MCL-1 inhibitor antibody-drug conjugates and methods of use

Abstract

Anti-CD74 antibody-drug conjugates are disclosed. The anti-CD74 antibody-drug conjugates comprise an Mcl-1 inhibitor drug moiety and an anti-CD74 antibody or antigen-binding fragment thereof that binds an antigen target, e.g., an antigen expressed on tumor or other cancer cells. The present disclosure further relates to methods and compositions for use in treating cancer by administering the antibody-drug conjugates provided herein. Linker-drug conjugates comprising an Mcl-1 inhibitor drug moiety and methods for preparing the same are also disclosed.

Description

[Technical Field]

[0001] Related Applications This application claims benefit of and priority under 35 U.S.C. § 119(e) to the filing date of U.S. Provisional Patent Application No. 62 / 850,098, filed May 20, 2019, the entire contents of which are incorporated herein by reference.

[0002] The present disclosure relates to antibody-drug conjugates (ADCs) comprising an Mcl-1 inhibitor and an anti-CD74 antibody or antigen-binding fragment thereof that binds an antigen target, e.g., an antigen expressed on tumor or other cancer cells. The present disclosure further relates to methods and compositions useful in the treatment and / or diagnosis of cancers that express the target antigen CD74 and / or that can be applied to treatment by modulating Mcl-1 expression and / or activity, and methods for preparing the compositions. Linker-drug conjugates comprising an Mcl-1 inhibitor drug moiety and methods for preparing the same are also disclosed. [Background technology]

[0003] Apoptosis, or programmed cell death, is a physiological process crucial for embryonic development and the maintenance of tissue homeostasis. Apoptotic cell death is generally accompanied by morphological changes such as nuclear condensation and DNA fragmentation, as well as biochemical changes such as the activation of caspases, which can damage key structural components of the cell. The regulation of apoptosis is complex and typically involves the activation or suppression of several intracellular signaling pathways (Cory et al. (2002) Nature Review Cancer 2:647-656).

[0004] Deregulation of apoptosis is associated with certain pathological conditions. For example, increased apoptosis is associated with neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease, and ischemia. Conversely, defects in apoptosis may play a role in the development of cancer, as well as chemotherapy resistance, autoimmune diseases, inflammatory diseases, and viral infections. Lack of apoptosis is one of the phenotypic characteristics of cancer (Hanahan et al. (2000) Cell 100:57-70). Anti-apoptotic proteins of the Bcl-2 family are associated with many types of cancer, such as colon cancer, breast cancer, small cell lung cancer, non-small cell lung cancer, bladder cancer, ovarian cancer, prostate cancer, chronic lymphocytic leukemia, lymphoma, myeloma, and pancreatic cancer.

[0005] Myeloid cell leukemia 1 (Mcl-1), an anti-apoptotic Bcl-2 family member, is a regulator of cell survival. Amplification of the Mcl-1 gene and / or overexpression of the Mcl-1 protein has been observed in multiple cancer types and is associated with tumorigenesis (Beroukhim et al. (2010) Nature 463(7283):899-905). Mcl-1 is one of the most frequently amplified genes in human cancers and is also a critical survival factor that has been shown to mediate drug resistance to various anticancer drugs.

[0006] Mcl-1 is thought to promote cell survival by binding to pro-apoptotic proteins such as Bim, Noxa, Bak, and Bax and neutralizing their death-inducing activity. Inhibition of Mcl-1 releases these pro-apoptotic proteins and often induces apoptosis in tumor cells that depend on Mcl-1 for survival. Therefore, therapeutic targeting of Mcl-1 or its upstream and / or downstream proteins in the apoptotic signaling pathway may be a promising strategy for treating various malignancies and overcoming drug resistance in certain human cancers.

[0007] CD74 (DHLAG) is an established and attractive target for antibody-drug conjugates due to its restricted expression on normal tissues and significant upregulation in a range of hematological malignancies. CD74 functions as a chaperone required for assembly and transport of MHC class II complexes and as a receptor for macrophage migration inhibitory receptor (MIF). In oncology, it has been well documented that CD74 is significantly upregulated at both the RNA and protein levels in a range of B-cell and myeloid cell malignancies, including acute myeloid leukemia, multiple myeloma, and diffuse large B-cell lymphoma. In addition, CD74 is known to be rapidly internalized upon antibody association and transport to lysosomes, and to be rapidly relocated to the surface of tumor cells after internalization. Antibodies and antibody-drug conjugates targeting CD74 have previously been shown to exhibit antitumor activity in preclinical cancer models. Summary of the Invention

[0008] In some embodiments, the present disclosure provides, in part, novel antibody-drug conjugate (ADC) compounds that have biological activity against cancer cells. The compounds can slow, inhibit, and / or reverse tumor growth in mammals and / or may be useful in treating human cancer patients. More specifically, the present disclosure, in some embodiments, relates to ADC compounds that can bind and kill cancer cells. In some embodiments, the ADC compounds disclosed herein include a linker that connects an Mcl-1 inhibitor to a full-length anti-CD74 antibody or antigen-binding fragment. In some embodiments, the ADC compounds can also be internalized into target cells after binding.

[0009] In some embodiments, the ADC compound has formula (1): Ab-(LD) p (1) (wherein Ab is an anti-CD74 antibody or antigen-binding fragment thereof that targets cancer cells; D is an Mcl-1 inhibitor, L is a linker covalently linking Ab to D; p is an integer from 1 to 16. It can be expressed as:

[0010] In some embodiments, p is an integer from 1 to 8. In some embodiments, p is an integer from 1 to 5. In some embodiments, p is an integer from 2 to 4. In some embodiments, p is 2. In some embodiments, p is 4. In some embodiments, p is determined by liquid chromatography-mass spectrometry (LC-MS).

[0011] In some embodiments, the linker (L) comprises a linking group, at least one spacer group, and at least one cleavable group. In some cases, the cleavable group comprises a pyrophosphate group and / or a self-immolative group. In certain embodiments, L comprises a linking group, at least one bridging spacer group, and at least one cleavable group comprising a pyrophosphate group and / or a self-immolative group.

[0012] In some embodiments, the antibody-drug conjugate has the formula (A):

[0013] [ka] (In the formula, R 1 is a linking group, L is a bridging spacer group, and E is a cleavable group).

[0014] In some embodiments, the cleavable group comprises a pyrophosphate group.

[0015] [ka] Includes.

[0016] In some embodiments, the crosslinking spacer group comprises a polyoxyethylene (PEG) group. In some cases, the PEG group may be selected from PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, and PEG15. In some embodiments, the crosslinking spacer group may comprise -CO-CH2-CH2-PEG12-. In other embodiments, the crosslinking spacer group comprises a butanoyl, pentanoyl, hexanoyl, heptanoyl, or octanoyl group. In some embodiments, the crosslinking spacer group comprises a hexanoyl group.

[0017] In some embodiments, the linking group is formed from at least one reactive group selected from a maleimide group, a thiol group, a cyclooctyne group, and an azide group. For example, a maleimide group has the structure:

[0018] [ka] may have

[0019] The azide group has the structure: -N=N + =N - may have

[0020] The cyclooctyne group has the structure:

[0021] [ka] where

[0022] [ka] is the binding to the antibody.

[0023] In some cases, the cyclooctyne group has the structure:

[0024] [ka] where

[0025] [ka] is the binding to the antibody.

[0026] In some embodiments, the linking group is

[0027] [ka] and having a formula including:

[0028] [ka] is the binding to the antibody.

[0029] In some embodiments, the antibody

[0030] [ka] and is connected to the linker (L) by a linking group selected from where:

[0031] [ka] is the binding to the antibody, where

[0032] [ka] is the bond to the bridging spacer group.

[0033] In some embodiments, the bridging spacer group is linked to a cleavable group.

[0034] In some embodiments, the bridging spacer group is -CO-CH2-CH2-PEG12-.

[0035] In some embodiments, the cleavable group is -pyrophosphate-CH2-CH2-NH2-.

[0036] In some embodiments, the cleavable group is linked to the Mcl-1 inhibitor (D).

[0037] In some embodiments, the cleavable group is linked to the Mcl-1 inhibitor (D) group via a phenyl-pyrimidinyl group.

[0038] In some embodiments, the linker comprises a linking group, at least one bridging spacer group, a peptide group, and at least one cleavable group.

[0039] In some embodiments, the antibody-drug conjugate has formula (B):

[0040] [ka] (In the formula, R 1 is a linking group, L1 is a bridging spacer, Lp is a peptide group comprising 1 to 6 amino acid residues, E is a cleavable group, L2 is a bridging spacer, m is 0 or 1, and D is an Mcl-1 inhibitor. In some cases, m is 1 and the bridging spacer is

[0041] [ka] Includes.

[0042] In some embodiments, at least one crosslinking spacer comprises a PEG group. In some cases, the PEG group is selected from PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, and PEG15. In some cases, at least one crosslinking spacer is * -C(O)-CH2-CH2-PEG1- ** ,* -C(O)-CH2-PEG3- ** , * -C(O)-CH2-CH2-PEG12 ** , * -NH-CH2-CH2-PEG1- ** , polyhydroxyalkyl groups, and * -C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)- ** (In the formula, ** indicates the point of direct or indirect attachment of at least one bridge spacer to a linking group; * indicates the point of direct or indirect attachment of at least one cross-linking spacer to the peptide group).

[0043] In some embodiments, L1 is * -C(O)-CH2-CH2-PEG1- ** , * -C(O)-CH2-PEG3- ** , * -C(O)-CH2-CH2-PEG12 ** , * -NH-CH2-CH2-PEG1- ** and polyhydroxyalkyl groups, ** is R 1 indicates the direct or indirect point of attachment of L1 to * indicates the point of direct or indirect attachment of L1 to Lp).

[0044] In some embodiments, m is 1 and L2 is -C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)-.

[0045] In some embodiments, the peptide group comprises 1 to 12 amino acid residues. In some embodiments, the peptide group (Lp) comprises 1 to 10 amino acid residues. In some embodiments, the peptide group (Lp) comprises 1 to 8 amino acid residues. In some embodiments, the peptide group (Lp) comprises 1 to 6 amino acid residues. In some embodiments, the peptide group comprises 1 to 4 amino acid residues. In some embodiments, the peptide group comprises 1 to 3 amino acid residues. In some embodiments, the peptide group comprises 1 to 2 amino acid residues. In some cases, the amino acid residue is selected from L-glycine (Gly), L-valine (Val), L-citrulline (Cit), L-cysteic acid (sulfo-Ala), L-lysine (Lys), L-isoleucine (Ile), L-phenylalanine (Phe), L-methionine (Met), L-asparagine (Asn), L-proline (Pro), L-alanine (Ala), L-leucine (Leu), L-tryptophan (Trp), and L-tyrosine (Tyr). For example, the peptide group may include Val-Cit, Val-Ala, Val-Lys, and / or sulfo-Ala-Val-Ala. In some embodiments, the peptide group (Lp) is

[0046] [ka] In some embodiments, the peptide group (Lp) comprises one amino acid residue bound to a group:

[0047] [ka] The group includes a group selected from:

[0048] In some cases, the peptide group is

[0049] [ka] The group includes a group selected from:

[0050] In some embodiments, the self-immolative group comprises para-aminobenzyl-carbamate, para-aminobenzyl-ammonium, para-amino-(sulfo)benzyl-ammonium, para-amino-(sulfo)benzyl-carbamate, para-amino-(alkoxy-PEG-alkyl)benzyl-carbamate, para-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-carbamate, or para-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-ammonium.

[0051] In some embodiments, m is 1 and the bridging spacer is

[0052] [ka] Includes.

[0053] In some embodiments, the linker-drug moiety-(LD) is

[0054] [ka]

[0055] [ka]

[0056] [ka]

[0057] [ka] The compound is formed from a compound selected from:

[0058] In some embodiments, the antibody-drug conjugate comprises:

[0059] [ka]

[0060] [ka]

[0061] [ka]

[0062] [ka]

[0063] [ka] and where:

[0064] [ka] comprises a linker-drug group-(LD), which is the bond to the antibody.

[0065] In some embodiments, the antibody-drug conjugate has formula (C):

[0066] [ka] (In the formula, R 1 is a linking group, L1 is a bridging spacer, and L p is a peptide group containing 1 to 6 amino acids, D is an Mcl-1 inhibitor, G1-L2-A is a self-immolative spacer, L2 is a bond, methylene, neopentylene, or C2-C3 alkenylene, and A is a bond, -OC(=O)- * ,

[0067] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the bonding point with D, L3 is a spacer moiety, and R 2 is a hydrophilic moiety).

[0068] In some embodiments, the antibody-drug conjugate has formula (D):

[0069] [ka] (In the formula, R 1 is a linking group, L1 is a bridging spacer, Lp is a peptide group containing 1 to 6 amino acids, A is a bond, -OC(=O)- * ,

[0070] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the bonding point with D, L3 is a spacer moiety, and R 2 is a hydrophilic moiety).

[0071] In some embodiments, L1 is

[0072] [ka] or * -CH(OH)CH(OH)CH(OH)CH(OH)- ** wherein each n is an integer from 1 to 12, and wherein L1 * indicates the direct or indirect bond point with Lp, and ** is R 1 The point of direct or indirect attachment to the

[0073] In some embodiments, L1 is

[0074] [ka] and n is an integer from 1 to 12, where L1 * indicates the direct or indirect bond point with Lp, and ** is R 1 The point of direct or indirect attachment to the

[0075] In some embodiments, L1 is

[0076] [ka] and n is 1, where L1 * indicates the direct or indirect bond point with Lp, and ** is R 1 The point of direct or indirect attachment to the

[0077] In some embodiments, L1 is

[0078] [ka] and n is 12, where L1 * indicates the direct or indirect bond point with Lp, and ** is R 1 The point of direct or indirect attachment to the

[0079] In some embodiments, L1 is

[0080] [ka] and n is an integer from 1 to 12, where L1 * indicates the direct or indirect bond point with Lp, and ** is R 1 The point of direct or indirect attachment to the

[0081] In some embodiments, L1 is

[0082] [ka] where L1 * indicates the direct or indirect bond point with Lp, and ** is R 1 The point of direct or indirect attachment to the

[0083] In some embodiments, L1 is * -C(=O)(CH2) m O(CH2) m - ** ; * -C(=O)((CH2) m O) t (CH2) n - ** ; * -C(=O)(CH2) m - ** ; * -C(=O)NH((CH2) m O) t (CH2) n - ** ; * -C(=O)O(CH2) m SSC(R 3 )2(CH2) m C(=O)NR 3 (CH2) m NR 3C(=O)(CH2) m - ** ; * -C(=O)O(CH2) m C(=O)NH(CH2) m - ** ; * -C(=O)(CH2) m NH(CH2) m - ** ; * -C(=O)(CH2) m NH(CH2) n C(=O)- ** ; * -C(=O)(CH2) m X1(CH2) m - ** ; * -C(=O)((CH2) m O) t (CH2) n X1(CH2) n - ** ; * -C(=O)(CH2) m NHC(=O)(CH2) n - ** ; * -C(=O)((CH2) m O) t (CH2) n NHC(=O)(CH2) n - ** ; * -C(=O)(CH2) m NHC(=O)(CH2) n X1(CH2) n - ** ; * -C(=O)((CH2) m O) t (CH2) n NHC(=O)(CH2) n X1(CH2) n - ** ; * -C(=O)((CH2) m O) t (CH2)n C(=O)NH(CH2) m - ** ; * -C(=O)(CH2) m C(R 3 )2- ** or * -C(=O)(CH2) m C(=O)NH(CH2) m - ** wherein L1 is a bridging spacer comprising * indicates the direct or indirect bond point with Lp, and ** is R 1 where X1 represents a point of direct or indirect attachment to

[0084] [ka] and each m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each n is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; Each t is independently selected from 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 and 30.

[0085] In some embodiments, R 2 is polyethylene glycol, polyalkylene glycol, polyol, polysarcosine, sugar, oligosaccharide, polypeptide, or 1 to 3

[0086] [ka] In some embodiments, R is a hydrophilic moiety comprising a C2-C6 alkyl substituted with a group. 2 teeth,

[0087] [ka] (wherein n is an integer between 1 and 6),

[0088] [ka] is.

[0089] In some embodiments, the hydrophilic moiety has the formula:

[0090] [ka] (Wherein, R is H, —CH, —CHCHNHC(═O)OR a , -CH2CH2NHC(=O)R a or -CH2CH2C(=O)OR a and R' is OH, -OCH3, -CH2CH2NHC(=O)OR a , -CH2CH2NHC(=O)R a or -OCH2CH2C(=O)OR a where R a is H or OH or C 1~4 C optionally substituted with either alkoxy 1~4 alkyl, and each of m and n is an integer between 2 and 25 (e.g., between 3 and 25). Contains polyethylene glycol.

[0091] In some embodiments, the hydrophilic moiety is

[0092] [ka] Includes.

[0093] In some embodiments, the hydrophilic moiety is a polysarcosine, e.g., the moiety

[0094] [ka] (wherein n is an integer between 3 and 25, and R is H, -CH or -CHCHC(=O)OH) Polysarcosine containing:

[0095] In some embodiments, L3 has the structure

[0096] [ka] (In the formula, W is -CH2-, -CH2O-, -CH2N(R b )C(=O)O-, -NHC(=O)C(R b )2NHC(=O)O-, -NHC(=O)C(R b )2NH-, -NHC(=O)C(R b )2NHC(=O)-, -CH2N(XR 2 )C(=O)O-, -C(=O)N(XR 2 )-, -CH2N(XR 2 )C(=O)-, -C(=O)NR b -, -C(=O)NH-, -CH2NR b C(=O)-, -CH2NR b C(=O)NH-, -CH2NR b C(=O)NR b -, -NHC(=O)-, -NHC(=O)O-, -NHC(=O)NH-, -OC(=O)NH-, -S(O)2NH-, -NHS(O)2-, -C(=O)-, -C(=O)O- or -NH-, wherein each R b are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; X is a bond, triazolyl, or -CH2-triazolyl- is a spacer moiety having the following structure:

[0097] In some embodiments, L3 has the structure

[0098] [ka] (In the formula, W is -CH2-, -CH2O-, -CH2N(R b )C(=O)O-, -NHC(=O)C(R b )2NHC(=O)O-, -NHC(=O)C(R b )2NH-, -NHC(=O)C(R b )2NHC(=O)-, -CH2N(XR 2 )C(=O)O-, -C(=O)N(XR 2 )-, -CH2N(XR 2 )C(=O)-, -C(=O)NR b -, -C(=O)NH-, -CH2NR b C(=O)-, -CH2NR b C(=O)NH-, -CH2NR b C(=O)NR b -, -NHC(=O)-, -NHC(=O)O-, -NHC(=O)NH-, -OC(=O)NH-, -S(O)NH-, -NHS(O)-, -C(=O)-, -C(=O)O- or -NH-, wherein each R b are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl;

[0099] X is -CH2-triazolyl-C 1~4 Alkylene-OC(O)NHS(O)NH-, -C 4-6 Cycloalkylene -OC(O)NHS(O)2NH-, -(CH2CH2O) n -C(O)NHS(O)2NH-, -(CH2CH2O) n -C(O)NHS(O)2NH-(CH2CH2O) n -or -CH2-triazolyl-C 1~4 Alkylene-OC(O)NHS(O)2NH-(CH2CH2O) n where each n is independently 1, 2, or 3. is a spacer moiety having the following structure:

[0100] In some embodiments, the linking group is formed by a reaction involving at least one reactive group. In some cases, the linking group is formed by reacting a first reactive group that is attached to a linker with a second reactive group that is attached to an antibody or is an amino acid residue of an antibody.

[0101] In some embodiments, at least one of the reactive groups is thiols, Maleimide, haloacetamides, Azide, Alkynes, cyclooctene, triarylphosphines, oxanorbornadiene, cyclooctyne, diaryltetrazines, monoaryltetrazines, norbornene, aldehyde, hydroxylamine, hydrazine, NH2-NH-C(=O)-, ketones, vinyl sulfone, Aziridine, amino acid residues,

[0102] [ka] , -ONH2, -NH2,

[0103] [ka] , -N3,

[0104] [ka] , -SH, -SR 3 , -SSR 4, -S(=O)2(CH=CH2), -(CH2)2S(=O)2(CH=CH2), -NHS(=O)2(CH=CH2), -NHC(=O)CH2Br, -NHC(=O)CH2I,

[0105] [ka] , -C(O)NHNH2,

[0106] [ka]

[0107] [ka] Including, where: Each R 3 are independently selected from H and C1-C6 alkyl; Each R 4 is 2-pyridyl or 4-pyridyl, Each R 5 are independently selected from H, C1-C6 alkyl, F, Cl, and —OH; Each R 6 are independently selected from H, C1-C6 alkyl, F, Cl, —NH2, —OCH3, —OCH2CH3, —N(CH3)2, —CN, —NO2, and —OH; Each R 7 is H, C 1~6 Alkyl, fluoro, benzyloxy substituted with -C(=O)OH, benzyl substituted with -C(=O)OH, C substituted with -C(=O)OH 1~4 C substituted with alkoxy and -C(=O)OH 1~4 alkyl.

[0108] In some embodiments, the first reactive group and the second reactive group are thiols and maleimides, thiols and haloacetamides, thiols and vinyl sulfones, thiols and aziridines, Azides and alkynes, azide and cyclooctyne, azide and cyclooctene, azides and triarylphosphines, azides and oxanorbornadienes, diaryltetrazines and cyclooctene, monoaryltetrazines and norbornenes, aldehydes and hydroxylamines, aldehydes and hydrazines, Aldehydes and NH2-NH-C(=O)-, ketones and hydroxylamines, ketones and hydrazines, ketones and NH2-NH-C(=O)-, Hydroxylamine and

[0109] [ka] , amines and

[0110] [ka] ,or CoA or CoA analogue and serine residue Includes.

[0111] In some embodiments, the linking group is

[0112] [ka]

[0113] [ka]

[0114] [ka]

[0115] [ka] and where: R 32 is H, C 1~4 alkyl, phenyl, pyrimidine or pyridine; R 35 is H, C 1~6 C substituted with alkyl, phenyl, or 1 to 3 -OH groups 1~4 is alkyl, Each R 7 is H, C 1~6 Alkyl, fluoro, benzyloxy substituted with -C(=O)OH, benzyl substituted with -C(=O)OH, C substituted with -C(=O)OH 1~4 C substituted with alkoxy and -C(=O)OH 1~4 independently selected from alkyl, R 37 are independently selected from H, phenyl and pyridine; q is 0, 1, 2 or 3; R 8 is H or methyl, R 9 is H, —CH3 or phenyl.

[0116] In some embodiments, the peptide group (Lp) comprises 1 to 6 amino acid residues. In some embodiments, the peptide group (Lp) comprises 1 to 4 amino acid residues. In some embodiments, the peptide group comprises 1 to 3 amino acid residues. In some embodiments, the peptide group comprises 1 to 2 amino acid residues. In some embodiments, the amino acid residues are selected from L-glycine (Gly), L-valine (Val), L-citrulline (Cit), L-cysteic acid (sulfo-Ala), L-lysine (Lys), L-isoleucine (Ile), L-phenylalanine (Phe), L-methionine (Met), L-asparagine (Asn), L-proline (Pro), L-alanine (Ala), L-leucine (Leu), L-tryptophan (Trp), and L-tyrosine (Tyr). In some embodiments, the peptide group comprises Val-Cit, Phe-Lys, Val-Ala, Val-Lys, Leu-Cit, sulfo-Ala-Val and / or sulfo-Ala-Val-Ala.

[0117] [ka] is selected from.

[0118] In some embodiments, the linker-drug group-(LD) has the formula:

[0119] [ka] (In the formula, R is H, -CH or -CHCHC(=O)OH; A is a bond, -OC(=O)- * ,

[0120] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a)2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0121] In some embodiments, the linker-drug group-(LD) has the formula:

[0122] [ka] (In the formula, R is H, -CH or -CHCHC(=O)OH; A is a bond, -OC(=O)- * ,

[0123] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0124] In some embodiments, the linker-drug group-(LD) has the formula:

[0125] [ka] (In the formula, R is H, -CH or -CHCHC(=O)OH; A is a bond, -OC(=O)- * ,

[0126] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0127] In some embodiments, the linker-drug group-(LD) has the formula:

[0128] [ka] (In the formula, each R is independently selected from H, —CH, and —CHCHC(═O)OH; A is a bond, -OC(=O)- * ,

[0129] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; *indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0130] In some embodiments, the linker-drug group-(LD) has the formula:

[0131] [ka] (In the formula, each R is independently selected from H, —CH, and —CHCHC(═O)OH; A is a bond, -OC(=O)- * ,

[0132] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0133] In some embodiments, the linker-drug group-(LD) has the formula:

[0134] [ka] (In the formula, Xa is -CH2-, -OCH2-, -NHCH2-, or -NRCH2-, and each R is independently H, -CH3, or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- * ,

[0135] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0136] In some embodiments, the linker-drug group-(LD) has the formula:

[0137] [ka] (In the formula, R is H, -CH or -CHCHC(=O)OH; A is a bond, -OC(=O)- * ,

[0138] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0139] In some embodiments, the linker-drug group-(LD) has the formula:

[0140] [ka] (In the formula, Xb is -CH2-, -OCH2-, -NHCH2-, or -NRCH2-, and each R is independently H, -CH3, or -CH2CH2C(=O)OH; A is a bond, -OC(=O)- * ,

[0141] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0142] In some embodiments, the linker-drug group-(LD) has the formula:

[0143] [ka] (In the formula, A is a bond, -OC(=O)- * ,

[0144] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- *or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0145] In some embodiments, the linker-drug group-(LD) has the formula:

[0146] [ka] (In the formula, A is a bond, -OC(=O)- * ,

[0147] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0148] In some embodiments, the linker-drug group-(LD) has the formula:

[0149] [ka] (In the formula, A is a bond, -OC(=O)-* ,

[0150] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0151] In some embodiments, the linker-drug group-(LD) has the formula:

[0152] [ka] (In the formula, A is a bond, -OC(=O)- * ,

[0153] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0154] In some embodiments, the linker-drug group-(LD) has the formula:

[0155] [ka] (In the formula, A is a bond, -OC(=O)- * ,

[0156] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0157] In some embodiments, the linker-drug group-(LD) has the formula:

[0158] [ka] (In the formula, A is a bond, -OC(=O)- * ,

[0159] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R aare independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0160] In some embodiments, the linker-drug group-(LD) has the formula:

[0161] [ka] (In the formula, A is a bond, -OC(=O)- * ,

[0162] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0163] In some embodiments, the linker-drug group-(LD) has the formula:

[0164] [ka] (In the formula, each R is independently H, —CH, or —CHCHC(═O)OH; A is a bond, -OC(=O)- * ,

[0165] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * and Here, each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, D is an Mcl-1 inhibitor) The compound comprises or is formed from:

[0166] In some embodiments, A is a bond.

[0167] In some embodiments, R is —CH 3 .

[0168] In some embodiments, the Mcl-1 inhibitor (D) has the formula (I):

[0169] [ka] (In the formula, Ring D0 is a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group; Ring E0 is a furyl, thienyl or pyrrolyl ring; X 01 , X 03 , X 04 and X 05 are each independently a carbon atom or a nitrogen atom, X 02 is CR 026 group or a nitrogen atom,

[0170] [ka] means that the ring is aromatic, Y0 is a nitrogen atom or CR 03 It is the basis, Z0 is a nitrogen atom or CR 04 It is the basis, R 01 is a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl group, a hydroxy group, a hydroxy(C1-C6) alkyl group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, -Cy 08 , -(C0-C6) alkyl-NR 011 R 011 ', -O-(C1-C6) alkyl-NR 011 R 011 ', -O-(C1-C6) alkyl-R 012 , -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ' or -SO2-(C1-C6)alkyl; R 02 , R 03 , R 04 and R 05 are each independently a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl, a hydroxy group, a hydroxy(C1-C6) alkyl group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, a -(C0-C6) alkyl-NR 011 R 011 ', -O-Cy01 , -(C0-C6) alkyl-Cy 01 , -(C2-C6)alkenyl-Cy 01 , -(C2-C6)alkynyl-Cy 01 , -O-(C1-C6) alkyl-NR 011 R 011 ', -O-(C1-C6) alkyl-R 031 , -O-(C1-C6)alkyl-R 012 , -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ',or -SO2-(C1-C6) alkyl, Or pair (R 01 , R 02 ), (R 02 , R 03 ), (R 03 , R 04 ) or (R 04 , R 05 ) together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring may be substituted with halogen, linear or branched (C1-C6) alkyl, (C0-C6) alkyl-NR 011 R 011 ', -NR 013 R 013 ', -(C0-C6) alkyl-Cy 01 or oxo; R 06 and R 07are each independently a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl, a hydroxy group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, or a -(C0-C6) alkyl-NR 011 R 011 ', -O-(C1-C6) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C0-C6) alkyl-Cy 01 , -(C2-C6)alkenyl-Cy 01 , -(C2-C6)alkynyl-Cy 01 , -O-(C1-C6) alkyl-R 012 , -C(O)-OR 11 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 11 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ', or -SO2-(C1-C6) alkyl; Or pair (R 06 , R 07 ), when fused with two adjacent carbon atoms, together with the carbon atoms to which they are attached, form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring can be optionally fused with a linear or branched (C1-C6) alkyl group, -NR 013 R 013 ', -(C0-C6) alkyl-Cy 01 or optionally substituted with oxo, W0 is a -CH2- group, a -NH- group or an oxygen atom; R 08 is a hydrogen atom, a linear or branched (C1-C8) alkyl group, -CHR 0a R 0b a group, an aryl group, a heteroaryl group, an aryl(C1-C6)alkyl group, or a heteroaryl(C1-C6)alkyl group, R 09 is a hydrogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, -Cy 02 , -(C1-C6) alkyl-Cy 02 , -(C2-C6)alkenyl-Cy 02 , -(C2-C6)alkynyl-Cy 02 , -Cy 02 -Cy 03 , -(C2-C6)alkynyl-O-Cy 02 , -Cy 02 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 03 , halogen atoms, cyano groups, -C(O)-R 014 or -C(O)-NR 014 R 014 ' and R 010 is a hydrogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, an aryl (C1-C6) alkyl group, a (C1-C6) cycloalkyl alkyl group, a linear or branched (C1-C6) haloalkyl group, or -(C1-C6) alkyl-O-Cy 04 and Or pair (R 09 , R 010 ), when fused with two adjacent carbon atoms, together with the carbon atoms to which they are attached, form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains 1 to 3 heteroatoms selected from O, S and N; R 011 and R 011' are each independently a hydrogen atom, an optionally substituted linear or branched (C1-C6) alkyl group, or -(C0-C6) alkyl-Cy 01 and Or pair (R 011 , R 011 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the nitrogen may be substituted with one or two groups selected from a hydrogen atom and a linear or branched (C1-C6) alkyl group, wherein one or more of the carbon atoms of the linear or branched (C1-C6) alkyl group are optionally deuterated; R 012 -Cy 05 , -Cy 05 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 06 , -Cy 05 -(C0-C6) alkyl-Cy 06 , -Cy 05 -(C0-C6) alkyl-NR 011 -(C0-C6) alkyl-Cy 06 , -Cy 05 -Cy 06 -O-(C0-C6) alkyl-Cy 07 , -Cy 05 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 09 , -Cy 05 -(C0-C6) alkyl-Cy 09 , -NH-C(O)-NH-R 011 , -Cy 05 -(C0-C6) alkyl-NR 011 -(C0-C6) alkyl-Cy 09 , -C(O)-NR 011 R 011 ', -NR 011 R 011 ', -OR 011 , -NR 011 -C(O)-R011 ', -O-(C1-C6) alkyl-OR 011 , -SO2-R 011 , and -C(O)-OR 011 and R 013 , R 013 ', R 014 and R 014 ' are each independently a hydrogen atom or an optionally substituted linear or branched (C1-C6) alkyl group, R 0a is a hydrogen atom or a linear or branched (C1-C6) alkyl group, R 0b is -OC(O)-OR 0c group, -OC(O)-NR 0c R 0c ' group or -OP(O)(OR 0c )2 units, R 0c and R 0c ' are each independently a hydrogen atom, a linear or branched (C1-C8) alkyl group, a cycloalkyl group, a (C1-C6) alkoxy(C1-C6) alkyl group or a (C1-C6) alkoxycarbonyl(C1-C6) alkyl group, Or pair (R 0c , R 0c ') together with the nitrogen atom to which they are attached form a non-aromatic ring consisting of 5 to 7 ring members which, in addition to the nitrogen atom, may contain 1 to 3 heteroatoms selected from oxygen and nitrogen, wherein the nitrogen is optionally substituted with a linear or branched (C1-C6) alkyl group; Cy 01 , Cy 02 , Cy 03 , Cy 04 , Cy 05 , Cy 06 , Cy 07 , Cy 08 and Cy 010 represent, independently of each other, a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group, each of which is optionally substituted; Cy 09 teeth

