EPHA2 BCL-XL inhibitor antibody-drug conjugates and methods of use thereof

Abstract

An anti-EphA2 antibody drug conjugate that binds to a human oncology target is disclosed. The antibody drug conjugates comprise a Bcl-xL inhibitor drug moiety and an anti-EphA2 antibody or antigen-binding fragment thereof that binds to an antigen target, such as an antigen expressed on a tumor or other cancer cell. The disclosure further relates to methods and compositions for treating cancer by administering the antibody drug conjugates provided herein. Also disclosed are linker drug conjugates comprising the Bcl-xL inhibitor drug moieties and methods of making the same.

Description

[0001] Related applications

[0002] This application claims the benefit under 35 U.S.C. §119(e) of the filing date of U.S. Provisional Application No. 63 / 344,454, filed on May 20, 2022, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present disclosure relates to antibody-drug conjugates (ADCs) comprising a Bcl-xL inhibitor and an anti-EphA2 antibody or antigen-binding fragment thereof that binds to an antigenic target (e.g., an antigen expressed on a tumor or other cancer cell). The present disclosure further relates to methods and compositions for treating and / or diagnosing cancers that express the target antigen and / or are amenable to treatment by modulating Bcl-xL expression and / or activity, as well as methods for preparing such compositions. Also disclosed are linker-drug conjugates comprising a Bcl-xL inhibitor drug moiety and methods for preparing the same. Background Art

[0004] Apoptosis (programmed cell death) is an evolutionarily conserved pathway that is essential for tissue homeostasis, development, and the removal of damaged cells. Dysregulation of apoptosis can lead to human diseases, including malignancies, neurodegenerative diseases, immune system diseases, and autoimmune diseases (Hanahan and Weinberg, Cell. 2011 Mar 4; 144(5): 646-74; Marsden and Strasser, Annu Rev Immunol. 2003; 21: 71-105; Vaux and Flavell, Curr Opin Immunol. 2000 Dec; 12(6): 719-24). Escape of apoptosis is considered a hallmark of cancer, involved in the development and continued proliferation of tumors and resistance to anticancer treatments (Hanahan and Weinberg, Cell. 2000 Jan 7; 100(1): 57-70).

[0005] The Bcl-2 family of proteins comprises key regulators of cell survival that can inhibit (e.g., Bcl-2, Bcl-xL, Mcl-1) or promote (e.g., Bad, Bax) apoptosis (Gross et al., Genes Dev. 1999 Aug 1;13(15):1899-911, Youle and Strasser, Nat. Rev. Mol. Cell Biol. 2008 Jan;9(1):47-59).

[0006] In response to stress, whether a cell survives or undergoes apoptosis depends on the degree of pairing between Bcl-2 family members that promote cell death and those that promote cell survival. In most cases, these interactions involve the docking of the Bcl-2 homology 3 (BH3) domain of the pro-apoptotic family member into a groove on the surface of the pro-survival member. The presence of Bcl-2 homology (BH) domains defines the members of the Bcl-2 family, which are divided into three main groups based on the specific BH domains present in the protein. Pro-survival members such as Bcl-2, Bcl-xL, and Mcl-1 contain BH domains 1-4, while pro-apoptotic effectors Bax and Bak, which permeabilize the mitochondrial outer membrane during apoptosis, contain BH domains 1-3 (Youle and Strasser, Nat. Rev. Mol. Cell Biol. 2008 Jan;9(1):47-59).

[0007] Overexpression of pro-survival members of the Bcl-2 family is a hallmark of cancer, and these proteins have been shown to play an important role in tumor development, maintenance, and resistance to anticancer therapies (Czabotar et al., Nat. Rev. Mol. Cell Biol. 2014 Jan;15(1):49-63). Bcl-xL (also known as BCL2L1, derived from BCL2-like 1) is frequently amplified in cancer (Beroukhim et al., Nature 2010 Feb 18;463(7283):899-905), and its expression has been shown to be inversely correlated with sensitivity to more than 120 anticancer therapeutic molecules in a representative panel of cancer cell lines (NCI-60) (Amundson et al., Cancer Res. 2000 Nov 1;60(21):6101-10).

[0008] In addition, several studies using transgenic knockout mouse models and transgenic overexpression of Bcl-2 family members have highlighted the importance of these proteins in immune system diseases and autoimmune diseases (for review, see Merino et al., Apoptosis 2009 Apr;14(4):570-83. doi:10.1007 / s10495-008-0308-4. PMID:19172396). Transgenic overexpression of Bcl-xL in the T cell compartment resulted in resistance to apoptosis induced by glucocorticoids, g-irradiation, and CD3 cross-linking, suggesting that transgenic Bcl-xL overexpression can reduce apoptosis in resting and activated T cells (Droin et al., Biochim Biophys Acta 2004 Mar 1;1644(2-3):179-88. doi:10.1016 / j.bbamcr.2003.10.011. PMID:14996502). In patient samples, persistent or high expression of anti-apoptotic Bcl-2 family proteins was observed (Pope et al., Nat Rev Immunol. 2002 Jul; 2(7): 527-35. doi: 10.1038 / nri846. PMID: 12094227). In particular, T cells isolated from the joints of patients with rheumatoid arthritis showed increased expression of Bcl-xL and were able to resist spontaneous apoptosis (Salmon et al., J Clin Invest. 1997 Feb 1; 99(3): 439-46. doi: 10.1172 / JCI119178. PMID: 9022077).

[0009] These findings have led to the discovery and development of a new class of drugs called BH3 mimetics. These molecules are able to disrupt the interaction between pro-apoptotic and anti-apoptotic members of the Bcl-2 family and are potent inducers of apoptosis. These new drugs include inhibitors of Bcl-2, Bcl-xL, Bcl-w, and Mcl-1. The first BH3 mimetics described were ABT-737 and ABT-263, which target Bcl-2, Bcl-xL, and Bcl-w (Park et al., J. Med. Chem. 2008 Nov 13; 51(21): 6902-15; Roberts et al., J. Clin. Oncol. 2012 Feb 10; 30(5): 488-96). Subsequently, selective inhibitors of Bcl-2 (ABT-199 and S55746) were discovered – Souers et al., Nat Med. 2013 Feb;19(2):202-8; Casara et al., Oncotarget 2018 Apr 13;9(28):20075-20088), Bcl-xL (A-1155463 and A-1331852 – Tai et al., ACS Med Chem Lett. 2014 Aug 26;5(10):1088-93; Leverson et al., Sci Transl Med. 2015 Mar 18;7(279):279ra40), and Mcl-1 (A-1210477, S63845, S64315, AMG-176, and AZD-5991 – Leverson et al., Cell Death Res. 2016 Mar;18(2):279ra40). Dis. 2015 Jan 15;6:e1590.; Kotschy et al., Nature 2016, 538, 477-482; Maragno et al., AACR 2019, Poster #4482; Kotschy et al., WO 2015 / 097123; Caenepeel et al., Cancer Discov. 2018 Dec;8(12):1582-1597; Tron et al., Nat. Commun. 2018 Dec 17;9(1):5341). The selective Bcl-2 inhibitor ABT-199 is now approved for use in combination therapy for patients with CLL and AML, while other inhibitors are still in preclinical or clinical development.In preclinical models, ABT-263 has shown activity in a variety of hematological malignancies and solid tumors (Shoemaker et al., Clin. Cancer Res. 2008 Jan 1; 14(11): 3268-77; Ackler et al., Cancer Chemother. Pharmacol. 2010 Oct; 66(5): 869-80; Chen et al., Mol. Cancer Ther. 2011 Dec; 10(12): 2340-9). In clinical studies, ABT-263 has shown objective antitumor activity in lymphoid malignancies (Wilson et al., Lancet Oncol. 2010 Dec; 11(12): 1149-59; Roberts et al., J. Clin. Oncol. 2012 Feb 10; 30(5): 488-96), and its activity is being studied in combination with several solid tumor therapies. Selective Bcl-xL inhibitors A-1155463 or A-1331852 showed in vivo activity in preclinical models of T-ALL (T cell acute lymphoblastic leukemia) and different types of solid tumors (Tao et al., ACS Med. Chem. Lett. August 26, 2014; 5(10): 1088-93; Leverson et al., Sci. Transl. Med. March 18, 2015; 7(279): 279ra40). The use of BH3 mimetics has also shown benefits in preclinical models of immune system diseases and autoimmune diseases. Treatment with ABT-737 (Bcl-2, Bcl-xL and Bcl-w inhibitor) can effectively inhibit lymphocyte proliferation in vitro. Importantly, mice treated with ABT-737 showed a significant reduction in disease severity in animal models of arthritis and lupus (Bardwell et al., J Clin Invest. 1997 Feb 1; 99(3): 439-46. doi: 10.1172 / JCI119178. PMID: 9022077). In addition, studies have shown that ABT-737 prevents allogeneic T cell activation, proliferation, and cytotoxicity in vitro and inhibits allogeneic T cell and B cell responses after skin transplantation, with high selectivity for lymphocytes (Cippa et al., Transpl Int. 2011 Jul; 24(7): 722-32. doi: 10.1111 / j.1432-2277.2011.01272.x. Epub 2011 May 25. PMID: 21615547). Therefore, therapeutic targeting of Bcl-xL or its upstream and / or downstream proteins in the apoptosis signaling pathway represents an attractive approach for the development of new therapies in oncology as well as in the fields of immune and autoimmune diseases.

[0010] The EphA2 receptor belongs to the ephrin receptor subfamily of receptor tyrosine kinases. Studies have shown that EphA2 is produced in high quantities in tumor tissues, while present at relatively low levels in most normal adult tissues. EphA2 dysregulation is associated with various pathological processes, particularly cancer. In certain types of cancer, EphA2 is associated with poor prognosis and decreased patient survival. Therefore, the EphA2 receptor is an attractive target for antibody-drug conjugates. Summary of the Invention

[0011] In some embodiments, the present disclosure provides, in part, novel antibody-drug conjugate (ADC) compounds having biological activity against cancer cells. These compounds can slow, inhibit and / or reverse tumor growth in mammals, and / or can be used to treat human cancer patients. In some embodiments, the present disclosure is more specifically directed to ADC compounds that can bind to and kill cancer cells. In some embodiments, the ADC compounds disclosed herein comprise a linker that attaches a Bcl-xL inhibitor to a full-length anti-EphA2 antibody or antigen-binding fragment. In some embodiments, the ADC compound can also be internalized into the target cell after binding.

[0012] In some embodiments, the ADC compound can be represented by formula (1):

[0013] Ab-(LD) p (1)

[0014] wherein Ab is an anti-EphA2 antibody or an antigen-binding fragment thereof;

[0015] D is a Bcl-xL inhibitor;

[0016] L is a linker that covalently attaches Ab to D; and

[0017] p is an integer from 1 to 16. In some embodiments, Ab is an antibody or antigen-binding fragment thereof that targets cancer cells.

[0018] In some embodiments, for the ADC compound of formula (1), D comprises a Bcl-xL inhibitor compound of formula (I') or formula (II') covalently attached to a linker L:

[0019]

[0020] or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein:

[0021] R1 and R2 independently of one another represent a group selected from the group consisting of: hydrogen; a) a linear or branched C1-C6 alkyl group optionally substituted by a hydroxyl group or a C1-C6 alkoxy group; a) a C3-C6 cycloalkyl group; a trifluoromethyl group; and a linear or branched C1-C6 alkylene-heterocycloalkyl group, wherein the heterocycloalkyl group is optionally substituted by a linear or branched C1-C6 alkyl group;

[0022] or R1 and R2 together with the carbon atom carrying them form a C3-C6 cycloalkylene group,

[0023] R3 represents a group selected from the group consisting of: hydrogen; a) C3-C6 cycloalkyl; linear or branched C1-C6 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ;-X1-OR c ;-X1-COOR c ;-X1-PO(OH)2;-X1-SO2(OH);

[0024] -X1-N3 and:

[0025]

[0026] R a and R b represents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR d R e ; C1-C6 alkylene-N + R d R e R f ; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a C1-C6 alkoxy group; and the group:

[0027]

[0028] or R a and R b Together with the nitrogen atom that carries them, they form ring B1;

[0029] or R a 、Rb and R c C3-C8 heterocycloalkyl groups forming bridges with the nitrogen atoms carrying them,

[0030] R c 、R d 、R e 、R f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0031] or R d and R e Together with the nitrogen atoms that carry them, they form ring B2,

[0032] or R d 、R e and R f C3-C8 heterocycloalkyl groups forming bridges with the nitrogen atoms carrying them,

[0033] Het1 represents a group selected from the group consisting of:

[0034]

[0035] Het2 represents a group selected from the group consisting of:

[0036]

[0037] A1 is -NH-, -N(C1-C3 alkyl), O, S or Se,

[0038] A2 is N, CH or C(R5),

[0039] G is selected from the group consisting of:

[0040] -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , -C1-C6 alkyl optionally substituted by hydroxyl groups, halogen, -NO2 and -CN, wherein:

[0041] -R G1 and R G2 are independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C1-C6 alkyl substituted by hydroxy, C1-C6 alkyl substituted by a C1-C6 alkoxy group, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl;

