Compositions and uses thereof

Abstract

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

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 380,351, filed Oct. 20, 2022, which is hereby incorporated by reference in its entirety. BACKGROUND OF THE INVENTION

[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 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 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 ;

[0008] L 2 8] is selected from C 1-6 alkylene;

[0009] L 2B is selected from (NR 4 ) t C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted with one or more R 5 ;

[0010] L 2C is selected from C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted with one or more R 6 ;

[0011] L 3 is selected from residues comprising 1 to 7 amino acids;

[0012] 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;

[0013] L 5 Selected from (O-CH2-CH2-) q -(NR 3 ) s ;

[0014] 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;

[0015] L 7 Selected from C 5-6 carbocycle;

[0016] R 1 Selected from -O- and -NR 7 -;

[0017] R 2 Selected from optionally substituted 5- to 6-membered heterocycles, wherein said 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;

[0018] R 3 Selected from hydrogen and C 1-6 alkyl;

[0019] R 4 Selected from hydrogen and C 1-6 alkyl optionally substituted by one or more SO2C 1-6 alkyl;

[0020] Each R 5 is independently selected from sugar;

[0021] Each R 6 is 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;

[0022] R 7 is selected from hydrogen and C 1-6 alkyl;

[0023] R 8 is selected from hydrogen and hydroxy;

[0024] R 9 is selected from hydrogen and halogen, wherein at least one of R 8 or R 9 is hydrogen;

[0025] m is selected from 0 and 1;

[0026] n is selected from 0 and 1;

[0027] p is selected from 0 and 1;

[0028] q is selected from 0 to 8;

[0029] s is selected from 0 and 1;

[0030] t is selected from 0 and 1;

[0031] x is selected from 0 and 1;

[0032] y is selected from 0 and 1; and

[0033] z is selected from 0 and 1.

[0034] In some embodiments, formula (I) is represented as

[0035]

[0036] or a pharmaceutically acceptable salt thereof.

[0037] In some embodiments, formula (I) is represented as

[0038]

[0039] or a pharmaceutically acceptable salt thereof.

[0040] In some embodiments, formula (I) is represented as

[0041]

[0042] or a pharmaceutically acceptable salt thereof, wherein;

[0043] Lg is a ligand.

[0044] In some embodiments, the present disclosure provides a compound of formula (III):

[0045]

[0046] or a pharmaceutically acceptable salt thereof, wherein;

[0047] Lg is a ligand;

[0048] L 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 ;

[0049] L 2 is selected from C 1-6 alkylene;

[0050] L 2B is selected from (NR 4 ) t C(O)O-C 1-6 alkylene-phenyl, wherein the phenyl is optionally substituted with one or more R 5 substituents;

[0051] L 2C is selected from C(O)O-CH2-phenyl, wherein the phenyl is optionally substituted with halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0052] L 3 is selected from residues comprising 1 to 7 amino acids;

[0053] L 4 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 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0054] L 5 is selected from (O-CH2-CH2-) q -(NR 3 ) s ;

[0055] L 6 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 alkyl, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl;

[0056] L 7 is selected from C 5-6 carbocycle;

[0057] R 1 is selected from -O- and -NR 7 -;

[0058] R 2 is 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;

[0059] R 3 is selected from hydrogen and C 1-6 alkyl;

[0060] R 4 is selected from hydrogen and C 1-6 alkyl optionally substituted by one or more SO2C 1-6 alkyl;

[0061] Each R 5 is independently selected from sugars;

[0062] R7 Selected from hydrogen and C 1-6 alkyl;

[0063] R 8 selected from hydrogen and hydroxyl;

[0064] R 9 is selected from hydrogen and halogen, wherein R 8 or R 9 At least one of is hydrogen;

[0065] m is selected from 0 and 1;

[0066] n is selected from 0 and 1;

[0067] p is selected from 0 and 1;

[0068] q is selected from 0 to 8;

[0069] s is selected from 0 and 1;

[0070] t is selected from 0 and 1;

[0071] x is selected from 0 and 1;

[0072] y is selected from 0 and 1; and

[0073] z is selected from 0 and 1.

[0074] In some embodiments, the ligand is selected from an antibody or an antigen-binding fragment thereof.

[0075] In some cases, the ligand is selected from a chimeric antibody, a humanized antibody, and a fully human antibody.

[0076] In some embodiments, a pharmaceutical composition comprises a compound or salt of Formula (I)-(IA), (IB), (II)-(III), (III-A), (IV-A), (IV-B), (IV-C), or (IV-D), and a pharmaceutically acceptable excipient of any one thereof.

[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 (A)-(II), (IA)-(II), (IB)-(II), (III-A)-(III), (IV-A)-(IV), (IV-B)-(IV), (IV-C)-(IV), or (IV-D)-(IV), or a pharmaceutical composition of any one 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-A), formula (I-B), formula (II), formula (III), formula (III-A), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D), or a pharmaceutical composition of any of the foregoing.

[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. In the event of a conflict between the incorporated publications, patents, or patent applications and the disclosure contained herein, the specification is intended to supersede and / or take precedence over any such conflicting material. Detailed Description

[0081] The following description sets forth numerous exemplary configurations, methods, parameters, etc. However, it should be recognized that such description is not intended to limit the scope of the present disclosure, but rather is provided as a description of exemplary embodiments.

[0082] In the following description, certain specific details are set forth in order to provide a thorough understanding of the various embodiments of the present disclosure. However, one of ordinary skill in the art will understand that the present disclosure may be practiced without these details.

[0083] Definitions

[0084] 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.

[0085] "Alkyl" refers to a straight-chain or branched-chain hydrocarbon chain group consisting only of carbon and hydrogen atoms, having 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 from one to eight carbon atoms (i.e., C1-C8 alkyl). In other embodiments, the alkyl contains from one to five carbon atoms (i.e., C1-C5 alkyl). In other embodiments, the alkyl contains from one to four carbon atoms (i.e., C1-C4 alkyl). In other embodiments, the alkyl contains from one to three carbon atoms (i.e., C1-C3 alkyl). In other embodiments, the alkyl contains from 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 from five to fifteen carbon atoms (i.e., C5-C 15 (alkyl). In other embodiments, the alkyl contains from five to eight carbon atoms (i.e., C5-C8 alkyl). In other embodiments, the alkyl contains from two to five carbon atoms (i.e., C2-C5 alkyl). In other embodiments, the alkyl contains from three to five carbon atoms (i.e., C3-C5 alkyl). In certain embodiments, the alkyl 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.

[0086] The term "C x-y ", when used in combination with a chemical moiety such as alkyl, alkenyl or alkynyl, refers to a group containing from x to y carbon atoms in the chain. For example, the term "C 1-6 alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group, which includes straight-chain and branched-chain alkyl groups containing 1 to 6 carbon atoms. The term "–C x-y alkylene-" refers to a substituted or unsubstituted alkylene chain containing from x to y carbon atoms in the alkylene chain. For example, "–C 1-6 alkylene-" can be selected from methylene, ethylene, propylene, butylene, pentylene and hexylene, any of which is optionally substituted.

[0087] "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.

[0088] "Alkenyl" refers to a straight-chain or branched-chain hydrocarbon chain group consisting only of carbon atoms and hydrogen atoms, containing at least one carbon-carbon double bond, and preferably having from 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 rest 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.

[0089] "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 rest of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc.

[0090] The term "C x-y alkenyl" and "C x-y alkynyl" refer to substituted or unsubstituted unsaturated aliphatic groups, whose length and possible substituents are similar to those of 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 containing 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 contain 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 containing 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 contain one triple bond or more than one triple bond in the alkynylene chain.

[0091] "Alkylene" or "alkylene chain" refers to a divalent hydrocarbon chain that connects the remainder of the molecule to a group, consists only of carbon and hydrogen, contains no unsaturation, and preferably has a straight or branched chain with 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 group by a single bond. The attachment points of the alkylene chain to the remainder of the molecule and to the group can be 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., C1-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).

[0092] "Alkenylene" or "alkenylene chain" refers to a divalent hydrocarbon chain that connects the remainder of the molecule to a group, consists only of carbon and hydrogen, contains at least one carbon-carbon double bond, and preferably has a straight or branched chain with two to twelve carbon atoms. The alkenylene chain is attached to the remainder of the molecule by a single bond and to the group by a single bond. The attachment points of the alkenylene chain to the remainder of the molecule and to the group can be any two carbons 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).

[0093] "Alkynylene" or "alkynylene chain" means a divalent hydrocarbon chain consisting only of carbon and hydrogen, containing at least one carbon-carbon triple bond, and preferably having a straight or branched chain of two to twelve carbon atoms, which connects the remainder of the molecule to a group. The alkynylene chain is attached to the remainder of the molecule by a single bond and to the group by a single bond. The attachment points of the alkynylene chain to the remainder of the molecule and to the group can be any two carbons 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).

[0094] "Aryl" means a group derived from an aromatic monocyclic or polycyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. The aromatic monocyclic or polycyclic hydrocarbon ring system contains only hydrogen and carbon and has five to eighteen carbon atoms, with at least one ring in the ring system being an aromatic ring, i.e., the ring system contains a cyclic, delocalized (4n + 2)π electron system that conforms to Hückel theory. Ring systems from which aryl is derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetrahydronaphthalene, and naphthalene.

[0095] "Aralkyl" means a group of the formula -R c -aryl, where R c is an alkylene chain as defined above, such as, for example, methylene, ethylene, etc.

[0096] "Arenylene" means a group of the formula -R d -aryl, where R d is an alkenylene chain as defined above. "Arylalkynyl" means a group of the formula -R e -aryl, where R e is an alkynylene chain as defined above.

[0097] "Carbocycle" refers to a saturated ring, an unsaturated ring or an aromatic ring, wherein each atom in the ring is carbon. The carbocycle 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 in the bicyclic carbocycle 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). Any combination of saturated bicyclic, unsaturated bicyclic, and aromatic bicyclic rings is included in the definition of carbocycle where valence allows. Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl. The bicyclic carbocycle can be a fused system, a bridged system, or a spiro system. In some cases, the spiro carbocycle has at least two molecular rings and only one common atom.

[0098] "Carbocyclene" refers to a divalent carbocycle that attaches the remainder of the molecule to a group.

[0099] The term "unsaturated carbocycle" refers to a carbocycle that has at least one degree of unsaturation and does not include an aromatic carbocycle. Examples of unsaturated carbocycles include cyclohexadiene, cyclohexene, and cyclopentene.