[0171] [ka] and or Cy 09 is -OP(O)(OR 020 )2;-OP(O)(O - M + )2;-(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 ;Hydroxy;Hydroxy(C1-C6)alkyl;-(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl; and -U0-(CH2) q0 -NR 021 R 021 ', and R 015 is a hydrogen atom; -(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 Group; Linear or branched (C1-C6) alkoxy (C1-C6) alkyl group; -U0-(CH2) q0 -NR 021 R 021 ' group; or -(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl group, R 016 represents a hydrogen atom; a hydroxy group; a hydroxy(C1-C6) alkyl group; -(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl group; (CH2) r0 -U0-V0-OP(O)(OR 020 ) 2 groups;-OP(O)(O - M + )2 groups;-OS(O)2OR 020 Group;-S(O)2OR 020 Group;-(CH2) p0 -O-(CHR018 -CHR 019 -O) q0 -R 020 Group;-(CH2) p0 -OC(O)-NR 022 R 023 group; or -U0-(CH2) q0 -NR 021 R 021 'Based on R 017 is a hydrogen atom; -(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 Group: -CH2-P(O)(OR 020 ) 2 units, -OP(O)(OR 020 ) 2 groups;-OP(O)(O - M + )2 group; hydroxy group; hydroxy(C1-C6) alkyl group; -(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl group; -U0-(CH2) q0 -NR 021 R 021 ' group; or aldonic acid, M + is a pharmaceutically acceptable monovalent cation, U0 is a bond or an oxygen atom, V0 is -(CH2) s0 - group or -C(O)- group, R 018 is a hydrogen atom or a (C1-C6)alkoxy(C1-C6)alkyl group, R 019 is a hydrogen atom or a hydroxy(C1-C6)alkyl group, R 020 is a hydrogen atom or a linear or branched (C1-C6) alkyl group, R 021 and R 021 ' are each independently a hydrogen atom, a linear or branched (C1-C6) alkyl group, or a hydroxy(C1-C6) alkyl group, Or pair (R 021 , R021 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring is optionally substituted with hydrogen atoms or linear or branched (C1-C6) alkyl groups; R 022 represents a (C1-C6) alkoxy(C1-C6) alkyl group, -(CH2) p0 -NR 024 R 024 ' group or -(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 It is the basis, R 023 is a hydrogen atom or a (C1-C6)alkoxy(C1-C6)alkyl group, Or pair (R 022 , R 023 ) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 18 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 5 heteroatoms selected from O, S and N, wherein the resulting ring is optionally substituted with a hydrogen atom, a linear or branched (C1-C6) alkyl group or a heterocycloalkyl group; R 024 and R 024 ' are each independently a hydrogen atom or a linear or branched (C1-C6) alkyl group, Or pair (R 024 , R 024 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring consisting of 5 to 7 ring members, which may contain, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring is optionally substituted with hydrogen atoms or linear or branched (C1-C6) alkyl groups; R 025 is a hydrogen atom, a hydroxy group, or a hydroxy(C1-C6)alkyl group, R026 is a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, or a cyano group, R 027 is a hydrogen atom or a linear or branched (C1-C6) alkyl group, R 028 is -OP(O)(O - )(O - ) group, -OP(O)(O - )(OR 030 ) group, -OP(O)(OR 030 )(OR 030 ') group, -(CH2) p0 -O-SO2-O - Group, -(CH2) p0 -SO2-O - Group, -(CH2) p0 -O-SO2-OR 030 Group, -Cy 010 , -(CH2) p0 -SO2-OR 030 group, -OC(O)-R 029 The group -OC(O)-OR 029 group or -OC(O)-NR 029 R 029 'Based on R 029 and R 029 ' are each independently a hydrogen atom, a linear or branched (C1-C6) alkyl group or a linear or branched amino(C1-C6) alkyl group, R 030 and R 030 ' are each independently a hydrogen atom, a linear or branched (C1-C6) alkyl group or an aryl (C1-C6) alkyl group, R 031 teeth

[0172] [ka] wherein the ammonium cation optionally exists in zwitterionic form or has a monovalent anionic counterion; n0 is an integer equal to 0 or 1, p0 is an integer equal to 0, 1, 2, or 3; q0 is an integer equal to 1, 2, 3, or 4; r0 and s0 are independently integers equal to 0 or 1; Here, at most, R 03 Group, R 09 group or R 012 One of the groups, if present, is covalently attached to the linker, where the valency of the atom is not exceeded by the one or more substituents attached to it. or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0173] In some embodiments, Cy 01 , Cy 02 , Cy 03 , Cy 04 , Cy 05 , Cy 06 , Cy 07 , Cy 08 and Cy 010 are, independently of each other, a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group, each of which is selected from the group consisting of halo; -(C1-C6)alkoxy; -(C1-C6)haloalkyl; -(C1-C6)haloalkoxy; -(CH2) p0 -O-SO2-OR 030 ; -(CH2) p0 -SO2-OR 030 ;-OP(O)(OR 020 )2;-OP(O)(O - M + )2;-CH2-P(O)(OR 020 )2; -(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 ;Hydroxy;Hydroxy(C1-C6)alkyl; -(CH2) r0 -U0-(CH2) s0-heterocycloalkyl; or -U0-(CH2) q0 -NR 021 R 021 Optionally substituted with one or more groups selected from:

[0174] In some embodiments, D is a group represented by formula (II):

[0175] [ka] (In the formula, Z0 is a nitrogen atom or CR 04 It is the basis, R 01 is a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl group, a hydroxy group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, -Cy 08 , -NR 011 R 011 ' and R 02 , R 03 and R 04 are each independently a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl, a hydroxy group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, or a -(C0-C6) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C0-C6) alkyl-Cy 01 , -(C2-C6)alkenyl-Cy 01 , -(C2-C6)alkynyl-Cy 01 , -O-(C1-C6) alkyl-NR 011 R 011 ', -O-(C1-C6)alkyl-R031 , -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ',or -SO2-(C1-C6) alkyl, Or pair (R 02 , R 03 ) or (R 03 , R 04 ) together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, wherein the ring is selected from linear or branched (C1-C6) alkyl, -NR 013 R 013 ', -(C0-C6) alkyl-Cy 01 and oxo, R 06 and R 07 are each independently a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl, a hydroxy group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, or a -(C0-C6) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C0-C6) alkyl-Cy 01 , -(C2-C6)alkenyl-Cy 01 , -(C2-C6)alkynyl-Cy 01 , -O-(C1-C6) alkyl-R 012, -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ', or -SO2-(C1-C6) alkyl; Or pair (R 06 , R 07 ), when fused with two adjacent carbon atoms, together with the carbon atoms to which they are attached, form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring can be optionally fused with a linear or branched (C1-C6) alkyl group, -NR 013 R 013 ', -(C0-C6) alkyl-Cy 01 and oxo, R 08 is a hydrogen atom, a linear or branched (C1-C8) alkyl group, an aryl group, a heteroaryl group, an aryl-(C1-C6) alkyl group or a heteroaryl-(C1-C6) alkyl group, R 09 is a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, -Cy 02 , -(C1-C6) alkyl-Cy 02 , -(C2-C6)alkenyl-Cy 02 , -(C2-C6)alkynyl-Cy 02 , -Cy 02 -Cy 03 , -(C2-C6)alkynyl-O-Cy 02 , -Cy 02 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 03, halogen atoms, cyano groups, -C(O)-R 014 , -C(O)-NR 014 R 014 ' and R 011 and R 011 ' are each independently a hydrogen atom, an optionally substituted linear or branched (C1-C6) alkyl group, or -(C0-C6) alkyl-Cy 01 and Or pair (R 011 , R 011 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the N atom is optionally substituted with a linear or branched (C1-C6) alkyl group, and wherein one or more of the carbon atoms of the linear or branched (C1-C6) alkyl group is optionally deuterated; R 012 -Cy 05 , -Cy 05 -(C0-C6) alkyl-Cy 06 , -Cy 05 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 06 , -Cy 05 -(C0-C6) alkyl-NR 011 -(C0-C6) alkyl-Cy 06 , -Cy 05 -Cy 06 -O-(C0-C6) alkyl-Cy 07 , -Cy 05 -(C0-C6) alkyl-Cy 09 , -NH-C(O)-NH-R 011 , -C(O)-NR 011 R 011 ', -NR 011 R 011 ', -OR 011 , -NR 011 -C(O)-R 011 ', -O-(C1-C6) alkyl-OR 011 , -SO2-R 011 , or -C(O)-OR 011 represents R 013 , R 013 ', R 014 and R 014 ' are each independently a hydrogen atom or an optionally substituted linear or branched (C1-C6) alkyl group; Cy 01 , Cy 02 , Cy 03 , Cy 05 , Cy 06 , Cy 07 and Cy 08 are, independently of each other, an optionally substituted cycloalkyl group, an optionally substituted heterocycloalkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group; Cy 09 teeth

[0176] [ka] and where R 015 , R 016 and R 017 is as defined for formula (I), R 031 teeth

[0177] [ka] where R 027 and R 028 is as defined for formula (I), Here, at most, R 03 Group, R 09 group or R 012 One of the groups, if present, is covalently attached to the linker or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0178] In some embodiments, D is a group of formula (III):

[0179] [ka] (In the formula, R 01 is a linear or branched (C1-C6) alkyl group, R 03 is -O-(C1-C6) alkyl-NR 011 R 011 'or

[0180] [ka] and where R 011 and R 011 ' are each independently a hydrogen atom, an optionally substituted linear or branched (C1-C6) alkyl group, or -(C0-C6) alkyl-Cy 01 and Or pair (R 011 , R 011 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the N atom may be substituted with one or two groups selected from a hydrogen atom or a linear or branched (C1-C6) alkyl group; where R 027 is a hydrogen atom, and R 028 is -(CH2) p0 -O-SO2-O - group or -(CH2) p0 -SO2-OR 030 It is the basis, R 09 is a linear or branched (C2-C6) alkynyl group or -Cy 02 and R 012 -Cy 05 , -Cy 05-(C0-C6) alkyl-Cy 06 or -Cy 05 -(C0-C6) alkyl-Cy 09 and Cy 01 , Cy 02 , Cy 05 and Cy 06 are, independently of each other, a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group, each of which is optionally substituted; Cy 09 teeth

[0181] [ka] and R 015 , R 016 and R 017 is as defined for formula (I), Here, at most, R 03 Group, R 09 group or R 012 One of the groups, if present, is covalently attached to the linker or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0182] In some embodiments, Cy 01 , Cy 02 , Cy 05 , Cy 06 are, independently of each other, a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group, each of which is selected from the group consisting of halo; -(C1-C6)alkoxy; -(C1-C6)haloalkyl; -(C1-C6)haloalkoxy; -(CH2) p0 -O-SO2-OR 030 ;-(CH2) p0 -SO2-OR 030 ; -OP(O)(OR 020 )2;-OP(O)(O - M +)2;-CH2-P(O)(OR 020 )2; -(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 ;Hydroxy;Hydroxy(C1-C6)alkyl; -(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl; or -U0-(CH2) q0 -NR 021 R 021 Optionally substituted with one or more groups selected from:

[0183] In some embodiments, R 01 is methyl or ethyl.

[0184] In some embodiments, R 03 is -O-CH2-CH2-NR 011 R 011’ (In the formula, R 011 and R 011 ' together with the nitrogen atom that carries them form a piperazinyl group which may be substituted with a hydrogen atom or a linear or branched (C1-C6) alkyl group).

[0185] In some embodiments, R 03 is the expression:

[0186] [ka] (In the formula, R 027 is a hydrogen atom, and R 028 Ha-(CH2) p0 -O-SO2-OR 030 group, and p0 is an integer equal to 0, 1, 2, or 3, where R 030 represents a hydrogen atom, a linear or branched (C1-C6) alkyl group, or an aryl (C1-C6) alkyl group. Includes.

[0187] In some embodiments, R 03 is the expression:

[0188] [ka] (In the formula,

[0189] [ka] is the bond to the linker) Includes.

[0190] In some embodiments, Cy 01 , Cy 02 , Cy 03 , Cy 04 , Cy 05 , Cy 06 , Cy 07 , Cy 08 and Cy 010 are each independently an optionally substituted cycloalkyl group, an optionally substituted heterocycloalkyl group, an optionally substituted aryl group or an optionally substituted heteroaryl group, wherein the optional substituents are optionally substituted linear or branched (C1-C6) alkyl, optionally substituted linear or branched (C2-C6) alkenyl group, optionally substituted linear or branched (C2-C6) alkynyl group, optionally substituted linear or branched (C1-C6) alkoxy, optionally substituted (C1-C6) alkyl-S-, hydroxyl ... and R is selected from oxy, oxo (or N-oxide, where appropriate), nitro, cyano, —C(O)—OR′, —OC(O)—R′, —C(O)—NR′R′, —NR′R′, —(C═NR′)—OR′, linear or branched (C1-C6) haloalkyl, trifluoromethoxy, or halogen, wherein R′ and R′ are each independently a hydrogen atom or an optionally substituted linear or branched (C1-C6) alkyl group, and wherein one or more of the carbon atoms of the linear or branched (C1-C6) alkyl group is optionally deuterated.

[0191] In some embodiments, R 09 Cy 02 group, preferably an aryl group, more preferably a phenyl group. 02 is an optionally substituted aryl group.

[0192] In some embodiments, Cy 05 comprises a heteroaryl group selected from pyrazolyl and pyrimidinyl groups.

[0193] In some embodiments, Cy 05 is a pyrimidinyl group.

[0194] In some embodiments, Cy 05 is a pyrimidinyl group, and Cy 06 is a phenyl group.

[0195] In some embodiments, the linker (L) is a linker from L to R of formula (I), (II), or (III). 03 In some embodiments, the linker (L) is connected to D by a covalent bond from L to R of formula (I), (II), or (III). 09 is covalently bonded to D.

[0196] In some embodiments, D is

[0197] [ka]

[0198] [ka] or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0199] In some embodiments, -(LD) is formed from a compound selected from Table A or an enantiomer, diastereoisomer, atropisomer, deuterated derivative, and / or a pharmaceutically acceptable salt thereof. For the compounds in Table A, depending on their electronic charge, these compounds may be substituted with one pharmaceutically acceptable monovalent anionic counterion, M1. - In some embodiments, a monovalent anionic counterion M1 - may be selected from bromide, chloride, iodide, acetate, trifluoroacetate, benzoate, mesylate, tosylate, triflate, formate, or the like. In some embodiments, the monovalent anionic counterion M - is trifluoroacetic acid or formic acid.

[0200] [Table 1-1]

[0201] [Table 1-2]

[0202] [Table 1-3]

[0203] [Table 1-4]

[0204] [Table 1-5]

[0205] [Table 1-6]

[0206] [Table 1-7]

[0207] Table 1-8

[0208] Table 1-9

[0209] Table 1-10

[0210] Table 1-11

[0211] Table 1-12

[0212] Table 1-13

[0213] Table 1-14

[0214] Table 1-15

[0215] Table 1-16

[0216] Table 1-17

[0217] Table 1-18

[0218] Table 1-19

[0219] Table 1-20

[0220] Table 1-21

[0221] Table 1-22

[0222] Table 1-23

[0223] Table 1-24

[0224] Table 1-25

[0225] Table 1-26

[0226] Table 1-27

[0227] Table 1-28

[0228] Table 1-29

[0229] Table 1-30

[0230] Table 1-31

[0231] Table 1-32

[0232] Table 1-33

[0233] Table 1-34

[0234] Table 1-35

[0235] Table 1-36

[0236] Table 1-37

[0237] [Table 1-38]

[0238] [Table 1-39]

[0239] [Table 1-40]

[0240] [Table 1-41]

[0241] [Table 1-42]

[0242] [Table 1-43]

[0243] [Table 1-44]

[0244] [Table 1-45]

[0245] In some embodiments, the antibody-drug conjugate has a formula according to any one of the structures shown in Table B.

[0246] [Table 2-1]

[0247] [Table 2-2]

[0248] Table 2-3

[0249] Table 2-4

[0250] Table 2-5

[0251] Table 2-6

[0252] Table 2-7

[0253] Table 2-8

[0254] Table 2-9

[0255] Table 2-10

[0256] Table 2-11

[0257] Table 2-12

[0258] As used herein, "L / P" refers to a linker-payload, linker-drug, or linker-compound disclosed herein, and the terms "L#-P#" and "L#-C#" are used interchangeably to refer to a particular linker-drug disclosed herein, while the codes "P#" and "C#" are used interchangeably to refer to a particular compound unless otherwise specified. For example, "L1-C1" and "L1-P1" both refer to the same linker-payload structure disclosed herein, while "C1" and "P1" both refer to the same compound disclosed herein, including enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0259] In some embodiments, the antibody or antigen-binding fragment binds to the target antigen CD74 on cancer cells. In some embodiments, the CD74 is a human CD74 isoform. In some embodiments, the human CD74 isoform is

[0260] [ka] The isoform 1 (NP_001020330.1) has the amino acid sequence:

[0261] In some embodiments, the human CD74 isoform is

[0262] [ka] The isoform 2 (NP_004346.1) has the amino acid sequence:

[0263] Also provided herein, in some embodiments, are compositions comprising multiple copies of an antibody-drug conjugate (e.g., any of the exemplary antibody-drug conjugates described herein). In some embodiments, the average p of the antibody-drug conjugates in the composition is about 2 to about 4.

[0264] Also provided herein, in some embodiments, is a pharmaceutical composition comprising an antibody-drug conjugate (e.g., any of the exemplary antibody-drug conjugates described herein) or composition (e.g., any of the exemplary compositions described herein) and a pharmaceutically acceptable carrier.

[0265] Further provided herein, in some embodiments, are therapeutic uses for the described ADC compounds and compositions, e.g., in the treatment of cancer. In some embodiments, the present disclosure provides methods of treating cancer (e.g., a cancer that expresses the CD74 antigen targeted by an antibody or antigen-binding fragment of an ADC). In some embodiments, the present disclosure provides methods of reducing or slowing the expansion of a cancer cell population in a subject. In some embodiments, the present disclosure provides methods of determining whether a subject having or suspected of having cancer will respond to treatment with an ADC compound or composition disclosed herein.

[0266] An exemplary embodiment is a method of treating a subject having or suspected of having cancer, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein). In some embodiments, the cancer expresses the target antigen CD74. In some embodiments, the cancer is a tumor or blood cancer. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[0267] Another exemplary embodiment is a method of reducing or inhibiting tumor growth in a subject, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein). In some embodiments, the tumor expresses the target antigen CD74. In some embodiments, the tumor is breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular carcinoma, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the tumor is gastric cancer. In some embodiments, administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces or inhibits tumor growth by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.

[0268] Another exemplary embodiment is a method of reducing or slowing the expansion of a cancer cell population in a subject, comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein). In some embodiments, the cancer cell population expresses the target antigen CD74. In some embodiments, the cancer cell population is derived from a tumor or hematological cancer. In some embodiments, the cancer cell population is derived from breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer cell population is derived from lymphoma or gastric cancer. In some embodiments, administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%. In some embodiments, administration of the antibody-drug conjugate, composition, or pharmaceutical composition slows the expansion of the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%.

[0269] Another exemplary embodiment is an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) for use in treating a subject having or suspected of having cancer. In some embodiments, the cancer expresses the target antigen CD74. In some embodiments, the cancer is a tumor or blood cancer. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[0270] Another exemplary embodiment is the use of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) in the treatment of a subject having or suspected of having cancer. In some embodiments, the cancer expresses the target antigen CD74. In some embodiments, the cancer is a tumor or a blood cancer. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[0271] Another exemplary embodiment is the use of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) in a method of manufacturing a medicament for treating a subject having or suspected of having cancer. In some embodiments, the cancer expresses the target antigen CD74. In some embodiments, the cancer is a tumor or blood cancer. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[0272] Another exemplary embodiment is a method of determining whether a subject having or suspected of having cancer will respond to treatment with an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any of the exemplary antibody-drug conjugates, compositions, or pharmaceutical compositions disclosed herein) by providing a biological sample from the subject; contacting the sample with an antibody-drug conjugate; and detecting binding of the antibody-drug conjugate to cancer cells in the sample. In some embodiments, the cancer cells in the sample express the target antigen CD74. In some embodiments, the cancer expresses the target antigen CD74. In some embodiments, the cancer is a tumor or a blood cancer. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer is lymphoma or gastric cancer. In some embodiments, the sample is a tissue biopsy sample, a blood sample, or a bone marrow sample.

[0273] Also disclosed are methods of making the described ADC compounds and compositions. An exemplary embodiment is a method of making an antibody-drug conjugate by reacting an antibody or antigen-binding fragment with a cleavable linker linked to an Mcl-1 inhibitor under conditions that allow conjugation. [Brief explanation of the drawings]

[0274] [Figure 1A] FIG. 1 shows in vitro activity (CTG 72 h) of CD74-L7-P1 and P1 payload in DLBCL cell lines. [Figure 1B] FIG. 1 shows in vitro activity (CTG 72 h) of CD74-L7-P1, IgG-L7-P1 and P1 payloads in DLBCL cell lines. [Figure 2] FIG. 1 shows the in vitro activity (CTG 72 h) of various CD74 ADCs and corresponding payloads in the Monomac1 cell line. [Figure 3] Figure 1 shows the in vitro activity of CD74 MCL-1 antibody drug conjugate CD74-L7-P1, isotype IgG-L7-P1 ADC, and MCL-1 free payload P1 against endogenous cancer cell lines including KMS-21BM, KMS-20, MONO-MAC-1, NOMO-1, Kasumi-6, and EOL-1. [Figure 4] FIG. 1 shows the in vitro activity of CD74 MCL-1 antibody drug conjugate CD74-L5-P1, isotype IgG-L5-P1 ADC, and MCL-1 free payload P1 alone or in combination with venetoclax against MONO-MAC-1, NOMO-1, and EOL-1. [Figure 5] FIG. 1 shows the in vitro activity of CD74 MCL-1 antibody drug conjugate CD74-L5-P1, isotype IgG-L5-P1 ADC, and MCL-1 free payload P1 alone or in combination with compound A1 against MONO-MAC-1, NOMO-1, and EOL-1. [Figure 6] Figure 1 shows tumor volume (mm3) in female SCID mice implanted with Nomo1 upon treatment with IgG1-linker-payload Fcsilent, anti-CD74_CysmAb Fcsilent, and anti-CD74_CysmAb Fcsilent_L5-P1 (30 mg / kg, 1 IV dose) alone or in combination with venetoclax (50 mpk, 4ON / 3OFF / 4ON PO dose) (n=6). Triangles and diamonds indicate venetoclax and antibody treatment schedules, respectively. [Figure 7]Figure 1 shows % weight loss in Nomo1-implanted female SCID mice upon treatment with IgG1-linker-payload Fcsilent, anti-CD74_CysmAb Fcsilent and anti-CD74_CysmAb Fcsilent_L5-P1 (30 mg / kg, 1 IV dose) alone or in combination with venetoclax (50 mpk, 4ON / 3OFF / 4ON PO dose) (n=6). [Figure 8] Figure 1 shows tumor volume (mm3) in female NSG mice implanted with Karpas422 upon treatment with IgG1-linker-payload Fcsilent, anti-CD74_CysmAb Fcsilent, and anti-CD74_CysmAb Fcsilent_L5-P1 (30 mg / kg, 1 IV dose) alone or in combination with venetoclax (50 mpk, 4 ON / 3 OFF / 4 ON PO) (n=6). Triangles and diamonds indicate venetoclax and antibody treatment schedules, respectively. [Figure 9] Figure 1 shows % weight loss in Karpas422-implanted female NSG mice upon treatment with IgG1-linker-payload Fcsilent, anti-CD74_CysmAb Fcsilent and anti-CD74_CysmAb Fcsilent_L5-P1 (30 mg / kg, 1 IV dose) alone or in combination with venetoclax (50 mpk, 4ON / 3OFF / 4ON PO dose) (n=6). [Figure 10] Figure 1 shows tumor volume (mm3) in female SCID mice implanted with Monomac1 upon treatment with IgG1-linker-payload Fcsilent, anti-CD74_CysmAb Fcsilent, and anti-CD74_CysmAb Fcsilent_L5-P1 (30 mg / kg, 1 IV dose) alone or in combination with venetoclax (50 mpk, 2ON / 2OFF / 5ON / 2OFF / 4ON PO dose) (n=6). Triangles and diamonds represent venetoclax and antibody treatment schedules, respectively. [Figure 11]Figure 1 shows % weight loss in Monomac1-implanted female SCID mice upon treatment with IgG1-linker-payload Fcsilent, anti-CD74_CysmAb Fcsilent and anti-CD74_CysmAb Fcsilent_L5-P1 (30 mg / kg, 1 IV dose) alone or in combination with venetoclax (50 mpk, 2ON / 2OFF / 5ON / 2OFF / 4ON PO dose) (n=6). [Figure 12] Figure 1 shows tumor volume (mm3) in female SCID mice xenografted with EOL1 human acute myeloid (eosinophilic) leukemia upon treatment with CD74-L5-P1 Fcsilent in combination with venetoclax, along with various control groups. [Figure 13] Figure 1 shows % weight loss in female SCID mice xenografted with EOL1 human acute myeloid (eosinophilic) leukemia upon treatment with CD74-L5-P1 Fcsilent in combination with venetoclax, along with various control groups. [Figure 14] FIG. 1 shows tumor volume (mm3) in female SCID mice xenografted with EOL1 human acute myeloid (eosinophilic) leukemia upon treatment with CD74-ADC and different linker payloads (L5-P1, L30-P1, L31-P1, L32-P1, L33-P1, and L34-P1). [Figure 15] FIG. 1 shows % weight loss in female SCID mice xenografted with EOL1 human acute myeloid (eosinophilic) leukemia upon treatment with CD74-ADC and different linker payloads (L5-P1, L30-P1, L31-P1, L32-P1, L33-P1, and L34-P1). [Figure 16] FIG. 1 shows an exemplary site-specific antibody conjugation using bacterial transglutaminase (BTG). DETAILED DESCRIPTION OF THE INVENTION

[0275] The disclosed compositions and methods can be more readily understood by reference to the following detailed description in conjunction with the accompanying drawings, which form a part of this disclosure.

[0276] Throughout this text, the description refers to compositions and methods of using the compositions. Where the disclosure describes or claims features or embodiments related to compositions, such features or embodiments are equally applicable to methods of using the compositions. Similarly, where the disclosure describes or claims features or embodiments related to methods of using the compositions, such features or embodiments are equally applicable to the compositions.

[0277] When a range of values ​​is expressed, this includes embodiments using any specific value within the range. Furthermore, reference to values ​​stated in a range includes each and every value within that range. All ranges are inclusive of their endpoints and are combinable. Similarly, when values ​​are expressed as approximations, by use of the antecedent "about," it is understood that the particular value forms another embodiment. Reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise. The use of "or" is intended to mean "and / or" unless the particular context of its use dictates otherwise. All references cited herein are incorporated by reference for any purpose. In the event of a conflict between a reference and the present specification, the present specification will control.

[0278] Unless otherwise indicated in the context of this specification, when a structure or fragment of a structure is depicted, e.g., without a symbol indicating a particular point of attachment, it can be used by itself or attached to other components of an ADC, in any orientation, e.g., by attaching an antibody to a chemical moiety, e.g., a linker-drug, at any suitable point of attachment. However, when shown, the components of the ADC are attached in the orientation shown in a given formula. For example, if formula (1) is Ab-(LD) p The group "-(LD)" is

[0279] [ka] When explained as, the compact structure of Eq.

[0280] [ka] This is

[0281] [ka] isn't it.

[0282] It will be appreciated that certain features of the disclosed compositions and methods that are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed compositions and methods that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0283] As used throughout this application, antibody drug conjugates can be identified using a naming convention of the general form "target antigen / antibody-linker-payload." By way of example only, if an antibody drug conjugate is referred to as "target X-L0-P0," such a conjugate would comprise an antibody that binds to target X, a linker designated L0, and a payload designated P0. Alternatively, if an antibody drug conjugate is referred to as "anti-target X-L0-P0," such a conjugate would comprise an antibody that binds to target X, a linker designated L0, and a payload designated P0. Alternatively, if an antibody drug conjugate is referred to as "AbX-L0-P0," such a conjugate would comprise an antibody designated AbX, a linker designated L0, and a payload designated P0. A control antibody drug conjugate comprising a nonspecific isotype control antibody may be referred to as "isotype control IgG1-L0-P0" or "IgG1-L0-P0."

[0284] Any formula given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures represented by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Isotopes that can be incorporated into the compounds of the invention include, for example, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, and chlorine, e.g., 3 H, 11 C. 13 C. 14 C. 15 N, 18 F and 36 Cl. Thus, the present disclosure provides, for example, radioisotopes, e.g. 3 H and 14 Compounds incorporating one or more of any of the above-mentioned isotopes, including C, or non-radioactive isotopes therein, e.g. 2 H and 13 It should be understood that this includes compounds in which C is present. Such isotope-labeled compounds may be used in metabolic studies ( 14 C), reaction kinetics studies (e.g. 2 H or 3 H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including tissue distribution assays of drugs or substances, or in radiation treatment of patients. 18 F or labeled compounds may be particularly desirable for PET or SPECT studies. Isotopically labeled compounds can generally be prepared by conventional techniques known to those of skill in the art, e.g., by substituting an appropriate isotopically labeled reagent for a previously employed unlabeled reagent.

[0285] definition Various terms relating to the described embodiments are used throughout the specification and claims. Such terms will be given their ordinary meaning in the art unless otherwise specified. Other specifically defined terms will be interpreted in a manner consistent with the definitions provided herein.

[0286] As used herein, the singular forms "a," "an," and "the" include the plural forms unless the context dictates otherwise. The terms "comprising," "having," "being of the formula," "including," and "containing" are to be construed as open terms (i.e., meaning "including, but not limited to") unless otherwise noted. In addition, whenever "comprising" or another open-ended term is used in an embodiment, it is understood that the same embodiment can also be more narrowly claimed using the introductory term "consisting essentially of" or the closed term "consisting of."

[0287] The terms "about" or "approximately," when used in the context of numerical values ​​and ranges, refer to a value or range that is close to or near the recited value or range, such that embodiments can be performed as intended, as would be apparent to one of ordinary skill in the art from the teachings contained herein. In some embodiments, about means plus or minus 20%, 15%, 10%, 5%, 1%, 0.5%, or 0.1% of the quantity. In one embodiment, the term "about" refers to a range of values ​​that is 10% above or below a particular value. In another embodiment, the term "about" refers to a range of values ​​that is 5% above or below a particular value. In another embodiment, the term "about" refers to a range of values ​​that is 1% above or below a particular value.

[0288] The terms "antibody-drug conjugate," "antibody conjugate," "conjugate," "immunoconjugate," and "ADC" are used interchangeably and refer to one or more therapeutic compounds (e.g., Mcl-1 inhibitors) linked to one or more antibodies or antigen-binding fragments. In some embodiments, an ADC has the general formula: Ab-(LD) pADCs are defined by Formula 1, where Ab = antibody or antigen-binding fragment, L = linker moiety, D = drug moiety (e.g., Mcl-1 inhibitor drug moiety), and p = number of drug moieties per antibody or antigen-binding fragment. For ADCs that include Mcl-1 inhibitor drug moieties, "p" refers to the number of Mcl-1 inhibitor compounds linked to the antibody or antigen-binding fragment.

[0289] The term "antibody" in its broadest sense refers to an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, carbohydrate, polynucleotide, lipid, or a combination of the above, through at least one antigen recognition site in the variable region of the immunoglobulin molecule. Antibodies can be polyclonal or monoclonal, multi-chain or single-chain, or intact immunoglobulins, and can be derived from natural or recombinant sources. "Intact" antibodies are typically glycoproteins comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain is composed of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region contains three domains, CH1, CH2, and CH3. Each light chain is composed of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is composed of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability termed complementarity-determining regions (CDRs), interspersed with more conserved regions termed framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino to carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of antibodies can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The antibodies can be monoclonal, human, humanized, camelized, or chimeric. The antibody can be of any isotype (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. The antibody can be an intact antibody or an antigen-binding fragment thereof.

[0290] The terms "antibody fragment" or "antigen-binding fragment" or "functional antibody fragment," as used herein, refer to at least one portion of an antibody that retains the ability to specifically interact (e.g., by binding, steric hindrance, stabilization / destabilization, spatial distribution) with an epitope of an antigen (e.g., CD74). An antigen-binding fragment may also retain the ability to be internalized into antigen-expressing cells. In some embodiments, an antigen-binding fragment also retains immune effector activity. The terms antibody, antibody fragment, antigen-binding fragment, and the like are intended to encompass the use of antibody-derived binding domains in the context of larger macromolecules, such as ADCs. Fragments of a full-length antibody have been shown to be capable of performing the antigen-binding function of the full-length antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab'), Fv fragments, scFv antibody fragments, disulfide-linked Fvs (sdFv), Fd fragments consisting of the VH and CH1 domains, linear antibodies, single-domain antibodies such as sdAb (either VL or VH), camelid VHH domains, antibody fragments, such as multispecific antibodies formed from a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region, and isolated CDRs or other epitope-binding fragments of antibodies. Antigen-binding fragments can also be incorporated into single-domain antibodies, maxibodies, minibodies, nanobodies, intrabodies, diabodies, triabodies, tetrabodies, bispecific or multispecific antibody constructs, ADCs, v-NARs, and bis-scFvs (see, e.g., Holliger and Hudson (2005) Nat Biotechnol. 23(9):1126-36). Antigen-binding fragments can also be grafted into polypeptide-based scaffolds, such as fibronectin type III (Fn3) (see U.S. Pat. No. 6,703,199, which describes fibronectin polypeptide minibodies).The term "scFv" refers to a fusion protein comprising at least one antigen-binding fragment comprising a light chain variable region and at least one antigen-binding fragment comprising a heavy chain variable region, wherein the light and heavy chain variable regions are contiguously linked, e.g., via a synthetic linker, e.g., a short, flexible polypeptide linker, and can be expressed as a single-chain polypeptide, wherein the scFv retains the specificity of the intact antibody from which it is derived. Unless specified, an scFv can have the VL and VH variable regions in either order; e.g., relative to the N- and C-termini of the polypeptide, an scFv can comprise VL-linker-VH or VH-linker-VL. Antigen-binding fragments are obtained using conventional techniques known to those skilled in the art, and binding fragments are screened for utility (e.g., binding affinity, internalization) in the same manner as intact antibodies. Antigen-binding fragments can be prepared, for example, by cleavage of the intact protein, e.g., by protease or chemical cleavage.

[0291] The term "complementarity determining region" or "CDR" as used herein refers to the sequence of amino acids in an antibody variable region that confers antigen specificity and binding affinity. For example, there are generally three CDRs (e.g., HCDR1, HCDR2, and HCDR3) in each heavy chain variable region, and three CDRs (LCDR1, LCDR2, and LCDR3) in each light chain variable region. The precise amino acid sequence boundaries of a given CDR can be determined using any one of a number of well-known schemes, or a combination thereof, including those described by Kabat et al. (1991) "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme); Al-Lazikani et al. (1997) J Mol Biol. 273(4):927-48 ("Chothia" numbering scheme); ImMunoGenTics (IMGT) numbering (Lefranc (2001) Nucleic Acids Res. 29(1):207-9; Lefranc et al. (2003) Dev Comp Immunol. 27(1):55-77) ("IMGT" numbering scheme). In the combined Kabat and Chothia numbering scheme for a given CDR region (e.g., HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, or LC CDR3), in some embodiments, the CDRs correspond to the amino acid residues defined as part of the Kabat CDRs along with the amino acid residues defined as part of the Chothia CDRs. As used herein, CDRs defined by the "Chothia" numbering scheme are sometimes referred to as "hypervariable loops."

