Cytotoxic compounds

Abstract

The present disclosure provides, inter alia, glycosylated compounds and conjugates thereof useful in the treatment of cancer.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 387,426, filed December 14, 2022, which is incorporated herein by reference in its entirety.

[0002] The present disclosure relates to DNA alkylation units comprising alkylation linkage units. In particular, it relates to compounds comprising G alkylation units (e.g., PBD, PDD, or any other suitable units) or A alkylation units (e.g., CXI units), as well as pharmaceutically acceptable salts thereof, which are useful as pharmaceuticals, particularly as antiproliferative agents. [Background technology]

[0003] Pyrrolobenzodiazepines (PBDs) are a group of compounds, some of which have been shown to be sequence-selective DNA minor groove binders. PBDs were originally discovered in Streptomyces species. [1] PBDs are tricyclic in nature, consisting of fused 6-, 7-, and 5-membered rings, and can be identified as anthranilates (ring A), diazepines (ring B), and pyrrolidines (ring C). [1c] PBDs are characterized by an electrophilic N₁₀=C₁₁ imine group (shown below) or its hydrated equivalent, carbinolamine [NH—CH(OH)], or carbinolamine alkyl ether ([NH—CH(OR, where R=alkyl)]), which can form a covalent bond to the C₂-amino group of guanine in DNA to form a DNA adduct. [2] This natural product interacts with excellent fit (i.e., good "isohelicity") within the minor groove of the DNA helix due to the right-handed longitudinal twist induced by the stereogenic C11a-position with (S) configuration. [3] . [ka] Scheme 1: Interconversion forms of PBD

[0004] DNA adducts bind transcription factors [4][5] and endonucleases [6] and RNA polymerase [7] PBD monomers (e.g., anthramycin) span three base pairs and have a thermodynamic preference for the sequence 5'-Pu-G-Pu-3' (where Pu = purine and G is the reactive guanine).

[11] and a kinetic preference for the sequence 5'-Py-G-Py-3' (where Py is pyridine) as determined by footprinting. [3] , NMR [8] , molecular modeling [9] , and X-ray crystallography

[10] It has been shown by

[0005] PBDs are thought to interact with DNA primarily by locating low-energy binding sequences (i.e., 5'-Pu-G-Pu-3' triplets) through van der Waals, hydrogen bonding, and electrostatic interactions. [4] Then, once in place, nucleophilic attack by the exocyclic C2-amino group of the central guanine occurs, forming a covalent adduct. [4] (Figure 2). Once bound, the PBD remains anchored in the minor groove of the DNA, causing a slight distortion of the DNA helix, thereby evading DNA repair.

[10] The ability of PBDs to form adducts in the minor groove and the ability of PBD dimers to cross-link DNA allows them to interfere with DNA processing and thus their potential use as antiproliferative agents.

[0006] WO 2017 / 032983A, WO 2013 / 164592A, WO 2017 / 223275A, WO 2021 / 137646A, WO 2019 / 126691A, WO 2019 / 104289A, U.S. Pat. No. 10,526,294, and U.S. Pat. No. 10,143,695 disclose PBD (6-7-5) and pyridinobenzodiazepine (PDD, 6-7-6) monomers linked to heterocyclic chains via their A rings, all of which have been shown to act as cytotoxic agents in vitro and as antitumor agents in animal tumor models in vivo. Additionally, the C8'-linked PBD dimer SJG-136

[12] has completed Phase I clinical trials for leukemia and ovarian cancer.

[13] A number of PBD dimer-based antibody-drug conjugates (ADCs) have been designed, some of which are in various stages of clinical trials. [ka] Scheme 2: Chemical structure of SJG-136

[0007] In addition, the indolinobenzodiazepine ADCs IMGN779 and IMGN632

[14] are progressing into Phase II and Phase III trials, respectively. [ka] Scheme 3: Chemical structure of IMGN-779

[0008] U.S. Patent Application Publication No. 2019 / 151465A (LegoChem) discloses antibody-drug conjugates (ADCs) in which multiple active agents are conjugated to an antibody via at least one branched linker. International Publication No. 2020 / 222573A (LegoChem) discloses a tris-type linker. International Publication No. 2018 / 234636A (Glykos) discloses hydrophilic linkers and conjugates.

[0009] Sibiromycin, a naturally occurring PBD monomer

[15] is one of the most potent natural PBDs reported (submicromolar cytotoxicity) and contains a sibirosamine sugar at the C7 position of the molecule. Its potent cytotoxicity is thought to be related to its DNA binding profile and potential ability to inhibit transcription factor binding.

[16] .

[0010] The carbohydrate moiety has previously been incorporated into PBD-based monomers developed by Lown and coworkers.

[17] The addition of sugar moieties to specific positions on the polyamide chain was observed to enhance cytotoxicity in some cell lines.

[0011] Glucuronides and glucose are known to be cleaved by glycosidases in vivo and have been incorporated as part of linker constructs within various families of both amine- and phenol-containing linker-payloads.

[18] Incorporation of glucose in this context enhances the PK properties of the resulting ADC, and the payload is released from the linker-payload construct through cleavage of the glucuronide moiety. Glucose moieties have also been incorporated into auristatin (e.g., monomethyl auristatin E, "MMAE")-based prodrugs.

[19] , which has enhanced efficacy, tolerability, and solubility when compared to the parent, unsubstituted MMAE molecule.

[0012] Other sequence-selective DNA minor groove binders are known, including duocarmycins or CXIs (eg, cyclopropapyrroloindole (CPI), cyclopropabenzindole (CBI), or cyclopropathienoindole (CTI) moieties). 1

[0013] Many CXI analogs have since been developed, including bis-alkylated dimers of two CXI units (AA interchain crosslinkers), such as bizelesin. [ka] Scheme 4: Chemical structure of the CPI dimer Bizelesin

[0014] Bizelesin is an example of a seco-CPI (open-ring form, with a chloromethyl functionality) rather than the spiro (closed-ring cyclopropyl) form found in CC-1065. 5

[0015] This seco modification was found to be a prodrug of CPI, and was more stable but equally active as the CPI dimer U-77809 (the corresponding parent drug form). The prodrug (seco) CXI form is known to spirocyclize to the cyclopropane-active spiro form via a Winstein-Baird mechanism (Scheme 5). [ka] Scheme 5: Interconvertible forms of CXI

[0016] WO 2015 / 104373A and WO 2015 / 104386A (Synthon), WO 2017 / 012924A (Nerviano), and WO 2003 / 0022806A (Boger) also disclose CXI moieties.

[0017] The excellent potency of the PBD(6-7-5) and PDD(6-7-6) compounds and CXI compounds makes them attractive candidates for targeted delivery via antibody-drug conjugates (ADCs).

[0018] The present disclosure seeks to address this need and overcome problems associated with the prior art. Summary of the Invention

[0019] In one aspect, the inventors surprisingly discovered that carbohydrate (e.g., sugar) moieties can be successfully incorporated into specific positions of the payload structure of an extended G- or A-alkylating agent, thereby enhancing the hydrophilicity of the payload itself. The specific positions selected allow for the addition of sugar groups to the molecule without adversely interfering with the released drug's ability to kill cancer cells. This is advantageous because substitution at specific positions of the heterocyclic chain on the payload can alter DNA binding and cytotoxicity. As a non-limiting example, Figure 21 shows non-limiting examples of second-generation linker payloads bearing sugar moieties at specific positions on G-alkylated compounds. In the case of the subject substances, the parent molecule after release can effectively bind to DNA and result in potent cytotoxicity. Because the glycosylated substituent can act as a prodrug moiety, which is selectively cleaved at the tumor site, ADCs containing glycosylated payloads can produce potent in vivo efficacy (similar to that of the unsubstituted parent molecule) but with a substantially increased tolerability profile, thereby substantially expanding the therapeutic window. In some embodiments, compounds described herein and / or conjugates thereof containing at least one glycosylation substituent exhibit enhanced or increased hydrophilicity compared to compounds that do not contain at least one glycosylation substituent. In some embodiments, compounds described herein and / or conjugates thereof containing at least one glycosylation substituent exhibit enhanced or increased tolerability compared to compounds that do not contain at least one glycosylation substituent. In combination, these qualities result in highly effective compounds. Thus, the inventors have discovered that incorporating a sugar moiety into the structure of a G-monoalkylating agent or an A-monoalkylating agent provides advantageous properties for efficient conjugation, efficacy, and tolerability.

[0020] Nevertheless, there is a need for improved cytotoxic payloads, payloads that may be particularly useful in ADCs. For example, when the linker-payload N-(4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)propanamido)phenyl)-4-(4-(((S)-2-methoxy-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepin-3-yl)oxy)butanamido)-1-methyl-1H-pyrrole-2-carboxamide (i.e., Control 3) is conjugated to multiple antibodies at DAR4, the MTD of the established single-dose "platform" is 50 mg / kg, with an effective dose of 10 mg / kg, and regression observed at 5 mg / kg. Given the hydrophobicity of this construct, control 3 cannot be used to produce ADCs with higher DARs (which would have a better bystander effect than DAR4).

[0021] Thus, the present disclosure provides, in a first aspect, a compound of formula (I): DQBT (I) or a salt, solvate, or tautomer thereof, wherein: D is a source of alkylated DNA minor groove binding units, Q is a linker, B is a DNA-binding amide-containing chain, T is a terminal group, wherein D, B, Q and / or T comprise at least one carbohydrate substituent.

[0022] Suitably, the carbohydrate substituent is a monovalent saccharide substituent, preferably R S It could be.

[0023] Suitably, D may be a G alkylating agent unit G comprising a group of formula (II): [ka] During the ceremony, The dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4; The wavy line indicates the point of attachment to Q; m is 0 or 1, R1, R3, and R4 are independently selected from H and R 29 and R2 is selected from H, L2-R 28、 R 29 , and -L S -R S or one of R1 and R2, R2 and R3, or R3 and R4, taken together with the carbon atom to which they are attached, is selected from a 6-membered aryl, or one, two, or three independently selected optional R 20 forming a 5- or 6-membered cyclic, heterocyclic or heteroaryl ring optionally substituted by a group, R5 and R6 are: (i) R5 is H, OH, and OC 1-6 alkyl, and R6 is selected from H, SO3H, -L S -R S , nitrogen protecting group, -L2-R 28 and R A or (ii) R5 is oxo or H and R6 is H or C 1-6 alkyl, or (iii) R5 and R6 together form a double bond; R7 and R9 are independently H and R 20 is selected from R8 is H, SR 24 , SCH2Ph, R 20 , L2-R 28 , and -L S -R S is selected from R A is (CH2) j -OH, (CH2) j -CO2R 26 , C(=O)-O-(CH2) k -NR 26 R 27 , (CH2) jNR 26 R 27 , C(=O)-NH-(CH2) j -NR 26 R 27 and C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 is selected from L2 is a bond or a linker moiety having 1 to 200 non-hydrogen atoms selected from C, N, P, O, S or halogen, and may incorporate an ether, oxo, carboxamidyl, uretanyl, branched, cyclic, unsaturated, heterocyclyl, aryl, or heteroaryl moiety; R 28 are azides, alkynes, bisulfones, carbohydrazides, hydrazines, hydroxylamines, iodoacetamides, isothiocyanates, maleimides, phosphines, pyridopyridazines, semihydrazides, succinimidyl esters, sulfodichlorophenol esters, sulfonyl halides, sulfosuccinimidyl esters, 4-sulfotetrafluorophenyl esters, tetrafluorophenyl esters, thiazoles, R A , O-(CH2) k -NR 26 R 26 , NHNH2, or a targeting agent selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; Each R 29 are independently, R 20 , R 21 , =CH2, =CH-(CH2) s -CH3, =CH-(CH2) s -R 21 , =O, (CH2) s -OR 21 , (CH2) s -CO2R 21 , (CH2) s -NR 21 R 24 , O-(CH2) t -NR21 R 24 , NH-C(O)-R 21 , O-(CH2) t -NH-C(O)-R 21 , O-(CH2) t -C(O)-NH-R 21 , (CH2) s -SO2R 21 , O-SO2R 21 , (CH2) s -C(O)R 21 and (CH2) s -C(O)NR 21 R 24 is selected from Each R 20 are independently F, Cl, Br, (CH2) j -OH, C 1-6 Alkyl, OC 1-6 Alkyl, OCH2Ph, (CH2) j -CO2R 26 , O-(CH2) k -NR 26 R 27 , C(=O)-O-(CH2) k -NR 26 R 27 , C(=O)-NR 26 R 27 , (CH2) j -NR 26 R 27 , N.R. 26 NH2, C(=O)-NH-(CH2) j -NR 26 R 27 , C(=O)-NH-C6H4-(CH2) j -R 26 , C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 , -L2-R 28 , S(O)2-(C 1-6 alkyl), O-(CH2) k -O-(C 1-6 alkyl), (CH2) j -S(O)2-NR 26 R 27 , C(=NH)-O-(C 1-6alkyl), (CH2) k -O-(C 1-6 alkyl), CN, NCO, Cy, C(O)-NH-(CH2) j -Cy, C(O)-Cy, NH-C(O)-NR 26 R 27 is selected from [ka] each j and s is independently selected from 0, 1, 2, 3, 4, 5, or 6; each k and t is independently selected from 1, 2, 3, 4, 5, or 6; Each R 21 are independently H, C 1-12 Alkyl, C 5-6 Heterocyclyl, C 5-9 Heteroaryl, C 6-15 Heteroarylalkyl, phenyl, and C 7-12 and aralkyl groups, wherein the heterocyclyl, heteroaryl, heteroarylalkyl, phenyl, and aralkyl groups are selected from one, two, or three independently selected optional R 20 may be substituted with a group, Each R 24 , R 26 , and R 27 are independently H and C 1-12 alkyl, Each Cy independently represents C 5-6 Heterocyclyl or C 5-6 heteroaryl groups, wherein the heterocyclyl or heteroaryl group is selected from one or two R 20 may be substituted with a group, L S is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are P, O, S, NH, C 5-9which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L S teeth, [ka] and R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0024] Suitably, D may be a G alkylating agent unit G comprising a group of formula (II): [ka] During the ceremony, The dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4; The wavy line indicates the point of attachment to Q; m is 0 or 1, R1, R3, and R4 are independently selected from H and R 29 and R2 is selected from H, L2-R 28、 R 29 , and -L S -R S or one of R1 and R2, R2 and R3, or R3 and R4, taken together with the carbon atom to which they are attached, is selected from a 6-membered aryl, or one, two, or three independently selected optional R 20 forming a 5- or 6-membered cyclic, heterocyclic or heteroaryl ring optionally substituted by a group, R5 and R6 are: (i) R5 is H, OH, and OC 1-6 alkyl, and R6 is selected from H, SO3H, -L S -R S , nitrogen protecting group, -L2-R28 and R A or (ii) R5 is oxo or H and R6 is H or C 1-6 alkyl, or (iii) R5 and R6 together form a double bond; R7 and R9 are independently H and R 20 is selected from R8 is H, SR 24 , SCH2Ph, R 20 , L2-R 28 , and -L S -R S is selected from R A is (CH2) j -OH, (CH2) j -CO2R 26 , C(=O)-O-(CH2) k -NR 26 R 27 , (CH2) j NR 26 R 27 , C(=O)-NH-(CH2) j -NR 26 R 27 and C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 is selected from L2 is a bond or a linker moiety having 1 to 200 non-hydrogen atoms selected from C, N, P, O, S or halogen, and may incorporate an ether, oxo, carboxamidyl, uretanyl, branched, cyclic, unsaturated, heterocyclyl, aryl, or heteroaryl moiety; R 28are azides, alkynes, bisulfones, carbohydrazides, hydrazines, hydroxylamines, iodoacetamides, isothiocyanates, maleimides, phosphines, pyridopyridazines, semihydrazides, succinimidyl esters, sulfodichlorophenol esters, sulfonyl halides, sulfosuccinimidyl esters, 4-sulfotetrafluorophenyl esters, tetrafluorophenyl esters, thiazoles, R A , O-(CH2) k -NR 26 R 26 , NHNH2, or a targeting agent selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; Each R 29 are independently, R 20 , R 21 , =CH2, =CH-(CH2) s -CH3, =CH-(CH2) s -R 21 , =O, (CH2) s -OR 21 , (CH2) s -CO2R 21 , (CH2) s -NR 21 R 24 , O-(CH2) t -NR 21 R 24 , NH-C(O)-R 21 , O-(CH2) t -NH-C(O)-R 21 , O-(CH2) t -C(O)-NH-R 21 , (CH2) s -SO2R 21 , O-SO2R 21 , (CH2) s -C(O)R 21 and (CH2) s -C(O)NR 21 R 24 is selected from Each R 20are independently F, Cl, Br, (CH2) j -OH, C 1-6 Alkyl, OC 1-6 Alkyl, OCH2Ph, (CH2) j -CO2R 26 , O-(CH2) k -NR 26 R 27 , C(=O)-O-(CH2) k -NR 26 R 27 , C(=O)-NR 26 R 27 , (CH2) j -NR 26 R 27 , N.R. 26 NH2, C(=O)-NH-(CH2) j -NR 26 R 27 , C(=O)-NH-C6H4-(CH2) j -R 26 , C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 , -L2-R 28 , S(O)2-(C 1-6 alkyl), O-(CH2) k -O-(C 1-6 alkyl), (CH2) j -S(O)2-NR 26 R 27 , C(=NH)-O-(C 1-6 alkyl), (CH2) k -O-(C 1-6 alkyl), CN, NCO, Cy, C(O)-NH-(CH2) j -Cy, C(O)-Cy, NH-C(O)-NR 26 R 27 is selected from [ka] each j and s is independently selected from 0, 1, 2, 3, 4, 5, or 6; each k and t is independently selected from 1, 2, 3, 4, 5, or 6; Each R 21 are independently H, C1-12 Alkyl, C 5-6 Heterocyclyl, C 5-9 Heteroaryl, C 6-15 Heteroarylalkyl, phenyl, and C 7-12 and aralkyl groups, wherein the heterocyclyl, heteroaryl, heteroarylalkyl, phenyl, and aralkyl groups are selected from one, two, or three independently selected optional R 20 may be substituted with a group, Each R 24 , R 26 , and R 27 are independently H and C 1-12 alkyl, Each Cy independently represents C 5-6 Heterocyclyl or C 5-6 heteroaryl groups, wherein the heterocyclyl or heteroaryl group is selected from one or two R 20 may be substituted with a group, L S is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are P, O, S, NH, C 5-9 which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L S teeth, [ka] and L Cis an amino acid, an amino acid derivative, a peptide chain having 2 to 6 amino acids or amino acid derivatives, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-8 -, and these chains contain one or more groups selected from one or more P, O, S and / or NH groups, and / or C 5-9 may be interrupted by heteroarylene and / or phenylene, and each C 5-9 the heteroarylene group and / or each phenylene group may be optionally substituted; R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0025] Suitably, G is selected from groups of formulae G1 to G8 below. [ka] [ka] [ka]

[0026] In some embodiments, D can include an A that is an A-alkylated DNA group of formula (IIIa) or (IIIb): [ka] During the ceremony, Z1 is a leaving group, optionally a halide, triflate, or tosylate; X is CR 17 , N, NR 17 , S or O, and the dashed line to X indicates the optional presence of a double bond depending on the nature of X, R 17 is H, -L S -R S , or R20 and R 20 are independently F, Cl, Br, (CH2) j -OH, C 1-6 Alkyl, OC 1-6 Alkyl, OCH2Ph, (CH2) j -CO2R 26 , O-(CH2) k -NR 26 R 27 , C(=O)-O-(CH2) k -NR 26 R 27 , C(=O)-NR 26 R 27 , (CH2) j -NR 26 R 27 , N.R. 26 NH2, C(=O)-NH-(CH2) j -NR 26 R 27 , C(=O)-NH-C6H4-(CH2) j -R 26 , C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 , -L2-R 28 , S(O)2-(C 1-6 alkyl), O-(CH2) k -O-(C 1-6 alkyl), (CH2) j -S(O)2-NR 26 R 27 , C(=NH)-O-(C 1-6 alkyl), (CH2) k -O-(C 1-6 alkyl), CN, NCO, Cy, C(O)-NH-(CH2) j -Cy, C(O)-Cy, NH-C(O)-NR 26 R 27 is selected from [ka] L2 is a bond or a linker moiety having 1 to 200 non-hydrogen atoms selected from C, N, P, O, S or halogen, and may incorporate an ether, oxo, carboxamidyl, uretanyl, branched, cyclic, unsaturated, heterocyclyl, aryl, or heteroaryl moiety; R 28 are azides, alkynes, bisulfones, carbohydrazides, hydrazines, hydroxylamines, iodoacetamides, isothiocyanates, maleimides, phosphines, pyridopyridazines, semihydrazides, succinimidyl esters, sulfodichlorophenol esters, sulfonyl halides, sulfosuccinimidyl esters, 4-sulfotetrafluorophenyl esters, tetrafluorophenyl esters, thiazoles, R A , O-(CH2) k -NR 26 R 26 , NHNH2, or a targeting agent selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; R A is (CH2) j -OH, (CH2) j -CO2R 26 , C(=O)-O-(CH2) k -NR 26 R 27 , (CH2) j NR 26 R 27 , C(=O)-NH-(CH2) j -NR 26 R 27 and C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 is selected from Each R 26 and R 27 are independently H and C 1-12 alkyl, Each Cy independently represents C 5-6 Heterocyclyl or C5-6 heteroaryl groups, wherein the heterocyclyl or heteroaryl group is selected from one or two R 20 may be substituted with a group, each j is independently selected from 0, 1, 2, 3, 4, 5, or 6; each k is independently selected from 1, 2, 3, 4, 5, or 6; z is 0 or 1, R''' is OH or -L S -R S and L S is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are P, O, S, NH, C 5-9 which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L S teeth, [ka] and R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0027] Suitably, the compound of formula (I) may be a compound of formula IV: [ka] wherein the substituents have the definitions given above.

[0028] Suitably, the linker group Q may comprise X1-L-X2, During the ceremony, X1 is O, S, NR 13 , C.R. 13 R 14 , C.R. 13 R 14 O, C(=O), C(=O)NR 13 , N.R. 13 selected from C(═O), OC(O), and C(O)—O, or absent; L is an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 -, these chains being selected from one or more P, O, S and / or NH groups, and / or C 5-9 may be interrupted by heteroarylene and / or phenylene, and each C 5-9 the heteroarylene group and / or each phenylene group may be optionally substituted; X2 is O, S, NR 15 , C.R. 15 R 16 , C.R. 15 R 16 O, C(=O), C(=O)NR 15 , N.R. 15 selected from C(═O), OC(O), and C(O)—O, or absent; R 13 , R 14、 R 15 , and R 16 are independently H and C 1-6 alkyl.

[0029] Preferably, the DNA-binding amide-containing strand B is (A) q may include During the ceremony, q is selected from 0, 1, 2, 3, 4, 5, and 6; A is selected from the following: [ka] For each A group, one of Y and Y is independently NR 30 , S, and O; the other of Y3 and Y4 is CH; and Y5 is independently selected from CR 30 , N, S, and COH; For each A2 group, one of Y6 and Y7 is independently selected from N and CH, and the other of Y6 and Y7 is CR 30 and Each R 30 are independently H, C 1-6 Alkyl, L2-R 28 , and R S is selected from.

[0030] Suitably, the terminal group T may comprise a group of the formula: [ka] During the ceremony, p is 0 or 1, R T -L2-R 28 , phenyl, and C 5-9 heteroaryl, selected from phenyl and C 5-9 The heteroaryl group optionally includes OH, C 1-6 Alkyl, OC 1-6 Alkyl, -L2-R 28 , (CH2) j -CO2R 11 , O-(CH2) k -NR 11 R 12 , (CH2) j -NR 11 R 12 , C(=O)-NH-(CH2) k -NR 11 R 12 , C(=O)-NH-R 24 , and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12and optionally, but optionally substituted with up to three optional substituents selected from 5-9 heteroaryl is other than indolyl; R 19 is H, C 1-6 Alkyl, L2-R 28 , R S , and (CH2) t -NR 20 R 21 is selected from Y1 and Y2 are independently N or CR 31 and at least one of Y1 and Y2 is CR 31 and Each R 31 are independently H, C 1-6 Alkyl, L2-R 28 , and R S is selected from R 11 , R 12 , and R 24 are independently H, -L2-R 28 , and C 1-6 alkyl.

[0031] Non-limiting examples of compounds of the present disclosure include: [ka] [ka] [ka] [ka] [ka] [ka] [ka] In the formula, R Sis a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0032] Preferably, the compound has at least one L2-R 28 More preferably, D, T, Q and / or B may comprise a L2-R group. 28 It may be substituted with a group.

[0033] L2 is [ka] may be selected from In the formula, X AA is the amino acid sequence, and K2 is -[CH2CH2O] 0-50 -or- [CH2] 0-12 -It is.

[0034] In embodiments, X AA may be L-valyl-L-alanine.

[0035] Preferably, R 28 may be the following maleimide: [ka] It may be linked to a targeting agent.

[0036] Preferably, L2-R 28 teeth, [ka] [ka] may include [ka] may include [ka] or [ka] and may be linked to a targeting agent.

[0037] The compounds according to formula (I) comprise at least one L2-R 28 The targeting agent may comprise a L2-R 28 The compound is linked to the compound via a group.

[0038] In a further aspect, there is provided a compound of formula (I), and salts and solvates thereof, for use as a pharmaceutical.

[0039] In a further aspect, there is provided compounds of formula (I), and salts and solvates thereof, for use in the treatment of a proliferative disease.

[0040] In a further aspect, there is provided compounds of formula (I), and salts and solvates thereof, for use in methods of treatment.

[0041] In a further aspect, there is provided a pharmaceutical composition comprising a compound of formula (I), and salts and solvates, and a pharmaceutically acceptable excipient, carrier, or diluent.

[0042] In a further aspect, there is provided compounds of formula (I), and salts and solvates thereof, for use in the manufacture of a medicament for the treatment of a proliferative disease.

[0043] An ADC (antibody-drug conjugate) can be composed of an antibody, a linker, and a payload. Linker-payload conjugates typically contain hydrophilic groups (in the form of extended PEG) to reduce hydrophobicity and facilitate efficient conjugation of the structure to the antibody. DNA-interacting drugs are typically hydrophobic in nature, posing significant challenges during PK and conjugation processes. The use of sugar groups in the payload structure itself as a masking agent surprisingly reduces hydrophobicity, enhances PK properties, and increases the tolerability of the ADC. This is particularly advantageous because it can result in (a) more efficient conjugation than other classes of payloads, (b) similar cytotoxicity to the unsubstituted form of the payload with significantly improved tolerability and conjugation properties compared to prior art molecules, and (c) an enhanced PK profile compared to the unsubstituted drug.

[0044] Thus, in a further embodiment, the compounds of formula (I), and salts and solvates thereof, may be linked, either directly or indirectly, to a targeting agent to provide the conjugate of interest. In this embodiment, the compound comprises at least one L2-R 28 The targeting agent may comprise a L2-R 28 The compound is linked to the carboxyl group via the carboxyl group.

[0045] The targeting agent may comprise an antibody, an antibody fragment, a hormone or a hormone fragment.

[0046] Therefore, in a further embodiment, the compound of formula (I), as well as its salts and solvates, can be directly or indirectly linked to a targeting agent (e.g., an antibody, an antibody fragment, a hormone, etc.) to provide a conjugate of interest. The conjugate of interest of the present disclosure can contain one or more compounds of formula (I) (or their salts and solvates). Various conjugates of interest are known in the art and can be used with the compound of formula (I) and its salts or solvates. For example, in certain embodiments, the conjugate of interest is an antibody-drug conjugate, in which one or more compounds of formula (I) are directly or indirectly linked to an antibody. Thus, the compound of formula (I), as well as its salts and solvates, can be used as a payload on the conjugate of interest.

[0047] In one aspect, compounds of Formula (I), compounds of Formula (IV), and / or conjugates thereof comprising at least one carbohydrate substituent have enhanced or increased hydrophilicity compared to compounds of Formula (I), compounds of Formula (IV), and / or conjugates thereof that do not comprise at least one carbohydrate substituent. In some embodiments, at least one R S The compounds of formula (I) and / or conjugates thereof may comprise at least one R S In some embodiments, at least one R S The compounds of formula (I), compounds of formula (IV), and / or conjugates thereof, which comprise at least one R S In some embodiments, at least one R S The compounds of formula (I), compounds of formula (IV), and / or conjugates thereof, which comprise at least one R SThe compound of formula (I) has about 1-fold, about 2-fold, about 5-fold, about 10-fold, about 20-fold, about 50-fold, or about 100-fold enhanced or increased hydrophilicity compared to the compound of formula (I), the compound of formula (IV), and / or conjugates thereof that do not contain . In a non-limiting example, the increased or enhanced hydrophilicity allows for efficient conjugation of the compound of formula (I) and / or the compound of formula (IV) to a targeting agent and / or at a higher DAR (e.g., DAR8).

[0048] In some embodiments, compounds of formula (I), compounds of formula (IV), and / or conjugates thereof having a G-alkylated DNA group of formula (II) or one of G1-G8 as the D moiety, which contain at least one sugar chain substituent, have enhanced or increased hydrophilicity compared to compounds of formula (I), compounds of formula (IV), and / or conjugates thereof having a G-alkylated DNA group of formula (II) or one of G1-G8 as the D moiety, which do not contain at least one sugar chain substituent. S The compounds of formula (I) having as the D moiety a G-alkylated DNA group of formula (II) or one of G1 to G8, compounds of formula (IV), and / or conjugates thereof, comprise at least one R S In some embodiments, at least one R S The compounds of formula (I) having as the D moiety a G-alkylated DNA group of formula (II) or one of G1 to G8, compounds of formula (IV), and / or conjugates thereof, comprise at least one R SThe compound of formula (I) has about 1-fold, about 2-fold, about 5-fold, about 10-fold, about 20-fold, about 50-fold, or about 100-fold enhanced or increased hydrophilicity compared to a compound of formula (I), a compound of formula (IV), and / or a conjugate thereof having a G-alkylated DNA group of formula (II) or one of G1-G8 as the D moiety that does not contain: In a non-limiting example, the increased or enhanced hydrophilicity allows for efficient conjugation of a compound of formula (I) having a G-alkylated DNA group of formula (II) or one of G1-G8 as the D moiety and / or a compound of formula (IV) to a targeting agent and / or at a higher DAR (e.g., DAR8).

[0049] In some embodiments, compounds of Formula (I) and / or conjugates thereof having an A-alkylated DNA group of Formula (IIIa) or (IIIb) as the D moiety, which comprises at least one sugar chain substituent, have enhanced or increased hydrophilicity compared to compounds of Formula (I) and / or conjugates thereof having an A-alkylated DNA group of Formula (IIIa) or (IIIb) as the D moiety that does not comprise at least one sugar chain substituent. S Compounds of formula (I) and / or conjugates thereof having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety, including at least one R S In some embodiments, at least one R S Compounds of formula (I) and / or conjugates thereof having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety, including at least one R SThe D moiety of the compound of formula (I) has about 1-fold, about 2-fold, about 5-fold, about 10-fold, about 20-fold, about 50-fold, or about 100-fold enhanced or increased hydrophilicity compared to a compound of formula (I) and / or a conjugate thereof having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety that does not contain: In a non-limiting example, the increased or enhanced hydrophilicity allows for efficient conjugation of a compound of formula (I) having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety to a targeting agent and / or at a higher DAR (e.g., DAR8).

[0050] In one aspect, compounds of formula (I), compounds of formula (IV), and / or conjugates thereof comprising at least one carbohydrate substituent have enhanced or increased tolerability compared to compounds of formula (I), compounds of formula (IV), and / or conjugates thereof not comprising at least one carbohydrate substituent. In some embodiments, at least one R S The compounds of formula (I), compounds of formula (IV), and / or conjugates thereof contain at least one R S In some embodiments, the compound of formula (I), the compound of formula (IV), and / or a conjugate thereof, which does not comprise at least one R S The compounds of formula (I) and / or conjugates thereof may comprise at least one R S In some embodiments, the compound of formula (I) and / or its conjugate has enhanced or increased tolerability compared to a compound of formula (I) and / or a conjugate thereof that does not comprise at least one R S The compounds of formula (I), compounds of formula (IV), and / or conjugates thereof contain at least one R S The tolerability is enhanced or increased by 2 to 10 times or more compared to the compound of formula (I), the compound of formula (IV), and / or a conjugate thereof not containing

[0051] In some embodiments, compounds of formula (I), compounds of formula (IV), and / or conjugates thereof having a G-alkylated DNA group of formula (II) or one of G1-G8 as the D moiety, which contain at least one sugar chain substituent, have enhanced or increased tolerability compared to compounds of formula (I), compounds of formula (IV), and / or conjugates thereof having a G-alkylated DNA group of formula (II) or one of G1-G8 as the D moiety, which do not contain at least one sugar chain substituent. S The compounds of formula (I) having as the D moiety a G-alkylated DNA group of formula (II) or one of G1 to G8, compounds of formula (IV), and / or conjugates thereof, comprise at least one R S In some embodiments, the compounds of formula (I), (IV), and / or conjugates thereof, have enhanced or increased tolerability compared to compounds of formula (I) having a G-alkylated DNA group of formula (II) or one of G1-G8 as the D moiety, which does not contain at least one R S The compounds of formula (I) having as the D moiety a G-alkylated DNA group of formula (II) or one of G1 to G8, compounds of formula (IV), and / or conjugates thereof, comprise at least one R S The tolerability is increased by 2-fold to 10-fold or more compared to a compound of formula (I), a compound of formula (IV), and / or a conjugate thereof having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, which does not contain

[0052] In some embodiments, compounds of formula (I) and / or conjugates thereof having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety, which comprises at least one sugar chain substituent, have enhanced or increased tolerability compared to compounds of formula (I) and / or conjugates thereof having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety that does not comprise at least one sugar chain substituent. SCompounds of formula (I) and / or conjugates thereof having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety, including at least one R S In some embodiments, at least one R S Compounds of formula (I) and / or conjugates thereof having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety, including at least one R S The compounds of formula (I) and / or their conjugates having an A-alkylated DNA group of formula (IIIa) or (IIIb) as the D moiety, which do not contain

[0053] In some embodiments, R6 and / or R8 are S The compounds of formula (I) having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, the compounds of formula (IV), and / or conjugates thereof may contain R at the R6 and / or R8 positions. S The hydrophilicity is enhanced or increased compared to a compound of formula (I), a compound of formula (IV), and / or a conjugate thereof having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, which does not contain

[0054] In some embodiments, R S The compounds of formula (I) having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, the compounds of formula (IV), and / or conjugates thereof may contain R S The tolerability is increased by 2-fold to 10-fold or more compared to a compound of formula (I), a compound of formula (IV), and / or a conjugate thereof having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, which does not contain

[0055] In some embodiments, RS The compounds of formula (I) having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, the compounds of formula (IV), and / or conjugates thereof, may contain R S have the same or substantially the same tolerability as a compound of formula (I), a compound of formula (IV), and / or a conjugate thereof having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, which does not contain [Brief explanation of the drawings]

[0056] Embodiments of the present disclosure will now be further described with reference to the accompanying drawings.