[0042] -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; -R G1 and R G2 Together with the atoms to which they are attached, they form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of:

[0043]

[0044] where R G4 is selected from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C1-C6 alkyl substituted by hydroxy, C1-C6 alkyl substituted by a C1-C6 alkoxy group, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl,

[0045] and R G5 represents a hydrogen atom or a C1-C6 alkyl group optionally substituted by 1 to 3 halogen atoms,

[0046] R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group,

[0047] R5 represents a group selected from the group consisting of: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; C2-C6 alkenyl; C2-C6 alkynyl; halogen; and -CN,

[0048] R6 represents a group selected from the group consisting of:

[0049] hydrogen;

[0050] a linear or branched-C1-C6 alkylene-R8 group;

[0051] -C2-C6 alkenyl;

[0052] -X2-O-R7;

[0053]

[0054] -X2-NSO2-R7;

[0055] -C=C(R9)-Y1-O-R7;

[0056] C3-C6 cycloalkyl;

[0057] C3-C6 heterocycloalkyl optionally substituted by a hydroxy group;

[0058] C3-C6 cycloalkylene-Y2-R7;

[0059] C3-C6 heterocycloalkylene-Y2-R7 group, and

[0060] a heteroarylene-R7 group optionally substituted by a linear or branched C1-C6 alkyl group,

[0061] R7 represents a group selected from the group consisting of: a linear or branched C1-C6 alkyl group; (C3-C6) cycloalkylene-R8;

[0062]

[0063] wherein Cy represents a C3-C8 cycloalkyl group,

[0064] R8 represents a group selected from the group consisting of: hydrogen; a straight or branched C1-C6 alkyl group, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' aR' b R' c ;-O-R' c ;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b ;-X'2-NR' a R' b ;-NR' c -X'2-N3 and

[0065] R9 represents a group selected from the group consisting of a linear or branched C1-C6 alkyl group, a trifluoromethyl group, a hydroxyl group, a halogen group, and a C1-C6 alkoxy group,

[0066] R 10 represents a radical selected from the group consisting of hydrogen, fluorine, chlorine, bromine, -CF3 and methyl,

[0067] R 11 represents a group selected from the group consisting of hydrogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8 and C3-C8 heterocycloalkylene-CH2-R8,

[0068] R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group,

[0069] R 14 and R 15 independently of one another represent hydrogen or a methyl group, or R 14 and R 15 Together with the carbon atoms that carry them, they form cyclohexyl groups,

[0070] R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0071] X1 and X2 independently represent a linear or branched chain

[0072] C1-C6 alkylene group, which is optionally substituted by one or two groups selected from the group consisting of trifluoromethyl, hydroxy, halogen and C1-C6 alkoxy,

[0073] X'2 represents a linear or branched C1-C6 alkylene group,

[0074] R' a and R' b represents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ;

[0075] C1-C6 alkylene-N + R' d R' e R' f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a hydroxyl group or a C1-C6 alkoxy group; and the group:

[0076]

[0077] or R' a and R' b Together with the nitrogen atoms that carry them, they form ring B3,

[0078] or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them,

[0079] R' c , R' d , R' e , R' f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0080] or R' d and R' e Together with the nitrogen atoms that carry them, they form ring B4,

[0081] or R' d , R' e and R' f C3-C bridges with the nitrogen atoms that carry them 8D Heterocycloalkyl,

[0082] Y1 represents a linear or branched C1-C4 alkylene group,

[0083] Y2 represents bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-, -C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO- or -NH-SO2-,

[0084] m=0, 1 or 2,

[0085] B1, B2, B3 and B4 are independently C3-C8 heterocycloalkyl groups, which may be: (i) monocyclic or bicyclic, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) substituted by one or two groups selected from the group consisting of fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl,

[0086] -NH2, oxo and piperidinyl, wherein one of the R3 and, if present, R8 groups is covalently attached to the linker and wherein the valence of the atom is not exceeded by one or more substituents bonded thereto; or

[0087]

[0088] or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein:

[0089] n=0, 1 or 2,

[0090] ------ indicates a single bond or a double bond,

[0091] A4 and A5 independently represent a carbon or nitrogen atom,

[0092] Z1 represents a bond, -N(R)- or -O-, wherein R represents hydrogen or a linear or branched C1-C6 alkyl group,

[0093] R1 represents a group selected from the group consisting of: hydrogen; a) a linear or branched C1-C6 alkyl group optionally substituted by a hydroxyl group or a C1-C6 alkoxy group; a) a C3-C6 cycloalkyl group; a trifluoromethyl group; and a linear or branched C1-C6 alkylene-heterocycloalkyl group, wherein the heterocycloalkyl group is optionally substituted by a linear or branched C1-C6 alkyl group;

[0094] R2 represents hydrogen or methyl;

[0095] R3 represents a group selected from the group consisting of: hydrogen; a linear or branched C1-C4 alkyl group; -X1-NR a R b ;-X1-N + R a R b R c ;-X1-OR c ;-X1-COOR c ; -X1-PO(OH)2; -X1-SO2(OH); -X1-N3 and:

[0096]

[0097] R a and R b represents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR d R e ; C1-C6 alkylene-N + R d R e R f ; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a C1-C6 alkoxy group; and the group:

[0098]

[0099] or R a and R b Together with the nitrogen atom that carries them, they form ring B1;

[0100] or R a 、R b and R c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom that carries them,

[0101] R c 、R d 、R e 、R f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0102] or R d and R e Together with the nitrogen atoms that carry them, they form ring B2,

[0103] or R d 、R e and R f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them,

[0104] Het1 represents a group selected from the group consisting of:

[0105]

[0106] Het2 represents a group selected from the group consisting of:

[0107]

[0108] A1 is -NH-, -N(C1-C3 alkyl), O, S or Se,

[0109] A2 is N, CH or C(R5),

[0110] G is selected from the group consisting of:

[0111] -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NRG1 )NR G1 R G2 , -C1-C6 alkyl optionally substituted by hydroxyl groups, halogen, -NO2 and -CN, wherein:

[0112] -RG1 and RG2 are each independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C1-C6 alkyl substituted by hydroxy, C1-C6 alkyl substituted by a C1-C6 alkoxy group, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl;

[0113] -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; -R G1 and R G2 Together with the atoms to which they are attached, they form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of:

[0114]

[0115] where R G4 is selected from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C1-C6 alkyl substituted by hydroxy, C1-C6 alkyl substituted by a C1-C6 alkoxy group, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl,

[0116] and R G5 represents a hydrogen atom or a C1-C6 alkyl group optionally substituted by 1 to 3 halogen atoms,

[0117] R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl group, a hydroxyl group or a methoxy group,

[0118] R5 represents a group selected from the group consisting of: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; C2-C6 alkenyl; C2-C6 alkynyl; halogen; and -CN,

[0119] R6 represents a group selected from the group consisting of:

[0120] hydrogen;

[0121] a linear or branched-C1-C6 alkylene-R8 group;

[0122] -C2-C6 alkenyl;

[0123] -X2-O-R7;

[0124]

[0125] -X2-NSO2-R7;

[0126] -C=C(R9)-Y1-O-R7;

[0127] a) C3-C6 cycloalkyl;

[0128] C3-C6 heterocycloalkyl optionally substituted by a hydroxy group;

[0129] C3-C6 cycloalkylene-Y2-R7;

[0130] C3-C6 heterocycloalkylene-Y2-R7 group, and

[0131] a heteroarylene-R7 group optionally substituted by a linear or branched C1-C6 alkyl group,

[0132] R7 represents a group selected from the group consisting of: a linear or branched C1-C6 alkyl group; (C3-C6) cycloalkylene-R8;

[0133]

[0134]

[0135] wherein Cy represents a C3-C8 cycloalkyl group,

[0136] R8 represents a group selected from the group consisting of: hydrogen; a straight or branched C1-C6 alkyl group, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R'c;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b 、-X'2-NR' a R' b 、-NR' c -X'2-N3 and:

[0137]

[0138] R9 represents a group selected from the group consisting of a linear or branched C1-C6 alkyl group, a trifluoromethyl group, a hydroxyl group, a halogen group, and a C1-C6 alkoxy group,

[0139] R 10 represents a radical selected from the group consisting of hydrogen, fluorine, chlorine, bromine, -CF3 and methyl,

[0140] R 11 represents a group selected from the group consisting of hydrogen, halogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8 and C3-C8 heterocycloalkylene-CH2-R8,

[0141] R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group,

[0142] R 14 and R 15 independently of one another represent hydrogen or a methyl group, or R 14 and R 15 Together with the carbon atoms that carry them, they form cyclohexyl groups,

[0143] R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0144] X1 represents a linear or branched C1-C4 alkylene group, which is optionally substituted by one or two groups selected from the group consisting of trifluoromethyl, hydroxy, halogen and C1-C6 alkoxy,

[0145] X2 represents a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from the group consisting of trifluoromethyl, hydroxy, halogen and C1-C6 alkoxy,

[0146] X'2 represents a linear or branched C1-C6 alkylene group,

[0147] R' a and R' brepresents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ;

[0148] C1-C6 alkylene-N + R' d R' e R' f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a hydroxyl group or a C1-C6 alkoxy group; and the group:

[0149]

[0150] or R' a and R' b Together with the nitrogen atoms that carry them, they form ring B3,

[0151] or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom that carries them,

[0152] R' c , R' d , R' e , R' f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0153] or R' d and R' e Together with the nitrogen atoms that carry them, they form ring B4,

[0154] or R' d , R' e and R' f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom that carries them,

[0155] Y1 represents a linear or branched C1-C4 alkylene group,

[0156] Y2 represents a bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-,

[0157] -CH2-SO2-, -C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-,

[0158] -NH-CO- or -NH-SO2-,

[0159] m=0, 1 or 2,

[0160] B1, B2, B3 and B4 independently represent C3-C8 heterocycloalkyl, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the group consisting of fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH2, oxo and piperidinyl,

[0161] wherein one of the R3, R8, and G groups (if present) is covalently attached to the linker, and wherein the valence of the atom is not exceeded by one or more substituents bonded thereto.

[0162] In some embodiments, for the ADC compound of Formula (I), D comprises a Bcl-xL inhibitor compound of Formula (I) or Formula (II) covalently attached to a linker L:

[0163]

[0164] or any of the foregoing enantiomers, diastereomers and / or addition salts thereof with a pharmaceutically acceptable acid or base (i.e., pharmaceutically acceptable salts), wherein:

[0165] R1 and R2 are independently selected from the group consisting of: hydrogen; linear or branched C1-C6 alkyl, which is optionally substituted by: hydroxyl or C1-C6 alkoxy groups; C3-C6 cycloalkyl; trifluoromethyl; linear or branched

[0166] C1-C6 alkylene-heterocycloalkyl, wherein the heterocycloalkyl group is optionally substituted with a straight-chain or branched C1-C6 alkyl group;

[0167] or R1 and R2 together with the carbon atom carrying them form a C3-C6 cycloalkylene group,

[0168] R3 represents a group selected from the following: hydrogen; C3-C6 cycloalkyl; linear or branched C1-C6 alkyl; -X1-NR a R b ;-X1-N + R a R bR c ;-X1-OR c ;-X1-COOR c ; -X1-PO(OH)2; -X1-SO2(OH); -X1-N3 and:

[0169]

[0170] R a and R b represents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR d R e ; C1-C6 alkylene-N + R d R e R f ; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a C1-C6 alkoxy group;

[0171] Group:

[0172]

[0173] or R a and R b Together with the nitrogen atom carrying them, they form ring B1;

[0174] or R a 、R b and R c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them,

[0175] R c 、R d 、R e 、R f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0176] or R d and R e Together with the nitrogen atoms that carry them, they form ring B2,

[0177] or R d 、R e and R f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them,

[0178] Het1 represents a group selected from the following:

[0179]

[0180] Het2 represents a group selected from the following:

[0181]

[0182] A1 is -NH-, -N(C1-C3 alkyl), O, S or Se,

[0183] A2 is N, CH or C(R5),

[0184] G is selected from the group consisting of:

[0185] -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、

[0186] -NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 、

[0187] C1-C6 alkyl, halogen, -NO2 and -CN optionally substituted with hydroxyl groups, wherein:

[0188] -R G1 and R G2 are independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms,

[0189] C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl;

[0190] -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; or

[0191] R G1 and R G2 Together with the atoms to which they are attached, they form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of:

[0192]

[0193] where R G4 is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl optionally substituted by 1 to 3 halogen atoms,

[0194] R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl group, a hydroxyl group or a methoxy group,

[0195] R5 represents a group selected from the following: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; C2-C6 alkenyl; C2-C6 alkynyl; halogen or -CN,

[0196] R6 represents a group selected from the following:

[0197] hydrogen;

[0198] -C2-C6 alkenyl;

[0199] -X2-O-R7;

[0200]

[0201] -X2-NSO2-R7;

[0202] -C=C(R9)-Y1-O-R7;

[0203] C3-C6 cycloalkyl;

[0204] C3-C6 heterocycloalkyl optionally substituted by a hydroxy group;

[0205] C3-C6 cycloalkylene-Y2-R7;