[0100] "Cycloalkyl" refers to a fully saturated monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms, which includes fused ring or bridged ring systems 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. Polycyclic cycloalkyls include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, 7,7-dimethylbicyclo[2.2.1]heptanyl, etc.

[0101] "Cycloalkenyl" refers to an unsaturated non-aromatic monocyclic or polycyclic hydrocarbon group consisting only of carbon and hydrogen atoms, which includes fused ring or bridged ring systems, preferably has three to twelve carbon atoms, and contains at least one double bond. In certain embodiments, the cycloalkenyl contains three to ten carbon atoms. In other embodiments, the cycloalkenyl contains five to seven carbon atoms. The cycloalkenyl can be attached to the remainder of the molecule by a single bond. Examples of monocyclic cycloalkenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.

[0102] "Cycloalkylalkyl" refers to a group of the formula -R c -cycloalkyl, where R c is an alkylene chain as described above.

[0103] "Cycloalkylalkoxy" refers to a group of the formula -O-Rc -cycloalkyl group bonded through an oxygen atom, wherein R c is an alkylene chain as described above.

[0104] "Halo" or "halogen" refers to halogen substituents such as bromo, chloro, fluoro, and iodo substituents.

[0105] As used herein, the term "haloalkyl" or "haloalkane" refers to an alkyl group as defined above that is substituted with one or more halogen groups, for example, trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. In some embodiments, the alkyl portion of the fluoroalkyl group is optionally further substituted. Examples of halogen-substituted alkanes ("haloalkanes") include methyl halides (e.g., methyl chloride, methyl bromide, methyl fluoride, methyl iodide), dihalomethanes and trihalomethanes (e.g., trichloromethane, tribromomethane, trifluoromethane, triiodomethane), 1-haloethane, 2-haloethane, 1,2-dihaloethane, 1-halopropane, 2-halopropane, 3-halopropane, 1,2-dihalopropane, 1,3-dihalopropane, 2,3-dihalopropane, 1,2,3-trihalopropane, and any other suitable combination of an alkane (or substituted alkane) and a halogen (e.g., Cl, Br, F, I, etc.). When an alkyl group is substituted with more than one halogen group, each halogen can be independently selected, for example, 1-chloro, 2-fluoroethane.

[0106] "Fluoroalkyl" refers to an alkyl group as defined above substituted with one or more fluoro groups, for example, trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like.

[0107] "Aminoalkyl" refers to an alkyl group as defined above substituted with one or more amine groups, for example, propane-2-amine, butane-1,2-diamine, pentane-1,2,4-triamine, and the like.

[0108] "Hydroxyalkyl" refers to an alkyl group as defined above substituted with one or more hydroxy groups, for example, 1-propanol, 1,4-butanediol, 1,2,4-pentanetriol, and the like.

[0109] "Alkoxyalkyl" refers to an alkyl group as defined above substituted with one or more alkoxy groups, for example, methoxymethane, 1,3-dimethoxybutane, 1-methoxypropane, 2-ethoxypentane, and the like.

[0110] As used herein, "cyanoalkyl" refers to an alkyl group as defined above substituted with one or more cyano groups, for example, acetonitrile, 2-ethyl-3-methylsuccinonitrile, butyronitrile, and the like.

[0111] "Heterocycle" means 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 saturated, unsaturated, and aromatic rings. Bicyclic heterocycles can be fused, bridged, or spiro ring systems. In some cases, a spiro heterocycle has at least two molecular rings and only one common atom. A spiro heterocycle contains at least one heteroatom.

[0112] "Heterocyclene" means a divalent heterocycle that attaches the remainder of the molecule to a group.

[0113] "Heteroaryl" or "aromatic heterocycle" means 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. The heteroaromatic rings as used herein can be selected from monocyclic or bicyclic and fused or bridged ring systems, where at least one ring in the ring system is an aromatic ring, i.e., it contains a cyclic, delocalized (4n + 2)π electron system that complies with Hückel's theory. The heteroatoms in the heteroaryl can be optionally oxidized. One or more nitrogen atoms (if present) can be optionally quaternized. Where valence allows, the heteroaryl can be attached to the remainder of the molecule through any atom of the heteroaryl, such as a carbon atom or a nitrogen atom of the heteroaryl. Examples of heteroaryl include, but are not limited to, pyridine, pyrimidine, oxazole, furan, pyran, thiophene, isoxazole, benzimidazole, benzothiazole, and imidazopyridine.

[0114] "X-membered heteroaryl" refers to the number of atoms within the ring, i.e., X. For example, a 5-membered heteroaromatic ring or a 5-membered aromatic heterocycle has 5 atoms within the ring, such as triazole, oxazole, thiophene, etc.

[0115] The term "unsaturated heterocycle" means a heterocycle that has at least one degree of unsaturation and does not include aromatic heterocycles. Examples of unsaturated heterocycles include dihydropyrrole, dihydrofuran, oxazoline, pyrazoline, and dihydropyridine. The heterocycle can be optionally substituted with one or more substituents, such as those described herein.

[0116] The term "substituted" means a moiety having a substituent that replaces a hydrogen on one or more carbon atoms or replaceable heteroatoms (such as NH) in the structure. It should be understood that "substituted" or "substituted with" includes the following implicit conditions: such substitution complies with the allowed valences of the atom being substituted and the substituent, and the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformations such as rearrangement, cyclization, elimination, etc. In certain embodiments, substituted means a moiety having a substituent that replaces two hydrogens on the same carbon atom, such as replacing two hydrogens on a single carbon atom with an oxo, imino, or thio group.

[0117] 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 that satisfy the valence of the heteroatom in the organic compounds described herein. 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 (=N-H), oxime (=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 by alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo(=O), thioxo(=S), cyano(-CN), nitro(-NO2), imino(=N-H), oxime(=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) substitution; where each R a is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, where each R aOptionally substituted by alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo(=O), thioxo(=S), cyano(-CN), nitro(-NO2), imino(=N-H), oxime(=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); where each R b is independently selected from a direct bond or a straight or branched alkylene, alkenylene or alkynylene chain, and each R c is a straight or branched alkylene, alkenylene or alkynylene chain.

[0118] As used in the specification and claims, unless the context clearly dictates otherwise, the singular forms "a", "an" and "the" include plural referents.

[0119] 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.

[0120] As used herein, the phrases "parenteral administration" and "administered parenterally" refer to a mode of administration other than enteral and topical administration and typically by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0121] As used herein, the phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that, within the scope of reasonable medical judgment, are suitable for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications and are commensurate with a reasonable benefit / risk ratio.

[0122] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refers to 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 used in pharmaceutical formulations.

[0123] In certain embodiments, the terms "prevent" or "preventing" in relation to a disease or disorder may refer to a compound that reduces the incidence of the disorder or condition in a treated sample relative to an untreated control sample in a statistical sample, or a compound that delays the onset or reduces the severity of one or more symptoms of the disorder or condition relative to an untreated control sample.

[0124] As used herein, the terms "treat", "treating", or "treatment" may include alleviating, reducing, or ameliorating the symptoms of a disease or disorder, preventing additional symptoms, ameliorating or preventing the underlying cause of the symptoms, inhibiting the disease or disorder, e.g., arresting the development of the disease or disorder, relieving the disease or disorder, causing regression of the disease or disorder, alleviating the condition caused by the disease or disorder, or prophylactically and / or therapeutically halting the symptoms of the disease or disorder.

[0125] 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 to which the ligand binds, including but not limited to protein hormones, lectins, growth factors, antibodies, or others capable of binding 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.

[0126] 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.

[0127] The term "linker" refers to a chemical moiety capable of joining two different chemical moieties to each other. The linker can include a spacer and an amino acid. The linker can be a cleavable linker, which 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).

[0128] The term "antibody" refers to a whole antibody and any antigen-binding fragment thereof (i.e., "antigen-binding portion") or single-chain variant. A whole antibody is a protein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain comprises a heavy-chain variable region (VH) and a heavy-chain constant region, which constant region comprises three domains: CH1, CH2, and CH3. Each light chain comprises a light-chain variable region (VL or Vk) and a light-chain constant region, which constant region comprises a single domain: CL. The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), interspersed with more conserved framework regions (FRs). Each VH and VL contains three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions contain the binding domains that interact with an antigen. The constant regions can mediate the binding of the antibody to host tissues or factors, which host tissues or factors include 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" antigen X. The antibody can be chimeric, humanized, or preferably human. The heavy-chain constant region can be engineered to affect the type or degree of glycosylation, to extend the antibody half-life, to enhance or reduce the interaction with effector cells or the complement system, or to modulate some other property. The engineering can be achieved by replacing, adding, or deleting one or more amino acids or by replacing one domain with a domain from another immunoglobulin type or a combination of the foregoing.

[0129] The terms "antigen-binding fragment" and "antigen-binding portion" of an antibody (or simply "antibody portion" or "antibody fragment") refer to one or more fragments of an antibody that retain the ability to specifically bind 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 fragments, monovalent fragments consisting of the VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, divalent fragments comprising two Fab fragments joined by a disulfide bridge at the hinge region; (iii) Fab' fragments, which are essentially 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 fragments consisting of the VH and CH1 domains; (v) Fv fragments consisting of the VL and VH domains of a single arm of an antibody; (vi) dAb fragments (Ward et al., (1989) Nature 341:544-546), which consist of a VH domain; (vii) isolated complementarity-determining regions (CDRs); and (viii) nanobodies, heavy chain variable regions 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 an Fv fragment are encoded by different genes, they can be joined using recombinant methods by a synthetic linker that enables them to be a single protein chain, in which the VL and VH regions pair to form a monovalent molecule (called 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 included within the term "antigen-binding portion" of an antibody.

[0130] The term "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigen specificities (e.g., an isolated antibody that specifically binds antigen X is substantially free of antibodies that specifically bind antigens other than antigen X). However, an isolated antibody that specifically binds antigen X may cross-react with other antigens, such as antigen X molecules from other species. In certain embodiments, an isolated antibody specifically binds a human antigen X and does not cross-react with other (non-human) antigen X antigens. In addition, an isolated antibody may be substantially free of other cellular materials and / or chemicals.

[0131] The term "monoclonal antibody" or "monoclonal antibody composition" refers to a preparation of antibody molecules of a single molecular composition that exhibit a single binding specificity and affinity for a particular epitope.

[0132] The term "human antibody" refers to an antibody having a variable region in which the framework region and the CDR regions (and the constant region, if present) are all derived from human germline immunoglobulin sequences. A human antibody can include post-translational modifications, including natural or synthetic modifications. A human antibody can 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.