[0057] [Figure 1] Figure 1 shows the results of the β-glucosidase assay at 5 minutes. Aliquots of 100 μL were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 2] Figure 2 shows the results of the β-glucosidase assay at 30 minutes. 100 μL aliquots were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 3] Figure 3 shows the results of the β-glucosidase assay at 1 hour. 100 μL aliquots were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 4] Figure 4 shows the results of the β-glucosidase assay at 2 hours. 100 μL aliquots were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 5] Figure 5 shows the results of the β-glucosidase assay at 18 hours. 100 μL aliquots were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 6] Figure 6 shows the results of the β-glucosidase assay at 36 hours. 100 μL aliquots were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 7] Figure 7 shows the results of the β-glucosidase assay at 60 hours. 100 μL aliquots were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 8] Figure 8 shows the results of the β-glucosidase assay at 84 hours. 100 μL aliquots were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 9] Figure 9 shows the results of the β-glucosidase assay at 132 hours. 100 μL aliquots were taken at 5 minutes, 30 minutes, 1 hour, 2 hours, 18 hours, 36 hours, 60 hours, 84 hours, and 132 hours and analyzed by LC-MS. [Figure 10A] Figure 10A shows the results of the β-galactosidase assay. 100 μL aliquots were taken at 0, 5, 30, 90, 4.5, 7.5, and 20 hours and analyzed by LC-MS. [Figure 10B] Figure 10B shows the results of the β-galactosidase assay: 100 μL aliquots were taken at 0, 5, 30, 90, 4.5, 7.5, and 20 hours and analyzed by LC-MS. [Figure 10C] Figure 10C shows the results of the β-galactosidase assay: 100 μL aliquots were taken at 0, 5, 30, 90, 4.5, 7.5, and 20 hours and analyzed by LC-MS. [Figure 10D] Figure 10D shows the results of the β-galactosidase assay. 100 μL aliquots were taken at 0, 5, 30, 90, 4.5, 7.5, and 20 hours and analyzed by LC-MS. [Figure 10E]Figure 10E shows the results of the β-galactosidase assay. 100 μL aliquots were taken at 0, 5, 30, 90, 4.5, 7.5, and 20 hours and analyzed by LC-MS. [Figure 10F] Figure 10F shows the results of the β-galactosidase assay. 100 μL aliquots were taken at 0, 5, 30, 90, 4.5, 7.5, and 20 hours and analyzed by LC-MS. [Figure 11A] Figure 11A shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(((S)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepin-2-yl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (88). 100 μL aliquots were taken at 0, 5, 30, and 90 minutes and analyzed by LC-MS. [Figure 11B] Figure 11B shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(((S)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepin-2-yl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (88). 100 μL aliquots were taken at 0, 5, 30, and 90 minutes and analyzed by LC-MS. [Figure 11C]Figure 11C shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(((S)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepin-2-yl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (88). 100 μL aliquots were taken at 0, 5, 30, and 90 minutes and analyzed by LC-MS. [Figure 11D] Figure 11D shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(((S)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-12-oxo-6a,7,8,9,10,12-hexahydrobenzo[e]pyrido[1,2-a][1,4]diazepin-2-yl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (88). 100 μL aliquots were taken at 0, 5, 30, and 90 minutes and analyzed by LC-MS. [Figure 12A] Figure 12A shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (138). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 12B]Figure 12B shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (138). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 12C] Figure 12C shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (138). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 12D]Figure 12D shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (138). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 12E] Figure 12E shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (138). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 min and analyzed by LC-MS. [Figure 13A]FIG. 13A shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (146). Aliquots of 100 μL were taken at 0, 5, 30, 90, and 150 min and analyzed by LC-MS. [Figure 13B] Figure 13B shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (146). Aliquots of 100 μL were taken at 0, 5, 30, 90, and 150 min and analyzed by LC-MS. [Figure 13C]Figure 13C shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (146). Aliquots of 100 μL were taken at 0, 5, 30, 90, and 150 min and analyzed by LC-MS. [Figure 13D] Figure 13D shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (146). Aliquots of 100 μL were taken at 0, 5, 30, 90, and 150 min and analyzed by LC-MS. [Figure 13E]Figure 13E shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((6aS)-3-(4-((5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutoxy)-6-hydroxy-2-methoxy-12-oxo-5,6,6a,7,8,9,10,12-octahydrobenzo[e]pyrido[1,2-a][1,4]diazepine-5-carbonyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (146). Aliquots of 100 μL were taken at 0, 5, 30, 90, and 150 min and analyzed by LC-MS. [Figure 14A] Figure 14A shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((5-(3-carboxypropoxy)-4-methoxy-2-((S)-2-((methoxyimino)methyl)piperidine-1-carbonyl)phenyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (150). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 14B] Figure 14B shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((5-(3-carboxypropoxy)-4-methoxy-2-((S)-2-((methoxyimino)methyl)piperidine-1-carbonyl)phenyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (150). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 14C]Figure 14C shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((5-(3-carboxypropoxy)-4-methoxy-2-((S)-2-((methoxyimino)methyl)piperidine-1-carbonyl)phenyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (150). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 14D] Figure 14D shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((5-(3-carboxypropoxy)-4-methoxy-2-((S)-2-((methoxyimino)methyl)piperidine-1-carbonyl)phenyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (150). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 14E] Figure 14E shows the results of a β-glucuronidase assay using (2S,3S,4S,5R,6S)-6-(4-((((5-(3-carboxypropoxy)-4-methoxy-2-((S)-2-((methoxyimino)methyl)piperidine-1-carbonyl)phenyl)carbamoyl)oxy)methyl)-2-nitrophenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (150). 100 μL aliquots were taken at 0, 5, 30, 90, and 150 minutes and analyzed by LC-MS. [Figure 15] FIG. 15 shows the HIC profile of trastuzumab. [Figure 16] Figure 16 shows the PLRP trace of trastuzumab, with heavy chain (left peak) and light chain (right peak) peaks as indicated. [Figure 17]Figure 17 shows the HIC profile of trastuzumab-91. The average DAR was calculated as 8. The conjugation process did not result in significant aggregation compared to the starting antibody, with 93.1% monomer formed. [Figure 18] Figure 18 shows the SEC profile of trastuzumab-91, which is 93.1% monomeric. No free toxin linker was detectable in the ADC sample. [Figure 19] FIG. 19 is a graph showing the dose tolerability of trastuzumab-91 (DAR8) in non-tumor-bearing CD1 mice. [Figure 20] FIG. 20 is a graph showing the in vivo efficacy of trastuzumab-91 (DAR8) at a single dose of 5 mg / kg. [Figure 21] FIG. 21 shows the protection of the N11-C12-alkylated imine position or the masking of the C8-position of the G-alkylated PDD-construct using a sugar-based moiety. [Figure 22] FIG. 22 is a scheme showing the PDD-prodrug compounds before and after cleavage of the sugar group. [Figure 23] FIG. 23 is a graph showing the in vivo efficacy of Compound 180 at a single dose of 5 mg / kg. DETAILED DESCRIPTION OF THE INVENTION

[0058] 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 disclosure belongs. All patents and publications mentioned herein are incorporated by reference in their entirety.

[0059] definition The following abbreviations are used throughout: Ac acetyl, Alloc allyloxycarbonyl, Boc tert-butyloxycarbonyl, DHP dihydropyran, DMAP 4-dimethylaminopyridine, DMF dimethylformamide, EDCI 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide, Et ethyl, Me methyl, Ph phenyl, Tf trifluoromethanesulfonate, TFA trifluoroacetic acid, THF tetrahydrofuran, and THP tetrahydropyranyl.

[0060] "Substituted," when used in connection with a chemical substituent or moiety (e.g., an alkyl group), means that one or more hydrogen atoms of the substituent or moiety have been replaced with one or more non-hydrogen atoms or groups, provided that valency requirements are met and the substitution results in a chemically stable compound.

[0061] "Optionally substituted" refers to a parent group that can be unsubstituted or substituted with one or more substituents. Preferably, unless otherwise specified, an optionally substituted parent group contains 1 to 3 optional substituents, if optional substituents are present. When a group can be "optionally substituted with 1, 2, or 3 groups," this means that the group can be substituted with 0, 1, 2, or 3 optional substituents. Preferably, the group is substituted with 1, 2, or 3 optional substituents. When a group can be "optionally substituted with one or two optional substituents," this means that the group can be substituted with 0, 1, or 2 of the optional substituents. Preferably, the group can be substituted with 0 or 1 optional substituent. In some embodiments, preferably, the group is unsubstituted. In other embodiments, preferably, the group is substituted with one of the optional substituents.

[0062] The optional substituents are C 1-8 Alkyl, C 2-7 Alkenyl, C 2-7 Alkynyl, C 1-12 Alkoxy, C 5-20 Aryl, C3-10 Cycloalkyl, C 3-10 Cycloalkenyl, C 3-10 Cycloalkynyl, C 3-20 Heterocyclyl, C 3-20 The optional substituents may be selected from heteroaryl, acetal, acyl, acylamido, acyloxy, amidino, amido, amino, aminocarbonyloxy, azido, carboxy, cyano, ether, formyl, guanidino, halo, hemiacetal, hemiketal, hydroxamic acid, hydroxyl, imidic acid, imino, ketal, nitro, nitroso, oxo, oxycarbonyl, oxycarboyloxy, sulfamino, sulfamyl, sulfate ester, sulfhydryl, sulfinamino, sulfinate ester, sulfino, sulfinyl, sulfinyloxy, sulfo, sulfonamido, sulfonamino, sulfonate ester, sulfonyl, sulfonyloxy, and uredio groups. In some embodiments, the optional substituents are selected from OH, C 1-8 Alkyl, OC 1-12 and 1, 2, or 3 optional substituents independently selected from alkyl, OH, C 1-8 Alkyl, and OC 1-12 alkyl, more preferably the optional substituents are selected from C 1-8 Alkyl and OC 1-12 alkyl.

[0063] "Independently" or "independently selected" means, for example, "each R', R'' is independently selected from H, C 1-8 Used in the context of a statement such as "is alkyl," this means that each instance of a functional group, e.g., R', is selected from the listed alternatives independently of any other instances of R" or R" in the compound. Thus, for example, for the first instance of R' in the compound, H may be selected, for the next instance of R' in the compound, methyl may be selected, and for the first instance of R" in the compound, ethyl may be selected.

[0064] "C 1-8"Alkyl" refers to straight-chain and branched saturated hydrocarbon groups generally having from 1 to 8 carbon atoms, C 1-7 Alkyl is preferred, C 1-6 Alkyl is preferred, C 1-5 Alkyl is preferred, C 1-4 Alkyl is more preferred, C 1-3 Examples of alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, pent-1-yl, pent-2-yl, pent-3-yl, 3-methylbut-1-yl, 3-methylbut-2-yl, 2-methylbut-2-yl, 2,2,2-trimethyleth-1-yl, n-hexyl, n-heptyl, and n-octyl.

[0065] "Alkylene" refers to a divalent radical derived from an alkane which can be straight-chained or branched, as exemplified by -CH2CH2CH2CH2-. The alkylene can have the number of carbon atoms described above for an alkyl group.

[0066] The term "amino acid" refers to naturally occurring (or "canonical") α-amino acids and their stereoisomers, unnatural (or "non-canonical") amino acids and their stereoisomers, as well as modified or synthetic amino acids that function in a manner similar to the naturally occurring amino acids, and amino acid analogs and amino acid mimetics. An "stereoisomer" of an amino acid refers to an enantiomer of an amino acid, such as an L-amino acid or a D-amino acid. For example, a stereoisomer of a naturally occurring amino acid refers to an enantiomer of a naturally occurring amino acid, i.e., a D-amino acid.

[0067] Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are subsequently modified, such as hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Naturally occurring α-amino acids include, but are not limited to, amino acids selected from alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val). Stereoisomers of naturally occurring α-amino acids include D-alanine (D-Ala), D-arginine (D-Arg), D-asparagine (D-Asn), D-aspartic acid (D-Asp), D-cysteine ​​(D-Cys), D-glutamine (D-Gln), D-glutamic acid (D-Glu), D-glycine (D-Gly), D-histidine (D-His), and D-isoleucine (D-I). Examples of amino acids include, but are not limited to, D-le), D-leucine (D-Leu), D-lysine (D-Lys), D-methionine (D-Met), D-phenylalanine (D-Phe), D-proline (D-Pro), D-serine (D-Ser), D-threonine (Thr), D-tryptophan (D-Trp), D-tyrosine (D-Tyr), and D-valine (D-Val).

[0068] Non-natural amino acids include, but are not limited to, either L- or D-amino acid analogs, amino acid mimetics, and synthetic amino acids that function in a manner similar to natural amino acids. For example, an "amino acid analog" is a non-natural amino acid that has the same basic chemical structure as a natural amino acid, i.e., an α-carbon bonded to a hydrogen atom, a carboxyl group, an amino group, and an R group, such as homoserine, norleucine, methionine sulfoxide, and methionine methylsulfonium. Such analogs may have modified R groups (e.g., norleucine) or modified peptide backbones, but may retain the same basic chemical structure as a natural amino acid.

[0069] Non-limiting examples of unnatural amino acids include 1-aminocyclopentane-1-carboxylic acid (Acp), 1-aminocyclobutane-1-carboxylic acid (Acb), 1-aminocyclopropane-1-carboxylic acid (Acpc), citrulline (Cit), homocitrulline (HoCit), α-aminohexanedioic acid (Aad), 3-(4-pyridyl)alanine (4-Pal), 3-(3-pyridyl)alanine (3-Pal), propargylglycine (Pra), α-aminoisobutyric acid (Aib), α-aminobutyric acid (Abu), and the like. ), norvaline (Nva), α,β-diaminopropionic acid (Dpr), α,γ-diaminobutyric acid (Dbu), α-tert-butylglycine (Bug), 3,5-dinitrotyrosine (Tyr(3,5-diNO2)), norleucine (Nle), 3-(2-naphthyl)alanine (Nal-2), 3-(1-naphthyl)alanine (Nal-1), cyclohexylalanine (Cha), di-n-propylglycine (Dpg), cyclopropylalanine (Cpa), homoleucine (Hle), homoserine (HoS er), homoarginine (Har), homocysteine ​​(Hcy), methionine sulfoxide (Met(O)), methionine methylsulfonium (Met(S-Me)), α-cyclohexylglycine (Chg), 3-benzo-thienylalanine (Bta), taurine (Tau), hydroxyproline (Hyp), O-benzyl-hydroxyproline (Hyp(Bzl)), homoproline (HoPro), β-homoproline (βHoPro), thiazolidine-4-carboxylic acid (Thz), nipecotic acid (N ip), isonipecotic acid (IsoNip), 3-carboxymethyl-1-phenyl-1,3,8-triazaspiro[4,5]decan-4-one (Cptd), tetrahydro-isoquinoline-3-carboxylic acid (3-Tic), 5H-thiazolo[3,2-a]pyridine-3-carboxylic acid (Btd), 3-aminobenzoic acid (3-Abz), 3-(2-thienyl)alanine (2-Thi), 3-(3-thienyl)alanine (3-Thi), α-aminooctanedioc acid (Asu), diethylglycine (Deg), 4-amino-4-carboxy-1,1-Dioxo-tetrahydrothiopyran (Acdt), 1-amino-1-(4-hydroxycyclohexyl)carboxylic acid (Ahch), 1-amino-1-(4-ketocyclohexyl)carboxylic acid (Akch), 4-amino-4-carboxytetrahydropyran (Actp), 3-nitrotyrosine (Tyr(3-NO2)), 1-amino-1-cyclohexanecarboxylic acid (Ach), 1-amino-1-(3-piperidinyl)carboxylic acid (3-Apc), 1-amino-1-(4-piperidinyl)carboxylic acid (4-Apc), 2-amino-3-(4-piperidinyl)propionic acid (4-App), 2-aminoindan-2-carboxylic acid (Aic), 2-amino-2-naphthylacetic acid (Ana), (2S,5R)-5-phenylpyrrolidine Examples of phenylalanine analogs include phenylalanine-2-carboxylic acid (Ppca), 4-thiazolylalanine (Tha), 2-aminooctanoic acid (Aoa), 2-aminoheptanoic acid (Aha), ornithine (Orn), azetidine-2-carboxylic acid (Aca), α-amino-3-chloro-4,5-dihydro-5-isoazoleacetic acid (Acdi), thiazolidine-2-carboxylic acid (Thz(2-COOH)), allylglycine (Agl), 4-cyano-2-aminobutyric acid (Cab), 2-pyridylalanine (2-Pal), 2-quinoylalanine (2-Qal), cyclobutylalanine (Cba), phenylalanine analogs, derivatives of lysine, ornithine (Orn) and α,γ-diaminobutyric acid (Dbu), stereoisomers thereof, and combinations thereof (e.g., Liu (See, e.g., E. et al., Anal. Biochem., 295:9-16 (2001)). Thus, unnatural α-amino acids exist as either unnatural L-α-amino acids, unnatural D-α-amino acids, or combinations thereof.

[0070] An "amino acid mimetic" is a chemical compound that has a structure different from the normal chemical structure of an amino acid, but functions in a manner similar to a natural amino acid. Suitable amino acid mimics include, but are not limited to, β-amino acids and γ-amino acids. In β-amino acids, the amino group is bonded to the β-carbon atom of the carboxyl group so that there are two carbon atoms between the amino group and the carboxyl group. In γ-amino acids, the amino group is bonded to the γ-carbon atom of the carboxyl group so that there are three carbon atoms between the amino group and the carboxyl group. Suitable R groups for β-amino acids or γ-amino acids include, but are not limited to, the side chains found in natural and unnatural amino acids.

[0071] "C 6-26 "Aralkyl" refers to an arylalkyl group having 6 to 26 carbon atoms, including an alkyl group substituted with an aryl group. Preferably, the alkyl group is C 1-6 The alkyl group is phenyl, and the aryl group is phenyl. 6-26 Examples of aralkyl include benzyl and phenethyl. In some cases, C 6-26 The aralkyl group may be optionally substituted, and may be an optionally substituted C 6-26 An example of an aralkyl group is 4-methoxybenzyl.

[0072] "C 5-20 "Aryl" refers to fully unsaturated monocyclic, bicyclic, and polycyclic aromatic hydrocarbons having at least one aromatic ring and a specified number of carbon atoms including those ring members (e.g., C 5-20 Aryl refers to an aryl group having 5 to 20 carbon atoms as ring members. An aryl group may be attached to a parent group or substrate at any ring atom and may contain one or more non-hydrogen substituents unless such attachment or substitution violates valence requirements. Preferably, C 6-14 Aryl is C 6-12 Aryl, more preferably C 6-10 Examples of aryl groups include phenyl.

[0073] "Arylene" refers to a divalent radical derived from an aryl group, for example, —C 6 H 4 —, an arylene derived from phenyl.

[0074] "C 3-8 "Cycloalkyl" or "3 to 8-membered cycloalkyl" means a closed ring of carbon atoms having 3 to 8 carbon atoms, preferably 3 to 7 carbon atoms, more preferably 3 to 6 carbon atoms, and includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0075] "C 3-8 "Cycloalkylene" or "3- to 8-membered cycloalkylene" refers to a divalent radical derived from a cycloalkyl group, e.g., —CH 10 -It is.

[0076] "C 3-8 "Cycloalkenylene" refers to a cycloalkenyl group, i.e., a divalent radical derived from a carbocyclic group having one or more C=C, e.g., -C6H8-.

[0077] Halogen or halo refers to a group selected from F, Cl, Br, and I. Preferably, halogen or halo is F or Cl. In some embodiments, preferably, halogen is F. In other embodiments, preferably, halogen is Cl.

[0078] "C 5-10"Heteroaryl" or "5- to 10-membered heteroaryl" refers to an unsaturated monocyclic or bicyclic aromatic group containing 5 to 10 ring atoms, 1 to 5 of which are ring heteroatoms, whether carbon or heteroatoms. Preferably, any monocyclic heteroaryl ring has 5 to 6 ring atoms and 1 to 3 ring heteroatoms. Preferably, each ring heteroatom is independently selected from nitrogen, oxygen, and sulfur. Bicyclic rings include fused ring systems, particularly bicyclic groups in which a monocyclic heterocycle containing 5 ring atoms is fused to a benzene ring. A heteroaryl group may be attached to a parent group or substrate at any ring atom and may contain one or more non-hydrogen substituents unless such attachment or substitution would violate valence requirements or result in a chemically unstable compound.

[0079] Examples of monocyclic heteroaryl groups include, but are not limited to, those derived from: N1: pyrrole, pyridine, O1: Fran, S1: thiophene, N1O1: oxazole, isoxazole, isoxazine, N2O1: oxadiazole (e.g., 1-oxa-2,3-diazolyl, 1-oxa-2,4-diazolyl, 1-oxa-2,5-diazolyl, 1-oxa-3,4-diazolyl), N3O1: oxatriazole, N1S1: Thiazole, isothiazole, N2: Imidazole, pyrazole, pyridazine, pyrimidine, pyrazine, N3: Triazoles, triazines, and N4: tetrazole.

[0080] Examples of heteroaryls containing fused rings include, but are not limited to, those derived from: O1: benzofuran, isobenzofuran, N1: indole, isoindole, indolizine, isoindoline, S1: benzothiofuran, N1O1: benzoxazole, benzisoxazole, N1S1: benzothiazole, N2: benzimidazole, indazole, O2: benzodioxole, N2O1: benzofurazan, N2S1: benzothiadiazole, N3: benzotriazole, and N4: purines (e.g., adenine, guanine), pteridines, "Heteroarylene" refers to a divalent radical derived from a heteroaryl group (such as those described above), exemplified by pyridinyl-[CHN]-. Heteroarylene can be a monocyclic, bicyclic, or tricyclic ring system. Representative heteroarylenes include, but are not limited to, triazolylene, tetrazolylene, oxadiazolylene, pyridylene, furylene, benzofuranylene, thiophenylene, benzothiophenylene, quinolinylene, pyrrolylene, indolylene, oxazolylene, benzoxazolylene, imidazolylene, benzimidazolylene, thiazolylene, benzothiazolinylene, and the like. Heteroarylene may be selected from aryl, isoxazolylene, pyrazolylene, isothiazolylene, pyridazinylene, pyrimidinylene, pyrazinylene, triazinylene, cinnolinylene, phthalazinylene, quinazolinylene, pyrimidylene, azepinylene, oxepinylene, and quinoxalinylene. Heteroarylene may be optionally substituted.

[0081] "C6-16 "Heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group. Preferably, the alkyl is a C 1-6 alkyl groups and heteroaryl groups as defined above. 5-10 Heteroaryl. C 6-16 Examples of heteroarylalkyl groups include pyrrol-2-ylmethyl, pyrrol-3-ylmethyl, pyrrol-4-ylmethyl, pyrrol-3-ylethyl, pyrrol-4-ylethyl, imidazol-2-ylmethyl, imidazol-4-ylmethyl, imidazol-4-ylethyl, thiophen-3-ylmethyl, furan-3-ylmethyl, pyridin-2-ylmethyl, pyridin-2-ylethyl, thiazol-2-ylmethyl, thiazol-4-ylmethyl, thiazol-2-ylethyl, pyrimidin-2-ylpropyl, and the like.

[0082] "C 3-20 "Heterocyclyl" refers to a saturated or partially unsaturated monocyclic, bicyclic, or polycyclic group having from 3 to 20 ring atoms, of which from 1 to 10 are ring heteroatoms, whether carbon or heteroatoms. Preferably, each ring has from 3 to 8 ring atoms and 1 to 4 ring heteroatoms (e.g., preferably C 3-5 Heterocyclyl refers to a heterocyclyl group having 3 to 5 ring atoms and 1 to 4 heteroatoms as ring members.) The ring heteroatoms are independently selected from nitrogen, oxygen, and sulfur.

[0083] Like bicyclic cycloalkyl groups, bicyclic heterocyclyl groups can include isolated rings, spirocyclic rings, fused rings, and bridged rings. A heterocyclyl group can be attached to a parent group or substrate at any ring atom and can contain one or more non-hydrogen substituents unless such attachment or substitution violates valence requirements or results in a chemically unstable compound.

[0084] Examples of monocyclic heterocyclyl groups include, but are not limited to, those derived from: N1: aziridine, azetidine, pyrrolidine, pyrroline, 2H-pyrrole or 3H-pyrrole, piperidine, dihydropyridine, tetrahydropyridine, azepine, O1: oxirane, oxetane, tetrahydrofuran, dihydrofuran, tetrahydropyran, dihydropyran, pyran, oxepin, S1: Thiirane, thietane, tetrahydrothiophene, tetrahydrothiopyran, thiepane, O2: dioxoiane, dioxane, and dioxepane, O3: Trioxane, N2: Imidazoiidine, pyrazolidine, imidazoline, pyrazoline, piperazine, N1O1: tetrahydrooxazole, dihydrooxazole, tetrahydroisoxazole, dihydroisoxazole, morpholine, tetrahydrooxazine, dihydrooxazine, oxazine, N1S1: Thiazoline, thiazolidine, thiomorpholine, N2O1: oxadiazine, O1S1: Oxathioles and oxathianes (thioxanes), and N1O1S1: Oxathiazine.

[0085] Examples of substituted monocyclic heterocyclyl groups include heterocyclyl groups derived from cyclic forms of sugars, such as furanoses, such as arabinofuranose, lyxofuranose, ribofuranose, and xylofuranse, and pyranoses, such as aliopyranose, altropyranose, glucopyranose, mannopyranose, gulopyranose, idopyranose, galactopyranose, and talopyranose.

[0086] "Nucleic acid" refers to a linear polymer of nucleosides (including deoxyribonucleosides, ribonucleosides, or analogs thereof) linked by internucleoside linkages. Nucleic acid can encompass the terms "polynucleotide" and "oligonucleotide." A linear polymer can be represented by a sequence of letters such as "ATGCCTG," where, unless otherwise noted, the nucleotides are in 5' to 3' order from left to right, and "A" represents deoxyadenosine, "C" represents deoxycytidine, "G" represents deoxyguanosine, and "T" represents deoxythymidine. Another naturally occurring nucleotide is "U," which represents uridine. The letters A, C, G, T, and U can be used, as is standard in the art, to refer to the base itself, a nucleoside, or a nucleotide containing the base. In naturally occurring nucleic acids, the internucleoside linkages are typically phosphodiester bonds, and the subunits are referred to as "nucleotides." Nucleic acids can also contain other internucleoside linkages, such as phosphoro-thioate linkages. Such analogs of nucleotides that do not contain a phosphate group are considered to be within the scope of the term "nucleotide" as used herein, and nucleic acids that contain one or more internucleoside linkages that are not phosphodiester linkages will still be referred to as "polynucleotides," "oligonucleotides," etc.

[0087] Nitrogen Protecting Groups Nitrogen protecting groups are well known in the art and are groups that block or protect nitrogen groups from further reaction.Nitrogen protecting groups are exemplified by carbamates, such as methyl or ethyl carbamates, 9-fluorenylmethyloxy-carbonyl (Fmoc), substituted ethyl carbamates, carbamates cleaved by 1,6-beta elimination, urea, amide, peptide, alkyl, and aryl derivatives.Carbamate protecting groups have the following general formula: [ka] In this specification, a zigzag line (or a wavy line) [ka] indicates the point of attachment of the indicated group (e.g., a protecting group as described above) to the remainder of the compound of formula (I). Suitable nitrogen protecting groups may be selected from acetyl, trifluoroacetyl, t-butyloxy-carbonyl (BOC), benzyloxycarbonyl (Cbz), and 9-fluorenylmethyloxy-carbonyl (Fmoc).

[0088] A number of possible carbamate nitrogen protecting groups are described in Wuts, P.G.M. and Greene, T.W., Protective Groups in Organic Synthesis, 4 th Edition, Wiley-Interscience, 2007, pp. 706-771, and P. Kocienski, Protective Groups, 3rd Edition (2005), which are incorporated herein by reference.

[0089] Particularly preferred protecting groups include Alloc (allyloxycarbonyl), Troc (2,2,2-trichloroethyl carbonate), Teoc [2-(trimethylsilyl)ethoxycarbonyl], BOC (tert-butyloxycarbonyl), Doc (2,4-dimethylpent-3-yloxycarbonyl), Hoc (cyclohexyloxy-carbonyl), TcBOC (2,2,2-trichloro-tert-butyloxycarbonyl), Fmoc (9-fluorenylmethyloxycarbonyl), 1-Adoc (1-adamantyloxycarbonyl), and 2-Adoc (2-adamantyloxycarbonyl).

[0090] Hydroxyl Protecting Groups Hydroxyl protecting groups are well known in the art, and many suitable groups are described in Wuts, P.G.M. and Greene, T.W., Protective Groups in Organic Synthesis, 4 thEdition, Wiley-Interscience, 2007, pp. 16-366, and P. Kocienski, Protective Groups, 3rd Edition (2005), which are incorporated herein by reference.

[0091] Classes of particular interest include silyl ethers, methyl ethers, alkyl ethers, benzyl ethers, esters, benzoates, carbonates, and sulfonates.

[0092] Suitable protecting groups include THP (tetrahydropyranyl ether). Hydroxyl can also be protected as N-methylpiperazine carbamate, especially the hydroxyl group of a phenol.

[0093] L S In an embodiment, L S is R S In some embodiments, L is a bond or linker moiety that is bonded or connected to S bond, amino acids, peptide chains having 2 to 6 amino acids, alkylene chains containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, paraformaldehyde chains -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are P, O, S, NH, C 5-9 which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted. 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be selected from one, two, or three independently selected optional R 20 It may be substituted with a group.

[0094] In some embodiments, L S L2, R 28 , or L2-R 28 Includes.

[0095] In some embodiments, the linker L s comprises one or more groups selected from sulfamino, sulfamyl, sulfate, sulfhydryl, sulfinamino, sulfinate, sulfino, sulfinyl, sulfinyloxy, sulfo, sulfonamido, sulfonamino, sulfonate, sulfonyl, sulfonyloxy, phosphate, phosphoramidate, thiophosphate, phosphonate, and thiophosphonate.

[0096] In some embodiments, L S comprises a benzyl carboxylate group. In some embodiments, L S includes the following: [ka] where v is 0, 1, 2, or 3, and each R 40 are independently -NO2, -SO3R 26 , -C(=O)-R 26 , -C(=O)-Cl, -C 1‐6 Fluoroalkyl, -C 1-6 Fluoroalkoxy, R 20 , R 28 , -[CH2CH2O] 1-50 -R 28 , and -NH-X AA -R 28 wherein X AA is an amino acid sequence having 1 to 20 amino acid portions. In some embodiments, v is 1, 2, or 3, and each R 40 are independently -NO2, -SO3R 26 , -C(=O)-R 26 , -C(=O)-Cl, -C 1-6 Fluoroalkyl, -C 1-6Fluoroalkoxy, -F, -OH, -NH2, -CN, -NCO, -(CH2) j -CO2R 26 , -C(=O)-NR 26 R 27 , L2-R 28 , maleimide, [ka] wherein X is selected from AA is an amino acid sequence having 1 to 10 amino acid moieties.

[0097] In some embodiments, L S are as follows: [ka]

[0098] R S In some embodiments, the carbohydrate substituent of the compound of Formula (I) is R S In some embodiments, R S is glycosyl or O-glycosyl.

[0099] "Saccharide" refers to a sugar which may be a monosaccharide or a disaccharide; S" refers to a monovalent substituent of a monosaccharide and / or disaccharide derived from a monosaccharide and / or disaccharide. The sugar may suitably be a pentose or hexose, or a disaccharide containing a pentose and / or hexose. Examples of monosaccharides include glucose, fructose, galactose, ribose, ribulose, and stereoisomers of these sugars. As used herein, sugar refers to the furanose, acyclic, and / or pyranose forms of sugars, and a formula showing one form of sugar is also intended to include other forms of sugars unless the context requires otherwise. A monovalent substituent sugar may contain the glycosyl group of the corresponding monosaccharide (or disaccharide) that can be obtained by removing a hydroxyl group from a monosaccharide or disaccharide or by removing a hydrogen from a hydroxyl group of a monosaccharide or disaccharide. Non-limiting examples of furanose include arabinofuranose, lyxofuranose, ribofuranose, xylofuranse, and the like. Non-limiting examples of pyranose include ariopyranose, altropyranose, glucopyranose, mannopyranose, gulopyranose, idopyranose, galactopyranose, talopyranose, and the like.

[0100] R T In embodiments, R T is L2-R 28 , phenyl, and C 5-9 heteroaryl, selected from phenyl and C 5-9 A heteroaryl group may be substituted with up to three optional substituents. Thus, R T Phenyl group or C selected 5-9 Any of the heteroaryl groups may be optionally substituted with up to three optional substituents.

[0101] Preferably, R T is L2-R 28, phenyl, pyrrolyl, N-methylpyrrolyl, furanyl, thiophenyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzoxazolyl and benzothiazolyl, wherein the phenyl, pyrrolyl, N-methylpyrrolyl, furanyl, thiophenyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, pyridyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzoxazolyl and benzothiazolyl groups are selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, L2-R 28 , (CH2) j -CO2R 11 , O-(CH2) k -NR 11 R 12 , (CH2) j -NR 11 R 12 , C(=O)-NH-(CH2) k -NR 11 R 12 , C(=O)-NH-R 24 and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 and optionally substituted with up to three optional substituents selected from:

[0102] Preferably, R T is L2-R 28, phenyl, pyrrolyl, N-methylpyrrolyl, furanyl, thiophenyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzoxazolyl and benzothiazolyl, and the phenyl, pyrrolyl, N-methylpyrrolyl, furanyl, thiophenyl, imidazolyl, N-methylimidazolyl, oxazolyl, thiazolyl, benzofuranyl, benzothiophenyl, benzimidazolyl, N-methylbenzimidazolyl, benzoxazolyl and benzothiazolyl groups are selected from OH, C 1-6 Alkyl, OC 1-6 Alkyl, L2-R 28 , (CH2) j -CO2R 11 , O-(CH2) k -NR 11 R 12 , (CH2) j -NR 11 R 12 , C(=O)-NH-(CH2) k -NR 11 R 12 , C(=O)-NH-R 24 and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 and optionally substituted with one or two optional substituents selected from:

[0103] Preferably, R T is L2-R 28 , phenyl, N-methylpyrrolyl, thiophenyl, N-methylimidazolyl, oxazolyl, thiazolyl, benzothiophenyl, N-methylbenzimidazolyl and benzothiazolyl, wherein the phenyl, N-methylpyrrolyl, thiophenyl, N-methylimidazolyl, oxazolyl, thiazolyl, benzothiophenyl, N-methylbenzimidazolyl and benzothiazolyl groups are selected from the group consisting of OH, C 1-6 Alkyl, OC 1-6 Alkyl, L2-R 28 , (CH2) j -CO2R 11O-(CH2) k -NR 11 R 12 , (CH2) j -NR 11 R 12 , C(=O)-NH-(CH2) k -NR 11 R 12 , C(=O)-NH-R 24 and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 and optionally substituted with one or two optional substituents selected from:

[0104] Preferably, R T OH, C 1-6 Alkyl, OC 1-6 Alkyl, L2-R 28 , (CH2) j -CO2R 11 , O-(CH2) k -NH2, (CH2) j -NH2, C(=O)-NH-(CH2) k -NH2, C(=O)-NH-R 24 and C(=O)-NH-(CH2) k and -C(=NH)NH2.

[0105] Preferably, R T is an optionally substituted C(=O)-NH-R 24 where R 24 is -C6H4-(CH2) j -R 18 and the phenylene group -C6H4- is substituted at the para position.

[0106] Preferably, R T is OH, methyl, ethyl, OCH3, OCH2CH3, L2-R 28 , CO2H, CO2CH3, CO2CH2CH3, O-(CH2) k -NH2, and (CH2) j -NH2.

[0107] Preferably, R T may be substituted with one or two optional substituents.

[0108] More preferably, R T may be substituted with one optional substituent.