[0292] In some embodiments, under Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1) (e.g., insertion after position 35), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1) (e.g., insertion after position 27), 50-56 (LCDR2), and 89-97 (LCDR3). In some embodiments, under Chothia, the CDR amino acids in the VH are numbered 26-32 (HCDR1) (e.g., insertion after position 31), 52-56 (HCDR2), and 95-102 (HCDR3); and the amino acid residues in the VL are numbered 26-32 (LCDR1) (e.g., insertion after position 30), 50-52 (LCDR2), and 91-96 (LCDR3). Combining the CDR definitions of both Kabat and Chothia, in some embodiments, a CDR comprises or consists of, for example, amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in a human VH and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in a human VL. In some embodiments, under IMGT, the CDR amino acid residues in a VH are numbered approximately 26-35 (CDR1), 51-57 (CDR2), and 93-102 (CDR3), and the CDR amino acid residues in a VL are numbered approximately 27-32 (CDR1), 50-52 (CDR2), and 89-97 (CDR3). In some embodiments, under IMGT, the CDR regions of an antibody can be determined using the program IMGT / Domain Gap Align.

[0293] The term "monoclonal antibody," as used herein, 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 minor naturally occurring mutations. Monoclonal antibodies are highly specific, being directed against a single antigenic epitope. In contrast, conventional (polyclonal) antibody preparations typically include many antibodies directed against (or specific for) different epitopes. The modifier "monoclonal" refers to the character of the antibody as being obtained from a population of substantially homogeneous 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 can be made, e.g., by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or can 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, for example, the techniques described in Clackson et al. (1991) Nature 352:624-8 and Marks et al. (1991) J Mol Biol. 222:581-97. The term also includes preparations of antibody molecules of single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope.

[0294] The monoclonal antibodies described herein can be non-human, human, or humanized. The term specifically includes "chimeric" antibodies in which portions of the heavy and / or light chains are identical to or homologous to sequences in corresponding antibodies from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chains are identical to or homologous to corresponding sequences in antibodies from another species or belonging to another antibody class or subclass, and in fragments of such antibodies, so long as they specifically bind to the target antigen and / or exhibit the desired biological activity.

[0295] The term "human antibody," as used herein, refers to an antibody produced by a human or an antibody having the amino acid sequence of an antibody produced by a human. This term includes antibodies having variable regions in which both the framework and CDR regions are derived from sequences of human origin. Furthermore, if the antibody contains a constant region, the constant region is also derived from such a human sequence, e.g., a human germline sequence, or a mutated version of a human germline sequence, or an antibody containing a consensus framework sequence derived from human framework sequence analysis as described in Knappik et al. ((2000) J Mol Biol. 296(1):57-86). The structure and position of immunoglobulin variable domains, e.g., CDRs, can be defined using well-known numbering schemes, such as the Kabat numbering scheme, the Chothia numbering scheme, or a combination of Kabat and Chothia, and / or ImMunoGenTics (IMGT) numbering. The human antibodies of the invention may include amino acid residues not encoded by human sequences (e.g., mutations induced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo, or conservative substitutions to facilitate stability or production). However, the term "human antibody," as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of other mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0296] The term "recombinant human antibody," as used herein, refers to a human antibody that is prepared, expressed, produced, or isolated by recombinant means, e.g., an antibody isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom, an antibody isolated from a host cell, e.g., a transfectoma, that has been transformed to express a human antibody, an antibody isolated from a recombinant combinatorial human antibody library, and an antibody prepared, expressed, produced, or isolated by any other means involving splicing all or part of a human immunoglobulin gene sequence into other DNA sequences. Such recombinant human antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies may be subjected to in vitro mutagenesis (or, if animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis) such that the amino acid sequences of the VH and VL regions of the recombinant antibodies, while derived from and related to human germline VH and VL sequences, are sequences that would not naturally occur in the human antibody germline repertoire in vivo.

[0297] The term "chimeric antibody," as used herein, refers to an antibody in which the amino acid sequences of the immunoglobulin molecules are derived from two or more species. In some cases, the variable regions of both the heavy and light chains correspond to the variable regions of an antibody derived from one species having the desired specificity, affinity, and activity, while the constant regions are homologous to antibodies derived from another species (e.g., humans) to minimize immune responses in the latter species.

[0298] As used herein, "humanized antibody" refers to a form of antibody containing sequences from non-human (e.g., murine) and human antibodies. Such antibodies are a type of chimeric antibody, containing minimal sequence derived from non-human immunoglobulin. Generally, a humanized antibody will contain substantially all of at least one, typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the framework (FR) regions are those of a human immunoglobulin sequence. A humanized antibody will also optionally contain at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. Humanized antibodies can be further modified by substitution of residues either in the Fv framework regions and / or substituted non-human residues to improve and optimize antibody specificity, affinity, and / or activity.

[0299] The term "Fc region," as used herein, refers to a polypeptide comprising at least a portion of the CH3, CH2, and hinge region of an antibody constant domain. Optionally, the Fc region can include a CH4 domain present in some antibody classes. The Fc region can also include the entire hinge region of an antibody constant domain. In some embodiments, an antibody or antigen-binding fragment comprises the Fc region and CH1 region of an antibody. In some embodiments, an antibody or antigen-binding fragment comprises the Fc region and CH3 region of an antibody. In some embodiments, an antibody or antigen-binding fragment comprises the Fc region, CH1 region, and kappa / lambda regions from the antibody constant domain. In some embodiments, an antibody or antigen-binding fragment comprises a constant region, e.g., a heavy chain constant region and / or a light chain constant region. In some embodiments, such constant regions are modified compared to wild-type constant regions. That is, the polypeptide can include changes or modifications to one or more of the three heavy chain constant domains (CH1, CH2, or CH3) and / or light chain constant region domain (CL). Examples of modifications include the addition, deletion, or substitution of one or more amino acids in one or more domains. Such changes may be included to optimize effector function, half-life, etc.

[0300] "Internalization," as used herein in reference to an antibody or antigen-binding fragment, refers to an antibody or antigen-binding fragment that, upon binding to a cell, is capable of passing through the lipid bilayer membrane of the cell and being taken up (i.e., "internalized") into an internal compartment, preferably a degradable compartment within the cell. For example, an anti-HER2 antibody is internalized such that it is capable of being taken up into the cell after binding to HER2 on the cell membrane. In some embodiments, the antibody or antigen-binding fragment used in the ADCs disclosed herein targets a cell surface antigen (e.g., CD74) and is an internalizing antibody or internalizing antigen-binding fragment (i.e., the ADC translocates through the cell membrane after antigen binding). In some embodiments, the internalizing antibody or antigen-binding fragment binds to a receptor on the cell surface. An internalizing antibody or internalizing antigen-binding fragment that targets a receptor on the cell membrane can induce receptor-mediated endocytosis. In some embodiments, the internalizing antibody or internalizing antigen-binding fragment is taken up into the cell via receptor-mediated endocytosis.

[0301] "Non-internalizing," as used herein in reference to an antibody or antigen-binding fragment, refers to an antibody or antigen-binding fragment that remains on the cell surface upon binding to a cell. In some embodiments, the antibody or antigen-binding fragment used in the ADCs disclosed herein targets a cell surface antigen and is a non-internalizing antibody or non-internalizing antigen-binding fragment (i.e., the ADC remains on the cell surface after antigen binding and does not translocate through the cell membrane). In some embodiments, the non-internalizing antibody or antigen-binding fragment binds to a non-internalizing receptor or other cell surface antigen. Representative non-internalizing cell surface antigens include, but are not limited to, CA125 and CEA, and antibodies that bind to non-internalizing antigen targets are also known in the art (see, e.g., Bast et al. (1981) J Clin Invest. 68(5):1331-7; Scholler and Urban (2007) Biomark Med. 1(4):513-23; and Boudousq et al. (2013) PLoS One 8(7):e69613).

[0302] The term "binding specificity," as used herein, refers to the ability of an individual antibody or antigen-binding fragment to preferentially react with one antigenic determinant over different antigenic determinants. Specificity refers to the degree to which an antibody or fragment preferentially binds to one antigenic determinant over different antigenic determinants. Also, as used herein, the terms "specific," "specifically binds," and "binds specifically" refer to the binding reaction between an antibody or antigen-binding fragment (e.g., an anti-CD74 antibody) and a target antigen (e.g., CD74) in a heterogeneous population of proteins and other biologics. An antibody can be tested for binding specificity by comparing binding to the appropriate antigen with binding to an unrelated antigen or antigen mixture under a given set of conditions. An antibody is considered specific if it binds to the appropriate antigen with at least 2-fold, 5-fold, 7-fold, or 10-fold greater affinity than an unrelated antigen or antigen mixture. A "specific antibody" or "target-specific antibody" is one that binds only to a target antigen (e.g., CD74) and does not bind (or shows minimal binding to) other antigens. In some embodiments, an antibody or antigen-binding fragment that specifically binds to a target antigen (e.g., CD74) is at least 1x10 -6 Under M, 1x10 -7 Under M, 1x10 -8 Under M, 1x10 -9 Under M, 1x10 -10 Under M, 1x10 -11 Under M, 1x10 -12 M or 1x10 -13 K less than M D In some embodiments, K D In some embodiments, K D is between 500 pM and 1 μM, 1 μM and 100 nM, or 100 mM and 10 nM.

[0303] The term "affinity," as used herein, refers to the strength of the interaction between an antibody and an antigen at a single antigenic site. Without being bound by theory, within each antigen-binding site, the antibody variable region "arms" interact with the antigen at numerous sites through weak non-covalent forces, and the more interactions there are, the stronger the affinity typically becomes. The binding affinity of an antibody is the sum of the attractive and repulsive forces acting between an antigenic determinant and the antibody binding site.

[0304] "k on " or "k a The term "on-rate constant" refers to the on-rate constant for the association of an antibody to an antigen to form an antibody / antigen complex. This rate can be determined using standard assays, such as surface plasmon resonance, biolayer interferometry, or ELISA assays.

[0305] "k off " or "k d The term "off-rate constant" refers to the off-rate constant for dissociation of an antibody from the antibody / antigen complex. This rate can be determined using standard assays, such as surface plasmon resonance, biolayer interferometry, or ELISA assays.

[0306] "K D The term K refers to the equilibrium dissociation constant of a particular antibody-antigen interaction. D is k a / k d This rate can be determined using standard assays, such as surface plasmon resonance, biolayer interferometry, or ELISA assays.

[0307] The term "epitope" refers to that portion of an antigen capable of being recognized and specifically bound by an antibody (or antigen-binding fragment). Epitopic determinants generally consist of chemically active surface groupings of molecules such as amino acids or carbohydrate or sugar side chains and can have specific three-dimensional structural characteristics, as well as specific charge characteristics. When the antigen is a polypeptide, epitopes can be formed from contiguous amino acids or noncontiguous amino acids juxtaposed by tertiary folding of the polypeptide. Epitopes can be "linear" or "conformational." Conformational epitopes are distinguished from linear epitopes in that the binding to the former but not the latter is lost in the presence of denaturing solvents. The epitope bound by an antibody (or antigen-binding fragment) can be identified using any epitope mapping technique known in the art, including X-ray crystallography for epitope identification by direct visualization of the antigen-antibody complex, and monitoring binding of the antibody to fragments or mutated versions of the antigen, or monitoring the solvent accessibility of different portions of the antibody and antigen. Exemplary strategies for mapping antibody epitopes include, but are not limited to, array-based oligo-peptide scanning, limited proteolysis, site-directed mutagenesis, high-throughput mutagenesis mapping, hydrogen-deuterium exchange, and mass spectrometry (see, e.g., Gershoni et al. (2007) BioDrugs 21:145-56; and Hager-Braun and Tomer (2005) Expert Rev Proteomics 2:745-56).

[0308] Competitive binding and epitope binning can also be used to determine antibodies that share the same or overlapping epitopes. Competitive binding can be assessed using a cross-blocking assay, such as the assay described in "Antibodies, A Laboratory Manual," Cold Spring Harbor Laboratory, Harlow and Lane (1st edition 1988, 2nd edition 2014). In some embodiments, competitive binding is identified when a test antibody or binding protein reduces the binding of a reference antibody or binding protein (e.g., a binding protein comprising a CDR and / or variable domain selected from those identified in Tables 3-5) to a target antigen, e.g., CD74, by at least about 50% (e.g., 50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.5% or more, or any percentage therebetween) in a cross-blocking assay, and / or vice versa. In some embodiments, competitive binding can be due to shared or similar (e.g., particularly overlapping) epitopes, or due to steric hindrance of antibodies or binding proteins binding to nearby epitopes (see, e.g., Tzartos, Methods in Molecular Biology (Morris, ed. (1998) vol. 66, pp. 55-66)). In some embodiments, competitive binding can be used to sort groups of binding proteins that share similar epitopes. For example, binding proteins that compete for binding can be "binned" into a group of binding proteins with overlapping or nearby epitopes, while those that do not compete are placed into a separate group of binding proteins that do not have overlapping or nearby epitopes.

[0309] As used herein, the terms "peptide," "polypeptide," and "protein" are used interchangeably and refer to a polymer of amino acid residues. The term encompasses amino acid polymers containing two or more amino acids connected together by peptide bonds, amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of corresponding naturally occurring amino acids, and naturally occurring and non-naturally occurring amino acid polymers. The term specifically encompasses, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, polypeptide variants, modified polypeptides, derivatives, analogs, and fusion proteins. The term also encompasses natural peptides, recombinant peptides, synthetic peptides, or combinations thereof. Unless otherwise specified, a particular polypeptide sequence implicitly encompasses conservatively modified variants thereof.

[0310] A "recombinant" protein refers to a protein (eg, an antibody) made using recombinant techniques, eg, through the expression of a recombinant nucleic acid.

[0311] An "isolated" protein refers to a protein that is free from at least some of the materials that normally accompany it in its natural state. For example, a naturally occurring polynucleotide or polypeptide present in a living organism is not isolated, but the same polynucleotide or polypeptide separated from some or all of the coexisting materials in a living organism is isolated. This definition includes the production of antibodies in a wide variety of organisms and / or host cells known in the art.

[0312] An "isolated antibody," as used herein, is an antibody that has been identified and separated from one or more (e.g., multiple) components of its source environment, such as components of a hybridoma cell culture or a different cell culture used to produce it. In some embodiments, separation is performed to sufficiently remove components that may otherwise interfere with the suitability of the antibody for a desired use (e.g., therapeutic use). Methods for preparing isolated antibodies are known in the art and include, without limitation, protein A chromatography, anion exchange chromatography, cation exchange chromatography, virus retentive filtration, and ultrafiltration.

[0313] As used herein, the term "variant" refers to a nucleic acid sequence or amino acid sequence that differs from a reference nucleic acid sequence or amino acid sequence, respectively, but retains one or more biological properties of the reference sequence. A variant can contain one or more amino acid substitutions, deletions, and / or insertions (or corresponding codon substitutions, deletions, and / or insertions) relative to the reference sequence. Changes in a nucleic acid variant may not change the amino acid sequence of a peptide encoded by the reference nucleic acid sequence, or may result in amino acid substitutions, additions, deletions, fusions, and / or truncations. In some embodiments, the nucleic acid variants disclosed herein encode an amino acid sequence identical to that encoded by the unmodified nucleic acid, or encode a modified amino acid sequence that retains one or more functional properties of the unmodified amino acid sequence. Changes in the sequence of a peptide variant are typically limited or conservative, so that the sequences of the unmodified peptide and the variant are closely similar overall and, in many regions, identical. In some embodiments, a peptide variant retains one or more functional properties of the unmodified peptide sequence. A variant may differ in amino acid sequence from the unmodified peptide by one or more substitutions, additions, deletions in any combination.

[0314] Nucleic acid or peptide variants can be naturally occurring variants or variants that are not known to occur naturally.Nucleic acid and peptide variants can be produced by mutagenesis techniques, by direct synthesis, or by other techniques known in the art.Variant does not necessarily require physical manipulation of the reference sequence.As long as a sequence contains different nucleic acids or amino acids compared to the reference sequence, it is considered a "variant" regardless of how much it is synthesized.In some embodiments, variants have high sequence identity (i.e., 60% or more identity of nucleic acid or amino acid sequence) compared to the reference sequence. In some embodiments, peptide variants include polypeptides with amino acid substitutions, deletions, and / or insertions, so long as the polypeptide has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% amino acid sequence identity with a reference sequence or a corresponding segment (e.g., functional fragment) of a reference sequence, e.g., a variant that also retains one or more functions of the reference sequence. In some embodiments, nucleic acid variants include polynucleotides with amino acid substitutions, deletions, and / or insertions, so long as the polynucleotide has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% nucleic acid sequence identity to a reference sequence or a corresponding segment (e.g., a functional fragment) of a reference sequence.

[0315] The term "conservatively modified variants" applies to both amino acid and nucleic acid sequences. For nucleic acid sequences, conservatively modified variants refer to nucleic acids that encode identical or essentially identical amino acid sequences. Due to the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For example, the codons GCA, GCC, GCG, and GCU all encode the amino acid alanine. Thus, at every position where alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," which are a type of conservatively modified variation. Every nucleic acid sequence herein that encodes a polypeptide also describes all possible silent variations of the nucleic acid. Those skilled in the art will recognize that each codon in a nucleic acid (except AUG, the only codon for conventional methionine, and TGG, the only codon for conventional tryptophan) can be modified to obtain a functionally identical molecule. Thus, each silent variation of a nucleic acid that encodes a polypeptide is implicit in each described sequence. For polypeptide sequences, conservatively modified variants include individual substitutions, deletions, or additions to the polypeptide sequence that result in the substitution of an amino acid with a chemically similar amino acid. Conservative substitutions that provide functionally similar amino acids are well known in the art.

[0316] The term "conservative sequence modification" as used herein refers to an amino acid modification that does not significantly affect or alter the binding properties of, for example, an antibody or antigen-binding fragment containing the amino acid sequence. Such conservative modifications include amino acid substitutions, additions, and deletions. Modifications can be introduced into antibodies or antigen-binding fragments by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. A conservative amino acid substitution is one in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, in some embodiments, one or more amino acid residues in an antibody can be substituted with other amino acid residues from the same side chain family, and the altered antibodies can be tested using the functional assays described herein.

[0317] The term "homologous" or "identity," as used herein, refers to the sequence identity of subunits between two polymer molecules, e.g., between two nucleic acid molecules, e.g., two DNA molecules or two RNA molecules, or between two polypeptide molecules. If a subunit position in both of the two molecules is occupied by the same monomer subunit, e.g., if each position in two DNA molecules is occupied by adenine, they are homologous or identical at that position. The homology between two sequences is a linear function of the number of matching or homologous positions. For example, if half of the positions in two sequences (e.g., five positions in a polymer 10 subunits long) are matched or homologous, the two sequences are 50% homologous; if 90% of the positions (e.g., 9 out of 10) are matched or homologous, the two sequences are 90% homologous.

[0318] The percentage of "sequence identity" can be determined by comparing two optimally aligned sequences over a comparison window, where the amino acid sequence fragment in the comparison window can contain additions or deletions (e.g., gaps or overhangs) compared to the reference sequence (which contains no additions or deletions) for optimal alignment of the two sequences. The percentage can be calculated by determining the number of positions where identical amino acid residues occur in both sequences to obtain the number of matching positions, dividing the number of matching positions by the total number of positions in the comparison window, and multiplying the result by 100 to obtain the percentage of sequence identity. The output is the percent identity of the subject sequence to the query sequence. The percent identity between two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps that need to be introduced for optimal alignment of the two sequences and the length of each gap. Generally, the amino acid identity or homology between the proteins disclosed herein and variants thereof, including variants of target antigens (e.g., CD74) and variants of antibody variable domains (including individual variant CDRs), is at least 80% of the sequences set forth herein, such as at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, nearly 100% or 100% identity or homology.

[0319] Comparison of sequences and determination of percent identity between two sequences can be accomplished using a numerical algorithm. In some embodiments, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch ((1970) J Mol Biol. 48:444-53) algorithm incorporated into the GAP program in the GCG software package, using either a Blossum62 matrix or a PAM250 matrix, a gap weight of 16, 14, 12, 10, 8, 6, or 4, and a length weight of 1, 2, 3, 4, 5, or 6. In some embodiments, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package, using a NWSgapdna.CMP matrix, a gap weight of 40, 50, 60, 70, or 80, and a length weight of 1, 2, 3, 4, 5, or 6. A representative set of parameters is a Blossum62 score matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5. The percent identity between two amino acid or nucleotide sequences may also be determined using the algorithm of Meyers and Miller ((1989) CABIOS 4:11-17) as incorporated into the ALIGN program (version 2.0), using the PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.

[0320] The term "drug" is used herein to refer to a chemical compound, a mixture of chemical compounds, a biological macromolecule, an extract made from biological materials, or a combination of two or more thereof. The term "therapeutic agent" or "drug" refers to an agent capable of modulating biological processes and / or biological activity. Mcl-1 inhibitors as described herein and ADCs containing them are representative therapeutic agents.

[0321] The term "chemotherapeutic agent" or "anti-cancer agent" is used herein to refer to all agents effective in treating cancer (regardless of mechanism of action). Inhibition of metastasis or angiogenesis is often a property of chemotherapeutic agents. Chemotherapeutic agents include antibodies, biological molecules, and small molecules, including Mcl-1 inhibitors as described herein and ADCs containing them. Chemotherapeutic agents may be cytotoxic or cytostatic agents. The term "cytostatic agent" refers to an agent that inhibits or suppresses cell growth and / or cell proliferation. The term "cytotoxic agent" refers to a substance that causes cell death primarily by interfering with cellular expression activity and / or function.

[0322] The term "myeloid cell leukemia 1" or "Mcl-1," as used herein, refers to any naturally occurring form of human Mcl-1, an anti-apoptotic member of the Bcl-2 protein family. This term encompasses full-length human Mcl-1 (e.g., UniProt Reference Sequence: Q07820; SEQ ID NO: 63) and any form of human Mcl-1 that may result from cellular processing. This term also encompasses functional variants or fragments of human Mcl-1, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human Mcl-1 (i.e., variants and fragments are included unless the context indicates that the term is used to refer only to the wild-type protein). Mcl-1 can be isolated from humans or produced recombinantly or by synthetic methods.

[0323] The term "inhibit" or "inhibition" or "inhibiting," as used herein, means to reduce a biological activity or process by a measurable amount, but may include, but does not require, complete prevention or inhibition. In some embodiments, "inhibition" refers to reducing the expression and / or activity of Mcl-1 and / or one or more of its upstream regulators or downstream targets.

[0324] The term "Mcl-1 inhibitor," as used herein, refers to an agent capable of reducing the expression and / or activity of Mcl-1 and / or one or more of its upstream regulators or downstream targets. Representative Mcl-1 regulators (including representative inhibitors of Mcl-1) are described in WO 2015 / 097123; WO 2016 / 207216; WO 2016 / 207217; WO 2016 / 207225; WO 2016 / 207226; WO 2017 / 125224; WO 2019 / 035899; WO 2019 / 035911; WO 2019 / 035914.

[0004] The present invention relates to a method for the preparation of Mcl-1 modulators, such as those described in FRET, WO 2019 / 035927, U.S. Patent Application Publication No. 2019 / 0055264, WO 2016 / 033486, WO 2017 / 147410, WO 2018 / 183418, and WO 2017 / 182625, each of which is incorporated by reference herein as representative Mcl-1 modulators, including representative Mcl-1 inhibitors that can be included as drug moieties in the disclosed ADCs. For example, a representative Mcl-1 inhibitor that can be included as a drug moiety in the disclosed ADCs has the formula:

[0325] [ka] wherein each variable is defined as in WO 2019 / 035911; WO 2019 / 035899; WO 2019 / 035914; or WO 2019 / 035927. Specific examples include, for example:

[0326] [ka] wherein each compound as a drug payload can be conjugated to an antibody or a linker via the N-methyl nitrogen atom in the piperazinyl functional group of the compound. As used herein, the terms "derivative" and "analog," when referring to Mcl-1 inhibitors and the like, refer to any such compound that retains essentially the same, similar, or enhanced biological function or activity compared to the original compound, but has an altered chemical or biological structure.

[0327] As used herein, "Mcl-1 inhibitor drug moiety," "Mcl-1 inhibitor," and the like refer to a component of an ADC or composition that provides a compound structure modified for attachment to an ADC that retains essentially the same, similar, or enhanced biological function or activity compared to the Mcl-1 inhibitor compound or the parent compound. In some embodiments, the Mcl-1 inhibitor drug moiety is component (D) in an ADC of Formula (1).

[0328] The term "cancer," as used herein, refers to the presence of cells with characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and / or certain morphological characteristics. While cancer cells often take the form of a tumor or mass, such cells may exist alone in a subject or circulate in the bloodstream as individual cells, e.g., leukemia or lymphoma cells. The term "cancer" includes all types of cancer and cancer metastasis, including blood cancers, solid tumors, sarcomas, carcinomas, and other solid and non-solid tumor cancers. Blood cancers can include B-cell malignancies, cancers of the blood (leukemia), cancers of plasma cells (myeloma, e.g., multiple myeloma), or cancers of the lymph nodes (lymphoma). Representative B-cell malignancies include chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, and diffuse large B-cell lymphoma. Leukemias include acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), etc. Lymphomas include Hodgkin's lymphoma, non-Hodgkin's lymphoma, etc. Other blood cancers include myelodysplastic syndromes (MDS). Solid tumors include carcinomas, such as adenocarcinomas, e.g., breast cancer, pancreatic cancer, prostate cancer, colon or colorectal cancer, lung cancer, gastric cancer, cervical cancer, endometrial cancer, ovarian cancer, cholangiocarcinoma, glioma, melanoma, etc. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[0329] In some embodiments, the cancer is a blood cancer, such as leukemia, lymphoma or myeloma.For example, the combinations described herein can be used in, but are not limited to, acute leukemia, such as B-cell acute lymphoblastic leukemia (BALL), T-cell acute lymphoblastic leukemia (TALL), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL); chronic leukemia, such as chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL); additional blood cancers or blood conditions, such as B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell or large cell follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, It can be used to treat cancer malignancies and related disorders, including mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndromes, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom's macroglobulinemia, myelofibrosis, amyloid light chain amyloidosis, chronic neutrophilic leukemia, essential thrombocythemia, chronic eosinophilic leukemia, chronic myelomonocytic leukemia, Richter's syndrome, mixed phenotype acute leukemia, acute mixed phenotype leukemia, and "preleukemias," a diverse collection of blood conditions united by ineffective production (or dysplasia) of bone marrow blood cells.

[0330] As used herein, the term "tumor" refers to any mass of tissue resulting from excessive cell growth or proliferation, whether benign or malignant, including precancerous lesions. In some embodiments, the tumor is breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular carcinoma, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the tumor is gastric cancer.

[0331] The terms "tumor cell" and "cancer cell" can be used interchangeably herein and refer to an individual cell or the entire population of cells derived from a tumor or cancer, including both non-tumorigenic cells and cancer stem cells. When referring only to cells that have lost the ability to reproduce and differentiate, the terms "tumor cell" and "cancer cell" will be modified by the term "non-tumorigenic" to distinguish these cells from cancer stem cells.

[0332] The terms "target negative," "target antigen negative," or "antigen negative," as used herein, refer to the absence of target antigen expression by a cell or tissue. The terms "target positive," "target antigen positive," or "antigen positive" refer to the presence of target antigen expression. For example, a cell or cell line that does not express a target antigen can be described as target negative, while a cell or cell line that expresses a target antigen can be described as target positive.

[0333] The terms "subject" and "patient" are used interchangeably herein and refer to any human or non-human animal in need of treatment. Non-human animals include all vertebrates (e.g., mammals and non-mammals), such as any mammal. Non-limiting examples of mammals include humans, chimpanzees, apes, monkeys, cows, horses, sheep, goats, pigs, rabbits, dogs, cats, rats, mice, and guinea pigs. Non-limiting examples of non-mammals include birds and fish. In some embodiments, the subject is a human.

[0334] The term "subject in need of treatment," as used herein, refers to a subject who would benefit biologically, medically, or in quality of life from treatment (e.g., treatment with any one or more of the representative ADC compounds described herein).

[0335] As used herein, the terms "treat," "treating," or "treatment" refer to any improvement in any outcome of a disease, disorder, or condition, e.g., extending survival, reducing morbidity, and / or reducing side effects resulting from alternative therapies. In some embodiments, treatment includes delaying or reversing the disease, disorder, or condition (i.e., delaying, halting, or reducing the onset of the disease or at least one of its clinical symptoms). In some embodiments, treatment includes delaying, alleviating, or reversing at least one physical parameter of the disease, disorder, or condition, including those that may not be discernible by the patient. In some embodiments, treatment includes modulating the disease, disorder, or condition either physically (e.g., stabilizing discernible symptoms), physiologically (e.g., stabilizing physical parameters), or both. In some embodiments, treatment includes administering a described ADC compound or composition to a subject, e.g., a patient, to obtain the treatment benefits enumerated herein. Treatment can be to cure, ameliorate, alleviate, delay, prevent, relieve, alter, counteract, ameliorate, ameliorate, or affect a disease, disorder, or condition (e.g., cancer), a symptom of a disease, disorder, or condition (e.g., cancer), or the propensity for a disease, disorder, or condition (e.g., cancer). In some embodiments, in addition to treating a subject with a disease, disorder, or condition, the compositions disclosed herein can also be provided prophylactically to prevent or reduce the likelihood of developing the disease, disorder, or condition.

[0336] As used herein, the terms "prevent," "preventing," or "prevention" of a disease, disorder, or condition refers to prophylactic treatment of a disease, disorder, or condition, or delaying the onset or progression of a disease, disorder, or condition.

[0337] As used herein, a "pharmaceutical composition" refers to a composition, e.g., a formulation of an ADC compound or composition, in addition to at least one other component (and optionally more than one other component), e.g., a pharmaceutically acceptable carrier, stabilizer, diluent, dispersant, suspending agent, thickener, and / or excipient, that is suitable for administration to a subject. The pharmaceutical compositions provided herein are in a form that allows for administration and subsequently provides the intended biological activity of the active ingredient and / or achieves a therapeutic effect. The pharmaceutical compositions provided herein preferably do not contain additional components that are unacceptably toxic to the subject to whom the formulation will be administered.

[0338] As used herein, the terms "pharmaceutically acceptable carrier" and "physiologically acceptable carrier" can be used interchangeably and refer to a carrier or diluent that does not cause significant irritation to a subject and does not abrogate the biological activity and properties of the administered ADC compound or composition and / or any additional therapeutic agents in the composition. Pharmaceutically acceptable carriers can enhance or stabilize a composition or can be used to facilitate preparation of the composition. Pharmaceutically acceptable carriers can include solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavorings, dyes, and the like, and combinations thereof, as known to those skilled in the art (see, e.g., Remington's Pharmaceutical Sciences, 18th Ed., Mack Printing Company, 1990, pp. 1289-1329). Unless any conventional carrier is incompatible with active ingredient, its use in therapeutic or pharmaceutical compositions is contemplated.Carrier can be selected to minimize adverse side effects in subject and / or minimize the deterioration of active ingredient.Adjuvants can also be included in any of these formulations.

[0339] As used herein, the term "excipient" refers to an inert substance added to a pharmaceutical composition to facilitate the administration of active ingredients.Formulations for parenteral administration can contain excipients such as sterile water or saline, polyalkylene glycols such as polyethylene glycol, vegetable oils or hydrogenated naphthalene.Other representative excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and starch types, cellulose derivatives, gelatin, ethylene-vinyl acetate copolymer particles, and surfactants, including, for example, polysorbate 20.

[0340] The term "pharmaceutically acceptable salts" as used herein refers to salts that do not abrogate the biological activity and properties of the compounds of the present invention and do not cause significant irritation to the subject to which they are administered. Examples of such salts include, but are not limited to: (a) acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.; and salts formed with organic acids, such as acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, etc.; and (b) salts formed from elemental anions, such as chlorine, bromine, and iodine. See, for example, Haynes et al., "Commentary: Occurrence of Pharmaceutically Acceptable Anions and Cations in the Cambridge Structural Database," J. Pharmaceutical Sciences, vol. 94, no. 10 (2005), and Berge et al., "Pharmaceutical Salts," J. Pharmaceutical Sciences, vol. 66, no. 1 (1977), which are incorporated by reference herein.

[0341] In some embodiments, the antibody-drug conjugates (ADCs), linkers, payloads, and linker-payloads described herein may be coupled to a monovalent anionic counterion M1 depending on their charge. - Any suitable anionic counterion can be used. In certain embodiments, the monovalent anionic counterion is a pharmaceutically acceptable monovalent anionic counterion. In certain embodiments, the monovalent anionic counterion M1 - can be selected from bromide, chloride, iodide, acetate, trifluoroacetate, benzoate, mesylate, tosylate, triflate, formate, etc. In some embodiments, the monovalent anionic counterion M - is a trifluoroacetate or formate.

[0342] As used herein, the term "therapeutically effective amount" or "therapeutically effective dose" refers to the amount of a compound described herein, e.g., an ADC compound or composition described herein, to produce a desired therapeutic result (i.e., reduction or inhibition of enzyme or protein activity, amelioration of symptoms, alleviation of a symptom or condition, delay in disease progression, reduction in tumor size, inhibition of tumor growth, prevention of metastasis). In some embodiments, a therapeutically effective amount does not induce or cause undesirable side effects. In some embodiments, a therapeutically effective amount induces and causes only those side effects that are tolerable by the treating physician in light of the patient's condition. In some embodiments, a therapeutically effective amount is effective to detectably kill, reduce, and / or inhibit the growth or spread of cancer cells, tumor size or number, and / or other measures of the level, stage, progression, and / or severity of cancer. The term also applies to a dose that will induce a specific response in target cells, e.g., reduction, delay, or inhibition of cell proliferation. A therapeutically effective amount can be determined by administering a low dose initially, then gradually increasing the dose until the desired effect is achieved. A therapeutically effective amount can vary depending on the intended use (in vitro or in vivo) or the subject and disease state being treated, e.g., the subject's weight and age, the severity of the disease state, the method of administration, etc., and can be readily determined by one of ordinary skill in the art. Specific amounts can vary depending, for example, on the particular pharmaceutical composition, the subject and their age and pre-existing health condition or risk for health conditions, the dosing regimen to be followed, the severity of the disease, whether it is administered in combination with other agents, the timing of administration, the tissue to which it is administered, and the physical delivery system by which it is delivered. In the case of cancer, a therapeutically effective amount of an ADC can reduce the number of cancer cells, reduce tumor size, inhibit (e.g., slow or stop) tumor metastasis, inhibit (e.g., slow or stop) tumor growth, and / or alleviate one or more symptoms.

