[0448] Sample temperature – RT. DAD 214 nm, BW 16 nm; reference 440 nm, BW 80 nm; peak width > 0.4 min (8 s response time (0.62 Hz); spectrum: 200–600 nm, step 1.2 nm, slit 8 nm.

[0449] Sample – pure injection, ~5 μg.

[0450] Table 2: RP-HPLC method flow rate gradient

[0451] Time (minutes) B％ 0 30 2 30 10 41 11.5 90 15.5 90 16.5 30 20 30

[0452] Obtain ADC-1 with a DAR of 4.3 and a monomer purity of 93%.

Claims

1. A compound of formula (I*): or a pharmaceutically acceptable salt thereof, wherein; L and M are linkers; n and u are selected from 0 and 1; R 1A selected from hydrogen and C 1-6 alkyl; R 1B is an optionally substituted C 1-6 alkyl group, wherein said C 1-6 alkyl group is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NH2 and oxo; wherein when u is 1, R 1B is absent or is -C 1-6 alkylene-O-; and wherein either u or n is 1.

2. The compound or salt according to claim 1, wherein formula (I*) is represented as or a pharmaceutically acceptable salt thereof, wherein; R 1A selected from C 1-6 alkyl; R 1B selected from C 1-6 alkyl; and L is a linker.

3. The compound or salt according to claim 1 or 2, wherein said L is L 1 -L 2 -L 3 -L 4 -(L 5 ) m -R 2 ; and L 1 selected from C 1-6 alkylene; L 2 selected from optionally substituted C 5-6 carbocyclic ring, wherein said C 5-6 carbocyclic ring is optionally substituted by one or more substituents independently selected from R 3 ; L 3 Residues selected from residues of 1 to 7 amino acids; L 4 selected from optionally substituted C 1-6 alkylene, wherein said C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl; L 5 selected from (-O-CH2-CH2-) q ; R 2 selected from optionally substituted 5- to 6-membered heterocycles, wherein the 5- to 6-membered heterocycle is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl; Each R 3 is independently selected from sugar; m is selected from 0 and 1; and q is selected from 1 to 10.

4. The compound or salt according to any one of claims 1 to 3, wherein R 1A is methyl and R 1B is methyl.

5. The compound or salt according to any one of claims 1 to 4, wherein formula (I*) and formula (I) are each represented by formula (I-B): or a pharmaceutically acceptable salt thereof, wherein; b is selected from 0 to 1.

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

7. The compound or salt according to claim 6, wherein R 3 is 8. The compound or salt according to claim 7, wherein R 3 is 9. The compound or salt according to claim 5, wherein b is 0.

10. The compound or salt according to any one of claims 1 to 9, wherein L 3 is selected from residues comprising from 1 to 5 amino acids.

11. The compound or salt according to any one of claims 1 to 10, wherein the amino acid is a natural amino acid.

12. The compound or salt according to any one of claims 1 to 10, wherein the amino acid is selected from α-amino acids and β-amino acids.

13. The compound or salt according to claim 12, wherein the amino acid is selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, citrulline, and β-alanine.

14. The compound or salt according to claim 13, wherein the amino acid is selected from β-alanine, citrulline, and valine.

15. The compound or salt according to any one of claims 1 to 14, wherein L 3 is selected from 16. The compound or salt according to any one of claims 1 to 10, wherein the amino acid is a non-natural amino acid.

17. The compound or salt according to any one of claims 1 to 16, wherein L 4 is selected from C 1-6 alkylene substituted by one oxo.

18. The compound or salt according to claim 17, wherein L 4 is selected from 19. The compound or salt according to any one of claims 1 to 18, wherein m is 0.

20. The compound or salt according to any one of claims 1 to 18, wherein m is 1.

21. The compound or salt according to claim 20, wherein q is 2.

22. The compound or salt according to any one of claims 1 to 21, wherein R 2 is selected from optionally substituted 5-membered heterocycles.

23. The compound or salt according to claim 22, wherein R 2 is 24. The compound or salt according to any one of claims 1 to 23, wherein n is 1.

25. The compound or salt according to any one of claims 1 to 24, wherein the compound is selected from or a pharmaceutically acceptable salt of any of them.