[0109] More preferably, R T is selected from the following: [ka] In the formula, Z1 is NH, N-CH3, NR S , S and O; Z2 is selected from CH and N; Z3 is selected from S and O; Z4, CH, CR S , and N, R 22 -L2-R 28 , (CH2) j CO2R 11 , (CH2) j NR 11 R 12 , and C(=O)-NH-C6H4-(CH2) j -R 18 is selected from R 18 -L2-R 28 , CO2R 11 , and N.R. 11 R 12 is selected from j is selected from an integer from 0 to 6; R 11 and R 12 are independently H, -L2-R 28 , and C 1-6 alkyl, R 23 H, R S , -L2-R 28 , and C 1-6 alkyl.

[0110] The wavy line represents the R T indicates the point of attachment of the group to the remainder of the compound of formula (I).

[0111] More preferably, R T is selected from the following: [ka] In the formula, Z1 is NH, N-CH3, NR S , S and O; Z2 is selected from CH and N; Z3 is selected from S and O; Z4 is CH, R S , and N, R 11 H, L2-R 28 , and C 1-6 alkyl, R 23 H, R S , L2-R 28 , and C 1-6 alkyl.

[0112] L 2 In embodiments, linker L2 is a bond or a moiety having 1 to 200 non-hydrogen atoms selected from C, N, P, O, S, or halogen, and may incorporate an ether, oxo, carboxyl, carboxamido, carboxamidyl, urethanyl, branched, cyclic, unsaturated, amino acid, heterocyclyl, aryl, or heteroaryl moiety. In embodiments, linker L2 is a bond or a moiety selected from an amino acid, a peptide chain having 2 to 100 amino acids, an alkylene chain containing 1 to 50 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-50 - and polyethylene glycol chain -(OCH2CH2) 1-50 -, and these chains contain one or more groups selected from P, O, S, NH, C 5-9may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; C 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted. In some embodiments, peptide, alkylene, paraformaldehyde, and polyethylene glycol chains, and / or C 5-9 Heteroarylene, phenylene, heterocyclyl, and cycloalkyl moieties can be selected from one, two, or three independently selected optional R 20 Groups and / or L s -R s It may be substituted with a group.

[0113] Linker L2 can be unbranched or branched, flexible or rigid, short or long, and can incorporate any combination of moieties deemed useful. In some embodiments, at least a portion of linker L2 can have a polyalkylene oxide polymer region, which can improve the solubility of compounds of Formula (I) or (II). In some embodiments, linker L2 can have repeating ethylene glycol units, and can have a number of repeating ethylene glycol units ranging from about 1 to about 25, or any number therebetween. In some embodiments, L2 can contain about 3 to about 20, about 4 to about 15, about 5 to about 12, or about 6 to about 10 ethylene glycol units. In some embodiments, at least a portion of linker L2 can contain one or more amino acid moieties that can provide improved solubility for compounds of Formula (I) or (II) or that can provide an amino acid sequence that enhances target binding, improving compatibility with targeting agents, or improving target binding recognition. In other embodiments, linker L2 can include one or more amino acid moieties that provide a suitable substrate motif for a protease. When a set of amino acid moieties is incorporated into linker L2 that provides a substrate motif specific to a selected protease, the cytotoxic drug compound of Formula (I) or (II) can be released from the target-bound complex to provide a localized cytotoxic effect. Such substrate motifs are known in the art and can be incorporated into linker L2 as needed to provide selective release from the target-bound complex. This selectivity can be based on the known presence of the desired protease within the localized delivery region of the conjugated drug. Other polymer-type moieties, such as polyacids, polysaccharides, or polyamines, can be incorporated into linker L2. Other moieties, such as substituted aromatic or heteroaromatic moieties, can be used to increase rigidity or provide synthetically accessible sites on substituents within the moiety for attachment to reactive moieties or G, Q, or T groups of compounds of Formula (I).

[0114] In some embodiments, the linker L2 may comprise one or more groups selected from sulfamino, sulfamyl, sulfate, sulfhydryl, sulfinamino, sulfinate, sulfino, sulfinyl, sulfinyloxy, sulfo, sulfonamido, sulfonamino, sulfonate, sulfonyl, sulfonyloxy, phosphate, phosphoramidate, thiophosphate, phosphonate, and thiophosphonate.

[0115] For example, the linker L2 may comprise ethylene glycol repeat units and / or an amino acid sequence. In some embodiments, the linker L2 comprises the following formula: -[CH2CH2O] 0-50 -X AA - In the formula, X AA is the amino acid sequence.

[0116] In some embodiments, the linker L2 comprises or consists of the following formula: [ka] In the formula, R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl; L C is an amino acid, an amino acid derivative, a peptide chain having 2 to 6 amino acids or amino acid derivatives, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-8 -, and these chains contain one or more groups selected from one or more P, O, S and / or NH groups and / or C 5-9 may be interrupted by heteroarylene and / or phenylene, and each C 5-9 The heteroarylene group and / or each phenylene group may be substituted. In embodiments, L C is R 28is further complexed to

[0117] In some embodiments, the linker L2 comprises or consists of the following formula: [ka]

[0118] In some embodiments, L C is the expression [ka] at will [ka] It has.

[0119] In some embodiments, R 28 -L C comprises a maleimide, optionally the maleimide is of the formula [ka] at will [ka] It has.

[0120] In some embodiments, L C is a polyethylene glycol chain -(OCH2CH2) 1-10 -, optionally including -(OCH2CH2)8-.

[0121] In some embodiments, L C is selected from the following: [ka]

[0122] In some embodiments, R 28 -L C is selected from the following: [ka]

[0123] In some embodiments, R 28 L2 comprises or consists of the following formula: [ka] During the ceremony R 28 are azides, alkynes, bisulfones, carbohydrazides, hydrazines, hydroxylamines, iodoacetamides, isothiocyanates, maleimides, phosphines, pyridopyridazines, semihydrazides, succinimidyl esters, sulfodichlorophenol esters, sulfonyl halides, sulfosuccinimidyl esters, 4-sulfotetrafluorophenyl esters, tetrafluorophenyl esters, thiazoles, R A , O-(CH2) k -NR 26 R 26 , NHNH2, or a targeting agent, wherein the targeting agent is selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; L C is the expression [ka] wherein X 10 is a single bond, -HN-[CH2-CH2-O] p -(CH2) 1-5 -C(O)-, -HN-[CH2-CH2-O] p -(CH2) 1-5 -C(O)-NH-, or [ka] wherein p is independently at each occurrence an integer from 1 to 50, and Alk is C1-C5 alkyl; X 20 is a single bond, [ka] In some embodiments, L C is the expression [ka] In some embodiments, L C is the expression [ka] In some embodiments, p is an integer between 5 and 10, for example, 5, 6, 7, 8, 9, or 10. In some embodiments, p is 8. In some embodiments, X 10 is -HN-[CH2-CH2-O] p In some embodiments, X is —(CH)—C(O)—. 10 is a single bond. 10 is -HN-[CH2-CH2-O] p In some embodiments, X is —(CH)—C(O)—N—. 10 teeth, [ka] In some embodiments, Alk is -CH3. In some embodiments, X 20 is a single bond. 20 teeth, [ka] In some embodiments, X 20 teeth, [ka] is.

[0124] Any suitable number of ethylene glycol units can be used in the linker L2 of the present disclosure. For example, the linker L2 can include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 16, 19, 20, 23, 24, 35, 36, 37, 48, 49, or more ethylene glycol units. In some embodiments, the linker L2 can include 8 ethylene glycol units. Some commercially available ethylene glycol groups (polyethylene glycol, PEG) having monodisperse ("d") polyethylene glycol with 8 ethylene glycol repeat units are suitable for the linker L2, such as H2N-PEG8-C(O)OH. Other monodisperse PEG units are commercially available, for example, from Advanced ChemTech, and are known to those skilled in the art. In some embodiments, the linker L2 includes the following formula: -HN-PEG-C(O)-X AA - wherein PEG has 1 to 50 ethylene glycol units; X AA is the amino acid sequence.

[0125] In some embodiments, the linker L2 comprises the following formula: -HN-[CH2-CH2-O] p -(CH2) 1-5 -C(O)-X AA - In the formula, p is an integer of 1 to 50, and X AA is an amino acid sequence. In some embodiments, p is an integer between 5 and 10, e.g., 5, 6, 7, 8, 9, or 10.

[0126] The amino acid portion of the linker L2 can include any suitable number of amino acid portions, as described above. For example, the amino acid sequence X AA may comprise 1 to 100 amino acid moieties, or 1 to 10 amino acid moieties, or 1 to 5 amino acid moieties.

[0127] In some embodiments, X AAis an amino acid sequence comprising 1 to 30 amino acids. AA is an amino acid sequence containing 1 to 25 amino acids, 1 to 20 amino acids, 1 to 15 amino acids, 2 to 15 amino acids, 1 to 10 amino acids, 2 to 10 amino acids, 1 to 9 amino acids, 2 to 9 amino acids, 1 to 8 amino acids, 2 to 8 amino acids, 1 to 7 amino acids, 2 to 7 amino acids, 1 to 6 amino acids, 2 to 6 amino acids, 1 to 5 amino acids, 2 to 5 amino acids, 1 to 4 amino acids, 2 to 4 amino acids, 1 to 3 amino acids, or 2 to 3 amino acids. For example, X AA can be an amino acid sequence containing 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more amino acids. AA may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid moieties. AA is an amino acid sequence comprising 1 to 10 amino acids. AA is an amino acid sequence comprising 2 to 8 amino acids. AA is an amino acid sequence comprising 2 to 6 amino acids. AA is an amino acid sequence comprising 2 to 4 amino acids. AA is an amino acid sequence comprising four amino acids. AA is an amino acid sequence comprising three amino acids. AA contains two amino acid moieties.

[0128] In some embodiments, X AA is Gly-Gly-Phe-Gly. AA is Val-Cit. AA is Ala-Ala. AA is Val-Ala.AA is Ala-Ala-Ala. AA is Val-Ala-Ala.

[0129] In some embodiments, the linker L2 comprises the following formula: -HN-PEG8-C(O)-Val-Ala- PEG8 has eight ethylene glycol units.

[0130] The linker L2 may also connect the ethylene glycol moiety to the amino acid sequence or connect the ethylene glycol or amino acid sequence to R 28 or to the compound of Formula (I) or (II). For example, the amino acid sequence may be connected to the compound of Formula (I) or (II) through a 4-aminobenzyl carboxylate group. In some embodiments, the ethylene glycol moiety is 28 In some embodiments, the linker L2 has the formula: [ka]

[0131] In some embodiments, L2 is [ka] may be selected from In the formula, X AA is the amino acid sequence, and K2 is -[CH2CH2O] 0-50 -or- [CH2] 0-12 -It is.

[0132] In some embodiments, X AA may be L-valyl-L-alanine.

[0133] R 28 R 28are azides, alkynes, bisulfones, carbohydrazides, hydrazines, hydroxylamines, iodoacetamides, isothiocyanates, maleimides, phosphines, pyridopyridazines, semihydrazides, succinimidyl esters, sulfodichlorophenol esters, sulfonyl halides, sulfosuccinimidyl esters, 4-sulfotetrafluorophenyl esters, tetrafluorophenyl esters, thiazoles, R A , O-(CH2) k -NR 26 R 26 , NHNH2, or a targeting agent, wherein the targeting agent is selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody.

[0134] Therefore, R 28 is a reactive moiety capable of reacting with a targeting agent or is the targeting agent. 28 When R is a reactive moiety, it may react with a functional group such as an aldehyde, amine, disulfide, ketone, thiol, etc. in the targeting agent, or may react with the targeting agent in a Staudinger reaction, a Pictet-Spengler reaction, and / or click chemistry. For some reactive moieties, the reactive moiety may react with the targeting agent using a suitable coupling reagent, e.g., R 28 If R is a carboxylic acid, A (CH2) j -CO2R 26 [where], a carbodiimide coupling reagent may be used.

[0135] Preferably, R 28 R is an azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semihydrazide, succinimidyl ester, and sulfonyl halides. Aor is a targeting agent, wherein the targeting agent is selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody.

[0136] In one aspect, R 28 is an azide, alkyne, bisulfone, carbohydrazide, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, semihydrazide, succinimidyl ester, and sulfonyl halide, or R A is.

[0137] Several other chemistries are known for linking compounds to antibodies. U.S. Patent No. 7,595,292 (Brocchini et al.) describes linkers that form thioesters with the sulfur in the disulfide bonds of antibodies. U.S. Patent No. 7,985,783 (Carico et al.) describes the introduction of aldehyde residues into antibodies, which are used to link compounds to antibodies.

[0138] In another embodiment, R 28 is a targeting agent, the targeting agent being selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody. The targeting agent can bind to a tumor-associated antigen, a cancer stem cell-associated antigen, or a viral antigen.

[0139] In various embodiments, the targeting agent may bind to a target selected from acute myeloid leukemia (AML M4) cells, acute promyelocytic leukemia cells, acute lymphoblastic leukemia cells, acute lymphocytic leukemia cells, chronic lymphocytic leukemia cells, chronic myeloid leukemia cells, chronic T-cell lymphocytic leukemia, myelodysplastic syndrome cells, multiple myeloma cells, prostate cancer cells, renal cell adenocarcinoma cells, pancreatic adenocarcinoma cells, lung cancer cells or gastric adenocarcinoma cells, gastric adenocarcinoma cells, breast cancer cells, colon cancer cells, melanoma cells, thyroid cancer cells, ovarian cancer cells, bladder cancer cells, liver cancer cells, head and neck cancer cells, esophageal cancer cells, Hodgkin's lymphoma cells, non-Hodgkin's lymphoma cells, mesothelioma cells, neuroblastoma cells, neuroendocrine tumor cells, neurofibromatosis type 1 (NF1) cells, neurofibromatosis type 2 (NF2) cells, or osteosarcoma cells.

[0140] In embodiments, R 28 may in a preferred embodiment be the following maleimide: [ka] It may be linked to a targeting agent.

[0141] L 2 -R 28 In an embodiment, L2-R 28 can act as a linker to connect the compound of Formula I to a targeting agent.

[0142] In some preferred embodiments, the L2-R 28 teeth, [ka] [ka] may include [ka] may include [ka] or [ka] may include It may be linked to a targeting agent.

[0143] In some other preferred embodiments, the L2-R 28 may include: [ka] [ka] It may be linked to a targeting agent.

[0144] In some preferred embodiments, the L2-R 28 teeth, [ka] [ka] may include [ka] may include [ka] may include [ka] or [ka] may include:

[0145] X 1 Preferably, X1 is O, S, NH, CH2, CH2O, C(=O), C(=O)NR 13 , N.R. 13selected from C(═O), OC(O) and C(O)—O; Preferably, X1 is O, C(=O), C(=O)NR 13 , and N.R. 13 C(=O).

[0146] More preferably, X1 is selected from O, C(=O)NH, and NHC(=O).

[0147] More preferably, X1 is O.

[0148] X 2 Preferably, X2 is O, S, NH, CH2, CH2O, C(=O), C(=O)NR 15 , N.R. 15 Selected from C(=O), OC(O), and C(O)O, or absent.

[0149] Preferably, X2 is O, C(=O), C(=O)NR 15 , and N.R. 16 C(=O) or absent.

[0150] More preferably, X2 is selected from O, C(=O)NH, and NHC(=O).

[0151] Preferably, X2 is the same as X1.

[0152] More preferably, X2 is O.

[0153] L In embodiments, L is a linker group. Preferably, any of the peptide chain, alkylene chain, paraformaldehyde chain, or polyethylene glycol chain contains one or more heteroatoms (e.g., P, N, or NH, O, and S) and / or one or more C 5-9may be interrupted or substituted by heteroarylene groups (e.g., pyrrolylene, pyrazolylene, pyrazolylene, 1,2,3-triazolylene, pyridinylene) and / or one or more phenylene groups, and each C 5-9 The heteroarylene group (e.g., pyrrolylene, pyrazolylene, pyrazolylene, 1,2,3-triazolylene, pyridinylene) and / or each phenylene group may be optionally substituted. More preferably, the chain contains from one to three heteroatoms (e.g., P, O, S, NH) and / or from one to three C 5-9 It may be interrupted by a heteroarylene group and / or one to three phenylene groups.

[0154] In some embodiments, L is an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are O, S, NH, C 5-9 which may be interrupted by or incorporated with one or more heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted. In some embodiments, peptide, alkylene, paraformaldehyde, and polyethylene glycol chains, and / or C 5-9 Heteroarylene, phenylene, heterocyclyl, and cycloalkyl moieties can be selected from one, two, or three independently selected optional R 20 Groups and / or L s -R s It may be substituted with a group.

[0155] In some embodiments, L is L2-R 28 and / or L s -R sIn some embodiments, L may include L2-R 28 In some embodiments, L may include L s -R s may include:

[0156] In some embodiments, the linker L may comprise one or more groups selected from sulfamino, sulfamyl, sulfate, sulfhydryl, sulfinamino, sulfinate, sulfino, sulfinyl, sulfinyloxy, sulfo, sulfonamido, sulfonamino, sulfonate, sulfonyl, sulfonyloxy, phosphate, phosphoramidate, thiophosphate, phosphonate, and thiophosphonate.

[0157] In some embodiments, the linker L and / or L2 comprises a moiety that allows for branching. In some embodiments, the linker L and / or L2 may comprise the following formula: [ka] In the formula, R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl, and L C is an amino acid, an amino acid derivative, a peptide chain having 2 to 6 amino acids or amino acid derivatives, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-8 -, these chains being selected from one or more P, O, S and / or NH groups, and / or C 5-9 may be interrupted by heteroarylene and / or phenylene, and each C 5-9 The heteroarylene group and / or each phenylene group may be optionally substituted.

[0158] Preferably, L is a peptide chain having 2 to 5 amino acids, 2 to 4 amino acids, or 2 to 3 amino acids, an alkylene chain containing 1 to 11 carbon atoms, 1 to 10 carbon atoms, 1 to 9 carbon atoms, 1 to 8 carbon atoms, 1 to 7 carbon atoms, 1 to 6 carbon atoms, 1 to 5 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms, which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, -(OCH2) 1-11 -, -(OCH2) 1-10 -, -(OCH2) 1-9 -, -(OCH2) 1-8 -, -(OCH2) 1-7 -, -(OCH2) 1-6 -, -(OCH2) 1-5 -, -(OCH2) 1-4 -, -(OCH2) 1-3 -, polyethylene glycol chain -(OCH2CH2) 1-5 -, chain -(OCH2CH2) 1-4 -, chain -(OCH2CH2) 1-3 -, and these chains may contain one or more heteroatoms and / or 1 to 3 C 5-9 may be interrupted by a heteroarylene group and / or 1 to 3 phenylene groups, 5-9 The heteroarylene and / or phenylene groups may be selected from one, two, or three independently selected optional R 20 Groups and / or L s -R s In some embodiments, L is optionally substituted with a L-R group. 28 and / or L s -R s one or more C optionally substituted with 5-9 In some embodiments, L includes heteroarylene and / or phenylene groups. 28 one or more C optionally substituted with 5-9 In some embodiments, L includes heteroarylene and / or phenylene groups. s -R s one or more C optionally substituted with 5-9Contains heteroarylene and / or phenylene groups.

[0159] More preferably, L may be selected from alkylene chains containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds.

[0160] More preferably, L may be selected from CH=CH, CH2, CH2CH2, CH2CH2CH2, CH2CH2CH2CH2, and CH2CH2CH2CH2CH2CH2.

[0161] G In some embodiments, the D moiety of formula (I) comprises a G that is a G-alkylated DNA group of formula (II): [ka] During the ceremony, The dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4; The wavy line indicates the point of attachment to Q; m is 0 or 1, R1, R3, and R4 are independently H, -L S -R S , and R 29 and R2 is selected from H, L2-R 28 , R 29 , and -L S -R S or one of R1 and R2, R2 and R3, or R3 and R4, taken together with the carbon atom to which they are attached, is selected from a 6-membered aryl, or one, two, or three independently selected, optionally -L S -R S and R 20 forming a 5- or 6-membered cyclic, heterocyclic, or heteroaryl ring optionally substituted with a group, R5 and R6 are: (i) R5 is H, OH, -L S -R S and O.C. 1-6alkyl, and R6 is selected from H, SO3H, -L S -R S , nitrogen protecting group, -L2-R 28 , and R A or (ii) R5 is oxo or H and R6 is H or C 1-6 alkyl, or (iii) R5 and R6 are selected so that they together form a double bond; R7 and R9 are independently H, -L S -R S , and R 20 is selected from R8 is H, SR 24 , SCH2Ph, R 20 , L2-R 28 , and -L S -R S is selected from R A is (CH2) j -OH, (CH2) j -CO2R 26 , C(=O)-O-(CH2) k -NR 26 R 27 , (CH2) j NR 26 R 27 , C(=O)-NH-(CH2) j -NR 26 R 27 and C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 is selected from L2 is a bond or a linker moiety having 1 to 200 non-hydrogen atoms selected from C, N, O, S, or halogen, and optionally incorporating an ether, oxo, carboxamidyl, uretanyl, branched, cyclic, unsaturated, heterocyclyl, aryl, or heteroaryl moiety; R 28are azides, alkynes, bisulfones, carbohydrazides, hydrazines, hydroxylamines, iodoacetamides, isothiocyanates, maleimides, phosphines, pyridopyridazines, semihydrazides, succinimidyl esters, sulfodichlorophenol esters, sulfonyl halides, sulfosuccinimidyl esters, 4-sulfotetrafluorophenyl esters, tetrafluorophenyl esters, thiazoles, R A , O-(CH2) k -NR 26 R 26 , NHNH2, or a targeting agent selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; Each R 29 are independently, R 20 , R 21 , =CH2, =CH-(CH2) s -CH3, =CH-(CH2) s -R 21 , =O, (CH2) s -OR 21 , (CH2) s -CO2R 21 , (CH2) s -NR 21 R 24 , O-(CH2) t -NR 21 R 24 , NH-C(O)-R 21 , O-(CH2) t -NH-C(O)-R 21 , O-(CH2) t -C(O)-NH-R 21 , (CH2) s -SO2R 21 , O-SO2R 21 , (CH2) s -C(O)R 21 and (CH2) s -C(O)NR 21 R 24 is selected from Each R 20are independently F, Cl, Br, (CH2) j -OH, C 1-6 Alkyl, OC 1-6 Alkyl, OCH2Ph, (CH2) j -CO2R 26 , O-(CH2) k -NR 26 R 27 , C(=O)-O-(CH2) k -NR 26 R 27 , C(=O)-NR 26 R 27 , (CH2) j -NR 26 R 27 , N.R. 26 NH2, C(=O)-NH-(CH2) j -NR 26 R 27 , C(=O)-NH-C6H4-(CH2) j -R 26 , C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 , -L2-R 28 , S(O)2-(C 1-6 alkyl), O-(CH2) k -O-(C 1-6 alkyl), (CH2) j -S(O)2-NR 26 R 27 , C(=NH)-O-(C 1-6 alkyl), (CH2) k -O-(C 1-6 alkyl), CN, NCO, Cy, C(O)-NH-(CH2) j -Cy, C(O)-Cy, NH-C(O)-NR 26 R 27 is selected from [ka] each j and s is independently selected from 0, 1, 2, 3, 4, 5, or 6; each k and t is independently selected from 1, 2, 3, 4, 5, or 6; Each R 21 are independently H, C1-12 Alkyl, C 5-6 Heterocyclyl, C 5-9 Heteroaryl, C 6-15 Heteroarylalkyl, phenyl, and C 7-12 and aralkyl groups, wherein the heterocyclyl, heteroaryl, heteroarylalkyl, phenyl, and aralkyl groups are selected from one, two, or three independently selected optional R 20 may be substituted with a group, Each R 24 , R 26 , and R 27 are independently H and C 1-12 alkyl, Each Cy independently represents C 5-6 Heterocyclyl or C 5-6 heteroaryl groups, wherein the heterocyclyl or heteroaryl group is selected from one or two R 20 may be substituted with a group, L S is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are O, S, NH, C 5-9 which may be interrupted by or incorporate one or more heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; C 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L S teeth, [ka] and R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0162] In some embodiments, L S teeth, [ka] where: L C is an amino acid, an amino acid derivative, a peptide chain having 2 to 6 amino acids or amino acid derivatives, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-8 -, and these chains contain one or more groups selected from one or more P, O, S and / or NH groups, and / or C 5-9 may be interrupted by heteroarylene and / or phenylene, and each C 5-9 the heteroarylene group and / or each phenylene group may be optionally substituted; R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0163] In some embodiments, in the G moiety of formula (II), at least one of R, R, R, R, R, R, R, R, R, and R is -L S -R S In some embodiments, in the G moiety of formula (II), R is -L S -R S In some embodiments, in the G moiety of formula (II), R2 is R S In some embodiments, in the G moiety of formula (II), R is -L S -R S In some embodiments, in the G moiety of formula (II), R2 is R S In some embodiments, in the G moiety of formula (II), R3 is -L S -R S In some embodiments, in the G moiety of formula (II), R3 is R SIn some embodiments, in the G moiety of formula (II), R4 is -L S -R S In some embodiments, in the G moiety of formula (II), R4 is R S In some embodiments, in the G moiety of formula (II), R5 is -L S -R S In some embodiments, in the G moiety of formula (II), R5 is R S In some embodiments, in the G moiety of formula (II), R6 is -L S -R S In some embodiments, in the G moiety of formula (II), R6 is R S In some embodiments, in the G moiety of formula (II), R7 is -L S -R S In some embodiments, in the G moiety of formula (II), R7 is R S In some embodiments, in the G moiety of formula (II), R8 is -L S -R S In some embodiments, in the G moiety of formula (II), R is R S In some embodiments, in the G moiety of formula (II), R9 is -L S -R S In some embodiments, in the G moiety of formula (II), R is R S is.

[0164] In some embodiments, the G-alkylating agent unit G comprises a group of formula (II): [ka] During the ceremony, The dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4; The wavy line indicates the point of attachment to Q; m is 0 or 1, R1, R3, and R4 are independently selected from H and R 29 and R2 is selected from H, L2-R 28、 R29 , and R S or one of R1 and R2, R2 and R3, or R3 and R4, together with the carbon atom to which they are attached, is selected from a 6-membered aryl, or one, two, or three independently selected optional R 20 forming a 5- or 6-membered cyclic, heterocyclic or heteroaryl ring optionally substituted by a group, R5 and R6 are: (i) R5 is H, OH, and OC 1-6 alkyl, and R6 is selected from H, SO3H, -L S -R S , nitrogen protecting group, -L2-R 28 and R A or (ii) R5 is oxo or H and R6 is H or C 1-6 alkyl, or (iii) R5 and R6 together form a double bond; R7 and R9 are independently H and R 20 is selected from R8 is H, SR 24 , SCH2Ph, R 20 , L2-R 28 , and R S is selected from R A is (CH2) j -OH, (CH2) j -CO2R 26 , C(=O)-O-(CH2) k -NR 26 R 27 , (CH2) j NR 26 R 27 , C(=O)-NH-(CH2) j -NR 26 R 27 and C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 is selected from L2 is a bond or a linker moiety having 1 to 200 non-hydrogen atoms selected from C, N, O, S, or halogen, and optionally incorporating an ether, oxo, carboxamidyl, uretanyl, branched, cyclic, unsaturated, heterocyclyl, aryl, or heteroaryl moiety; R 28 are azides, alkynes, bisulfones, carbohydrazides, hydrazines, hydroxylamines, iodoacetamides, isothiocyanates, maleimides, phosphines, pyridopyridazines, semihydrazides, succinimidyl esters, sulfodichlorophenol esters, sulfonyl halides, sulfosuccinimidyl esters, 4-sulfotetrafluorophenyl esters, tetrafluorophenyl esters, thiazoles, R A , O-(CH2) k -NR 26 R 26 , NHNH2, or a targeting agent selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; Each R 29 are independently, R 20 , R 21 , =CH2, =CH-(CH2) s -CH3, =CH-(CH2) s -R 21 , =O, (CH2) s -OR 21 , (CH2) s -CO2R 21 , (CH2) s -NR 21 R 24 , O-(CH2) t -NR 21 R 24 , NH-C(O)-R 21 , O-(CH2) t -NH-C(O)-R 21 , O-(CH2) t -C(O)-NH-R 21 , (CH2) s -SO2R 21, O-SO2R 21 , (CH2) s -C(O)R 21 and (CH2) s -C(O)NR 21 R 24 is selected from Each R 20 are independently F, Cl, Br, (CH2) j -OH, C 1-6 Alkyl, OC 1-6 Alkyl, OCH2Ph, (CH2) j -CO2R 26 , O-(CH2) k -NR 26 R 27 , C(=O)-O-(CH2) k -NR 26 R 27 , C(=O)-NR 26 R 27 , (CH2) j -NR 26 R 27 , N.R. 26 NH2, C(=O)-NH-(CH2) j -NR 26 R 27 , C(=O)-NH-C6H4-(CH2) j -R 26 , C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 , -L2-R 28 , S(O)2-(C 1-6 alkyl), O-(CH2) k -O-(C 1-6 alkyl), (CH2) j -S(O)2-NR 26 R 27 , C(=NH)-O-(C 1-6 alkyl), (CH2) k -O-(C 1-6 alkyl), CN, NCO, Cy, C(O)-NH-(CH2) j -Cy, C(O)-Cy, NH-C(O)-NR 26 R 27 is selected from [ka] each j and s is independently selected from 0, 1, 2, 3, 4, 5, or 6; each k and t is independently selected from 1, 2, 3, 4, 5, or 6; Each R 21 are independently H, C 1-12 Alkyl, C 5-6 Heterocyclyl, C 5-9 Heteroaryl, C 6-15 Heteroarylalkyl, phenyl, and C 7-12 and aralkyl groups, wherein the heterocyclyl, heteroaryl, heteroarylalkyl, phenyl, and aralkyl groups are selected from one, two, or three independently selected optional R 20 may be substituted with a group, Each R 24 , R 26 , and R 27 are independently H and C 1-12 alkyl, Each Cy independently represents C 5-6 Heterocyclyl or C 5-6 heteroaryl groups, wherein the heterocyclyl or heteroaryl group is selected from one or two R 20 may be substituted with a group, L S is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are O, S, NH, C 5-9 which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L S teeth, [ka] and R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0165] In some embodiments, in the G moiety of formula (II), at least one of R2, R6, and R8 is -L S -R S In some embodiments, in the G moiety of formula (II), R2 is -L S -R S In some embodiments, in the G moiety of formula (II), R2 is R S In some embodiments, in the G moiety of formula (II), R6 is -L S -R S In some embodiments, in the G moiety of formula (II), R6 is R S In some embodiments, in the G moiety of formula (II), R8 is -L S -R S In some embodiments, in the G moiety of formula (II), R is R S is.

[0166] In some embodiments, the G-alkylating agent unit G comprises a group of formula (II-A): [ka] wherein the substituents have the definitions given above.

[0167] In some embodiments, G is selected from the group G1-G8 below. [ka] [ka]

[0168] In some embodiments, G is selected from the group G1-A to G8-A below. [ka] [ka]

[0169] In some embodiments, the compound according to formula (I) can be a compound according to formula (IV): [ka]

[0170] In some embodiments, in formula (IV), at least one of R2, R5, R6, and R8 is -L S -R S In some embodiments, in formula (IV), R2 is -L S -R S In some embodiments, in formula (IV), R2 is R S In some embodiments, in formula (IV), R5 is -L S -R S In some embodiments, in formula (IV), R5 is R S In some embodiments, in formula (IV), R6 is -L S -R S In some embodiments, in formula (IV), R6 is R S In some embodiments, in formula (IV), R8 is -L S -R S In some embodiments, in formula (IV), R is R S is.

[0171] In some embodiments, the compound according to formula (I) can be a compound according to formula (IV-A): [ka] wherein the substituents have the definitions given above.

[0172] In some embodiments, the compound according to formula (I) can be a compound selected from the group consisting of: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] In the formula, R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0173] In some embodiments, the compound according to formula (I) can be a compound selected from the group consisting of: [ka] [ka] [ka] In the formula, R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0174] A In some embodiments, the D moiety of formula (I) comprises an A-alkylated DNA group. As used herein, unless the context requires otherwise, the seco and spiro forms of an A group are used interchangeably. Thus, the A group of a seco form of an A group includes the spiro form of an A group, and the spiro form of an A group includes the seco form of an A group.

[0175] In some embodiments, A is an A-alkylated DNA group of formula (IIIa) or (IIIb): [ka] During the ceremony, Z1 is a leaving group, optionally a halide, triflate, or tosylate; X is CR 17 , N, NR 17 , S or O, and the dashed line to X indicates the optional presence of a double bond depending on the nature of X, R 17 is H, -L S -R S , or R 20 and z is 0 or 1, '''' is OH or -L S -R S and L S is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are O, S, NH, C 5-9 which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L S teeth, [ka] and R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0176] In some embodiments, the A-alkylated DNA group of formula (IIIa) or (IIIb) comprises at least one -L S -R S In some embodiments, in the A moiety of formula (IIIa), X comprises a CR 17 and R 17 -L S -R S In some embodiments, in the A moiety of formula (IIIa), X is CR 17 and R 17 is R S In some embodiments, in the A moiety of formula (IIIa), X is NR 17 and R 17 -L S -R S In some embodiments, in the A moiety of formula (IIIa), X is NR 17 and R 17 is R S In some embodiments, in the A moiety of formula (IIIa), "" is -L S -R S In some embodiments, in the A moiety of formula (IIIa), "" is R S In some embodiments, in the A moiety of formula (IIIb), X is CR 17 and R 17 -L S -R S In some embodiments, in the A moiety of formula (IIIb), X is CR 17 and R 17 is R S In some embodiments, in the A moiety of formula (IIIb), X is NR 17 and R 17 -L S -RS In some embodiments, in the A moiety of formula (IIIb), X is NR 17 and R 17 is R S is.

[0177] In some embodiments, the compound according to formula (I) can be a compound selected from the group consisting of: [ka] [ka] [ka] [ka] In the formula, R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0178] In some embodiments, the compound according to formula (I) can be a compound selected from the group consisting of: [ka] [ka] In the formula, R S is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

[0179] For purposes herein, a "receptor human framework" is a framework comprising the amino acid sequence of a light chain variable domain (VL) framework or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human consensus framework, as defined below. A receptor human framework "derived from" a human immunoglobulin framework or a human consensus framework may comprise the same amino acid sequence or may contain amino acid sequence changes. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the VL receptor human framework is identical in sequence to the VL human immunoglobulin framework sequence or the human consensus framework sequence.

[0180] "Affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by conventional methods known in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

[0181] An "affinity matured" antibody refers to an antibody with one or more alterations in one or more hypervariable regions (HVRs), compared to a parent antibody that does not possess such alterations, which alterations result in an improvement in the affinity of the antibody for antigen.

[0182] The term "antibody" herein is used in the broadest sense and encompasses a variety of antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as the antibody exhibits the desired antigen-binding activity.

[0183] The term "antibody fragment" refers to a molecule other than an intact antibody that contains a portion of the intact antibody and that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fa", Fa"-SH, F(a")2, diabodies, linear antibodies, single-chain antibody molecules (e.g., scFv), and multispecific antibodies formed from antibody fragments.

[0184] The term "chimeric" antibody refers to an antibody in which a portion of the heavy and / or light chain is derived from one particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.

[0185] The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into subclasses (isotypes), e.g., IgGi, IgG2, IgG3, IgG4, IgAi, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0186] As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents the function of cells and / or causes the death or destruction of cells. Cytotoxic agents include radioisotopes (e.g., At 211 , I 131 , I 125 , Y 90 ,Re 186 , Re188 , Sm 153 , Bi 212 , P 32 , Pb 212and radioactive isotopes of Lu), chemotherapeutic agents or drugs (e.g., methotrexate, adriamicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents), growth inhibitory agents, enzymes such as nucleolytic enzymes and fragments thereof, antibiotics, toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and / or variants thereof, and various anti-tumor or anti-cancer agents disclosed below.