[0133] The term "human monoclonal antibody" refers to an antibody that exhibits a single binding specificity and has a variable region in which the framework region and the CDR regions are both derived from human germline immunoglobulin sequences. In one embodiment, a human monoclonal antibody is produced by a hybridoma that comprises B cells obtained from a transgenic non-human animal (e.g., a transgenic mouse) having a genome that comprises a human heavy chain transgene and a light chain transgene that are fused to immortalized cells.

[0134] The compounds of the present disclosure

[0135] The following is a discussion of the compounds and their salts that can be used in the methods of the present disclosure.

[0136] In one aspect, the present disclosure provides a compound of formula (I):

[0137]

[0138] or a pharmaceutically acceptable salt thereof, wherein;

[0139] L 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 ;

[0140] L 2 is selected from C 1-6 alkylene;

[0141] L 2B is selected from (NR 4 ) tC(O)O-CH2-phenyl, wherein the phenyl group is optionally substituted by one or more R 5 replace;

[0142] L 2C Selected from C(O)O-CH2-phenyl, wherein the phenyl group is optionally replaced by one or more R 6 replace;

[0143] L 3 is selected from the group consisting of 1 to 7 amino acid residues;

[0144] L 4 Selected from optionally substituted C 1-6 Alkylene, wherein the C 1-6 The alkylene group is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 Haloalkyl, -OC 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Substitution of alkynyl groups by substituents;

[0145] L 5 Selected from (O-CH2-CH2-) q -(NR 3 ) s ;

[0146] L 6 Selected from optionally substituted C 1-6 Alkylene, wherein the C 1-6 The alkylene group is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 Alkyl, -C 1-10 Haloalkyl, -OC 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Substitution of alkynyl groups by substituents;

[0147] L 7 Selected from C 5-6 carbon ring;

[0148] R 1 Selected from -O- and -NR 7 -;

[0149] R 2 is selected from an optionally substituted 5- to 6-membered heterocyclic ring, wherein the 5- to 6-membered heterocyclic ring 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, -OC 1-10Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl;

[0150] R 3 Selected from hydrogen and C 1-6 Alkyl;

[0151] R 4 Selected from hydrogen and optionally substituted by one or more SO2C 1-6 Alkyl-substituted C 1-6 Alkyl;

[0152] Each R 5 Independently selected from sugars;

[0153] Each R 6 Independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 Halogenoalkyl, -O-C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl;

[0154] R 7 Selected from hydrogen and C 1-6 Alkyl;

[0155] R 8 Selected from hydrogen and hydroxyl;

[0156] R 9 Selected from hydrogen and halogen, wherein R 8 Or R 9 At least one of which is hydrogen;

[0157] m is selected from 0 and 1;

[0158] n is selected from 0 and 1;

[0159] p is selected from 0 and 1;

[0160] q is selected from 0 to 8;

[0161] s is selected from 0 and 1;

[0162] t is selected from 0 and 1;

[0163] x is selected from 0 and 1;

[0164] y is selected from 0 and 1; and

[0165] z is selected from 0 and 1.

[0166] In some embodiments, formula (I) is represented as

[0167]

[0168] Or a pharmaceutically acceptable salt thereof.

[0169] In some embodiments, formula (I) is represented as

[0170]

[0171] In some embodiments, for a compound or salt of formula (I), formula (I-A) or formula (I-B), L is L 2 -(L 2B ) z -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 . In some cases, L is L 2 -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 . In some cases, L is L 2 -L 3 -L 4 -L 5 -L 6 -R 2 . In some cases, L is L 2 -L 3 -L 4 -L 7 -R 2 . In some cases, the linker is L 2 -L 3 -L 4 -R 2 . In some cases, L is L 2 -L 2B -L 3 -L 4 -L 5 -R 2 . In some cases, L is L 2 -L 2B -L 3 -L 4 -R 2 . In some cases, L is L 2B -L 3 -L 4 -R 2 .

[0172] In some embodiments, for a compound or salt of formula (I), formula (I-A) or formula (I-B), L is (L 2B ) z -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 。In some cases, L is L 2B -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 。In some cases, L is L 2B -L 3 -L 4 -L 5 -L 6 -R 2 。In some cases, L is L 2B -L 3 -L 4 -L 5 -R 2 。In some cases, L is L 2B -L 3 -L 4 -R 2 。

[0173] In some embodiments, for a compound or salt of formula (I), formula (I-A) or formula (I-B), L is (L 2C ) y -L 3 -L 4 -(L 5 ) m -(L 6 ) n -(L 7 ) p -R 2 。In some cases, L is L 2C -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 。In some cases, L is L 2C -L 3 -L 4 -L 5 -L 6 -R2 。In some cases, L is L 2C -L 3 -L 4 -L 5 -R 2 。In some cases, L is L 2C -L 3 -L 4 -R 2 。In some cases, L is L 2C -L 3 -L 4 -L 7 -R 2 。In some cases, L is L 3 -L 4 -R 2 。

[0174] In some embodiments, for a compound or salt of formula (I), formula (I-A) or formula (I-B), 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.

[0175] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III) or formula (III-A), 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, L2B 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

[0176] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III), or formula (III-A), 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

[0177] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III), or formula (III-A), L 3 is selected from residues comprising 1 to 7 amino acids. In some cases, L 3 is selected from residues comprising 1 to 5 amino acids. In some cases, L 3 is selected from residues comprising 1 to 4 amino acids. In some cases, L 3Residues selected from those containing 1 to 3 amino acids. In some cases, L 3 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.

[0178] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III) or formula (III-A), L 3 is selected from residues containing natural and unnatural amino acids.

[0179] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III) or formula (III-A), L 3 is selected from residues containing natural amino acids. In some cases, L 3 is selected from residues containing α-amino acids. In some cases, L 3 is selected from residues containing β-amino acids.

[0180] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III) or formula (III-A), the amino acid is an unnatural amino acid.

[0181] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III) or formula (III-A), L 3 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, L 3is selected from the group consisting of alanine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, isoleucine, leucine, lysine, phenylalanine, serine, valine, citrulline, sarcosine, and β-alanine. 3 is selected from the group consisting of alanine, glycine, lysine, phenylalanine, serine, valine, citrulline, sarcosine, and β-alanine. 3 is selected from the group consisting of alanine, glycine, phenylalanine, valine, citrulline, and β-alanine. 3 is selected from the group consisting of alanine, valine, and citrulline. 3 is selected from glycine and phenylalanine. In some cases, L 3 is selected from the group consisting of glycine, phenylalanine, and sarcosine. 3 Selected from sarcosine.

[0182] In some embodiments, for the compound or salt of Formula (I), Formula (IA), Formula (IB), Formula (II), Formula (III), or Formula (III-A), L 3 Contains 4 amino acids. In some cases, L 3 Contains 2 amino acids. In some cases, L 3 Contains 1 amino acid. In some cases, L 3 Contains at least two different amino acids. In some cases, L 3 yes In some cases, L 3 yes In some cases, L 3 yes In some cases, L 3 yes

[0183] In some embodiments, for the compound or salt of Formula (I), Formula (IA), Formula (IB), Formula (II), Formula (III), or Formula (III-A), L 4 Selected from optionally substituted C 1-6 Alkylene, wherein the C 1-6 The alkylene group is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 Haloalkyl, -OC 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 In some cases, L 4 Selected from optionally substituted C 1-6 Alkylene, wherein the C 1-6The alkylene group is optionally substituted with 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 with 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-.

[0184] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III), or formula (III-A), 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 certain 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 5 is -(O-CH2-CH2-)7-NH-. In some cases, L 5is -(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.

[0185] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III) or formula (III-A), L 6 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. In some cases, L 6 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 -OH, -NH2 and oxo. In some cases, L 6 is selected from optionally substituted C 1-6 alkylene, wherein the 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-.

[0186] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III) or formula (III-A), L 7 is selected from C 5-6 carbocycle. In some cases, L 7 is selected from C 5-6 subcarbocycle. In some cases, L 7 is selected from C6 carbocycle. In some cases, L 7 is phenyl. In some cases, L 7 is phenylene. In some cases, L 7 is cyclohexyl. In some cases, L7 Yes In some cases, L 7 Yes

[0187] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II) or formula (III), 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 1-10 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 heteroalicyclic ring, wherein the 5- to 6-membered heteroalicyclic ring 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. 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 by 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 heteroalicyclic ring, wherein the 5- to 6-membered heteroalicyclic ring is optionally substituted by 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 by one or more substituents independently selected from oxo. In some cases, R 2 is selected from an optionally substituted 5-membered heteroalicyclic ring, wherein the 5-membered heteroalicyclic ring is optionally substituted by 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 Yes In some cases, R 2 Yes wherein Lg is a ligand. 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.

[0188] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III), or formula (III-A), L 2 -L 3 -L 4 -R 2 is selected from

[0189] In some cases, L 2 -L 3 -L 4

[0190]

[0191] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), formula (III), or formula (III-A), L 2 -L 3 -L 4 is In some cases, L 2 -L 3 is

[0192] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II), or formula (III), 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 In some cases, L 2 -L 3 -L 4 -L 5 is In some cases, L 2 -L 3 -L4 -L 5 is

[0193] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II) or formula (III), 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

[0194] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II) or formula (III), L is

[0195]

[0196] In some embodiments, for a compound or salt of formula (I), formula (I-A), formula (I-B), formula (II) or formula (III), 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 ; wherein L 2C is selected from

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

[0198]

[0199] or a pharmaceutically acceptable salt thereof, wherein Lg is a ligand.