[0206] C3-C6 heterocycloalkylene-Y2-R7 group,

[0207] a heteroarylene-R7 group optionally substituted by a linear or branched C1-C6 alkyl group,

[0208] R7 represents a group selected from the following: linear or branched C1-C6 alkyl; (C3-C6) cycloalkylene-R8; or:

[0209]

[0210] wherein Cy represents a C3-C8 cycloalkyl group,

[0211] R8 represents a group selected from the following: hydrogen; linear or branched C1-C6 alkyl, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R' c ;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b 、-X'2-NR' a R' b、 -NR' c -X'2-N3 and:

[0212]

[0213] R9 represents a group selected from a linear or branched C1-C6 alkyl group, a trifluoromethyl group, a hydroxyl group, a halogen group, a C1-C6 alkoxy group,

[0214] R 10 represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl,

[0215] R 11 represents a group selected from hydrogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NRh R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8, C3-C8 heterocycloalkylene-CH2-R8,

[0216] R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group,

[0217] R 14 and R 15 independently of one another represent hydrogen or a methyl group, or R 14 and R 15 Together with the carbon atoms that carry them, they form cyclohexyl groups,

[0218] R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0219] X1 and X2 independently of one another represent a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy,

[0220] X'2 represents a linear or branched C1-C6 alkylene group,

[0221] R' a and R' b represents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ;

[0222] C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ; C1-C6 alkylene-N + R' d R' e R' f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted with a hydroxyl group or a C1-C6 alkoxy group;

[0223] Group:

[0224]

[0225] or R' a and R' b Together with the nitrogen atoms that carry them, they form ring B3,

[0226] or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them,

[0227] R' c , R' d , R' e , R' f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0228] or R' d and R' e Together with the nitrogen atoms that carry them, they form ring B4,

[0229] or R' d , R' e and R' f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them,

[0230] Y1 represents a linear or branched C1-C4 alkylene group,

[0231] Y2 represents bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-, -C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-,

[0232] m=0, 1 or 2,

[0233] p = 1, 2, 3 or 4,

[0234] B1, B2, B3 and B4 independently represent C3-C8 heterocycloalkyl, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH 2、 Oxo or piperidinyl,

[0235] wherein one of the R3 and R8 groups, if present, is covalently attached to the linker, and wherein the valence of the atom is not exceeded by one or more substituents bonded thereto; or

[0236]

[0237] or an enantiomer, diastereomer and / or a pharmaceutically acceptable acid or base addition salt thereof (i.e., a pharmaceutically acceptable salt), wherein:

[0238] n=0, 1 or 2,

[0239] ------ indicates a single bond or a double bond.

[0240] A4 and A5 independently represent a carbon or nitrogen atom,

[0241] Z1 represents a bond, -N(R)- or -O-, wherein R represents hydrogen or a linear or branched C1-C6 alkyl group,

[0242] R1 represents a group selected from the following: hydrogen; linear or branched C1-C6 alkyl, which is optionally substituted by: hydroxy or C1-C6 alkoxy; C3-C6 cycloalkyl; trifluoromethyl; linear or branched C1-C6 alkylene-heterocycloalkyl, wherein the heterocycloalkyl is optionally substituted by linear or branched C1-C6 alkyl;

[0243] R2 represents hydrogen or methyl;

[0244] R3 represents a group selected from the following: hydrogen; linear or branched C1-C4 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ;-X1-OR c ;-X1-COOR c ; -X1-PO(OH)2; -X1-SO2(OH); -X1-N3 and:

[0245]

[0246] R a and R b represents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O -; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR d R e ; C1-C6 alkylene-N + R d R e R f ; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a C1-C6 alkoxy group;

[0247] Group:

[0248]

[0249] or R a and R b Together with the nitrogen atom that carries them, they form ring B1;

[0250] or R a 、R b and R c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom that carries them,

[0251] R c 、R d 、R e 、R f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0252] or R d and R e Together with the nitrogen atoms that carry them, they form ring B2,

[0253] or R d 、R e and R f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom that carries them,

[0254] Het1 represents a group selected from the following:

[0255]

[0256] Het2 represents a group selected from the following:

[0257]

[0258] A1 is -NH-, -N(C1-C3 alkyl), O, S or Se,

[0259] A2 is N, CH or C(R5),

[0260] G is selected from the group consisting of:

[0261] -C(O)ORG3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、

[0262] -NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 、

[0263] C1-C6 alkyl, halogen, -NO2 and -CN optionally substituted with hydroxyl groups, wherein:

[0264] -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted with 1 to 3 halogen atoms, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl;

[0265] -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; or

[0266] R G1 and R G2 Together with the atoms to which they are attached, they form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of:

[0267]

[0268] where R G4 is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl optionally substituted by 1 to 3 halogen atoms,

[0269] R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl group, a hydroxyl group or a methoxy group,

[0270] R5 represents a group selected from the following: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; C2-C6 alkenyl; C2-C6 alkynyl; halogen or -CN,

[0271] R6 represents a group selected from the following:

[0272] hydrogen;

[0273] -C2-C6 alkenyl;

[0274] -X2-O-R7;

[0275]

[0276] -X2-NSO2-R7;

[0277] -C=C(R9)-Y1-O-R7;

[0278] C3-C6 cycloalkyl;

[0279] C3-C6 heterocycloalkyl optionally substituted by a hydroxy group;

[0280] C3-C6 cycloalkylene-Y2-R7;

[0281] C3-C6 heterocycloalkylene-Y2-R7 group,

[0282] a heteroarylene-R7 group optionally substituted by a linear or branched C1-C6 alkyl group,

[0283] R7 represents a group selected from the following: a linear or branched C1-C6 alkyl group;

[0284] (C3-C6)cycloalkylene-R8; or

[0285]

[0286]

[0287] wherein Cy represents a C3-C8 cycloalkyl group,

[0288] R8 represents a group selected from the following: hydrogen; linear or branched C1-C6 alkyl, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R'c;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b 、-X'2-NR' a R' b 、-NR' c -X'2-N3 and:

[0289]

[0290] R9 represents a group selected from a linear or branched C1-C6 alkyl group, a trifluoromethyl group, a hydroxyl group, a halogen group, a C1-C6 alkoxy group,

[0291] R 10 represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl,

[0292] R 11 represents a group selected from hydrogen, halogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and

[0293] -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8, C3-C8 heterocycloalkylene-CH2-R8,

[0294] R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group,

[0295] R 14 and R 15 independently represent hydrogen or methyl, or R14 and R 15 Together with the carbon atoms that carry them, they form cyclohexyl groups,

[0296] R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0297] X1 represents a straight or branched C1-C4 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy,

[0298] X2 represents a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy,

[0299] X'2 represents a linear or branched C1-C6 alkylene group,

[0300] R' a and R' b represents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ;

[0301] C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ;

[0302] C1-C6 alkylene-N + R' d R' e R' f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl,

[0303] wherein the phenyl group may be substituted by a hydroxyl group or a C1-C6 alkoxy group;

[0304] Group:

[0305]

[0306] or R' a and R' b Together with the nitrogen atoms that carry them, they form ring B3,

[0307] or R' a , R' b and R'c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them,

[0308] R' c , R' d , R' e , R' f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0309] or R' d and R' e Together with the nitrogen atoms that carry them, they form ring B4,

[0310] or R' d , R' e and R' f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them,

[0311] Y1 represents a linear or branched C1-C4 alkylene group,

[0312] Y2 represents bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-, -C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-,

[0313] m=0, 1 or 2,

[0314] p = 1, 2, 3 or 4,

[0315] B1, B2, B3 and B4 independently represent C3-C8 heterocycloalkyl, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH 2、 Oxo or piperidinyl,

[0316] wherein one of the R3 and R8 groups, if present, is covalently attached to the linker, and wherein the valency of the atom is not exceeded by one or more substituents bonded thereto.

[0317] In some embodiments, for Formula (I) or Formula (II), G is selected from the group consisting of:

[0318] -C(O)OR G3 、-C(O)NR G1 RG2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , halogen, -NO2 and -CN, among which:

[0319] -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted with 1 to 3 halogen atoms, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl;

[0320] -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; or

[0321] R G1 and R G2 Together with the atoms to which they are attached, they form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of:

[0322]

[0323] where R G4 is selected from the group consisting of a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group and a C3-C6 cycloalkyl group which are optionally substituted by 1 to 3 halogen atoms.

[0324] 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).

[0325] In some embodiments, the linker (L) comprises an attachment 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 specific embodiments, L comprises an attachment group; at least one bridging spacer group; and at least one cleavable group comprising a pyrophosphate group and / or a self-immolative group.

[0326] In some embodiments, the antibody-drug conjugate comprises a linker-drug (or "linker-load") moiety - (LD) having formula (A):

[0327]

[0328] where R 1 is an attachment group, L1 is a bridging spacer group, and E is a cleavable group.

[0329] In some embodiments, the cleavable group comprises a pyrophosphate group. In some embodiments, the cleavable group comprises:

[0330]

[0331] In some embodiments, the bridging spacer group comprises a polyoxyethylene (PEG) group. In some cases, the PEG group can be selected from PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, and PEG15. In some embodiments, the bridging spacer group can comprise: -CO-CH2-CH2-PEG12-. In other embodiments, the bridging spacer group comprises a butyryl, valeryl, hexanoyl, heptanoyl, or octanoyl group. In some embodiments, the bridging spacer group comprises a hexanoyl group.

[0332] In some embodiments, the attachment group is formed by 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 can have the following structure:

[0333]

[0334] An azido group may have the following structure: -N=N + =N - .

[0335] The cyclooctyne group can have the following structure:

[0336] and wherein -* is a bond to the antibody.

[0337] In some cases, the cyclooctyne group has the following structure: and wherein -* is a bond to the antibody.

[0338] In some embodiments, the attachment group has a , and wherein -* is a bond to the antibody.

[0339] In some embodiments, the antibody is linked to a linker (L) via an attachment group selected from:

[0340]

[0341] where -* is a bond to the antibody, and where is a bond to the bridging spacer group. As used herein, the term "joined" refers to covalently attached or covalently linked.

[0342] In some embodiments, the bridging spacer group is conjugated or covalently linked to the cleavable group.

[0343] In some embodiments, the bridging spacer group is -CO-CH2-CH2-PEG12-.

[0344] In some embodiments, the cleavable group is -pyrophosphate-CH2-CH2-NH2-.

[0345] In some embodiments, the cleavable group is conjugated or covalently linked to the Bcl-xL inhibitor (D).

[0346] In some embodiments, the linker comprises: an attachment group, at least one bridging spacer group, a peptide group, and at least one cleavable group.

[0347] In some embodiments, the antibody-drug conjugate comprises a linker-drug moiety, -(LD), having formula (B):

[0348]

[0349] where R 1 is an attachment 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 a Bcl-xL inhibitor. In some cases, m is 1 and the bridging spacer comprises:

[0350]

[0351] In some embodiments, at least one bridging 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 bridging spacer is selected from *-C(O)-CH2-CH2-PEG1-**, *-C(O)-CH2-PEG3-**, *-C(O)-CH2-CH2-PEG12**, *-NH-CH2-CH2-PEG1-**, polyhydroxyalkyl, *-C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)-**, *-C(O)-CH2-CH2-PEG12-NH-C(O)CH2-CH2-**, where ** represents the direct or indirect point of attachment of at least one bridging spacer to the attachment group, and * represents the direct or indirect point of attachment of at least one bridging spacer to the peptide group.

[0352] In some embodiments, L1 is selected from *-C(O)-CH2-CH2-PEG1-**, *-C(O)-CH2-PEG3-**, *-C(O)-CH2-CH2-PEG12**, *-NH-CH2-CH2-PEG1-**, and polyhydroxyalkyl, wherein ** represents L1 and R 1 , * represents the direct or indirect attachment point of L1 and Lp.

[0353] In some embodiments, m is 1 and L2 is -C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)-.

[0354] 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 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 comprise Val-Cit, Phe-Lys, Val-Ala, Val-Lys, Leu-Cit, Sulfo-Ala-Val-Cit, Sulfo-Ala-Val-Ala, Gly-Gly-Gly and / or Gly-Gly-Phe-Gly (SEQ ID NO: 68). In some embodiments, the peptide group (Lp) comprises In some embodiments, the peptide group (Lp) comprises the group:

[0355] In some cases, the peptide group comprises a group selected from:

[0356]

[0357] In some embodiments, the self-immolative group comprises p-aminobenzyl-carbamate, p-aminobenzyl-ammonium, p-amino-(sulfo)benzyl-ammonium, p-amino-(sulfo)benzyl-carbamate, p-amino-(alkoxy-PEG-alkyl)benzyl-carbamate, p-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-carbamate, or p-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-ammonium.

[0358] In some embodiments, m is 1 and the bridging spacer comprises

[0359] In some embodiments, the Linker-Drug Moiety-(LD) is formed from a compound selected from the group consisting of:

[0360]

[0361]

[0362]

[0363]

[0364]

[0365] In some embodiments, the antibody-drug conjugate comprises a linker-drug group, -(LD), comprising a formula selected from:

[0366]

[0367]

[0368]

[0369]

[0370]

[0371]

[0372] and wherein -* is a bond to the antibody.