26. The compound or salt according to claim 25, wherein the compound is selected from or a pharmaceutically acceptable salt of any of them.

27. The compound or salt according to any one of claims 1 to 26, further comprising a ligand.

28. The compound or salt according to any one of claims 1 to 27, wherein the compound or salt is further modified by a ligand.

29. The compound or salt according to any one of claims 1 to 28, wherein the compound or salt is covalently attached to a ligand.

30. The compound or salt according to any one of claims 1 to 28, wherein the compound or salt reacts to form a covalent bond with a ligand.

31. The compound or salt according to any one of claims 1 to 30, wherein formula (I*) or formula (I) is represented by formula (II): or a pharmaceutically acceptable salt thereof, wherein; Lg is the ligand.

32. The compound or salt according to claim 31, wherein formula (II) is or a pharmaceutically acceptable salt thereof.

33. The compound or salt according to any one of claims 27 to 32, wherein the ligand is selected from antibodies and antigen-binding fragments thereof.

34. The compound or salt according to claim 33, wherein the ligand is selected from chimeric antibodies, humanized antibodies, and human antibodies.

35. A compound of formula (VII): or a pharmaceutically acceptable salt thereof.

36. The compound or salt according to claim 35, wherein the compound is or a pharmaceutically acceptable salt thereof.

37. A compound of formula (III): or a pharmaceutically acceptable salt thereof, wherein; R 1A selected from C 1-6 alkyl; R 1B is an optionally substituted C 1-6 alkyl group, wherein said C 1-6 alkyl group is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NH2 and oxo; L is a linker; and Lg is a ligand.

38. The compound or salt according to claim 37, wherein the linker is L 1 -L 2 -L 3 -L 4 -(L 5 ) m -R 2 ; and L 1 Selected from C 1-6 alkylene; L 2 selected from optionally substituted C 5-6 carbocyclic ring, wherein said C 5-6 carbocyclic ring is optionally substituted by one or more substituents independently selected from R 3 ; L 3 a residue selected from residues of from 1 to 7 amino acids; L 4 selected from optionally substituted C 1-6 alkylene, wherein said C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl; L 5 selected from (-O-CH2-CH2-) q ; R 2 selected from optionally substituted 5- to 6-membered heterocycles, wherein the 5- to 6-membered heterocycle is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl; Each R 3 is independently selected from sugar; m is selected from 0 and 1; and q is selected from 1 to 10.

39. The compound or salt according to claim 38, wherein formula (III) is or a pharmaceutically acceptable salt thereof.

40. The compound or salt according to any one of claims 37 to 39, wherein the ligand is selected from an antibody or an antigen-binding fragment thereof.

41. The compound or salt according to claim 40, wherein the ligand is selected from a chimeric antibody, a humanized antibody, and a fully human antibody.

42. The compound or salt according to claim 1, wherein formula (I*) is represented as or a pharmaceutically acceptable salt thereof, wherein; R 1A selected from hydrogen and C 1-6 alkyl; R 1B is absent or -C 1-6 alkylene-O-; M is (L 2 ) x -(L 2B ) z -(L 2C ) y -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 ; L 2 Selected from C 1-6 alkylene; L 2B Selected from (NR 4 ) t C(O)O-CH2-phenyl, wherein said phenyl is optionally substituted with one or more R 5 substituents; L 2C Selected from C(O)O-CH2-phenyl, wherein said phenyl is optionally substituted with one or more R 6 substituents; L 3 a residue selected from residues of from 1 to 7 amino acids; L 4 Selected from optionally substituted C 1-6 alkylene, wherein said C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl; L 5 Selected from (O-CH2-CH2) q -(NR 3 ) s ; L 6 selected from optionally substituted C 1-6 alkylene, wherein said C 1-6 alkylene is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl; L 7 Selected from C 5-6 carbocyclic ring; R 2 selected from optionally substituted 5- to 6-membered heterocycles, wherein the 5- to 6-membered heterocycle is optionally substituted by one or more substituents independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl; R 3 selected from hydrogen and C 1-6 alkyl; R 4 selected from hydrogen and optionally C 1-6 alkyl substituted by one or more SO2C 1-6 alkyl; Each R 5 is independently selected from sugars; Each R 6 is independently selected from halogen, -OH, -CN, -NO2, -NH2, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl; m is selected from 0 and 1; p is selected from 0 and 1; q is selected from 0 to 8; s is selected from 0 and 1; t is selected from 0 and 1; x is selected from 0 and 1; y is selected from 0 and 1; and z is selected from 0 and 1.