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

[0201]

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

[0203] Lg is the ligand;

[0204] L 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 ;

[0205] L 2 Selected from C 1-6 alkylene;

[0206] L 2B Selected from (NR 4 ) t C(O)OC 1-6 Alkylene-phenyl, wherein the phenyl group is optionally replaced by one or more R 5 replace;

[0207] L 2C Selected from C(O)O-CH2-phenyl, wherein the phenyl group is optionally substituted with halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 Haloalkyl, -OC 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl substitution;

[0208] L 3 is selected from the group consisting of 1 to 7 amino acid residues;

[0209] L 4 Selected from optionally substituted C 1-6 Alkylene, wherein the C 1-6 The alkylene group is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, -C1-10 Haloalkyl, -OC 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Substitution of alkynyl groups by substituents;

[0210] L 5 Selected from (O-CH2-CH2-) q -(NR 3 ) s ;

[0211] L 6 Selected from optionally substituted C 1-6 Alkylene, wherein the C 1-6 The alkylene group is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 Alkyl, -C 1-10 Haloalkyl, -OC 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Substitution of alkynyl groups by substituents;

[0212] L 7 Selected from C 5-6 carbon ring;

[0213] R 1 Selected from -O- and -NR 7 -;

[0214] R 2 is selected from an optionally substituted 5- to 6-membered heterocyclic ring, wherein the 5- to 6-membered heterocyclic ring is optionally substituted by one or more independently selected from halogen, -OH, -CN, -NO2, -NH2, oxo, C 1-10 Alkyl, -C 1-10 Haloalkyl, -OC 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Substitution of alkynyl groups by substituents;

[0215] R 3 Selected from hydrogen and C 1-6 alkyl;

[0216] R 4 selected from hydrogen and optionally one or more SO2C 1-6 Alkyl substituted C 1-6 alkyl;

[0217] Each R 5 independently selected from sugars;

[0218] R 7 Selected from hydrogen and C 1-6 alkyl;

[0219] R 8 is selected from hydrogen and hydroxyl;

[0220] R 9 is selected from hydrogen and halogen, wherein R 8 or R 9 at least one of which is hydrogen;

[0221] m is selected from 0 and 1;

[0222] n is selected from 0 and 1;

[0223] p is selected from 0 and 1;

[0224] q is selected from 0 to 8;

[0225] s is selected from 0 and 1;

[0226] t is selected from 0 and 1;

[0227] x is selected from 0 and 1;

[0228] y is selected from 0 and 1; and

[0229] z is selected from 0 and 1.

[0230] In some embodiments, formula (III) is represented as

[0231] or a pharmaceutically acceptable salt thereof.

[0232] In one aspect, the present disclosure provides a compound of formula (IV-A)

[0233]

[0234] or a pharmaceutically acceptable salt thereof.

[0235] In one aspect, the present disclosure provides a compound of formula (IV-B)

[0236]

[0237] or a pharmaceutically acceptable salt thereof.

[0238] In one aspect, the present disclosure provides a compound of formula (IV-C)

[0239]

[0240] or a pharmaceutically acceptable salt thereof.

[0241] In one aspect, the present disclosure provides a compound of formula (IV-D)

[0242]

[0243] or a pharmaceutically acceptable salt thereof.

[0244] This disclosure includes salts of the compounds described herein, particularly pharmaceutically acceptable salts. Compounds of the invention having sufficient acidity, sufficient basicity, or both functional groups can react with a variety of inorganic bases and with any of a variety of inorganic and organic acids to form salts. Alternatively, inherently charged compounds (such as those having a quaternary nitrogen) can form salts with appropriate counterions (such as halides, such as bromide, chloride, or fluoride, particularly bromide).

[0245] Chemical entities having a carbon-carbon double bond or a carbon-nitrogen double bond can exist in the Z- or E-form (or cis- or trans-form). In addition, some chemical entities can exist in various tautomeric forms. Unless otherwise indicated, the compounds described herein are also intended to include all Z-, E-, and tautomeric forms.

[0246] A "tautomer" refers to a molecule in which the transfer of a proton from one atom of the molecule to another atom of the same molecule is possible. In certain embodiments, the compounds presented herein exist in tautomeric forms. Where tautomerism can occur, there will be a chemical equilibrium of the tautomers. The exact proportions of the tautomers depend on several factors, including physical state, temperature, solvent, and pH. Some examples of tautomeric equilibria include:

[0247]

[0248] In some embodiments, the compounds disclosed herein are used in different enriched isotope forms, e.g., enriched 2 H, 3 H, 11 C, 13 C, and / or 14 C content. In one 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 increasing the duration of drug action.

[0249] Unless otherwise indicated, the compounds described herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure of the invention are within the scope of this disclosure except where hydrogen is replaced by deuterium or tritium, or carbon is replaced by 13 C- or 14 C-enriched carbon.

[0250] The compounds of the present disclosure optionally contain unnatural proportions of atomic isotopes on one or more atoms that make up such compounds. For example, the compounds may be isotopically labeled, such as, for example, 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 contemplated. All isotopic variants of the compounds of the invention, whether radioactive or not, are encompassed within the scope of the invention.

[0251] In certain embodiments, some or all of the 1 H atoms of the compounds disclosed herein are replaced by 2 H atoms. Methods for synthesizing deuterium-containing compounds are known in the art and include, by way of non-limiting example only, the following synthetic methods.

[0252] Deuterium-substituted compounds are synthesized using a variety of methods such as those described in: 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, Anthony S. Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.

[0253] Deuterated starting materials are readily available and are subjected to the synthetic methods described herein to provide 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.

[0254] 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, unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, and mixtures thereof.

[0255] In some cases, the compounds described herein may exist as diastereoisomers, enantiomers or other stereoisomeric forms. In the absence of designation of absolute stereochemistry, the compounds presented herein include all diastereoisomeric forms, enantiomeric forms 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, incorporated herein by reference for this disclosure). Stereoisomers can also be obtained by stereoselective synthesis.

[0256] The methods and compositions described herein include the use of amorphous forms as well as crystalline forms (also referred to as polymorphs). The compounds described herein may 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 may exist in unsolvated forms as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc. The solvated forms of the compounds presented herein are also considered to be disclosed herein.

[0257] In certain embodiments, the compound or its salt may be a prodrug, for example, where a hydroxyl group in the parent compound is presented in the form of an ester or carbonate, or a carboxylic acid in the parent compound is presented in the form of an ester. The term “prodrug” is intended to cover compounds that are converted into the agents of the present disclosure under physiological conditions. One method of preparing a prodrug is to include one or more selected moieties that are hydrolyzed 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 in the host animal. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids and esters of phosphonic acids) are preferred prodrugs of the present disclosure.

[0258] Prodrug forms of the compounds described herein, where the prodrug is metabolized in vivo to produce the compounds described herein, are included within the scope of the claims. In some cases, some of the compounds described herein may be prodrugs of another derivative or active compound.

[0259] 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 drug is not. Prodrugs can help enhance the cellular permeability of a compound relative to the parent drug. Prodrugs can also have better solubility in a pharmaceutical composition than the parent drug. Prodrugs can be designed as reversible drug derivatives that serve as modifiers to enhance the transport of a drug to a specific tissue site or increase the residence of the drug within a cell.

[0260] 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, the A.C.S. Symposium Series Vol. 14; and Edward B. Roche, Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, all incorporated herein by reference for such disclosure). According to another embodiment, the present disclosure provides methods for preparing the compounds defined above. The compounds can be synthesized using conventional techniques. Advantageously, these compounds can be conveniently synthesized from readily available starting materials.

[0261] The synthetic chemical transformations and methods for synthesizing the compounds described herein are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations (1989); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2nd 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).

[0262] Ligand

[0263] In some embodiments, for the compounds or salts of formula (I), formula (I-A), formula (I-B), formula (II), formula (III), or formula (III-A), the ligand is selected from an antibody or an antigen-binding fragment thereof. In some cases, the ligand is selected from chimeric antibodies, humanized antibodies, and human antibodies.

[0264] 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 target cell is a cancer tissue or cancer cell, and the antigen or receptor is a tumor-associated antigen, i.e., an antigen that is uniquely expressed by cancerous cells or overexpressed by cancer cells 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. In some cases, the ligand is selected from UC-961, PTK-7, trastuzumab, brentuximab, loncastuximab, rosopatamab, rituximab, pinatuzumab, polatuzumab, and naratuximab. In some cases, the ligand is selected from trastuzumab, brentuximab, loncastuximab, rosopatamab, rituximab, pinatuzumab, polatuzumab, and naratuximab. In some cases, the ligand is trastuzumab.

[0265] 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 Korman et al., U.S. Patent No. 8,609,816 B2 (2013; B7H4, also known as O8E; especially antibodies 2A7, 1G11, and 2F9); Rao-Naik et al., U.S. Patent No. 8,097,703 B2 (2012; CD19; especially antibodies 5G7, 13F1, 46E8, 21D4, 21D4a, 47G4, 27F3, and 3C10); King et al., U.S. Patent No. 8,481,683 B2 (2013; CD22; especially antibodies 12C5, 19A3, 16F7, and 23C6); Keler et al., U.S. Patent No. 7,387,776 B2 (2008; CD30; especially antibodies 5F11, 2H9, and 17G1); Terrett et al., U.S. Patent No. 8,124,738 B2 (2012; CD70; especially antibodies 2H5, 10B4, 8B5, 18E7, and 69A7); Korman et al., U.S. Patent No. 6,984,720 B1 (2006; CTLA-4; especially antibodies 10D1, 4B6, and 1E2); Vistica et al., U.S. Patent No. 8,383,118 B2 (2013, fucosyl-GM1, especially antibodies 5B1, 5B1a, 7D4, 7E4, 13B8, and 18D5); Korman et al., U.S. Patent No. 8,008,449 B2 (2011; PD-1; especially antibodies 17D8, 2D3, 4H1, 5C4, 4A11, 7D3, and 5F4); Huang et al., US2009 / 0297438 A1 (2009; PSMA, especially antibodies 1C3, 2A10, 2F5, 2C6); Cardarelli et al., U.S. Patent No. 7,875,278 B2 (2011; PSMA; especially antibodies 4A3, 7F12, 8C12, 8A11, 16F9, 2A10, 2C6, 2F5, and 1C3); Terrett et al., U.S. Patent No. 8,222,375 B2 (2012; PTK7; especially antibodies 3G8, 4D5, 12C6, 12C6a, and 7C8);Terrett et al., U.S. Patent No. 8,680,247 B2 (2014; glypican-3; specifically antibodies 4A6, 11E7, and 16D10); Harkins et al., U.S. Patent No. 7,335,748 B2 (2008; RG1; specifically antibodies A, B, C, and D); Terrett et al., U.S. Patent No. 8,268,970 B2 (2012; mesothelin; specifically antibodies 3C10, 6A4, and 7B1); Xu et al., US2010 / 0092484 A1 (2010; CD44; specifically antibodies 14G9.B8.B4, 2D1.A3.D12, and 1A9.A6.B9); Deshpande et al., U.S. Patent No. 8,258,266 B2 (2012; IP10; specifically 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; specifically antibodies F7, F9, D1, and E2); and Korman et al., U.S. Patent No. 7,943,743 B2 (2011; PD-L1; specifically antibodies 3G10, 12A4, 10A5, 5F8, 10H10, 1B12, 7H1, 11E6, 12B7, and 13G4); the disclosures of which are incorporated herein by reference.