[0373] In some embodiments, the antibody-drug conjugate comprises a linker drug group, -(LD), having formula (C):

[0374]

[0375] Where: R 1 is an attachment group, L1 is a bridging spacer; L p is a peptide group comprising 1 to 6 amino acids; D is a Bcl-xL inhibitor; G1-L2-A is a self-immolative spacer; L2 is a bond, methylene, neopentylene or C2-C3 alkenylene; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*,

[0376] Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and * of A represents the point of attachment to D; L3 is a spacer moiety; and R 2 It is the hydrophilic part.

[0377] In some embodiments, the antibody-drug conjugate comprises a linker drug group, -(LD), having formula (D):

[0378]

[0379] Where: R 1 is an attachment group; L1 is a bridging spacer; Lp is a peptide group comprising 1 to 6 amino acids; A is a bond, -OC(=O)-*,

[0380] -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or

[0381] -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, * of A represents the point of attachment to D; L3 is a spacer moiety; and R 2 It is the hydrophilic part.

[0382] In some embodiments, L1 comprises: or *-CH(OH)CH(OH)CH(OH)CH(OH)-**, wherein each n is an integer from 1 to 12, wherein * of L1 represents a direct or indirect point of attachment to Lp, and ** of L1 represents a point of attachment to R 1 direct or indirect attachment point.

[0383] In some embodiments, L1 is and n is an integer from 1 to 12, wherein * of L1 indicates a direct or indirect attachment point to Lp, and ** of L1 indicates a direct or indirect attachment point to R 1 direct or indirect attachment point.

[0384] In some embodiments, L1 is and n is 1, where * of L1 indicates the direct or indirect attachment point to Lp, and ** of L1 indicates the attachment point to R 1 direct or indirect attachment point.

[0385] In some embodiments, L1 is and n is 12, where * of L1 indicates the direct or indirect attachment point to Lp, and ** of L1 indicates the attachment point to R 1 direct or indirect attachment point.

[0386] In some embodiments, L1 is and n is an integer from 1 to 12, wherein * of L1 indicates a direct or indirect attachment point to Lp, and ** of L1 indicates a direct or indirect attachment point to R 1 direct or indirect attachment point.

[0387] In some embodiments, L1 comprises The * of L1 indicates the direct or indirect attachment point to Lp, and the ** of L1 indicates the direct or indirect attachment point to R 1 direct or indirect attachment point.

[0388] In some embodiments, L1 is a bridging spacer comprising:

[0389] *-C(=O)(CH2) m O(CH2) m -**;*-C(=O)((CH2) m O) t (CH2) n -**;*-C(=O)(CH2) m -**;

[0390] *-C(=O)NH((CH2) m O) t (CH2) n -**;

[0391] *-C(=O)O(CH2) m SSC(R 3 )2(CH2) m C(=O)NR 3 (CH2) m NR 3 C(=O)(CH2) m -**;

[0392] *-C(=O)O(CH2) m C(=O)NH(CH2) m -**;*-C(=O)(CH2) m NH(CH2) m -**;

[0393] *-C(=O)(CH2) m NH(CH2) n C(=O)-**;*-C(=O)(CH2) m X1(CH2) m -**;

[0394] *-C(=O)((CH2) m O) t (CH2) n X1(CH2)n -**;*-C(=O)(CH2) m NHC(=O)(CH2) n -**;

[0395] *-C(=O)((CH2) m O) t (CH2) n NHC(=O)(CH2) n -**;

[0396] *-C(=O)(CH2) m NHC(=O)(CH2) n X1(CH2) n -**;

[0397] *-C(=O)((CH2) m O) t (CH2) n NHC(=O)(CH2) n X1(CH2) n -**;

[0398] *-C(=O)((CH2) m O) t (CH2) n C(=O)NH(CH2) m -**;*-C(=O)(CH2) m C(R 3 )2-**or

[0399] *-C(=O)(CH2) m C(=O)NH(CH2) m -**, where * of L1 indicates the direct or indirect attachment point to Lp, and ** of L1 indicates the direct or indirect attachment point to R 1 Direct or indirect attachment point of

[0400] as well as

[0401] Each m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;

[0402] Each n is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; and

[0403] 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.

[0404] In some embodiments, R 2 The hydrophilic part includes polyethylene glycol, polyalkylene glycol, polyol, polysarcosine, sugar, oligosaccharide, polypeptide, 1 to 3 C2-C6 alkyl substituted with a group, or substituted with 1 to 2 groups independently selected from -OC(=O)NHS(O)2NHCH2CH2OCH3, -NHC(=O)C 1-4 In some embodiments, R2 is (where n is an integer between 1 and 6),

[0405] In some embodiments, the hydrophilic moiety comprises polyethylene glycol of the formula: Where R is H, -CH3CH2CH2NHC(=O)OR a 、-CH2CH2NHC(=O)R a or -CH2CH2C(=O)OR a , R' is OH, -OCH3, -CH2CH2NHC(=O)OR a 、-CH2CH2NHC(=O)R a or-OCH2CH2C(=O)OR a , and m and n are each an integer between 2 and 25 (e.g., between 3 and 25).

[0406] In some embodiments, the hydrophilic portion comprises

[0407] In some embodiments, the hydrophilic portion comprises polysarcosine, for example having the following moiety

[0408] wherein n is an integer between 3 and 25; and R is H, -CH3 or -CH2CH2C(=O)OH.

[0409] In some embodiments, L3 is a member having the structure The spacer portion,

[0410] in:

[0411] W is -CH2-, -CH2O-, -CH2N(R b )C(=O)O-、-NHC(=O)C(R b )2NHC(=O)O-、-NHC(=O)C(Rb )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-,

[0412] -S(O)2NH-, -NHS(O)2-, -C(=O)-, -C(=O)O-, or -NH-, wherein each R b independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl; and

[0413] X is a bond, triazolyl or -CH2-triazolyl-, wherein X and R 2 connect.

[0414] In some embodiments, L3 is a member having the structure The spacer portion,

[0415] in:

[0416] 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-,

[0417] -S(O)2NH-, -NHS(O)2-, -C(=O)-, -C(=O)O-, or -NH-, wherein each R b independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl; and

[0418] X is -CH2-triazolyl-C 1-4 Alkylene-OC(O)NHS(O)2NH-, -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 -, -CH2-triazolyl-C 1-4 Alkylene-OC(O)NHS(O)2NH-(CH2CH2O) n -or-C 4-6 Cycloalkylene-OC(O)NHS(O)2NH-(CH2CH2O) n -, wherein each n is independently 1, 2 or 3 and wherein X is attached to R 2 .

[0419] In some embodiments, the attachment group is formed by reacting a group comprising at least one reactive group. In certain instances, the attachment group is formed by reacting a first reactive group that is attached to a linker and a second reactive group that is attached to the antibody or is an amino acid residue of the antibody.

[0420] In some embodiments, at least one reactive group comprises:

[0421] thiols,

[0422] Maleimide,

[0423] Haloacetamide,

[0424] Azide,

[0425] Alkynes,

[0426] Cyclooctene,

[0427] Triarylphosphine,

[0428] Oxanorbornadiene,

[0429] Cyclooctyne,

[0430] Diaryltetrazine,

[0431] Monoaryl tetrazines,

[0432] Norbornene,

[0433] aldehyde,

[0434] Hydroxylamine,

[0435] Hydrazine,

[0436] NH2-NH-C(=O)-,

[0437] ketone,

[0438] Vinyl sulfone,

[0439] Aziridine,

[0440] amino acid residues,

[0441] -ONH2, -NH2,

[0442] -N3, -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, -C(O)NHNH2,

[0443]

[0444] in:

[0445] Each R 3 Independently selected from H and C1-C6 alkyl;

[0446] Each R 4 is 2-pyridyl or 4-pyridyl;

[0447] Each R 5 Independently selected from H, C1-C6 alkyl, F, Cl and -OH;

[0448] Each R 6 Independently selected from H, C1-C6 alkyl, F, Cl, -NH2, -OCH3, -OCH2CH3, -N(CH3)2, -CN, -NO2 and -OH;

[0449] Each R 7 Independently selected from H, C 1-6 Alkyl, fluorine, benzyloxy substituted by -C(=O)OH, benzyl substituted by -C(=O)OH, C 1-4 Alkoxy and C substituted by -C(=O)OH 1-4 alkyl.

[0450] In some embodiments, the first reactive group and the second reactive group include:

[0451] Thiol and maleimide,

[0452] Thiols and haloacetamides,

[0453] Thiol and vinyl sulfone,

[0454] Thiols and aziridines,

[0455] Azides and alkynes,

[0456] Azide and cyclooctyne,

[0457] Azide and cyclooctene,

[0458] Azide and triarylphosphine,

[0459] Azide and oxa-norbornadiene,

[0460] Diaryltetrazine and cyclooctene,

[0461] Monoaryl tetrazines and norbornene,

[0462] Aldehydes and hydroxylamines,

[0463] Aldehydes and hydrazines,

[0464] Aldehyde and NH2-NH-C(=O)-,

[0465] Ketones and hydroxylamines,

[0466] Ketones and hydrazines,

[0467] Ketone and NH2-NH-C(=O)-,

[0468] Hydroxylamine and

[0469] Amine and or

[0470] CoA or a CoA analog and a serine residue.

[0471] In some embodiments, the attachment group comprises a group selected from:

[0472]

[0473]

[0474]

[0475]

[0476] as well as

[0477] in:

[0478] R 32 It is H, C 1-4 Alkyl, phenyl, pyrimidine or pyridine;

[0479] R 35 It is H, C 1-6 Alkyl, phenyl or C substituted by 1 to 3 -OH groups 1-4 alkyl;

[0480] Each R 7 Independently selected from H, C 1-6 Alkyl, fluorine, benzyloxy substituted with –C(=O)OH, benzyl substituted with –C(=O)OH, C 1-4 Alkoxy and C substituted by –C(=O)OH 1-4 alkyl;

[0481] R 37 independently selected from H, phenyl and pyridine;

[0482] q is 0, 1, 2, or 3;

[0483] R 8 is H or methyl; and

[0484] R 9 It is H, -CH3 or phenyl.

[0485] 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 residue is selected from 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-Cit, Sulfo-Ala-Val-Ala, Gly-Gly-Gly and / or Gly-Gly-Phe-Gly (SEQ ID NO: 78).

[0486] In some embodiments, Lp is selected from:

[0487]

[0488] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0489] in:

[0490] R is H, -CH3 or -CH2CH2C(=O)OH;

[0491] A is a bond, -OC(=O)-*,

[0492] -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or

[0493] -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, with * in A representing the point of attachment to D; and

[0494] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0495]

[0496] wherein: -* is a bond to the antibody; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*; and R is -CH3 or -CH2CH2C(=O)OH.

[0497] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0498] in:

[0499] R is H, -CH3 or -CH2CH2C(=O)OH;

[0500] A is a bond, -OC(=O)-*,

[0501] -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or

[0502] -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, with * in A representing the point of attachment to D; and

[0503] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0504]

[0505] , wherein: -* is a bond to the antibody; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*; R is -CH3 or -CH2CH2C(=O)OH.

[0506] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula: in:

[0507] R is H, -CH3 or -CH2CH2C(=O)OH;

[0508] A is a bond, -OC(=O)-*,

[0509] -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or

[0510] -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, with * in A representing the point of attachment to D; and

[0511] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0512]

[0513] wherein: -* is a bond to the antibody; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*; and R is -CH3 or -CH2CH2C(=O)OH.

[0514] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0515] in:

[0516] Each R is independently selected from H, -CH3 and -CH2CH2C(=O)OH;

[0517] A is a bond, -OC(=O)-*, -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, with * in A representing the point of attachment to D; and

[0518] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0519]

[0520] wherein: -* is a bond to the antibody; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*; and R is -CH3 or -CH2CH2C(=O)OH.

[0521] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0522] in:

[0523] Each R is independently selected from H, -CH3 and -CH2CH2C(=O)OH;

[0524] A is a bond, -OC(=O)-*, -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, with * in A representing the point of attachment to D; and

[0525] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0526] wherein: -* is a bond to the antibody; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*; and R is -CH3 or -CH2CH2C(=O)OH.

[0527] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0528] in:

[0529] Xa is -CH2-, -OCH2-, -NHCH2-, or -NRCH2- and each R is independently H, -CH3, or -CH2CH2C(=O)OH;

[0530] A is a bond, -OC(=O)-*, -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, with * in A representing the point of attachment to D; and

[0531] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0532] wherein: -* is a bond to the antibody; and Xa, A, D, and R are as defined above. In some embodiments, Xa is -CH2- or -NHCH2-; A is a bond or -OC(=O)-*; and R is -CH3 or -CH2CH2C(=O)OH.

[0533] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0534] in:

[0535] R is H, -CH3 or -CH2CH2C(=O)OH;

[0536] A is a bond, -OC(=O)-*, -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, with * in A representing the point of attachment to D; and

[0537] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0538] wherein: -* is a bond to the antibody; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*; and R is -CH3 or -CH2CH2C(=O)OH.