[0343] As used herein, the term "prophylactically effective amount" or "prophylactically effective dose" refers to an amount of a compound disclosed herein, e.g., an ADC compound or composition described herein, effective at the dosage and for the period of time necessary to achieve the desired prophylactic result. Typically, a prophylactic dose is used in a subject prior to or at an early stage of disease, and therefore the prophylactically effective amount will be less than the therapeutically effective amount. In some embodiments, the prophylactically effective amount can prevent the onset of disease symptoms, including symptoms associated with cancer.

[0344] The term "p" or "drug loading" or "drug:antibody ratio" or "drug-to-antibody ratio" or "DAR" refers to the number of drug moieties per antibody or antigen-binding fragment, i.e., drug loading, or the number of -LD moieties per antibody or antigen-binding fragment (Ab) in an ADC of Formula (1). In an ADC containing an Mcl-1 inhibitor drug moiety, "p" refers to the number of Mcl-1 inhibitor compounds linked to the antibody or antigen-binding fragment. For example, if two Mcl-1 inhibitor compounds are linked to the antibody or antigen-binding fragment, p = 2. In a composition comprising multiple copies of an ADC of Formula (1), "average p" refers to the average number of -LD moieties per antibody or antigen-binding fragment, also referred to as "average drug loading."

[0345] Antibody-drug conjugates Antibody-drug conjugate (ADC) compounds of the present disclosure include those with anti-cancer activity. In particular, ADC compounds comprise an antibody or antigen-binding fragment conjugated (i.e., covalently attached via a linker) to a drug moiety (e.g., an Mcl-1 inhibitor), where the drug moiety has a cytotoxic or cytostatic effect when not conjugated to the antibody or antigen-binding fragment. In some embodiments, the drug moiety, when not conjugated to the antibody or antigen-binding fragment, is capable of reducing the expression and / or activity of Mcl-1 and / or one or more of its upstream regulators or downstream targets. Without being bound by theory, by targeting Mcl-1 expression and / or activity, in some embodiments, the ADCs disclosed herein can provide potent anti-cancer agents. Also, without being bound by theory, by conjugating the drug moiety to an antibody that binds to an antigen associated with tumor cell or cancer expression, the ADC can provide improved activity, better cytotoxic specificity, and / or reduced off-target killing compared to the drug moiety when administered alone.

[0346] In some embodiments, the components of the ADC are therefore selected to (i) retain one or more therapeutic properties exhibited by the isolated antibody and drug moiety, (ii) maintain the specific binding properties of the antibody or antigen-binding fragment; (iii) optimize drug loading and drug-to-antibody ratio; (iv) enable delivery of the drug moiety, e.g., intracellular delivery, via stable attachment to the antibody or antigen-binding fragment; (v) maintain stability of the ADC as an intact conjugate until translocation or delivery to the target site; (vi) minimize aggregation of the ADC before and after administration; (vii) enable the therapeutic effect, e.g., cytotoxic effect, of the drug moiety after cleavage or other release mechanism in the cellular environment; (viii) demonstrate in vivo anti-cancer treatment efficacy comparable to or superior to that of the isolated antibody and drug moiety; (ix) minimize off-target killing by the drug moiety; and / or (x) exhibit desirable pharmacokinetic and pharmacodynamic properties, formulatability, and toxicological / immunological profile. Each of these properties can provide improved ADCs for therapeutic use (Ab et al. (2015) Mol Cancer Ther. 14:1605-13).

[0347] The disclosed ADC compounds can selectively deliver an effective amount of a cytotoxic or cytostatic agent to cancer cells or tumor tissues. In some embodiments, the cytotoxic and / or cytostatic activity of the ADC depends on the expression of the target antigen in the cells. In some embodiments, the disclosed ADCs are particularly effective at killing cancer cells that express the target antigen while minimizing off-target killing. In some embodiments, the disclosed ADCs do not exhibit cytotoxic and / or cytostatic effects against cancer cells that do not express the target antigen.

[0348] In certain aspects, provided herein are ADC compounds comprising an antibody or antigen-binding fragment thereof (Ab) that targets a cancer cell, an Mcl-1 inhibitor drug moiety (D), and a linker moiety (L) covalently linking the Ab to D. In some embodiments, the antibody or antigen-binding fragment is capable of binding to a tumor-associated antigen (e.g., BCMA, CD33, PCAD, or HER2), e.g., with high specificity and high affinity. In some embodiments, the antibody or antigen-binding fragment is internalized into the target cell, e.g., into a degradable compartment within the cell, upon binding. In some embodiments, the ADC is internalized upon binding to the target cell and undergoes degradation, releasing the Mcl-1 inhibitor drug moiety and killing the cancer cell. The Mcl-1 inhibitor drug moiety can be released from the antibody and / or linker moiety of the ADC by enzymatic action, hydrolysis, oxidation, or any other mechanism.

[0349] A typical ADC is represented by the formula (1): Ab-(LD) p (1) where Ab = antibody or antigen-binding fragment, L = linker moiety, D = Mcl-1 inhibitor drug moiety, and p = number of Mcl-1 inhibitor drug moieties per antibody or antigen-binding fragment. It has.

[0350] antibody The antibody or antigen-binding fragment (Ab) of formula (1) includes within its scope any antibody or antigen-binding fragment that specifically binds to a target antigen on a cancer cell. The antibody or antigen-binding fragment may have a dissociation constant (K) of ≦1 mM, ≦100 nM, or ≦10 nM, or any amount therebetween, as measured, for example, by BIAcore® analysis. D ) and can bind to the target antigen. In some embodiments, K D In some embodiments, K D is between 500 pM and 1 μM, 1 μM and 100 nM, or 100 mM and 10 nM.

[0351] In some embodiments, the antibody or antigen-binding fragment is a four-chain antibody (also referred to as an immunoglobulin or a full-length or intact antibody), having two heavy chains and two light chains. In some embodiments, the antibody or antigen-binding fragment is an antigen-binding fragment of an immunoglobulin. In some embodiments, the antibody or antigen-binding fragment is an antigen-binding fragment of an immunoglobulin that retains the ability to bind to a target cancer antigen and / or provides at least one function of an immunoglobulin.

[0352] In some embodiments, the antibody or antigen-binding fragment is an internalizing antibody or internalizing antigen-binding fragment thereof. In some embodiments, the internalizing antibody or internalizing antigen-binding fragment thereof binds to a target cancer antigen expressed on the surface of a cell and enters the cell upon binding. In some embodiments, the Mcl-1 inhibitor drug moiety of the ADC is released from the antibody or antigen-binding fragment of the ADC after the ADC enters and resides in a cell expressing the target cancer antigen (i.e., after the ADC is internalized), e.g., by cleavage, degradation of the antibody or antigen-binding fragment, or any other suitable release mechanism.

[0353] [Table 3-1]

[0354] [Table 3-2]

[0355] [Table 3-3]

[0356] [Table 3-3]

[0357] [Table 3-4]

[0358] Table 4-1

[0359] Table 4-2

[0360] Table 4-3

[0361] Table 4-4

[0362] Table 4-5

[0363] Table 5-1

[0364] Table 5-2

[0365] Table 5-3

[0366] Table 5-4

[0367] Table 5-5

[0368] Table 5-6

[0369] Table 5-7

[0370] Table 5-8

[0371] Table 5-9

[0372] Table 5-10

[0373] Table 5-11

[0374] Table 5-12

[0375] Table 5-13

[0376] Table 5-14

[0377] Table 5-15

[0378] [Table 5-16]

[0379] [Table 5-17]

[0380] [Table 5-18]

[0381] [Table 5-19]

[0382] In some embodiments, the antibodies or antigen-binding fragments of the ADCs disclosed herein can comprise any set of heavy and light chain variable domains listed in the table above, or a set of six CDRs from any set of heavy and light chain variable domains listed in the table above. ... -8 K less than M D The ADCs may contain conservatively modified and / or homologous amino acid sequences to those listed in the table above, so long as they retain the ability to bind to the antigen (e.g., at 1000 .ANG.) and retain one or more functional properties of the ADCs disclosed herein (e.g., the ability to be internalized, to bind to an antigen target, e.g., an antigen expressed on a tumor or other cancer cell, etc.).

[0383] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 1, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 16, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 17, a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 19, a light chain CDR4 (LCDR4) consisting of SEQ ID NO: 20, a light chain CDR5 (LCDR5) consisting of SEQ ID NO: 21, a light chain CDR6 (LCDR6) consisting of SEQ ID NO: 22, a light chain CDR7 (LCDR7) consisting of SEQ ID NO: 23, a light chain C 70 and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18.

[0384] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 4, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 16, and a light chain CDR3 (LCDR1) consisting of SEQ ID NO: 17. 70 and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18.

[0385] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 4, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 16, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 17, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18.

[0386] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 19, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 20, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 21.

[0387] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 1, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 35, and a light chain CDR3 (LCDR1) consisting of SEQ ID NO: 36. 70and a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 17.

[0388] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 4, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 35, and a light chain CDR3 (LCDR1) consisting of SEQ ID NO: 36. 70 and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18.

[0389] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5; a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6; a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:3; 71 the light chain CDR1 (LCDR1) consisting of SEQ ID NO: 20, the light chain CDR2 (LCDR2) consisting of SEQ ID NO: 20, and the light chain CDR3 (LCDR3) consisting of SEQ ID NO: 21.

[0390] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 22, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 20, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 21.

[0391] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:8, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:17, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:20, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:18.

[0392] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 23. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 10, and the light chain variable region amino acid sequence of SEQ ID NO: 23, or a sequence at least 95% identical to a disclosed sequence. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 10, and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 23.

[0393] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 27. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 10 and the light chain variable region amino acid sequence of SEQ ID NO: 27, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 10 and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 27.

[0394] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 31. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 10 and the light chain variable region amino acid sequence of SEQ ID NO: 31, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 10 and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 31.

[0395] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 36. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 10 and the light chain variable region amino acid sequence of SEQ ID NO: 36, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 10 and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 36.

[0396] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 40. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 10 and the light chain variable region amino acid sequence of SEQ ID NO: 40, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 10 and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 40.

[0397] In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 44. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 10 and the light chain variable region amino acid sequence of SEQ ID NO: 44, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 10 and / or a light chain variable region amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 44.

[0398] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12, or a sequence at least 95% identical to SEQ ID NO: 12, and a light chain amino acid sequence of SEQ ID NO: 25, or a sequence at least 95% identical to SEQ ID NO: 25. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12 and a light chain amino acid sequence of SEQ ID NO: 25, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 12 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 25. In some embodiments, the anti-CD74 antibody is milatuzumab (see U.S. Patent No. 7,931,903) or an antigen-binding fragment thereof. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14, or a sequence at least 95% identical to SEQ ID NO: 14, and a light chain amino acid sequence of SEQ ID NO: 25, or a sequence at least 95% identical to SEQ ID NO: 25. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14 and a light chain amino acid sequence of SEQ ID NO: 25, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 14, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 25.

[0399] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15, or a sequence at least 95% identical to SEQ ID NO: 15, and a light chain amino acid sequence of SEQ ID NO: 25, or a sequence at least 95% identical to SEQ ID NO: 25. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15 and a light chain amino acid sequence of SEQ ID NO: 25, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 15, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 25.

[0400] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12, or a sequence at least 95% identical to SEQ ID NO: 12, and a light chain amino acid sequence of SEQ ID NO: 29, or a sequence at least 95% identical to SEQ ID NO: 29. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12 and a light chain amino acid sequence of SEQ ID NO: 29, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 12 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 29.

[0401] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14, or a sequence at least 95% identical to SEQ ID NO: 14, and a light chain amino acid sequence of SEQ ID NO: 29, or a sequence at least 95% identical to SEQ ID NO: 29. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14 and a light chain amino acid sequence of SEQ ID NO: 29, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 14, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 29.

[0402] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15, or a sequence at least 95% identical to SEQ ID NO: 15, and a light chain amino acid sequence of SEQ ID NO: 29, or a sequence at least 95% identical to SEQ ID NO: 29. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15 and a light chain amino acid sequence of SEQ ID NO: 29, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 15, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 29.

[0403] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12, or a sequence at least 95% identical to SEQ ID NO: 12, and a light chain amino acid sequence of SEQ ID NO: 33, or a sequence at least 95% identical to SEQ ID NO: 33. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12 and a light chain amino acid sequence of SEQ ID NO: 33, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 12 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 33.

[0404] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14, or a sequence at least 95% identical to SEQ ID NO: 14, and a light chain amino acid sequence of SEQ ID NO: 33, or a sequence at least 95% identical to SEQ ID NO: 33. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14 and a light chain amino acid sequence of SEQ ID NO: 33, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 14, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 33.

[0405] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15, or a sequence at least 95% identical to SEQ ID NO: 15, and a light chain amino acid sequence of SEQ ID NO: 33, or a sequence at least 95% identical to SEQ ID NO: 33. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15 and a light chain amino acid sequence of SEQ ID NO: 33, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 15 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 33.

[0406] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12, or a sequence at least 95% identical to SEQ ID NO: 12, and a light chain amino acid sequence of SEQ ID NO: 38, or a sequence at least 95% identical to SEQ ID NO: 38. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12 and a light chain amino acid sequence of SEQ ID NO: 38, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 12 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 38.

[0407] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14, or a sequence at least 95% identical to SEQ ID NO: 14, and a light chain amino acid sequence of SEQ ID NO: 38, or a sequence at least 95% identical to SEQ ID NO: 38. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14 and a light chain amino acid sequence of SEQ ID NO: 38, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 14, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 38.

[0408] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15, or a sequence at least 95% identical to SEQ ID NO: 15, and a light chain amino acid sequence of SEQ ID NO: 38, or a sequence at least 95% identical to SEQ ID NO: 38. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15 and a light chain amino acid sequence of SEQ ID NO: 38, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 15 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 38.

[0409] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12, or a sequence at least 95% identical to SEQ ID NO: 12, and a light chain amino acid sequence of SEQ ID NO: 42, or a sequence at least 95% identical to SEQ ID NO: 42. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12 and a light chain amino acid sequence of SEQ ID NO: 42, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 12 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 42.

[0410] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14, or a sequence at least 95% identical to SEQ ID NO: 14, and a light chain amino acid sequence of SEQ ID NO: 42, or a sequence at least 95% identical to SEQ ID NO: 42. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14 and a light chain amino acid sequence of SEQ ID NO: 42, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 14, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 42.

[0411] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15, or a sequence at least 95% identical to SEQ ID NO: 15, and a light chain amino acid sequence of SEQ ID NO: 42, or a sequence at least 95% identical to SEQ ID NO: 42. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15 and a light chain amino acid sequence of SEQ ID NO: 42, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 15, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 42.

[0412] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12, or a sequence at least 95% identical to SEQ ID NO: 12, and a light chain amino acid sequence of SEQ ID NO: 46, or a sequence at least 95% identical to SEQ ID NO: 46. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 12 and a light chain amino acid sequence of SEQ ID NO: 46, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 12 and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 46.

[0413] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14, or a sequence at least 95% identical to SEQ ID NO: 14, and a light chain amino acid sequence of SEQ ID NO: 46, or a sequence at least 95% identical to SEQ ID NO: 46. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 14 and a light chain amino acid sequence of SEQ ID NO: 46, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 14, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 46.

[0414] In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15, or a sequence at least 95% identical to SEQ ID NO: 15, and a light chain amino acid sequence of SEQ ID NO: 46, or a sequence at least 95% identical to SEQ ID NO: 46. In some embodiments, the anti-CD74 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 15 and a light chain amino acid sequence of SEQ ID NO: 46, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD74 antibody has a heavy chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 15, and a light chain amino acid sequence that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 46.

[0415] Residues in two or more polypeptides are said to "correspond" if they occupy similar positions in the polypeptide structure. Similar positions in two or more polypeptides can be determined by aligning the polypeptide sequences based on the similarity of amino acid sequence or structure. Those skilled in the art will understand that it may be necessary to introduce gaps into either sequence to produce a good alignment.

[0416] In some embodiments, amino acid substitutions are of a single residue. Insertions typically occur between about 1 and about 20 amino acid residues, although significantly larger insertions may be tolerated as long as biological function (e.g., binding to a target antigen) is maintained. Deletions typically range from about 1 to about 20 amino acid residues, although in some cases deletions may be much larger. Substitutions, deletions, insertions, or any combination thereof, can be used to arrive at a final derivative or variant. Generally, these changes are made on a small number of amino acids to minimize alterations in the immunogenicity and specificity of the molecule, particularly antigen-binding proteins. However, larger changes may be tolerated in certain circumstances. Conservative substitutions can be made according to the following chart, shown as Table 7:

[0417] [Table 6]

[0418] In some embodiments, in which variant antibody sequences are used in ADCs, the variants typically exhibit the same quality of biological activity and induce the same immune response, but the variants can also be selected to modify the properties of the antigen-binding protein as needed. Alternatively, the variants can be designed to alter the biological activity of the antigen-binding protein. For example, glycosylation sites can be altered or removed.

[0419] The immunoconjugates of the invention may comprise modified antibodies or antigen-binding fragments thereof further comprising modifications to framework residues within VH and / or VL, e.g., to improve the properties of the antibody. In some embodiments, framework modifications are made to reduce the immunogenicity of the antibody. For example, one approach is to "backmutate" one or more framework residues to the corresponding germline sequence. More specifically, antibodies that have undergone somatic mutation may contain framework residues that differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the antibody framework sequence with the germline sequence from which the antibody is derived. Somatic mutations can be "backmutated" to the germline sequence, e.g., by site-directed mutagenesis, to return the framework region sequences to their germline configuration. Such "backmutated" antibodies are also intended to be encompassed by the present invention.

[0420] Another type of framework modification involves mutating one or more residues within the framework regions, or even one or more CDR regions, to remove T-cell epitopes, thereby reducing the potential immunogenicity of the antibody. This approach, also known as "deimmunization," is described in further detail in U.S. Patent Publication No. 20030153043 by Carr et al.

[0421] In addition to, or instead of, modifications made in the framework or CDR regions, antibodies of the invention may be engineered to contain modifications in the Fc region, typically to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or antigen-dependent cellular cytotoxicity (ADCC). Further, antibodies of the invention may be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or modified to alter its glycosylation, again to alter one or more functional properties of the antibody. Each of these embodiments is described in further detail below.

[0422] In one embodiment, the hinge region of CH1 is modified so that the number of cysteine ​​residues in the hinge region is altered, e.g., increased or decreased. This approach is further described in U.S. Patent No. 5,677,425 by Bodmer et al. The number of cysteine ​​residues in the hinge region of CH1 is altered, for example, to facilitate assembly of the light and heavy chains or to improve or decrease the stability of the antibody.

[0423] In some embodiments, the antibodies or antibody fragments disclosed herein contain modified or engineered amino acid residues, such as one or more cysteine ​​residues, as sites for conjugation with a drug moiety (Junutula JR, et al., Nat Biotechnol 2008, 26:925-932). In one embodiment, the present invention provides modified antibodies or antibody fragments containing substitutions of one or more amino acids with cysteine ​​at the positions described herein. The sites for cysteine ​​substitutions are within the constant region of the antibody or antibody fragment and are therefore applicable to a variety of antibodies or antibody fragments, with the sites selected to provide stable and homogeneous conjugates. The modified antibodies or fragments can have one or two or more cysteine ​​substitutions, and these substitutions can be used in combination with other modification and conjugation methods as described herein. Methods for inserting cysteines at specific positions in antibodies are known in the art, see, e.g., Lyons et al., (1990) Protein Eng., 3:703-708, WO 2011 / 005481, WO 2014 / 124316, WO 2015 / 138615. In certain embodiments, the modified antibody comprises a substitution of one or more amino acids with cysteines in its constant region selected from positions 117, 119, 121, 124, 139, 152, 153, 155, 157, 164, 169, 171, 174, 189, 191, 195, 197, 205, 207, 246, 258, 269, 274, 286, 288, 290, 292, 293, 320, 322, 326, 333, 334, 335, 337, 344, 355, 360, 375, 382, ​​390, 392, 398, 400 and 422 of the heavy chain of the antibody, where positions are numbered according to the EU system.In some embodiments, the modified antibody or antibody fragment comprises a substitution of one or more amino acids with cysteine ​​on its constant region selected from positions 107, 108, 109, 114, 129, 142, 143, 145, 152, 154, 156, 159, 161, 165, 168, 169, 170, 182, 183, 197, 199, and 203 of the light chain of the antibody or antibody fragment, where the positions are numbered according to the EU system, and where the light chain is a human kappa light chain. In certain embodiments, the modified antibody or antibody fragment thereof comprises a combination of substitutions of two or more amino acids with cysteine ​​on its constant region, where the combination includes a substitution at position 375 of the antibody heavy chain, position 152 of the antibody heavy chain, position 360 of the antibody heavy chain, or position 107 of the antibody light chain, where positions are numbered according to the EU system. In certain embodiments, the modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine ​​on its constant region, wherein the substitution is at position 375 of the antibody heavy chain, position 152 of the antibody heavy chain, position 360 of the antibody heavy chain, position 107 of the antibody light chain, position 165 of the antibody light chain, or position 159 of the antibody light chain, where positions are numbered according to the EU system, and where the light chain is a kappa chain. In certain embodiments, the modified antibody or antibody fragment thereof comprises a combination of substitutions of two amino acids with cysteine ​​on its constant region, wherein the combination comprises substitutions at position 375 of the antibody heavy chain and position 152 of the antibody heavy chain, where positions are numbered according to the EU system. In certain embodiments, the modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine ​​at position 360 of the antibody heavy chain, where positions are numbered according to the EU system. In other specific embodiments, the modified antibody or antibody fragment thereof comprises a substitution of one amino acid with a cysteine ​​at position 107 of the antibody light chain, where positions are numbered according to the EU system, and where the light chain is a kappa chain.

[0424] In additional embodiments, antibodies or antibody fragments (e.g., antigen-binding fragments) useful in the immunoconjugates of the invention include modified or engineered antibodies, such as antibodies modified to introduce one or more other reactive amino acids (other than cysteine), including PcI, pyrrolysine, peptide tags (such as S6, A1 and ybbR tags), and a non-natural amino acid in place of at least one amino acid of the native sequence, thus providing a reactive site on the antibody or antigen-binding fragment for conjugation with a drug moiety or linker-drug moiety of complementary reactivity. For example, antibodies or antibody fragments can be modified to incorporate Pcl or pyrrolysine (W. Ou, et al., (2011) PNAS 108 (26), 10437-10442; WO 2014124258) or unnatural amino acids (JY Axup, et al., Proc Natl Acad Sci USA, 109 (2012), pp. 16101-16106; for reviews, see CC Liu and PG Schultz (2010) Annu Rev Biochem 79, 413-444; CH Kim, et al., (2013) Curr Opin Chem Biol. 17, 412-419) as sites for conjugation with drugs. Similarly, peptide tags for enzymatic conjugation can be introduced into antibodies (Strop P., et al., Chem Biol. 2013, 20(2):161-7; Rabuka D., Curr Opin Chem Biol. 2010 Dec;14(6):790-6; Rabuka D., et al., Nat Protoc. 2012, 7(6):1052-67). Another example is the use of 4'-phosphopantetheinyl transferase (PPTase) for the conjugation of coenzyme A analogs (WO2013184514) and (Grunewald et al., (2015) Bioconjugate Chem. 26(12), 2554-62).Methods for conjugating such modified or engineered antibodies with a payload or linker-payload combination are known in the art.

[0425] In another embodiment, the Fc-hinge region of an antibody is mutated to shorten the biological half-life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment so that the antibody has impaired Staphylococcus protein A (SpA) binding relative to native Fc-hinge domain SpA binding. This approach is described in further detail in U.S. Patent No. 6,165,745 by Ward et al.

[0426] In yet other embodiments, the Fc region is altered by replacing at least one amino acid residue with a different amino acid residue to alter the effector function of the antibody. For example, one or more amino acids can be replaced with different amino acid residues so that the antibody has altered affinity for an effector ligand but retains the antigen-binding ability of the parent antibody. The effector ligand to which affinity is altered can be, for example, an Fc receptor or the C1 component of complement. This approach is described, for example, in U.S. Patent Nos. 5,624,821 and 5,648,260, both by Winter et al.

[0427] In another embodiment, one or more selected amino acids can be replaced with different amino acid residues such that the antibody has altered C1q binding and / or reduced or abolished complement dependent cytotoxicity (CDC), as described, for example, in U.S. Patent No. 6,194,551 to Idusogie et al.

[0428] In another embodiment, one or more amino acid residues are altered to thereby alter the antibody's ability to fix complement. This approach is described, for example, in PCT Publication WO 94 / 29351 by Bodmer et al. Allotypic amino acid residues include, but are not limited to, the heavy chain constant regions of the IgG1, IgG2, and IgG3 subclasses and the light chain constant region of the kappa isotype, as described by Jefferis et al., MAbs. 1:332-338 (2009).

[0429] In some embodiments, the antibody contains mutations that reduce or completely mediate antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC). In some embodiments, these mutations are known as Fc silencing, Fc silent, or Fc silent mutations. In some embodiments, amino acid residues L234 and L235 in the IgG1 constant region are substituted with A234 and A235 (also known as "LALA"). In some embodiments, amino acid residue N297 in the IgG1 constant region is substituted with A297 (also known as "N297A"). In some embodiments, amino acid residues D265 and P329 in the IgG1 constant region are substituted with A265 and A329 (also known as "DAPA"). Other antibody Fc silencing mutations can also be used. In some embodiments, Fc silencing mutations are used in combination, for example, D265A, N297A, and P329A (also known as "DANAPA").

[0430] In another embodiment, one or more amino acid residues are altered to thereby alter the antibody's ability to fix complement. This approach is described, for example, in PCT Publication WO 94 / 29351 by Bodmer et al. In certain embodiments, one or more amino acids of an antibody or antigen-binding fragment thereof of the present invention are replaced with one or more allotypic amino acid residues. Allotypic amino acid residues include, but are not limited to, the heavy chain constant regions of IgG1, IgG2, and IgG3 subclasses and the light chain constant region of the kappa isotype, as described by Jefferis et al., MAbs. 1:332-338 (2009).

[0431] In yet another embodiment, the glycosylation of the antibody is modified. For example, an aglycosylated antibody can be generated (i.e., the antibody lacks glycosylation). Glycosylation can be altered to, for example, increase the affinity of the antibody for an "antigen." Such carbohydrate modifications can be achieved, for example, by altering one or more sites of glycosylation within the antibody sequence. For example, one or more amino acid substitutions can be made that result in the elimination of one or more variable region framework glycosylation sites, thereby eliminating glycosylation at that site. Such aglycosylation can increase the affinity of the antibody for the antigen. Such techniques are described, for example, in U.S. Patent Nos. 5,714,350 and 6,350,861 by Co et al.

[0432] In another embodiment, the antibody is modified to increase its biological half-life. Various approaches are possible. For example, as described in U.S. Patent No. 6,277,375 by Ward, one or more of the following mutations can be introduced: T252L, T254S, T256F. Alternatively, to increase biological half-life, the antibody can be modified in the CH1 or CL region to include salvage receptor binding epitopes taken from two loops of the CH2 domain of the Fc region of IgG, as described in U.S. Patent Nos. 5,869,046 and 6,121,022 by Presta et al.

[0433] Linker In some embodiments, the linker in the ADC is sufficiently extracellularly stable to be therapeutically effective. In some embodiments, the linker is extracellularly stable such that the ADC remains intact when present in extracellular conditions (e.g., before transport or delivery to a cell). The term "intact" as used in the context of an ADC means that the antibody or antigen-binding fragment remains attached to the drug moiety (e.g., an Mcl-1 inhibitor).

[0434] As used herein, "stable" in the context of a linker, or an ADC comprising a linker, means that when the ADC is present in extracellular conditions, no more than about 20%, no more than about 15%, no more than about 10%, no more than about 5%, no more than about 3%, or no more than about 1% (or any percentage therebetween) of the linkers in a sample of the ADC are cleaved (or the entire ADC is not otherwise intact). In some embodiments, the linkers and / or ADCs disclosed herein are more stable than ADCs with alternative linkers and / or alternative linkers and / or Mcl-1 inhibitor payloads. In some embodiments, the ADCs disclosed herein may remain intact for more than about 48 hours, more than 60 hours, more than about 72 hours, more than about 84 hours, or more than about 96 hours.

[0435] Whether a linker is stable extracellularly can be determined, for example, by placing the ADC in plasma for a predetermined period of time (e.g., 2, 4, 6, 8, 16, 24, 48, or 72 hours) and then quantifying the amount of free drug moiety present in the plasma. Stability may allow the ADC time to localize to target cancer cells and prevent premature release of the drug moiety, which could reduce the therapeutic index of the ADC by indiscriminately damaging both normal and cancer tissues. In some embodiments, the linker is stable outside the target cell and releases the drug moiety from the ADC once inside the cell, allowing the drug to bind to its target. Thus, an effective linker: (i) maintains the specific binding properties of the antibody or antigen-binding fragment; (ii) enables delivery of the drug moiety, e.g., intracellular delivery, via stable binding to the antibody or antigen-binding fragment; (iii) remains stable and intact until the ADC is transported or delivered to its target site; and (iv) enables the therapeutic effect, e.g., cytotoxic effect, of the drug moiety after cleavage or an alternative release mechanism.

[0436] Linkers can affect the physicochemical properties of ADCs. Because many cytotoxic agents are hydrophobic in nature, linking them to antibodies with additional hydrophobic moieties can result in aggregation. ADC aggregates are often insoluble and limit the achievable drug loading on the antibody, which can negatively impact the efficacy of the ADC. Protein aggregation of biologics is also commonly associated with increased immunogenicity. As shown below, the linkers disclosed herein result in ADCs with low aggregation levels and desirable drug loading levels.

[0437] Linkers can be "cleavable" or "non-cleavable" (Ducry and Stump (2010) Bioconjugate Chem. 21:5-13). Cleavable linkers are designed to release the drug moiety (e.g., an Mcl-1 inhibitor) when subjected to certain environmental factors, such as when internalized within target cells, whereas non-cleavable linkers generally rely on degradation of the antibody or antigen-binding fragment itself.

[0438] The term "alkyl," as used herein, refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms and containing no unsaturation. The term "C1-C6 alkyl," as used herein, refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms and containing no unsaturation, having from 1 to 6 carbon atoms, and attached to the remainder of the molecule by a single bond. Non-limiting examples of "C1-C6 alkyl" groups include methyl (C1 alkyl), ethyl (C2 alkyl), 1-methylethyl (C3 alkyl), n-propyl (C3 alkyl), isopropyl (C3 alkyl), n-butyl (C4 alkyl), isobutyl (C4 alkyl), sec-butyl (C4 alkyl), tert-butyl (C4 alkyl), n-pentyl (C5 alkyl), isopentyl (C5 alkyl), neopentyl (C5 alkyl), and hexyl (C6 alkyl).

[0439] The term "alkenyl," as used herein, refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms and containing at least one double bond. The term "C2-C6 alkenyl," as used herein, refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from 2 to 6 carbon atoms and attached to the rest of the molecule by a single bond. Non-limiting examples of "C2-C6 alkenyl" groups include ethenyl (C2 alkenyl), prop-1-enyl (C3 alkenyl), but-1-enyl (C4 alkenyl), pent-1-enyl (C5 alkenyl), pent-4-enyl (C5 alkenyl), penta-1,4-dienyl (C5 alkenyl), hex-1-enyl (C6 alkenyl), hex-2-enyl (C6 alkenyl), hex-3-enyl (C6 alkenyl), hexa-1,4-dienyl (C6 alkenyl), hexa-1,5-dienyl (C6 alkenyl), and hexa-2,4-dienyl (C6 alkenyl). The term "C2-C3 alkenyl," as used herein, refers to a straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, having 2 to 3 carbon atoms, and attached to the rest of the molecule by a single bond. Non-limiting examples of "C2-C3 alkenyl" groups include ethenyl (C2 alkenyl) and prop-1-enyl (C3 alkenyl).

[0440] The term "alkylene," as used herein, refers to a divalent straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms and containing no unsaturation. The term "C1-C6 alkylene," as used herein, refers to a divalent straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, containing no unsaturation, and having from 1 to 6 carbon atoms. Non-limiting examples of "C1-C6 alkylene" groups include methylene (C1 alkylene), ethylene (C2 alkylene), 1-methylethylene (C3 alkylene), n-propylene (C3 alkylene), isopropylene (C3 alkylene), n-butylene (C4 alkylene), isobutylene (C4 alkylene), sec-butylene (C4 alkylene), tert-butylene (C4 alkylene), n-pentylene (C5 alkylene), isopentylene (C5 alkylene), neopentylene (C5 alkylene), and hexylene (C6 alkylene).

[0441] The term "alkenylene," as used herein, refers to a divalent straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms and containing at least one double bond. The term "C2-C6 alkenylene," as used herein, refers to a divalent straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one double bond, and having from 2 to 6 carbon atoms. Non-limiting examples of "C2-C6 alkenylene" groups include ethenylene (C2 alkenylene), prop-1-enylene (C3 alkenylene), but-1-enylene (C4 alkenylene), pent-1-enylene (C5 alkenylene), pent-4-enylene (C5 alkenylene), penta-1,4-dienylene (C5 alkenylene), hex-1-enylene (C6 alkenylene), hex-2-enylene (C6 alkenylene), hex-3-enylene (C6 alkenylene), hexa-1-,4-dienylene (C6 alkenylene), hexa-1-,5-dienylene (C6 alkenylene), and hexa-2-,4-dienylene (C6 alkenylene). The term "C2-C6 alkenylene," as used herein, refers to a divalent straight or branched hydrocarbon chain radical group consisting solely of carbon and hydrogen atoms, containing at least one double bond, and having 2 to 3 carbon atoms. Non-limiting examples of "C2-C3 alkenylene" groups include ethenylene (C2 alkenylene) and prop-1-enylene (C3 alkenylene).