[0187] "Co-administration" means administering two (or more) drugs intravenously during the same administration, rather than sequential infusion of two or more drugs. Generally, this may involve combining two (or more) drugs together in the same IV bag before administering them simultaneously.

[0188] A drug that is administered "concurrently" with one or more other drugs is administered during the same treatment cycle, on the same treatment day as the one or more other drugs, and optionally, simultaneously with the one or more other drugs. For example, in a cancer therapy administered every three weeks, the concurrently administered drugs are each administered on day 1 of a three-week cycle.

[0189] "Chemotherapeutic agent" refers to a compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (CYTOXAN®); alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylomelanin. ethylenimines and methylamelamines, including acetogenins (especially bullatacin and bullatacinone); delta-9-tetrahydrocannabinol (dronabinol, MARINOL®); beta-lapachone; lapachol; colchicine; betulinic acid; camptothecin (synthetic analogs topotecan (HYCAMTIN®), CPT-11 (irinotecan, CAMPTOSAR®)), including acetylcamptothecin, scopolectin, and 9-aminocamptothecin; bryostatin; callystatin; CC-1065 (including its synthetic analogs adozelesin, carzelesin, and bizelesin); podophyllotoxin; podophyllic acid; teniposide; cryptophycins (especially cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycins (synthetic analogs, KW-2 189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, fenesterine, prednimustine, trofosfamide, nitrogen mustards such as uracil mustard;nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; enediyne antibiotics (e.g., antibiotics such as calicheamicin, particularly calicheamicin gamma 11 and calicheamicin omega 11 (see, e.g., Nicolaou et al., Angew. Chem Intl. Ed. Engl., 33:183-186 (1994)); oral alpha-4 integrin inhibitors, CDP323; dynemicins, including dynemicin A; esperamicin; and neocarzinostatin chromophores and related chromoprotein enediyne antibiotic chromophores), aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, carabicin, carminomycin (carminomycin), carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (ADRIAMYCIN®), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, doxorubicin hydrochloride liposomal injection (DOXIL®), liposomal doxorubicin TLC D-99 (MYOCET®), pegylated liposomal doxorubicin (including CAELYX® and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin;Antimetabolites such as methotrexate, gemcitabine (GEMZAR®), tegafur (UFTORAL®), capecitabine (XELODA®), epothilones, and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; acitabine Pyrimidine analogues such as olubilizede, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine; androgens such as calucelone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; anti-adrenal drugs such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as furoic acid; aceglatone; and aldophosphamide glycosides. glycoside); aminolevulinic acid; eniluracil; amsacrine; bestravcil; bisantrene; XXX olubilizee; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; epothilone; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, Oregon); razoxane; rhizoxin; schizophyllan; spirogermanium; tenuazonic acid; triazicon;2,2',2'-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A, and anguidine); urethanes; vindesine (ELDISINE®, FILDESIN®); dacarbazine; mannommustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinosides ("Ara-"); thiotepa; taxoids such as paclitaxel (TAXOL®), albumin-engineered nanoparticle formulations of paclitaxel (ABRAXANETM), and docetaxel (TAXOTERE®); XXX orvirized XXX1; 6-thioguanine; mercaptopurine; methotrexate; cisplatin platinum agents such as oxaliplatin (e.g., ELOXATIN®) and carboplatin; vincas that prevent tubulin polymerization to form microtubules, including vinblastine (VELBAN®), vincristine (ONCOVIN®), vindesine (ELDISINE®, FILDESIN®), and vinorelbine (NAVELBINE®); etoposide (VP-16); ifosfamide; mitoxantrone; leucovorin; novantrone; edatrexate; daunomycin; aminopterin; ibandronate; the topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid, including bexarotene (TARGRETIN®);bisphosphonates such as clodronate (e.g., BONEFOS® or OSTAC®), etidronate (DIDROCAL®), NE-58095, zoledronic acid / zoledronate (ZOMETA®), alendronate (FOSAMAX®), pamidronate (AREDIA®), tiludronate (SKELID®), or risedronate (ACTONEL®); troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); antisense oligonucleotides, which inhibit the expression of genes, particularly in signal transduction pathways involved in abnormal cell proliferation, such as PKC-alpha, Raf, H-Ras, and epidermal growth factor receptor (EGF-R); vaccines, such as the THERATOPE® vaccine and gene therapy vaccines, such as the ALLOVECTIN® vaccine, LEUVECTIN® vaccine, vaccines such as the VAXID® vaccine and the topoisomerase 1 inhibitors (e.g., LURTOTECAN®); rmRH (e.g., ABARELIX®); BAY439006 (sorafenib, Bayer); SU-11248 (sunitinib, SUTENT®, Pfizer); perifosine, COX-2 inhibitors (e.g., celecoxib or etoricoxib), proteosome inhibitors (e.g., PS 341); bortezomib (VELCADE®); CCI-779; tipifarnib (R11577); orafenib, ABT510; Bcl-2 inhibitors such as oblimersen sodium (GENASENSE®); pixantrone; EGFR inhibitors; tyrosine kinase inhibitors; serine-threonine kinase inhibitors such as rapamycin (sirolimus, RAPAMUNE®); farnesyltransferase inhibitors such as lonafarnib (SCH 6636, SARASAR trademark);and pharmaceutically acceptable salts, acids, or derivatives of any of the above, as well as combinations of two or more of the above, such as CHOP, an abbreviation for the combination therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone, and FOLFOX, an abbreviation for the treatment of oxaliplatin (ELOXATIN trademark) in combination with 5-FU and leucovorin;

[0190] Chemotherapeutic agents, as defined herein, include "antihormonal agents" or "endocrine therapeutic agents," which act to regulate, reduce, block, or inhibit the effects of hormones that can promote cancer growth. These can be hormones themselves, antiestrogens with mixed agonist / antagonist profiles, including tamoxifen (NOLVADEX®), 4-hydroxytamoxifen, toremifene (FARESTON®), idoxifene, droloxifene, raloxifene (EVISTA®), trioxifene, keoxifene, and selective estrogen receptor modulators (SERMs) such as SERM3; agonist profiles such as fulvestrant (FASLODEX®) and EM800 (such agents can block estrogen receptor (ER) dimerization, inhibit DNA binding, increase ER turnover, and / or suppress ER levels). aromatase inhibitors, including steroidal aromatase inhibitors such as formestane and exemestane (AROMASIN®) and nonsteroidal aromatase inhibitors such as anastrozole (ARFMIDEX®), letrozole (FEMARA®), and aminoglutethimide, and other aromatase inhibitors, including vorozole (RIVISOR®), megestrol acetate (MEGASE®), fadrozole, and 4(5)-imidazole; luteinizing hormone-releasing hormone agonists, including leuprolide (LUPRON® and ELIGARD®), goserelin, buserelin, and tripterelin;Sex steroids, including progestins such as megestrol acetate and medroxyprogesterone acetate, estrogens such as diethylstilbestrol and olubilin, and androgens / retinoids such as fluoxymesterone, all-trans-retinoic acid, and fenretinide; onapristone; antiprogesterones; estrogen receptor downregulators (ERDs); antiandrogens such as flutamide, nilutamide, and bicalutamide, as well as pharmaceutically acceptable salts, acids, or derivatives of any of the above, and combinations of two or more of the above.

[0191] "Drug," "drug substance," "active pharmaceutical ingredient," and the like refer to compounds (e.g., compounds of Formula (I), compounds of Formula (IV), and other formulae described herein, as well as compounds specifically named above) that can be used to treat a subject in need thereof.

[0192] "Conjugate" refers to a compound or construct comprising (a) at least one of the Formula (I), Formula (IV) drug compounds and compounds of other formulae described herein, as well as compounds specifically named above, and (b) at least one targeting agent (e.g., a protein, portion of a protein, peptide, nucleic acid, antibody, antibody fragment, hormone, etc.).

[0193] "Effector function" refers to the biological activity attributable to the Fc region of an antibody, which varies depending on the antibody isotype. Examples of antibody effector functions include Clq binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0194] The term "epitope" refers to the specific site on an antigen molecule to which an antibody binds.

[0195] "Epitope 4D5" or "4D5 epitope" or "4D5" refers to the region within the extracellular domain of HER2 to which antibody 4D5 (ATCC CRL 10463) and trastuzumab bind. This epitope is located near the transmembrane domain of HER2, within domain IV of HER2. To screen for antibodies that bind to the 4D5 epitope, a conventional cross-blocking assay, such as that described in "Antibodies, A Laboratory Manual," Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988), can be performed. Alternatively, epitope mapping can be performed to assess whether an antibody binds to the 4D5 epitope of HER2 (e.g., any one or more residues encompassed within the region from about residue 550 to about residue 610 of HER2 (SEQ ID NO: 39)).

[0196] "Epitope 2C4" or "2C4 epitope" refers to the region in the extracellular domain of HER2 that antibody 2C4 binds to. To screen for antibodies that bind to the 2C4 epitope, conventional cross-blocking assays such as those described in "Antibodies, A Laboratory Manual," Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988) can be performed. Alternatively, epitope mapping can be performed to evaluate whether an antibody binds to the 2C4 epitope of HER2. Epitope 2C4 comprises residues from domain II in the extracellular domain of HER2. 2C4 antibody and pertuzumab bind to the extracellular domain of HER2 at the junction of domains I, II, and III (Franklin et al. Cancer Cell 5:317-328 (2004)). Anti-HER2 mouse antibody 7C2 binds to an epitope in domain I of HER2. For example, see PCT application publication WO 98 / 17797. This epitope is different from the epitope bound by trastuzumab, which binds to domain IV of HER2, and the epitope bound by pertuzumab, which binds to domain II of HER2. By binding to domain IV, trastuzumab disrupts the ligand-independent HER2-HER3 complex, thereby inhibiting downstream signal transduction (e.g., PI3K / AKT). In contrast, the binding of pertuzumab to domain II prevents the ligand-driven interaction of HER2 with other HER family members (e.g., HER3, HER1, or HER4), and therefore also prevents downstream signal transduction. The binding of mAb 7C2 to domain I does not interfere with the binding of trastuzumab or pertuzumab to domains IV and II, respectively, thereby offering the possibility of combining ADCs of mAb 7C2 with trastuzumab, trastuzumab emtansine (T-DM-1), and / or pertuzumab. Murine antibodies 7C2 and 7C2.B9 are described in PCT Publication WO 98 / 17797.The anti-HER2 7C2 humanized antibody is disclosed in WO 2016 / 040723 A1.

[0197] "Excipient" refers to any substance that can affect the bioavailability of a drug but is pharmacologically inactive.

[0198] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain containing at least a portion of the constant region. This term includes native sequence Fc regions and variant Fc regions. In one embodiment, a human IgG heavy chain Fc region extends from Cys226, or from Pro230, to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is in accordance with Kabat et al., Sequences of Proteins of Immunological Interest, 5t h According to the EU numbering system, also known as the EU index, as described in Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0199] "Framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences generally appear in the following order within VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0200] The terms "full length antibody," "intact antibody," and "whole antibody" are used interchangeably herein and refer to an antibody having a structure substantially similar to a native antibody structure or an antibody having a heavy chain containing an Fc region as defined herein.

[0201] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," including the primary transformed cell and progeny derived therefrom, regardless of the number of passages. The progeny may not be completely identical in nucleic acid content to the parent cell, but may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.

[0202] A "human antibody" is an antibody having an amino acid sequence that corresponds to that of an antibody produced by a human or human cell, or derived from a non-human source that utilizes the human antibody repertoire or other human antibody coding sequences. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues.

[0203] A "human consensus framework" is a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. Generally, the subgroup of sequences is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda, MD (1991), vols. 1-3. In one embodiment, for VL, the subgroup is subgroup kappa I as in Kabat et al., supra. In one embodiment, for VH, the subgroup is subgroup III as in Kabat et al., supra.

[0204] A "humanized" antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the HVRs (e.g., "complementarity-determining regions," CDRs) correspond to HVRs of a non-human antibody and all or substantially all of the FRs correspond to FRs of a human antibody. A humanized antibody may optionally comprise at least a portion of an antibody constant region derived from a human antibody. A "humanized form" of an antibody, e.g., a non-human antibody, refers to an antibody that has undergone humanization.

[0205] As used herein, the term "hypervariable region" or "HVR" refers to each of the regions of an antibody variable domain that are hypervariable in sequence and / or form structurally defined loops ("hypervariable loops"). Typically, naturally occurring four-chain antibodies contain six HVRs, three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). HVRs generally contain amino acid residues from the hypervariable loops and / or CDRs that have the highest sequence variability and / or are involved in antigen recognition. Exemplary hypervariable loops occur at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3). (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987).) Exemplary CDRs (CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3) occur at amino acid residues 24-34 of L1, 50-56 of L2, 89-97 of L3, 31-35B of H1, 50-65 of H2, and 95-102 of H3. (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991). With the exception of CDR1 in VH, CDRs generally contain amino acid residues that form hypervariable loops. CDRs also contain "specificity-determining residues" or "SDRs," which are residues that contact the antigen. SDRs are contained within regions of CDRs called truncated CDRs or a-CDRs. Exemplary a-CDRs (a-CDR-L1, a-CDR-L2, a-CDR-L3, a-CDR-H1, a-CDR-H2, and a-CDR-H3) occur at amino acid residues 31-34 of L1, 50-55 of L2, 89-96 of L3, 31-35B of H1, 50-58 of H2, and 95-102 of H3. (See Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)). Unless otherwise indicated, HVR residues and other residues in the variable domain (e.g., FR residues) are numbered herein according to Kabat et al., supra.

[0206] An "immunoconjugate" is an antibody conjugated to one or more heterologous molecules, including, but not limited to, cytotoxic agents.

[0207] As used herein with respect to adjunctive therapy, the term "immunosuppressant" refers to a substance that acts to suppress or mask the immune system of the mammal being treated herein. This would include substances that suppress cytokine production, downregulate or suppress autoantigen expression, or mask MHC antigens. Examples of such agents include 2-amino-6-aryl-5-substituted pyrimidines (see U.S. Pat. No. 4,65,077); nonsteroidal anti-inflammatory drugs (NSAIDs); glucocorticoids such as ganciclovir, tacrolimus, cortisol, or aldosterone, anti-inflammatory drugs such as cyclooxygenase inhibitors, 5-lipoxygenase inhibitors, or leukotriene receptor antagonists; purine antagonists such as azathioprine or mycophenolate mofetil (MMF); alkylating agents such as cyclophosphamide; XXX orvirizade XXXie; danazol; dapsone; glutaraldehyde (which masks MHC antigens, as described in U.S. Pat. No. 4,120,649); anti-idiotypic antibodies against MHC antigens and MHC fragments; cyclosporin A; steroids such as corticosteroids or glucocorticoids or glucocorticoid analogs, e.g., prednisone, sol methylprednisolone, including SOLU-MEDROL® methylprednisolone sodium succinate, and dexamethasone; dihydrofolate reductase inhibitors such as methotrexate (oral or subcutaneous); antimalarials such as chloroquine and hydroxychloroquine; sulfasalazine; leflunomide; cytokine or cytokine receptor antibodies, including anti-interferon-alpha, -beta, or -gamma antibodies, anti-tumor necrosis factor (TNF)-alpha antibodies (infliximab (REMICADE® or adalimumab), anti-TNF-alpha immunoadhesin (etanercept), anti-TNF-beta antibodies, anti-interleukin-2 (IL-2) antibodies and anti-IL-2 receptor antibodies, and anti-interleukin-6 (IL-6) receptor antibodies and antagonists (e.g., ACTEMRA® (tocilizumab));anti-LFA-1 antibodies, including anti-CD11a and anti-CD18 antibodies; anti-L3T4 antibodies; xenogeneic antilymphocyte globulins; pan-T antibodies, preferably anti-CD3 or anti-CD4 / CD4a antibodies; soluble peptides containing an LFA-3 binding domain (WO 90 / 08187); streptokinase; transforming growth factor-beta (TGF-beta); streptodornase; host-derived RNA or DNA; FK506; RS-61443; chlorambucil; deoxyspergualin; rapamycin; T cell receptors (Cohen et al., U.S. Pat. No. 5,114,721); T cell receptor fragments (Offner et al. al, Science, 251:430-432 (1991), WO 90 / 11294, Ianeway, Nature, 341:482 (1989), and WO 91 / 01133; BAFF antagonists and zTNF4 antagonists, such as BAFF antibodies and BR3 antibodies (for review, see also Mackay and Mackay, Trends Immunol, 23:113-5 (2002), and the definitions below); CD40-CD40 ligand (e.g., Durie et al, Science, 261:1328-30 (1993); Mohan et al, J. Immunol, 154:1470-80 (1995)) and CTLA4-Ig (Finck et al, Biologic agents that interfere with T cell helper signals, such as anti-CD40 receptor or anti-CD40 ligand (CD154), including blocking antibodies against CD40 receptor (e.g., Br. et al., Science, 265:1225-7 (1994)); and T cell receptor antibodies such as T10B9 (European Patent No. 340,109). Some preferred immunosuppressants herein include cyclophosphamide, chlorambucil, azathioprine, leflunomide, MMF, or methotrexate.

[0208] An "isolated antibody" is an antibody that has been separated from a component of its natural environment. In some embodiments, the antibody is purified to greater than 95% or 99% purity, as determined, for example, by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse-phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chromatogr. B 848:79-87 (2007).

[0209] "Isolated nucleic acid" refers to a nucleic acid molecule that has been separated from a component of its natural environment. Isolated nucleic acid includes a nucleic acid molecule contained in cells that ordinarily contain the nucleic acid molecule, but where the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0210] An "isolated nucleic acid encoding an antibody" refers to one or more nucleic acid molecules encoding the antibody heavy and light chains (or fragments thereof), including such nucleic acid molecules in a single vector or separate vectors, and including such nucleic acid molecules present in one or more locations within a host cell.

[0211] As used herein, the term "human epidermal growth factor receptor 2" (HER2) refers to any naturally occurring mature HER2 resulting from processing of the HER2 precursor protein in cells. The term is generally taken to refer to a protein involved in normal cell growth. Human epidermal growth factor receptor 2 can be produced in higher than normal amounts by some types of cancer cells, including breast, ovarian, bladder, pancreatic, and gastric cancers. This can cause the cancer cells to grow more rapidly and metastasize to other parts of the body. As used herein, the term includes HER2 from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus monkeys) and rodents (e.g., mice and rats), unless otherwise indicated. The term also includes naturally occurring variants of HER2, such as splice variants or allelic variants. The amino acid sequence of an exemplary human HER2 precursor protein with a signal sequence (having amino acids 1-22) is set forth in SEQ ID NO: 64. An exemplary amino acid sequence of mature human HER2 is amino acids 23 to 1255 of SEQ ID NO:64.

[0212] The term "HER2-positive cells" refers to cells that express HER2 on their surface. As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies comprising the population are identical and / or bind to the same epitope, but excludes variant antibodies that contain, for example, natural mutations or that may arise during the production of the monoclonal antibody preparation, and such variants are generally present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present disclosure may be made by a variety of techniques, including, but not limited to, hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of the human immunoglobulin loci; such methods and other exemplary methods for making monoclonal antibodies are described herein.

[0213] A "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (e.g., a cytotoxic moiety) or a radiolabel. Naked antibodies may be present in pharmaceutical formulations.

[0214] "Native antibody" refers to a naturally occurring immunoglobulin molecule with a variety of structures. For example, a natural IgG antibody is a heterotetrameric glycoprotein of approximately 150,000 daltons, composed of two identical light chains and two identical heavy chains disulfide-bonded. Each heavy chain contains, from N- to C-terminus, a variable region (VH), also called a variable heavy domain or heavy chain variable domain, followed by three constant domains (CHI, CH2, and CH3). Similarly, each light chain contains, from N- to C-terminus, a variable region (VL), also called a variable light domain or light chain variable domain, followed by a constant light (CL) domain. Based on the amino acid sequence of their constant domains, the light chains of an antibody can be assigned to one of two types, called kappa (κ) and lambda (λ).

[0215] "Percent (%) amino acid sequence identity" to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to those in the reference polypeptide sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in a variety of ways within the skill of the art, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MegAlign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared. However, for purposes of this specification, percent amino acid sequence identity values ​​are generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program is the copyright of Genentech, Inc., and its source code has been submitted to the U.S. Copyright Office, Washington, DC 20559, USA, as user documentation, and is registered under U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California, or can be compiled from its source code. The ALIGN-2 program must be compiled for use on UNIX operating systems, including Digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary. In situations where ALIGN-2 is used for amino acid sequence comparison, the percent amino acid sequence identity of a given amino acid sequence A to a given amino acid sequence B (alternatively, it can be referred to as a given amino acid sequence A having or containing a particular percent amino acid sequence identity to a given amino acid sequence B, with a given amino acid sequence B, or to a given amino acid sequence B) is calculated as follows: 100×X / Y ratio where X is the number of amino acid residues scored as an identity match by the sequence alignment program ALIGN-2 in that program's alignment of A and B, and where Y is the total number of amino acid residues in B. It is understood that if the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not be equal to the % amino acid sequence identity of B to A. Unless specifically stated otherwise, all values ​​of % amino acid sequence identity used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[0216] The term "PD-1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-1 axis binding partner with one or more of its binding partners to eliminate T cell dysfunction resulting from signaling along the PD-1 signaling axis, thereby restoring or enhancing T cell function (e.g., proliferation, cytokine production, target cell killing). As used herein, PD-1 axis binding antagonists include PD-1 binding antagonists, PD-L1 binding antagonists, and PD-L2 binding antagonists.

[0217] The term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signal transduction resulting from interference between PD-1 and one or more of its binding partners, e.g., PD-L1 and PD-L2. In some embodiments, a PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to one or more of its binding partners. In particular aspects, a PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and / or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppress, or interfere with signal transduction resulting from the interaction of PD-1 with PD-L1 and / or PD-L2. In one embodiment, the PD-1 binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-1, such that dysfunction of the dysfunctional T cells is reduced (e.g., enhances effector responses to antigen recognition). In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In certain aspects, the PD-1 binding antagonist is MDX-1106 (nivolumab), as described herein. In certain other aspects, the PD-1 binding antagonist is MK-3475 (lambrolizumab), as described herein. In another aspect, the PD-1 binding antagonist is CT-011 (pidilizumab), as described herein. In another aspect, the PD-1 binding antagonist is AMP-224, as described herein.

[0218] The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signaling resulting from the interaction of PD-L1 with one or more of its binding partners, e.g., PD-1, B7-1, etc. In some embodiments, a PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partners. In particular aspects, PD-L1 binding antagonists inhibit the binding of PD-L1 to PD-1 and / or B7-1. In some embodiments, PD-L1 binding antagonists include anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppress, or interfere with signaling resulting from the interaction of PD-L1 with one or more of its binding partners, e.g., PD-1, B7-1, etc. In one embodiment, the PD-L1 binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-L1, such that the dysfunction of dysfunctional T cells is reduced (e.g., enhancing the effector response to antigen recognition). In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In a particular aspect, the anti-PD-L1 antibody is YW243.55.S70, as described herein. In another particular aspect, the anti-PD-L1 antibody is MDX-1105, as described herein. In yet another particular aspect, the anti-PD-L1 antibody is MPDL3280A, as described herein. In yet another aspect, the anti-PD-L1 antibody is MEDI4736, as described herein.

[0219] The term "PD-L2 binding antagonist" refers to a molecule that reduces, blocks, inhibits, suppresses, or interferes with signaling resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In some embodiments, a PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to one or more of its binding partners. In a particular aspect, a PD-L2 binding antagonist inhibits the binding of PD-L2 to PD-1. In some embodiments, PD-L2 antagonists include anti-PD-L2 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and other molecules that reduce, block, inhibit, suppress, or interfere with signaling resulting from the interaction of PD-L2 with one or more of its binding partners, such as PD-1. In one embodiment, the PD-L2 binding antagonist reduces negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes that mediate signaling through PD-L2, such that dysfunctional T cells are made less dysfunctional (e.g., enhance the effector response to antigen recognition). In some embodiments, the PD-L2 binding antagonist is an immunoadhesin.

[0220] A "fixed" or "flat" dose of a therapeutic agent herein refers to a dose administered to a human patient without regard to the patient's body weight (WT) or body surface area (BSA). Thus, a fixed dose or flat dose is expressed as a mg / kg dose or a mg / m 2 It is provided as an absolute amount of therapeutic agent, not as a dose.

[0221] A "loading" dose herein generally comprises an initial dose of a therapeutic agent administered to a patient, followed by one or more maintenance doses thereof. Generally, a single loading dose is administered, although multiple loading doses are contemplated herein. Typically, the amount of the loading dose administered exceeds the amount of the maintenance dose administered, and / or the loading dose is administered more frequently than the maintenance dose, so as to achieve the desired steady-state concentration of the therapeutic agent more quickly than can be achieved with the maintenance dose.

[0222] A "maintenance" dose herein refers to one or more doses of a therapeutic agent administered to a patient over a treatment period. Typically, maintenance doses are administered at spaced treatment intervals, such as about weekly, about every two weeks, about every three weeks, or about every four weeks, preferably every three weeks.

[0223] "Infusion" or "infusing" refers to the introduction of a drug-containing liquid into the body through a vein for therapeutic purposes. Typically, this is accomplished with an intravenous (IV) bag.

[0224] An "intravenous administration bag" or "IV bag" is a bag capable of holding a liquid medication that can be administered through a patient's vein. In one embodiment, the liquid medication is saline (e.g., about 0.9% or about 0.45% NaCl). Optionally, the IV bag is formed from polyolefin or polyvinyl chloride.

[0225] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to an antigen. The variable domains of the heavy and light chains (VH and VL, respectively) of a natural antibody generally have similar structures, and each domain contains four conserved framework regions (FR) and three hypervariable regions (HVR). (See, e.g., Kindt et al., Kuby Immunology, 6 th ed., W.H. Freeman and Co., page 91 (2007). A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a specific antigen may be isolated using the VH or VL domain from an antibody that binds to that antigen to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0226] As used herein, the term "vector" refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors as self-replicating nucleic acid structures as well as vectors that are integrated into the genome of a host cell into which they are introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as "expression vectors."

[0227] A "free cysteine ​​amino acid" refers to a cysteine ​​amino acid residue that is engineered into the parent antibody, that has a thiol functional group (-SH), and that is not paired as an intramolecular or intermolecular disulfide bridge.

[0228] The term "or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, or mixture thereof" is meant to include pharmaceutically acceptable salts, solvates, tautomers, and stereoisomeric forms of the depicted structure. The mixture means that mixtures of these forms can exist; for example, compounds of the present disclosure can include both tautomeric forms and pharmaceutically acceptable salts.

[0229] "Pharmaceutically acceptable" substances refer to those substances that, within the bounds of sound medical judgment, are suitable for use in contact with the tissues of subjects without undue toxicity, irritation, allergic response, etc., commensurate with a reasonable benefit-to-risk ratio, and are effective for their intended use.

[0230] A "pharmaceutical composition" refers to a combination of one or more drug substances and one or more excipients.

[0231] As used herein, "solvate" refers to a complex of variable stoichiometry formed by a solute (e.g., (A), (B), (C), (D) of Formula (1)-(1), or any other compound herein or a salt thereof) and a solvent. Pharmaceutically acceptable solvates can be formed in crystalline compounds where solvent molecules are incorporated into the crystalline lattice during crystallization. The incorporated solvent molecules can be water molecules or non-aqueous molecules such as, but not limited to, ethanol, isopropanol, dimethyl sulfoxide, acetic acid, ethanolamine, and ethyl acetate molecules.

[0232] As used herein, the term "subject" refers to a human or non-human mammal. Examples of non-human mammals include livestock animals such as sheep, horses, cattle, pigs, goats, rabbits, and deer, as well as companion animals such as cats, dogs, rodents, and horses.

[0233] A "therapeutically effective amount" of a drug refers to the amount of a drug or composition that is effective in treating a subject and thus produces the desired therapeutic, ameliorative, inhibitory, or preventative effect. The therapeutically effective amount may depend, inter alia, on the weight and age of the subject and the route of administration.

[0234] "Tolerability" refers to the level of toxicity associated with a therapy or treatment regimen that can be reasonably tolerated by a patient without discontinuing therapy due to toxicity. Non-limiting examples of tolerability include the maximum tolerated dose (MTD).

[0235] "Treating" refers to reversing, alleviating, inhibiting the progression of, or preventing the disorder, disease, or condition to which the term applies, or reversing, alleviating, inhibiting the progression of, or preventing one or more symptoms of, the disorder, disease, or condition.

[0236] "Treatment" refers to the act of "treating," as defined immediately above.

[0237] As used herein, the term "comprising" means "including at least in part of" and is inclusive or open-ended. In interpreting each statement herein that includes the term "comprising," there may be other features, elements, and / or steps present than those preceding the term. Related terms such as "comprising" should be interpreted in the same manner.

[0238] The term "consisting essentially of" limits the claim to the particular materials or steps of the disclosure, "and which do not materially affect the basic and novel characteristics." When the phrase "consisting essentially of" appears in a clause in the body of a claim, rather than immediately following the preamble, it limits only the elements recited in that clause.

[0239] The term "consisting of" excludes any element, step, or ingredient not specified in the claim, and "consisting of" is defined as "closing" the claim to the inclusion of materials other than those recited, except for impurities normally associated therewith. When the phrase "consisting of" appears in a clause in the body of a claim rather than immediately following the preamble, it limits only the elements recited in that clause; other elements are not excluded from the claim as a whole. Although various embodiments herein are presented using the phrase "comprising," it should be understood that, under various circumstances, related embodiments may also be described using the phrase "consisting essentially of" or the phrase "consisting of."

[0240] Purpose The present disclosure finds application in the treatment of proliferative disorders.

[0241] In certain aspects, methods of treating a proliferative disease are provided, the methods comprising administering to a subject a therapeutically effective amount of a compound of formula (I), and salts and solvates thereof, or a composition comprising a compound of formula (I), and salts and solvates thereof.

[0242] In certain embodiments, methods of treating a proliferative disease are provided, the methods comprising administering to a subject a therapeutically effective amount of a conjugate of interest, including a compound of formula (I) and salts and solvates thereof.

[0243] In certain aspects, methods of treating a proliferative disease are provided, the methods comprising administering to a subject a therapeutically effective amount of an antibody-drug conjugate comprising a compound of Formula (I) and salts and solvates thereof.

[0244] The term "proliferative disorder" refers to unwanted or uncontrolled cell proliferation of unwanted, excessive, or abnormal cells, such as neoplastic or hyperplastic growth, whether in vitro or in vivo. Examples of proliferative conditions include, but are not limited to, benign, precancerous, and malignant cell proliferation, including neoplasias and tumors (e.g., histocytoma, glioma, astrocytoma, osteoma), cancers (e.g., lung cancer, small cell lung cancer, hepatocellular carcinoma, gastric cancer or cancer of the stomach region, including gastrointestinal cancer, colorectal cancer, colon cancer, liver cancer, breast cancer, glioblastoma, cervical cancer, ovarian cancer, esophageal (or esophageal) cancer, etc.).

[0013] Proliferative diseases include, but are not limited to, breast cancer, oral cancer, prostate cancer, testicular cancer, liver cancer, rectal cancer, colorectal cancer, endometrial or uterine cancer, uterine cancer, salivary gland cancer, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver carcinoma, anal cancer, penile cancer, head and neck cancer, bladder cancer, pancreatic cancer, brain tumor, sarcoma, osteosarcoma, Kaposi's sarcoma, melanoma), leukemia, psoriasis, bone disease, fibroproliferative disorders (e.g., of connective tissue), and atherosclerosis. Preferably, the proliferative disease is selected from bladder cancer, osteosarcoma, intestinal cancer, brain tumor, breast cancer, cervical cancer, colon cancer, head and neck cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, esophageal cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, renal cancer, retinoblastoma, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, and uterine cancer.

[0245] Any type of cell can be treated, including, but not limited to, bone, eye, head and neck, lung, gastrointestinal (including, for example, mouth, esophagus, large intestine, colon), breast (breast), cervix, ovary, uterus, prostate, liver (hepatic), kidney (renal), bladder, pancreas, brain, and skin.

[0246] One of ordinary skill in the art can readily determine whether a candidate compound treats a proliferative condition for any particular cell type.

[0247] Preferably, the subject is a human, a livestock animal, or a companion animal.

[0248] In a further embodiment, the compound of formula (I), as well as its salts and solvates, can be linked directly or indirectly to a targeting agent (e.g., an antibody, an antibody fragment, a hormone, etc.) to provide a conjugate of interest. The conjugate of interest of the present disclosure can contain one or more compounds of formula (I) (or their salts and solvates). Various conjugates of interest are known in the art and can be used with the compound of formula (I) and its salts or solvates. For example, in certain embodiments, the conjugate of interest is an antibody-drug conjugate, in which one or more compounds of formula (I) are linked directly or indirectly to an antibody. Thus, the compound of formula (I), as well as its salts and solvates, can be used as a payload on the conjugate of interest.

[0249] Preferably, the compound of formula (I) and its salts and solvates for use as a drug in the conjugate of interest are prepared by attaching the compound of formula (I) and its salts and solvates to a targeting agent, either directly or through an optional linker group. Preferably, the compound of formula (I) and its salts and solvates are attached to the targeting agent via a linker group. Preferably, the conjugate of interest is for use in treating a disease, more particularly a proliferative disease. Preferably, the drug can be attached to the targeting agent, either directly or through a linker group, via any suitable functional group contained in the drug. Typically, the drug contains or can be modified to contain one or more functional groups, such as an amine group, a hydroxyl group, or a carboxylic acid group, for attaching the drug to the targeting agent, either directly or through a linker group. In some embodiments, one or more atoms or groups of the compound of formula (I) can be removed during conjugation of the drug to an antibody. In some embodiments, the targeting agent binds to a cell surface receptor or a tumor-associated antigen. In some embodiments, the targeting agent is an antibody. In some embodiments, the targeting agent is a hormone. In some embodiments, the targeting agent is a protein. In some embodiments, the targeting agent is a polypeptide. In some embodiments, the targeting agent is a small molecule (e.g., folic acid).

[0250] The compounds of formula (I) find use as payloads for antibodies or antibody fragments. The compounds of formula (I) allow for easy conjugation to antibodies or antibody fragments or other targeting agents, for example via a linker group.

[0251] Linker Group A linker is a bifunctional compound that can be used to link a drug and a targeting moiety (e.g., an antibody) to form a desired drug conjugate (e.g., an antibody-drug conjugate) or targeting conjugate. Such conjugates are useful for treating diseases because drugs (e.g., cytotoxic agents) can be delivered to cells via antigen recognition.

[0252] In one embodiment, a second section of the linker group is introduced that has a second reactive site (e.g., an electrophilic group) that is reactive with an opposing group (e.g., a nucleophilic group) present on a targeting agent, such as an antibody. Useful nucleophilic groups on antibodies include, but are not limited to, sulfhydryl groups, hydroxyl groups, and amino groups. In this case, the heteroatom of the nucleophilic group of the antibody is reactive with the electrophilic group on the linker group and forms a covalent bond to the linker group. The electrophilic group then provides a linker-payload or linker-drug attachment site, which can include disulfide bridges of the antibody (i.e., stochastic conjugation) or residues (either synthetic or natural) containing electrophilic groups introduced into the antibody to enable efficient conjugation (i.e., site-specific conjugation).