[0539] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0540] in:

[0541] Xb is -CH2-, -OCH2-, -NHCH2-, or -NRCH2- and each R is independently H, -CH3, or -CH2CH2C(=O)OH;

[0542] A is a bond, -OC(=O)-*, -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, with * of A representing the point of attachment to D; and

[0543] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0544] wherein: -* is a bond to the antibody; and Xb, A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*; and R is -CH3 or -CH2CH2C(=O)OH.

[0545] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0546] in:

[0547] A is a bond, -OC(=O)-*, -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, with * of A representing the point of attachment to D; and

[0548] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0549] wherein: -* is a bond to the antibody; and A and are as defined above. In some embodiments, A is a bond or -OC(=O)-*.

[0550] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0551] in:

[0552] A is a bond, -OC(=O)-*, -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, with * in A representing the point of attachment to D; and

[0553] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0554] in:

[0555] -* is a bond to the antibody; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)-*.

[0556] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0557] in:

[0558] A is a bond, -OC(=O)-*, -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, with * in A representing the point of attachment to D; and

[0559] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0560] wherein: -* is a bond to the antibody; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)-*.

[0561] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0562] in:

[0563] A is a bond, -OC(=O)-*, -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, with * of A representing the point of attachment to D; and

[0564] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0565] wherein: -* is a bond to the antibody; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)-*.

[0566] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0567] in:

[0568] A is a bond, -OC(=O)-*, -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, with * of A representing the point of attachment to D; and

[0569] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0570] wherein: -* is a bond to the antibody; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)-*.

[0571] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0572] in:

[0573] A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(Ra )2C(R a )2N(CH3)C(=O)-*, where each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, with * in A representing the point of attachment to D; and

[0574] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0575] wherein: -* is a bond to the antibody; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)-*.

[0576] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0577] in:

[0578] A is a bond, -OC(=O)-*, -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, with * in A representing the point of attachment to D; and

[0579] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0580] wherein: -* is a bond to the antibody; and A and D are as defined above. In some embodiments, A is a bond or -OC(=O)-*.

[0581] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0582] in:

[0583] Each R is independently H, -CH3 or -CH2CH2C(=O)OH;

[0584] A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*、

[0585] Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and * of A represents the point of attachment to D; and

[0586] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0587] wherein: -* is a bond to the antibody; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*; and R is -CH3 or -CH2CH2C(=O)OH.

[0588] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0589] in:

[0590] Each R is independently H, -CH3 or -CH2CH2C(=O)OH;

[0591] A is a bond, -OC(=O)-*,

[0592] -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or

[0593] -OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*、

[0594] Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and * of A represents the point of attachment to D; and

[0595] D is a Bcl-xL inhibitor. In some embodiments, Linker-Drug Group-(LD) comprises the formula:

[0596] wherein: -* is a bond to the antibody; and A, D, and R are as defined above. In some embodiments, A is a bond or -OC(=O)-*;

[0597] R is -CH3 or -CH2CH2C(=O)OH.

[0598] In some embodiments, the Linker-Drug Group-(LD) comprises or is formed from a compound of the formula:

[0599] in:

[0600] A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*、

[0601] Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and * of A represents the point of attachment to D; and

[0602] D is a Bcl-xL inhibitor.

[0603] In some embodiments, A is a bond.

[0604] In some embodiments, A is -OC(=O)-*.

[0605] In some embodiments, R is -CH3.

[0606] In some embodiments, R is -CH2CH2COOH.

[0607] In some embodiments, the antibody-drug conjugate comprises a linker-drug group-(LD) formed from a compound selected from the group consisting of:

[0608]

[0609]

[0610]

[0611]

[0612]

[0613]

[0614]

[0615]

[0616]

[0617]

[0618]

[0619]

[0620]

[0621]

[0622]

[0623] In some embodiments, the antibody-drug conjugate comprises a linker-drug group, -(LD), comprising a formula selected from:

[0624]

[0625]

[0626]

[0627]

[0628]

[0629]

[0630]

[0631]

[0632]

[0633]

[0634]

[0635]

[0636]

[0637]

[0638] and wherein -* is a bond to the antibody.

[0639] In some embodiments, the Bcl-xL inhibitor (D) comprises a compound of Formula (I):

[0640]

[0641] or an enantiomer, diastereomer and / or pharmaceutically acceptable salt of any of the foregoing, wherein the variables are as described above for Formula (I). In some embodiments, R1 is a linear or branched C1-6 alkyl group and R2 is H.

[0642] In some embodiments, the Bcl-xL inhibitor (D) comprises a compound of formula (II):

[0643]

[0644] or any enantiomer, diastereomer, and / or pharmaceutically acceptable salt thereof, wherein the variables are as described above for formula (II). A1 and A5 both represent nitrogen atoms, R1 is a linear or branched C1-6 alkyl group; R2 is H; n is 1; and ------ represents a single bond.

[0645] In some embodiments, the Bcl-xL inhibitor (D) comprises a compound of Formula (IA) or (IIA):

[0646]

[0647] or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein:

[0648] Z1 represents a bond or -O-,

[0649] R3 represents a group selected from the following: hydrogen; C3-C6 cycloalkyl; linear or branched C1-C6 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ; and-X1-OR c ,

[0650] R a and R b represents, independently of one another, a group selected from the group consisting of: hydrogen; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl groups; and a C1-C6 alkylene group -SO2O - ,

[0651] R c represents hydrogen or a linear or branched C1-C6 alkyl group,

[0652] Het2 represents a group selected from the following:

[0653]

[0654] A1 is -NH-, -N(C1-C3 alkyl), O, S or Se,

[0655] A2 is N, CH or C(R5),

[0656] G is selected from the group consisting of:

[0657] -C(O)OH、-C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , C1-C6 alkyl optionally substituted by hydroxyl groups, halogen, -NO2 and -CN, wherein:

[0658] -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, and C1-C6 alkyl optionally substituted with 1 to 3 halogen atoms;

[0659] -R G3 is a C1-C6 alkyl group optionally substituted by 1 to 3 halogen atoms; or

[0660] RG1 and R G2 Together with the atoms to which they are attached, they form a C3-C8 heterocycloalkyl group;

[0661] R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl group, a hydroxyl group or a methoxy group,

[0662] R5 represents a group selected from the following: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; halogen or -CN,

[0663] R6 represents a group selected from the following:

[0664] -X2-O-R7; and

[0665] a heteroarylene-R7 group optionally substituted by a linear or branched C1-C6 alkyl group,

[0666] R7 represents a group selected from the following: a linear or branched C1-C6 alkyl group;

[0667] (C3-C6)cycloalkylene-R8; or

[0668]

[0669] wherein Cy represents a C3-C8 cycloalkyl group,

[0670] R8 represents a group selected from the following: hydrogen; linear or branched C1-C6 alkyl, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R' c ;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b ;-X'2-NR' a R' b :-NR' c -X'2-N3 and:

[0671]

[0672] R 10 represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl,

[0673] R 11 represents a group selected from hydrogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8, C3-C8 heterocycloalkylene-CH2-R8,

[0674] R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group,

[0675] R 14 and R 15 independently represent hydrogen or methyl, or R 14 and R 15 Together with the carbon atoms that carry them, they form cyclohexyl groups,

[0676] R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0677] X1 and X2 independently of one another represent a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy,

[0678] X'2 represents a linear or branched C1-C6 alkylene group,

[0679] R' a and R' b represents, independently of one another, a group selected from the group consisting of: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl group may be substituted by a linear or branched C1-C6 alkyl group; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ; C1-C6 alkylene-N + R' d R' e R' f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted with a hydroxyl group or a C1-C6 alkoxy group;

[0680] Group:

[0681]

[0682] or R' a and R' b Together with the nitrogen atoms that carry them, they form ring B3,

[0683] or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom that carries them,

[0684] R' c , R' d , R' e , R' f independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0685] or R' d and R' e Together with the nitrogen atoms that carry them, they form ring B4,

[0686] or R' d , R' e and R' f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom that carries them,

[0687] m=0, 1 or 2,

[0688] p = 1, 2, 3 or 4,

[0689] B3 and B4 independently represent a C3-C8 heterocycloalkyl group, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH 2、 Oxo or piperidinyl.

[0690] In some embodiments, for Formula (IA) or (IIA), G is selected from the group consisting of: -C(O)OH, -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NRG1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , halogen, -NO2 and -CN, among which:

[0691] -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, and C1-C6 alkyl optionally substituted with 1 to 3 halogen atoms;

[0692] -R G3 is a C1-C6 alkyl group optionally substituted by 1 to 3 halogen atoms; or

[0693] -R G1 and R G2 Together with the atoms to which they are attached, they form a C3-C8 heterocycloalkyl group.

[0694] In some embodiments, for Formula (I), (II), (IA), or (IIA), R7 represents a group selected from the group consisting of: linear or branched C1-C6 alkyl; (C3-C6)cycloalkylene-R8; or:

[0695]

[0696] wherein Cy represents a C3-C8 cycloalkyl group.

[0697] In some embodiments, for Formula (I), (II), (IA), or (IIA), R7 represents a group selected from:

[0698]

[0699] In some embodiments, the Bcl-xL inhibitor (D) comprises a compound of Formula (IB), (IC), (IIB), or (IIC):

[0700]

[0701] or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein:

[0702] For formula (IB) or (IC), R3 represents a group selected from the following: hydrogen; linear or branched C1-C6 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ; and-X1-OR c ;

[0703] For formula (IIB) or (IIC), Z1 represents a bond and R3 represents hydrogen; or Z1 represents -O- and R3 represents -X1-NR a R b ,

[0704] R a and R b represents, independently of one another, a group selected from the group consisting of: hydrogen; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl groups; and a C1-C6 alkylene group -SO2O - ,

[0705] R c represents hydrogen or a linear or branched C1-C6 alkyl group

[0706] R6 represents -X2-O-R7 or a heteroarylene-R7 group optionally substituted by a linear or branched C1-C6 alkyl group,

[0707] R7 represents a group selected from the following:

[0708]

[0709] R8 represents a group selected from the following: -NR' a R' b ;-O-X'2-NR' a R' b ; and -X'2-NR' a R' b ,

[0710] R 10 represents fluorine,

[0711] R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group,

[0712] R 14 and R 15 independently of one another represent hydrogen or a methyl group,

[0713] X1 and X2 independently of one another represent a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy,

[0714] X'2 represents a linear or branched C1-C6 alkylene group,

[0715] R' a and R' b independently of one another represent a group selected from the following: hydrogen; linear or branched C1-C6 alkyl optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-NR' d R' e ;

[0716] or R' a and R' b Together with the nitrogen atoms that carry them, they form ring B3,

[0717] R' d , R' e independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group,

[0718] B3 represents a C3-C8 heterocycloalkyl group, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spirocyclic system, (ii) contain one or two heteroatoms independently selected from oxygen and nitrogen in addition to nitrogen atoms, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl and oxo.

[0719] In some embodiments, R7 represents the following group:

[0720]

[0721] In some embodiments, R7 represents a group selected from:

[0722]

[0723] In some embodiments, for Formula (I), (IA), (IB), (IC), (II), (IIA), (IIB), or (IIC), R8 represents a group selected from:

[0724]

[0725] Wherein -* represents the bond to the linker.

[0726] In some embodiments, B3 represents a C3-C8 heterocycloalkyl group selected from pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, azepanyl, and 2,8-diazaspiro[4,5]decyl.

[0727] In some embodiments, D represents a Bcl-xL inhibitor attached to a linker L via a covalent bond, wherein the Bcl-xL inhibitor is selected from the compounds in Table A1:

[0728] Table A1

[0729]

[0730]

[0731]

[0732]

[0733]

[0734]

[0735]

[0736]

[0737]

[0738]

[0739]

[0740]

[0741] or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof.

[0742] In some embodiments, D comprises a formula selected from any one of the formulae in Table A2, or an enantiomer, diastereomer, and / or pharmaceutically acceptable salt of any of the foregoing.

[0743] Table A2

[0744]

[0745]

[0746]

[0747]

[0748]

[0749]

[0750]

[0751]

[0752]

[0753]

[0754]

[0755]

[0756] Wherein -* represents the bond to the linker.

[0757] In some embodiments, -(LD) is formed from a compound selected from Table B or an enantiomer, diastereomer, and / or pharmaceutically acceptable salt thereof. In some embodiments, the maleimide group in the compound of Table B Forms a covalent bond with an antibody or its antigen-binding fragment (Ab) to form a ADC compounds of formula (1) wherein * represents the point of attachment to Ab. For the compounds in Table A1, Table A2, Table B and Table 1, depending on their charge, these compounds may contain a pharmaceutically acceptable monovalent anionic counterion M1 - In some embodiments, the monovalent anionic counterion M1 - M1 can be selected from bromide, chloride, iodide, acetate, trifluoroacetate, benzoate, methanesulfonate, toluenesulfonate, trifluoromethanesulfonate, formate, etc. In some embodiments, the monovalent anionic counterion M1 - It is trifluoroacetate or formate.