[0442] The term "cycloalkyl" or "C3-C8 cycloalkyl," as used herein, refers to a saturated, monocyclic, fused bicyclic, fused tricyclic, or bridged polycyclic ring system. Non-limiting examples of fused bicyclic or bridged polycyclic ring systems include bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[3.1.1]heptane, bicyclo[3.2.1]octane, bicyclo[2.2.2]octane, and adamantanyl. Non-limiting examples of monocyclic C3-C8 cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups.

[0443] The term "haloalkyl" as used herein refers to a linear or branched alkyl chain in which one or more halogen groups are substituted for hydrogen along the hydrocarbon chain. Examples of halogen groups suitable for substitution in haloalkyl groups include fluorine, bromine, chlorine, and iodine. A haloalkyl group may contain multiple halogen groups substituted for hydrogen in the alkyl chain, where the halogen groups can be bonded to the same carbon or different carbons in the alkyl chain.

[0444] As used herein, alkyl, alkenyl, alkynyl, alkoxy, amino, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl groups include optionally substituted linear or branched (C1-C6) alkyl, optionally substituted linear or branched (C2-C6) alkenyl groups, optionally substituted linear or branched (C2-C6) alkynyl groups, optionally substituted linear or branched (C1-C6) alkoxy, optionally substituted (C1-C6) alkyl-S-, hydroxy, oxo (or N-oxide, as appropriate), nitrite, ... and optionally substituted with 1 to 4 groups selected from phenyl, cyano, —C(O)—OR′, —OC(O)—R′, —C(O)—NR′R′, —NR′R′, —(C═NR′)—OR′, linear or branched (C1-C6) haloalkyl, trifluoromethoxy, or halogen, wherein R′ and R′ are each independently a hydrogen atom or an optionally substituted linear or branched (C1-C6) alkyl group, and wherein one or more of the carbon atoms of the linear or branched (C1-C6) alkyl group are optionally deuterated.

[0445] The terms "polyoxyethylene," "polyethylene glycol," or "PEG," as used herein, refer to a linear, branched, or star-shaped construct composed of (OCHCH) groups. In certain embodiments, the polyethylene or PEG group is -(OCHCH) t * - (wherein t is 4 to 40, where "-" indicates the end directed toward the self-immolative spacer, and * The "-" indicates the point of attachment to the terminal group R', where R' is OH, OCH, or OCHCHC(=O)OH. In other embodiments, the polyethylene or PEG group is -(CHCHO). t * - (wherein t is 4 to 40, where "-" indicates the end directed toward the self-immolative spacer, and *The "-" indicates the point of attachment to the terminal group R", where R" is H, CH, or CHCHC(=O)OH. For example, the term "PEG12", as used herein, means that t is 12.

[0446] The term "polyalkylene glycol" as used herein means (O(CH) m ) n In certain embodiments, a polyethylene or PEG group refers to a linear, branched, or star-shaped configuration composed of —O(CH) groups. m ) t * - (wherein m is 1 to 10 and t is 4 to 40, where "-" indicates the end directed toward the self-immolative spacer, * The "-" indicates the point of attachment to the terminal group R', where R' is OH, OCH, or OCHCHC(=O)OH. In other embodiments, the polyethylene or PEG group is -((CH) m O) t * - (wherein m is 1 to 10 and t is 4 to 40, where "-" indicates the end directed toward the self-immolative spacer, * The "-" indicates the point of attachment to the terminal group R", where R" is H, CH3, or CH2CH2C(=O)OH.

[0447] The term "reactive group," as used herein, refers to a functional group that can form a covalent bond with a functional group of an antibody, antibody fragment, or another reactive group attached to an antibody or antibody fragment. Non-limiting examples of such functional groups include the reactive groups in Table 2 provided herein.

[0448] The term "attachment group" or "coupling group," as used herein, refers to a bivalent moiety that links a bridging spacer and an antibody or fragment thereof. The bond or linking group is a bivalent moiety formed by the reaction between a reactive group and a functional group on an antibody or fragment thereof. Non-limiting examples of such bivalent moieties include the bivalent chemical moieties shown in Tables 2 and 3 provided herein.

[0449] The term "bridge spacer," as used herein, refers to one or more linker components that are covalently linked together to form a bivalent moiety that connects a bivalent peptide spacer to a reactive group, a bivalent peptide spacer to a linking group, or a linking group to at least one cleavable group. In certain embodiments, a "bridge spacer" comprises a carboxyl group that is attached to the N-terminus of the bivalent peptide spacer via an amide bond.

[0450] The term "spacer moiety," as used herein, refers to one or more linker components that are covalently bonded together to form the moiety that connects the self-immolative spacer and the hydrophilic moiety.

[0451] The term "bivalent peptide spacer," as used herein, refers to a bivalent linker that includes one or more amino acid residues that are covalently linked together to form the moiety that connects the bridging spacer and the self-immolative spacer. The one or more amino acid residues may be residues of an amino acid selected from alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), norvaline (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine, and desmethylpyrrolysine.

[0452] In certain embodiments, a "bivalent peptide spacer" is a peptide spacer in which each residue is selected from the group consisting of alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (A a combination of 2 to 4 amino acid residues independently selected from amino acid residues selected from amino acid residues selected from: -Val, -Ser, -Thr, -Val, -Valg, -Trp, -Tyr, -Cit, -Nva, -Norleucine (Nle), -Selenocysteine ​​(Sec), -Pyrrolysine (Pyl), -Homoserine, -Homocysteine, and -Desmethylpyrrolysine, for example, -ValCit * ;-CitVal * ;-AlaAla * ;-AlaCit * ;-CitAla * ;-AsnCit *;-CitAsn * ;-CitCit * ;-ValGlu * ;-GluVal * ;-SerCit * ;-CitSer * ;-LysCit * ;-CitLys * ;-AspCit * ;-CitAsp * ;-AlaVal * ;-ValAla * ;-PheAla * ;-AlaPhe * ;-PheLys * ;-LysPhe * ;-ValLys * ;-LysVal * ;-AlaLys * ;-LysAla * ;-PheCit * ;-CitPhe * ;-LeuCit * ;-CitLeu * ;-IleCit * ;-CitIle * ;-PheArg * ;-ArgPhe * ;-CitTrp * ;-TrpCit * ;-PhePheLys * ;-LysPhePhe * ;-DPhePheLys * ;-DLysPhePhe * ;-GlyPheLys * ;-LysPheGly * -GlyPheLeuGly-[SEQ ID NO: 69]; -GlyLeuPheGly-[SEQ ID NO: 64]; -AlaLeuAlaLeu-[SEQ ID NO: 65], -GlyGlyGly * -GlyGlyGlyGly-[SEQ ID NO: 66]; -GlyPheValGly-[SEQ ID NO: 67]; and -GlyValPheGly-[SEQ ID NO: 68], where "-" indicates the point of attachment to the bridging spacer. *" indicates the point of attachment to the self-immolative spacer.

[0453] The term "linker moiety," as used herein, refers to a chemical moiety that is part of a linker. Examples of linker moieties include: alkylene groups: -(CH), which can be either linear or branched. n - (where n is 1 to 18); alkenylene group; alkynylene group; alkenyl group; alkynyl group; ethylene glycol unit: -OCH2CH2- or -CH2CH2O-; polyethylene glycol unit: (-CH2CH2O-) x (wherein x is 2 to 20); -O-; -S-; carbonyl: -C(=O); ester: C(=O)-O or OC(=O); carbonate: -OC(=O)O-; amine: -NH-; tertiary amine; amide: -C(=O)-NH-, -NH-C(=O)- or -C(=O)N(C 1~6 alkyl); carbamate: -OC(=O)NH- or -NHC(=O)O; urea: -NHC(=O)NH; sulfonamide: -S(O)NH- or -NHS(O)2; ether: -CHO- or -OCH2-; alkylene substituted with one or more groups independently selected from carboxy, sulfonic acid, hydroxyl, amine, amino acid, sugar, phosphoric acid, and phosphonic acid; alkenylene substituted with one or more groups independently selected from carboxy, sulfonic acid, hydroxyl, amine, amino acid, sugar, phosphoric acid, and phosphonic acid; alkynylene substituted with one or more groups independently selected from carboxy, sulfonic acid, hydroxyl, amine, amino acid, sugar, phosphoric acid, and phosphonic acid; C1-C in which one or more methylene groups are replaced with one or more -S-, -NH-, or -O- moieties. 10alkylene; a bivalent ring selected from ring systems having two available points of attachment, such as phenyl (including 1,2-, 1,3-, and 1,4-disubstituted phenyl), C5-C6 heteroaryl, C3-C8 cycloalkyl (including 1,1-disubstituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, and 1,4-disubstituted cyclohexyl), and C4-C8 heterocycloalkyl; alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Ly), residues of amino acids selected from the group consisting of alanine (Ala), cysteine ​​(Cla), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), norvaline (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine, and desmethylpyrrolysine; Cys, aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), norvaline (Nva), norleucine (Nle ... Combinations of two or more amino acid residues independently selected from amino acid residues selected from Sec, pyrrolysine (Pyl), homoserine, homocysteine, and desmethylpyrrolysine, e.g., Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala;Phe-Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu-Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit; and self-immolative spacers comprising one or more protecting (trigger) groups that are susceptible to acid-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, glycosidase-induced cleavage, phosphodiesterase-induced cleavage, phosphatase-induced cleavage, protease-induced cleavage, lipase-induced cleavage, or disulfide bond cleavage.

[0454] Non-limiting examples of such self-immolative spacers include:

[0455] [ka] (In the formula, PG is a protecting (trigger) group, X a is O, NH or S, X b is O, NH, NCH3 or S, X c is O or NH, Y a is CH2, CH2O or CH2NH, Y b is CH2, O or NH, Y c is a bond, CH, O, or NH, LG is a leaving group, such as the drug moiety (D) of a linker-drug group of the invention. Examples include: Additional non-limiting examples of such self-immolative spacers are described in Angew. Chem. Int. Ed. 2015, 54, 7492-7509.

[0456] Additionally, a linker component can be a chemical moiety that is readily formed by a reaction between two reactive groups. Non-limiting examples of such chemical moieties are shown in Table 2.

[0457] [Table 7-1]

[0458] [Table 7-2]

[0459] [Table 7-3]

[0460] [Table 7-4]

[0461] [Table 7-5] Here, R in Table 2 32 is H, C 1~4 alkyl, phenyl, pyrimidine or pyridine, and R in Table 2 35 is H, C 1~6 C substituted with alkyl, phenyl, or 1 to 3 -OH groups 1~4 alkyl, and each R in Table 2 7 is H, C 1~6 Alkyl, fluoro, benzyloxy substituted with -C(=O)OH, benzyl substituted with -C(=O)OH, C substituted with -C(=O)OH 1~4 C substituted with alkoxy and -C(=O)OH 1~4 alkyl; 37 are independently selected from H, phenyl and pyridine, q in Table 2 is 0, 1, 2 or 3, and R in Table 2 8 and R 13 are H or methyl, and R in Table 29 and R 14 are each H, —CH or phenyl, R in Table 2 is H or any suitable substituent, and R in Table 2 50 is H.

[0462] In addition, the linker moiety can be a group listed in Table 3 below.

[0463] [Table 8-1]

[0464] [Table 8-2]

[0465] [Table 8-3]

[0466] As used herein, when a substructure of a compound is illustrated, it is indicated by a wavy line (

[0467] [ka] ) indicates the point of attachment of the substructure to the rest of the molecule.

[0468] The terms "self-immolative spacer" and "self-immolative group," as used herein, refer to a moiety containing one or more trigger groups (TG) that are activated by acid-, peptidase-, esterase-, glycosidase-, phosphodiesterase-, phosphatase-, protease-, lipase-, or disulfide bond cleavage, whereupon the protecting groups are removed, resulting in a cascade of decomposition reactions that lead to the time-sequential release of leaving groups. Such a cascade of reactions can be, but is not limited to, 1,4-, 1,6-, or 1,8-elimination reactions.

[0469] Non-limiting examples of self-immolative spacers or groups include:

[0470] [ka] (wherein such groups may be optionally substituted, During the ceremony, TG is a trigger group, X a is O, NH or S, X b is O, NH, NCH3 or S, X c is O or NH, Y a is CH2, CH2O or CH2NH, Y b is CH2, O or NH, Y c is a bond, CH, O, or NH, LG is a leaving group, such as the drug moiety (D) of a linker-drug group of the invention. Examples include: Additional non-limiting examples of self-immolative spacers are described in Angew. Chem. Int. Ed. 2015, 54, 7492-7509.

[0471] In certain embodiments, the self-immolative spacer has the structure

[0472] [ka] where Lp is an enzyme-cleavable bivalent peptide spacer, and A, D, L3 and R 2 are as defined herein) It is a part having the following.

[0473] In a preferred embodiment, the self-immolative spacer has the structure

[0474] [ka] where Lp is an enzyme-cleavable bivalent peptide spacer, and D, L3 and R 2 are as defined herein) In some embodiments, D is a quaternized tertiary amine containing MCl1 inhibitor.

[0475] In another preferred embodiment, the self-immolative spacer has the structure

[0476] [ka] where Lp is an enzyme-cleavable bivalent peptide spacer, and D, L3 and R 2 are as defined herein) It is a part having the following.

[0477] The term "hydrophilic moiety," as used herein, refers to a moiety that has hydrophilic properties that increase the water solubility of the drug moiety (D) when the drug moiety (D) is attached to a linker group of the present invention. Examples of such hydrophilic groups include, but are not limited to, polyethylene glycol, polyalkylene glycol, saccharides, oligosaccharides, polypeptides, 1-3 alkyl groups, and the like.

[0478] [ka] Examples include C2 to C6 alkyl substituted with a group.

[0479] Drug portion In some embodiments, an intermediate that is a precursor to the linker moiety is reacted with a drug moiety (e.g., an Mcl-1 inhibitor) under appropriate conditions. In some embodiments, a reactive group is used on the drug and / or intermediate or linker. The product of the reaction between the drug and intermediate, or the derivatized drug (drug plus linker), is then reacted with an antibody or antigen-binding fragment under conditions that facilitate conjugation of the drug and intermediate or derivatized drug to the antibody or antigen-binding fragment. Alternatively, the intermediate or linker may be first reacted with an antibody or antigen-binding fragment, or a derivatized antibody or antigen-binding fragment, and then reacted with the drug or derivatized drug.

[0480] Several different reactions are available for covalently linking drug moieties and / or linker moieties to antibodies or antigen-binding fragments. This is often achieved by reacting one or more amino acid residues of antibodies or antigen-binding fragments, including the amine groups of lysine, the free carboxylic acid groups of glutamic acid and aspartic acid, the sulfhydryl groups of cysteine, and various aromatic amino acid residues. For example, nonspecific covalent linkages can be performed using a carbodiimide reaction to link a carboxy (or amino) group on the drug moiety with an amino (or carboxy) group on the antibody or antigen-binding fragment. In addition, bifunctional agents such as dialdehydes or imidoesters may also be used to link amino groups on the drug moiety with amino groups on the antibody or antigen-binding fragment. The Schiff base reaction can also be used to link drugs (e.g., Mcl-1 inhibitors) with binders. This method involves periodate oxidation of drugs containing glycol or hydroxy groups to form aldehydes, which are then reacted with binders. The binding occurs via the formation of a Schiff base with the amino group of the binding agent. Isothiocyanates can also be used as coupling agents to covalently bind drugs to binding agents. Other techniques are known to those skilled in the art and are within the scope of the present disclosure. Examples of drug moieties that can be generated and linked to antibodies or antigen-binding fragments using various chemistries known in the art include Mcl-1 inhibitors, such as the Mcl-1 inhibitors described and exemplified herein.

[0481] Suitable drug moieties may include compounds of Formula (I), (II), (III), or their enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable acid or base addition salts. In addition, the drug moiety may include any of the Mcl-1 inhibitors (D) described herein.

[0482] As used herein, an "atropisomer" is a stereoisomer that arises due to restricted rotation about a single bond when the energy difference due to steric strain or other contributors creates a barrier to rotation high enough to allow isolation of individual conformers (Bringmann et al. Angew. Chem. Int. Ed. 2005, 44, 5384-5427). For example, for a compound of formula (II) according to the present invention, the atropisomers may be:

[0483] [ka]

[0484] For example, the preferred atropisomer is (5aS), also referred to as (5S a ) can be.

[0485] The drug moieties of the present disclosure may be any of the compounds disclosed in International Patent Application Publication Nos. WO2015 / 097123; WO2016 / 207216; WO2016 / 207217; WO2016 / 207225; WO2016 / 207226; WO2017 / 125224; WO2019 / 035899; WO2019 / 035911; WO2019 / 035914; WO and WO2017 / 182625, and U.S. Patent Application Publication No. 2019 / 0055264 (each of which is incorporated herein by reference in its entirety).

[0486] In some embodiments, the drug moiety of the present disclosure has formula (I):

[0487] [ka] (In the formula, Ring D0 is a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group; Ring E0 is a furyl, thienyl or pyrrolyl ring; X 01 , X 03 , X 04 and X 05 are each independently a carbon atom or a nitrogen atom, X 02 is CR 026 group or a nitrogen atom,

[0488] [ka] means that the ring is aromatic, Y0 is a nitrogen atom or CR 03 It is the basis, Z0 is a nitrogen atom or CR 04 It is the basis, R 01is a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl group, a hydroxy group, a hydroxy(C1-C6) alkyl group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, -Cy 08 , -(C0-C6) alkyl-NR 011 R 011 ', -O-(C1-C6) alkyl-NR 011 R 011 ', -O-(C1-C6) alkyl-R 012 , -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ' or -SO2-(C1-C6)alkyl; R 02 , R 03 , R 04 and R 05 are each independently a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl, a hydroxy group, a hydroxy(C1-C6) alkyl group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, or a -(C0-C6) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C0-C6) alkyl-Cy 01 , -(C2-C6)alkenyl-Cy 01 , -(C2-C6)alkynyl-Cy 01, -O-(C1-C6) alkyl-NR 011 R 011 ', -O-(C1-C6) alkyl-R 031 , -O-(C1-C6)alkyl-R 012 , -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ',or -SO2-(C1-C6) alkyl, Or pair (R 01 , R 02 ), (R 02 , R 03 ), (R 03 , R 04 ) or (R 04 , R 05 ) together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring may be substituted with halogen, linear or branched (C1-C6) alkyl, (C0-C6) alkyl-NR 011 R 011 ', -NR 013 R 013 ', -(C0-C6) alkyl-Cy 01 or oxo, R 06 and R 07are each independently a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl, a hydroxy group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, or a -(C0-C6) alkyl-NR 011 R 011 ', -O-(C1-C6) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C0-C6) alkyl-Cy 01 , -(C2-C6)alkenyl-Cy 01 , -(C2-C6)alkynyl-Cy 01 , -O-(C1-C6) alkyl-R 012 , -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ', or -SO2-(C1-C6) alkyl; Or pair (R 06 , R 07 ), when fused with two adjacent carbon atoms, together with the carbon atoms to which they are attached, form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring can be optionally fused with a linear or branched (C1-C6) alkyl group, -NR 013 R 013 ', -(C0-C6) alkyl-Cy 01 or optionally substituted with oxo, W0 is a -CH2- group, a -NH- group or an oxygen atom; R 08 is a hydrogen atom, a linear or branched (C1-C8) alkyl group, -CHR 0a R 0b a group, an aryl group, a heteroaryl group, an aryl(C1-C6)alkyl group, or a heteroaryl(C1-C6)alkyl group, R 09 is a hydrogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, -Cy 02 , -(C1-C6) alkyl-Cy 02 , -(C2-C6)alkenyl-Cy 02 , -(C2-C6)alkynyl-Cy 02 , -Cy 02 -Cy 03 , -(C2-C6)alkynyl-O-Cy 02 , -Cy 02 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 03 , halogen atoms, cyano groups, -C(O)-R 014 or -C(O)-NR 014 R 014 ' and R 010 is a hydrogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, an aryl (C1-C6) alkyl group, a (C1-C6) cycloalkyl alkyl group, a linear or branched (C1-C6) haloalkyl group, or -(C1-C6) alkyl-O-Cy 04 and Or pair (R 09 , R 010 ), when fused with two adjacent carbon atoms, together with the carbon atoms to which they are attached, form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains 1 to 3 heteroatoms selected from O, S and N; R 011 and R 011' are each independently a hydrogen atom, an optionally substituted linear or branched (C1-C6) alkyl group, or -(C0-C6) alkyl-Cy 01 and Or pair (R 011 , R 011 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the N atom is optionally substituted with one or two groups selected from linear or branched (C1-C6) alkyl groups, wherein one or more of the carbon atoms of the linear or branched (C1-C6) alkyl group are optionally deuterated; R 012 -Cy 05 , -Cy 05 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 06 , -Cy 05 -(C0-C6) alkyl-Cy 06 , -Cy 05 -(C0-C6) alkyl-NR 011 -(C0-C6) alkyl-Cy 06 , -Cy 05 -Cy 06 -O-(C0-C6) alkyl-Cy 07 , -Cy 05 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 09 , -Cy 05 -(C0-C6) alkyl-Cy 09 , -NH-C(O)-NH-R 011 , -Cy 05 -(C0-C6) alkyl-NR 011 -(C0-C6) alkyl-Cy 09 , -C(O)-NR 011 R 011 ', -NR 011 R 011 ', -OR 011 , -NR 011 -C(O)-R 011', -O-(C1-C6) alkyl-OR 011 , -SO2-R 011 , -C(O)-OR 011 and R 013 , R 013 ', R 014 and R 014 ' are each independently a hydrogen atom or an optionally substituted linear or branched (C1-C6) alkyl group; R 0a is a hydrogen atom or a linear or branched (C1-C6) alkyl group, R 0b is -OC(O)-OR 0c group, -OC(O)-NR 0c R 0c ' group or -OP(O)(OR 0c )2 units, R 0c and R 0c ' are each independently a hydrogen atom, a linear or branched (C1-C8) alkyl group, a cycloalkyl group, a (C1-C6) alkoxy(C1-C6) alkyl group or a (C1-C6) alkoxycarbonyl(C1-C6) alkyl group, Or pair (R 0c , R 0c ') together with the nitrogen atom to which they are attached form a non-aromatic ring consisting of 5 to 7 ring members which, in addition to the nitrogen atom, may contain 1 to 3 heteroatoms selected from oxygen and nitrogen, wherein the nitrogen is optionally substituted with a linear or branched (C1-C6) alkyl group; Cy 01 , Cy 02 , Cy 03 , Cy 04 , Cy 05 , Cy 06 , Cy 07 , Cy 08 and Cy 010 are, independently of each other, an optionally substituted cycloalkyl group, an optionally substituted heterocycloalkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group; Cy09 teeth

[0489] [ka] and or Cy 09 is -OP(O)(OR 020 )2;-OP(O)(O - M + )2;-(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 ;Hydroxy;Hydroxy(C1-C6)alkyl;-(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl; and -U0-(CH2) q0 -NR 021 R 021 ', and R 015 is a hydrogen atom; -(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 Group; Linear or branched (C1-C6) alkoxy (C1-C6) alkyl group; -U0-(CH2) q0 -NR 021 R 021 ' group; or -(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl group, R 016 represents a hydrogen atom; a hydroxy group; a hydroxy(C1-C6) alkyl group; -(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl group; (CH2) r0 -U0-V0-OP(O)(OR 020 ) 2 groups;-OP(O)(O - M + )2 groups;-OS(O)2OR 020 Group;-S(O)2OR 020 Group;-(CH2) p0-O-(CHR 018 -CHR 019 -O) q0 -R 020 Group;-(CH2) p0 -OC(O)-NR 022 R 023 group; or -U0-(CH2) q0 -NR 021 R 021 'Based on R 017 is a hydrogen atom; -(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 Group: -CH2-P(O)(OR 020 ) 2 units, -OP(O)(OR 020 ) 2 groups;-OP(O)(O - M + )2 group; hydroxy group; hydroxy(C1-C6) alkyl group; -(CH2) r0 -U0-(CH2) s0 -heterocycloalkyl group; -U0-(CH2) q0 -NR 021 R 021 ' group; or aldonic acid, M + is a pharmaceutically acceptable monovalent cation, U0 is a bond or an oxygen atom, V0 is -(CH2) s0 - group or -C(O)- group, R 018 is a hydrogen atom or a (C1-C6)alkoxy(C1-C6)alkyl group, R 019 is a hydrogen atom or a hydroxy(C1-C6)alkyl group, R 020 is a hydrogen atom or a linear or branched (C1-C6) alkyl group, R 021 and R 021 ' are each independently a hydrogen atom, a linear or branched (C1-C6) alkyl group, or a hydroxy(C1-C6) alkyl group, Or pair (R021 , R 021 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring is optionally substituted with hydrogen atoms or linear or branched (C1-C6) alkyl groups; R 022 represents a (C1-C6) alkoxy(C1-C6) alkyl group, -(CH2) p0 -NR 024 R 024 ' group or -(CH2) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 It is the basis, R 023 is a hydrogen atom or a (C1-C6)alkoxy(C1-C6)alkyl group, Or pair (R 022 , R 023 ) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 18 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 5 heteroatoms selected from O, S and N, wherein the resulting ring is optionally substituted with a hydrogen atom, a linear or branched (C1-C6) alkyl group or a heterocycloalkyl group; R 024 and R 024 ' are each independently a hydrogen atom or a linear or branched (C1-C6) alkyl group, Or pair (R 024 , R 024 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring consisting of 5 to 7 ring members, which may contain, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring is optionally substituted with hydrogen atoms or linear or branched (C1-C6) alkyl groups; R 025 is a hydrogen atom, a hydroxy group, or a hydroxy(C1-C6)alkyl group, R 026 is a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, or a cyano group, R 027 is a hydrogen atom or a linear or branched (C1-C6) alkyl group, R 028 is -OP(O)(O - )(O - ) group, -OP(O)(O - )(OR 030 ) group, -OP(O)(OR 030 )(OR 030 ') group, -(CH2) p0 -O-SO2- group, -(CH2) p0 -SO2-O - Group, -(CH2) p0 -O-SO2-OR 030 Group, -Cy 010 , -(CH2) p0 -SO2-OR 030 group, -OC(O)-R 029 The group -OC(O)-OR 029 group or -OC(O)-NR 029 R 029 'Based on R 029 and R 029 ' are each independently a hydrogen atom, a linear or branched (C1-C6) alkyl group or a linear or branched amino(C1-C6) alkyl group, R 030 and R 030 ' are each independently a hydrogen atom, a linear or branched (C1-C6) alkyl group or an aryl (C1-C6) alkyl group, R 031 teeth

[0490] [ka] wherein the ammonium ion is optionally present in zwitterionic form or has a monovalent anionic counterion; n0 is an integer equal to 0 or 1, p0 is an integer equal to 0, 1, 2, or 3; q0 is an integer equal to 1, 2, 3, or 4; r0 and s0 are independently integers equal to 0 or 1; Here, at most, R 03 Group, R 09 group or R 012 one of the groups, if present, is covalently attached to the linker; where the valency of the atom is not exceeded by one or more substituents attached to it. or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0491] In some embodiments, the drug moiety of the present disclosure has formula (II):

[0492] [ka] (In the formula, Z0 is a nitrogen atom or CR 04 It is the basis, R 01 is a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl group, a hydroxy group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, -Cy 08 , -NR 011 R 011 ' and R 02 , R 03 and R 04are each independently a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl, a hydroxy group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, or a -(C0-C6) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C0-C6) alkyl-Cy 01 , -(C2-C6)alkenyl-Cy 01 , -(C2-C6)alkynyl-Cy 01 , -O-(C1-C6) alkyl-NR 011 R 011 ', -O-(C1-C6)alkyl-R 031 , -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ',or -SO2-(C1-C6) alkyl, Or pair (R 02 , R 03 ) or (R 03 , R 04 ) together with the carbon atoms to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, wherein the ring is selected from linear or branched (C1-C6) alkyl, -NR 013 R 013 ', -(C0-C6) alkyl-Cy 01 and oxo, R 06 and R 07 are each independently a hydrogen atom, a halogen atom, a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, a linear or branched (C1-C6) haloalkyl, a hydroxy group, a linear or branched (C1-C6) alkoxy group, an -S-(C1-C6) alkyl group, a cyano group, a nitro group, or a -(C0-C6) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C0-C6) alkyl-Cy 01 , -(C2-C6)alkenyl-Cy 01 , -(C2-C6)alkynyl-Cy 01 , -O-(C1-C6) alkyl-R 012 , -C(O)-OR 011 , -OC(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C1-C6) alkyl-NR 011 -C(O)-R 011 ', -SO2-NR 011 R 011 ', or -SO2-(C1-C6) alkyl; Or pair (R 06 , R 07 ), when fused with two adjacent carbon atoms, together with the carbon atoms to which they are attached, form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, wherein the resulting ring can be optionally fused with a linear or branched (C1-C6) alkyl group, -NR 013 R 013 ', -(C0-C6) alkyl-Cy 01 and oxo, R 08is a hydrogen atom, a linear or branched (C1-C8) alkyl group, an aryl group, a heteroaryl group, an aryl-(C1-C6) alkyl group or a heteroaryl-(C1-C6) alkyl group, R 09 is a linear or branched (C1-C6) alkyl group, a linear or branched (C2-C6) alkenyl group, a linear or branched (C2-C6) alkynyl group, -Cy 02 , -(C1-C6) alkyl-Cy 02 , -(C2-C6)alkenyl-Cy 02 , -(C2-C6)alkynyl-Cy 02 , -Cy 02 -Cy 03 , -(C2-C6)alkynyl-O-Cy 02 , -Cy 02 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy 03 , halogen atoms, cyano groups, -C(O)-R 014 , -C(O)-NR 014 R 014 ' and R 011 and R 011 ' are each independently a hydrogen atom, an optionally substituted linear or branched (C1-C6) alkyl group, or -(C0-C6) alkyl-Cy 01 and Or pair (R 011 , R 011 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the N atom is optionally substituted with a linear or branched (C1-C6) alkyl group, and wherein one or more of the carbon atoms of the linear or branched (C1-C6) alkyl group is optionally deuterated; R 012 -Cy 05 , -Cy 05 -(C0-C6) alkyl-Cy 06 , -Cy 05 -(C0-C6)alkyl-O-(C0-C6)alkyl-Cy06 , -Cy 05 -(C0-C6) alkyl-NR 011 -(C0-C6) alkyl-Cy 06 , -Cy 05 -Cy 06 -O-(C0-C6) alkyl-Cy 07 , -Cy 05 -(C0-C6) alkyl-Cy 09 , -NH-C(O)-NH-R 011 , -C(O)-NR 011 R 011 ', -NR 011 R 011 ', -OR 011 , -NR 011 -C(O)-R 011 ', -O-(C1-C6) alkyl-OR 011 , -SO2-R 011 , or -C(O)-OR 011 and R 013 , R 013 ', R 014 and R 014 ' are each independently a hydrogen atom or an optionally substituted linear or branched (C1-C6) alkyl group; Cy 01 , Cy 02 , Cy 03 , Cy 05 , Cy 06 , Cy 07 and Cy 08 are, independently of each other, an optionally substituted cycloalkyl group, an optionally substituted heterocycloalkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group; Cy 09 teeth

[0493] [ka] and where R 015 , R 016 and R 017is as defined for formula (I), R 031 teeth

[0494] [ka] where R 027 and R 028 is as defined for formula (I), Here, at most, R 03 Group, R 09 group or R 012 One of the groups, if present, is covalently attached to the linker or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0495] In some embodiments, the drug moiety of the present disclosure has formula (III):

[0496] [ka] (In the formula, R 01 is a linear or branched (C1-C6) alkyl group, R 03 is -O-(C1-C6) alkyl-NR 011 R 011 'or

[0497] [ka] and where R 011 and R 011 ' are each independently a hydrogen atom, an optionally substituted linear or branched (C1-C6) alkyl group, or -(C0-C6) alkyl-Cy 01 and Or pair (R 011 , R 011') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to the nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein the N atom may be substituted with one or two groups selected from a hydrogen atom or a linear or branched (C1-C6) alkyl group; where R 027 is a hydrogen atom, and R 028 is -(CH2) p0 -O-SO2-O - group or -(CH2) p0 -SO2-OR 030 It is the basis, R 09 is a linear or branched (C2-C6) alkynyl group or -Cy 02 and R 012 -Cy 05 , -Cy 05 -(C0-C6) alkyl-Cy 06 or -Cy 05 -(C0-C6) alkyl-Cy 09 and Cy 01 , Cy 02 , Cy 05 and Cy 06 are, independently of each other, a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group, each of which is optionally substituted; Cy 09 teeth,

[0498] [ka] and p0, R 015 , R 016 and R 017 is as defined for formula (I), Here, at most, R 03 Group, R 09 group or R 012 One of the groups, if present, is covalently attached to the linker or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0499] In some embodiments, Cy 01 , Cy 02 , Cy 03 , Cy 04 , Cy 05 , Cy 06 , Cy 07 , Cy 08 and Cy 010 are each independently an optionally substituted cycloalkyl group, an optionally substituted heterocycloalkyl group, an optionally substituted aryl group or an optionally substituted heteroaryl group, wherein the optional substituents are optionally substituted linear or branched (C1-C6) alkyl, optionally substituted linear or branched (C2-C6) alkenyl group, optionally substituted linear or branched (C2-C6) alkynyl group, optionally substituted linear or branched (C1-C6) alkoxy, optionally substituted (C1-C6) alkyl-S-, hydro and R is selected from oxy, oxo (or N-oxide, where appropriate), nitro, cyano, —C(O)—OR′, —OC(O)—R′, —C(O)—NR′R′, —NR′R′, —(C═NR′)—OR′, linear or branched (C1-C6) haloalkyl, trifluoromethoxy, or halogen, where R′ and R′ are each independently a hydrogen atom or an optionally substituted linear or branched (C1-C6) alkyl group, and where one or more of the carbon atoms of the linear or branched (C1-C6) alkyl group is optionally deuterated.

[0500] In some embodiments, the drug moiety (D) is

[0501] [ka]

[0502] [ka] or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0503] Additionally, the drug moiety of the present disclosure may be:

[0504] [ka]

[0505] [ka] It may include any one of the following:

[0506] In some embodiments, the linker-drug (or "linker-payload") moiety (LD) may comprise a compound selected from Table A.