[0266] 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.

[0267] In some embodiments, any of several different reactive groups on the ligand can serve as a conjugation site, including the ε-amino group in lysine residues, side-chain carbohydrate moieties, carboxylic acid groups, disulfide groups, and thiol groups. 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 & Therapeutics 83 (1999), 67-123, the disclosures of which are incorporated herein by reference.

[0268] In some embodiments, the ligand is conjugated through a lysine ε-amino group. Most antibodies have multiple lysine ε-amino groups, which can be conjugated using techniques known in the art via amide, urea, thiourea, or carbamate linkages. 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 may lead to the neutralization of protonated ε-amino groups that are important for maintaining the native conformation of the antibody, or may occur on lysines that are near or in the antigen-binding site, neither of which is desirable.

[0269] In some embodiments, since many antibodies are glycosylated, the ligand can be conjugated through a carbohydrate side chain. The carbohydrate side chain can be oxidized with periodic acid to generate an aldehyde group, which can then react with an amine to form an imine group, such as in a semicarbazone, oxime, or hydrazone. If desired, the imine group can be converted to a more stable amine group by reduction with sodium cyanoborohydride. For additional disclosures regarding conjugation through carbohydrate side chains, see, e.g., Rodwell et al., Proc. Nat'l Acad. Sci. USA 83, 2632-2636 (1986); the disclosure of which is incorporated herein by reference. As with lysine ε-amino groups, there are concerns regarding the reproducibility of the conjugation site location and stoichiometry.

[0270] In some embodiments, the ligand can be conjugated through 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.

[0271] In some embodiments, antibodies can be conjugated by bridging the disulfide groups of cysteine residues on the antibody and sulfur on another part of the conjugate or compound. Some antibodies lack free thiol (mercapto) groups but have disulfide groups, such as in the hinge region. In such cases, free thiol groups can be generated by reducing the native disulfide groups. The thiol groups so 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), 7,78 - 784 (2003); the disclosures of which are incorporated herein by reference. A variety of methods are known for introducing free thiol groups into antibodies without disrupting native disulfide bonds, and the methods can be practiced using the ligands of the present invention. Depending on the method employed, it may be possible to introduce a predictable number of free thiols 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). The preferred method for introducing free cysteine is taught by Liu et al., WO 2009 / 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 far from the antigen-binding site. The disclosures of the documents cited in this paragraph are incorporated herein by reference.

[0272] In some embodiments, the lysine ε-amino group can be modified with a reagent such as iminothiolane or N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) to convert the ε-amino group to a thiol or disulfide group, thereby generating a cysteine surrogate.

[0273] Linker

[0274] Compounds and salts of formula (I), formula (I-A), formula (I-B), formula (II), formula (III), or formula (III-A) may comprise 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 is also bound 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 may 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 a target, which can be a cognate binding partner such as an antigen. The conjugate may comprise multiple linkers, each having one or more compounds attached thereto. These linkers can be the same linker or different linkers.

[0275] In some embodiments, the linker can be short, flexible, rigid, cleavable, non-cleavable, hydrophilic, or hydrophobic. The linker can comprise segments having different properties, such as flexible segments or rigid segments. The linker can have chemical stability in the extracellular environment, e.g., in the bloodstream, or can comprise labile or selectively stable linkages. The linker can comprise linkages designed to specifically or non-specifically cleave and / or immolate or otherwise break intracellularly. Cleavable linkers can be sensitive to enzymes. Cleavable linkers can be cleaved by enzymes (e.g., proteases). Cleavable linkers can comprise valine-citrulline linkers or valine-alanine peptides. Linkers containing valine-citrulline or valine-alanine can comprise maleimide or succinimide groups.

[0276] In some embodiments, the non-cleavable linker can be protease-insensitive. The non-cleavable linker can be a maleimidohexanoyl linker. The maleimidohexanoyl linker can comprise N-maleimidomethylcyclohexane-1-carboxylate. The maleimidohexanoyl linker can comprise a succinimide group. The maleimidohexanoyl linker can comprise a pentafluorophenyl group. The linker can be a combination of maleimidohexanoyl and one or more polyethylene glycol molecules. The linker can be a maleimide-PEG4 linker. The linker can be a combination of a maleimidohexanoyl linker containing a succinimide group and one or more polyethylene glycol molecules. The linker can be a combination of a maleimidohexanoyl linker containing a pentafluorophenyl group and one or more polyethylene glycol molecules. The linker can comprise a maleimide attached to a polyethylene glycol molecule, where the polyethylene glycol can make the linker more flexible or can be used to extend the linker. The linker can be a (maleimidohexanoyl)-(valine-citrulline)-(p-aminobenzyloxycarbonyl) linker. The linker can be a linker suitable for attachment to engineered cysteine (THIOMAB). The THIOMAB linker can be a (maleimidohexanoyl)-(valine-citrulline)-(p-aminobenzyloxycarbonyl)-linker.

[0277] In some embodiments, the linker can further comprise an alkylene, alkenylene, alkynylene, polyether, polyester, polyamide group, and polyamino acid, polypeptide, cleavable peptide or aminobenzyl carbamate. The linker can comprise a maleimide at one end and an N-hydroxysuccinimide ester at the other end. The linker can comprise an N-terminal amine acetylated lysine and a valine-citrulline cleavage site. The linker can be a linkage generated 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 comprise a reactive primary amine. The linker can be a sortase A (SortaseA) linker. The sortase A linker can be generated by fusing the LXPTG recognition motif with an N-terminal GGG motif by the sortase A enzyme, thereby regenerating the native amide bond. Thus, the generated linker can link the moiety attached to the LXPTG recognition motif with the moiety attached to the N-terminal GGG motif.

[0278] In some embodiments, a compound or salt of any of the formulas described herein is attached to an antibody construct via a linker (also referred to herein as L or linker). L as used herein can be selected from any of the linker moieties discussed herein. The linker connecting the compound or salt to the antibody construct in the conjugate can be short, long, hydrophobic, hydrophilic, flexible, or rigid, or can consist of segments that each independently have one or more of the above characteristics such that the linker can contain segments with different characteristics. The linker can be multivalent such that they covalently attach more than one compound or salt to a single site on the antibody construct; or monovalent such that they covalently attach a single compound or salt to a single site on the antibody construct.

[0279] 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 having from about 10 to about 300 atoms in the linker. In some embodiments, the linker can have from about 30 to about 400 atoms in the linker, such as from about 30 to about 300 atoms.

[0280] In some embodiments, the linker described herein can attach a compound or salt of any of the formulas described herein to a ligand (e.g., an antibody) via a covalent bond between the linker and the antibody construct and the compound. The linker can contain functional groups capable of covalently attaching to a ligand (e.g., an antibody).

[0281] 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.

[0282] 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 bonds. 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 chemical or enzymatically cleavable chemical bonds while the remainder of the linker can be non-cleavable.

[0283] In some embodiments, the linker can contain chemically labile groups such as hydrazone and / or disulfide groups. Linkers containing chemically labile groups can take advantage of the differential properties between plasma and some cytoplasmic compartments. Cleavable linkers can also include disulfide groups.

[0284] In some embodiments, acid-labile groups (such as hydrazone) can remain intact during systemic circulation in the neutral pH environment of blood (pH 7.3 - 7.5) and once the antibody construct benzazepine The compound conjugate can be internalized into the weakly acidic endosome (pH 5.0 - 6.5) and lysosome (pH 4.5 - 5.0) compartments of the cell, where hydrolysis can occur and release the benzazepine compound. This pH-dependent release mechanism may be related to the non-specific release of the drug. To increase the stability of the linker hydrazone group, the linker can be modified by chemical modification (such as substitution), and thus adjusted to achieve more efficient release in lysosomes while minimizing losses in circulation. The hydrazone-containing linker 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 linkers containing cis-aconityl. Cis-aconityl chemistry can use carboxylic acids juxtaposed to the amide bond to accelerate amide hydrolysis under acidic conditions.

[0285] 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 contain peptide regions that can serve as enzyme substrates. Peptide-based linkers can be more stable in plasma and the extracellular environment than chemically labile linkers. Peptide bonds can have good serum stability because lysosomal proteolytic enzymes may have very low activity in blood due to endogenous inhibitors and the unfavorable high pH of blood compared to lysosomes. The release of the compounds described herein from antibody-drug conjugates can occur due to the action of lysosomal proteases (such as cathepsins and plasmin). These proteases are 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 may have lower hydrophobicity compared to longer 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.

[0286] In some embodiments, the cleavable linker can include non-cleavable moieties or segments, and / or the cleavable segments or moieties can be included in an otherwise non-cleavable linker to make it cleavable.

[0287] In some embodiments, the linker may comprise an enzymatically cleavable peptide moiety. The peptide may be selected from natural amino acids, unnatural amino acids, or combinations thereof. In certain embodiments, the peptide may be selected from dipeptides, tripeptides, or tetrapeptides. In a specific embodiment, the dipeptide may comprise L-amino acids and be 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.

[0288] Pharmaceutical formulation

[0289] In certain embodiments, provided herein are compositions (also referred to herein as "pharmaceuticals") comprising a therapeutically effective amount of a compound or salt of any one of formula (I), formula (I-A), formula (I-B), formula (II), formula (III), formula (III-A), formula (IV-A), formula (IV-B), formula (IV-C), or formula (IV-D).

[0290] The pharmaceutical composition can be formulated using one or more physiologically acceptable carriers (including excipients and adjuvants), which assist in processing the agent into a pharmaceutically useful preparation. Suitable preparations depend on the chosen route of administration. An overview of pharmaceutical compositions can be 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).

[0291] 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 a non-human mammal. When administered to an animal (such as a human), the composition or agent is preferably administered in the form of a pharmaceutical composition that contains, for example, the agent 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 glycols, glycerol, oils (such as olive oil) or injectable organic esters). In a preferred embodiment, when such a pharmaceutical composition is used for human administration, particularly for invasive routes of administration (e.g., routes such as injection or implantation that bypass transport or diffusion through the epithelial barrier), the aqueous solution is pyrogen-free or substantially pyrogen-free. For example, excipients can be selected to achieve delayed release of the drug or to selectively target one or more cells, tissues, or organs. The pharmaceutical composition can be in unit dosage form, such as tablets, capsules, granules, lyophilized agents for reconstitution, powders, solutions, syrups, suppositories, injections, etc. The composition can also be present in a transdermal delivery system, such as a skin patch. The composition can also be present in a solution suitable for topical administration, such as eye drops.