[0758] Table B. Exemplary Linker Drug Groups

[0759]

[0760]

[0761]

[0762]

[0763]

[0764]

[0765]

[0766]

[0767]

[0768]

[0769]

[0770]

[0771]

[0772]

[0773]

[0774]

[0775]

[0776]

[0777]

[0778]

[0779]

[0780]

[0781]

[0782]

[0783]

[0784]

[0785]

[0786]

[0787]

[0788]

[0789]

[0790]

[0791]

[0792]

[0793]

[0794]

[0795]

[0796]

[0797]

[0798]

[0799]

[0800]

[0801]

[0802]

[0803]

[0804]

[0805]

[0806]

[0807]

[0808]

[0809]

[0810]

[0811]

[0812]

[0813]

[0814] In some embodiments, the antibody-drug conjugate has a formula according to any one of the structures shown in Table 1.

[0815] Table 1. Example ADC Structure

[0816]

[0817]

[0818] *Ab: Any antibody described herein (eg, EphA2-DANAPA antibody).

[0819] The ADC described above can also be expressed as follows:

[0820] Ab-(LD) p (1),

[0821] wherein Ab represents an anti-EphA2 antibody or antigenic fragment thereof covalently linked to the linker-load (L / P) described above; and 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 2. In some embodiments, p is 4. In some embodiments, p is determined by liquid chromatography-mass spectrometry (LC-MS).

[0822] 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 specific linker-drug disclosed herein, while the numbers "P#" and "C#" are used interchangeably to refer to a specific compound, unless otherwise indicated. 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, diastereomers, atropisomers, deuterated derivatives, and / or pharmaceutically acceptable salts of any of the foregoing.

[0823] In some embodiments, the present invention also provides a composition comprising multiple copies of an antibody-drug conjugate (e.g., any exemplary antibody-drug conjugate described herein). In some embodiments, the average p of the antibody-drug conjugate in the composition is about 2 to about 4.

[0824] In some embodiments, also provided herein are pharmaceutical compositions comprising an antibody-drug conjugate (e.g., any exemplary antibody-drug conjugate described herein) or a composition (e.g., any exemplary composition described herein), and a pharmaceutically acceptable carrier.

[0825] In some embodiments, the present invention further provides therapeutic uses of the ADC compounds and compositions, such as uses in the treatment of cancer. In some embodiments, the present disclosure provides methods for treating cancer (e.g., cancers expressing EphA2 antigens targeted by antibodies or antigen-binding fragments of ADCs). In some embodiments, the present disclosure provides methods for reducing or slowing the expansion of cancer cell populations in subjects. In some embodiments, the present disclosure provides methods for determining whether a subject suffering from or suspected of having cancer will respond to treatment with an ADC compound or composition disclosed herein.

[0826] An exemplary embodiment is a method of treating a subject having or suspected of having cancer, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein). In some embodiments, the cancer expresses the target antigen EphA2. 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, sarcoma, 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 (including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, gastric cancer, colon cancer, or head and neck cancer. In some embodiments, the cancer is lymphoma or gastric cancer. In some embodiments, the cancer is breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer, or colon cancer. In some embodiments, the cancer is breast cancer or non-small cell lung cancer.

[0827] Another exemplary embodiment is a method of reducing or inhibiting tumor growth in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein). In some embodiments, the tumor expresses the target antigen EphA2. 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, pancreatic cancer, gastric cancer, colon cancer, or spleen cancer. In some embodiments, the tumor is gastric cancer. In some embodiments, the tumor is breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer, or colon cancer. In some embodiments, the tumor is breast cancer or non-small cell lung cancer. In some embodiments, administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces or inhibits the growth of the tumor by at least about 10%, at least about 20%, at least about 30%, or 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%.

[0828] Another exemplary embodiment is a method of reducing or slowing the expansion of a cancer cell population in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein). In some embodiments, the cancer cell population expresses the target antigen EphA2.

[0829] In some embodiments, the cancer cell population is from a tumor or blood cancer. In some embodiments, the cancer cell population is from breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, 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 (including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, gastric cancer, colon cancer or head and neck cancer. In some embodiments, the cancer cell population is from a lymphoma or gastric cancer. In some embodiments, the cancer cell population is from breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer or colon cancer. In some embodiments, the cancer cell population is from breast cancer or non-small cell lung cancer. In some embodiments, administration of an 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 an 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%.

[0830] Another exemplary embodiment is an antibody-drug conjugate, composition or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition or pharmaceutical composition disclosed herein) for treating a subject having or suspected of having cancer. In some embodiments, the cancer expresses the target antigen EphA2. 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, sarcoma, 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 (including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, gastric cancer, colon cancer or head and neck cancer. In some embodiments, the cancer is lymphoma or gastric cancer. In some embodiments, the cancer cell population is from breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, stomach cancer, bladder cancer, or colon cancer. In some embodiments, the cancer cell population is from breast cancer or non-small cell lung cancer.

[0831] Another exemplary embodiment is the use of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein) in treating a subject suffering from or suspected of having cancer. In some embodiments, the cancer expresses the target antigen EphA2. 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, sarcoma, 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 (including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, gastric cancer, colon cancer, or head and neck cancer. In some embodiments, the cancer is lymphoma or gastric cancer. In some embodiments, the cancer is breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, stomach cancer, bladder cancer, or colon cancer. In some embodiments, the cancer is breast cancer or non-small cell lung cancer.

[0832] Another exemplary embodiment is the use of an antibody-drug conjugate, composition, or pharmaceutical composition (e.g., any exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein) in a method for preparing a medicament for treating a subject suffering from or suspected of having cancer. In some embodiments, the cancer expresses the target antigen EphA2. 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, sarcoma, 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 (including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, gastric cancer, colon cancer, or head and neck cancer. In some embodiments, the cancer is lymphoma or gastric cancer. In some embodiments, the cancer is breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer, or colon cancer. In some embodiments, the cancer is breast cancer or non-small cell lung cancer.

[0833] 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 exemplary antibody-drug conjugate, composition, or pharmaceutical composition disclosed herein) by providing a biological sample from the subject; contacting the sample with the 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 a target antigen. In some embodiments, the cancer expresses the target antigen EphA2. 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, sarcoma, 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 (including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, gastric cancer, colon cancer or head and neck cancer. In some embodiments, the cancer is lymphoma or gastric cancer. In some embodiments, the cancer is breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer or colon cancer. In some embodiments, the cancer is breast cancer or non-small cell lung cancer. In some embodiments, the sample is a tissue biopsy sample, a blood sample or a bone marrow sample.

[0834] Also disclosed are methods of producing the ADC compounds and compositions. An exemplary embodiment is a method of producing an antibody-drug conjugate by reacting an antibody or antigen-binding fragment with a cleavable linker that is conjugated or covalently attached to a Bcl-xL inhibitor under conditions that allow conjugation. BRIEF DESCRIPTION OF THE DRAWINGS

[0835] Figure 1 Shown are EBC-1 growth kinetics of EphA2-DANAPA-L11C-P25 ADC (30 mg / kg, SD, IV), 3207-DANAPA-L11C-P25 isotype control ADC (30 mg / kg, SD, IV), and EphA2-DANAPA CysMab control antibody (3.75 mg / kg, SD, IV) alone or in combination with paclitaxel (12.5 mg / kg, SD, IV).

[0836] Figure 2 Shown are the EBC-1 growth kinetics of different doses of EphA2-DANAPA-L11C-P25 ADC alone or in combination with paclitaxel.

[0837] Figure 3Shown are Panc03.27 growth kinetics for EphA2-DANAPA-L11C-P25 ADC, 3207-DANAPA-L11C-P25 isotype control ADC, and EphA2-DANAPA CysMab control antibody, alone or in combination with gemcitabine.

[0838] Figure 4 Shown are Panc03.27 growth kinetics for EphA2-DANAPA-L11C-P25 ADC, 3207-DANAPA-L11C-P25 isotype control ADC, and EphA2-DANAPA CysMab control antibody, alone or in combination with MAPK inhibitors LXH254 and CFF272.

[0839] Figure 5 The binding affinity of the EphA2 antibody to the HKB-11 cell line transduced to overexpress EphA2 is shown (round 1).

[0840] Figure 6 The binding affinity of the EphA2 antibody to the HKB-11 cell line transduced to overexpress EphA2 is shown (round 2).

[0841] Figure 7 The binding affinity of the EphA2 antibody to the HKB-11 cell line transduced to overexpress EphA2 is shown (round 3).

[0842] Figure 8 Figure 2 shows the binding kinetics of anti-EphA2 IgG antibody 1C1 and its light chain point mutant IgG to human EphA2 expressed on HKB11 cells.

[0843] Figure 9A and 9B Shown are the binding kinetics of the human, mouse, and cynomolgus monkey EphA2 extracellular domains to the anti-EphA2 IgG antibody 1C1 and its light chain point mutant IgG. DETAILED DESCRIPTION

[0844] The disclosed compositions and methods may be understood more readily by reference to the following detailed description taken in conjunction with the accompanying drawings, which form a part of this disclosure.

[0845] Herein, the description relates to compositions and methods of using the compositions. When the disclosure describes or claims a feature or embodiment associated with a composition, such feature or embodiment also applies to the method of using the composition. Similarly, when the disclosure describes or claims a feature or embodiment associated with a method of using the composition, such feature or embodiment also applies to the composition.

[0846] When expressing a range of values, it includes embodiments using any specific value within the range. In addition, references to values ​​specified in a range include each value within the range. All ranges are inclusive and combinable. When a value is expressed as an approximation by using the prefix "about," it will be understood that the specific value forms another embodiment. Unless the context clearly indicates otherwise, references to a specific numerical value include at least that specific value. Unless the specific context in which it is used provides otherwise, the use of "or" will mean "and / or." For all purposes, all references cited herein are incorporated by reference. When a reference conflicts with the specification, the specification will prevail.

[0847] Unless the context of the description indicates otherwise, e.g., when a structure or fragment of a structure is drawn without a symbol indicating a particular point of attachment, it can be used alone or attached to other components of the ADC, and it can do so in any orientation, e.g., an antibody attached to a chemical moiety, such as a linker-drug, at any suitable point of attachment. However, where indicated, the components of the ADC are attached in the orientation shown in a given formula. For example, if formula (1) is described as Ab-(LD) p , the group "-(LD)" is described as Then the detailed structure of formula (1) is It is not

[0848] It should be understood that certain features of the disclosed compositions and methods that are described herein in the context of separate embodiments for purposes of clarity may also be provided in combination in a single embodiment. Conversely, different features of the disclosed compositions and methods that are described in the context of a single embodiment for purposes of brevity may also be provided separately or in any subcombination.

[0849] As used throughout this application, antibody drug conjugates can be identified using a naming convention of the general format of "target antigen / antibody-linker-load". By way of example only, if an antibody drug conjugate is referred to as "target X-LO-P0", such conjugate will comprise an antibody that binds to target X, a linker designated as L0, and a load designated as P0. Alternatively, if an antibody drug conjugate is referred to as "anti-target X-LO-P0", such conjugate will comprise an antibody that binds to target X, a linker designated as L0, and a load designated as P0. In another alternative, if an antibody drug conjugate is referred to as "AbX-LO-P0", such conjugate will comprise an antibody designated as AbX, a linker designated as L0, and a load designated as P0. A control antibody drug conjugate comprising a nonspecific isotype control antibody may be referred to as "isotype control IgG1-LO-P0" or "IgG1-LO-P0".

[0850] 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 depicted by the formulas given herein except that one or more atoms are replaced by atoms 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, such as 3 H. 11 C, 13 C. 14 C. 15 N. 18 F and 36 Cl. Thus, it is to be understood that the present disclosure encompasses the incorporation of one or more of any of the foregoing isotopes (including, for example, radioisotopes (e.g. 3 H and 14 C)) or compounds wherein non-radioactive isotopes (such as 2 H and 13 C). Such isotope-labeled compounds can be used for metabolic studies (using 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 drug or substrate tissue distribution assays), or radiation therapy for patients. In particular, 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 skilled in the art, for example, by using an appropriate isotopically labeled reagent in place of the unlabeled previously used reagent.

[0851] definition

[0852] Throughout the specification and claims, various terms related to various aspects of this specification are used. Unless otherwise indicated, these terms should have their ordinary meanings in the art. Other specifically defined terms will be interpreted in a manner consistent with the definitions provided herein.

[0853] As used herein, unless the context clearly indicates otherwise, the singular forms "a, an," and "the" include plural forms. Unless otherwise indicated, the terms "comprising," "having," and "having a chemical formula" are to be interpreted as open terms (i.e., meaning "including but not limited to"). In addition, whenever "comprising" or another open term is used in an embodiment, it should be understood that the intermediate term "consisting essentially of" or the closed term "consisting of" may be used to more narrowly claim the same embodiment.

[0854] When used in the context of numerical values ​​and ranges, the term "about" or "approximately" refers to a value or range that is approximately or close to the value or range listed, so that the embodiment can be implemented as expected, as will be apparent to those skilled in the art from the teachings contained herein. In some embodiments, approximately refers to a numerical value ± 20%, 15%, 10%, 5%, 1%, 0.5% or 0.1%. In one embodiment, the term "about" refers to a value range that is 10% more or less than a specified value. In another embodiment, the term "about" refers to a value range that is 5% more or less than a specified value. In another embodiment, the term "about" refers to a value range that is 1% more or less than a specified value.