[0253] In another embodiment, the linker group has a reactive site with a nucleophilic group that is reactive with an electrophilic group present on an antibody. Electrophilic groups on an antibody include, but are not limited to, aldehyde groups and ketone carbonyl groups. The heteroatom of the nucleophilic group of the linker group can react with an electrophilic group on the antibody to form a covalent bond to the antibody. Nucleophilic groups in this regard may include, but are not limited to, hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxylate, and arylhydrazide. The electrophilic group on the antibody provides a convenient site for attachment to the linker group. For a more comprehensive list of linking techniques, see Jain, N.; Smith, SW; Ghone, S.; Tomczuk, B., Current ADC Linker Chemistry. Pharmaceutical Research 2015, 32(11), 3526-3540.

[0254] Linkers can be either cleavable or non-cleavable, with cleavable linkers typically represented by a combination of amino acids. Examples of cleavable linkers include, but are not limited to, valine-citrulline, valine-alanine, and any combination of 2 to 8 amino acids. Self-immolative units (e.g., PAB spacers) can be included to aid clean cleavage, and hydrophilic groups (e.g., PEG) can be added to increase the hydrophilicity of the construct. In some embodiments, more preferably, the linker group comprises a self-immolative unit. A range of self-immolative units is known in the art

[30] and has been described, for example, in U.S. Pat. No. 7,754,681 and European Patent Publication No. 0624377.

[0255] Various suitable linker groups are known in the art and can be used as described herein. For example, the maleimide method is routinely used to link antibodies to drug compounds by attaching a linker to the drug with a terminal maleimide group. In addition, methodologies using diarylcyclooctyne moieties (e.g., but not limited to, DBCO, dibenzylcyclooctyne, etc.) are known in the art. Diarylcyclooctynes ​​react with stable azides to provide conjugation through the formation of stable triazoles. Diarylcyclooctynes ​​are thermostable with very narrow and specific reactivity toward azides, resulting in near-quantitative yields of stable triazoles. Furthermore, the reaction does not require the cytotoxic Cu(I) catalyst (toxic to most organisms), thus preventing its use in many biological systems. Furthermore, the alkoxyamine method is also an option in the art. For site-specific conjugation of a drug to an antibody, the antibody may contain a "tag" (which may be proprietary) that reacts with a diarylcyclooctyne (e.g., DBCO), alkyloxyamine, and / or maleimide group to link the antibody to the drug. In some cases, the tag may be a mutant amino acid. Advantageously, linker groups incorporating the various groups described above are available in the art.

[0256] Suitably, the linker group is a L2-R 28 is.

[0257] antibody-drug conjugates Antibody therapy has been established for the targeted treatment of patients with cancer, immune disorders, and angiogenic disorders (Carter, P. (2006) Nature Reviews Immunology 6:343-357). The use of antibody-drug conjugates (ADCs), i.e., immunoconjugates, for the local delivery of cytotoxic or cytostatic agents, i.e., drugs that kill or inhibit tumor cells in the treatment of cancer, targets the delivery of the drug moiety to the tumor and its intracellular accumulation there, whereas systemic administration of these unconjugated drug agents can result in unacceptable levels of toxicity to normal cells (Xie et al (2006) Expert. Opin. Biol. Ther. 6(3):281-291; Kovtun ef a / (2006) Cancer Res. 66(6):3214-3121; Law et al (2006) Cancer Res. 66(4):2328-2337; Wu et al (2005) Nature Biotech. 23(9):1137-1145; Lambert J. (2005) Current Opin. in Pharmacol. 5:543-549; Hamann P. (2005) Expert Opin. Ther. Patents 15(9):1087-1103, Payne, G. (2003) Cancer Cell 3:207-212, Trail ef a / (2003) Cancer Immunol. Immunother.52:328-337, Syrigos and Epenetos (1999) Anticancer Research 19:605-614).

[0258] Maximum efficacy with minimal toxicity is thereby sought. Efforts to design and improve ADCs have focused on the selectivity of monoclonal antibodies (mAbs), as well as drug mechanism of action, drug binding, drug / antibody ratio (loading), and drug release properties (Junutula, et al., 2008b Nature Biotech., 26(8):925-932; Doman et al., (2009) Blood 114(13):2721-2729; U.S. Patent No. 7,521,541; U.S. Patent No. 7,723,485; WO 2009 / 052249; McDonagh (2006) Protein Eng. Design & Sel. 19(7):299-307; Doronina et al., (2006) Bioconj. Chem. 17:114-124; Erickson et al., (2006) Cancer Res. 66(8):1-8; et al. al., (2005) Clin. Cancer Res. 1:843-852, Jeffrey et al., (2005) J. Med. Chem. 48: 1344-1358, Hamblett et al., (2004) Clin. Cancer Res. 10: 7063-7070).

[0259] In some aspects, the present disclosure relates to a compound of Formula (I) and its salts and solvates for use as a drug in an antibody-drug conjugate. In some aspects, the present disclosure relates to an antibody-drug conjugate comprising a compound of Formula (I) and its salts and solvates. Preferably, the compound of Formula (I) and its salts and solvates for use as a drug in an antibody-drug conjugate are prepared by binding the compound of Formula (I) and its salts and solvates to an antibody, either directly or via an optional linker group. Preferably, the compound of Formula (I) and its salts and solvates are bound to an antibody or an antibody fragment thereof via a linker group. Preferably, the antibody-drug conjugate is for use in the treatment of a disease, more particularly a proliferative disease. Preferably, the drug can be bound to the antibody via any suitable functional group contained in the drug, either directly or via a linker group. Typically, the drug contains or can be modified to contain one or more functional groups, such as an amine group, a hydroxyl group, or a carboxylic acid group, for binding the drug to the antibody either directly or via a linker group. In some embodiments, the antibody of the antibody-drug conjugate is an antibody fragment, such as, but not limited to, a single-chain antibody. In some embodiments, one or more atoms or groups of the compound of formula (I) can be removed during binding of the drug to the antibody. In some embodiments, the antibody binds to a cell surface receptor or a tumor-associated antigen.

[0260] In some aspects, the present disclosure relates to the use of a compound of Formula (I) and its salts and solvates as a drug in an antibody-drug conjugate. Preferably, the use of a compound of Formula (I) and its salts and solvates as a drug in an antibody-drug conjugate is achieved by binding the compound of Formula (I) and its salts and solvates to an antibody either directly or via an optional linker group. Preferably, the compound of Formula (I) and its salts and solvates are bound to the antibody via a linker group. Preferably, the antibody-drug conjugate is for use in treating a disease, more particularly a proliferative disease. Preferably, the drug can be bound to the antibody either directly or via a linker group through any suitable functional group contained in the drug. Typically, the drug contains or can be modified to contain one or more functional groups, such as an amine group, a hydroxyl group, or a carboxylic acid group, to bind the drug to the antibody either directly or via a linker group. In some aspects, the antibody of the antibody-drug conjugate is an antibody fragment, such as, but not limited to, a single-chain antibody. In some embodiments, one or more atoms or groups of the compound of formula (I) may be removed during conjugation of the drug to the antibody. In some embodiments, the antibody binds to a cell surface receptor or a tumor-associated antigen.

[0261] In some embodiments, ADCs can be engineered or produced to have (a) an antibody or antigen-binding fragment thereof (e.g., an antibody or antigen-binding fragment thereof) and (b) a compound of Formula (I) or Formula (IV) (i.e., a drug). The drug-to-antibody ratio (DAR) or drug loading refers to the number of drug molecules and / or moieties (i.e., compounds of Formula (I) or Formula (IV) or moieties) conjugated per antibody. In some embodiments, the number of linker-drug moieties attached to an antibody can be any number suitable for ADC development. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody ranges from about 1 to about 10. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 10. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 9. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 8. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 7. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 6. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 5. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 4. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 3. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 2. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is about 1. In some embodiments, the number of compounds of Formula (I) or Formula (IV) or moieties thereof per antibody is more than 4, e.g., 5, 6, 7, 8, 9, 10, 11, 12, or more than 12 linker-drug moieties per antibody.

[0262] Antibodies and antibody fragments The term "antibody" specifically encompasses monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), intact antibodies, and antibody fragments, so long as they exhibit the desired biological activity, e.g., the ability to bind to a desired antigen on a target cell or tissue. Antibodies can be murine, human, humanized, chimeric, or derived from other species. Antibodies are proteins produced by the immune system that are capable of recognizing and binding to a specific antigen. (Janeway, C, Travers, P., Walport, M., Shlomchik (2001) Immuno Biology, 5t h Ed., Garland Publishing, New York). Target antigens generally have multiple binding sites, also called epitopes, recognized by CDRs on antibodies. Each antibody that specifically binds to a different epitope has a different structure. Thus, one antigen can have multiple corresponding antibodies. Antibodies include full-length immunoglobulin molecules or immunoreactive portions of full-length immunoglobulin molecules, i.e., molecules that contain an antigen-binding site that immunospecifically binds to a target or part of an antigen of interest, such targets including, but not limited to, cancer cells or cells that produce autoimmune antibodies associated with autoimmune diseases. The immunoglobulin can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., lgG1, lgG2, lgG3, lgG4, lgA1, and lgA2) or subclass, or allotype (e.g., human G1m1, G1m2, G1m3, non-G1m1 [i.e., any allotype other than G1m1], G1m17, G2m23, G3m21, G3m28, G3m11, G3m5, G3m13, G3m14, G3m10, G3m15, G3m16, G3m6, G3m24, G3m26, G3m27, A2m1, A2m2, Km1, Km2, and Km3) of immunoglobulin molecule. The immunoglobulins may be from any species, including human, murine, or rabbit origin.

[0263] As used herein, "binds to an epitope" is used to mean that an antibody binds to an epitope with higher affinity than a nonspecific partner, e.g., bovine serum albumin (BSA, Genbank Accession Number CAA76847, Version Number CAA76847.1 G1:3336842, Archive Last Updated: January 7, 2011, 2:30 PM). In some embodiments, the antibody binds to an epitope with a ....1 G1:3336842, Archive Last Updated: January 7, 2011, 2:30 PM). In some embodiments, the antibody binds to an epitope with a higher affinity than a nonspecific partner, e.g., bovine serum albumin (BSA, Genbank Accession Number CAA76847.1 G1:3336842, Archive Last Updated: January 7, 2011, 2:30 PM). In some embodiments, the antibody binds to an epitope with a higher affinity than a nonspecific partner, e.g., bovine serum albumin (BSA), Genbank Accession Number CAA76847.1 G1:3336842, Archive Last Updated: January 7, 2011, 2:30 PM). In some embodiments, the antibody binds to an epitope with a higher affinity than a nonspecific partner, e.g., bovine serum albumin (BSA), Genbank Accession Number CAA76847.1 G1:3336842, 4 double, 10 5 double, or 10 6 It binds to the epitope with a 2-fold higher binding constant (Ka).

[0264] The term "antibody fragment" refers to a portion of a full-length antibody, such as its antigen-binding region or its variable region. Examples of antibody fragments include Fab, Fa", F(a"), and scFv fragments; diabodies; linear antibodies; fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies, CDRs (complementary determining regions), single-chain antibody molecules; and multispecific antibodies formed from antibody fragments and epitope-binding fragments of any of the above that immunospecifically bind to a target antigen, e.g., a cancer cell antigen, a viral antigen, or a microbial antigen. As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible natural mutations that may be present in minor amounts. Monoclonal antibodies are highly specific antibodies directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant or epitope on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they may be synthesized uncontaminated by other antibodies. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present disclosure may be made by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). Monoclonal antibodies can also be isolated from phage antibody libraries using the techniques described in Clackson et al (1991) Nature, 352:624-628; Marks et al (1991) J. Mol. Biol., 222:581-597, or from transgenic mice carrying a fully human immunoglobulin system (Lonberg (2008) Curr. Opinion 20(4):450-459).

[0265] As used herein, antibodies, including monoclonal antibodies, refer to antibodies in which a portion of the antibody structure, e.g., the heavy and / or light chain, is identical to or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical to or homologous to corresponding sequences in antibodies derived from another species or belonging to a different antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567 and Morrison et al. (1984) Proc. Natl. Acad. Sci. USA, 81:6851-6855). Chimeric antibodies include "primatized" antibodies containing variable domain antigen-binding sequences derived from a non-human primate (e.g., an Old World monkey or ape) and human constant region sequences. An "intact antibody," as used herein, is one that includes VL and VH domains, as well as a light chain constant domain (CL) and heavy chain constant domains, CH1, CH2, and CH3. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. Intact antibodies may possess one or more "effector functions," which refer to those biological activities attributable to the Fc region of an antibody (a native sequence Fc region or an amino acid sequence variant Fc region). Examples of antibody effector functions include C1q binding, complement dependent cytotoxicity, Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, and down-regulation of cell surface receptors such as the B cell receptor and BCR.

[0266] The antibodies disclosed herein can be modified, for example, to make them less immunogenic to human subjects. This can be accomplished using any of several techniques well known to those skilled in the art, such as humanization.

[0267] Preferably, each targeting agent is independently a protein, a portion of a protein, a polypeptide, a nucleic acid, an antibody, or an antibody fragment. More preferably, each targeting agent is independently an antibody or an antibody fragment. More preferably, each targeting agent is an antibody.

[0268] Suitably, the targeting agent may be any of the antibodies or antibody fragments disclosed herein. Suitably, the targeting agent is an anti-CD22 antibody, an anti-Ly6E antibody, an anti-HER2 antibody, an anti-MUC16 antibody, an anti-STEAP-1 antibody, an anti-NaPi2b antibody, an anti-CD79b antibody, an antibody fragment, a chimeric and humanized antibody, a human antibody, a library-derived antibody, a multispecific antibody, an antibody variant, a substitution, insertion and deletion variant, a glycosylation variant, an Fc region variant, a cysteine ​​engineered antibody variant, or an antibody derivative disclosed herein.

[0269] In various embodiments, the targeting agent may bind to a target selected from acute myeloid leukemia (AML M4) cells, acute promyelocytic leukemia cells, acute lymphoblastic leukemia cells, acute lymphocytic leukemia cells, chronic lymphocytic leukemia cells, chronic myeloid leukemia cells, chronic T-cell lymphocytic leukemia, myelodysplastic syndrome cells, multiple myeloma cells, prostate cancer cells, renal cell adenocarcinoma cells, pancreatic adenocarcinoma cells, lung cancer cells or gastric adenocarcinoma cells, gastric adenocarcinoma cells, breast cancer cells, colon cancer cells, melanoma cells, thyroid cancer cells, ovarian cancer cells, bladder cancer cells, liver cancer cells, head and neck cancer cells, esophageal cancer cells, Hodgkin's lymphoma cells, non-Hodgkin's lymphoma cells, mesothelioma cells, neuroblastoma cells, neuroendocrine tumor cells, neurofibromatosis type 1 (NF1) cells, neurofibromatosis type 2 (NF2) cells, or osteosarcoma cells.

[0270] In some embodiments, the targeting agent binds to a tumor-associated antigen, such as 5-alpha reductase, alpha-fetoprotein, AM-1, APC, APRIL, BAGE, beta-catenin, Bcl12, BCR-ABL, CA-125, CASP-8 / FLICE, cathepin, CD19, CD20, CD21, CD23, CD22, or CD33. CD35, CD44, CD45, CD46, CD5, CD52, CD55, CD59, CDC27, CDK4, CEA, c-MYC, Cox-2, DCC, DcR3, E6 / E7, CGFR, EMBP, Dna78, farnesyltransferase, FGF8b, FGF8a, FLK-1 / KDR, folate receptor, G250, GAGE ​​family, gastrin-17, gastrin-releasing hormone, GD2 / GD3 / GM2, GnRH, GnTV, G P1, gp100 / Pmel17, gp-100-in4, gp15, gp75 / TRP-1, hCG, heparance, Her2 / neu, HMTV, Hsp70, hTERT, IGFR1, IL-13R, iNOS, Ki67, KIAA0205, K-ras, H-ras, N-ras, KSA, LKLR-FUT, MAGE family, mammaglobin, MAP17, melan-A / MART-1, mesothelin, MIC A / B, MT-MMP, mucin, NY-ESO-1, osteonectin, p15, P170 / MDR1, p53, p97 / melanotransferrin, PAI-1, PDGF, uPA, PRAME, probasin, progenipoietin, RAG, PSA, PSA.-1, Rb, RCAS1, SART-1, SSX-family, STAT3, STn, TAG-72, TGF-alpha, TGF-beta, thymosin-beta-15, TNF-alpha, TYRP-, TYRP-2, tyrosinase, VEGF, ZAG, p16INK4, and glutathione-S-transferase.

[0271] Administration and Dosage The compounds of formula (I), formula (IV), and compounds of other formulas described herein (i.e., compounds of formula (I) having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, and compounds of formula (I) having an A-alkylated DNA group of formula (IIIa) or formula (IIIb) as the D moiety) and / or conjugates thereof may be administered alone, in combination with one or more other compounds, or in combination with one or more pharmacologically active compounds different from the compounds of formula (I), formula (IV), and compounds of other formulas described herein (i.e., compounds of formula (I) having a G-alkylated DNA group of formula (II) or one of G1 to G8 as the D moiety, and compounds of formula (I) having an A-alkylated DNA group of formula (IIIa) or formula (IIIb) as the D moiety), and / or conjugates thereof.

[0272] The compounds of the present disclosure can be suitably combined with various ingredients to prepare the compositions of the present disclosure.Preferably, the compositions are combined with pharmaceutically acceptable carriers or diluents to prepare pharmaceutical compositions (can be for human or animal use).Suitable carriers and diluents include isotonic saline, such as phosphate buffered saline.Useful pharmaceutical compositions and methods for their preparation can be found in standard pharmaceutical textbooks.For example, Handbook for Pharmaceutical Additives, 3r d Edition (eds. M. Ash and I. Ash), 2007 (Synapse Information Resources, Inc., Endicott, New York, USA) and Remington: The Science and Practice of Pharmacy, 2 1s Edition (ed. DB Troy) 2006 (Lippincott, Williams and Wilkins, Philadelphia, USA), which are incorporated herein by reference.

[0273] The compounds of the present disclosure can be administered by any suitable route. Preferably, the compounds of the present disclosure will be administered orally or by any parenteral route, usually in the form of a pharmaceutical preparation containing the active ingredient, optionally in the form of a non-toxic organic or inorganic acid or base addition salt in a pharmaceutically acceptable dosage form.

[0274] The compounds of the present disclosure, their pharmaceutically acceptable salts, and pharmaceutically acceptable solvates of either entity can be administered alone, but will generally be administered in admixture with a suitable pharmaceutical excipient diluent or carrier selected with regard to the intended route of administration and standard pharmaceutical practice.

[0275] For example, the compounds of the present disclosure, or salts or solvates thereof, can be administered orally, bucally, or sublingually in the form of tablets, capsules (including soft gel capsules), ovules, elixirs, solutions, or suspensions, which may contain flavorings or coloring agents, for immediate-release, delayed-release, modified-release, sustained-release, controlled-release, or pulsed-delivery applications. The compounds of the present disclosure can also be administered via fast-dispersing or fast-dissolving formulations.

[0276] Such tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate, and glycine, disintegrants such as starch (preferably corn, potato, or tapioca starch), sodium starch glycollate, croscarmellose sodium, and certain complex silicates, and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin, and acacia. Additionally, lubricants such as magnesium stearate, stearic acid, glyceryl behenate, and talc may be included.

[0277] Similar type solid compositions can also be used as filler in gelatin capsules.Preferred excipient in this regard includes lactose, starch, cellulose, milk sugar or high molecular weight polyethylene glycol.For aqueous suspension and / or elixir, the compound of the present disclosure can be combined with various sweeteners or flavorings, coloring materials or dyes, emulsifiers and / or suspending agents, and diluents such as water, ethanol, propylene glycol and glycerin, and combinations thereof.

[0278] Modified-release and pulsatile-release dosage forms may contain excipients, such as those detailed for immediate-release dosage forms, along with additional excipients that act as release rate modifiers, which are coated on and / or incorporated into the body of the device. Release rate modifiers include, but are not limited to, hydroxypropylmethylcellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, polyethylene oxide, xanthan gum, carbomer, ammoniomethacrylate copolymer, hydrogenated castor oil, carnauba wax, paraffin wax, cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, methacrylic acid copolymer, and mixtures thereof. Modified-release and pulsatile-release dosage forms may contain one or a combination of release rate modifiers. Release rate modifiers may be present both within the dosage form, i.e., within the matrix, and / or on the dosage form, i.e., on the surface or coating.

[0279] Fast-dispersing or fast-dissolving dosage forms (FDDF) may contain the following ingredients: aspartame, acesulfame potassium, citric acid, croscarmellose sodium, crospovidone, diascorbic acid, ethyl acrylate, ethyl cellulose, gelatin, hydroxypropyl methylcellulose, magnesium stearate, mannitol, methyl methacrylate, mint flavor, polyethylene glycol, fumed silica, silicon dioxide, sodium starch glycolate, sodium stearyl fumarate, sorbitol, xylitol.

[0280] The compounds of the present disclosure can also be administered parenterally, for example, intravenously, intraarterially, or by infusion techniques. For such parenteral administration, they are best used in the form of a sterile aqueous solution which may contain other substances, for example, sufficient salts or glucose to make the solution isotonic with blood. The aqueous solution should be suitably buffered (preferably to a pH of 3 to 9), if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.

[0281] Preferably, the formulations of the present disclosure are optimized for administration routes such as oral, intravenous, etc.

[0282] Administration can be a single dose, continuous or intermittent (e.g., in divided doses at appropriate intervals) during the course of treatment. Methods for determining the most effective means and dosage are well known to those skilled in the art and will vary depending on the formulation used in therapy, the purpose of therapy, the target cells being treated, and the subject being treated. Single or multiple administrations can be carried out, with the dosage level and administration regimen being selected by the treating physician, veterinarian, or clinician.

[0283] The composition may be administered in various dosages depending on the disorder and patient to be treated and the route of administration. For example, a typical dosage for an adult human may be 100 ng to 25 mg (preferably about 1 μg to about 10 mg) per kg of subject body weight per day.

[0284] Preferably, guidance can be obtained from studies in test animals when estimating initial doses for human subjects. For example, if a specific dose has been identified for mice, preferably the initial test dose for humans would be about 0.5 to 2 times the mg / Kg value given to mice.

[0285] Other forms Unless otherwise specified, the above includes known ionic, salt, solvate, and protected forms of these substituents. For example, a reference to carboxylic acid (-RCOOH) also includes the anionic (carboxylic acid ester) form (-RCOO). - ), a salt or solvate thereof, as well as standard protected forms. Similarly, a reference to an amino group includes the protonated form (-RN + HR 1 R 2 ), salts or solvates of the amino group, such as hydrochlorides, as well as standard protected forms of the amino group. Similarly, a reference to a hydroxyl group also includes the anionic form (—O - ), including salts or solvates thereof, as well as standard protected forms.

[0286] Isomers, salts and solvates Certain compounds may exist in one or more particular geometric, optical, enantiomeric, diastereomeric, epimeric, atropic, mesomeric, tautomeric, conformational, or anomeric forms, including, but not limited to, cis and trans, E and Z, c-, t-, and r-, endo-, exo-, R-, S-, and meso-, D and L, d and l, (+) and (−) forms, keto-, enol-, and enolate forms, syn- and anti-, synclinal-, and anticlinal forms, alpha- and beta-, axial-, equatorial-, boat-, chair-, twisted-, enveloped-, and half-chair forms, and combinations thereof, hereinafter collectively referred to as “isomers” (or “isomeric forms”).

[0287] It should be noted that, except as tautomeric forms are discussed below, the term "isomer," as used herein, specifically excludes structural (or constitutional) isomers (i.e., isomers that differ in the bonding between their atoms and not simply in the position of their atoms in space). For example, a reference to a methoxy group, -OCH, should not be construed as a reference to its structural isomer, a hydroxymethyl group, -CHOH.

[0288] Reference to a structural class may well include structural isomeric forms that fall within that class (e.g., C 1-7 Alkyl includes n-propyl and isopropyl, butyl includes n-, iso-, sec- and tert-butyl, methoxyphenyl includes ortho-, meta- and para-methoxyphenyl).

[0289] The above exclusion does not apply to tautomers, e.g., keto-, enol-, and enolate-forms, as in, for example, the following tautomeric pairs: keto / enol, imine / enamine, amide / iminoalcohol, amidine / amidine, nitroso / oxime, thioketone / enethiol, N-nitroso / hydroxyazo, and nitro / aci-nitro.

[0290] In particular, it should be noted that the term "isomer" includes compounds with one or more isotopic substitutions. For example, H is 1 H, 2 H(D), and 3 H(T) and C can be in any isotopic form, including 12 C. 13 C, and 14 C can be any isotopic form, including O 16 O and 18 It can be any isotopic form containing O, etc.

[0291] Unless otherwise specified, a reference to a particular compound includes all such isomeric forms (in whole or in part), including racemates and other mixtures thereof.

[0292] Methods for the preparation (e.g., asymmetric synthesis) and separation (e.g., XXX orbidize XXX ion fractionation and chromatographic means) of such isomeric forms are known in the art or are readily obtained by adapting the methods taught herein or known methods in a known manner.

[0293] Unless otherwise specified, a reference to a particular compound also includes ions, salts, solvates, and protected forms thereof, for example, as discussed below.

[0294] In some embodiments, the compounds of formula (I) and salts and solvates thereof include pharmaceutically acceptable salts of compounds of formula (I).

[0295] Compounds of formula (I), including those specifically named above, can form pharmaceutically acceptable complexes, salts, solvates and hydrates, including non-toxic acid addition salts (including diacids) and base salts.

[0296] If the compound is cationic or has a functional group that can be cationic (e.g., -NH2 can be converted to -NH3 +Acid addition salts may be formed with suitable anions. Examples of suitable inorganic anions include, but are not limited to, those derived from the following inorganic acids: hydrochloric acid, nitric acid, nitrous acid, phosphoric acid, sulfuric acid, sulfurous acid, hydrobromic acid, hydroiodic acid, hydrofluoric acid, phosphoric acid, and phosphorous acid. Examples of suitable organic anions include, but are not limited to, those derived from the following organic acids: 2-acetyoxybenzoic, acetic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucheptonic, gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic, benzenesulfonic, propionic, pyruvic, salicylic, stearic, succinic, sulfanilic, tartaric, toluenesulfonic, and valeric acids. Examples of suitable polymeric organic anions include, but are not limited to, those derived from the following polymeric acids: tannic acid, carboxymethyl cellulose.These salts include acetate, adipate, aspartate, benzoate, besylate, bicarbonate, carbonate, bisulfate, sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hybenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, and isethionate. , lactate, malate, maleate, malonate, mesylate, methylsulfonate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate, hydrogen phosphate, dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinofoate.

[0297] For example, if the compound is anionic or has a functional group that can be anionic (e.g., -RCOOH becomes -RCOO), - Base salts can be formed with suitable cations. Examples of suitable inorganic cations include, but are not limited to, metal cations such as alkali or alkaline earth metal cations, ammonium and substituted ammonium cations, as well as amines. Examples of suitable metal cations include sodium (Na + ), potassium (K + ), magnesium (Mg 2+ ), calcium (Ca 2+ ), zinc (Zn 2+ ), and aluminum (Al 3+ ) Examples of suitable organic cations include ammonium ions (i.e., NH4 + ) and substituted ammonium ions (e.g., NHR + , NH2R2 + , NHR3 + , NR4 +) are substituted ammonium ions, but are not limited to those derived from ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, as well as amino acids such as lysine and arginine. Examples of common quaternary ammonium ions include N(CH3)4 + Examples of suitable amines include arginine, N'-dibenzylethylene-diamine, chloroprocaine, choline, diethylamine, diethanolamine, dicyclohexylamine, ethylenediamine, glycine, lysine, N-methylglucamine, olamine, 2-amino-2-hydroxymethyl-propane-1,3-diol, and procaine. For a discussion of useful acid addition and base salts, see S. M. Berge et al., J. Pharm. Sci. (1977) 66:1-19; also see Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (2011).

[0298] Pharmaceutically acceptable salts can be prepared using various methods. For example, the compound of formula 1 can be reacted with an appropriate acid or base to obtain the desired salt. The precursor of the compound of formula I can also be reacted with an acid or base to remove acid-labile or base-labile protecting groups or to open the lactone or lactam group of the precursor. Furthermore, the salt of the compound of formula 1 can be converted into another salt by treating with an appropriate acid or base or by contacting with an ion exchange resin. After the reaction, the salt can then be isolated by filtration if it precipitates from solution, or by evaporation to recover the salt. The degree of ionization of the salt can vary from completely ionized to almost non-ionized.

[0299] It may be convenient or desirable to prepare, purify, and / or handle a corresponding solvate of an active compound. The term "solvate" describes a molecular complex comprising a compound and one or more pharmaceutically acceptable solvent molecules (e.g., EtOH). The term "hydrate" refers to a solvate in which the solvent is water. Pharmaceutically acceptable solvates include those in which the solvent may be isotopically substituted (e.g., DO, acetone-d6, DMSO-d6).

[0300] The currently accepted classification system for solvates and hydrates of organic compounds distinguishes between isolated moiety, channel, and metal ion-coordinated solvates and hydrates. See, for example, KR Morris (HGBrittain ed.) Polymorphism in Pharmaceutical Solids (1995). Isolated moiety solvates and hydrates are those in which the solvent (e.g., water) molecules are isolated from direct contact with each other by intervening molecules of the organic compound. In channel solvates, the solvent molecules are in lattice channels next to other solvent molecules. In metal ion-coordinated solvates, the solvent molecules are bound to the metal ion.

[0301] When the solvent or water is tightly bound, the complex will have a well-defined stoichiometry independent of humidity. However, when the solvent or water is weakly bound, such as in solvates of channels and in hygroscopic compounds, the water or solvent content will depend on humidity and drying conditions. In such cases, non-stoichiometry will typically be observed.

[0302] Further particular and preferred aspects are set out in the accompanying independent and dependent claims. Features from the dependent claims may be combined with features of the independent claims as appropriate and in combinations other than those explicitly set out in the claims.

[0303] Synthesis strategy Compounds of formula (I) can be prepared using the techniques described below. Some of the schemes and examples may omit details of conventional reactions, such as oxidation, reduction, separation techniques (extraction, evaporation, precipitation, chromatography, filtration, trituration, crystallization, etc.), and analytical procedures known to those skilled in the art of organic chemistry. Details of such reactions and techniques can be found in Richard Larock, Comprehensive Organic Transformations, A Guide to Functional Group Preparations, 2n d Ed (2010), and the multi-volume series Compendium of Organic Synthetic Methods (1974 and later), edited by Michael B. Smith et al., among many other publications. Starting materials and reagents may be obtained from commercial sources or prepared using literature methods. Some reaction schemes may omit minor products resulting from chemical transformations (e.g., alcohols from the hydrolysis of esters, CO from the decarboxylation of diacids, etc.). Furthermore, in some instances, reaction intermediates may be used in subsequent steps without isolation or purification (i.e., in situ).

[0304] In some of the reaction schemes and examples that follow, certain compounds can be prepared using protecting groups, which prevent undesired chemical reactions at otherwise reactive sites. Protecting groups may also be used to enhance the solubility of the compound or otherwise modify its physical properties. For a description of protecting group strategies, materials and methods for introducing and removing protecting groups, and a collection of useful protecting groups for common functional groups, including amines, carboxylic acids, alcohols, ketones, aldehydes, and the like, see T.W. Greene and P.G. Wuts, Protecting Groups in Organic Chemistry, 4t h Edition, (2006) and P. Kocienski, Protective Groups, 3r d Edition (2005).

[0305] Generally, the chemical transformations described throughout this specification can be carried out using substantially stoichiometric amounts of the reactants, although certain reactions can benefit from using one or more of the reactants in excess. Additionally, although many of the reactions disclosed throughout this specification can be carried out at about room temperature (RT) and ambient pressure, some reactions may be carried out at elevated pressures or employ higher temperatures (e.g., reflux conditions) or lower temperatures (e.g., −78° C. to 0° C.), depending on reaction rate, yield, etc. All references in this disclosure to stoichiometric ranges, temperature ranges, pH ranges, etc., include both indicated endpoints, whether or not explicitly referred to as “ranges.”

[0306] Many chemical transformations may also employ one or more compatible solvents, which may affect reaction rate and yield. Depending on the nature of the reactants, the one or more solvents may be polar protic solvents (including water), polar aprotic solvents, nonpolar solvents, or some combination. Representative solvents include saturated aliphatic hydrocarbons (e.g., n-pentane, n-hexane, n-heptane, n-octane), aromatic hydrocarbons (e.g., benzene, toluene, xylene), halogenated hydrocarbons (e.g., methylene chloride, chloroform, carbon tetrachloride), aliphatic alcohols (e.g., methanol, ethanol, propan-1-ol, propan-2-ol, butan-1-ol, 2-methyl-propan-1-ol, butan-2-ol, 2-methyl-propan-2-ol, pentan-1-ol, 3-methyl-butan-1-ol, hexane-1-ol, 2-methoxy-ethanol, 2-ethoxy-ethanol, 2-butoxy-ethanol, 2-(2-methoxy-ethoxy)-ethanol, 2-(2-ethoxy-ethoxy)-ethanol, 2-(2-butoxy-ethoxy)-ethanol, ethanol), ethers (e.g., diethyl ether, diisopropyl ether, dibutyl ether, 1,2-dimethoxyethane, 1,2-diethoxyethane, 1-methoxy-2-(2-methoxy-ethoxy)-ethane, 1-ethoxy-2-(2-ethoxy-ethoxy)-ethane, tetrahydrofuran, 1,4-dioxane), ketones (e.g., acetone, methyl ethyl ketone), esters (methyl acetate, ethyl acetate), nitrogen-containing solvents (e.g., formamide, N,N-dimethylformamide, acetonitrile, N-methyl-pyrrolidone, pyridine, quinoline, nitrobenzene), sulfur-containing solvents (e.g., carbon disulfide, dimethyl sulfoxide, tetrahydro-thiophene-1,1,-dioxide), and phosphorus-containing solvents (e.g., HMPA, hexamethylphosphoramide).

[0307] One synthetic strategy for adding a monosaccharide unit to a compound of formula I may include the following (stereochemistry not shown): Glucose-based N11-carbamate [ka] Monosaccharide units linked to either a primary alcohol (right) or a secondary alcohol (left: any of the four groups), or Glucuronides [ka] either carbamates (left, either alcohol groups) or directly bonded amides (right).

[0308] In some aspects, the present disclosure relates to compounds of formula (I) and their salts and solvates for use in preparing a drug in an antibody-drug conjugate. Preferably, when compounds of formula (I) and their salts and solvates contain one or more functional groups (such as amine, hydroxyl, or carboxylic acid groups) for linking a drug to an antibody either directly or via a linker group, the compounds of formula (I) and their salts and solvates can be used directly to prepare an antibody-drug conjugate. Preferably, compounds of formula (I) and their salts and solvates can be used in preparing an antibody-drug conjugate by being modified to contain one or more functional groups (such as amine, hydroxyl, or carboxylic acid groups) for linking a drug to an antibody either directly or via a linker group. Preferably, compounds of formula (I) and their salts and solvates can be used in preparing an antibody-drug conjugate by being modified to contain one or more antibody linker groups, in which an antibody is linked to a drug via one or more antibody linker groups. Thus, the present disclosure provides compounds of Formula (I) further comprising one or more antibody linker groups. Preferably, compounds of Formula (I) may contain one, two, or three antibody linker groups. Preferably, compounds of Formula (I) may contain one or two antibody linker groups. Preferably, compounds of Formula (I) may contain one antibody linker group. In some embodiments, one or more atoms or groups of compounds of Formula (I) may be removed during attachment of a drug to an antibody, or attachment of an antibody linker to a drug, or during modification of a drug containing one or more functional groups (such as an amine, hydroxyl, or carboxylic acid group) to attach a drug to an antibody either directly or via an antibody linker group.