[0292] Pharmaceutically acceptable excipients can include physiologically acceptable agents that serve, for example, to stabilize a compound such as a pharmaceutical agent, increase its solubility or increase its absorption. 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 can be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (formulation) can also be a liposome or other polymeric matrix, into which, for example, the compounds of the present invention can be incorporated. For example, liposomes containing phospholipids or other lipids are non-toxic, physiologically acceptable and metabolizable carriers that are relatively simple to manufacture and administer.

[0293] The pharmaceutical composition (formulation) can be administered to a subject by any of a variety of routes of administration, which include, for example, oral, such as in an infusion, tablet, capsule (including sprinkle capsules and gelatin capsules), bolus, powder, granule, paste for application to the tongue, in an aqueous or non-aqueous solution or suspension; transmucosal absorption through the oral mucosa, such as sublingual absorption; anal, rectal or vaginal, such as in a vaginal suppository, cream or foam; parenteral, including intramuscular, intravenous, subcutaneous or intrathecal, such as in a sterile solution or suspension; nasal; intraperitoneal; subcutaneous; transdermal, such as applied to the skin in the form of a patch; and topical, such as applied to the skin in the form of a cream, ointment or spray, or in the form of eye drops. The compound can also be formulated for inhalation. In certain embodiments, the compound can simply be dissolved or suspended in sterile water.

[0294] The pharmaceutical composition can be a sterile aqueous or non-aqueous solution, suspension or emulsion, such as a microemulsion. The excipients described herein are exemplary and in no way limiting. 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 that is effective to produce the desired therapeutic effect.

[0295] Typically, the therapeutic efficacy of a subject can be monitored using assays and methods suitable for the condition being treated, which assays will be familiar to those of ordinary skill 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 agent or metabolite in a biological fluid from the subject (e.g., blood, blood fraction such as serum, and / or urine), and / or other biological samples or biological tissues. Any method for detecting a pharmaceutical agent practiced in the art and described herein can be used to measure the level of the pharmaceutical agent or metabolite during treatment.

[0296] The dosage of the agents described herein for treating a disease or disorder may 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 body 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, which is determined by those skilled in the medical arts. In addition to the factors described herein and above related to the use of the agents for treating a disease or disorder, the appropriate duration and frequency of administration of the agents can also be determined or adjusted by factors such as the condition of the patient, 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 may depend on the body mass, body weight, or blood volume of the subject. It is generally preferred to use the minimum dosage sufficient to provide an effective treatment. The design and conduct of preclinical and clinical studies of the agents described herein (including when administered for prophylactic benefits) are entirely within the skill 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 may be different, such as lower than the dosage when any one of the agents is administered alone as a single-agent therapy. In certain specific embodiments, the two agents in the combination may act synergistically or additively, and either agent can be used in a lesser amount than when administered alone. The amount of the agent that can be administered daily can be, for example, between about 0.01 mg / kg body weight and 100 mg / kg body weight, such as between about 0.1 to 1 mg / kg body weight, between about 1 to 10 mg / kg body weight, between about 10 - 50 mg / kg body weight, between about 50 - 100 mg / kg body weight. In other embodiments, the amount of the agent that can be administered daily is between about 0.01 mg / kg body weight and 1000 mg / kg body weight, between about 100 - 500 mg / kg body weight, or between about 500 - 1000 mg / kg body weight. The optimal dosage per day or per course of treatment may vary depending on the disease or disorder being treated and may also vary depending on the route of administration and the treatment regimen.

[0297] The pharmaceutical composition comprising the agent can be formulated in a manner suitable for the delivery method using conventional practice techniques in the art. The composition can be in the form of a solid (e.g., tablet), capsule, semi-solid (e.g., gel), liquid, or gas (e.g., aerosol). In other embodiments, the pharmaceutical composition is administered by bolus infusion.

[0298] Pharmaceutically acceptable excipients are well known in the pharmaceutical art and are 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 type of excipient is selected based on the mode of administration and the chemical composition of the active ingredient. Alternatively, the compositions described herein can be formulated as lyophilizates. The compositions described herein can be lyophilized or otherwise formulated into a lyophilized product, and the medicament in the composition is dissolved and / or diluted upon administration using a solution of one or more suitable excipients. In other embodiments, the medicament can be encapsulated in liposomes using techniques known and practiced in the art. In certain specific embodiments, the medicament is not formulated in liposomes for use in a stent 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.

[0299] For example, a pharmaceutical composition for oral administration or for injection, infusion, subcutaneous delivery, intramuscular delivery, intraperitoneal delivery, or other methods can be in liquid form. The liquid pharmaceutical composition can contain, for example, one or more of the following: sterile diluents such as water, saline solution (preferably physiological saline), Ringer's solution, isotonic sodium chloride, non-volatile oils that can be used as solvents or suspending media, polyethylene glycol, glycerol, propylene glycol, or other solvents; antibacterial agents; antioxidants; chelating agents; buffers and agents for adjusting tonicity, such as sodium chloride or glucose. The parenteral composition can be encapsulated in an ampoule, a disposable syringe, or a multi-dose vial made of glass or plastic. Physiological saline is preferably used, and the injectable pharmaceutical composition is preferably sterile. In another embodiment, for treating ophthalmic conditions or diseases, the liquid pharmaceutical composition can be applied to the eye in the form of eye drops. The liquid pharmaceutical composition can be delivered orally.

[0300] For oral preparations, 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 if desired, can be used in combination with diluents, buffers, wetting agents, preservatives, colorants and flavorants. The agent can be formulated with a buffer to provide protection of the compound from the gastric environment and / or the low pH value of the enteric coating. The agent contained in the pharmaceutical composition can be formulated with a flavorant (e.g., in the form of a liquid, solid or semi-solid preparation) and / or with an enteric coating for oral delivery.

[0301] A pharmaceutical composition comprising any 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 by implantation into the desired target site. Sustained release formulations can contain the compound dispersed in a carrier matrix and / or 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 site of implantation, the rate of release and the expected duration, as well as the nature of the condition, disease or disorder to be treated or prevented.

[0302] In certain embodiments, the pharmaceutical composition comprising the 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. Preferably in the form of a controlled release or sustained release formulation, which is topically administered or directly injected into the skin near or within the area to be treated, such as intradermal or subcutaneous injection. 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.

[0303] A pharmaceutical composition comprising an agent can be formulated as an emulsion for topical application. The emulsion comprises a liquid distributed in a second liquid continuous phase. 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 comprise one or more surfactants, emulsifying agents, emulsion stabilizers, buffering agents, and other excipients. The oil phase can comprise 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.

[0304] Ointments and creams can be formulated, for example, with an aqueous or oily base to which a suitable thickening agent and / or gelling agent is added. Lotions can be formulated with an aqueous or oily base and generally will also contain one or more emulsifying agents, stabilizers, dispersing agents, suspending agents, thickening agents, or coloring agents. Liquid sprays can be delivered, for example, from a pressurized pack through a specially shaped seal. 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.

[0305] In some embodiments, the agents described herein can be formulated as inhalants. The inhalation method can deliver the drug directly to the airways. The agents can be formulated as aerosols, microspheres, liposomes, or nanoparticles. The agents can be formulated with a solvent, gas, nitrate, or any combination thereof. The compositions described herein are optionally formulated for delivery as a liquid aerosol or an inhalable dry powder. The liquid aerosol formulation is optionally atomized primarily to a particle size that can be delivered to the terminal bronchioles and respiratory bronchioles. The 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.

[0306] The aerosolized formulations described herein are optionally delivered using an aerosol-forming device, such as a nebulizer, vibrating porous plate, or ultrasonic nebulizer, preferably selected to allow formation of an aerosol particle size with a mass median aerodynamic diameter primarily between 1 and 5 μm. Additionally, the formulation preferably has a balanced osmotic pressure, ionic strength, and chloride concentration, as well as a minimum aerosolizable volume capable of delivering an effective dose of the agent. Additionally, the aerosolized formulation preferably does not cause negative impairment of airway function and does not cause undesirable side effects.

[0307] Aerosolization devices suitable for administering the aerosol formulations described herein include, for example, jet nebulizers, vibrating porous plate types, ultrasonic nebulizers, and electrically powered dry powder inhalers, which are capable of aerosolizing the formulation into an aerosol particle size mainly in the range of 1 - 5 μm. In the present application, "mainly" means that at least 70%, but preferably more than 90%, of all the aerosol particles produced are in the range of 1 - 5 μm. Jet nebulizers operate by air pressure to break a liquid solution into aerosol droplets. Vibrating porous plate nebulizers operate by using acoustic vacuum generated by a rapidly vibrating porous plate to extrude solvent droplets through the porous plate. Ultrasonic nebulizers operate by piezoelectric crystals (shearing 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 and Pari LC jet nebulizers (Pari Respiratory Equipment, Inc., Richmond, Virginia) and Aerosonic™ (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany) and (Omron Healthcare, Inc., Vernon Hills, Illinois) ultrasonic nebulizers.

[0308] In some embodiments, the medicament can be formulated with an oily base or an ointment to form a semi-solid composition having a desired shape. In addition to the medicament, these semi-solid compositions can also contain dissolved and / or suspended bactericides, preservatives, and / or buffer systems. The petrolatum component that can be included can be paraffin wax of any viscosity range, from mineral oil admixed with isobutene, colloidal silica, or stearate to paraffin wax. Absorbent bases can be used with the oily system. Additives can include cholesterol, lanolin (lanolin derivatives), beeswax, fatty alcohols, lanolin alcohols, low HLB (hydrophobic-lipophilic balance) emulsifiers, and various ionic and non-ionic surfactants, used alone or in combination.

[0309] Controlled-release or sustained-release transdermal or topical formulations can be achieved by adding timed-release additives available in the art, such as polymeric structures, matrices. For example, the composition can be administered by using a hot melt extrusion product, such as a bioadhesive hot melt extrusion film. The formulation can comprise a crosslinked polycarboxylic acid polymer formulation. The crosslinking agent can be present in an amount that provides sufficient adhesion to keep the system attached to the surface of the target epithelial or endothelial cells for a sufficient time to allow the desired release of the compound.

[0310] Inserts, transdermal patches, bandages or articles can comprise a mixture or coating of polymers that provide release of the medicament at a constant rate over an extended 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.