43. The compound or salt according to claim 1 or 42, wherein formula (I*) or formula (IV) is represented by formula (IV-A): or a pharmaceutically acceptable salt thereof; wherein R 1A is selected from hydrogen and methyl.

44. The compound or salt according to claim 1 or 42, wherein formula (I*) or formula (IV) is represented by formula (IV-B): or a pharmaceutically acceptable salt thereof.

45. The compound or salt according to any one of claims 1, 42, 43 or 44, wherein M is L 2 -(L 2B ) z -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 。 46. The compound or salt according to claim 45, wherein M is L 2 -L 3 -L 4 -R 2 。 47. The compound or salt according to claim 45, wherein M is L 2 -L 3 -L 4 -L 7 -R 2 .

48. The compound or salt according to claim 45, wherein M is L 2 -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 。 49. The compound or salt according to claim 45, wherein M is L 2 -L 2B -L 3 -L 4 -R 2 .

50. A compound or salt according to any one of claims 43 to 49, wherein L 2 is a C1 alkylene group.

51. The compound or salt according to claim 1, 42, 43 or 44, wherein M is L 2B -L 3 -L 4 -R 2 and t is 0.

52. A compound or salt according to claim 1, 42, 43 or 44, wherein M is L 2C -L 3 -L 4 -R 2 .

53. A compound or salt according to claim 1, 42, 43 or 44, wherein M is L 2C -L 3 -L 4 -L 7 -R 2 。 The compound or salt according to claim 1, 42, 43 or 44, wherein M is L 2C -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 。 55. A compound or salt according to any one of claims 1, 42, 43, 44 or 52 to 54, wherein L 2C is C(O)O-CH2-phenyl.

56. The compound or salt according to any one of claims 1 to 55, wherein L 3 is selected from residues comprising 1 to 5 amino acids.

57. The compound or salt according to any one of claims 1 to 56, wherein the amino acid is a natural amino acid.

58. The compound or salt according to any one of claims 1 to 57, wherein the amino acid is selected from α-amino acids and β-amino acids.

59. The compound or salt according to claim 58, wherein the amino acid is selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, citrulline, sarcosine, and β-alanine.

60. The compound or salt according to any one of claims 1 to 59, wherein L 3 said amino acid of is selected from glycine and phenylalanine.

61. The compound or salt according to any one of claims 1 to 60, wherein L 3 the residue of which comprises 4 amino acids.

62. A compound or salt according to any one of claims 1 to 50 or 52 - 61, wherein the residue of L 3 comprises at least two different amino acids.

63. The compound or salt according to any one of claims 1 to 50 or 52 - 61, wherein L 3 is 64. The compound or salt according to claim 51, wherein L 3 said amino acid of is selected from sarcosine.

65. The compound or salt according to claim 64, wherein L 3 the residue of contains 1 amino acid.

66. The compound or salt according to claim 65, wherein L 3 is 67. The compound or salt according to claim 1 or 51, wherein L 2B is selected from (NR 4 ) t C(O)O-C1-alkylene-phenyl, wherein said phenyl is substituted by one R 5 ; t is 1.

68. The compound or salt according to claim 67, wherein R 5 is 69. The compound or salt according to any one of claims 1 to 68, wherein L 4 is selected from -C(O)-(CH2)2- and -C(O)-(CH2)5-.

70. The compound or salt according to claim 1 or 48, wherein L 5 is -(O-CH2-CH2-)4-NH-.

71. The compound or salt according to claim 1 or 48, wherein L 6 is selected from -C(O)-(CH2)2- and -C(O)-(CH2)5-.

72. The compound or salt according to claim 71, wherein L 6 is selected from -C(O)-(CH2)2-.

73. A compound or salt according to any one of claims 1, 47 or 48, wherein L 7 is phenyl.

74. The compound or salt according to any one of claims 1 to 73, wherein R 2 is selected from optionally substituted 5-membered heterocycles.