[0309] Various suitable antibody linker groups are known in the art and can be used as described herein. For example, the maleimide method is routinely used as a method for linking antibodies to drug compounds by attaching an antibody linker with a terminal succinimide group to the drug (forming a succinimide-antibody conjugate). Furthermore, methodologies using diarylcyclooctyne moieties (e.g., but not limited to, DBCO, dibenzylcyclooctyne, etc.) are also options used in the art. Diarylcyclooctynes ​​react with azides to provide conjugation via the formation of stable triazoles. Diarylcyclooctynes ​​are thermostable with very narrow and specific reactivity toward azides, resulting in near-quantitative yields of stable triazoles. Furthermore, the reaction does not require the cytotoxic Cu(I) catalyst (toxic to most organisms), thus preventing its use in many biological systems. Furthermore, the alkoxyamine method is also an option used in the art. For site-specific conjugation of a drug to an antibody, the antibody may contain a "tag" (which may be proprietary) that reacts with dairylcyclooctyne (e.g., DBCO), alkyloxyamine, and / or maleimide groups to link the antibody to the drug. In some instances, the tag may be a mutated amino acid. Suitable antibody linker groups incorporating the various groups described above are commercially available in the art.

[0310] The present disclosure will be further described in the following embodiments, which do not limit the scope of the disclosure as set forth in the claims.

[0311] Embodiment 1. A compound of formula (I): DQBT (I) or a salt, solvate, or tautomer thereof, wherein: D is a source of alkylated DNA minor groove binding units, Q is a linker, B is a DNA-binding amide-containing chain, T is a terminal group, wherein D, B, Q and / or T comprise at least one carbohydrate substituent. Embodiment 2. The sugar chain substituent is R S and preferably R S is glycosyl or O-glycosyl. Embodiment 3. D comprises a G-alkylated DNA group of formula (II): [ka] During the ceremony, The dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4; The wavy line indicates the point of attachment to Q; m is 0 or 1, R1, R3, and R4 are independently selected from H and R 29 and R2 is selected from H, L2-R 28、 R 29 , and -L S -R S or one of R1 and R2, R2 and R3, or R3 and R4, together with the carbon atom to which they are attached, is selected from a 6-membered aryl, or one, two, or three independently selected optional R 20 forming a 5- or 6-membered cyclic, heterocyclic or heteroaryl ring optionally substituted by a group, R5 and R6 are: (i) R5 is H, OH, and OC 1-6 alkyl, and R6 is selected from H, SO3H, -L S -R S , nitrogen protecting group, -L2-R 28 and R A or (ii) R5 is oxo or H and R6 is H or C 1-6 alkyl, or (iii) R5 and R6 together form a double bond; R7 and R9 are independently H and R 20 is selected from R8 is H, SR 24 , SCH2Ph, R 20 , L2-R 28 , and -L S -R S is selected from R A is (CH2) j -OH, (CH2) j -CO2R 26 , C(=O)-O-(CH2) k -NR 26 R 27 , (CH2) j NR 26 R 27 , C(=O)-NH-(CH2) j -NR 26 R 27 and C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 is selected from L2 is a bond or a linker moiety having 1 to 200 non-hydrogen atoms selected from C, N, P, O, S or halogen, and may incorporate an ether, oxo, carboxamidyl, uretanyl, branched, cyclic, unsaturated, heterocyclyl, aryl, or heteroaryl moiety; R 28 are azides, alkynes, bisulfones, carbohydrazides, hydrazines, hydroxylamines, iodoacetamides, isothiocyanates, maleimides, phosphines, pyridopyridazines, semihydrazides, succinimidyl esters, sulfodichlorophenol esters, sulfonyl halides, sulfosuccinimidyl esters, 4-sulfotetrafluorophenyl esters, tetrafluorophenyl esters, thiazoles, R A , O-(CH2) k -NR 26 R 26, NHNH2, or a targeting agent selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; Each R 29 are independently, R 20 , R 21 , =CH2, =CH-(CH2) s -CH3, =CH-(CH2) s -R 21 , =O, (CH2) s -OR 21 , (CH2) s -CO2R 21 , (CH2) s -NR 21 R 24 , O-(CH2) t -NR 21 R 24 , NH-C(O)-R 21 , O-(CH2) t -NH-C(O)-R 21 , O-(CH2) t -C(O)-NH-R 21 , (CH2) s -SO2R 21 , O-SO2R 21 , (CH2) s -C(O)R 21 and (CH2) s -C(O)NR 21 R 24 is selected from Each R 20 are independently F, Cl, Br, (CH2) j -OH, C 1-6 Alkyl, OC 1-6 Alkyl, OCH2Ph, (CH2) j -CO2R 26 , O-(CH2) k -NR 26 R 27 , C(=O)-O-(CH2) k -NR 26 R 27 , C(=O)-NR 26 R 27 , (CH2) j -NR 26 R 27 , N.R.26 NH2, C(=O)-NH-(CH2) j -NR 26 R 27 , C(=O)-NH-C6H4-(CH2) j -R 26 , C(=O)-NH-(CH2) k -C(=NH)NR 26 R 27 , -L2-R 28 , S(O)2-(C 1-6 alkyl), O-(CH2) k -O-(C 1-6 alkyl), (CH2) j -S(O)2-NR 26 R 27 , C(=NH)-O-(C 1-6 alkyl), (CH2) k -O-(C 1-6 alkyl), CN, NCO, Cy, C(O)-NH-(CH2) j -Cy, C(O)-Cy, NH-C(O)-NR 26 R 27 is selected from [ka] each j and s is independently selected from 0, 1, 2, 3, 4, 5, or 6; each k and t is independently selected from 1, 2, 3, 4, 5, or 6; Each R 21 are independently H, C 1-12 Alkyl, C 5-6 Heterocyclyl, C 5-9 Heteroaryl, C 6-15 Heteroarylalkyl, phenyl, and C 7-12 and aralkyl groups, wherein the heterocyclyl, heteroaryl, heteroarylalkyl, phenyl, and aralkyl groups are selected from one, two, or three independently selected optional R 20 may be substituted with a group, Each R 24 , R 26 , and R 27 are independently H and C 1-12 alkyl, Each Cy independently represents C 5-6 Heterocyclyl or C 5-6 heteroaryl groups, wherein the heterocyclyl or heteroaryl group is selected from one or two R 20 may be substituted with a group, L S is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 - and these chains are P, O, S, NH, C 5-9 which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; 5-9 The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L S teeth, [ka] and L C is an amino acid, an amino acid derivative, a peptide chain having 2 to 6 amino acids or amino acid derivatives, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-8 -, and these chains contain one or more groups selected from one or more P, O, S and / or NH groups, and / or C 5-9 may be interrupted by heteroarylene and / or phenylene, and each C 5-9 the heteroarylene group and / or each phenylene group may be optionally substituted; R SThe compound of embodiment 1 or 2, wherein is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl. Embodiment 4. The compound of embodiment 3, wherein G is selected from the group of formulas G1-G8. Embodiment 5. The compound of embodiment 1 or 2, wherein D comprises an A that is an A-alkylated DNA group of formula (IIIa) or (IIIb). Embodiment 6. The compound of embodiment 3 or 4, wherein R1, R3, R7, and R9 are each H. Embodiment 7. The compound of embodiment 3 or 4, wherein the compound is a compound of formula IV. Embodiment 8. Q includes X1-L-X2, During the ceremony, X1 is O, S, NR 13 , C.R. 13 R 14 , C.R. 13 R 14 O, C(=O), C(=O)NR 13 , N.R. 13 selected from C(═O), OC(O), and C(O)—O, or absent; L is an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-6 -, these chains being selected from one or more P, O, S and / or NH groups, and / or C 5-9 may be interrupted by heteroarylene and / or phenylene, and each C 5-9 the heteroarylene group and / or each phenylene group may be optionally substituted; X2 is O, S, NR 15 , C.R. 15 R 16 , C.R. 15 R 16 O, C(=O), C(=O)NR15 , N.R. 15 selected from C(═O), OC(O), and C(O)—O, or absent; R 13 , R 14、 R 15 , and R 16 are independently H and C 1-6 The compound of any one of embodiments 1 to 7, wherein the alkyl is selected from: Embodiment 9. B is (A) q Including, During the ceremony, q is selected from 0, 1, 2, 3, 4, 5, and 6; A is selected from the following: [ka] For each A group, one of Y and Y is independently NR 30 , S, and O; the other of Y3 and Y4 is CH; and Y5 is independently selected from CR 30 , N, S, and COH; For each A2 group, one of Y6 and Y7 is independently selected from N and CH, and the other of Y6 and Y7 is CR 30 and Each R 30 are independently H, C 1-6 Alkyl, L2-R 28 , and R S 9. The compound of any one of embodiments 1 to 8, selected from: Embodiment 10. T includes groups of the formula: [ka] During the ceremony, p is 0 or 1, R T -L2-R 28 , phenyl, and C 5-9 heteroaryl, selected from phenyl and C 5-9The heteroaryl group optionally includes OH, C 1-6 Alkyl, OC 1-6 Alkyl, -L2-R 28 , (CH2) j -CO2R 11 , O-(CH2) k -NR 11 R 12 , (CH2) j -NR 11 R 12 , C(=O)-NH-(CH2) k -NR 11 R 12 , C(=O)-NH-R 24 , and C(=O)-NH-(CH2) k -C(=NH)NR 11 R 12 and optionally, but optionally substituted with up to three optional substituents selected from 5-9 heteroaryl is other than indolyl; R 19 is H, C 1-6 Alkyl, L2-R 28 , R S , and (CH2) t -NR 20 R 21 is selected from Y1 and Y2 are independently N or CR 31 and at least one of Y1 and Y2 is CR 31 and Each R 31 are independently H, C 1-6 Alkyl, L2-R 28 , and R S is selected from R 11 , R 12 , and R 24 are independently H, -L2-R 28 , and C 1-6 The compound of any one of embodiments 1 to 9, wherein the alkyl is selected from: Embodiment 11. The compound is selected from compounds of the following formula: [ka] [ka] [ka] [ka] [ka] [ka] [ka] In the formula, R S 10. Compounds of formula (I) and salts and solvates thereof according to any one of the preceding claims, wherein is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl. Embodiment 12. The compound has at least one L2-R 28 12. The compound of any one of embodiments 1 to 11, or a pharmaceutically acceptable salt thereof, comprising a group. Embodiment 13. D, T, Q and / or B are L2-R 28 13. The compound of embodiment 12, or a pharmaceutically acceptable salt thereof, substituted with a group. Embodiment 14. L2 is [ka] is selected from In the formula, X AA is the amino acid sequence, and -K2 is -[CH2CH2O- 0-50 - or -[CH2] 0-12 14. The compound of embodiment 12 or 13, or a pharmaceutically acceptable salt thereof, wherein Embodiment 15. L S is: [ka] In the formula, L C is an amino acid, an amino acid derivative, a peptide chain having 2 to 6 amino acids or amino acid derivatives, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH2) 1-12 -, polyethylene glycol chain -(OCH2CH2) 1-8 -, and these chains contain one or more groups selected from one or more P, O, S and / or NH groups, and / or C 5-9 may be interrupted by heteroarylene and / or phenylene, and each C 5-9 The compound of any one of embodiments 3 to 14, or a pharmaceutically acceptable salt thereof, wherein the heteroarylene group and / or each phenylene group is optionally substituted. Embodiment 16. L C teeth, [ka] at will [ka] 16. The compound of embodiment 15, comprising: Embodiment 17. L C is a polyethylene glycol chain -(OCH2CH2) 1-8 17. The compound of embodiment 15 or 16, optionally comprising -(OCH2CH2)8-. Embodiment 18. R 28 -L C 18. The compound of any one of embodiments 15 to 17, wherein: [ka] Embodiment 19. X AA 19. The compound of any one of embodiments 12 to 18, or a pharmaceutically acceptable salt thereof, wherein is L-valyl-L-alanine. Embodiment 20. R 28 is the following maleimide: [ka] 20. The compound of any one of embodiments 12 to 19, or a pharmaceutically acceptable salt thereof, optionally linked to a targeting agent. Embodiment 21. L2-R 28 teeth, [ka] [ka] Contains or [ka] Contains or [ka] Contains, or [ka] Including, 21. The compound of any one of embodiments 12 to 20, or a pharmaceutically acceptable salt thereof, optionally linked to a targeting agent. Embodiment 22. The compound of formula (I), as well as salts and solvates thereof, according to any one of embodiments 21 to 17, which is linked, either directly or indirectly, to a targeting agent to provide the conjugate of interest. Embodiment 23. The compound has at least one L2-R 28 and the targeting agent comprises an L2-R 28 The compound of formula (I), as defined in embodiment 22, and salts and solvates thereof, linked to the compound via a group. Embodiment 24. 24. The compound of any one of embodiments 20 to 23, wherein the targeting agent comprises an antibody, an antibody fragment, a hormone, or a hormone fragment. Embodiment 25. 25. A compound of formula (I), and salts and solvates thereof, according to any one of embodiments 1 to 24, for use as a medicament. Embodiment 26. 25. Compounds of formula (I), and salts and solvates thereof, as defined in any one of embodiments 1 to 24, for use in the treatment of a proliferative disease. Embodiment 27. Compounds of formula (I), as defined in embodiment 26, and salts and solvates thereof, for use in the treatment of a proliferative disease, wherein the proliferative disease is selected from bladder cancer, osteosarcoma, intestinal cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, esophageal cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, retinoblastoma, sarcoma, skin cancer, cancer of the stomach, testicular cancer, thyroid cancer, and uterine cancer. Embodiment 28. A pharmaceutical composition comprising a compound of formula (I) as defined in any one of embodiments 1 to 24, and salts and solvates thereof, and a pharmaceutically acceptable excipient, carrier or diluent. Embodiment 29. The use of compounds of formula (I), and salts and solvates thereof, as defined in any one of embodiments 1 to 28, in the manufacture of a medicament for treating a proliferative disease. Embodiment 30. A method for treating a patient suffering from a proliferative disease, comprising administering to the patient a therapeutically effective amount of a compound of any one of embodiments 1 to 24 or a pharmaceutical composition of embodiment 28. Embodiment 31. 31. The method of embodiment 30, wherein the proliferative disease is selected from bladder cancer, osteosarcoma, intestinal cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, esophageal cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, retinoblastoma, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, and uterine cancer. Embodiment 32. An antibody-drug conjugate comprising a compound of formula (I) as defined in any one of embodiments 1 to 24, and salts and solvates thereof.

[0312] Further particular and preferred aspects are set out in the accompanying independent and dependent claims. Features from the dependent claims may be combined with features of the independent claims as appropriate and in combinations other than those explicitly set out in the claims. [Example]

[0313] Example 1: Synthesis of A-Alkylating Agent Compounds, PDD-C8 Substituted Compounds, PDD-N11 Substituted Compounds, and Control Compounds General Synthetic Methods. All reagents and solvents were purchased from standard commercial sources and used as purchased. Anhydrous reactions were carried out under an inert atmosphere of argon using anhydrous solvents that were used as purchased without further drying. Thin-layer chromatography (TLC) was performed on silica gel aluminum plates (Merck 60, F 254 ) and flash column chromatography was performed using a Biotage Isolera One (automated flash chromatography system) with TLC monitoring (UV, 254 nm).

[0314] All nuclear magnetic resonance (NMR) spectra were acquired at room temperature using a Bruker 600 MHz Ultrashield (Bruker Avance NEO console with cryoplatform) or a Varian Mercury Vx Agilent 400 MHz spectrometer, for which chemical shifts are expressed in ppm relative to the solvent, and coupling constants are expressed in Hz. Microwave reactions were performed on a Biotage Initiator+ microwave synthesizer. High-resolution mass spectrometry (HRMS) was performed on a Thermo Scientific Exactive HCD Orbitrap mass spectrometer. Yields refer to isolated material (homogeneous by TLC and NMR) unless otherwise stated, and names are assigned according to IUPAC nomenclature.

[0315] Liquid chromatography-mass spectrometry (LCMS) analytical methods A–C were performed on a Waters Alliance 2695 column using water (A) and acetonitrile (B) as the mobile phase. Formic acid (0.1%) was added to both the acetonitrile and water to ensure acidic conditions throughout the analysis. Functional type: diode array (535 scans). Column type: monolithic C18 50 × 4.60 mm. Mass spectrometry data were collected using a Waters Micromass ZQ instrument interfaced with an HPLC equipped with a Waters 2996 PDA. The Waters Micromass ZQ parameters used were: capillary (kV), 3.38; cone (V), 35; extractor (V), 3.0; source temperature (°C), 100; desolvation temperature (°C), 200; cone flow rate (L / h), 50; and desolvation flow rate (L / h), 250. The gradient conditions are as follows:

[0316] Method A (10 min): 95% A / 5% B to 50% B over 3 min. Then 50% B to 80% B over 2 min. Then 80% B to 95% B over 1.5 min and held constant for 1.5 min. This was then reduced to 5% B over 0.2 min and maintained at 5% B for 1.8 min. The flow rate was 0.5 mL / min, and 200 μL was separated through a zero-dead-volume T-piece and passed through the mass spectrometer. The UV detector wavelength range was 220-400 nm.

[0317] Method B (5 min): 95% A / 5% B to 90% B over 3 min. Then, 90% B to 95% B over 0.5 min and held constant for 1 min. This was then reduced to 5% B over 0.5 min. The flow rate was 1.0 mL / min, and 100 μL was separated through a zero-dead-volume T-piece and passed through the mass spectrometer. The UV detector wavelength range was 220-500 nm.

[0318] Method C (5 min): 95% A / 5% B, ramped to 90% B over 3 min and then to 95% B over 0.5 min. The gradient was then held at 95% B for 1 min and ramped back to 5% B over 0.5 min. The total run duration was 5 min, the solvent flow rate was 1 mL / min, and 100 μL was separated through a zero-dead-volume T-piece and passed through the mass spectrometer. The UV detector wavelength range was 220–500 nm.

[0319] Liquid chromatography mass spectrometry (LCMS) analysis methods D–G were performed using a Shimadzu LC-20AD series binary pump and diode array detector. Column type: Agilent Poroshell 120 EC-C18, 2.7 μm, 4.6 × 50 mm. Mobile phase: A: 0.05% formic acid in water (v / v), B: 0.05% formic acid in acetonitrile (v / v). Flow rate: 1 mL / min at 25 °C. Detectors: 214 nm, 254 nm. Gradient stop time: 5 min. MS: 2020, quadrupole LC / MS, ion source: API-ESI, TIC: 100–1300 m / z, drying gas flow rate: 15 L / min, nebulizer pressure: 1.5 L / min, drying gas temperature: 250 °C, Vcap: 4500 V. Sample preparation: The sample was dissolved in methanol at 1-10 μg / mL and then filtered through a 0.22 μm filter membrane. Injection volume: 1-10 μL. The gradient conditions are as follows:

[0320] Method D (5 min): 20% A / 80% B for 0.5 min, increased to 100% B over 3.5 min, then held at 100% B for 0.5 min, then increased back to 20% A / 80% B for 0.5 min.

[0321] Method E (5 min): 50% A / 50% B for 0.5 min, increased to 100% B over 3.5 min, then held at 100% B for 0.5 min, then increased back to 50% A / 50% B for 0.5 min.

[0322] Method F (5 min): 85% A / 15% B for 0.5 min, increased to 100% B over 3.5 min, then held at 100% B for 0.5 min, then increased back to 85% A / 15% B for 0.5 min.

[0323] Method G (5 min): 97% A / 3% B for 0.5 min, increased to 30% A / 70% B over 3.5 min, then increased to 100% B over 0.5 min, then increased back to 97% A / 3% B over 0.5 min.

[0324] Optical rotations were measured with an SGWzz-1 automatic polarimeter (Shanghai Shen Guang Instrument Co., Ltd.) or a Bellingham-Stanley ADP 440+ polarimeter.

[0325] Reverse-phase preparative HPLC was performed on a Shimadzu LC equipped with a CTC IFC using a Phenomenex Gemini NX 5m, C18, 110Å, 150 × 50 mm column, eluted with mobile phases: A) water (0.1% TFA), B) acetonitrile at a flow rate of 50 mL / min. [Table 1-1] [Table 1-2] [Table 2-1] [Table 2-2] [Table 3-1] [Table 3-2] [Table 4]

[0326] Example 1A: Synthesis of compound (10). Scheme 5. Synthesis of compound (10). [ka] 3-(Methoxycarbonyl)-4-phenylbut-3-enoic acid (1) [ka] A solution of benzaldehyde (100 g, 942 mmol) and dimethyl succinate (206 g, 1.41 mol) in tert-butanol (500 mL) was added over 1 h to a refluxing solution of potassium tert-butoxide (158 g, 1.41 mol) in tert-butanol (1.5 L). The mixture was then stirred for an additional 30 min before being allowed to cool to room temperature. After concentration in vacuo, the resulting residue was diluted with water (500 mL) and extracted with ethyl acetate (500 mL). The aqueous phase was then acidified to pH = 4-5 with aqueous hydrochloric acid (6 M) and then extracted with ethyl acetate (1 L). The combined organic extracts were dried over sodium sulfate, filtered, and concentrated in vacuo to give the title compound (300 g, impure) as a yellow oil, which was used in the next step without further purification. MS (ES+): m / z = 221 (M+H). + ;LCMS(Method F):t R =3.23 minutes. Methyl 4-hydroxy-2-naphthoate (2) [ka] A solution of 3-(methoxycarbonyl)-4-phenylbut-3-enoic acid (1) (300 g) and trifluoroacetic anhydride (99.3 mL, 714 mmol) in tetrahydrofuran (1.5 L) was stirred at 70 °C for 5 h, after which TLC confirmed consumption of the starting material. The reaction mixture was then concentrated in vacuo, adjusted to pH = 8-9 with aqueous sodium hydroxide (1 M), and extracted with ethyl acetate (1 L). The organic phase was dried over sodium sulfate and concentrated in vacuo. Recrystallization from ethyl acetate / petroleum spirit at 40-60 °C (10%) gave the title compound as a yellow solid (100 g, 53%). MS (ES+): m / z = 202 (M+H). + ;LCMS(Method F):t R =3.55 minutes. 4-(benzyloxy)-2-naphthoic acid methyl ester (3) [ka] A solution of methyl 4-hydroxy-2-naphthoate (2) (200 g, 990 mmol), benzyl bromide (203 g, 1.19 mol), and cesium carbonate (386 g, 1.19 mol) in N,N-dimethylformamide (800 mL) was stirred at 90° C. for 16 hours, after which TLC confirmed consumption of the starting material. The mixture was diluted with ethyl acetate (1.5 L), washed with water (1 L × 2) and then brine (500 mL), dried over sodium sulfate, and concentrated in vacuo to give the title compound (250 g, 86%) as a white solid, which was used in the next step without further purification.

[0327] 4-(benzyloxy)-2-naphthoic acid (4) [ka] A solution of methyl 4-(benzyloxy)-2-naphthoate (3) (250 g, 856 mmol) in toluene (500 mL) was charged with aqueous sodium hydroxide (12 M, 300 mL) and heated to 100 °C for 16 h, after which TLC confirmed consumption of the starting material. The organic phase was separated and concentrated in vacuo. The residue was then taken up in ethyl acetate (1.5 L) and acidified to pH = 2 with aqueous hydrochloric acid (6 M). The organic phase was separated, dried over sodium sulfate, and concentrated in vacuo. Recrystallization from ethyl acetate / petroleum spirit at 40-60 °C (10%) gave the title compound as a white solid (90 g, 32%). MS (ES+): m / z = 279 (M+H). + ;LCMS(Method F):t R =4.09 minutes.

[0328] tert-Butyl (4-(benzyloxy)naphthalen-2-yl)carbamate (5) [ka] A solution of 4-(benzyloxy)-2-naphthoic acid (4) (50.0 g, 180 mmol), diphenylphosphoryl azide (41.5 mL, 234 mmol), and triethylamine (28.9 mL, 270 mmol) in toluene (300 mL) was stirred at room temperature for 1 hour, after which TLC showed consumption of the starting material. tert-Butanol (200 mL) was added, and the resulting mixture was stirred at 90 °C for 17 hours. It was then diluted with ethyl acetate (1.5 L) and water (500 mL). The organic phase was separated, dried over sodium sulfate, filtered, and concentrated in vacuo. Recrystallization from ethyl acetate / petroleum spirit at 40-60 °C (10%) gave the title compound as a pink solid (35 g, 56%). MS (ES+): m / z = 350 (M+H). + ;LCMS(Method F):t R =4.67 minutes.

[0329] tert-Butyl (4-(benzyloxy)-1-iodonaphthalen-2-yl)carbamate (6) [ka] A mixture of tert-butyl (4-(benzyloxy)naphthalen-2-yl)carbamate (5) (55.0 g, 157 mmol), iodic acid (5.50 g, 31.5 mmol), and iodine (16.0 g, 63 mmol) in methanol (400 mL) and water (100 mL) was stirred at 80° C. for 5 hours, after which TLC indicated consumption of the starting material. The mixture was diluted with water (1.0 L) and filtered. The resulting cake was washed with methanol (200 mL) and concentrated in vacuo to give the title compound (72 g, 96%) as a brown solid. MS (ES+): m / z=476 (M+H). + ;LCMS(Method E):t R =4.91 minutes.

[0330] tert-Buty®)-(4-(benzyloxy)-1-iodonaphthalen-2-yl)(oxiran-2-ylmethyl)carbamate (7) [ka] A solution of tert-butyl (4-(benzyloxy)-1-iodonaphthalen-2-yl)carbamate (6) (52 g, 109 mmol) in N,N-dimethylformamide (500 mL) was charged with sodium hydride (60% dispersion in mineral oil, 17 g, 425 mmol) and stirred at room temperature for 30 minutes. After that, (S)-oxiran-2-ylmethyl 3-nitrobenzenesulfonate (51 g, 197 mmol) was added and the resulting mixture was stirred for an additional 3 hours. TLC confirmed the consumption of the starting material. The reaction mixture was carefully poured into ice-water (500 mL) and extracted with ethyl acetate (1.0 L). The organic phase was separated, washed with water (500 mL) and brine (300 mL), then dried over sodium sulfate and concentrated in vacuo to give the title compound (55 g, 95%) as a white solid. 1 H NMR(400MHz,CDCl3)δ 8.33-8.32(m,1H),8.41-8.20(m,1H),7.59-7.48(m,4H),7.45-7.33(m,3H),6.94-6.83(m,1H),5.28(s,2H),4.15-4.09 (m,1H),3.50-3.42(m,1H),3.14-3.13(m,1H),2.82-2.60(m,1H),2.41(ddd,J=12.4,4.8,2.8Hz,1H),1.33-1.31(m,9H).

[0331] (S)-tert-Butyl 5-(benzyloxy)-1-(hydroxymethyl)-1,2-dihydro-3H-benzo[e]indole-3-carboxylate (8) [ka] Zinc chloride (1 M in tetrahydrofuran, 28 mL) was diluted in anhydrous tetrahydrofuran (40 mL) and cooled to 0° C. under an inert argon atmosphere. A solution of methyllithium (1.6 M in diethyl ether, 70.6 mL) was then added dropwise to the cooled mixture and stirred for 30 minutes before further cooling to −78° C. (Trimethylsilyl)isothiocyanate (4 mL, 28.2 mmol) was added dropwise to the reaction mixture at −78° C., which was then warmed to 0° C. for 30 minutes and then cooled again to −78° C. A solution of tert-buI(R)-(4-(benzyloxy)-1-iodonaphthalen-2-yl)(oxiran-2-ylmethyl)carbamate (7) (10 g, 18.8 mmol) in tetrahydrofuran (20 mL) was added dropwise to the reaction mixture at −78° C. for 30 min, then warmed to 0° C. for 1 h and then to room temperature for 30 min. After quenching with saturated aqueous ammonium chloride, the mixture was extracted with dichloromethane (500 mL × 3), and the combined organics were washed with brine (100 mL), dried over sodium sulfate, and concentrated in vacuo to give the title compound (10 g, impure), which was used in the next step without further purification. 1 H NMR(400MHz,CDCl3)δ 8.29(d,J=8.4Hz,1H),7.90(s,1H),7.71(d,J=8.2Hz,1H),7.55(d,J=6.8Hz,2H),7.51-7.40(m,3H),7.36-7.32(m,2H),5.27(s,2H),4. 22(d,J=11.4Hz,1H),4.13(t,J=10.0Hz,1H),4.01-3.95(m,1H),3.85(bs,1H),3.81-3.73(m,1H),1.60(s,9H);MS(ES+):m / z=406(M+H) + ;LCMS(Method F):t R =4.69 minutes.

[0332] (S)-tert-Butyl 5-(benzyloxy)-1-(chloromethyl)-1,2-dihydro-3H-benzo[e]indole-3-carboxylate (9) [ka] A solution of (S)-tert-butyl 5-(benzyloxy)-1-(hydroxymethyl)-1,2-dihydro-3H-benzo[e]indole-3-carboxylate (8) (10.0 g, 12.4 mmol), carbon tetrachloride (30 mL), and triphenylphosphine (3.90 g, 14.8 mmol) in dichloromethane (50 mL) was stirred at room temperature for 2 hours, after which time TLC indicated consumption of the starting material. The reaction mixture was then concentrated in vacuo. Purification by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirits at 40-60°C (10%), followed by recrystallization from dichloromethane / petroleum spirits at 40-60°C (90%), afforded the title compound as a white solid (1.47 g, 28%). [□]D 23 =-14.5 o (c0.470,CH2Cl2); 1 H NMR(400MHz,CDCl3)δ 8.29(d,J=8.4Hz,1H),7.86(s,1H),7.65(d,J=8.4Hz,1H),7.58-7.30(m,7H),5.27(s,2H),4.27-4.24(m,1 H),4.13(t,J=10.6Hz,1H),4.01-3.87(m,2H),3.44(t,J=10.4Hz,1H),1.61(s,9H);MS(ES+):m / z=424(M+H) + ;LCMS(Method D):t R =4.27 minutes.

[0333] (S)-1-(Chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) [ka] A solution of (S)-tert-butyl 5-(benzyloxy)-1-(chloromethyl)-1,2-dihydro-3H-benzo[e]indole-3-carboxylate (9) (100 mg, 0.236 mmol) in anhydrous dichloromethane (3 mL) was charged dropwise with boron trichloride (1 M solution in dichloromethane, 708 μL, 0.708 mmol) via syringe at room temperature and stirred under an inert argon atmosphere. The resulting orange solution was stirred for 5 minutes, then quenched by careful addition of methanol (5 mL) and then concentrated in vacuo. The residue was again charged with methanol (5 mL) and reconcentrated in vacuo. Diethyl ether (5 mL) was then added, and the residue was once again concentrated in vacuo. The residue was then subjected to high vacuum for 30 minutes to give the title compound (55 mg, impure) as a pale green crystalline solid (unstable), which was used directly in the next step (amide coupling) without further purification. MS(ES+): m / z=234(M+H) + ;LCMS(Method C):t R =2.62 minutes.

[0334] Example 1B: Synthesis of compound (16). Scheme 6. Synthesis of compound (16). [ka]

[0335] Methyl 4-bromo-1-methyl-1H-pyrrole-2-carboxylate (12) [ka] Sodium hydride (60% dispersion in mineral oil, 300 mg, 613 mmol) was diluted in N,N-dimethylformamide (10 mL) and stirred at 0°C. A solution of methyl 4-bromo-1H-pyrrole-2-carboxylate (11) (1.00 g, 4.90 mmol) was added over 10 minutes at 0°C. The resulting mixture was stirred at the same temperature for 30 minutes. Methyl iodide (1.04 g, 7.40 mmol) was then added dropwise to the reaction mixture. After stirring at room temperature for 18 hours, the mixture was poured into water and then extracted with ethyl acetate (20 mL × 2). The combined organic extracts were dried over magnesium sulfate and concentrated in vacuo. Purification by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit, 40–60°C (20%), afforded the title compound (0.96 g, 90%) as a white solid. 1 H NMR (400MHz, CDCl3) δ 6.90 (s, 1H), 6.77 (s, 1H), 3.90 (s, 3H), 3.81 (s, 3H).

[0336] Methyl 4-(4-((tert-butoxycarbonyl)amino)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (13) [ka] To a solution of methyl 4-bromo-1-methyl-1H-pyrrole-2-carboxylate (12) (1.12 g, 5.14 mmol) and tert-butyl (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)carbamate (1.97 g, 6.16 mmol) in toluene, water, and propan-2-ol (10 mL) was charged potassium carbonate (2.14 g, 15.5 mmol) and tetrakis(triphenylphosphine)palladium(0) (0.36 g, 0.31 mmol). The resulting mixture was then heated to 110° C. for 16 hours, after which it was cooled to room temperature and diluted with water (20 mL). The mixture was partitioned between ethyl acetate and brine, and the organic layer was then dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit, 40-60° C. (33%) gave the title compound as an off-white solid (0.87 g, 51%). 1 H NMR(400MHz,CDCl3)δ 7.42-7.37(m,2H),7.36-7.32(m,2H),7.16(d,J=2.0Hz,1H),7.02(d,J=2.0Hz,1H),6.56(s,1H),3.94(s,3H),3.83(s,3H),1.45(s,9H); 13 C NMR(100MHz,CDCl3)δ 161.7,136.5,129.4,127.1,125.9,125.5,123.6,119.0,115.6,114.6,60.4,51.1,36.9,28.3;MS(ES+):m / z=331(M+H) + ;LCMS(Method B):t R =4.22 minutes.

[0337] Methyl 4-(4-aminophenyl)-1-methyl-1H-pyrrole-2-carboxylate (14) [ka] A solution of methyl 4-(4-((tert-butoxycarbonyl)amino)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (13) (4.8 g, 14.5 mmol) in 1,4-dioxane (20 mL) was charged with HCl (4 M in 1,4-dioxane, 20 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 hours, after which time TLC showed the reaction to be complete. The reaction was then quenched with saturated aqueous sodium bicarbonate and extracted with ethyl acetate (20 mL × 2). The combined organic extracts were then dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit, 40–60° C. (25%), afforded the title compound (3.1 g, 92%) as a pale yellow solid. 1 H NMR(400MHz,DMSO-d6)δ 7.36(s,1H),7.21(d,J=8.0Hz,2H),7.03(s,1H),6.54(d,J=8.0Hz,2H),5.00(s,2H),3.85(s,3H),3.74(s,3H);MS(ES+):m / z=231(M+H) + ;LCMS(Method F):t R =2.14 minutes.

[0338] Methyl 4-(4-(4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (15) [ka] To a solution of methyl 4-(4-aminophenyl)-1-methyl-1H-pyrrole-2-carboxylate (14) (2.18 g, 9.47 mmol) and 4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxylic acid (2.73 g, 11.4 mmol) in N,N-dimethylformamide (20 mL) was charged 4-dimethylaminopyridine (2.89 g, 23.7 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (3.63 g, 18.9 mmol). The resulting mixture was stirred at room temperature for 16 hours, then quenched with saturated aqueous sodium bicarbonate and extracted with ethyl acetate (20 mL × 2). The combined organic extracts were then dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by flash column chromatography (silica) eluting with methanol / dichloromethane (2%) gave the title compound as a pale yellow solid (3.61 g, 84%). 1 H NMR(400MHz,CDCl3)δ 8.01(s,1H),7.71(s,1H),7.54-7.49(m,2H),7.44-7.40(m,2H),7.17(d,J=2.0Hz,1H),7.03(d ,J=2.0Hz,1H),6.85(s,1H),6.63(s,1H),3.94(s,3H),3.88(s,3H),3.83(s,3H),1.50(s,9H); 13 C NMR(100MHz,CDCl3)δ 161.7,159.5,136.0,130.4,126.0,125.5,123.5,123.4,121.8,120.3,11 8.6,114.6,110.0,103.7,51.1,36.9,36.7,28.4;MS(ES+):m / z=453(M+H) + ;LCMS(Method F):t R =3.85 minutes.