[0507] Drug Load The drug load is represented by p and is also referred to herein as the drug-to-antibody ratio (DAR). The drug load may range from 1 to 16 per drug moiety antibody or antigen-binding fragment. In some embodiments, p is an integer from 1 to 16. In some embodiments, p is an integer from 1 to 16, 1 to 15, 1 to 14, 1 to 13, 1 to 12, 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2. In some embodiments, p is an integer from 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, or 2 to 3. In some embodiments, p is an integer from 1 to 16. In some embodiments, p is an integer from 1 to 8. In some embodiments, p is an integer from 1 to 5. In some embodiments, p is an integer from 2 to 4. In some embodiments, p is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, p is 2. In some embodiments, p is 4.

[0508] Drug loading may be limited by the number of binding sites on the antibody or antigen-binding fragment. In some embodiments, the linker moiety (L) of the ADC is attached to the antibody or antigen-binding fragment via a chemically active group on one or more amino acid residues on the antibody or antigen-binding fragment. For example, the linker may be attached to the antibody or antigen-binding fragment via a free amino, imino, hydroxyl, thiol, or carboxyl group (e.g., to the N- or C-terminus, to the epsilon-amino group of one or more lysine residues, to the free carboxylic acid group of one or more glutamic acid or aspartic acid residues, or to the sulfhydryl group of one or more cysteine ​​residues). The linker attachment site may be a natural residue in the amino acid sequence of the antibody or antigen-binding fragment, or may be introduced into the antibody or antigen-binding fragment by, for example, recombinant DNA techniques (e.g., by introducing a cysteine ​​residue into the amino acid sequence) or protein biochemistry (e.g., by reduction, pH adjustment, or hydrolysis).

[0509] In some embodiments, the number of drug moieties that can be conjugated to an antibody or antigen-binding fragment is limited by the number of free cysteine ​​residues. For example, if the bond is a cysteine ​​thiol group, the antibody may have only one or a few cysteine ​​thiol groups, or only one or a few sufficiently reactive thiol groups to which a linker can be attached. Generally, antibodies do not contain many free and reactive cysteine ​​thiol groups that can be linked to a drug moiety. In fact, most cysteine ​​thiol residues in antibodies are involved in either inter- or intra-chain disulfide bonds. As a result, in some embodiments, conjugation to cysteines may require at least partial reduction of the antibody. Excessive conjugation of linker-toxins to an antibody may compromise antibody stability by reducing the number of cysteine ​​residues available for disulfide bond formation. Consequently, an optimal drug:antibody ratio should increase the potency of the ADC (by increasing the number of drug moieties conjugated per antibody) without compromising the stability of the antibody or antigen-binding fragment. In some embodiments, the optimal ratio may be 2, 4, 6, or 8. In some embodiments, the optimal ratio may be 2 or 4.

[0510] In some embodiments, the antibody or antigen-binding fragment is exposed to reducing conditions prior to conjugation to generate one or more free cysteine ​​residues. In some embodiments, the antibody may be reduced under partial or total reducing conditions using a reducing agent, such as dithiothreitol (DTT) or tris(2-carboxyethyl)phosphine (TCEP), to generate reactive cysteine ​​thiol groups. Unpaired cysteines may be generated through partial reduction using a limited molar equivalent of TCEP, which reduces the interchain disulfide bonds connecting the light and heavy chains (one pair per HL pair) and the two heavy chains in the hinge region (two pairs per HH pair in the case of human IgG1), while leaving the intrachain disulfide bonds intact (Stefano et al. (2013) Methods Mol Biol. 1045:145-71). In embodiments, disulfide bonds within the antibody are electrochemically reduced, for example, by using a working electrode that alternately applies a reducing and oxidizing voltage. This approach can allow online coupling of disulfide bond reduction to an analytical device (e.g., an electrochemical detection device, an NMR spectrometer, or a mass spectrometer) or a chemical separation device (e.g., a liquid chromatography (e.g., HPLC) or an electrophoresis device (see, e.g., U.S. Patent Application Publication No. 2014 / 0069822)). In some embodiments, the antibody is subjected to denaturing conditions to reveal reactive nucleophilic groups on amino acid residues, e.g., cysteines.

[0511] Drug loading of ADCs may be controlled in different ways, for example: (i) by limiting the molar excess of drug-linker intermediate or linker reagent relative to antibody; (ii) by limiting the conjugation reaction time or temperature; (iii) by partial or limited reducing conditions for cysteine ​​thiol modifications; and / or (iv) by recombinantly engineering the amino acid sequence of the antibody to modify the number and position of cysteine ​​residues to control the number and / or position of linker-drug bonds.

[0512] In some embodiments, a free cysteine ​​residue is introduced into the amino acid sequence of an antibody or antigen-binding fragment. For example, a cysteine-engineered antibody may be prepared in which one or more amino acids of a parent antibody are replaced with a cysteine ​​amino acid. Both forms of antibodies may be engineered, i.e., mutated, in this manner. For example, a parent Fab antibody fragment may be engineered to form a cysteine-engineered Fab, referred to as a "ThioFab." Similarly, a parent monoclonal antibody may be engineered to form a "ThioMab." A single-site mutation results in a single engineered cysteine ​​residue in a ThioFab, while a single-site mutation results in two engineered cysteine ​​residues in a ThioMab due to the dimeric nature of IgG antibodies. DNA encoding amino acid sequence variants of a parent polypeptide can be prepared by various methods known in the art (see, for example, the methods described in WO 2006 / 034488). These methods include, but are not limited to, site-directed (or oligonucleotide-mediated) mutagenesis, PCR mutagenesis, and cassette mutagenesis of initially prepared DNA encoding the polypeptide. Recombinant antibody variants may also be constructed by restriction fragment engineering or overlap extension PCR using synthetic oligonucleotides. ADCs of formula (1) include, but are not limited to, antibodies with one, two, three, or four engineered cysteine ​​amino acids (Lyon et al. (2012) Methods Enzymol. 502:123-38). In some embodiments, one or more free cysteine ​​residues are already present in an unengineered antibody or antigen-binding fragment, in which case the existing free cysteine ​​residues may be used to conjugate the antibody or antigen-binding fragment to a drug moiety.

[0513] When more than one nucleophilic group reacts with a drug-linker intermediate or with a linker moiety reagent followed by a drug moiety reagent in a reaction mixture containing multiple copies of an antibody or antigen-binding fragment and a linker moiety, the resulting product can be a mixture of ADC compounds with a distribution of one or more drug moieties attached to each copy of the antibody or antigen-binding fragment in the mixture. In some embodiments, the drug load in the mixture of ADCs resulting from the conjugation reaction ranges from 1 to 16 attached drug moieties per antibody or antigen-binding fragment. The average number of drug moieties per antibody or antigen-binding fragment (i.e., average drug load, or average p) can be calculated by any conventional method known in the art, for example, by mass spectrometry (e.g., liquid chromatography-mass spectrometry (LC-MS)) and / or high-performance liquid chromatography (e.g., HIC-HPLC). In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is determined by liquid chromatography-mass spectrometry (LC-MS). In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 1.5 to about 3.5, about 2.5 to about 4.5, about 3.5 to about 5.5, about 4.5 to about 6.5, about 5.5 to about 7.5, about 6.5 to about 8.5, or about 7.5 to about 9.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 2 to about 4, about 3 to about 5, about 4 to about 6, about 5 to about 7, about 6 to about 8, about 7 to about 9, about 2 to about 8, or about 4 to about 8.

[0514] In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 2. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2, about 2.1, about 2.2, about 2.3, about 2.4, or about 2.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is 2.

[0515] In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 4. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4, about 4.1, about 4.2, about 4.3, about 4.4, or about 4.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is 4.

[0516] In some embodiments, the term "about," when used in reference to the average number of drug moieties per antibody or antigen-binding fragment, means plus or minus 20%, 15%, 10%, 5%, or 1%. In one embodiment, the term "about" refers to a range of values ​​that are 10% greater or less than a particular value. In another embodiment, the term "about" refers to a range of values ​​that are 5% greater or less than a particular value. In another embodiment, the term "about" refers to a range of values ​​that are 1% greater or less than a particular value.

[0517] Individual ADC compounds or "species" may be identified in the mixture by mass spectroscopy and separated, for example, by UPLC or HPLC, such as hydrophobic interaction chromatography (HIC-HPLC). In some embodiments, a homogeneous or near-homogeneous ADC product having a single loading value may be isolated from the conjugation mixture, for example, by electrophoresis or chromatography.

[0518] In some embodiments, high drug loads (e.g., p>16) may result in aggregation, insolubility, toxicity, or loss of cell permeability of certain antibody-drug conjugates. High drug loads may also negatively impact the pharmacokinetics (e.g., clearance) of certain ADCs. In some embodiments, low drug loads (e.g., p<2) may reduce the potency of certain ADCs against target-expressing cells. In some embodiments, the drug load for an ADC of the present disclosure ranges from about 2 to about 16, about 2 to about 10, about 2 to about 8; about 2 to about 6; about 2 to about 5; about 3 to about 5; about 2 to about 4; or about 4 to about 8.

[0519] In some embodiments, a drug load and / or average drug load of about 2 is achieved, for example, using partial reduction of intrachain disulfides in the antibody or antigen-binding fragment, to provide beneficial properties. In some embodiments, a drug load and / or average drug load of about 4, about 6, or about 8 is achieved, for example, using partial reduction of intrachain disulfides in the antibody or antigen-binding fragment, to provide beneficial properties. In some embodiments, a drug load and / or average drug load of less than about 2 results in an unacceptably high level of unconjugated antibody species, which may compete with the ADC for binding to the target antigen CD74 and / or may reduce the efficacy of treatment. In some embodiments, a drug load and / or average drug load of more than about 16 may result in an unacceptably high level of product heterogeneity and / or ADC aggregation. A drug load and / or average drug load of more than about 16 may also affect the stability of the ADC, due to loss of one or more chemical bonds required to stabilize the antibody or antigen-binding fragment.

[0520] The present disclosure includes methods for producing the described ADCs. Briefly, an ADC comprises an antibody or antigen-binding fragment that remains as an antibody or antigen-binding fragment, a drug moiety (e.g., an Mcl-1 inhibitor), and a linker connecting the drug moiety and the antibody or antigen-binding fragment. In some embodiments, an ADC may be prepared using a linker with a reactive functional group for covalently attaching the drug moiety to the antibody or antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment is functionalized to prepare a functional group that is reactive with a linker or drug-linker intermediate. For example, in some embodiments, a cysteine ​​thiol of the antibody or antigen-binding fragment can form a bond with a reactive functional group of a linker or drug-linker intermediate to produce an ADC. In some embodiments, antibodies or antigen-binding fragments are prepared using bacterial transglutaminase (BTG), a reactive glutamine specifically functionalized with an amine-containing cyclooctyne BCN (N-[(1R,8S,9s)-bicyclo[6.1.0]non-4-yn-9-ylmethyloxycarbonyl]-1,8-diamino-3,6-dioxaoctane) moiety. In some embodiments, site-specific conjugation of a linker or drug-linker intermediate to the BCN moiety of the antibody or antigen-binding fragment is performed, for example, as described and exemplified herein. Generation of ADCs can be achieved by techniques known to those skilled in the art.

[0521] In some embodiments, ADCs are produced by contacting an antibody or antigen-binding fragment with a linker and a drug moiety (e.g., an Mcl-1 inhibitor) in a sequential manner, such that the antibody or antigen-binding fragment is first covalently linked to the linker and then a preformed antibody-linker intermediate is reacted with the drug moiety. The antibody-linker intermediate may or may not undergo a purification step before contacting the drug moiety. In other embodiments, ADCs are produced by contacting an antibody or antigen-binding fragment with a linker-drug compound that is preformed by reacting the linker with the drug moiety. The preformed linker-drug compound may or may not undergo a purification step before contacting the antibody or antigen-binding fragment. In other embodiments, the antibody or antigen-binding fragment is contacted with the linker and drug moiety in a single reaction mixture, allowing for simultaneous formation of covalent bonds between the antibody or antigen-binding fragment and the linker and between the linker and the drug moiety. This method of producing an ADC may involve contacting an antibody or antigen-binding fragment with an antibody or antigen-binding fragment and then adding a linker to the reaction mixture, or vice versa. In some embodiments, the ADC is produced by reacting an antibody or antigen-binding fragment with a linker linked to a drug moiety, e.g., an Mcl-1 inhibitor, under conditions that allow conjugation.

[0522] The ADC prepared according to the above-mentioned method may be subjected to a purification step. The purification step may involve any biochemical method known in the art for purifying proteins, or any combination of these methods. These include, but are not limited to, tangential flow filtration (TFF), affinity chromatography, ion exchange chromatography, any charge- or isoelectric-based chromatography, mixed-mode chromatography, such as CHT (ceramic hydroxyapatite), hydrophobic interaction chromatography, size exclusion chromatography, dialysis, filtration, selective precipitation, or any combination thereof.

[0523] Therapeutic Uses and Compositions Disclosed herein are methods for using compositions described herein, for example, the disclosed ADC compounds and compositions, in the treatment of subjects with disorders, for example, cancer.Compositions, for example, ADCs, can be administered alone or in combination with at least one additional inactive agent and / or active agent, for example, at least one additional therapeutic agent, and can be administered in any pharmaceutically acceptable formulation, dosage, and dosing regimen.The effectiveness of treatment can be evaluated based on toxicity and efficacy indicators and adjusted accordingly.Measures of effectiveness include, but are not limited to, cytostatic and / or cytotoxic effects observed in vitro or in vivo, reduction in tumor volume, tumor growth inhibition, and / or prolonged survival.

[0524] Methods for determining whether an ADC exerts a cytostatic and / or cytotoxic effect on cells are known. For example, the cytotoxicity or cytostatic activity of an ADC can be measured by, for example, exposing mammalian cells expressing the target antigen CD74 of the ADC in cell culture medium; culturing the cells for a period of about 6 hours to about 6 days; and measuring cell viability (for example, using CellTiter-Glo® (CTG) or MTT cell viability assay). Cell-based in vitro assays can also be used to measure viability (proliferation), cytotoxicity, and induction of apoptosis (caspase activation) of the ADC.

[0525] To assess cytotoxicity, necrosis or apoptosis (programmed cell death) may be measured. Necrosis is typically accompanied by increased plasma membrane permeability, cell swelling, and plasma membrane rupture. Apoptosis can be quantified, for example, by measuring DNA fragmentation. Commercially available photometric methods for quantitatively assessing DNA fragmentation in vitro are available. Examples of such assays include TUNEL (detecting the incorporation of labeled nucleotides into fragmented DNA) and ELISA-based assays, which are described in Biochemica (1999) 2:34-7 (Roche Molecular Biochemicals).

[0526] Apoptosis may also be determined by measuring morphological changes in cells. For example, loss of plasma membrane integrity, similar to necrosis, may be determined by measuring the uptake of certain dyes (e.g., fluorescent dyes such as acridine orange or ethidium bromide). Methods for measuring the number of apoptotic cells are described by Duke and Cohen in Current Protocols in Immunology (Coligan et al., eds. (1992) pp. 3.17.1-3.17.16). Cells may also be labeled with DNA dyes (e.g., acridine orange, ethidium bromide, or propidium iodide) and observed for chromatin condensation and margination along the inner side of the nuclear membrane. Apoptosis may also be determined, in some embodiments, by screening for caspase activity. In some embodiments, the Caspase-Glo® assay may be used to measure the activity of caspase-3 and caspase-7. In some embodiments, the assay provides a luminescent caspase-3 / 7 substrate in a reagent optimized for caspase activity, luciferase activity, and cell lysis. In some embodiments, addition of the Caspase-Glo® 3 / 7 Reagent in an "add-mix-measure" format can result in cell lysis, followed by caspase cleavage of the substrate and the generation of a "glow-type" luminescent signal generated by luciferase. In some embodiments, luminescence can be proportional to the amount of caspase activity present and can serve as an indicator of apoptosis. Other morphological changes that can be measured to determine apoptosis include, for example, cytoplasmic condensation, increased membrane blebbing, and cell shrinkage. Determining any of these effects on cancer cells indicates that the ADC is useful in treating cancer.

[0527] Cell viability may be measured, for example, by determining cellular uptake of a dye, such as neutral red, trypan blue, crystal violet, or ALAMAR™ blue (see, e.g., Page et al. (1993) Intl J Oncology 3:473-6). In such assays, cells are incubated in medium containing the dye, washed, and the remaining dye, which reflects cellular uptake of the dye, is measured spectrophotometrically.

[0528] Cell viability may also be measured, for example, by quantifying ATP, an indicator of metabolically active cells. In some embodiments, the in vitro potency and / or cell viability of the prepared ADC or Mcl-1 inhibitor compound may be assessed using the CellTiter-Glo® (CTG) cell viability assay, as described in the examples provided herein. In this assay, in some embodiments, a single reagent (CellTiter-Glo® Reagent) is added directly to cells cultured in serum-supplemented medium. Addition of the reagent results in cell lysis and the generation of a luminescent signal proportional to the amount of ATP present. The amount of ATP is directly proportional to the number of cells present in the culture.

[0529] Cell viability may also be measured, for example, by measuring the reduction of tetrazolium salts. In some embodiments, the in vitro efficacy and / or cell viability of the prepared ADC or Mcl-1 inhibitor compound may be evaluated using an MTT cell viability assay, as described in the examples provided herein. In this assay, in some embodiments, the yellow tetrazolium MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) is reduced by metabolically active cells, in part by the action of dehydrogenase enzymes, to generate reducing equivalents, such as NADH and NADPH. The resulting intracellular purple formazan may then be solubilized and quantified by spectrophotometric means.

[0530] In certain aspects, the present disclosure features methods for killing cancer cells or tissues and inhibiting or modulating their growth by disrupting the expression and / or activity of Mcl-1 and / or one or more upstream modulators or downstream targets thereof. The methods may be used in any subject for whom disruption of Mcl-1 expression and / or activity provides a therapeutic benefit. Subjects that may benefit from disrupting Mcl-1 expression and / or activity include, but are not limited to, those with or at risk of having cancer, such as tumors or blood cancers. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[0531] An exemplary method includes contacting cells with an ADC, as described herein, in an effective amount, i.e., an amount sufficient to kill the cells. The method may be used on cells in culture, e.g., in vitro, in vivo, ex vivo, or in situ. For example, CD74-expressing cells (e.g., cells collected by biopsy of a tumor or metastatic lesion; cells from an established cancer cell line; or recombinant cells) may be cultured in vitro in culture medium, and the contacting step may be affected by adding an ADC to the culture medium. The method will result in the killing of cells expressing CD74, including specifically CD74-expressing cancer cells. Alternatively, the ADC may be administered to a subject by any suitable route of administration (e.g., intravenously, subcutaneously, or by direct contact with tumor tissue) to have an in vivo effect.

[0532] The in vivo efficacy of the disclosed ADC therapeutic compositions may be evaluated in a suitable animal model. For example, a xenogeneic cancer model may be used, in which cancer explants or serial xenografts are introduced into immunodeficient animals, such as nude or SCID mice (Klein et al. (1997) Nature Med. 3:402-8). Efficacy may be predicted using assays that measure tumor formation, tumor regression, or inhibition of metastasis, etc.

[0533] In vivo assays that evaluate the mechanism of tumor killing promotion, such as apoptosis, may also be used. In some embodiments, xenografts from tumor-bearing mice treated with therapeutic compositions may be examined for the presence of apoptotic foci and compared with untreated control xenograft-bearing mice. The degree of apoptotic foci observed in the tumors of treated mice provides an indication of the therapeutic effectiveness of the composition.

[0534] Further provided herein is a method for treating disorders, for example, cancer.The compositions described herein, for example, the ADCs disclosed herein, can be administered to non-human mammals or human subjects for therapeutic purposes.The therapeutic method includes administering a therapeutically effective amount of a composition comprising an Mcl-1 inhibitor, for example, an ADC, to a subject who has or is suspected of having cancer, wherein the inhibitor is linked to a targeting antibody that binds to an antigen, and the antigen (1) is expressed on cancer cells, (2) is accessible for binding, and / or (3) is located or primarily expressed on the surface of cancer cells compared to non-cancer cells.

[0535] An exemplary embodiment is a method of treating a subject having or suspected of having cancer, comprising administering to the subject a therapeutically effective amount of a composition, e.g., an ADC, composition, or pharmaceutical composition disclosed herein (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein). In some embodiments, the cancer expresses the target antigen CD74. In some embodiments, the cancer is a tumor or blood cancer. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[0536] Another exemplary embodiment is a method of delivering an Mcl-1 inhibitor to a cell expressing CD74, comprising conjugating the Mcl-1 inhibitor to an antibody that immunospecifically binds to a CD74 epitope and exposing the cell to the ADC. Exemplary cancer cells expressing CD74 for which the ADCs of the present disclosure are applicable include multiple myeloma cells.

[0537] In certain aspects, the present disclosure further provides methods of reducing or inhibiting tumor (e.g., CD74-expressing tumor) growth, comprising administering a therapeutically effective amount of an ADC or a composition comprising an ADC. In some embodiments, the treatment is sufficient to reduce or inhibit tumor growth, reduce the number or size of metastatic lesions, reduce tumor burden, reduce primary tumor burden, reduce invasiveness, prolong survival, and / or maintain or improve quality of life in a patient. In some embodiments, the tumor is resistant or refractory to treatment with an antibody or antigen-binding fragment of the ADC (e.g., an anti-CD74 antibody) when administered alone, and / or the tumor is resistant or refractory to treatment with an Mcl-1 inhibitor drug moiety when administered alone.

[0538] An exemplary embodiment is a method of reducing or inhibiting tumor growth in a subject, comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein). In some embodiments, the tumor expresses the target antigen CD74. In some embodiments, the tumor is breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular carcinoma, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the tumor is gastric cancer. In some embodiments, administration of the ADC, composition, or pharmaceutical composition reduces or inhibits tumor growth by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99% compared to growth in the absence of treatment.

[0539] Another exemplary embodiment is a method of delaying or slowing tumor growth in a subject, comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein). In some embodiments, the tumor expresses the target antigen CD74. In some embodiments, the tumor is breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular carcinoma, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the tumor is gastric cancer. In some embodiments, administration of the ADC, composition, or pharmaceutical composition delays or slows tumor growth by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99% compared to growth in the absence of treatment.

[0540] In certain aspects, the present disclosure further provides methods of reducing or slowing the expansion of a cancer cell population (e.g., a CD74-expressing cancer cell population), comprising administering a therapeutically effective amount of an ADC or a composition comprising an ADC.

[0541] An exemplary embodiment is a method of reducing or slowing the expansion of a cancer cell population in a subject, comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein). In some embodiments, the cancer cell population expresses the target antigen CD74. In some embodiments, the cancer cell population is derived from a tumor or hematological cancer. In some embodiments, the cancer cell population is derived from breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies of T-cell or B-cell origin, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. In some embodiments, the cancer cell population is derived from lymphoma or gastric cancer. In some embodiments, administration of the ADC, composition, or pharmaceutical composition reduces the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99% compared to the population in the absence of treatment. In some embodiments, administration of the ADC, composition, or pharmaceutical composition slows the expansion of the cancer cell population by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99% compared to expansion in the absence of treatment.

[0542] Also provided herein are methods for determining whether a subject with or suspected of having cancer will respond to treatment with the disclosed ADCs and compositions. An exemplary embodiment is a method for determining whether a subject with or suspected of having cancer will respond to treatment with an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein) by providing a biological sample from the subject; contacting the sample with an ADC; and detecting binding of the ADC to cancer cells in the sample. In some embodiments, the sample is a tissue biopsy sample, a blood sample, or a bone marrow sample. In some embodiments, the method includes providing a biological sample from the subject; contacting the sample with an ADC; and detecting one or more markers of cancer cell death in the sample (e.g., increased expression of one or more apoptosis markers, decreased expansion of a cancer cell population in culture, etc.).

[0543] Therapeutic uses of the disclosed ADCs and compositions are further provided herein. An exemplary embodiment is an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein) for use in treating a subject having or suspected of having cancer (e.g., a CD74-expressing cancer). Another exemplary embodiment is the use of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein) in treating a subject having or suspected of having cancer (e.g., a CD74-expressing cancer). Another exemplary embodiment is the use of an ADC, composition, or pharmaceutical composition (e.g., any of the exemplary ADCs, compositions, or pharmaceutical compositions disclosed herein) in a method of manufacturing a medicament for treating a subject having or suspected of having cancer (e.g., a CD74-expressing cancer). Methods for identifying subjects with cancers that express the target antigen CD74 are known in the art and may be used to identify patients suitable for treatment with the disclosed ADC compounds or compositions.

[0544] Additionally, the ADCs of the present disclosure may be administered to non-human mammals expressing antigens to which the ADCs can bind, for veterinary purposes or as animal models of human disease. Regarding the latter, such animal models can be useful for evaluating the therapeutic efficacy of the disclosed ADCs (e.g., testing dosages and time courses of administration).

[0545] Therapeutic compositions used in practicing the aforementioned methods may be formulated into pharmaceutical compositions comprising a pharmaceutically acceptable carrier suitable for the desired delivery method. An exemplary embodiment is a pharmaceutical composition comprising an ADC of the present disclosure and a pharmaceutically acceptable carrier, e.g., suitable for a selected means of administration, e.g., intravenous administration. The pharmaceutical composition may also include one or more additional inactive and / or therapeutic agents suitable for treating or preventing cancer (e.g., standard of care agents, etc.). The pharmaceutical composition may also include one or more carrier, excipient, and / or stabilizer components, etc. Methods for formulating such pharmaceutical compositions and suitable formulations are known in the art (see, e.g., "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA).

[0546] Suitable carriers include any material that, when combined with the therapeutic composition, maintains the anti-tumor function of the therapeutic composition and is generally non-reactive with the patient's immune system. Pharmaceutically acceptable carriers include any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonicity and absorption delaying agents, etc. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, mesylate, etc., and combinations thereof. In many cases, isotonicity agents, such as sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride, are included in the composition. Pharmaceutically acceptable carriers may also contain small amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives, or buffers, that enhance the shelf life or effectiveness of ADCs.

[0547] The pharmaceutical compositions of the present disclosure may be administered by various methods known in the art. The route and / or mode of administration may vary depending on the desired results. In some embodiments, the therapeutic formulation is solubilized and administered via any route capable of delivering the therapeutic composition to the cancer site. Potentially effective routes of administration include, but are not limited to, parenteral (e.g., intravenous, subcutaneous), intraperitoneal, intramuscular, intratumoral, intradermal, intraorgan, orthotopic, etc. In some embodiments, administration is intravenous, subcutaneous, intraperitoneal, or intramuscular. The pharmaceutically acceptable carrier should be suitable for the route of administration, e.g., intravenous or subcutaneous administration (e.g., by injection or infusion). Depending on the route of administration, the active compound, i.e., the ADC and / or any additional therapeutic agent, may be coated with a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound. Administration may be systemic or local.

[0548] The therapeutic compositions disclosed herein may be sterile and stable under the conditions of manufacture and storage and may be in a variety of forms. These include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. The form depends on the intended mode of administration and therapeutic application. In some embodiments, the disclosed ADCs may be incorporated into pharmaceutical compositions suitable for parenteral administration. Injectable solutions may consist of either liquid or lyophilized dosage forms in flint or amber vials, ampoules, or pre-filled syringes, or other known delivery or storage devices. In some embodiments, one or more of the ADCs or pharmaceutical compositions are supplied as a dry, sterile, lyophilized powder or water-free concentrate in a sealed container, which can be reconstituted (e.g., with water or saline) to the appropriate concentration for administration to a subject.

[0549] Typically, the disclosed compositions, for example, the disclosed ADCs, are used in the pharmaceutical compositions of the present disclosure in a therapeutically effective amount or effective amount.Compositions, for example, those comprising ADCs, can be formulated into pharmaceutically acceptable dosage forms by conventional methods known in the art.The dosage and administration protocol for treating cancer using the above-mentioned methods will vary with the method and target cancer, and will generally depend on many other factors recognized in the art.

[0550] Dosage regimens for the compositions disclosed herein, e.g., those comprising an ADC alone or in combination with at least one additional inactive and / or active therapeutic agent, may be adjusted to provide the optimal desired response (e.g., a therapeutic response). For example, a single bolus of one or both agents may be administered all at once, or several divided doses may be administered over a predetermined period of time, or the doses of one or both agents may be proportionally increased or decreased as indicated by the exigencies of the therapeutic situation. In some embodiments, treatment involves single bolus or repeated administration of an ADC preparation via an acceptable route of administration. In some embodiments, the ADC is administered to the patient daily, weekly, monthly, or any period therebetween. For any particular subject, the specific dosing regimen may be adjusted over time according to the individual needs and the professional judgment of the treating clinician. Parenteral compositions may be formulated in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

[0551] Dosage values ​​for compositions containing an ADC and / or any additional therapeutic agents may be selected based on the unique characteristics of the active compound and the particular therapeutic effect to be achieved. A physician or veterinarian may begin dosing of the ADC used in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. Generally, the effective dose of the compositions of the present disclosure, with respect to the treatment of cancer, may vary depending on many different factors, including the means of administration, the target site, the physiological condition of the patient, whether the patient is human or animal, other pharmaceutical agents administered, and whether the treatment is prophylactic or therapeutic. The selected dosage level may also depend on various pharmacokinetic factors, including the activity of the particular composition of the present disclosure or its ester, salt, or amide used, the route of administration, the time of administration, the excretion rate of the particular compound used, the duration of treatment, other drugs, compounds, and / or materials used in combination with the particular composition used, the age, sex, weight, condition, general health, and past medical history of the patient being treated, and similar factors. Treatment dosages may be titrated to optimize safety and efficacy.

[0552] The toxicity and therapeutic efficacy of compounds provided herein can be determined by standard pharmaceutical procedures in cell culture or animal models.For example, LD50, ED50, EC50 and IC50 can be determined, and the dose ratio between toxicity and therapeutic effect (LD50 / ED50) can be calculated as therapeutic index.The data obtained from in vitro and in vivo assays can be used to estimate or formulate the dosage range for use in humans.For example, the compositions and methods disclosed herein can be first evaluated in heterogeneous cancer models (for example, NCI-H929 multiple myeloma mouse model).

[0553] In some embodiments, the ADC or composition comprising the ADC is administered on a single occasion. In other embodiments, the ADC or composition comprising the ADC is administered on multiple occasions. The interval between single doses may be, for example, daily, weekly, monthly, or yearly. The interval may be irregular based on measuring the blood level of the administered drug (e.g., ADC) in the patient to maintain a relatively consistent plasma concentration of the drug. The dosage and frequency of administration of the ADC or composition comprising the ADC may also vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, relatively low dosages may be administered at relatively infrequent intervals over an extended period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, relatively high dosages at relatively short intervals may sometimes be required until the progression of the disease slows or terminates, preferably until the patient shows partial or complete improvement in one or more symptoms of the disease. Thereafter, the patient may be administered a lower, e.g., prophylactic, regimen.

[0554] The above therapeutic approaches may be combined with any one of a variety of additional surgical, chemotherapy, or radiation therapy regimens. In some embodiments, an ADC or composition disclosed herein is co-formulated with and / or co-administered with one or more additional therapeutic agents, e.g., one or more chemotherapeutic agents, one or more standard of care agents for the particular condition being treated.

[0555] Kits for use in the therapeutic and / or diagnostic applications described herein are also provided. Such kits may include a carrier, package, or container, which may be compartmentalized to accommodate one or more containers, e.g., vials, tubes, etc., each containing one of the individual components to be used in the methods disclosed herein. A label may be present on or with the container to indicate that the ADC or composition in the kit is to be used for a particular therapeutic or non-therapeutic application, e.g., prognostic, preventative, diagnostic, or laboratory application. The label may also indicate instructions for either in vivo or in vitro use, e.g., those described herein. Instructions and other information may also be included in an insert or label included with or on the kit. The label may be on or associated with the container. The label may be on the container if letters, numbers, or other characters forming the label are molded or etched into the container itself. The label may also be associated with the container if present in a storage receptacle or carrier that also holds the container, e.g., as a package insert. The label may indicate that the ADC or composition in the kit is used for diagnosing or treating a condition described herein, eg, cancer.

[0556] In some embodiments, the kit comprises an ADC or a composition comprising an ADC. In some embodiments, the kit further comprises one or more additional components, including, but not limited to, instructions for use; other reagents, such as therapeutic agents (e.g., standard of care drugs); devices, containers, or other materials for preparing the ADC for administration; pharmaceutically acceptable carriers; and devices, containers, or other materials for administering the ADC to a subject. The instructions for use may include guidance for therapeutic application, including, for example, recommended dosages and / or modes of administration in patients with or suspected of having cancer. In some embodiments, the kit comprises an ADC and instructions for using the ADC in the treatment, prevention, and / or diagnosis of cancer.

[0557] Combination Therapy In some embodiments, the present disclosure provides methods of treatment in which an antibody-drug conjugate disclosed herein is administered in combination with one or more additional therapeutic agents. Exemplary combination partners are disclosed herein.

[0558] In certain embodiments, the combinations described herein include a PD-1 inhibitor. In some embodiments, the PD-1 inhibitor is selected from PDR001 (Novartis), nivolumab (Bristol-Myers Squibb), pembrolizumab (Merck & Co.), pidilizumab (CureTech), MEDI0680 (Medimmune), REGN2810 (Regeneron), TSR-042 (Tesaro), PF-06801591 (Pfizer), BGB-A317 (Beigene), BGB-108 (Beigene), INCSHR1210 (Incyte), or AMP-224 (Amplimmune). In some embodiments, the PD-1 inhibitor is PDR001. PDR001 is also known as spartalizumab.

[0559] In certain embodiments, the combinations described herein include a LAG-3 inhibitor. In some embodiments, the LAG-3 inhibitor is selected from LAG525 (Novartis), BMS-986016 (Bristol-Myers Squibb), or TSR-033 (Tesaro).

[0560] In certain embodiments, the combinations described herein include a TIM-3 inhibitor. In some embodiments, the TIM-3 inhibitor is MBG453 (Novartis), TSR-022 (Tesaro), LY-3321367 (Eli Lilly), Sym23 (Symphogen), BGB-A425 (Beigene), INCAGN-2390 (Agenus), BMS-986258 (BMS), RO-7121661 (Roche), or LY-3415244 (Eli Lilly).