[0855] The terms "antibody-drug conjugate," "antibody conjugate," "conjugate," "immunoconjugate," and "ADC" are used interchangeably and refer to one or more therapeutic compounds (e.g., Bcl-xL inhibitors) linked to one or more antibodies or antigen-binding fragments. In some embodiments, an ADC is defined by the general formula: Ab-(LD) p (Formula 1), wherein Ab = antibody or antigen-binding fragment (e.g., anti-EphA2 antibody or antigen-binding fragment thereof), L = linker moiety, D = drug moiety (e.g., Bcl-xL inhibitor drug moiety), and p = number of drug moieties per antibody or antigen-binding fragment. In ADCs comprising a Bcl-xL inhibitor drug moiety, "p" refers to the number of Bcl-xL inhibitor compounds attached to the antibody or antigen-binding fragment.

[0856] The term "antibody" is used in the broadest sense to refer 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 foregoing, through at least one antigen recognition site within 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. A "complete" antibody is a glycoprotein that typically comprises 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 comprises 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 hypervariable regions, called complementarity determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged in the following order from amino terminus to carboxyl terminus: 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 region of the antibody can mediate the binding of the immunoglobulin 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 antibody can be a monoclonal antibody, a human antibody, a humanized antibody, a camelized antibody, or a chimeric antibody. 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 a complete antibody or an antigen-binding fragment thereof.

[0857] In some embodiments, antibodies or antibody fragments disclosed herein include modified or engineered amino acid residues, such as one or more cysteine ​​residues, as sites for conjugation to drug moieties (Junutula JR et al., Nat Biotechnol 2008, 26: 925-932). In one embodiment, the disclosure provides a modified antibody or antibody fragment comprising a substitution of one or more amino acids with cysteine ​​at a position described herein. The cysteine ​​substitution site is located in the constant region of the antibody or antibody fragment and is therefore suitable for use in a variety of antibodies or antibody fragments, and these sites are selected to provide stable and uniform conjugates. The modified antibody or fragment may have one, two or more cysteine ​​substitutions, and these substitutions may be used in combination with other modifications and conjugation methods as described herein. Methods for inserting cysteine ​​at specific positions of antibodies are known in the art, see, for example, 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 cysteine ​​at a position on its constant region selected from the group consisting of 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, and wherein these positions are numbered according to the EU system. In some embodiments, the modified antibodies or antibody fragments thereof comprise substitutions of one or more amino acids with cysteine ​​on their constant region selected from the following positions: 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, wherein positions are numbered according to the EU system, and wherein the light chain is a human kappa light chain. In certain embodiments, the modified antibodies or antibody fragments thereof comprise substitutions of two or more amino acids with cysteine ​​on their constant region, wherein the combination comprises substitutions 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, and wherein these 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, at position 152 of the antibody heavy chain, at position 360 of the antibody heavy chain, at position 107 of the antibody light chain, at position 165 of the antibody light chain, or at position 159 of the antibody light chain, and wherein the positions are numbered according to the EU system, and wherein the light chain is a kappa chain. In a specific embodiment, 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 at position 152 of the antibody heavy chain, wherein the positions are numbered according to the EU system. In a specific embodiment, the modified antibody or antibody fragment thereof comprises a substitution of one amino acid with cysteine ​​at position 360 of the antibody heavy chain, wherein the 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 cysteine ​​at position 107 of the antibody light chain, wherein the positions are numbered according to the EU system, and wherein the light chain is a kappa chain.

[0858] As used herein, the terms "antibody fragment" or "antigen binding fragment" or "functional antibody fragment" refer to at least a portion of an antibody that retains the ability to specifically interact with an antigen (e.g., EphA2) epitope (e.g., by binding, steric hindrance, stabilization / destabilization, spatial distribution). The antigen binding fragment may also retain the ability to be internalized into antigen-expressing cells. In some embodiments, the antigen binding fragment also retains immune effector activity. The terms antibody, antibody fragment, antigen binding fragment, etc. are intended to cover the use of binding domains from antibodies in the context of larger macromolecules, such as ADCs. It has been shown that fragments of full-length antibodies can perform the antigen binding function of full-length antibodies. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, scFv antibody fragments, disulfide-linked Fv (sdFv), Fd fragments consisting of VH and CH1 domains, linear antibodies, single domain antibodies such as sdAb (VL or VH), Camelidae VHH domains, multispecific antibodies formed from antibody fragments (e.g., a bivalent fragment comprising two Fab fragments linked by a disulfide bond 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 transplanted into scaffolds based on polypeptides such as type III fibronectin (Fn3) (see U.S. Patent No. 6,703,199, which describes fibronectin polypeptide miniantibodies). 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 chain and heavy chain variable regions are continuously connected, for example, via a synthetic linker, such as a short flexible polypeptide linker, and can be expressed as a single chain polypeptide, and wherein the scFv retains the specificity of the intact antibody from which it is derived. Unless otherwise specified, scFv can have VL and VH variable regions in any order (e.g., relative to the N-terminus and C-terminus of the polypeptide), and the scFv can comprise VL-linker-VH or can comprise VH-linker-VL. Antigen binding fragments are obtained using conventional techniques known to those skilled in the art, and the utility of the binding fragments (e.g., binding affinity, internalization) is screened in the same manner as for intact antibodies. For example, antigen binding fragments can be prepared by cleavage of intact proteins, for example, by protease or chemical cleavage.

[0859] As used herein, the term "complementarity determining region" or "CDR" refers to the amino acid sequence within the variable region of an antibody that confers antigen specificity and binding affinity. For example, typically, there are 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 of a number of well-known schemes, including those described in 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); or a combination thereof. 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, these CDRs correspond to amino acid residues defined as part of a Kabat CDR, and amino acid residues defined as part of a Chothia CDR. As used herein, CDRs defined according to the "Chothia" numbering scheme are sometimes also referred to as "hypervariable loops."

[0860] In some embodiments, under Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1) (e.g., one or more insertions after position 35), 50-65 (HCDR2), and 95-102 (HCDR3); the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1) (e.g., one or more insertions after position 27), 50-56 (LCDR2), and 89-97 (LCDR3). In some embodiments, under Chothia, the CDR amino acid residues in VH are numbered 26-32 (HCDR1) (e.g., one or more insertions after position 31), 52-56 (HCDR2), and 95-102 (HCDR3); the amino acid residues in VL are numbered 26-32 (LCDR1) (e.g., one or more insertions after position 30), 50-52 (LCDR2), and 91-96 (LCDR3). By combining the CDR definitions of both Kabat and Chothia, in some embodiments, the CDR comprises or consists of the following group: for example, amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in human VH or amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in human VL. In some embodiments, under IMGT, the CDR amino acid residues in VH are numbered as approximately 26-35 (CDR1), 51-57 (CDR2), and 93-102 (CDR3), and the CDR amino acid residues in VL are numbered as approximately 27-32 (CDR1), 50-52 (CDR2), and 89-97 (CDR3). In some embodiments, under IMGT, the program IMGT / DomainGap Align can be used to determine the CDR regions of an antibody.

[0861] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population, that is, the individual antibodies constituting the population are consistent except for possible naturally occurring mutations present in trace amounts. Monoclonal antibodies are highly specific, directed against a single antigenic epitope. In contrast, conventional (polyclonal) antibody preparations typically include multiple antibodies directed against (or specifically against) different epitopes. The modifier "monoclonal" represents the characteristics of an antibody obtained from a substantially homogeneous antibody population and should not be interpreted as requiring antibodies to be produced by any particular method. For example, the monoclonal antibodies to be used according to the present disclosure can be prepared by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). Monoclonal antibodies can also be isolated from phage antibody libraries using techniques such as those 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.

[0862] The monoclonal antibodies described herein can be non-human, human or humanized. The term specifically includes "chimeric" antibodies in which a portion of the heavy and / or light chain is identical or homologous to a corresponding sequence derived from an antibody of a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain or chains is identical or homologous to a corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they specifically bind to the target antigen and / or exhibit the desired biological activity.

[0863] 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. The term includes antibodies with variable regions, wherein the framework region and CDR region are derived from human sequences. In addition, if the antibody contains a constant region, the constant region is also derived from such human sequences, such as human germline sequences, or mutant forms of human germline sequences or antibodies containing a consensus framework sequence derived from human framework sequence analysis, for example, as described in Knappik et al., (2000), J Mol Biol; 296 (1): 57-86. The structure and position of immunoglobulin variable domains such as CDRs can be defined using a well-known numbering scheme, such as the Kabat numbering scheme, the Chothia numbering scheme, or a combination of Kabat and Chothia, and / or the ImMunoGenTics (IMGT) numbering scheme. The human antibodies of the present invention may include amino acid residues that are not encoded by human sequences (e.g., by random mutagenesis or site-specific mutagenesis in vitro, or by somatic mutation in vivo, or conservative substitution to introduce mutations to promote 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 another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0864] The term "recombinant human antibody" as used herein refers to a human antibody prepared, expressed, produced or isolated by recombinant means, such as an antibody isolated from an animal (e.g., a mouse) (the animal is transgenic or transchromosomal for human immunoglobulin genes) or a hybridoma prepared therefrom; an antibody isolated from a host cell transformed to express human antibodies (e.g., from a transfectoma); an antibody isolated from a recombinant combinatorial human antibody library; and an antibody prepared, expressed, produced or isolated by any other means (which involves splicing of all or part of human immunoglobulin genes, 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 can be subjected to in vitro mutagenesis (or, when a transgenic animal of a human Ig sequence is used, in vivo somatic mutagenesis is performed), and therefore, the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, although derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline library in vivo.

[0865] As used herein, the term "chimeric antibody" refers to an antibody in which the amino acid sequence of the immunoglobulin molecule is derived from two or more species. In some cases, the variable regions of the heavy and light chains correspond to those of an antibody derived from one species with the desired specificity, affinity, and activity, while the constant regions are homologous to antibodies derived from another species (e.g., humans) to minimize the immune response in the latter species.

[0866] As used herein, the term "humanized antibody" refers to an antibody form comprising sequences from non-human (e.g., mouse) antibodies and human antibodies. Such antibodies are chimeric antibodies of a type, which contain the minimum sequence derived from non-human immunoglobulin (NII). Generally, humanized antibodies will include substantially all of at least one and typically two variable domains, wherein all or substantially all of the hypervariable loops are corresponding to those of non-human immunoglobulin (NII), and all or substantially all of the framework (FR) regions are those of human immunoglobulin sequences. Humanized antibodies also optionally include at least a portion of an immunoglobulin constant region (Fc), typically at least a portion of a human immunoglobulin (HIG). Humanized antibodies can be further modified by replacing residues in the Fv framework region and / or in the non-human residues replaced to improve and optimize antibody specificity, affinity and / or activity.

[0867] As used herein, the term "Fc region" refers to a polypeptide comprising the CH3, CH2, and at least a portion of the hinge region of an antibody constant domain. Optionally, the Fc region may include a CH4 domain present in some antibody classes. The Fc region may 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 CH3 region of an antibody. In some embodiments, an antibody or antigen-binding fragment comprises the Fc region, CH1 region, and κ / λ region from an antibody constant domain. In some embodiments, an antibody or antigen-binding fragment comprises a constant region, such as a heavy chain constant region and / or a light chain constant region. In some embodiments, such a constant region is modified compared to a wild-type constant region. That is, a polypeptide may comprise alterations or modifications to one or more of the three heavy chain constant regions (CH1, CH2, or CH3) and / or the light chain constant region (CL). Exemplary modifications include additions, deletions, or substitutions of one or more amino acids in one or more domains. Such alterations may be included to optimize effector function, half-life, and the like.

[0868] As used herein with respect to an antibody or antigen-binding fragment, "internalization" refers to the ability of the antibody or antigen-binding fragment to pass through the lipid bilayer membrane of the cell into an internal compartment (i.e., "internalization") after binding to a cell, preferably into a degradation compartment in the cell. For example, an internalizing anti-EphA2 antibody is an antibody that can be taken into a cell after binding to EphA2 on the cell membrane. In some embodiments, the antibody or antigen-binding fragment used in the ADC disclosed herein targets a cell surface antigen (e.g., EphA2) and is an internalizing antibody or internalizing antigen-binding fragment (i.e., after antigen binding, the ADC transfers through the cell membrane). 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 into the cell by receptor-mediated endocytosis.

[0869] As used herein, "non-internalizing" with respect to an antibody or antigen-binding fragment refers to an antibody or antigen-binding fragment that remains on the cell surface after binding to a cell. In some embodiments, the antibody or antigen-binding fragment used in the ADC 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 and does not translocate across the cell membrane after antigen binding). In some embodiments, the non-internalizing antibody or antigen-binding fragment binds to a non-internalizing receptor or other cell surface antigen.