[0339] Methyl 4-(4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (16) [ka] To a solution of methyl 4-(4-(4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (15) (2.42 g, 5.35 mmol) in 1,4-dioxane (20 mL) was charged HCl (4 M in 1,4-dioxane, 20 mL) at room temperature. The resulting mixture was stirred at room temperature for 3 hours, after which TLC revealed consumption of the starting material. The reaction was then quenched with saturated aqueous sodium bicarbonate and extracted with ethyl acetate (20 mL × 2). The combined organic extracts were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was triturated with dichloromethane / diethyl ether (1:8) (30 mL) to give the title compound (1.71 g, 90%) as a pale yellow solid. 1 H NMR(400MHz,DMSO-d6)δ 9.52(s,1H),7.67(d,J=8.0Hz,2H),7.55(s,1H),7.49(d,J=8.0Hz,2H),7.19(s,1H),6. 46(s,1H),6.32(s,1H),3.88(br,5H),3.76(s,3H),3.73(s,3H);MS(ES+):m / z=353(M+H) + ;LCMS(Method F):t R =2.41 minutes.

[0340] Example 1C: Synthesis of compound (18). Scheme 7. Synthesis of compound (18). [ka]

[0341] 6-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-6-oxohexanoic acid (17) [ka] A solution of adipic acid (38 mg, 0.26 mmol) in dichloromethane (0.5 mL) was charged with triethylamine (151 μL, 1.08 mmol) and HATU (103 mg, 0.27 mmol). After stirring for 5 min, methyl 4-(4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (16) (100 mg, 0.26 mmol) was added. The resulting mixture was stirred for 18 h, then charged with water (10 mL) and extracted with ethyl acetate (20 mL × 2). The aqueous layer was then acidified to pH = 6 by careful addition of hydrochloric acid (1 M, aq.) and extracted with ethyl acetate (20 mL × 2). The combined organic extracts were dried over magnesium sulfate and concentrated in vacuo. The resulting residue was used in the next step without further purification. MS (ES+): m / z = 480 (M+H) + ;LCMS(Method A):t R =6.33 minutes.

[0342] (S)-4-(4-(4-(6-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-6-oxohexanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (18) [ka] A solution of 6-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-6-oxohexanoic acid (17) (34 mg, 0.071 mmol) in N,N-dimethylacetamide (1 mL) was charged with (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (32 mg, 0.12 mmol), followed by the addition of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (68 mg, 0.36 mmol), and the resulting mixture was stirred at room temperature for 16 hours. Then, additional N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (68 mg, 0.36 mmol) was added, and the mixture was heated to 35°C with stirring for 3 hours. The mixture was then diluted into ethyl acetate (20 mL), extracted with cold brine (20 mL x 2), dried over magnesium sulfate, and concentrated in vacuo. Purification by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit at 40-60 °C (0%-100%) afforded the title compound (4.1 mg, 8%) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ 10.32(s,1H),9.82(s,1H),9.77(s,1H),8.06(d,J=8.5Hz,1H),7.96(s,1H),7.76(d,J=8.3Hz,1H),7.66( d,J=8.5Hz,2H),7.54(s,1H),7.49(d,J=8.3Hz,2H),7.46(d,J=8.2Hz,1H),7.32-7.25(m,1H),7.20(s,1H) ,7.18(s,1H),6.94(s,1H),4.31(t,J=10.2Hz,1H),4.14(m,2H),3.96(m,1H),3.87(s,3H),3.81(s,3H),3 .79-3.76(m,1H),3.75(s,3H),2.60-2.51(m,2H),2.28(m,2H),1.71-1.58(m,4H);MS(ES+):m / z=697(M+H) + ;LCMS(Method A):t R =7.80 minutes.

[0343] Example 1D: Synthesis of compound (22). Scheme 8. Synthesis of compound (22). [ka]

[0344] Methyl 4-(4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate hydrochloride (19) [ka] To a solution of methyl 4-(4-(4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (15) (1.0 g, 2.2 mmol) in 1,4-dioxane (15 mL) was charged HCl (4 M in 1,4-dioxane, 15 mL) and stirred at room temperature for 6 h. Upon completion, the reaction mixture was concentrated in vacuo, then diethyl ether (15 mL) was added and the residue was re-concentrated in vacuo to afford the title compound as a light brown solid, which was used directly in the next step without further purification. 1 H NMR(400MHz,DMSO-d6)δ 10.11(br,2H),9.98(s,1H),7.69(d,J=8.0Hz,2H),7.58(s,1H),7.53(d,J=8.0Hz,2H),7 .21(s,1H),7.20(s,1H),7.11(s,1H),3.89(br,6H),3.76(s,3H);MS(ES+):m / z=353(M+H) + ;LCMS(Method A):t R =5.38 minutes.

[0345] Methyl 4-(4-(4-(5-(tert-butoxy)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (20) [ka] A solution of pentanedioic acid mono-tert-butyl ester (125 mg, 0.67 mmol) in dichloromethane (1 mL) was charged with triethylamine (378 μL, 2.56 mmol) and HATU (258 mg, 0.67 mmol) and stirred at room temperature for 30 minutes. Then, methyl 4-(4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate hydrochloride (19) (250 mg, 0.64 mmol) was added, and the reaction mixture was allowed to stir at room temperature for 16 hours. Upon completion, the mixture was concentrated in vacuo. Flash column chromatography (silica) eluting with ethyl acetate / petroleum ether (30% to 80%) afforded the title compound as an off-white solid (187 mg, 56%). MS (ES+): m / z = 523 (M+H). + ;LCMS(Method A):t R =7.62 minutes.

[0346] 5-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoic acid (21) [ka] Methyl 4-(4-(4-(5-(tert-butoxy)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (20) (170 mg, 0.33 mmol) was dissolved in HCl (1 M in 1,4-dioxane, 4 mL) and allowed to stir at room temperature for 16 hours. Upon completion, the reaction mixture was charged with saturated aqueous sodium bicarbonate (10 mL) and extracted with ethyl acetate (15 mL x 2). The combined organic layers were then dried over magnesium sulfate and concentrated in vacuo to give the title compound as a yellow solid, which was used in the next step without further purification. MS (ES+): m / z = 467 (M+H). + ;LCMS(Method A):t R =6.47 minutes.

[0347] (S)-4-(4-(4-(5-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (22) [ka] A solution of 5-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoic acid (21) (94 mg, 0.20 mmol) in N,N-dimethylacetamide (1 mL) was added to (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (54 mg, 0.20 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (153 mg, 0.80 mmol). The reaction mixture was then stirred at room temperature for 16 hours. Upon completion, the mixture was diluted into ethyl acetate, washed with cold brine (20 mL × 2), dried over magnesium sulfate, and concentrated in vacuo. Flash column chromatography (silica) eluting with ethyl acetate / petroleum ether (60% to 100%) gave the title compound as a white solid (24 mg, 18%). 1H NMR(400MHz,DMSO-d6)δ 10.34(br,1H),9.85(br,1H),9.78(br,1H),8.06(d,J=8.2Hz,1H),7.99(s,1H),7.76(d,J=8.6Hz,1H ),7.67(d,J=8.2Hz,2H),7.49(m,4H),7.30(t,J=8.0Hz,1H),7.21(s,1H),7.18(s,1H),6.95(s,1H),4 .33-4.28(m,1H),4.14-4.12(m,1H),3.98-3.96(m,1H),3.87(s,3H),3.81(s,3H),3.80-3.78(m,1H), 3.75(s,3H),2.61-2.51(m,2H),2.36(t,J=7.4Hz,2H),1.19(t,J=7.4Hz,2H);MS(ES+):m / z=682(M+H) + ;LCMS(Method A):t R =7.67 minutes.

[0348] Example 1E: Synthesis of compound (24). Scheme 9. Synthesis of compound (24). [ka] 4-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutanoic acid (23) [ka] A solution of succinic acid (42 mg, 0.38 mmol) in dichloromethane (1 mL) was charged with triethylamine (225 μL, 1.56 mmol) and HATU (148 mg, 0.67 mmol) and stirred at room temperature for 30 minutes. Then, methyl 4-(4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (16) (150 mg, 0.38 mmol) was added, and the reaction mixture was allowed to stir at room temperature for 16 hours. Upon completion, the mixture was concentrated in vacuo, and the residue was used directly in the next step without further purification. MS (ES+): m / z = 453 (M+H).+ ;LCMS(Method A):t R =6.45 minutes.

[0349] (S)-4-(4-(4-(4-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-4-oxobutanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (24) [ka] A solution of 4-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutanoic acid (23) (170 mg, 0.38 mmol) in N,N-dimethylacetamide (1 mL) was added to (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (101 mg, 0.38 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (288 mg, 1.50 mmol), and the resulting mixture was stirred at room temperature for 16 hours. The mixture was then diluted with ethyl acetate, washed with cold brine (20 mL × 2), dried over magnesium sulfate, and concentrated in vacuo. Flash column chromatography (silica) eluting with ethyl acetate / petroleum ether (70% to 100%) gave the title compound as a white solid (13 mg, 5%). 1H NMR(400MHz,DMSO-d6)δ 10.31(br,1H),9.93(br,1H),9.77(br,1H),8.06(d,J=8.2Hz,1H),7.94(s,1H),7.77(d,J=8.6Hz ,1H),7.67(d,J=8.6Hz,2H),7.53(s,1H),7.50-7.46(m,3H),7.30(t,J=7.6Hz,1H),7.19-7.18(m, 2H),6.97(s,1H),4.36(t,J=10.0Hz,1H),4.21-4.16(m,2H),4.01-3.98(m,1H),3.87(s,3H),3.8 1(s,3H),3,79(s,1H),3.75(s,3H),2.84-2.77(m,2H),2.63-2.60(m,2H);MS(ES+):m / z=669(M+H) + ;LCMS(Method A):t R =7.75 points.

[0350] Example 1F: Synthesis of compound (29). スキーム10. Synthesis of compound (29).

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[0351] 6-アミノイミダゾ[1,2-a]ピリジン-2-カルボンエチル(25)

change

[0352] Ethyl 6-(4-(methoxymethoxy)benzamido)imidazo[1,2-a]pyridine-2-carboxylate (26) [ka] A solution of 4-(methoxymethoxy)benzoic acid (216 mg, 1.18 mmol) in N,N-dimethylacetamide (1 mL) was charged with N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (272 mg, 1.42 mmol) and stirred at room temperature for 2 minutes. A pre-sonicated solution of ethyl 6-aminoimidazo[1,2-a]pyridine-2-carboxylate (25) (243 mg, 1.18 mmol) in N,N-dimethylacetamide (1.5 mL) was then added, and the resulting mixture was stirred at room temperature for 16 hours. After dilution in ethyl acetate (10 mL), the mixture was washed with cold brine (10 mL x 2), dried over magnesium sulfate, and concentrated in vacuo to give the title compound (284 mg, 65%) as a green oil, which was used in the next step without further purification. MS (ES+): m / z = 370 (M+H). + ;LCMS(Method A):t R =5.83 minutes.

[0353] 6-(4-(methoxymethoxy)benzamido)imidazo[1,2-a]pyridine-2-carboxylic acid (27) [ka] A solution of ethyl 6-(4-(methoxymethoxy)benzamido)imidazo[1,2-a]pyridine-2-carboxylate (26) (284 mg, 0.77 mmol) in tetrahydrofuran (2 mL) was charged with potassium hydroxide (1 M, aq., 2.3 mL) at room temperature and stirred for 1 h. The red / brown solution was then quenched by the careful addition of saturated aqueous citric acid until a pH of 6 was reached. The resulting yellow precipitate was filtered under reduced pressure and washed with ethyl acetate and water. A strong vacuum was then applied to the filtered solid for 30 min to give the title compound (235 mg, 90%) as a very pale yellow solid, which was used in the next step without further purification. MS (ES+): m / z = 342 (M+H). + ;LCMS(Method A):t R =4.88 minutes.

[0354] (S)-N-(2-(1-(chloromethyl)-5-hydroxy-2,3-dihydro-1H-benzo[e]indole-3-carbonyl)imidazoXXXorbilipyridin-6-yl)-4-(methoxymethoxy)benzamide (28) [ka] A slurry of 6-(4-(methoxymethoxy)benzamido)imidazo[1,2-a]pyridine-2-carboxylic acid (27) (80 mg, 0.23 mmol) in N,N-dimethylacetamide (1 mL) was charged with (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (66 mg, 0.24 mmol), followed by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (137 mg, 0.71 mmol), and the resulting mixture was stirred for 16 h. The mixture was then diluted with ethyl acetate (10 mL), washed with cold brine (10 mL x 2), dried over magnesium sulfate, and concentrated in vacuo. Purification was carried out by flash column chromatography (silica) eluting with ethyl acetate / petroleum ether (0% to 100%) to give the title compound (50 mg, 38%) as a very pale yellow solid. 1 H NMR (400 MHz, DMSO-d6)

[0355] δ 10.40(s,1H),10.29(s,1H),9.44-9.40(m,1H),8.63(s,1H),8.10(d,J=8.2Hz,1H),8.03(br,1H),7.98(d,J= 9.3Hz,2H),7.81(d,J=8.4Hz,1H),7.70(d,J=9.7Hz,1H),7.56(dd,J=9.8,2.0Hz,1H),7.53-7.47(m,1H),7.37 -7.31(m,1H),7.16(d,J=9.3Hz,2H),5.29(s,2H),4.95(d,J=11.4Hz,1H),4.75(dd,J=13.5,6.9Hz,1H),4.20- 4.14(m,1H),3.98(dd,J=11.1,3.0Hz,1H),3.78(dd,J=11.0,7.8Hz,1H),3.39(s,3H);MS(ES+):m / z=557(M+H) + ;LCMS(Method A):t R =6.70 minutes.

[0356] (S)-N-(2-(1-(chloromethyl)-5-hydroxy-2,3-dihydro-1H-benzo[e]indole-3-carbonyl)imidazoXXXorbilipyridin-6-yl)-4-hydroxybenzamide (29) [ka] (S)—N-(2-(1-(chloromethyl)-5-hydroxy-2,3-dihydro-1H-benzo[e]indole-3-carbonyl)imidazo[1,2-a]pyridin-6-yl)-4-(methoxymethoxy)benzamide (28) (45 mg, 0.081 mmol) was charged with HCl (4 M in 1,4-dioxane, 2 mL) and the resulting green slurry was stirred at room temperature for 10 minutes, at which point excess cold diethyl ether was added. After concentration in vacuo, the mustard-colored residue was purified by flash column chromatography (silica) eluting with methanol / ethyl acetate (0% to 55%) to give the title compound (5.47 mg, 13%) as a very pale yellow solid. 1H NMR(400MHz,DMSO-d6)δ 10.20(s,1H),9.42(s,1H),8.63(s,1H),8.10(d,J=8.4Hz,1H),8.03(br,1H),7.89(d,J=8.2H z,2H),7.81(d,J=8.1Hz,1H),7.69(d,J=9.7Hz,1H),7.57(dd,J=9.8,1.2Hz,1H),7.53-7.47(m ,1H),7.37-7.31(m,1H),6.89(d,J=8.1Hz,2H),4.94(d,J=11.3Hz,1H),4.74(t,J=10.5Hz,1H ),4.20-4.14(m,1H),4.01-3.94(m,1H),3.78(dd,J=11.2,7.9Hz,1H);MS(ES+):m / z=513(M+H) + ;LCMS(Method A):t R =6.17 minutes.

[0357] Example 1G: Synthesis of compound (37). Scheme 11. Synthesis of compound (37). [ka]

[0358] 4-(4-(4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylic acid (30) [ka] A solution of methyl 4-(4-(4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (15) (5.00 g, 11.0 mmol) in tetrahydrofuran (15 mL), methanol (5 mL), and water (5 mL) was charged with lithium hydroxide (1.30 g, 54.2 mmol) and stirred at 30° C. for 16 hours. The resulting mixture was then partially concentrated in vacuo (to remove the methanol and tetrahydrofuran) to give a viscous emulsion, which was then diluted in ethyl acetate (250 mL) and acetone (10 mL). After acidification to pH 5 with aqueous citric acid (1 M), the aqueous layer was separated and then extracted with ethyl acetate (250 mL), and the combined organic extracts were then dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting pale brown solid residue was then triturated in diethyl ether / hexane (1:1) with rapid stirring, then filtered under reduced pressure and dried under strong vacuum to give the title compound as a very pale yellow solid (3.3 g, 69%). 1 H NMR(400MHz,DMSO-d6)δ 12.23(s,1H),9.75(s,1H),9.11(s,1H),7.70(d,J=8.7Hz,2H),7.53-7.44(m,3H),7.13(d,J= 2.2Hz,1H),6.99-6.90(m,2H),3.87(s,3H),3.81(s,3H),1.46(s,9H);MS(ES+):m / z=439(M+H) + ;LCMS(Method A):t R =7.02 minutes.

[0359] tert-Butyl (5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamate (31) [ka] To a solution of 4-(4-(4-((tert-butoxycarbonyl)amino)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylic acid (30) (3.30 g, 7.53 mmol) in N,N-dimethylformamide (15 mL) was charged triethylamine (4.40 mL, 31.6 mmol) and HATU (3.00 g, 7.90 mmol), and the resulting mixture was stirred at room temperature for 10 minutes, after which 1,4-diaminobenzene (814 mg, 7.53 mmol) was added and stirred for 16 hours. The reaction mixture was then diluted into ethyl acetate (250 mL) and washed with cold brine (100 mL) and aqueous sodium bicarbonate (100 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated in vacuo. Trituration with diethyl ether / hexane (1:1) with rapid stirring, followed by vacuum filtration and drying under high vacuum, gave the title compound as a mustard-colored solid (4 g, quantitative). 1 H NMR(600MHz,DMSO-d6)δ 9.75(s,1H),9.47(s,1H),9.11(s,1H),7.70(d,J=8.6Hz,2H),7.47(d,J=8.6Hz,2H),7.38(d,J=1.8Hz,1H),7.33(d,J=9.0Hz,2H),7.2 9(d,J=1.8Hz,1H),6.93(br,2H),6.53(d,J=9.0Hz,2H),4.86(br,2H),3.89(s,3H),3.81(s,3H),1.46(s,9H);MS(ES+):m / z=529(M+H) + ;LCMS(Method A):t R =6.25 minutes.

[0360] tert-Butyl (5-((4-(5-((4-((allyloxy)carbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamate (32) [ka] A solution of tert-butyl (5-((4-(5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamate (31) (4.00 g, 7.57 mmol) in N,N-dimethylacetamide (15 mL) was cooled to 0 °C and charged with pyridine (1.40 mL, 17.4 mmol), followed by the dropwise addition of allyl chloroformate (885 μL, 8.32 mmol). After stirring at 0 °C for 30 min, the reaction mixture was precipitated from diethyl ether and then vacuum filtered. The resulting solid was taken up in hot methanol and then vacuum filtered again to give the title compound (3.46 g, 75%) as a fine yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.78(s,2H),9.66(s,1H),9.14(s,1H),7.73(d,J=8.6Hz,2H),7.65(d,J=8.9Hz ,2H),7.50(d,J=8.6Hz,2H),7.46-7.39(m,4H),6.96(s,2H),6.01(ddd,J=22.5 ,10.6,5.4Hz,1H),5.38(dd,J=17.2,1.6Hz,1H),5.26(d,J=10.5Hz,1H),4.63( d,J=5.4Hz,2H),3.92(s,3H),3.84(s,3H),1.49(s,9H);MS(ES+):m / z=613(M+H) + ;LCMS(Method A):t R =7.75 minutes

[0361] 5-((4-(5-((4-((allyloxy)carbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-aminium 2,2,2-trifluoroacetate (33) [ka] A solution of tert-butyl (5-((4-(5-((4-((allyloxy)carbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamate (32) (3.00 g, 4.90 mmol) in dichloromethane (10 mL) and trifluoroacetic acid (1.9 mL, 24.5 mmol) was stirred at room temperature for 16 hours. The reaction mixture was then precipitated from diethyl ether and concentrated in vacuo. Diethyl ether (200 mL) was then added, and the mixture was concentrated in vacuo again. A strong vacuum was then applied to the residue for 1 hour to give the title compound (3.07 g, quantitative) as a light brown solid. 1 H NMR(400MHz,DMSO-d6)δ 10.04(s,2H),9.98(s,1H),9.81(s,1H),9.66(s,1H),7.73(d,J=8.6Hz,2H),7.66 (d,J=8.9Hz,2H),7.53(d,J=8.9Hz,2H),7.47(s,1H),7.43(d,J=7.3Hz,3H),7.18( s,1H),7.12(s,1H),6.01(ddd,J=22.4,10.6,5.4Hz,1H),5.38(d,J=17.3Hz,1H),5 .26(d,J=10.5Hz,1H),4.63(d,J=5.3Hz,2H),3.92(s,6H);MS(ES+):m / z=513(M+H) + ;LCMS(Method A):t R =5.62 minutes.

[0362] tert-Butyl 5-((5-((4-(5-((4-((allyloxy)carbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoate (34) [ka] A solution of 5-(tert-butoxy)-5-oxopentanoic acid (922 mg, 4.90 mmol) in N,N-dimethylformamide (5 mL) was charged with triethylamine (2.9 mL, 20.6 mmol) and HATU (1.95 g, 5.14 mmol), and the resulting mixture was stirred at room temperature for 10 minutes. A solution of 5-((4-(5-((4-((allyloxy)carbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-aminium 2,2,2-trifluoroacetate (33) (3.07 g, 4.90 mmol) in N,N-dimethylformamide (5 mL) was then added, and the resulting mixture was stirred at room temperature for 16 hours. Precipitation was induced by the addition of saturated aqueous sodium bicarbonate solution. After vacuum filtration, the resulting solid was taken up in hot methanol and vacuum filtered again to give the title compound as a very pale yellow solid (2.26 g, 67%). 1 H NMR(600MHz,DMSO-d6)δ 9.81(s,1H),9.79(s,1H),9.76(s,1H),9.63(br,1H),7.71(d,J=8.6Hz,2H),7.63(d,J=8.9Hz,2H),7.4 9(d,J=8.6Hz,2H),7.45-7.35(m,4H),7.20(d,J=1.7Hz,1H),6.96(d,J=1.8Hz,1H),5.99(ddd,J=22.6,1 0.7,5.4Hz,1H),5.36(dd,J=17.2,1.6Hz,1H),5.24(dd,J=10.5,1.4Hz,1H),4.60(d,J=5.4Hz,2H),3.9 0(s,3H),3.83(s,3H),2.27(t,J=7.4Hz,2H),2.24(t,J=7.4Hz,2H),1.78(p,J=7.5Hz,2H),1.41(s,9H); 13C NMR(150MHz,DMSO-d6)δ 172.4,169.4,160.1,160.0,153.7,137.6,134.9,134.6,133.9,130.0,126.7,125.7,124.9,123.2,122.5,122.4, 121.0,120.9,119.3,118.0,110.7,105.3,80.0,65.0,36.9,36.6,35.0,34.6,28.3,21.3;MS(ES+):m / z=683(M+H) + ;LCMS(Method A):t R =7.70 minutes.

[0363] 5-((5-((4-(5-((4-((allyloxy)carbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoic acid (35) [ka] To a slurry of tert-butyl 5-((5-((4-(5-((4-((allyloxy)carbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoate (34) (2.26 g, 3.31 mmol) in dichloromethane (7 mL) was charged trifluoroacetic acid (3 mL) to give an amber / brown solution which was stirred at room temperature for 16 hours. Additional trifluoroacetic acid (3 mL) was then charged and the mixture was stirred for an additional 2 hours. Once the reaction was judged complete by TLC and LCMS, the mixture was cooled to 0° C., diethyl ether (excess) was added and the resulting precipitate was filtered under reduced pressure. The precipitate was dried under strong vacuum to give the title compound (1.68 g, 81%) as a light brown solid. MS(ES+): m / z=627(M+H) + ;LCMS(Method A):t R =6.75 minutes.

[0364] (S)-(4-(4-(4-(4-(5-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxamido)phenyl) allyl carbamate (36) [ka] A slurry containing 5-((5-((4-(5-((4-((allyloxy)carbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoic acid (35) (1.68 g, 2.68 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (2.05 g, 10.7 mmol) in N,N-dimethylacetamide (5 mL) was sonicated for 10 min and then added to (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (765 mg, 2.83 mmol), and the resulting mixture was stirred at room temperature for 16 h. Additional (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (765 mg, 2.83 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.05 g, 10.7 mmol) were added to the reaction mixture, which was then stirred at room temperature for an additional 16 hours. Addition of cold water (excess) caused precipitation. After vacuum filtration, the solid residue was washed with water and then dried under strong vacuum to give the title compound (impurity, 3.0 g) as a gray / green solid, which was used in the next step without further intermediate purification. 1H NMR(600MHz,DMSO-d6)δ 10.40(s,1H),9.98(s,1H),9.84(s,2H),9.65(s,1H),8.08(d,J=8.4Hz, 1H),8.01(s,1H),7.77(d,J=8.3Hz,1H),7.72(d,J=8.5Hz,2H),7.65(d, J=8.5Hz,2H),7.49(d,J=8.6Hz,3H),7.44(s,2H),7.40(d,J=8.5Hz,2H),7.33-7.29(m,1H),7.23(s,1H),7.00(s,1H),5.98(ddd,J=22.6,10.7, 5.4Hz,1H),5.36(dd,J=17.2,1.5Hz,1H),5.24(d,J=10.4Hz,1H),4.60(d,J=5.4Hz,2H),4.32(t,J=10.4Hz,1H),4.14(d,J=8.8Hz,2H),3.98(d, J=8.5Hz,1H),3.90(s,3H),3.83(s,3H),3.81-3.77(m,1H),2.39(dd,J=8.9,4.7Hz,3H),1.94(dt,J=13.4,12.1Hz,3H);MS(ES+):m / z=842(M+H) + ;LCMS(Method A):t R =7.82 minutes.

[0365] (S)-N-(4-aminophenyl)-4-(4-(4-(5-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamide)phenyl)-1-methyl-1H-pyrrole-2-carboxamide (37) [ka] To a solution of (S)-(4-(4-(4-(4-(5-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)allyl carbamate (36) (30 mg, 0.036 mmol) in dichloromethane (1 mL) was charged boron trichloride (1 M solution in dichloromethane, 400 μL) and the resulting mixture was aged at room temperature under argon for 4 days. The reaction was quenched by the addition of methanol (10 mL) and then concentrated in vacuo. Purification by flash column chromatography (silica) eluting with methanol / dichloromethane (12%) afforded the title compound (6 mg, 22%) as a grey solid. 1 H NMR(600MHz,DMSO-d6)δ 10.35(s,1H),9.88-9.81(m,1H),9.77(s,1H),9.48(s,1H),8.08(d,J=8.2Hz,1H),8.00(s,1H),7.78(d,J=8.4Hz ,1H),7.70(d,J=8.7Hz,2H),7.48(dd,J=11.4,4.4Hz,2H),7.39(d,J=1.8Hz,1H),7.35-7.31(m,2H),7.29(t,J=5 .3Hz,1H),7.24-7.19(m,2H),6.99-6.93(m,2H),6.55-6.50(m,2H),4.87(br,2H),4.32(t,J=10.4Hz,1H),4.15( d,J=11.3Hz,2H),4.02-3.95(m,1H),3.89(s,3H),3.83(s,3H),3.79(dd,J=11.0,7.9Hz,1H),2.42-2.25(m,6H); 13C NMR(150MHz,DMSO-d6)δ 179.2,173.5,169.8,169.4,160.0,145.3,137.5,130.1,128.7,127.7,127.2,126.7,126.0,125.1,124.8,123.6,123.2,123.1,123 .0,122.6,122.4,122.3,121.0,119.3,114.2,110.1,105.3,53.1,52.5,51.7,36.8,36.6,35.0,33.1,21.1;MS(ES+):m / z=758(M+H) + ;LCMS(Method A):t R =6.93 minutes.

[0366] Example 1H: Synthesis of compound (39). Scheme 12. Synthesis of compound (39). [ka] 2-(3-(2-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)phenyl)acetic acid (38) [ka] A solution of 2-2-(1,3-phenylene)diacetic acid (171 mg, 0.88 mmol) in N,N-dimethylacetamide (1 mL) was charged with methyl 4-(4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (16) (150 mg, 0.43 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (186 mg, 0.97 mmol) and stirred at room temperature for 16 hours. The reaction mixture was then diluted into ethyl acetate (100 mL) and washed with cold brine (2 x 50 mL). After drying over magnesium sulfate and concentration in vacuo, the residue was purified by flash column chromatography (silica) eluting with methanol / ethyl acetate (10%) to give the title compound (86 mg, 38%) as a yellow oil. MS(ES+): m / z=529(M+H) + ;LCMS(Method A):t R =6.88 minutes.

[0367] (S)-4-(4-(4-(2-(3-(2-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-2-oxoethyl)phenyl)acetamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (39) [ka] A solution of 2-(3-(2-((5-((4-(5-(methoxycarbonyl)-1-methyl-1H-pyrrol-3-yl)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)phenyl)acetic acid (38) (86 mg, 0.16 mmol) in N,N-dimethylacetamide (1 mL) was charged with (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (43 mg, 0.16 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (123 mg, 0.64 mmol) and stirred at room temperature for 16 h. The reaction mixture was then diluted into ethyl acetate (100 mL) and washed with cold brine (2 x 50 mL). After drying over magnesium sulfate and concentration in vacuo, the residue was purified by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit, 40-60°C (50% to 100%) to give the title compound as an off-white solid (12 mg, 10%). 1 H NMR(600MHz,DMSO-d6)δ 10.36(s,1H),10.09(s,1H),9.78(s,1H),8.08(d,J=8.3Hz,1H),7.96(s,1H),7.77(d,J=8.4Hz,1H),7.68(d,J=8.7Hz, 2H),7.55(d,J=1.8Hz,1H),7.50(d,J=8.7Hz,2H),7.48(d,J=7.5Hz,1H),7.34-7.29(m,2H),7.28(d,J=1.8Hz,1H),7.25 -7.21(m,1H),7.21-7.17(m,3H),6.97(d,J=1.8Hz,1H),4.35(t,J=9.9Hz,1H),4.28(dd,J=10.7,1.8Hz,1H),4.13(t,J= 8.3Hz,1H),3.98-3.92(m,1H),3.91-3.87(m,4H),3.81(s,3H),3.77(s,3H),3.73(dd,J=11.0,8.1Hz,1H),3.59(s,2H); 13C NMR (150 MHz, DMSO-d6) δ 169.5,167.9,162.8,161.3,160.0,154.8,142.4,137.8,136.8,135.5,1 30.4,129.4,128.7,128.1,127.7,127.5,125.1,123.6,123.3,123.2,12 3.1,122.7,122.5,122.2,120.8,119.3,114.5,114.3,105.4,100.3,53. 4,51.5,48.1,43.1,42.8,41.3,38.7,37.0,36.6;MS(ES+):m / z=744(M+H) + ;LCMS(Method A):t R =8.02 minutes.

[0368] Example 1I: Synthesis of compound (42). Scheme 13. Synthesis of compound (42). [ka] (S)-7-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-7-oxoheptanoic acid (41) [ka] A solution of pimelic acid (135 mg, 0.50 mmol) in N,N-dimethylacetamide (1 mL) was charged with (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (160 mg, 1.00 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (211 mg, 1.10 mmol), and the resulting mixture was stirred at room temperature for 16 h. After dilution in ethyl acetate (100 mL), washing with cold brine (2 × 50 mL) was performed. The organic phase was dried over magnesium sulfate and concentrated in vacuo. Flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit at 40-60 °C (50% to 100%) afforded the title compound as an off-white solid (138 mg, 59%). MS (ES+): m / z = 376 (M+H). + ;LCMS(Method A):tR =6.98 minutes.

[0369] (S)-4-(4-(4-(7-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-7-oxoheptanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (42) [ka] To a solution of (S)-7-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-7-oxoheptanoic acid (41) (133 mg, 0.29 mmol) in N,N-dimethylacetamide (1 mL) was charged 4-(4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate hydrochloride (19) (113 mg, 0.29 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (222 mg, 1.16 mmol), and the resulting mixture was stirred at room temperature for 16 h. After dilution in ethyl acetate (100 mL), the organic phase was washed with cold brine (2 × 50 mL), dried over magnesium sulfate, and concentrated in vacuo. Flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit, 40-60° C. (75% to 100%) gave the title compound as a white solid (13 mg, 6%). 1H NMR(600MHz,DMSO-d6)δ 10.32(s,1H),9.79(d,J=8.9Hz,2H),8.07(d,J=8.3Hz,1H),7.99(s,1H),7.77(d,J=8.4Hz,1H),7.68(d,J=8.7Hz,2H),7.55(d,J=1.9H z,1H),7.51(d,J=8.6Hz,2H),7.49-7.46(m,1H),7.31(t,J=7.7Hz,1H),7.20(dd,J=7.5,1.9Hz,2H),6.95(d,J=1.8Hz,1H),4.33(t,J=1 0.0Hz,1H),4.15(d,J=10.5Hz,2H),3.98(dd,J=10.4,2.2Hz,1H),3.89(s,3H),3.82(s,3H),3.81-3.77(m,1H),3.77(s,3H),2.59-2.5 2(m,1H),2.46(dd,J=16.1,7.8Hz,1H),2.27(t,J=7.4Hz,2H),1.63(dd,J=13.3,5.9Hz,4H),1.44-1.35(m,2H);MS(ES+):m / z=710(M+H) + ;LCMS(Method A):t R =7.88 minutes.

[0370] Example 1J: Synthesis of compound (43). Scheme 14. Synthesis of compound (43). [ka] (S)-4-(4-(4-(5-(5-hydroxy-1-methyl-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (43) [ka] To a solution of (S)-4-(4-(4-(5-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)-1-methyl-1H-pyrrole-2-carboxylate (22) (10 mg, 0.015 mmol) in tetrahydrofuran (1 mL) was charged ammonium formate (7.4 mg, 0.117 mmol) and palladium on charcoal (10 wt.%) (10 mg), and the resulting mixture was heated to 65° C. under argon for 3 h. After cooling to room temperature, the mixture was filtered through Celite, and the resulting cake was washed with ethyl acetate and water. The phases were separated, the organic layer was washed with brine, and concentrated in vacuo to give the title compound (7.7 mg, 81%) as a white solid. 1 H NMR(600MHz,DMSO-d6)δ 10.16(s,1H),9.85(s,1H),9.79(s,1H),8.07(d,J=8.3Hz,1H),7.99(s,1H),7.72(dd,J=8.6,4.9Hz,1H),7.70 -7.66(m,2H),7.56(t,J=6.0Hz,1H),7.51(d,J=8.7Hz,2H),7.45(dd,J=11.2,4.0Hz,1H),7.29(t,J=7.5Hz,1H ),7.22(d,J=1.7Hz,1H),7.19(t,J=3.6Hz,1H),6.96(d,J=1.8Hz,1H),4.32-4.25(m,1H),3.89(s,3H),3.83(s ,3H),3.77(br,4H),2.37(t,J=7.5Hz,2H),1.92(dt,J=14.6,7.3Hz,2H),1.29(t,J=6.9Hz,3H),1.24(br,3H); 13C NMR(150MHz,DMSO-d6)δ 171.1,169.8,161.3,160.1,153.7,140.7,137.8,130.2,129.4,127.5,127.2,125.1,123.5,123.3,123.2,122.9,122.7,122.6,12 2.2,120.9,120.3,119.3,114.3,105.3,100.4,56.9,51.5,37.0,36.6,35.3,34.9,33.3,30.9,21.9,21.0;MS(ES+):m / z=648(M+H) + ;LCMS(Method A):t R =7.60 minutes.