[0561] In certain embodiments, the combinations described herein include a PDL1 inhibitor. In one embodiment, the PDL1 inhibitor is selected from FAZ053 (Novartis), atezolizumab (Genentech), durvalumab (Astra Zeneca), or avelumab (Pfizer).

[0562] In certain embodiments, the combination described herein comprises GITR agonist.In some embodiments, the GITR agonist is selected from GWN323 (NVS), BMS-986156, MK-4166 or MK-1248 (Merck), TRX518 (Leap Therapeutics), INCAGN1876 (Incyte / Agenus), AMG 228 (Amgen) or INBRX-110 (Inhibrx).

[0563] In some embodiments, the combinations described herein include an IAP inhibitor. In some embodiments, the IAP inhibitor includes LCL161 or a compound disclosed in International Application No. PCT2008 / 016893.

[0564] In embodiments, the combination includes an mTOR inhibitor, for example, RAD001 (also known as everolimus).

[0565] In an embodiment, the combination includes an HDAC inhibitor, such as LBH589, which is also known as panobinostat.

[0566] In embodiments, the combination includes an IL-17 inhibitor, for example CJM112.

[0567] In certain embodiments, the combination described herein comprises an estrogen receptor (ER) antagonist.In some embodiments, the estrogen receptor antagonist is used in combination with a PD-1 inhibitor, a CDK4 / 6 inhibitor, or both.In some embodiments, the combination is used to treat ER-positive (ER+) cancer or breast cancer (for example, ER+ breast cancer).

[0568] In some embodiments, the estrogen receptor antagonist is a selective estrogen receptor degrader (SERD). SERD is an estrogen receptor antagonist, which binds to the receptor and, for example, causes receptor degradation or downregulation (Boer K. et al., (2017) Therapeutic Advances in Medical Oncology 9(7): 465-479). ER is a hormone-activated transcription factor that is important for the growth, development, and physiological function of the human reproductive system, for example. ER is activated by, for example, estrogen hormone (17 beta estradiol). ER expression and signal transduction are involved in cancer (e.g., breast cancer), for example, ER-positive (ER+) breast cancer. In some embodiments, the SERD is selected from LSZ102, fulvestrant, brilanestrant, or elacestrant.

[0569] In some embodiments, the SERD comprises a compound disclosed in International Application No. PCT2014 / 130310, which is incorporated herein by reference in its entirety.

[0570] In some embodiments, the SERD comprises LSZ102. LSZ102 has the chemical name: (E)-3-(4-((2-(2-(1,1-difluoroethyl)-4-fluorophenyl)-6-hydroxybenzo[b]thiophen-3-yl)oxy)phenyl)acrylic acid. In some embodiments, the SERD comprises fulvestrant (CAS Registry Number: 129453-61-8) or a compound disclosed in International Application PCT 2001 / 051056, which is incorporated by reference herein in its entirety. In some embodiments, the SERD comprises elacestrant (CAS Registry Number: 722533-56-4) or a compound disclosed in U.S. Pat. No. 7,612,114, which is incorporated by reference in its entirety. Elacestrant is also known as RAD1901, ER-306323, or (6R)-6-{2-[ethyl({4-[2-(ethylamino)ethyl]phenyl}methyl)amino]-4-methoxyphenyl}-5,6,7,8-tetrahydronaphthalen-2-ol. Elacestrant is an orally bioavailable non-steroidal combination of a selective estrogen receptor modulator (SERM) and a SERD. Elacestrant is also disclosed, for example, in Garner F et al., (2015) Anticancer Drugs 26(9):948-56. In some embodiments, the SERD is brillanestrat (CAS Registry Number: 1365888-06-7) or a compound disclosed in International Application PCT2015 / 136017, the entirety of which is incorporated by reference.

[0571] In some embodiments, the SERD is selected from RU 58668, GW7604, AZD9496, bazedoxifene, pipendoxifene, arzoxifene, OP-1074, or acolbifene, e.g., as disclosed in McDonell et al. (2015) Journal of Medicinal Chemistry 58(12) 4883-4887.

[0572] Other exemplary estrogen receptor antagonists are disclosed, for example, in WO 2011 / 156518, WO 2011 / 159769, WO 2012 / 037410, WO 2012 / 037411, and U.S. Patent No. 2012 / 0071535, all of which are incorporated herein by reference in their entireties.

[0573] In certain embodiments, the combinations described herein include a cyclin-dependent kinase 4 or 6 (CDK4 / 6) inhibitor. In some embodiments, the CDK4 / 6 inhibitor is used in combination with a PD-1 inhibitor, an estrogen receptor (ER) antagonist, or both. In some embodiments, the combination is used to treat ER-positive (ER+) cancer or breast cancer (e.g., ER+ breast cancer). In some embodiments, the CDK4 / 6 inhibitor is selected from ribociclib, abemaciclib (Eli Lilly), or palbociclib.

[0574] In some embodiments, the CDK4 / 6 inhibitor comprises ribociclib (CAS Registry Number: 1211441-98-3) or a compound disclosed in U.S. Pat. Nos. 8,415,355 and 8,685,980, which are incorporated by reference in their entireties.

[0575] In some embodiments, CDK4 / 6 inhibitors include compounds disclosed in International Application No. PCT2010 / 020675 and U.S. Patent Nos. 8,415,355 and 8,685,980, which are incorporated by reference in their entireties.

[0576] In some embodiments, the CDK4 / 6 inhibitor comprises ribociclib (CAS Registry Number: 1211441-98-3), which is also known as LEE011, KISQALI®, or 7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide.

[0577] In some embodiments, the CDK4 / 6 inhibitor comprises abemaciclib (CAS Registry Number: 1231929-97-7). Abemaciclib is also known as LY835219 or N-[5-[(4-ethyl-1-piperazinyl)methyl]-2-pyridinyl]-5-fluoro-4-[4-fluoro-2-methyl-1-(1-methylethyl)-1H-benzimidazol-6-yl]-2-pyrimidinamine. Abemaciclib is a selective CDK inhibitor for CDK4 and CDK6, and is disclosed, for example, in Torres-Guzman R et al. (2017) Oncotarget 10.18632 / oncotarget.17778.

[0578] In some embodiments, the CDK4 / 6 inhibitor comprises palbociclib (CAS Registry Number: 571190-30-2). Palbociclib is also known as PD-0332991, IBRANCE®, or 6-acetyl-8-cyclopentyl-5-methyl-2-{[5-(1-piperazinyl)-2-pyridinyl]amino}pyrido[2,3-d]pyrimidin-7(8H)-one. Palbociclib inhibits CDK4 with an IC50 of 11 nM and CDK6 with an IC50 of 16 nM, as disclosed, for example, in Finn et al. (2009) Breast Cancer Research 11(5):R77.

[0579] In certain embodiments, the combinations described herein include a chemokine (C-X-C motif) receptor 2 (CXCR2) inhibitor. In some embodiments, the CXCR2 inhibitor is selected from 6-chloro-3-((3,4-dioxo-2-(pentan-3-ylamino)cyclobut-1-en-1-yl)amino)-2-hydroxy-N-methoxy-N-methylbenzenesulfonamide, danilixin, reparixin, or navarixin.

[0580] In some embodiments, the CSF-1 / 1R binding agent is selected from a macrophage colony-stimulating factor (M-CSF) inhibitor, e.g., a monoclonal antibody or Fab to M-CSF (e.g., MCS110), a CSF-1R tyrosine kinase inhibitor (e.g., 4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide or BLZ945), a receptor tyrosine kinase inhibitor (RTK) (e.g., pexidartinib), or a CSF-1R targeting antibody (e.g., emactuzumab or FPA008). In some embodiments, the CSF-1 / 1R inhibitor is BLZ945. In some embodiments, the CSF-1 / 1R binding agent is MCS110. In other embodiments, the CSF-1 / 1R binding agent is pexidartinib.

[0581] In certain embodiments, the combination described herein includes a c-MET inhibitor. c-MET is a receptor tyrosine kinase that is overexpressed or mutated in many tumor cell types and plays a key role in tumor cell proliferation, survival, invasion, metastasis, and tumor angiogenesis. Inhibition of c-MET can induce cell death in tumor cells that overexpress c-MET protein or express constitutively activated c-MET protein. In some embodiments, the c-MET inhibitor is selected from capmatinib (INC280), JNJ-3887605, AMG337, LY2801653, MSC2156119J, crizotinib, tivantinib, or golvatinib.

[0582] In certain embodiments, the combinations described herein include a transforming growth factor beta (TGF-β, also known as TGFβ, TGFb, or TGF-beta, and used interchangeably herein) inhibitor. In some embodiments, the TGF-β inhibitor is selected from fresolimumab or XOMA089.

[0583] In certain embodiments, the combinations described herein include an adenosine A2a receptor (A2aR) antagonist (e.g., an A2aR pathway inhibitor, e.g., an adenosine inhibitor, e.g., an A2aR or CD-73 inhibitor). In some embodiments, the A2aR antagonist is used in combination with a PD-1 inhibitor and one or more (e.g., two, three, four, five, or all) of a CXCR2 inhibitor, a CSF-1 / 1R binder, a LAG-3 inhibitor, a GITR agonist, a c-MET inhibitor, or an IDO inhibitor. In some embodiments, the combination is used to treat pancreatic cancer, colorectal cancer, gastric cancer, or melanoma (e.g., refractory melanoma). In some embodiments, the A2aR antagonist is selected from PBF509 (NIR178) (Palobiofarma / Novartis), CPI444 / V81444 (Corvus / Genentech), AZD4635 / HTL-1071 (AstraZeneca / Heptares), bipadenant (Redox / Juno), GBV-2034 (Globavir), AB928 (Arcus Biosciences), theophylline, istradefylline (Kyowa Hakko Kogyo), tozadenant / SYN-115 (Acorda), KW-6356 (Kyowa Hakko Kogyo), ST-4206 (Leadiant Biosciences), or preladenant / SCH 420814 (Merck / Schering). Without wishing to be bound by theory, it is believed that in some embodiments, inhibition of A2aR leads to upregulation of IL-1b.

[0584] In certain embodiments, the combinations described herein include an inhibitor of indoleamine 2,3-dioxygenase (IDO) and / or tryptophan 2,3-dioxygenase (TDO). In some embodiments, the IDO inhibitor is used in combination with a PD-1 inhibitor and one or more (e.g., two, three, four, or all) of a TGF-β inhibitor, an A2aR antagonist, a CSF-1 / 1R binder, a c-MET inhibitor, or a GITR agonist. In some embodiments, the combination is used to treat pancreatic cancer, colorectal cancer, gastric cancer, or melanoma (e.g., refractory melanoma). In some embodiments, the IDO inhibitor is selected from (4E)-4-[(3-chloro-4-fluoroanilino)-nitrosomethylidene]-1,2,5-oxadiazol-3-amine (also known as epacadostat or INCB24360), indoximod (NLG8189), (1-methyl-D-tryptophan), α-cyclohexyl-5H-imidazo[5,1-a]isoindole-5-ethanol (also known as NLG919), indoximod, BMS-986205 (formerly F001287).

[0585] In certain embodiments, the combinations described herein include an inhibitor of a galectin, e.g., galectin-1 or galectin-3. In some embodiments, the combination includes a galectin-1 inhibitor and a galectin-3 inhibitor. In some embodiments, the combination includes a dual-specific inhibitor (e.g., a bispecific antibody molecule) that targets both galectin-1 and galectin-3. In some embodiments, the galectin inhibitor is used in combination with one or more therapeutic agents described herein. In some embodiments, the galectin inhibitor is selected from an anti-galectin antibody molecule, GR-MD-02 (Galectin Therapeutics), Galectin-3C (Mandal Med), Anginex, or OTX-008 (OncoEthix, Merck).

[0586] In some embodiments, the combination described herein comprises a MEK inhibitor.In some embodiments, the MEK inhibitor is selected from trametinib, selumetinib, AS703026, BIX02189, BIX02188, CI-1040, PD0325901, PD98059, U0126, XL-518, G-38963 or G02443714.In some embodiments, the MEK inhibitor is trametinib.

[0587] In one embodiment, the combination described herein includes an interleukin-1 beta (IL-1β) inhibitor. In some embodiments, the IL-1β inhibitor is selected from canakinumab, gevokizumab, anakinra, or rilonacept.

[0588] In certain embodiments, the combinations described herein include an IL-15 / IL-15Ra complex. In some embodiments, the IL-15 / IL-15Ra complex is selected from NIZ985 (Novartis), ATL-803 (Altor), or CYP0150 (Cytune).

[0589] In certain embodiments, the combination described herein comprises mouse double minute 2 homologue (MDM2) inhibitor.The human homologue of MDM2 is also known as HDM2.In some embodiments, the MDM2 inhibitor described herein is also known as HDM2 inhibitor.In some embodiments, the MDM2 inhibitor is selected from HDM201 or CGM097.

[0590] In embodiments, the MDM2 inhibitor comprises (S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one (also known as CGM097) or a compound disclosed in WO 2011 / 076786 for treating a disorder, e.g., a disorder described herein. In one embodiment, a therapeutic agent disclosed herein is used in combination with CGM097.

[0591] In some embodiments, the combinations described herein include a methylation inhibitor (HMA). In some embodiments, the HMA is selected from decitabine or azacitidine.

[0592] In certain embodiments, the combinations described herein include an inhibitor that acts on a pro-survival protein of the Bcl2 family. In certain embodiments, the combinations described herein include a Bcl-2 inhibitor. In some embodiments, the Bcl-2 inhibitor is venetoclax.

[0593] [ka] is.

[0594] In one embodiment, the Bcl-2 inhibitor is selected from compounds described in WO 2013 / 110890 and WO 2015 / 011400. In some embodiments, the Bcl-2 inhibitor comprises navitoclax (ABT-263), ABT-737, BP1002, SPC2996, APG-1252, obatoclax mesylate (GX15-070MS), PNT2258, Zn-d5, BGB-11417, or oblimersen (G3139). In some embodiments, the Bcl-2 inhibitor is (S)-5-(5-chloro-2-(3-(morpholinomethyl)-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)phenyl)-N-(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)-N-(4-hydroxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamide), Compound A1:

[0595] [ka] In some embodiments, the Bcl-2 inhibitor is N-(4-hydroxyphenyl)-3-[6-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide, compound A2:

[0596] [ka] is.

[0597] In one embodiment, the antibody-drug conjugate or combination disclosed herein is suitable for in vivo cancer treatment. For example, the combination may be used to inhibit the growth of cancerous tumors. The combination may also be used in combination with one or more of standard care and treatment (e.g., for cancer or infectious disorders), vaccines (e.g., therapeutic cancer vaccines), cell therapy, radiation therapy, surgery, or any other therapeutic agent or modality for treating the disorders herein. For example, the combination may be administered together with the antigen of interest to achieve antigen-specific enhancement of immunity. The combinations disclosed herein may be administered either sequentially or simultaneously.

[0598] Additional Embodiments The present disclosure provides the following additional embodiments to linker-drug groups, antibody-drug conjugates, linker groups, and methods of conjugation.

[0599] Linker-Drug Group In some embodiments, the linker-drug group of the present invention has the formula (A'):

[0600] [ka] (In the formula, R 1 is a reactive group, L1 is a bridging spacer, Lp is a divalent peptide spacer, G-L2-A is a self-immolative spacer, R 2 is the hydrophilic moiety, L2 is a bond, methylene, neopentylene, or C2-C3 alkenylene; A is a bond, -OC(=O)- * ,

[0601] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * , or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, L3 is a spacer moiety, D is a drug moiety that is capable of inhibiting the activity of MCl-1 protein when released from, for example, an antibody drug conjugate or immunoconjugate disclosed herein. or a pharmaceutically acceptable salt thereof.

[0602] Certain specific aspects and examples of linker-drug groups of the present invention are set forth in the list of enumerated embodiments below, it being recognized that the features specified in each embodiment can be combined with other specific features to provide further embodiments of the present invention.

[0603] Embodiment 1. R 1 is the reactive group, L1 is a bridging spacer, Lp is a bivalent peptide spacer containing 2 to 4 amino acid residues; G-L2-A is a self-immolative spacer, R 2 is the hydrophilic portion, L2 is a bond, methylene, neopentylene, or C2-C3 alkenylene; A is a bond, -OC(=O)- * ,

[0604] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- *, or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, L3 is the spacer portion, D is a drug moiety as defined herein, e.g., an MCl-1 inhibitor; A compound of formula (A'), or a pharmaceutically acceptable salt thereof:

[0605] Embodiment 2. R 1 is the reactive group, L1 is a bridging spacer, Lp is a bivalent peptide spacer containing 2 to 4 amino acid residues;

[0606] [ka] The base is

[0607] [ka] is selected from, where:

[0608] [ka] of * indicates the point of attachment to D (e.g., to the N or O of the drug moiety);

[0609] [ka] of *** indicates the point of attachment to Lp, R 2 is the hydrophilic portion, L2 is a bond, methylene, neopentylene, or C2-C3 alkenylene; A is a bond, -OC(=O)- * ,

[0610] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * , or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- * where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, L3 is the spacer portion, D is a drug moiety as defined herein, e.g., an MCl-1 inhibitor; A compound of formula (A'), or a pharmaceutically acceptable salt thereof:

[0611] Embodiment 3. Formula (B'):

[0612] [ka] (In the formula, R 1 is a reactive group, L1 is a bridging spacer, Lp is a bivalent peptide spacer containing 2 to 4 amino acid residues; R 2 is the hydrophilic moiety, A is a bond, -OC(=O)- * ,

[0613] [ka] , -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)- * , or -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)- *where each R a are independently selected from H, C1-C6 alkyl, and C3-C8 cycloalkyl; * indicates the point of attachment to D, L3 is a spacer moiety, D is a drug moiety, as defined herein, containing N or O, where D is connected to A via a direct bond from A to N or O of the drug moiety. A compound of formula (A') having the structure: or a pharmaceutically acceptable salt thereof.

[0614] Embodiment 4. R 1 but,

[0615] [ka] , -ONH2, -NH2,

[0616] [ka] , -N3,

[0617] [ka] , -SH, -SR 3 , -SSR 4 , -S(=O)2(CH=CH2), -(CH2)2S(=O)2(CH=CH2), -NHS(=O)2(CH=CH2), -NHC(=O)CH2Br, -NHC(=O)CH2I,

[0618] [ka] , -C(O)NHNH2,

[0619] [ka]

[0620] [ka] and L1 * -C(=O)(CH2) m O(CH2) m - ** ; * -C(=O)((CH2) m Or) t (CH2) n - ** ; * -C(=O)(CH2) m - ** ; * -C(=O)NH((CH2) m Or) t (CH2) n - ** ; * -C(=O)O(CH2) m SSC(R) 3 )2(CH2) m C(=O)NR 3 (CH2) m NR 3 C(=O)(CH2) m - ...

Claims

1. Formula (1): A-(L-D) ｐ (1) [In the formula, Ab is an anti-CD74 antibody or an antigen-binding fragment thereof; p is an integer from 1 to 16; -(L-D) is a compound of the formula (C): 【Chemistry 1】 (In the formula, R １ is a linking group, L １ is a bridging spacer, L ｐ is a peptide group containing 1 to 6 amino acids, D is an Mcl-1 inhibitor, the Mcl-1 inhibitor having the formula (II): 【Chemistry 2】 (In the formula, Z 0 is a nitrogen atom or a C—R 04 group, R 01 is a halogen atom, a linear or branched (C 1 -C 6 ) alkyl group, a linear or branched (C 2 -C 6 ) alkenyl group, a linear or branched (C 2 -C 6 ) alkynyl group, a linear or branched (C 1 -C 6 ) haloalkyl group, a hydroxy group, a linear or branched (C 1 -C 6 ) alkoxy group, a -S-(C 1 -C 6 ) alkyl group, a cyano group, -Cy 08 , -NR 011 R 011 '; R 02 , R 03 and R 04 each independently represent a hydrogen atom, a halogen atom, a linear or branched (C 1 -C 6 ) alkyl group, a linear or branched (C 2 -C 6 ) alkenyl group, a linear or branched (C 2 -C 6 ) alkynyl group, a linear or branched (C 1 -C 6 ) haloalkyl, a hydroxy group, a linear or branched (C 1 -C 6 ) alkoxy group, -S-(C 1 -C 6 ) alkyl group, a cyano group, a nitro group, -(C 0 -C 6 ) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C 0 -C 6 ) alkyl-Cy 01 , -(C 2 -C 6 ) alkenyl-Cy 01 , -(C 2 -C 6 )alkynyl-Cy 01 , -O-(C 1 -C 6 )alkyl-NR 011 R 011 ', -O-(C 1 -C 6 )alkyl-R 031 , -C(O)-OR 011 , -O-C(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C 1 -C 6 )alkyl-NR 011 -C(O)-R 011 ', -SO 2 -NR 011 R 011 ', or -SO 2 -(C 1 -C 6 )alkyl; or the pair (R 02 , R 03 ) or (R 03 , R 04 ) together with the carbon atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, said ring being optionally substituted with a group selected from linear or branched (C 1 -C 6 ) alkyl, -NR 013 R 013 ', -(C 0 -C 6 ) alkyl-Cy 01 and oxo; R 06 and R 07 are each independently a hydrogen atom, a halogen atom, a linear or branched (C 1 -C 6 ) alkyl group, a linear or branched (C 2 -C 6 ) alkenyl group, a linear or branched (C 2 -C 6 ) alkynyl group, a linear or branched (C 1 -C 6 ) haloalkyl, a hydroxy group, a linear or branched (C 1 -C 6 ) alkoxy group, -S-(C 1 -C 6 ) alkyl group, a cyano group, a nitro group, -(C 0 -C 6 ) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C 0 -C 6 ) alkyl-Cy 01 , -(C 2 -C 6 ) alkenyl-Cy 01 , -(C 2 -C 6 ) alkynyl-Cy 01 , -O-(C 1 -C 6 )alkyl-R 012 , -C(O)-OR 011 , -O-C(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C 1 -C 6 )alkyl-NR 011 -C(O)-R 011 ', -SO 2 -NR 011 R 011 ', or -SO 2 -(C 1 -C 6 )alkyl; or the pair (R 06 , R 07 ), when fused with two adjacent carbon atoms, together with the carbon atom to which they are attached, form an aromatic or non-aromatic ring containing from 5 to 7 ring members, optionally containing from 1 to 3 heteroatoms selected from O, S and N, said resulting ring being optionally substituted with groups selected from linear or branched (C 1 -C 6 )alkyl groups, -NR 013 R 013 ', -(C 0 -C 6 )alkyl-Cy 01 and oxo; R 08 is a hydrogen atom, a linear or branched (C 1 -C 8 ) alkyl group, an aryl group, a heteroaryl group, an aryl-(C 1 -C 6 ) alkyl group or a heteroaryl(C 1 -C 6 ) alkyl group; R 09 is a linear or branched (C 1 -C 6 ) alkyl group, a linear or branched (C 2 -C 6 ) alkenyl group, a linear or branched (C 2 -C 6 ) alkynyl group, -Cy 02 , -(C 1 -C 6 ) alkyl-Cy 02 , -(C 2 -C 6 ) alkenyl-Cy 02 , -(C 2 -C 6 ) alkynyl-Cy 02 , -Cy 02 -Cy 03 , -(C 2 -C 6 ) alkynyl-O-Cy 02 , -Cy 02 -(C 0 -C 6 ) alkyl-O-(C 0 -C 6 ) alkyl-Cy 03 , a halogen atom, a cyano group, -C(O)-R 014 , -C(O)-NR 014 R 014 ', R 011 and R 011 ′ are each independently a hydrogen atom, an optionally substituted linear or branched (C 1 -C 6 ) alkyl group, or —(C 0 -C 6 ) alkyl-Cy 01 ; or the pair (R 011 , R 011 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to said nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein said N atom is optionally substituted with a linear or branched (C 1 -C 6 ) alkyl group, wherein one or more of the carbon atoms of said linear or branched (C 1 -C 6 ) alkyl group are optionally deuterated; R 012 is -Cy 05 , -Cy 05 -(C 0 -C 6 )alkyl-Cy 06 , -Cy 05 -(C 0 -C 6 )alkyl-O-(C 0 -C 6 )alkyl-Cy 06 , -Cy 05 -(C 0 -C 6 )alkyl-NR 011 -(C 0 -C 6 )alkyl-Cy 06 , -Cy 05 -Cy 06 -O-(C 0 -C 6 )alkyl-Cy 07 , -Cy 05 -(C 0 -C 6 )alkyl-Cy 09 , -NH-C(O)-NH-R 011 , -C(O)-NR 011 R 011 ', -NR 011 R 011 ', -OR 011 , -NR 011 -C(O)-R 011 ', -O-(C 1 -C 6 )alkyl-OR 011 , -SO 2 -R 011 , or -C(O)-OR 011 ; R 013 , R 013 ', R 014 and R 014 ' are each independently a hydrogen atom or an optionally substituted linear or branched (C 1 -C 6 ) alkyl group; Cy 01 , Cy 02 , Cy 03 , Cy 05 , Cy 06 , Cy 07 and Cy 08 are each independently a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group, each of which is optionally substituted; Cy09 is 【Chemistry 3】 and R 015 is a hydrogen atom; a -(CH 2 ) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 group; a linear or branched (C 1 -C 6 ) alkoxy(C 1 -C 6 ) alkyl group; a -U 0 -(CH 2 ) q0 -NR 021 R 021 ' group; or a -(CH 2 ) r0 -U 0 -(CH 2 ) s0 -heterocycloalkyl group; R 016 is a hydrogen atom; a hydroxy group; a hydroxy(C 1 -C 6 )alkyl group; -(CH 2 ) r0 -U 0 -(CH 2 ) s0 -heterocycloalkyl group; a (CH 2 ) r0 -U 0 -V 0 -O-P(O)(OR 020 ) 2 group; a -O-P(O)(O - M + ) 2 group; a -O-S(O) 2 OR 020 group; a -S(O) 2 OR 020 group; a -(CH 2 ) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 group; a -(CH 2 ) p0 -O-C(O)-NR 022 R 023 group; or -U 0 -(CH 2 ) q0 -NR 021 R 021 ' group; R 017 is a hydrogen atom; a -(CH 2 ) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 group; a -CH 2 -P(O)(OR 020 ) 2 group, a -O-P(O)(OR 020 ) 2 group; a -O-P(O)(O - M + ) 2 group; a hydroxy group; a hydroxy(C 1 -C 6 )alkyl group; a -(CH 2 ) r0 -U 0 -(CH 2 ) s0 -heterocycloalkyl group; a -U 0 -(CH 2 ) q0 -NR 021 R 021 ' group; or an aldonic acid; M + is a pharma- ceutically acceptable monovalent cation; U 0 is a bond or an oxygen atom; V 0 is a —(CH 2 ) s0 — group or a —C(O)— group; R 018 is a hydrogen atom or a (C 1 -C 6 )alkoxy(C 1 -C 6 )alkyl group; R 019 is a hydrogen atom or a hydroxy(C 1 -C 6 )alkyl group; R 020 is a hydrogen atom or a linear or branched (C 1 -C 6 ) alkyl group; R 021 and R 021 ′ are each independently a hydrogen atom, a linear or branched (C 1 -C 6 ) alkyl group, or a hydroxy(C 1 -C 6 ) alkyl group; or the pair (R 021 , R 021 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to said nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein said resulting ring is optionally substituted with hydrogen atoms or linear or branched (C 1 -C 6 ) alkyl groups; R 022 is a (C 1 -C 6 )alkoxy(C 1 -C 6 )alkyl group, a -(CH 2 ) p0 -NR 024 R 024 ' group or a -(CH 2 ) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 20 group; R 023 is a hydrogen atom or a (C 1 -C 6 )alkoxy(C 1 -C 6 )alkyl group; or the pair (R 022 , R 023 ) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 18 ring members, which optionally contains, in addition to said nitrogen atom, 1 to 5 heteroatoms selected from O, S and N, wherein said resulting ring is optionally substituted with hydrogen atoms, linear or branched (C 1 -C 6 ) alkyl groups or heterocycloalkyl groups; R 024 and R 024 ′ are each independently a hydrogen atom or a linear or branched (C 1 -C 6 ) alkyl group; or the pair (R 024 , R 024 ′) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring consisting of 5 to 7 ring members, which may contain, in addition to said nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein said resulting ring is optionally substituted with hydrogen atoms or linear or branched (C 1 -C 6 ) alkyl groups; R 027 is a hydrogen atom or a linear or branched (C 1 -C 6 ) alkyl group; R 028 represents a -O-P(O)(O - )(O - ) group, a -O-P(O)(O - )(OR 030 ) group, a -O-P(O)(OR 030 )(OR 030 ') group, a -(CH 2 ) p0 -O-SO 2 -O - group, a -(CH 2 ) p0 -SO 2 -O - group, a -(CH 2 ) p0 -O-SO 2 -OR 030 group, -Cy 010 , a -(CH 2 ) p0 -SO 2 -OR 030 group, a -O-C(O)-R 029 group, a -O-C(O)-OR 029 group or a -O-C(O)-NR 029 R 029 ' is a base, R 029 and R 029 ′ are each independently a hydrogen atom, a linear or branched (C 1 -C 6 ) alkyl group or a linear or branched amino(C 1 -C 6 ) alkyl group; R 030 and R 030 ′ are each independently a hydrogen atom, a linear or branched (C 1 -C 6 ) alkyl group or an aryl (C 1 -C 6 ) alkyl group; R031 is 【Chemistry 4】 and p 0 is an integer equal to 0, 1, 2 or 3; q 0 is an integer equal to 1, 2, 3, or 4; r 0 and s 0 are independently integers equal to 0 or 1; one of R 03 , R 09 and R 012 is covalently attached to said bridging spacer; or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts of any of the foregoing, G １ -L ２ -A is a self-immolative spacer, L ２ is a bond, methylene, neopentylene or C ２ ~C ３ alkenylene, A is a bond, -OC(=O)- ＊ , 【Chemistry 5】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, L ３ is a spacer moiety, R ２ is the hydrophilic portion) [The following is a list of Antibody-drug conjugates of.

2. -(LD) is a compound of the formula (D): 【Chemistry 6】 (In the formula, R １ is a linking group, L １ is a bridging spacer, Lp is a peptide group containing 1 to 6 amino acids; A is a bond, -OC(=O)- ＊ , 【Chemistry 7】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, L ３ is a spacer moiety, R ２ is the hydrophilic portion) 2. The antibody-drug conjugate of claim 1, or a pharma- ceutically acceptable salt thereof.

3. (1) L １ but, 【Chemistry 8】 or ＊ -CH(OH)CH(OH)CH(OH)CH(OH)- ＊＊ Including, where each n is an integer from 1 to 12, where L １ of ＊ indicates a direct or indirect attachment point to Lp, and L １ of ＊＊ is R １ indicating the point of direct or indirect attachment to (2) L １ but, 【Chemistry 9】 and n is an integer from 1 to 12, or n is 1, or n is 12, where L １ of ＊ indicates a direct or indirect attachment point to Lp, and L １ of ＊＊ is R １ indicating the point of direct or indirect attachment to (3) L １ but, 【Chemistry 10】 and n is an integer from 1 to 12, where L １ of ＊ indicates a direct or indirect attachment point to Lp, and L １ of ＊＊ is R １ indicating the point of direct or indirect attachment to (4) L １ but, 【Chemistry 11】 Wherein L １ of ＊ indicates a direct or indirect attachment point to Lp, and L １ of ＊＊ is R １ indicating a direct or indirect point of attachment to (5) L １ but, ＊ -C(=O)(CH ２ ) ｍ O(CH ２ ) ｍ - ＊＊ ; ＊ -C(=O)((CH ２ ) ｍ O) ｔ (CH ２ ) ｎ - ＊＊ ; ＊ -C(=O)(CH ２ ) ｍ - ＊＊ ; ＊ -C(=O)NH((CH ２ ) ｍ O) ｔ (CH ２ ) ｎ - ＊＊ ; ＊ -C(=O)O(CH ２ ) ｍ SSC(R ３ ) ２ (CH ２ ) ｍ C(=O)NR ３ (CH ２ ) ｍ NR ３ C(=O)(CH ２ ) ｍ - ＊＊ ; ＊ -C(=O)O(CH ２ ) ｍ C(=O)NH(CH ２ ) ｍ - ＊＊ ; ＊ -C(=O)(CH ２ ) ｍ NH(CH ２ ) ｍ - ＊＊ ; ＊ -C(=O)(CH ２ ) ｍ NH(CH ２ ) ｎ C(=O)- ＊＊ ; ＊ -C(=O)(CH ２ ) ｍ X １ (CH ２ ) ｍ - ＊＊ ; ＊ -C(=O)((CH ２ ) ｍ O) ｔ (CH ２ ) ｎ X １ (CH ２ ) ｎ - ＊＊ ; ＊ -C(=O)(CH ２ ) ｍ NHC(=O)(CH ２ ) ｎ - ＊＊ ; ＊ -C(=O)((CH ２ ) ｍ O) ｔ (CH ２ ) ｎ NHC(=O)(CH ２ ) ｎ - ＊＊ ; ＊ -C(=O)(CH ２ ) ｍ NHC(=O)(CH ２ ) ｎ X １ (CH ２ ) ｎ - ＊＊ ; ＊ -C(=O)((CH ２ ) ｍ O) ｔ (CH ２ ) ｎ NHC(=O)(CH ２ ) ｎ X １ (CH ２ ) ｎ - ＊＊ ; ＊ -C(=O)((CH ２ ) ｍ O) ｔ (CH ２ ) ｎ C(=O)NH(CH ２ ) ｍ - ＊＊ ; ＊ -C(=O)(CH ２ ) ｍ C(R ３ ) ２ - ＊＊ または ＊ -C(=O)(CH ２ ) ｍ C(=O)NH(CH ２ ) ｍ - ＊＊ where L is a bridging spacer comprising １ of ＊ indicates a direct or indirect attachment point to Lp, and L １ of ＊＊ is R １ indicates the point of direct or indirect attachment to X １ teeth, 【Chemistry 12】 and each m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each n is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; each t is independently selected from 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, and 30; Each R ３ is H and C １ ~C ６ independently selected from alkyl, The antibody-drug conjugate according to claim 1 or 2.