[0311] Transdermal devices (inserts, patches, bandages) can also comprise water-insoluble polymers. Rate-controlling polymers can be used for administration to sites where pH changes can be used to achieve release. These rate-controlling polymers can be applied using a continuous coating film during the spraying and drying process with the active compound. In one embodiment, a coating formulation is used to coat a pellet comprising the active ingredient, which pellet is compressed to form a solid, biodegradable insert.

[0312] Polymeric formulations can also be used to provide controlled or sustained release. Bioadhesive polymers described in the art can be used. For example, sustained-release gels and compounds can be incorporated into a polymeric matrix, such as a hydrophobic polymeric matrix. Examples of polymeric matrices include microparticles. The microparticles can be microspheres, and the core can have a different material from the polymeric shell. Alternatively, the polymer can be cast into thin sheets 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, scaffold, 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.

[0313] A kit providing a unit dose of one or more of the medicaments described herein, typically an oral or injectable dose. Such kits may comprise a container containing the unit dose, an informational package insert describing the use of the drug in treating a disease and the attendant benefits, and optionally an appliance or device for delivering the composition.

[0314] Methods of treatment

[0315] In one aspect, the present disclosure provides a method of treating a subject having a tumor. In some cases, treatment of a subject having a tumor comprises administering to a subject in need thereof a compound or salt of any one of Formula (I), Formula (I-A), Formula (I-B), Formula (II), Formula (III), Formula (III-A), Formula (IV-A), Formula (IV-B), Formula (IV-C), or Formula (IV-D), or a pharmaceutical composition of any one of them. 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.

[0316] 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 tree, particularly hepatocellular carcinoma; bowel cancers, 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); central nervous system tumors, particularly brain cancer; multiple myeloma (MM), lymphomas, such as Hodgkin's lymphoma, lymphoplasmacytic lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue lymphoma, mantle cell lymphoma, B-cell large cell lymphoma, Burkitt's 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 associated 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 methods can be repeated as needed. The cancer can be kidney cancer, lung cancer, gastric cancer, or ovarian cancer.

[0317] In some embodiments, treatment of a tumor-bearing subject, relative to an untreated subject, can inhibit tumor growth by at least about 20%, more preferably at least about 40%, even more preferably at least about 60%, and still more preferably at least about 80%. A therapeutically effective amount of a therapeutic compound can reduce tumor size or otherwise improve the symptoms of a subject, which is typically a human but can be another mammal.

[0318] In some embodiments, the compounds described herein can be administered in combination with other therapeutic agents, including antibodies, alkylating agents, angiogenesis inhibitors, antimetabolites, DNA cleaving 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, auristatin, bendamustine, bevacizumab, bicalutamide, bleomycin, bortezomib, busulfan, callistatin A, camptothecin, capecitabine, carboplatin, carmustine, cetuximab, cisplatin, cladribine, cytarabine, cryptophycin, dacarbazine, dasatinib, daunorubicin, docetaxel, doxorubicin, duocarmycin, dynemycin A, epothilone, etoposide, floxuridine, fludarabine, 5-fluorouracil, gefitinib, gemcitabine, ipilimumab, hydroxyurea, imatinib, infliximab, interferon, interleukin, lapachone, lenalidomide, irinotecan, maytansine, mechlorethamine, melphalan, 6-mercaptopurine, methotrexate, mitomycin C, nilotinib, oxaliplatin, paclitaxel, procarbazine, suberoylanilide hydroxamic acid (SAHA), 6-thioguanidine, thiotepa, teniposide, topotecan, trastuzumab, trichostatin A, vinblastine, vincristine, and vindesine.

[0319] The compounds described herein can be used to prepare medicaments for preventing or treating diseases or conditions. In addition, a method for treating any disease or condition described herein in a subject in need of such treatment comprises administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising at least one compound described herein or a pharmaceutically acceptable salt, pharmaceutically acceptable prodrug or pharmaceutically acceptable solvate thereof.

[0320] Compositions containing the compounds described herein can be used 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 inhibit 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 and response of the patient to the drug, and the judgment of the attending physician.

[0321] In prophylactic applications, a composition containing the compounds described herein is administered to a patient who is susceptible to a particular disease, disorder or condition or otherwise at risk of a particular disease, disorder or condition. Such amount is defined as a "prophylactically effective amount or dose". In such use, the exact amount also depends on the health status, weight, etc. of the patient. When used in a patient, the effective amount for this use will depend on the severity and course of the disease, disorder or condition, previous treatment, the health status and response of the patient to the drug, and the judgment of the attending physician.

[0322] In cases where the condition of the patient does not improve, administration of the compound can be made chronically (i.e., for an extended period of time, including throughout the entire duration of the patient's life), at the discretion of the physician, to improve or otherwise control or limit the symptoms of the patient's disease or condition.

[0323] Once improvement of the patient's condition has occurred, a maintenance dose is administered if needed. Subsequently, the dose and / or frequency of administration can be reduced, depending on the symptoms, to a level that maintains the improved disease, disorder or condition. However, the patient may require intermittent treatment on a long-term basis upon any recurrence of symptoms.

[0324] The amount of a given medicament corresponding to that quantity will vary depending on factors such as the specific compound, the disease or condition and its severity, and the identity (e.g., weight) of the subject or host to be treated, but can still be determined in a manner recognized in the art according to the specific circumstances surrounding the situation, which include, for example, the specific medicament administered, the route of administration, the condition being treated, and the subject or host being treated. However, generally speaking, the dosage for adult treatment will typically be in the range of about 0.02 - about 5000 mg per day (in some embodiments about 1 - about 1500 mg per day). The desired dosage can conveniently be presented as a single dose or as separate doses administered simultaneously (or within a short period of time) or at appropriate intervals (e.g., as two, three, four or more sub-doses per day).

[0325] The pharmaceutical compositions described herein can be unit dosage forms suitable for the precise administration of a single dose. In a unit dosage form, the formulation is divided into unit doses containing a suitable amount of one or more compounds. A unit dose can be a packaged form containing a discrete amount of the formulation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. An aqueous suspension composition can be packaged in a single-dose non-refillable container. Alternatively, a multi-dose refillable container can be used, in which case a preservative is typically included in the composition. By way of example only, a formulation for parenteral injection can be presented in unit dosage form (including but not limited to ampoules, or in multi-dose containers with added preservatives).

[0326] The toxicity and therapeutic efficacy of such treatment 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 toxicity and therapeutic effect is the therapeutic index, and it can be expressed as the ratio of LD 50 to ED 50 . Compounds exhibiting a high therapeutic index are preferred. Data obtained from cell culture assays and animal studies can be used to formulate a dosage range for use in humans. The dosage of such compounds is preferably within the circulating concentration range that includes the ED 50 with minimal toxicity. The dosage can vary within this range depending on the dosage form employed and the route of administration used.

[0327] 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 or a pharmaceutically acceptable salt thereof according to any one of the embodiments of the present invention. The disease, condition or disorder can be selected from the descriptions elsewhere herein.

[0328] Preparation of the compound

[0329] 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. For example, the compounds of the present disclosure can be synthesized using the methods described herein and synthetic methods known in the field of synthetic organic chemistry, or variants thereof understood by those skilled in the art. The compounds of the present disclosure can be prepared as described in the schemes and examples described elsewhere herein.

[0330] The following examples further illustrate the invention, but should not of course be construed as limiting its scope in any way.

[0331] Examples

[0332] The following synthetic schemes are provided for illustrative purposes only. The following examples illustrate various methods for preparing the compounds described herein. It should be understood that those skilled in the art can 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.

[0333] Example 1: Synthesis of Compound 7

[0334]

[0335] Compound 3: A mixture of Compound 1 (211 mg, 1.52 mmol), p-toluenesulfonic acid (143 mg, 0.76 mmol), and Compound 2 (400 mg, 1.52 mmol) was refluxed in 20 mL of toluene for 24 h. The solvent was removed in vacuo, and the remaining residue was purified by RP-HPLC to give Compound 3 (280 mg) as a brown solid.

[0336] Compound 4: To a solution of Compound 3 (280 mg, 0.76 mmol) in methanol (8 mL) was added water (6 mL), concentrated sulfuric acid (3.4 mL), hydrogen peroxide (30%, 0.66 mL), and ferrous sulfate (278, 1.0 mmol). The mixture was stirred at room temperature for 1 day. The mixture was then directly purified by RP-HPLC to give Compound 4 (277 mg) as a brown solid.

[0337] Compound 5: A solution of Compound 4 (277 mg, 0.70 μmol) in acetic acid (40 mL) was heated to reflux for 3 h. The solvent was evaporated in vacuo, and the resulting residue was purified by RP-HPLC to give Compound 5 (155 mg) as a pale yellow solid.

[0338] Compound 6: To a suspension of Yb(OTf)3 (16 mg, 0.03 mmol) in 5 mL of anhydrous DCM containing molecular sieves was added a solution of Compound 5 (100 mg, 0.25 mmol) in 20 mL of DCM, followed by addition of a solution of hydroxylamine (0.25 mmol) in 5 mL of DCM. The resulting mixture was stirred at room temperature for 2 h. After filtering off the molecular sieves, the solvent was evaporated in vacuo, and the resulting residue was dissolved in 10 mL of methanol. Then 20 mg of 10% Pd / C was added, and the resulting mixture was stirred under a hydrogen atmosphere for 1 h. The catalyst was filtered off, and the solvent was evaporated in vacuo. The resulting residue was purified by RP-HPLC to give Compound 6 as a pale yellow solid (68 mg).

[0339] Compound 7: To a solution of Compound 6 (TFA salt, 20 mg, 39 μmol) in anhydrous DMF (2 mL) was added N-Fmoc-glycine (12 mg, 39 μmol), PyAOP (21 mg, 39 μmol), and DIEA (28 μL, 156 μmol). The mixture was stirred at room temperature for 20 min. Then piperidine (0.2 mL) was added, and stirring was continued for an additional 10 min. Then the mixture was directly purified by RP-HPLC to give Compound 7 as a brown solid (17 mg). MS: 453.3 [M+H] + 。

[0340] Example 2: Synthesis of Compound 8

[0341]

[0342] To a solution of Compound 6 (TFA salt, 20 mg, 39 μmol) in anhydrous DMF (2 mL) was added glycolic acid (3 mg, 39 μmol), PyAOP (21 mg, 39 μmol), and DIEA (28 μL, 156 μmol). The mixture was stirred at room temperature for 20 min. Then the mixture was directly purified by RP-HPLC to give Compound 8 as a brown solid (21 mg). MS: 454.0 [M+H] + 。

[0343] Example 3: Synthesis of Compound 13

[0344]

[0345] Compound 10: To a solution of Compound 9 (280 mg, 0.76 mmol) in methanol (8 mL) was added water (6 mL), concentrated sulfuric acid (3.4 mL), hydrogen peroxide (30%, 0.66 mL), and ferrous sulfate (278, 1.0 mmol). The mixture was stirred at room temperature for 1 day. The mixture was then purified directly by RP-HPLC to give Compound 10 as a brown solid (257 mg).