[0371] Example 1K: Synthesis of compound (51). Scheme 15. Synthesis of compound (51). [ka] Methyl 4-((allyloxy)carbonyl)amino)-1-methyl-1H-pyrrole-2-carboxylate (45) [ka] A solution of methyl 4-amino-1-methyl-1H-pyrrole-2-carboxylate hydrochloride (44) (2.9 g, 15.2 mmol) in dichloromethane (30 mL) was cooled to 0 °C and charged with pyridine (2.8 mL, 35.0 mmol), followed by allyl chloroformate (1.8 mL, 16.7 mmol). The resulting mixture was stirred for 35 min, then diluted with dichloromethane (30 mL) and washed with saturated aqueous copper sulfate (2 × 50 mL), brine (2 × 50 mL), and saturated aqueous sodium bicarbonate (50 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated in vacuo to give the title compound (3.4 g, 94%) as a yellow solid. 1H NMR(600MHz,DMSO-d6)δ 9.45(s,1H),7.09(s,1H),6.66(s,1H),6.08-5.85(m,1H),5.35-5.29(m,1H),5.21(d dd,J=10.5,2.9,1.4Hz,1H),4.57(dd,J=16.7,3.4Hz,2H),3.80(s,3H),3.71(s,3H); 13 C NMR(150MHz,DMSO-d6)δ 161.2,153.6,133.9,123.2,119.8,119.4,117.8,108.0,65.0,51.4,36.6;MS(ES+):m / z=239(M+H) + ;LCMS(Method A):t R =6.32 minutes.

[0372] 4-(((allyloxy)carbonyl)amino)-1-methyl-1H-pyrrole-2-carboxylic acid (46) [ka] To a solution of methyl 4-(((allyloxy)carbonyl)amino)-1-methyl-1H-pyrrole-2-carboxylate (45) (3.4 g, 14.3 mmol) in tetrahydrofuran (30 mL) was charged an aqueous solution of sodium hydroxide (1 M) (140 mL), and the resulting mixture was rapidly stirred at room temperature for 48 h. The reaction was monitored by LCMS and, upon completion, quenched by the addition of aqueous citric acid (1 M) until the pH was adjusted to 3-4. It was then extracted with ethyl acetate (2 × 100 mL). The combined organic extracts were then washed with brine (2 × 100 mL) and water (100 mL), dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting residue was washed with dichloromethane and filtered to give the title compound (1.97 g, 62%) as a yellow solid. 1H NMR(600MHz,DMSO-d6)δ 12.29(br,1H),9.40(s,1H),7.03(s,1H),6.61(s,1H),5.95(ddd,J=22.5,10.6,5.4Hz,1H),5 .32(dd,J=17.2,1.1Hz,1H),5.21(dt,J=11.8,1.3Hz,1H),4.55(d,J=5.1Hz,2H),3.78(s,3H); 13 C NMR(150MHz,DMSO-d6)δ 162.3,153.6,134.0,122.9,120.4,119.3,117.8,108.1,65.0,36.6;MS(ES+):m / z=225(M+H) + ;LCMS(Method A):t R =5.33 minutes.

[0373] (5-((4-((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamate allyl (47) [ka] To a solution of 4-(((allyloxy)carbonyl)amino)-1-methyl-1H-pyrrole-2-carboxylic acid (46) (1.97 g, 8.79 mmol) in N,N-dimethylacetamide (18 mL) was charged N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (5.05 g, 26.4 mmol) and tert-butyl (4-aminophenyl)carbamate (2.20 g, 10.6 mmol), and the resulting mixture was stirred at room temperature for 18 hours. It was then diluted in ethyl acetate (100 mL) and washed with brine (3×50 mL). The organic phase was then dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting residue was precipitated with dichloromethane and diethyl ether to give the title compound (3.26 g, 90%) as a very pale yellow solid. 1H NMR(600MHz,DMSO-d6)δ 9.68(s,1H),9.44(s,1H),9.22(s,1H),7.56(d,J=9.0Hz,2H),7.36(d,J=8.5Hz,2H),6.96(s,1H),6.88(s,1H),6.01 -5.92(m,1H),5.33(d,J=17.2Hz,1H),5.22(dd,J=10.5,1.4Hz,1H),4.57(d,J=5.2Hz,2H),3.81(s,3H),1.47(s,9H); 13 C NMR(150MHz,DMSO-d6)δ 160.0,153.8,153.3,135.3,134.2,134.1,123.5,122.4,121.1,118.7,118.1,117.8,105.0,79.3,65.0,36.6,28.6;MS(ES+):m / z=415(M+H) + ;LCMS(Method A):t R =7.17 minutes.

[0374] tert-Butyl (4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)carbamate (48) [ka] A slurry of allyl (5-((4-((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamate (47) (500 mg, 1.21 mmol) in dichloromethane (5 mL) was charged with tetrakis(triphenylphosphine)palladium(0) (14 mg, 0.012 mmol) and pyrrolidine (121 μL, 1.45 mmol), and the resulting mixture was then sonicated until an amber solution formed, which was stirred at room temperature for 45 min. After filtration through Celite and washing with dichloromethane, the mixture was concentrated in vacuo and then purified by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit at 40-60 °C (0% to 100%) to give the title compound (261 mg, 65%) as a light brown solid. 1H NMR(600MHz,DMSO-d6)δ 9.41(s,1H),9.20(s,1H),7.56-7.53(m,2H),7.34(d,J=8.5Hz,2H),6.43(d, J=2.1Hz,1H),6.30(d,J=2.1Hz,1H),3.72(s,3H),2.74(s,2H),1.47(s,9H); 13 C NMR(150MHz,DMSO-d6)δ 160.1,153.3,135.0,134.5,132.2,123.1,120.9,118.7,116.6,115.3,79.2,36.2,28.6;MS(ES+):m / z=331(M+H) + ;LCMS(Method A):t R =5.32 minutes

[0375] 5-((5-((4-((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoic acid (49) - Method 1 [ka] tert-Butyl (4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)carbamate (48) (244 mg, 0.74 mmol) was treated with glutaric anhydride (93 mg, 0.81 mmol), 4-dimethylaminopyridine (9 mg, 0.074 mmol), and pyridine (2.5 mL). The reaction was allowed to stand at room temperature for 1 hour and then concentrated in vacuo. The residue was then diluted in ethyl acetate (30 mL) and washed sequentially with saturated aqueous ammonium chloride (5 × 20 mL) and brine (2 × 20 mL), then dried over anhydrous magnesium sulfate and concentrated in vacuo to provide the title compound (180 mg, 55%) as an off-white solid. 1H NMR(600MHz,DMSO-d6)δ 12.04(s,1H),9.81(s,1H),9.70(s,1H),9.23(s,1H),7.57-7.55(m,2H),7.37(d,J=8.7Hz,2H),7.19(d, J=1.8Hz,1H),6.92(d,J=1.9Hz,1H),3.82(s,3H),2.30-2.23(m,4H),1.80(p,J=7.4Hz,2H),1.47(s,9H); 13 C NMR(150MHz,DMSO-d6)δ 174.6,169.5,160.0,153.3,135.3,134.2,124.4,123.2,122.5,121.1,1 19.1,105.1,79.3,36.6,35.1,33.5,28.6,21.2;MS(ES+):m / z=445(M+H) + ;LCMS(Method A):t R =6.28 minutes.

[0376] 5-((5-((4-((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoic acid (49) - Method 2 [ka] A solution of glutaric acid (1.07 g, 8.11 mmol) in dichloromethane (4 mL) was charged with triethylamine (2.3 mL, 16.2 mmol) and HATU (1.54 g, 4.06 mmol) and stirred for 5 minutes before the addition of tert-butyl (4-(4-amino-1-methyl-1H-pyrrole-2-carboxamido)phenyl)carbamate (48) (1.34 g, 4.06 mmol). The reaction mixture was then stirred at room temperature for 18 hours before being concentrated in vacuo. Purification by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit at 40-60 °C (0% to 100%), followed by methanol / ethyl acetate (0% to 100%) afforded the title compound as a brown solid, which was used in the next step without further purification. MS (ES+): m / z = 445 (M+H). + ;LCMS(Method A):t R=6.38 minutes.

[0377] (S)-(4-(4-(5-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)tert-butylcarbamate (50) [ka] A solution of 5-((5-((4-((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoic acid (49) (1.8 g, 4.05 mmol) in N,N-dimethylacetamide (8 mL) was charged with (S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-ol hydrochloride (10) (1.4 g, 5.19 mmol) followed by N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (3.96 g, 20.7 mmol), and the resulting green slurry was sonicated and then stirred at room temperature for 48 hours. The mixture was then diluted with ethyl acetate (100 mL) and methanol (10 mL), extracted with brine (50 mL × 3), dried over magnesium sulfate, and concentrated in vacuo. Purification by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit, 40-60° C. (0% to 100%) gave the title compound as a rust-brown solid (792 mg, 30% over two steps). 1H NMR (600 MHz, CDCl3) δ 9.85(s,1H),8.47(s,1H),8.24(d,J=8.3Hz,1H),8.17-8.09(m,2H),7.84(s,1H) ,7.57(d,J=8.2Hz,1H),7.44(d,J=8.2Hz,2H),7.30(d,J=8.1Hz,2H),7.09(s,1H) ,6.63(br,1H),4.23-4.11(m,2H),3.97-3.82(m,2H),3.81(s,3H),3.45-3.33(m ,1H),2.70-2.54(m,4H),2.51-2.44(m,2H),1.50(s,9H);MS(ES+):m / z=660(M+H) + ;LCMS(Method A):t R =7.55 minutes.

[0378] (S)-N-(4-aminophenyl)-4-(5-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamide (51) [ka] To a solution of tert-butyl (S)-(4-(4-(5-(1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamido)phenyl)carbamate (50) (10 mg, 0.015 mmol) in dichloromethane (1 mL) was charged trifluoroacetic acid (1 mL), and the resulting mixture was stirred at room temperature for 1 min. After precipitation by the addition of diethyl ether, the mixture was concentrated in vacuo and then purified by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit at 40-60 °C (0% to 100%), followed by methanol / ethyl acetate (0% to 100%) to give the title compound as a white solid (1.2 mg, 14%). 1H NMR(600MHz,MeOD)δ 8.55(s,1H),8.17(d,J=8.3Hz,1H),7.93(s,1H),7.73(d,J=8.4Hz,1H),7.49(t,J=7.6Hz,1H),7.34-7.3 1(m,1H),7.28-7.26(m,2H),7.16(s,1H),6.82(s,1H),6.73-6.71(m,2H),4.56(br,2H),4.33-4.30(m,2 H),4.14-4.09(m,1H),4.12(dd,J=18.8,8.5Hz,1H),3.84(s,3H),3.64(dd,J=11.2,8.7Hz,1H),2.75-2. 68(m,1H),2.65-2.58(m,1H),2.48(dd,J=9.4,5.1Hz,2H),2.12(t,J=7.0Hz,2H);MS(ES+):m / z=560(M+H) + ;LCMS(Method A):t R =5.98 minutes.

[0379] Example 1L: Synthesis of compound (56). Scheme 16. Synthesis of compound (56). [ka]

[0380] (S)-tert-Butyl 1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indole-3-carboxylate (52) [ka] A solution of (S)-tert-butyl 5-(benzyloxy)-1-(chloromethyl)-1,2-dihydro-3H-benzo[e]indole-3-carboxylate (9) (1.6 g, 3.78 mmol) in dichloromethane (15 mL) and methanol (15 mL) was charged with palladium on carbon (10 wt.%) (160 mg) and Pearlman's catalyst (160 mg). The resulting mixture was stirred under a hydrogen atmosphere (1 atm) at room temperature overnight. The mixture was then filtered and concentrated in vacuo. Purification by flash column chromatography (silica) eluting with ethyl acetate / petroleum spirit at 40-60 °C (3%) afforded the title compound (1.3 g, 96%) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ 10.34(s,1H),8.07(d,J=8.4Hz,1H),7.74(d,J=8.4Hz,1H),7.47(ddd,J=8.4,6.8,1.4Hz,1H),7 .31-7.24(m,1H),4.14-3.93(m,4H),3.76(d,J=9.2Hz,1H),1.54(s,9H);MS(ES+):m / z=334(M+H) + ;LCMS(Method F):t R =4.37 minutes.

[0381] (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (53) [ka] A solution of (S)-1-(chloromethyl)-5-hydroxy-1,2-dihydro-3H-benzo[e]indole-3-carboxylate (52) (100 mg, 0.300 mmol) in dichloromethane (5 mL) was charged with 2,3,4,6-tetra-O-acetyl-α-D-galactopyranosyl trichloroacetimidate (184 mg, 0.375 mmol) and 4 Å molecular sieves (700 mg), and the resulting mixture was stirred at room temperature for 1 h. After cooling to −20 °C, boron trifluoride diethyl etherate (170 mg, 1.20 mmol) was added, and the resulting mixture was stirred for 3 h. The reaction was quenched by the addition of water (50 mL), and then the mixture was extracted with ethyl acetate (100 mL × 2). The combined organic extracts were then washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by preparative thin layer chromatography (silica) eluting with ethyl acetate / petroleum spirit, 40-60° C. (50%) gave the title compound as a brown oil (80 mg, 85%). 1 H NMR(400MHz,CDCl3)δ 7.98(d,J=8.4Hz,1H),7.58(d,J=8.4Hz,1H),7.45(s,1H),7.23(s,1H),6. 65(s,1H),5.68(dd,J=10.6,8.0Hz,1H),5.49(d,J=3.6Hz,1H),5.12(d,J= 8.2Hz,2H),4.33-4.24(m,1H),4.16-4.09(m,3H),3.96(t,J=9.4Hz,1H),3 .90-3.77(m,3H),3.51(s,1H),2.08-2.00(m,12H);MS(ES+):m / z=564(M+H) + ;LCMS(Method F):t R =1.92 minutes.

[0382] (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((S)-3-(5-((5-((4-((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoyl)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (54) [ka] To a solution of (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((S)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (53) (50 mg, 0.089 mmol) in N,N-dimethylacetamide (3 mL) was charged 5-((5-((4-((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoic acid (49) (26 mg, 0.059 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (34 mg, 0.178 mmol), and the resulting mixture was stirred at room temperature for 5 h. After dilution in water (50 mL), the mixture was extracted with ethyl acetate (50 mL x 2), and the combined organic extracts were washed with water and brine, dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by preparative thin-layer chromatography (silica) eluting with methanol / dichloromethane (5%) gave the title compound as a brown oil (15 mg, 17%). 1H NMR(400MHz,DMSO-d6)δ 9.84(s,1H),9.70(s,1H),9.23(s,1H),8.32(s,1H),7.97-7.82(m,2H),7.69-7.51(m,3H),7.46-7.32(m ,3H),7.23-7.18(m,1H),6.96-6.91(m,1H),5.62-5.51(m,1H),5.48-5.37(m,3H),4.60-4.48(m,1H),4. 46-4.29(m,1H),4.28-3.97(m,5H),3.93-3.86(m,1H),3.82(s,4H),2.69-2.57(m,2H),2.42-2.32(m,2H) ),2.18(s,3H),2.08(s,3H),2.04-2.00(m,3H),1.99-1.89(m,5H),1.47(s,9H);MS(ES+):m / z=990(M+H) + ;LCMS(Method F):t R =2.03 minutes.

[0383] (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((S)-3-(5-((5-((4-aminophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoyl)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (55) [ka] To a solution of (2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-(((S)-3-(5-((5-((4-((tert-butoxycarbonyl)amino)phenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-5-oxopentanoyl)-1-(chloromethyl)-2,3-dihydro-1H-benzo[e]indol-5-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (54) (15 mg, 0.015 mmol) in dichloromethane (2 mL) was charged trifluoroacetic acid (0.5 mL) and the resulting mixture was stirred at room temperature for 3 hours before being concentrated in vacuo. The title compound was used in the next step without further purification. MS (ES+): m / z = 890 (M+H). + ;LCMS(Method F):t R =3.34 minutes

[0384] N-(4-aminophenyl)-4-(5-((S)-1-(chloromethyl)-5-(((2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-1,2-dihydro-3H-benzo[e]indol-3-yl)-5-oxopentanamido)-1-methyl-1H-pyrrole-2-carboxamide (56) [...

Claims

1. A compound of formula (I): D-Q-B-T (I) or a salt, solvate, or tautomer thereof, wherein: D is a source of alkylated DNA minor groove binding units; Q is a linker, B is a DNA-binding amide-containing chain; T is a terminal group; wherein D, B, Q and / or T comprise at least one carbohydrate substituent.

2. The sugar chain substituent is R Ｓ and preferably R Ｓ The compound of claim 1, wherein is glycosyl or O-glycosyl.

3. D comprises a G that is a G-alkylated DNA group of formula (II): 【Chemistry 1】 During the ceremony, The dashed lines indicate the optional presence of double bonds between one or more of C1 and C2, C2 and C3, and C3 and C4; The wavy line indicates the point of attachment to Q; m is 0 or 1; R １ , R ３ , and R ４ are independently H and R ２９ and R ２ H, L ２ -R ２８、 R ２９ , and -L Ｓ -R Ｓ or R １ and R ２ , R ２ and R ３ Or R ３ and R ４ together with the carbon atom to which they are attached form a 6-membered aryl, or one, two, or three independently selected optional R ２０ forming a 5- or 6-membered cyclic, heterocyclic or heteroaryl ring optionally substituted by a group, R ５ and R ６ is (i) R ５ H, OH and OC １－６ alkyl, and R ６ But, H, SO ３ H, -L Ｓ -R Ｓ , nitrogen protecting group, -L ２ -R ２８ and R Ａ or (ii) R ５ is oxo or H, and R ６ But H or C １－６ alkyl, or (iii) R ５ and R ６ are selected so that either R ７ and R ９ are independently H and R ２０ is selected from R ８ H, SR ２４ , S.C.H. ２ Ph.R. ２０ , L ２ -R ２８ , and -L Ｓ -R Ｓ is selected from R Ａ is (CH ２ ) ｊ -OH, (CH ２ ) ｊ -CO ２ R ２６ , C(=O)-O-(CH ２ ) ｋ -NR ２６ R ２７ , (CH ２ ) ｊ NR ２６ R ２７ , C(=O)-NH-(CH ２ ) ｊ -NR ２６ R ２７ and C(=O)-NH-(CH ２ ) ｋ -C(=NH)NR ２６ R ２７ is selected from L ２ is a bond or a linker moiety having 1 to 200 non-hydrogen atoms selected from C, N, P, O, S or halogen, and which may incorporate an ether, oxo, carboxamidyl, uretanyl, branched, cyclic, unsaturated, heterocyclyl, aryl, or heteroaryl moiety; R ２８ is an azide, alkyne, bisulfone, carbohydrazide, hydrazine, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, pyridopyridazine, semihydrazide, succinimidyl ester, sulfodichlorophenol ester, sulfonyl halide, sulfosuccinimidyl ester, 4-sulfotetrafluorophenyl ester, tetrafluorophenyl ester, thiazole, R Ａ , O-(CH ２ ) ｋ -NR ２６ R ２６ , N.H.N.H. ２ or a targeting agent selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; Each R ２９ are independently R ２０ , R ２１ , =CH ２ , =CH-(CH ２ ) ｓ -CH ３ , =CH-(CH ２ ) ｓ -R ２１ , = O, (CH ２ ) ｓ -OR ２１ , (CH ２ ) ｓ -CO ２ R ２１ , (CH ２ ) ｓ -NR ２１ R ２４ , O-(CH ２ ) ｔ -NR ２１ R ２４ ,NH-C(O)-R ２１ , O-(CH ２ ) ｔ -NH-C(O)-R ２１ , O-(CH ２ ) ｔ -C(O)-NH-R ２１ , (CH ２ ) ｓ -SO ２ R ２１ , O-SO ２ R ２１ , (CH ２ ) ｓ -C(O)R ２１ and (CH ２ ) ｓ -C(O)NR ２１ R ２４ is selected from Each R ２０ are independently F, Cl, Br, (CH ２ ) ｊ -OH, C １－６ Alkyl, OC １－６ Alkyl, OCH ２ Ph, (CH ２ ) ｊ -CO ２ R ２６ , O-(CH ２ ) ｋ -NR ２６ R ２７ , C(=O)-O-(CH ２ ) ｋ -NR ２６ R ２７ , C(═O)—NR ２６ R ２７ , (CH ２ ) ｊ -NR ２６ R ２７ , N.R. ２６ NH ２ , C(=O)-NH-(CH ２ ) ｊ -NR ２６ R ２７ , C(=O)-NH-C ６ H ４ - (CH ２ ) ｊ -R ２６ , C(=O)-NH-(CH ２ ) ｋ -C(=NH)NR ２６ R ２７ , -L ２ -R ２８ , S(O) ２ -(C １－６ alkyl), O—(CH ２ ) ｋ -O-(C １－６ alkyl), (CH ２ ) ｊ -S(O) ２ -NR ２６ R ２７ , C(=NH)-O-(C １－６ alkyl), (CH ２ ) ｋ -O-(C １－６ alkyl), CN, NCO, Cy, C(O)—NH—(CH ２ ) ｊ -Cy, C(O)-Cy, NH-C(O)-NR ２６ R ２７ is selected from 【Chemistry 2】 each j and s is independently selected from 0, 1, 2, 3, 4, 5, or 6; each k and t is independently selected from 1, 2, 3, 4, 5, or 6; Each R ２１ are independently H, C １－１２ Alkyl, C ５－６ Heterocyclyl, C ５－９ Heteroaryl, C ６－１５ Heteroarylalkyl, phenyl, and C ７－１２ and aralkyl groups, wherein the heterocyclyl, heteroaryl, heteroarylalkyl, phenyl, and aralkyl groups are selected from one, two, or three independently selected optional R ２０ may be substituted with a group, Each R ２４ , R ２６ , and R ２７ are independently H and C １－１２ alkyl, Each Cy is independently C ５－６ Heterocyclyl or C ５－６ heteroaryl groups, wherein the heterocyclyl or heteroaryl group is selected from one or two R ２０ may be substituted with a group, L Ｓ is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH ２ ) １－１２ -, polyethylene glycol chain -(OCH ２ CH ２ ) １－６ - and these chains are P, O, S, NH, C ５－９ which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; ５－９ The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L Ｓ teeth, 【Transformation 3】 and L Ｃ is an amino acid, an amino acid derivative, a peptide chain having 2 to 6 amino acids or amino acid derivatives, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH ２ ) １－１２ -, polyethylene glycol chain -(OCH ２ CH ２ ) １－８ -, and these chains contain one or more groups selected from one or more P, O, S and / or NH groups, and / or C ５－９ may be interrupted by heteroarylene and / or phenylene, and each C ５－９ the heteroarylene group and / or each phenylene group may be optionally substituted; R Ｓ The compound of claim 1 or claim 2, wherein is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

4. The compound of claim 3, wherein G is selected from the group of formulas G1 to G8 below. 【Chemistry 4-1】 【Chemistry 4-2】

5. D comprises A, which is an A-alkylated DNA group of formula (IIIa) or (IIIb): 【Transformation 5】 During the ceremony, Z １ is a leaving group, optionally a halide, triflate, or tosylate; X is C-R １７ , N, N-R １７ , S or O, and the dashed line to X indicates the optional presence of a double bond depending on the nature of X; R １７ is H, -L Ｓ -R Ｓ , or R ２０ and R ２０ are independently F, Cl, Br, (CH ２ ) ｊ -OH, C １－６ Alkyl, OC １－６ Alkyl, OCH ２ Ph, (CH ２ ) ｊ -CO ２ R ２６ , O-(CH ２ ) ｋ -NR ２６ R ２７ , C(=O)-O-(CH ２ ) ｋ -NR ２６ R ２７ , C(═O)—NR ２６ R ２７ , (CH ２ ) ｊ -NR ２６ R ２７ , N.R. ２６ NH ２ , C(=O)-NH-(CH ２ ) ｊ -NR ２６ R ２７ , C(=O)-NH-C ６ H ４ - (CH ２ ) ｊ -R ２６ , C(=O)-NH-(CH ２ ) ｋ -C(=NH)NR ２６ R ２７ , -L ２ -R ２８ , S(O) ２ -(C １－６ alkyl), O—(CH ２ ) ｋ -O-(C １－６ alkyl), (CH ２ ) ｊ -S(O) ２ -NR ２６ R ２７ , C(=NH)-O-(C １－６ alkyl), (CH ２ ) ｋ -O-(C １－６ alkyl), CN, NCO, Cy, C(O)—NH—(CH ２ ) ｊ -Cy, C(O)-Cy, NH-C(O)-NR ２６ R ２７ is selected from 【Transformation 6】 L ２ is a bond or a linker moiety having 1 to 200 non-hydrogen atoms selected from C, N, P, O, S or halogen, and which may incorporate an ether, oxo, carboxamidyl, uretanyl, branched, cyclic, unsaturated, heterocyclyl, aryl, or heteroaryl moiety; R ２８ is an azide, alkyne, bisulfone, carbohydrazide, hydrazine, hydroxylamine, iodoacetamide, isothiocyanate, maleimide, phosphine, pyridopyridazine, semihydrazide, succinimidyl ester, sulfodichlorophenol ester, sulfonyl halide, sulfosuccinimidyl ester, 4-sulfotetrafluorophenyl ester, tetrafluorophenyl ester, thiazole, R Ａ , O-(CH ２ ) ｋ -NR ２６ R ２６ , N.H.N.H. ２ or a targeting agent selected from a protein, a portion of a protein, a peptide, a nucleic acid, or an antibody; R Ａ is (CH ２ ) ｊ -OH, (CH ２ ) ｊ -CO ２ R ２６ , C(=O)-O-(CH ２ ) ｋ -NR ２６ R ２７ , (CH ２ ) ｊ NR ２６ R ２７ , C(=O)-NH-(CH ２ ) ｊ -NR ２６ R ２７ and C(=O)-NH-(CH ２ ) ｋ -C(=NH)NR ２６ R ２７ is selected from Each R ２６ and R ２７ are independently H and C １－１２ alkyl, Each Cy is independently C ５－６ Heterocyclyl or C ５－６ heteroaryl groups, wherein the heterocyclyl or heteroaryl group is selected from one or two R ２０ may be substituted with a group, each j is independently selected from 0, 1, 2, 3, 4, 5, or 6; each k is independently selected from 1, 2, 3, 4, 5, or 6; z is r 1; R''' is OH or -L Ｓ -R Ｓ and L Ｓ is a bond, an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH ２ ) １－１２ -, polyethylene glycol chain -(OCH ２ CH ２ ) １－６ - and these chains are P, O, S, NH, C ５－９ which may be interrupted by or incorporate one or more of heteroarylene, phenylene, heterocyclyl, cycloalkyl, ether, oxo, carboxamidyl, and / or uretanyl moieties; ５－９ The heteroarylene, phenylene, heterocyclyl, and / or cycloalkyl moieties may be substituted and optionally include L Ｓ teeth, 【Transformation 7】 and R Ｓ The compound of claim 1 or claim 2, wherein is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

6. R １ , R ３ , R ７ , and R ９ 5. The compound of claim 3 or claim 4, wherein each of is H.

7. 5. The compound of claim 3 or claim 4, wherein the compound is a compound of formula IV: 【Transformation 8】

8. Q is X １ -L-X ２ Including, During the ceremony, X １ is O, S, NR １３ , C.R. １３ R １４ , C.R. １３ R １４ O, C(=O), C(=O)NR １３ , N.R. １３ selected from C(═O), O—C(O), and C(O)—O, or absent; L is an amino acid, a peptide chain having 2 to 6 amino acids, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH ２ ) １－１２ -, polyethylene glycol chain - (OCH ２ CH ２ ) １－６ -, these chains being selected from one or more P, O, S and / or NH groups, and / or C ５－９ may be interrupted by heteroarylene and / or phenylene, and each C ５－９ the heteroarylene group and / or each phenylene group may be optionally substituted; X ２ is O, S, NR １５ , C.R. １５ R １６ , C.R. １５ R １６ O, C(=O), C(=O)NR １５ , N.R. １５ selected from C(═O), O—C(O), and C(O)—O, or absent; R １３ , R １４、 R １５ , and R １６ are independently H and C １－６ 8. The compound of claim 1, wherein the aryl group is selected from alkyl.

9. B is (A) ｑ Including, During the ceremony, q is selected from 0, 1, 2, 3, 4, 5, and 6; A is selected from: 【Chemistry 9】 For each A group, Y ３ and Y ４ one of which is independently N-R ３０ , S, and O; Y ３ and Y ４ The other of these is CH, and Y ５ are independently, CR ３０ , N, S, and COH; For each A2 group, Y ６ and Y ７ is independently selected from N and CH; ６ and Y ７ The other one is CR ３０ and Each R ３０ are independently H, C １－６ Alkyl, L ２ -R ２８ , and R Ｓ 9. The compound according to any one of claims 1 to 8, selected from:

10. T comprises a group of the formula: 【Chemistry 10】 During the ceremony, p is 0 or 1; R Ｔ Is, -L ２ -R ２８ , phenyl, and C ５－９ heteroaryl, selected from phenyl and C ５－９ The heteroaryl group optionally includes OH, C １－６ Alkyl, OC １－６ Alkyl, -L ２ -R ２８ , (CH ２ ) ｊ -CO ２ R １１ , O-(CH ２ ) ｋ -NR １１ R １２ , (CH ２ ) ｊ -NR １１ R １２ , C(=O)-NH-(CH ２ ) ｋ -NR １１ R １２ , C(=O)-NH-R ２４ , and C(═O)—NH—(CH ２ ) ｋ -C(=NH)NR １１ R １２ and optionally, but optionally substituted with up to three optional substituents selected from ５－９ heteroaryl is other than indolyl; R １９ is H, C １－６ Alkyl, L ２ -R ２８ , R Ｓ , and (CH ２ ) ｔ -NR ２０ R ２１ is selected from Y １ and Y ２ are independently N or CR ３１ and Y １ and Y ２ At least one of the following is CR ３１ and Each R ３１ are independently H, C １－６ Alkyl, L ２ -R ２８ , and R Ｓ is selected from R １１ , R １２ , and R ２４ are independently H, -L ２ -R ２８ , and C １－６ 10. The compound of any one of claims 1 to 9, wherein the aryl group is selected from alkyl.

11. The compound is selected from compounds of the following formula: 【Chemistry 11-1】 【Chemistry 11-2】 【Chemistry 11-3】 【Chemistry 11-4】 【Chemistry 11-5】 【Chemistry 11-6】 【Chemistry 11-7】 In the formula, R Ｓ Compounds of formula (I) and salts and solvates thereof according to any one of claims 1 to 10, wherein is a monovalent saccharide substituent, preferably glycosyl or O-glycosyl.

12. The compound has at least one L ２ -R ２８ 12. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, comprising a group.

13. D, T, Q and / or B are L ２ -R ２８ 13. The compound of claim 12, or a pharmaceutically acceptable salt thereof, substituted with a group.

14. L ２ teeth, 【Chemistry 12】 is selected from In the formula, X ＡＡ is an amino acid sequence, and -K ２ is -[CH ２ CH ２ O- ０－５０ - or -[CH ２ ] ０－１２ 14. The compound of claim 12 or 13, wherein: -, or a pharmaceutically acceptable salt thereof.

15. L Ｓ is the following: 【Chemistry 13】 In the ceremony, L Ｃ is an amino acid, an amino acid derivative, a peptide chain having 2 to 6 amino acids or amino acid derivatives, an alkylene chain containing 1 to 12 carbon atoms which may contain one or more carbon-carbon double or triple bonds, a paraformaldehyde chain -(OCH ２ ) １－１２ -, polyethylene glycol chain -(OCH ２ CH ２ ) １－８ -, and these chains contain one or more groups selected from one or more P, O, S and / or NH groups and / or C ５－９ may be interrupted by heteroarylene and / or phenylene, and each C ５－９ 15. The compound of any one of claims 3 to 14, or a pharmaceutically acceptable salt thereof, wherein the heteroarylene group and / or each phenylene group is optionally substituted.

16. L Ｃ teeth, 【Chemistry 14】 at will 【Chemistry 15】 16. The compound of claim 15, comprising:

17. L Ｃ is a polyethylene glycol chain -(OCH ２ CH ２ ) １－８ -, optionally -(OCH ２ CH ２ ) ８ The compound according to claim 15 or 16, comprising:

18. R ２８ -L Ｃ 18. The compound of any one of claims 15 to 17, wherein: 【Chemistry 16】

19. X ＡＡ 19. The compound of any one of claims 12 to 18, or a pharmaceutically acceptable salt thereof, wherein is L-valyl-L-alanine.

20. R ２８ is the following maleimide: 【Chemistry 17】 20. The compound of any one of claims 12 to 19, or a pharmaceutically acceptable salt thereof, optionally linked to a targeting agent.

21. L ２ -R ２８ teeth, 【Chemistry 18-1】 【Chemistry 18-2】 Contains or 【Chemistry 19】 Contains or 【Chemistry 20】 Contains, or 【Chemistry 21】 Including, 21. The compound of any one of claims 12 to 20, or a pharmaceutically acceptable salt thereof, optionally linked to a targeting agent.

22. 20. A compound of formula (I) as claimed in any one of claims 21 to 17, and salts and solvates thereof, linked either directly or indirectly to a targeting agent to provide the conjugate of interest.

23. The compound has at least one L ２ -R ２８ group, and the targeting agent comprises the L ２ -R ２８ 23. The compound of formula (I) according to claim 22, and salts and solvates thereof, linked to said compound via a group.

24. 24. The compound of any one of claims 20 to 23, wherein the targeting agent comprises an antibody, an antibody fragment, a hormone, or a hormone fragment.

25. 25. A compound of formula (I) as defined in any one of claims 1 to 24, and salts and solvates thereof, for use as a pharmaceutical.

26. 25. Compounds of formula (I) as defined in any one of claims 1 to 24, and salts and solvates thereof, for use in the treatment of a proliferative disease.

27. 27. The compound of formula (I) as claimed in claim 26, and salts and solvates thereof, for use in the treatment of a proliferative disease, wherein the proliferative disease is selected from bladder cancer, osteosarcoma, intestinal cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, esophageal cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, retinoblastoma, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, and uterine cancer.

28. 25. A pharmaceutical composition comprising a compound of formula (I) as defined in any one of claims 1 to 24, and salts and solvates thereof, and a pharmaceutically acceptable excipient, carrier or diluent.

29. 30. Use of compounds of formula (I) as defined in any one of claims 1 to 28, and salts and solvates thereof, in the manufacture of a medicament for the treatment of a proliferative disease.

30. A method for treating a patient suffering from a proliferative disease, comprising administering to the patient a therapeutically effective amount of a compound of any one of claims 1 to 24 or a pharmaceutical composition of claim 28.

31. 31. The method of claim 30, wherein the proliferative disease is selected from bladder cancer, osteosarcoma, intestinal cancer, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, esophageal cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, retinoblastoma, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, and uterine cancer.

32. 25. An antibody-drug conjugate comprising a compound of formula (I) according to any one of claims 1 to 24, and salts and solvates thereof.

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