4. (1) R ２ is a polyethylene glycol, a polyalkylene glycol, a polyol, a polysarcosine, a sugar, an oligosaccharide, a polypeptide, or 1 to 3 【Chemistry 13】 C substituted with a group ２ ~C ６ is an alkyl-containing hydrophilic moiety, (2) R ２ but, 【Chemistry 14】 where n is an integer between 1 and 6. 【Chemistry 15】 That is, (3) The hydrophilic portion is (i) Polysarcosine, for example, having the moiety: 【Chemistry 16】 (wherein n is an integer between 3 and 25, and R is H, -CH ３ or -CH ２ CH ２ C(═O)OH), or (ii) Formula: 【Chemistry 17】 Polyethylene glycol: (Wherein, R is H, —CH ３ , C.H. ２ CH ２ NHC(=O)OR ａ , -CH ２ CH ２ NHC(=O)R ａ or -CH ２ CH ２ C(=O)OR ａ and R' is OH, -OCH ３ , -CH ２ CH ２ NHC(=O)OR ａ , -CH ２ CH ２ NHC(=O)R ａ or -OCH ２ CH ２ C(=O)OR ａ where R ａ is H, OH or C １～４ C optionally substituted with any one of alkoxyl １～４ alkyl, and each of m and n is independently an integer between 2 and 25. or (4) The hydrophilic portion is 【Chemistry 18】 Including, The antibody-drug conjugate according to any one of claims 1 to 3.

5. (i) L ３ But the structure 【Chemistry 19】 (In the formula, Wは、-CH ２ -、-CH ２ O-、-CH ２ N(R ｂ )C(=O)O-、-NHC(=O)C(R ｂ ) ２ NHC(=O)O-、 -NHC(=O)C(R ｂ ) ２ HUNTER ｂ ) ２ HUNTER ２ N(X-R ２ )C(=O)O-、-C(=O)N(X-R ２ )-、-CH ２ N(X-R ２ )C(=O)-、-C(=O)NR ｂ -、-C(=O)NH-、-CH ２ NR ｂ C(=O)-、-CH ２ NR ｂ C(=O)NH-,-CH ２ NR ｂ P.S. ｂ -、-NHC(=O)-, -NHC(=O)NH-, -OC(=O)NH-、 -S (O) ２ NH-, -NHS(O) ２ -, -C(=O)-, -C(=O)O-, or -NH-, where each R ｂ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; X is a bond, triazolyl or -CH ２ -triazolyl- or (ii) L ３ But the structure 【Chemistry 20】 (In the formula, Wは、-CH ２ -、-CH ２ O-、-CH ２ N(R ｂ )C(=O)O-、-NHC(=O)C(R ｂ ) ２ NHC(=O)O-、 -NHC(=O)C(R ｂ ) ２ NH-、-NHC(=O)C(R ｂ ) ２ NHC(=O)-、-CH ２ N(X-R ２ )C(=O)O-、-C(=O)N(X-R ２ )-、 -CH ２ N(X-R ２ )C(=O)-、-C(=O)NR ｂ -、-C(=O)NH-、-CH ２ NR ｂ C(=O)-、-CH ２ NR ｂ C(=O)NH-、 -CH ２ NR ｂ P.S. ｂ -、-NHC(=O)-, -NHC(=O)NH-, -OC(=O)NH-、 -S (O) ２ NH-, -NHS(O) ２ -, -C(=O)-, -C(=O)O-, or -NH-, where each R ｂ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; X is -CH ２ -Triazolyl-C １～４ Alkylene-OC(O)NHS(O) ２ NH-, -C ４～６ Cycloalkylene-OC(O)NHS(O) ２ NH-, -(CH ２ CH ２ O) ｎ -C(O)NHS(O) ２ NH-, -(CH ２ CH ２ O) ｎ -C(O)NHS(O) ２ NH-(CH ２ CH ２ O) ｎ - or -CH ２ -Triazolyl-C １～４ Alkylene-OC(O)NHS(O) ２ NH-(CH ２ CH ２ O) ｎ -, where each n is independently 1, 2, or 3. The antibody-drug conjugate according to any one of claims 1 to 4, wherein the spacer moiety has the formula:

6. (1) the linking group is formed by a reaction involving at least one reactive group; (2) The bonding group is a first reactive group attached to the bridging spacer; a second reactive group that is attached to the antibody or antigen-binding fragment or is an amino acid residue of the antibody or antigen-binding fragment; is formed by reacting (3) At least one of the reactive groups is Thiol, Maleimide, Haloacetamides, Azide, Alkynes, Cyclooctene, Triarylphosphines, oxanorbornadiene, Cyclooctyne, Diaryltetrazines, Monoaryltetrazines, Norbornene, aldehyde, Hydroxylamine, Hydrazine, NH ２ -NH-C(=O)-、 Ketones, vinyl sulfone, Aziridine, Amino acid residues, 【Chemistry 21】 、-ONH ２ 、-NH ２ 、 【Chemical 22】 、-N ３ 、 【Chemistry 23】 、-SH、-SR ３ 、-SSR ４ 、-S(=O) ２ (CH=CH ２ )、-(CH ２ ) ２ S(=O) ２ (CH=CH ２ )、-NHS(=O) ２ (CH=CH ２ )、-NHC(=O)CH ２ Br、-NHC(=O)CH ２ I、 【Chemistry 24】 (O)NHNH ２ 、 【Chemistry 25】 Including, Where: Each R ３ is H and C １ ~C ６ independently selected from alkyl, Each R ４ is 2-pyridyl or 4-pyridyl, Each R ５ is H, C １ ~C ６ independently selected from alkyl, F, Cl and -OH; Each R ６ is H, C １ ~C ６ Alkyl, F, Cl, -NH ２ , -OCH ３ , -OCH ２ CH ３ , -N(CH ３ ) ２ , -CN, -NO ２ and -OH, Each R ７ is H, C １～６ Alkyl, fluoro, benzyloxy substituted with -C(=O)OH, benzyl substituted with -C(=O)OH, C substituted with -C(=O)OH １～４ Alkoxy and -C(=O)OH substituted C １～４ independently selected from alkyl, (4) The first reactive group and the second reactive group are Thiols and maleimides, Thiols and haloacetamides, Thiols and vinyl sulfones, Thiols and aziridines, Azides and alkynes, Azide and cyclooctyne, Azide and cyclooctene, Azides and triarylphosphines, Azides and oxanorbornadienes, diaryltetrazines and cyclooctenes, Monoaryltetrazines and norbornenes, Aldehydes and hydroxylamines, Aldehydes and hydrazines, Aldehydes and NH ２ -NH-C(=O)-, Ketones and hydroxylamines, Ketones and hydrazines, Ketones and NH ２ -NH-C(=O)-, Hydroxylamine and 【Chemistry 26】 、 Amines and 【Chemistry 27】 ,or CoA and serine residues or (5) The bonding group is 【Chemistry 28】 【change】 【change】 【change】 and Where: R ３２ is H, C １～４ alkyl, phenyl, pyrimidine or pyridine; R ３５ is H, C １～６ Alkyl, phenyl, or C substituted with 1 to 3 -OH groups １～４ is alkyl, Each R ７ is H, C １～６ Alkyl, fluoro, benzyloxy substituted with -C(=O)OH, benzyl substituted with -C(=O)OH, C substituted with -C(=O)OH １～４ Alkoxy and -C(=O)OH substituted C １～４ independently selected from alkyl, R ３７ is independently selected from H, phenyl and pyridine; q is 0, 1, 2 or 3; R ８ is H or methyl, R ９ is H, -CH ３ or phenyl.

7. (1) the peptide group represented by Lp comprises 1 to 4 amino acid residues, 1 to 3 amino acid residues, or 1 or 2 amino acid residues, optionally selected from L-glycine (Gly), L-valine (Val), L-citrulline (Cit), L-cysteic acid (sulfo-Ala), L-lysine (Lys), L-isoleucine (Ile), L-phenylalanine (Phe), L-methionine (Met), L-asparagine (Asn), L-proline (Pro), L-alanine (Ala), L-leucine (Leu), L-tryptophan (Trp) and L-tyrosine (Tyr); (2) the peptide group comprises Val-Cit, Phe-Lys, Val-Ala, Val-Lys, Leu-Cit, Sulfo-Ala-Val and / or Sulfo-Ala-Val-Ala, or (3) Lp is 【Chemical Formula 29】 The antibody-drug conjugate according to any one of claims 1 to 6, selected from:

8. -(L-D) is a compound of the formula: （１） 【Chemistry 30】 (In the formula, R is H, -CH ３ or -CH ２ CH ２ C(=O)OH, A is a bond, -OC(=O)- ＊ , 【Chemistry 31】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （２） 【Chemistry 32】 (In the formula, R is H, -CH ３ or -CH ２ CH ２ C(=O)OH, A is a bond, -OC(=O)- ＊ , 【Chemical 33】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （３） 【Chemical 34】 (In the formula, R is H, -CH ３ or -CH ２ CH ２ C(=O)OH, A is a bond, -OC(=O)- ＊ , 【Chemistry 35】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （４） 【Chemical 36】 (In the formula, Each R is H, —CH ３ and -CH ２ CH ２ C(═O)OH; A is a bond, -OC(=O)- ＊ , 【Chemical 37】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （５） 【Chemical Formula 38】 (In the formula, Each R is H, —CH ３ and -CH ２ CH ２ C(═O)OH; A is a bond, -OC(=O)- ＊ , 【Chemical 39】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （６） 【Chemistry 40】 (In the formula, Xa is -CH ２ --, --OCH ２ --, --NHCH ２ -or- NRCH ２ -, and each R is independently H, -CH ３ or -CH ２ CH ２ C(=O)OH, A is a bond, -OC(=O)- ＊ , 【Chemistry 41】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （７） 【Chemistry 42】 (In the formula, R is H, -CH ３ or -CH ２ CH ２ C(=O)OH, A is a bond, -OC(=O)- ＊ , 【Chemistry 43】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （８） 【Chemistry 44】 (In the formula, Xb is -CH ２ --, --OCH ２ --NHCH ２ -or- NRCH ２ -, and each R is independently H, -CH ３ or -CH ２ CH ２ C(=O)OH, A is a bond, -OC(=O)- ＊ , 【Chemistry 45】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （９） 【Chemistry 46】 (In the formula, A is a bond, -OC(=O)- ＊ , 【Chemistry 47】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （１０） 【Chemistry 48】 (In the formula, A is a bond, -OC(=O)- ＊ , 【Chemistry 49】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （１１） 【Chemistry 50】 (In the formula, A is a bond, -OC(=O)- ＊ , 【Chemistry 51】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （１２） 【Chemistry 52】 (In the formula, A is a bond, -OC(=O)- ＊ , 【Chemistry 53】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （１３） 【Chemical 54】 (In the formula, A is a bond, -OC(=O)- ＊ , 【Chemistry 55】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor. （１４） 【Chemistry 56】 (In the formula, A is a bond, -OC(=O)- ＊ , 【Chemistry 57】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor, or （１５） 【Chemistry 58】 (In the formula, Each R is independently H, -CH ３ or -CH ２ CH ２ C(=O)OH, A is a bond, -OC(=O)- ＊ , 【Chemistry 59】 , -OC(=O)N(CH ３ ) CH ２ CH ２ N (CH ３ ) C(=O)- ＊ Or -OC(=O)N(CH ３ ) C(R ａ ) ２ C(R ａ ) ２ N (CH ３ ) C(=O)- ＊ and Here, each R ａ is H, C １ ~C ６ Alkyl and C ３ ~C ８ cycloalkyl; ＊ indicates the point of attachment to D, D is an Mcl-1 inhibitor.

8. The antibody-drug conjugate of claim 1 , comprising or formed from a compound of the formula:

9. A is a bond and / or R is -CH ３ The antibody-drug conjugate of claim 8,

10. D is represented by formula (III): 【Chemistry 60】 (In the formula, R ０１ is linear or branched (C １ ~C ６ ) alkyl group, R ０３ is -O-(C １ ~C ６ ) alkyl-NR ０１１ R ０１１ 'or 【Chemistry 61】 and Here, R ０１１ and R ０１１ ' are each independently a hydrogen atom, an optionally substituted linear or branched (C １ ~C ６ ) alkyl group, or -(C ０ ~C ６ ) Alkyl-Cy ０１ and Or against (R ０１１ , R ０１１ ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to said nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, where said N atom is a hydrogen atom or a linear or branched (C １ ~C ６ ) alkyl groups, Here, R ０２７ is a hydrogen atom, R ０２８ is -(CH ２ ) ｐ０ -O-SO ２ -O － group or -(CH ２ ) ｐ０ -SO ２ -OR ０３０ It is based on p 0 is an integer equal to 0, 1, 2 or 3; R ０９ is linear or branched (C ２ ~C ６ ) an alkynyl group or -Cy ０２ and R ０１２ is -Cy ０５ , -Cy ０５ - (C ０ ~C ６ ) Alkyl-Cy ０６ Or -Cy ０５ - (C ０ ~C ６ ) Alkyl-Cy ０９ and Cy ０１ , Cy ０２ , Cy ０５ and Cy ０６ are, independently of each other, a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group, each of which is optionally substituted; Cy ０９ teeth 【Chemistry 62】 and R ０１５ , R ０１６ and R ０１７ is as defined for formula (II), wherein one of R03, R09 or R012 is covalently attached to said bridging spacer. or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts of any of the foregoing.

11. (1) R ０１ is methyl or ethyl; (2) R ０３ Ga-O-CH ２ -CH ２ -NR ０１１ R ０１１ '(in the formula, R ０１１ and R ０１１ ', together with the nitrogen atom which holds them, are hydrogen atoms or linear or branched (C １ ~C ６ ) an alkyl group) which forms a piperazinyl group, which may be substituted with a group; (3) R ０３ But the formula: 【Chemistry 63】 (In the formula, R ０２７ is a hydrogen atom, R ０２８ Ha-(CH ２ ) ｐ０ -SO ２ -OR ０３０ (Based on Including, (4) R ０３ But the formula: 【Chemistry 64】 (In the formula, 【Chemistry 65】 is a bond to the bridging spacer) Including, (5) R ０９ Cy ０２ That is, (6) Cy ０５ comprises a heteroaryl group selected from a pyrazolyl group and a pyrimidinyl group; (7) Cy ０５ is a pyrimidinyl group, and / or (8) The L is a group represented by the formula (II) or (III) ０３ or said L is linked to D by a covalent bond from L to R of formula (II) or (III) ０９ is attached to D by a covalent bond The antibody-drug conjugate of claim 10.

12. (1) D is, 【Chemistry 66】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (2) D: 【Chemistry 67】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (3) D is 【Chemistry 68】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (4) D is 【Chemistry 69】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (5) D is 【Chemistry 70】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (6) D is 【Chemistry 71】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (7) D is 【Chemical Formula 72】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (8) D is 【Chemical 73】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (9) D is 【Chemical Formula 74】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (10) D is 【Chemistry 75】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (11) D is 【Chemical 76】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (12) D is 【Chemical Formula 77】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof, or (13) D is 【Chemical 78】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts thereof; (14) D is 【Chemical 79】 or enantiomers, diastereoisomers, atropisomers, deuterated derivatives, and / or pharma- ceutically acceptable salts of any of the foregoing, or (15)-(LD) is formed from a compound selected from Table A or an enantiomer, diastereoisomer, atropisomer, deuterated derivative, and / or a pharma- ceutically acceptable salt thereof; The antibody-drug conjugate according to any one of claims 1 to 11.

13. The antibody-drug conjugate of claim 1, wherein -(LD) comprises or is formed from the following compound: 【Chemistry 80】 【change】 【change】

14. the anti-CD74 antibody or antigen-binding fragment thereof (1) three heavy chain CDRs and three light chain CDRs as follows: (i) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 1, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 16, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 70, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18; (ii) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 4, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 16, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 70, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18; (iii) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:19, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:20, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:21; (iv) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 8, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 22, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 20, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18; (v) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:1, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:35, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:70, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:18; (vi) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 4, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 35, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 70, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18; (vii) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:71, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:20, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:21; or (viii) a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 8, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 17, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 20, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18; (2) (I) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 23; (II) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 27; (III) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 31; (IV) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 36; (V) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 40; or (VI) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 10, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 44; or (3) (a) a heavy chain having the amino acid sequence of SEQ ID NO: 12, and a light chain having the amino acid sequence of SEQ ID NO: 25; (b) a heavy chain having the amino acid sequence of SEQ ID NO: 14, and a light chain having the amino acid sequence of SEQ ID NO: 25; (c) a heavy chain having the amino acid sequence of SEQ ID NO: 15, and a light chain having the amino acid sequence of SEQ ID NO: 25; (d) a heavy chain having the amino acid sequence of SEQ ID NO: 12, and a light chain having the amino acid sequence of SEQ ID NO: 29; (e) a heavy chain having the amino acid sequence of SEQ ID NO: 14, and a light chain having the amino acid sequence of SEQ ID NO: 29; (f) a heavy chain having the amino acid sequence of SEQ ID NO: 15, and a light chain having the amino acid sequence of SEQ ID NO: 29; (g) a heavy chain having the amino acid sequence of SEQ ID NO: 12, and a light chain having the amino acid sequence of SEQ ID NO: 33; (h) a heavy chain having the amino acid sequence of SEQ ID NO: 14, and a light chain having the amino acid sequence of SEQ ID NO: 33; (i) a heavy chain having the amino acid sequence of SEQ ID NO: 15, and a light chain having the amino acid sequence of SEQ ID NO: 33; (j) a heavy chain having the amino acid sequence of SEQ ID NO: 12, and a light chain having the amino acid sequence of SEQ ID NO: 38; (k) a heavy chain having the amino acid sequence of SEQ ID NO: 14, and a light chain having the amino acid sequence of SEQ ID NO: 38; (l) a heavy chain having the amino acid sequence of SEQ ID NO: 15, and a light chain having the amino acid sequence of SEQ ID NO: 38; (m) a heavy chain having the amino acid sequence of SEQ ID NO: 12, and a light chain having the amino acid sequence of SEQ ID NO: 42; (n) a heavy chain having the amino acid sequence of SEQ ID NO: 14, and a light chain having the amino acid sequence of SEQ ID NO: 42; (o) a heavy chain having the amino acid sequence of SEQ ID NO: 15, and a light chain having the amino acid sequence of SEQ ID NO: 42; (p) a heavy chain having the amino acid sequence of SEQ ID NO: 12, and a light chain having the amino acid sequence of SEQ ID NO: 46; (q) a heavy chain having the amino acid sequence of SEQ ID NO: 14, and a light chain having the amino acid sequence of SEQ ID NO: 46; or (r) the antibody drug conjugate of any one of claims 1 to 13, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 15, and a light chain having the amino acid sequence of SEQ ID NO:

46.

15. 15. A composition comprising multiple copies of the antibody-drug conjugate of any one of claims 1 to 14, wherein the average p of the antibody-drug conjugates in the composition is 2 to 16, 2 to 8, or 2 to 4.

16. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 1 to 14 or the composition of claim 15, and a pharma- ceutically acceptable carrier.

17. A pharmaceutical composition for use in a method for treating a subject having or suspected of having cancer, comprising a therapeutically effective amount of an antibody-drug conjugate according to any one of claims 14, a composition according to claim 15, or a pharmaceutical composition according to claim 16, (1) the cancer expresses CD74; (2) The cancer is a tumor or blood cancer. (3) the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies derived from T cells or B cells, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer or splenic cancer, and / or (4) The method further comprises administering to the subject at least one additional therapeutic agent. The pharmaceutical composition.

18. A pharmaceutical composition for use in a method of reducing or inhibiting the growth of a tumor in a subject or a method of reducing or slowing the expansion of a cancer cell population in a subject, comprising a therapeutically effective amount of an antibody-drug conjugate according to any one of claims 1 to 14, a composition according to claim 15, or a pharmaceutical composition according to claim 16, (1) the tumor or the cancer cell population expresses CD74; (2) the tumor is breast cancer, gastric cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, hepatocellular carcinoma, melanoma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer or splenic cancer, or the cancer cell population is derived from a tumor or blood cancer; (3) The cancer cell population is derived from breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies derived from T cells or B cells, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer, or splenic cancer. (4) administration of the antibody-drug conjugate, composition or pharmaceutical composition reduces or inhibits the growth of the tumor or reduces or slows the expansion of the cancer cell population by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%; and / or (5) The method further comprises administering to the subject at least one additional therapeutic agent. The pharmaceutical composition.

19. The additional therapeutic agent is navitoclax, ABT-737, BP1002, SPC2996, APG-1252, obatoclax mesylate, PNT2258, Zn-d5, BGB-11417, oblimersen, venetoclax, (S)-5-(5-chloro-2-(3-(morpholinomethyl)-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)phenyl)-N-(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)-N-(4-hydroxyphenyl)-2-[(2-methylphenyl)-1H-pyrrol-3-yl] ...

19. The pharmaceutical composition of claim 17 or 18, wherein the Bcl-2 inhibitor is selected from N-(4-hydroxyphenyl)-3-[6-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide.

20. 17. A pharmaceutical composition for use in a method for determining whether a subject having or suspected of having cancer will respond to treatment with an antibody-drug conjugate of any one of claims 1 to 14, a composition of claim 15, or a pharmaceutical composition of claim 16, said method comprising providing a biological sample from said subject; contacting said sample with said antibody-drug conjugate; and detecting binding of said antibody-drug conjugate to cancer cells in said sample, (1) the cancer cells in the sample express CD74; (2) the cancer expresses CD74; (3) The cancer is a tumor or a blood cancer. (4) the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, gastric cancer, acute myeloid leukemia, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia, follicular lymphoma, lymphoid malignancies derived from T cells or B cells, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, chronic lymphocytic leukemia, prostate cancer, small cell lung cancer or splenic cancer, and / or (5) The sample is a tissue biopsy sample, a blood sample, or a bone marrow sample. The pharmaceutical composition.

21. A method of making an antibody-drug conjugate according to any one of claims 1 to 14, comprising reacting an antibody or antigen-binding fragment with a cleavable linker linked to an MCL1 inhibitor under conditions allowing conjugation.

22. Formula (1): A-(L-D) ｐ (1) (In the formula, Ab is an anti-CD74 antibody or an antigen-binding fragment thereof, wherein said Ab is an Fc-silenced antibody; p is an integer from 1 to 16; L is a linker, D is an MCL1 inhibitor compound, said Mcl-1 inhibitor having the formula (II): 【Chemistry 81】 (In the formula, Z 0 is a nitrogen atom or a C—R 04 group, R 01 is a halogen atom, a linear or branched (C 1 -C 6 ) alkyl group, a linear or branched (C 2 -C 6 ) alkenyl group, a linear or branched (C 2 -C 6 ) alkynyl group, a linear or branched (C 1 -C 6 ) haloalkyl group, a hydroxy group, a linear or branched (C 1 -C 6 ) alkoxy group, a -S-(C 1 -C 6 ) alkyl group, a cyano group, -Cy 08 , -NR 011 R 011 '; R 02 , R 03 and R 04 each independently represent a hydrogen atom, a halogen atom, a linear or branched (C 1 -C 6 ) alkyl group, a linear or branched (C 2 -C 6 ) alkenyl group, a linear or branched (C 2 -C 6 ) alkynyl group, a linear or branched (C 1 -C 6 ) haloalkyl, a hydroxy group, a linear or branched (C 1 -C 6 ) alkoxy group, -S-(C 1 -C 6 ) alkyl group, a cyano group, a nitro group, -(C 0 -C 6 ) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C 0 -C 6 ) alkyl-Cy 01 , -(C 2 -C 6 ) alkenyl-Cy 01 , -(C 2 -C 6 )alkynyl-Cy 01 , -O-(C 1 -C 6 )alkyl-NR 011 R 011 ', -O-(C 1 -C 6 )alkyl-R 031 , -C(O)-OR 011 , -O-C(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C 1 -C 6 )alkyl-NR 011 -C(O)-R 011 ', -SO 2 -NR 011 R 011 ', or -SO 2 -(C 1 -C 6 )alkyl; or the pair (R 02 , R 03 ) or (R 03 , R 04 ) together with the carbon atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, optionally containing 1 to 3 heteroatoms selected from O, S and N, said ring being optionally substituted with a group selected from linear or branched (C 1 -C 6 ) alkyl, -NR 013 R 013 ', -(C 0 -C 6 ) alkyl-Cy 01 and oxo; R 06 and R 07 are each independently a hydrogen atom, a halogen atom, a linear or branched (C 1 -C 6 ) alkyl group, a linear or branched (C 2 -C 6 ) alkenyl group, a linear or branched (C 2 -C 6 ) alkynyl group, a linear or branched (C 1 -C 6 ) haloalkyl, a hydroxy group, a linear or branched (C 1 -C 6 ) alkoxy group, -S-(C 1 -C 6 ) alkyl group, a cyano group, a nitro group, -(C 0 -C 6 ) alkyl-NR 011 R 011 ', -O-Cy 01 , -(C 0 -C 6 ) alkyl-Cy 01 , -(C 2 -C 6 ) alkenyl-Cy 01 , -(C 2 -C 6 ) alkynyl-Cy 01 , -O-(C 1 -C 6 )alkyl-R 012 , -C(O)-OR 011 , -O-C(O)-R 011 , -C(O)-NR 011 R 011 ', -NR 011 -C(O)-R 011 ', -NR 011 -C(O)-OR 011 ', -(C 1 -C 6 )alkyl-NR 011 -C(O)-R 011 ', -SO 2 -NR 011 R 011 ', or -SO 2 -(C 1 -C 6 )alkyl; or the pair (R 06 , R 07 ), when fused with two adjacent carbon atoms, together with the carbon atom to which they are attached, form an aromatic or non-aromatic ring containing from 5 to 7 ring members, optionally containing from 1 to 3 heteroatoms selected from O, S and N, said resulting ring being optionally substituted with groups selected from linear or branched (C 1 -C 6 )alkyl groups, -NR 013 R 013 ', -(C 0 -C 6 )alkyl-Cy 01 and oxo; R 08 is a hydrogen atom, a linear or branched (C 1 -C 8 ) alkyl group, an aryl group, a heteroaryl group, an aryl-(C 1 -C 6 ) alkyl group or a heteroaryl(C 1 -C 6 ) alkyl group; R 09 is a linear or branched (C 1 -C 6 ) alkyl group, a linear or branched (C 2 -C 6 ) alkenyl group, a linear or branched (C 2 -C 6 ) alkynyl group, -Cy 02 , -(C 1 -C 6 ) alkyl-Cy 02 , -(C 2 -C 6 ) alkenyl-Cy 02 , -(C 2 -C 6 ) alkynyl-Cy 02 , -Cy 02 -Cy 03 , -(C 2 -C 6 ) alkynyl-O-Cy 02 , -Cy 02 -(C 0 -C 6 ) alkyl-O-(C 0 -C 6 ) alkyl-Cy 03 , a halogen atom, a cyano group, -C(O)-R 014 , -C(O)-NR 014 R 014 ', R 011 and R 011 ′ are each independently a hydrogen atom, an optionally substituted linear or branched (C 1 -C 6 ) alkyl group, or —(C 0 -C 6 ) alkyl-Cy 01 ; or the pair (R 011 , R 011 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to said nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein said N atom is optionally substituted with a linear or branched (C 1 -C 6 ) alkyl group, wherein one or more of the carbon atoms of said linear or branched (C 1 -C 6 ) alkyl group are optionally deuterated; R 012 is -Cy 05 , -Cy 05 -(C 0 -C 6 )alkyl-Cy 06 , -Cy 05 -(C 0 -C 6 )alkyl-O-(C 0 -C 6 )alkyl-Cy 06 , -Cy 05 -(C 0 -C 6 )alkyl-NR 011 -(C 0 -C 6 )alkyl-Cy 06 , -Cy 05 -Cy 06 -O-(C 0 -C 6 )alkyl-Cy 07 , -Cy 05 -(C 0 -C 6 )alkyl-Cy 09 , -NH-C(O)-NH-R 011 , -C(O)-NR 011 R 011 ', -NR 011 R 011 ', -OR 011 , -NR 011 -C(O)-R 011 ', -O-(C 1 -C 6 )alkyl-OR 011 , -SO 2 -R 011 , or -C(O)-OR 011 ; R 013 , R 013 ', R 014 and R 014 ' are each independently a hydrogen atom or an optionally substituted linear or branched (C 1 -C 6 ) alkyl group; Cy 01 , Cy 02 , Cy 03 , Cy 05 , Cy 06 , Cy 07 and Cy 08 are each independently a cycloalkyl group, a heterocycloalkyl group, an aryl group, or a heteroaryl group, each of which is optionally substituted; Cy09 is 【Chemistry 82】 and R 015 is a hydrogen atom; a -(CH 2 ) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 group; a linear or branched (C 1 -C 6 ) alkoxy(C 1 -C 6 ) alkyl group; a -U 0 -(CH 2 ) q0 -NR 021 R 021 ' group; or a -(CH 2 ) r0 -U 0 -(CH 2 ) s0 -heterocycloalkyl group; R 016 is a hydrogen atom; a hydroxy group; a hydroxy(C 1 -C 6 )alkyl group; -(CH 2 ) r0 -U 0 -(CH 2 ) s0 -heterocycloalkyl group; a (CH 2 ) r0 -U 0 -V 0 -O-P(O)(OR 020 ) 2 group; a -O-P(O)(O - M + ) 2 group; a -O-S(O) 2 OR 020 group; a -S(O) 2 OR 020 group; a -(CH 2 ) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 group; a -(CH 2 ) p0 -O-C(O)-NR 022 R 023 group; or -U 0 -(CH 2 ) q0 -NR 021 R 021 ' group; R 017 is a hydrogen atom; a -(CH 2 ) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 020 group; a -CH 2 -P(O)(OR 020 ) 2 group, a -O-P(O)(OR 020 ) 2 group; a -O-P(O)(O - M + ) 2 group; a hydroxy group; a hydroxy(C 1 -C 6 )alkyl group; a -(CH 2 ) r0 -U 0 -(CH 2 ) s0 -heterocycloalkyl group; a -U 0 -(CH 2 ) q0 -NR 021 R 021 ' group; or an aldonic acid; M + is a pharma- ceutically acceptable monovalent cation; U 0 is a bond or an oxygen atom; V 0 is a —(CH 2 ) s0 — group or a —C(O)— group; R 018 is a hydrogen atom or a (C 1 -C 6 )alkoxy(C 1 -C 6 )alkyl group; R 019 is a hydrogen atom or a hydroxy(C 1 -C 6 )alkyl group; R 020 is a hydrogen atom or a linear or branched (C 1 -C 6 ) alkyl group; R 021 and R 021 ′ are each independently a hydrogen atom, a linear or branched (C 1 -C 6 ) alkyl group, or a hydroxy(C 1 -C 6 ) alkyl group; or the pair (R 021 , R 021 ') together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 7 ring members, which optionally contains, in addition to said nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein said resulting ring is optionally substituted with hydrogen atoms or linear or branched (C 1 -C 6 ) alkyl groups; R 022 is a (C 1 -C 6 )alkoxy(C 1 -C 6 )alkyl group, a -(CH 2 ) p0 -NR 024 R 024 ' group or a -(CH 2 ) p0 -O-(CHR 018 -CHR 019 -O) q0 -R 20 group; R 023 is a hydrogen atom or a (C 1 -C 6 )alkoxy(C 1 -C 6 )alkyl group; or the pair (R 022 , R 023 ) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring containing 5 to 18 ring members, which optionally contains, in addition to said nitrogen atom, 1 to 5 heteroatoms selected from O, S and N, wherein said resulting ring is optionally substituted with hydrogen atoms, linear or branched (C 1 -C 6 ) alkyl groups or heterocycloalkyl groups; R 024 and R 024 ′ are each independently a hydrogen atom or a linear or branched (C 1 -C 6 ) alkyl group; or the pair (R 024 , R 024 ′) together with the nitrogen atom to which they are attached form an aromatic or non-aromatic ring consisting of 5 to 7 ring members, which may contain, in addition to said nitrogen atom, 1 to 3 heteroatoms selected from O, S and N, wherein said resulting ring is optionally substituted with hydrogen atoms or linear or branched (C 1 -C 6 ) alkyl groups; R 027 is a hydrogen atom or a linear or branched (C 1 -C 6 ) alkyl group; R 028 represents a -O-P(O)(O - )(O - ) group, a -O-P(O)(O - )(OR 030 ) group, a -O-P(O)(OR 030 )(OR 030 ') group, a -(CH 2 ) p0 -O-SO 2 -O - group, a -(CH 2 ) p0 -SO 2 -O - group, a -(CH 2 ) p0 -O-SO 2 -OR 030 group, -Cy 010 , a -(CH 2 ) p0 -SO 2 -OR 030 group, a -O-C(O)-R 029 group, a -O-C(O)-OR 029 group or a -O-C(O)-NR 029 R 029 ' is a base, R 029 and R 029 ′ are each independently a hydrogen atom, a linear or branched (C 1 -C 6 ) alkyl group or a linear or branched amino(C 1 -C 6 ) alkyl group; R 030 and R 030 ′ are each independently a hydrogen atom, a linear or branched (C 1 -C 6 ) alkyl group or an aryl (C 1 -C 6 ) alkyl group; R031 is 【Chemistry 83】 and p 0 is an integer equal to 0, 1, 2 or 3; q 0 is an integer equal to 1, 2, 3, or 4; r 0 and s 0 are independently integers equal to 0 or 1; one of R 03 , R 09 and R 012 is covalently bonded to said linker; or an enantiomer, diastereoisomer, atropisomer, deuterated derivative, and / or a pharma- ceutically acceptable salt of any of these. Antibody-drug conjugates of.

23. 1. A pharmaceutical composition for use in a method for treating a disease or disorder, comprising navitoclax, ABT-737, BP1002, SPC2996, APG-1252, obatoclax mesylate, PNT2258, Zn-d5, BGB-11417, oblimersen, venetoclax, (S)-5-(5-chloro-2-(3-(morpholinomethyl)-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)phenyl)-N-(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)-N-(4-hydroxyphenyl 23. The pharmaceutical composition comprising the antibody-drug conjugate of claim 22 in combination with a Bcl-2 inhibitor selected from N-(4-hydroxyphenyl)-3-[6-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide, ...2-dimethyl-1H-pyrrole-3-carboxamide, N-(4-hydroxyphenyl)-3-[6-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide, N-(4-hydroxyphenyl)-3-[6-[(

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