[0346] Compound 11: A solution of Compound 10 (257 mg, 0.66 μmol) in acetic acid (40 mL) was heated to reflux for 3 h. The solvent was evaporated in vacuo, and the resulting residue was purified by RP-HPLC to give Compound 11 as a pale yellow solid (140 mg).

[0347] Compound 12: To a suspension of Yb(OTf)3 (22 mg, 0.04 mmol) in 5 mL of anhydrous DCM containing molecular sieve was added a solution of Compound 11 (140 mg, 0.35 mmol) in 20 mL of DCM, followed by addition of a solution of hydroxylamine (0.35 mmol) in 5 mL of DCM. The resulting mixture was stirred at room temperature for 2 h. After filtering off the molecular sieve, the solvent was evaporated in vacuo, and the resulting residue was dissolved in 10 mL of methanol. Then 20 mg of 10% Pd / C was added, and the resulting mixture was stirred under a hydrogen atmosphere for 1 h. The catalyst was filtered off, and the solvent was evaporated in vacuo. The resulting residue was purified by RP-HPLC to give Compound 12 as a pale yellow solid (72 mg).

[0348] Compound 13: To a solution of Compound 12 (TFA salt, 20 mg, 39 μmol) in anhydrous DMF (2 mL) was added N-Fmoc-glycine (12 mg, 39 μmol), PyAOP (21 mg, 39 μmol), and DIEA (28 μL, 156 μmol). The mixture was stirred at room temperature for 20 min. Then piperidine (0.2 mL) was added, and stirring was continued for an additional 10 min. The mixture was then purified directly by RP-HPLC to give Compound 13 as a brown solid (20 mg). MS: 451.1 [M+H] + 。

[0349] Example 4: Synthesis of Compound 14

[0350]

[0351] To a solution of compound 12 (TFA salt, 20 mg, 39 μmol) in anhydrous DMF (2 mL) was added glycolic acid (3 mg, 39 μmol), PyAOP (21 mg, 39 μmol) and DIEA (28 μL, 156 μmol). The mixture was stirred at room temperature for 20 min. The mixture was then purified directly by RP-HPLC to give compound 14 as a brown solid (14 mg). MS: 452.4 [M+H] + 。

[0352] Example 5: Cytotoxicity measurement

[0353] The ability of the compound to inhibit cell growth was measured using an in vitro cytotoxicity assay.

[0354] Cells in logarithmic growth phase were cultured and seeded into 96-well plates. Each cell line was plated at slightly different concentrations, but in the range of 5×10 3 to 50×10 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 reagent (Promega Inc.) at room temperature for 15 minutes and viability was determined by a photometer. The EC50 values for the breast cancer cell line SKBR3 are shown in Table 1.

[0355] Table 1.

[0356]

Claims

1. A compound of formula (I): or a pharmaceutically acceptable salt thereof, wherein: L 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 Residues selected from those containing 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 1 selected from -O- and -NR 7 -; R 2 selected from optionally substituted 5- to 6-membered heterocycles, wherein the 5- to 6-membered heterocycles are 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; 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 groups; Each R 5 is independently selected from sugar; Each R 6 is 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; R 7 selected from hydrogen and C 1-6 alkyl; R 8 selected from hydrogen and hydroxyl; R 9 selected from hydrogen and halogen, wherein R 8 or R 9 at least one of which is hydrogen; m is selected from 0 and 1; n 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.

2. The compound or salt according to claim 1, wherein formula (I) is represented as or a pharmaceutically acceptable salt thereof.

3. The compound or salt according to claim 1, wherein formula (I) is represented as or a pharmaceutically acceptable salt thereof.

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

6. The compound or salt according to claim 4, wherein L is L 2 -L 3 -L 4 -L 7 -R 2 .

7. The compound or salt according to claim 4, wherein L is L 2 -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 .

8. The compound or salt according to claim 4, wherein L is L 2 -L 2B -L 3 -L 4 -R 2 .

9. The compound or salt according to any one of claims 1 to 8, wherein L 2 is a C1 alkylene group.

10. The compound or salt according to claim 1 or 3, wherein L is L 2B -L 3 -L 4 -R 2 and t is 0.

11. The compound or salt according to claim 1 or 3, wherein L is L 2C -L 3 -L 4 -R 2 .

12. The compound or salt according to claim 1 or 3, wherein L is L 2C -L 3 -L 4 -L 7 -R 2 .

13. The compound or salt according to claim 1 or 3, wherein L is L 2C -L 3 -L 4 -L 5 -L 6 -L 7 -R 2 .

14. A compound or salt according to any one of claims 1, 3 or 10 to 13, wherein L 2C is C(O)O-CH2-phenyl.

15. The compound or salt according to any one of claims 1 to 14, wherein L 3 is selected from residues comprising 1 to 5 amino acids.

16. The compound or salt according to any one of claims 1 to 15, wherein the amino acid is a natural amino acid.

17. The compound or salt according to any one of claims 1 to 16, wherein the amino acid is selected from α-amino acids and β-amino acids.

18. The compound or salt according to claim 17, 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.

19. The compound or salt according to any one of claims 1 to 18, wherein L 3 said amino acid of is selected from glycine and phenylalanine.

20. The compound or salt according to any one of claims 1 to 19, wherein L 3 the residue of which comprises 4 amino acids.

21. A compound or salt according to any one of claims 1 to 9 or 11 - 20, wherein the residue of L 3 comprises at least two different amino acids.

22. The compound or salt according to any one of claims 1 to 9 or 11 to 20, wherein L 3 is 23. The compound or salt according to claim 10, wherein L 3 said amino acid of which is selected from sarcosine.

24. The compound or salt according to claim 23, wherein L 3 the residue of contains 1 amino acid.

25. The compound or salt according to claim 24, wherein L 3 is 26. The compound or salt according to claim 1 or 10, wherein L 2B is selected from (NR 4 ) t C(O)O-C1-alkylene-phenyl, wherein the phenyl is substituted with one R 5 ; t is 1.

27. The compound or salt according to claim 26, wherein R 5 is 28. The compound or salt according to any one of claims 1 to 27, wherein L 4 is selected from -C(O)-(CH2)2- and -C(O)-(CH2)5-.

29. The compound or salt according to claim 1 or 7, wherein L 5 is -(O-CH2-CH2-)4-NH-.

30. The compound or salt according to claim 1 or 7, wherein L 6 is selected from -C(O)-(CH2)2- and -C(O)-(CH2)5-.

31. The compound or salt according to claim 24, wherein L 6 is -C(O)-(CH2)2-.

32. The compound or salt according to any one of claims 1, 6 or 7, wherein L 7 is phenyl.

33. The compound or salt according to any one of claims 1 to 32, wherein R 2 is selected from optionally substituted 5-membered heterocycles.

34. The compound or salt according to claim 33, wherein R 2 is selected from 5-membered heterocycles substituted with at least two oxo groups.

35. The compound or salt according to claim 34, wherein R 2 is 36. The compound or salt according to claim 1 or 2, wherein L is selected from 37. The compound or salt according to claim 1 or 3, 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 38. The compound or salt according to any one of claims 1 to 37, which further comprises a ligand.

39. The compound or salt according to any one of claims 1 to 38, wherein the compound or salt is further modified by a ligand.

40. The compound or salt according to any one of claims 1 to 39, wherein the compound or salt is covalently attached to a ligand.

41. The compound or salt according to any one of claims 1 to 40, wherein the compound or salt reacts with a ligand to form a covalent bond.

42. The compound or salt according to any one of claims 38 to 41, wherein formula (I) is represented as or a pharmaceutically acceptable salt thereof, wherein; Lg is the ligand.

43. A compound of formula (III): or a pharmaceutically acceptable salt thereof, wherein: Lg is a ligand; L 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-C 1-6 alkylene-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 by halogen, -OH, -CN, -NO2, -NH2, oxo, -C 1-10 haloalkyl, -O-C 1-10 alkyl, C 2-10 alkenyl, C 2-10 alkynyl substitution; L 3 Residues selected from those containing 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 1 selected from -O- and -NR 7 -; 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; R 3 Selected from hydrogen and C 1-6 alkyl group; R 4 selected from hydrogen and optionally C alkyl substituted by one or more SO2C 1-6 alkyl 1-6 alkyl; Each R 5 is independently selected from sugar; R 7 selected from hydrogen and C 1-6 alkyl; R 8 selected from hydrogen and hydroxyl; R 9 selected from hydrogen and halogens, wherein R 8 or R 9 at least one of which is hydrogen; m is selected from 0 and 1; n 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.

44. The compound or salt according to claim 43, wherein formula (III) is represented as or a pharmaceutically acceptable salt thereof.

45. The compound or salt according to any one of claims 38 to 44, wherein the ligand is selected from an antibody or an antigen-binding fragment thereof.

46. The compound or salt according to claim 45, wherein the ligand is selected from a chimeric antibody, a humanized antibody, and a fully human antibody.

47. A compound of formula (IV-A): or a pharmaceutically acceptable salt thereof.

48. A compound of formula (IV-B): or a pharmaceutically acceptable salt thereof.

49. A compound of formula (IV-C): or a pharmaceutically acceptable salt thereof.

50. A compound of formula (IV-D): or a pharmaceutically acceptable salt thereof.

51. A pharmaceutical composition comprising the compound or salt according to any one of claims 1 to 50 and a pharmaceutically acceptable excipient.

52. Use of the compound or salt according to any one of claims 1 to 50, or the pharmaceutical composition according to claim 51, in the treatment of tumors.

53. Use of the compound or salt according to any one of claims 1 to 50, or the pharmaceutical composition according to claim 51, in the treatment of cancer.

54. A method of treating a subject suffering from a tumor, comprising administering to the subject in need thereof the compound or salt according to any one of claims 1 to 50, or the pharmaceutical composition according to claim 51.

55. A method of treating a subject suffering from cancer, comprising administering to the subject in need thereof the compound or salt according to any one of claims 1 to 50, or the pharmaceutical composition according to claim 51.

56. The method according to claim 55, 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.

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