75. The compound or salt according to claim 74, wherein R 2 is selected from 5-membered heterocycles substituted with at least two oxo groups.

76. The compound or salt according to claim 75, wherein R 2 is 77. The compound or salt according to claim 1, 42 or 43, wherein M is selected from 78. A compound or salt according to claim 1, 42, 43 or 44, wherein L is selected from L 2C -L 3 -L 4 -R 2 , L 2C -L 3 -L 4 -L 7 -R 2 and L 2C -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 ; where L 2C Selected from 79. The compound or salt according to claim 1, 42 or 43, wherein the compound is 80. The compound or salt according to claim 1, 42 or 43, wherein the compound is or a pharmaceutically acceptable salt thereof.

81. The compound or salt according to any one of claims 42 to 80, further comprising a ligand.

82. The compound or salt according to any one of claims 42 to 81, wherein the compound or salt is further modified with a ligand.

83. The compound or salt according to any one of claims 42 to 82, wherein the compound or salt is covalently attached to a ligand.

84. The compound or salt according to any one of claims 42 to 83, wherein the compound or salt reacts with a ligand to form a covalent bond.

85. The compound or salt according to claim 42 or 43, wherein formula (IV) or formula (IV-A) is represented by or a pharmaceutically acceptable salt thereof.

86. The compound or salt according to claim 42 or 43, wherein formula (IV) or formula (IV-A) is represented by or a pharmaceutically acceptable salt thereof.

87. The compound or salt according to any one of claims 81 to 86, wherein the ligand is selected from an antibody or an antigen-binding fragment thereof.

88. The compound or salt according to claim 87, wherein the ligand is selected from a chimeric antibody, a humanized antibody, and a fully human antibody.

89. A compound of formula (V): or a pharmaceutically acceptable salt thereof.

90. A compound of formula (VI): or a pharmaceutically acceptable salt thereof.

91. The compound or salt according to claim 1, wherein u is 0 and n is 1.

92. The compound or salt according to claim 1, 2 or 91, wherein L is L 1 -L 2 -L 3 -L 4 -R 2 ; L 1 is selected from C 1-6 alkylene; L 2 is selected from optionally substituted phenylene; L 3 is selected from residues comprising 1 to 7 amino acids; L 4 is selected from optionally substituted C 1-6 alkylene, wherein the C 1-6 alkylene is substituted with one or more substituents independently selected from oxo; R 1A is methyl, R 1B is methyl, and R 2 is maleimide.

93. A compound of formula (VII): or a pharmaceutically acceptable salt thereof.

94. A compound selected from the following: or a pharmaceutically acceptable salt thereof, wherein the DAR is selected from about 1-8.

95. The compound or salt according to claim 94, wherein the DAR is about 4.

96. A compound represented by formula (VIII): or a pharmaceutically acceptable salt thereof, wherein the DAR is selected from about 1-8, and Lg is a ligand.

97. A pharmaceutical composition comprising the compound or salt according to any one of claims 1 to 96 and a pharmaceutically acceptable excipient.

98. Use of the compound or salt according to any one of claims 1 to 96 or the pharmaceutical composition according to claim 97 in the treatment of cancer.

99. A method of treating a subject having a disease or disorder, comprising administering to the subject in need thereof a compound or salt according to any one of claims 1 to 96 or a pharmaceutical composition according to claim 97.

100. The method according to claim 99, wherein the disease or disorder is selected from lung cancer, kidney cancer, urethral cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, gastric cancer, and esophageal cancer.

101. A method of treating a subject having cancer, comprising administering to the subject in need thereof a compound or salt according to any one of claims 1 to 96 or a pharmaceutical composition according to claim 97.

102. The method according to claim 101, wherein the cancer is selected from lung cancer, kidney cancer, urethral cancer, colorectal cancer, prostate cancer, glioblastoma multiforme, ovarian cancer, pancreatic cancer, breast cancer, melanoma, liver cancer, bladder cancer, gastric cancer, and esophageal cancer.

103. A method of treating a subject having a tumor, comprising administering to the subject in need thereof a compound or salt according to any one of claims 1 to 96 or a pharmaceutical composition according to claim 97.

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