[0870] The terms "EPH receptor A2," "ephrin type A receptor 2," or "EphA2," as used herein, refer to any naturally occurring form of human EphA2. The term encompasses full-length human EphA2 (e.g., UniProt reference sequence: NP_004422.2; SEQ ID NO: 61), as well as any form of human EphA2 produced by cellular processing. The term also encompasses functional variants or fragments of human EphA2, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human EphA2 (i.e., variants and fragments are also encompassed unless the context indicates that the term is intended to refer only to the wild-type protein). EphA2 can be isolated from humans or produced recombinantly or synthetically.

[0871] As used herein, the term "anti-EphA2 antibody" or "antibody that binds to EphA2" refers to any form of an antibody or antigen-binding fragment thereof that binds (e.g., specifically binds) to EphA2. The term encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and biologically functional antigen-binding fragments, so long as they bind, e.g., specifically bind, to EphA2. WO2007 / 030642 provides exemplary EphA2 binding sequences, including exemplary anti-EphA2 antibody sequences, and is incorporated herein by reference. In some embodiments, the anti-EphA2 antibody used in the ADC disclosed herein is an internalizing antibody or internalizing antigen-binding fragment. 1C1 (WO2007 / 030642) is an exemplary anti-EphA2 antibody.

[0872] As used herein, the term "binding specificity" refers to the ability of an individual antibody or antigen-binding fragment to preferentially react with one antigenic determinant relative to different antigenic determinants. The degree of specificity indicates the degree to which an antibody or fragment preferentially binds to one antigenic determinant relative to different antigenic determinants. In addition, as used herein, the terms "specificity," "specific binding," and "specifically binds" refer to the binding reaction between an antibody or antigen-binding fragment (e.g., an anti-EphA2 antibody) and a target antigen (e.g., EphA2) from a heterogeneous population of proteins and other biological products. The binding specificity of an antibody can be tested by comparing binding to an appropriate antigen and binding to an unrelated antigen or antigen mixture under a given set of conditions. An antibody is considered specific if its binding affinity to the appropriate antigen is at least 2, 5, 7, 10, or more times greater than its affinity to an unrelated antigen or antigen mixture. A "specific antibody" or "target-specific antibody" is an antibody that binds only to the target antigen (e.g., EphA2) but does not bind (or exhibits minimal binding) to other antigens. In some embodiments, an antibody or antigen-binding fragment that specifically binds to a target antigen (e.g., EphA2) has a density of less than 1 x 10 -6 M, less than 1x10 -7 M, less than 1x10 -8 M, less than 1x10 -9 M, less than 1x10 -10 M, less than 1x10 -11 M, less than 1x10 -12 M, or less than 1x10 -13 M's K D In some embodiments, K D In some embodiments, K D Between 500 pM and 1 μM, 1 μM and 100 nM, or 100 mM and 10 nM.

[0873] As used herein, the term "affinity" refers to the strength of interaction between an antibody and an antigen at a single antigenic site. Without being bound by theory, within each antigen-binding site, the variable regions of the antibody "arms" interact with the antigen at multiple sites through weak non-covalent forces; the more interactions, the stronger the affinity, generally. The binding affinity of an antibody is the sum of the attractive and repulsive forces acting between the antigenic determinant and the antibody binding site.

[0874] The term "k on ” or “k a ” refers to the association rate constant for the association of an antibody with 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.

[0875] The term "k off ” or “k d ” refers to the dissociation rate constant for the dissociation of the antibody from the antibody / antigen complex. This rate can be determined using standard assays, such as surface plasmon resonance, biolayer interferometry, or ELISA assays.

[0876] The term "K D ” refers to the equilibrium dissociation constant of a specific antibody-antigen interaction. K D By k a / k d Calculation. The rate can be determined using standard assays, such as surface plasmon resonance, biolayer interferometry, or ELISA assays.

[0877] The term "epitope" refers to the part of an antigen that can be recognized and specifically bound by an antibody (or antigen binding fragment). Epitope determinants are typically composed of chemically active surface groups of molecules (e.g., amino acids or carbohydrates or sugar side chains) and may have specific three-dimensional structural characteristics as well as specific charge characteristics. When the antigen is a polypeptide, the epitope may be formed by continuous amino acids or non-continuous amino acids juxtaposed by the tertiary folding of the polypeptide. An epitope may be "linear" or "conformational." The difference between a conformational epitope and a linear epitope is that the binding to the former, rather than the latter, is lost in the presence of a denaturing solvent. The epitope to which an antibody (or antigen binding fragment) binds can be identified using any epitope mapping technique known in the art, including X-ray crystallography for epitope identification by direct observation of the antigen-antibody complex, and monitoring the binding of the antibody to the antigen fragment or mutant variant, or monitoring the solvent accessibility of different parts of the antibody and antigen. Exemplary strategies for mapping antibody epitopes include, but are not limited to, array-based oligopeptide 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).

[0878] Competitive binding and epitope binning can also be used to identify 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 the test antibody or binding protein reduces the binding of the reference antibody or binding protein to the target antigen, such as EphA2 (e.g., a binding protein comprising CDRs and / or variable domains identified in Tables CD) by at least about 50% (e.g., 50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.5% or more, or any percentage therebetween), and / or vice versa. In some embodiments, competitive binding may be due to shared or similar (e.g., partially overlapping) epitopes, or due to steric hindrance, where the antibody or binding protein binds at a nearby epitope (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 in a separate group of binding proteins that do not have overlapping or nearby epitopes.

[0879] As used herein, the terms "peptide," "polypeptide," and "protein" are used interchangeably to refer to polymers of amino acid residues. The term encompasses amino acid polymers comprising two or more amino acids joined to each other by peptide bonds, wherein one or more amino acid residues are artificial chemical mimetics of the corresponding naturally occurring amino acids, as well as applicable to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. The term includes, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, and the like. The term also includes natural peptides, recombinant peptides, synthetic peptides, or combinations thereof. Unless otherwise indicated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.

[0880] A "recombinant" protein refers to a protein (eg, an antibody) that is produced using recombinant techniques (eg, by expression of a recombinant nucleic acid).

[0881] An "isolated" protein is one that is free from at least some of the materials with which it is normally associated in its native 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 the living organism, is isolated. This definition includes production of antibodies in a variety of organisms and / or host cells known in the art.

[0882] As used herein, an "isolated antibody" is an antibody that has been identified and separated from one or more (e.g., a substantial portion) of the components (by weight) of its source environment, such as from components of a hybridoma cell culture or a different cell culture used for its production. In some embodiments, separation is performed to substantially remove components that might otherwise interfere with the suitability of the antibody for a desired application (e.g., for therapeutic use). Methods for preparing isolated antibodies are known in the art and include, but are not limited to, protein A chromatography, anion exchange chromatography, cation exchange chromatography, virus retention filtration, and ultrafiltration.

[0883] As used herein, the term "variant" refers to a nucleic acid sequence or amino acid sequence that is different from a reference nucleic acid sequence or amino acid sequence, but retains one or more biological properties of the reference sequence. The variant may contain one or more amino acid substitutions, deletions and / or insertions (or corresponding codon substitutions, deletions and / or insertions) relative to the reference sequence. The changes in the nucleic acid variant may not change the amino acid sequence of the 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 the amino acid sequence 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. The changes in the peptide variant sequence are generally limited or conservative, so the sequences of the unmodified peptide and the variant are very similar overall and are identical in many regions. In some embodiments, the peptide variant retains one or more functional properties of the unmodified peptide sequence. The variant and the unmodified peptide may differ in amino acid sequence due to one or more substitutions, additions, or deletions in any combination.

[0884] The variant of nucleic acid or peptide can be a naturally occurring variant or an unknown naturally occurring variant. Variants of nucleic acids and peptides can be prepared by mutagenesis techniques, by direct synthesis or by other techniques known in the art. Variants do not necessarily require physical manipulation of the reference sequence. As long as the sequence contains different nucleic acids or amino acids compared to the reference sequence, no matter how it is synthesized, it is considered to be a "variant". In some embodiments, the variant has a high sequence identity (i.e., 60% nucleic acid or amino acid sequence identity or higher) compared to the reference sequence. In some embodiments, peptide variants include polypeptides with amino acid substitutions, deletions and / or insertions, as 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 to the reference sequence or the corresponding fragment (e.g., functional fragment) of the reference sequence (e.g., variants that also retain one or more functions of the reference sequence). In some embodiments, nucleic acid variants encompass polynucleotides having amino acid substitutions, deletions and / or insertions, so long as the polynucleotides have 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 fragment (e.g., a functional fragment) of a reference sequence.

[0885] The term 'conservatively modified variant' applies to both amino acid and nucleic acid sequences. For nucleic acid sequences, conservatively modified variants refer to those nucleic acids that encode identical or substantially 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 each position where alanine is specified by a codon, the codon can be changed to any of the corresponding codons described without changing the encoded polypeptide. Such nucleic acid variations are "silent variations," which are one type of conservatively modified variation. Each nucleic acid sequence encoding a polypeptide herein also describes every possible silent variation of the nucleic acid. Those skilled in the art will recognize that every codon in a nucleic acid (except AUG, which is typically the only codon for methionine, and TGG, which is typically the only codon for tryptophan) can be modified to produce functionally identical molecules. Thus, every silent variation of a nucleic acid encoding a polypeptide is implicit in each described sequence. For polypeptide sequences, conservatively modified variants include a single substitution, deletion, or addition to the polypeptide sequence, whereby an amino acid is substituted with a chemically similar amino acid. Conservative substitutions providing functionally similar amino acids are well known in the art.

[0886] As used herein, the term "conservative sequence modification" refers to amino acid modifications that do not significantly affect or change, for example, the binding characteristics of 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 an antibody or antigen-binding fragment by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are substitutions in which an amino acid residue is replaced by an amino acid residue with a similar side chain. Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with the following side chains: 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), non-polar 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 within an antibody can be replaced with other amino acid residues from the same side chain family, and the altered antibodies can be tested using the functional assays described herein.

[0887] As used herein, the term "homologous" or "identity" refers to the subunit sequence identity between two polymeric molecules, such as between two nucleic acid molecules (such as two DNA molecules or two RNA molecules) or between two polypeptide molecules. When the subunit position in the two molecules is occupied by the same monomeric subunit; for example, if a position in each of the two DNA molecules is occupied by adenine, then they are homologous or identical at that position. The homology between two sequences is a direct function of the number of matching positions or homologous positions. For example, if half of the positions in the two sequences (e.g., five positions in a polymer that is ten subunits in length) are matched or homologous, then the two sequences are 50% homologous; if 90% of the positions (e.g., 9 out of 10) are matched or homologous, then the two sequences are 90% homologous.

[0888] By comparing two optimally aligned sequences over a comparison window, the percentage ratio of " sequence identity " is determined, wherein the fragment of the amino acid sequence in the comparison window can comprise an interpolation or deletion (for example, a gap or overhang) compared with a reference sequence (which does not comprise an interpolation or deletion) so that the two sequences are optimally aligned. This percentage ratio can be calculated by the following method: the number of positions at which identical amino acid residues occur in the two sequences is measured to produce a matching position number, the matching position number is divided by the total number of positions in the comparison window, and the result is multiplied by 100, thereby obtaining the sequence identity percentage ratio. The output is the identity percentage ratio of the subject sequence relative to the query sequence. The identity percentage ratio between the two sequences is a function of the number of identical positions that the two sequences have, and considering the number of gaps, the length of each gap, they need to be introduced to carry out the optimal comparison of the two sequences. In general, the amino acid identity or homology between the proteins disclosed herein, and variants thereof, including variants of the target antigen (e.g., EphA2) and variants of the antibody variable domain (including individual variant CDRs), is at least 80%, e.g., at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, nearly 100%, or 100% identical or homologous to the sequences described herein.

[0889] The comparison of sequences and the determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In some embodiments, the Needleman and Wunsch ((1970) J. Mol. Biol. 48: 444-53) algorithm, which has been incorporated into the GAP program in the GCG software package, is used to determine the percent identity between two amino acid sequences using a Blossum 62 matrix or a PAM250 matrix and 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 GAP program in the GCG software package is used to determine the percent identity between two nucleotide sequences using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. An exemplary set of parameters is the Blossum 62 scoring matrix with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5. The percent identity between two amino acid or nucleotide sequences can also be determined using the algorithm of Meyers and Miller ((1989) CABIOS 4:11-17), which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.

[0890] The term "pharmaceutical agent" is used herein to refer to a compound, a mixture of compounds, a biomacromolecule, an extract made from a biological material, or a combination of two or more thereof. The term "therapeutic agent" or "drug" refers to an agent that modulates a biological process and / or has biological activity. Bcl-xL inhibitors and ADCs containing them, as described herein, are exemplary therapeutic agents.

[0891] The term "chemotherapeutic agent" or "anticancer agent" is used herein to refer to all agents that are effective in treating cancer, regardless of their mechanism of action. Inhibition of metastasis or angiogenesis is generally a property of chemotherapeutic agents. Chemotherapeutic agents include antibodies, biomolecules, and small molecules, and encompass Bcl-xL inhibitors and ADCs containing them, as described herein. Chemotherapeutic agents can be cytotoxic agents or cytostatics. The term "cytostatic" 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 the expression activity and / or function of a cell.

[0892] As used herein, the term "extra large B-cell lymphoma" or "Bcl-xL" refers to any naturally occurring form of human Bcl-xL, an anti-apoptotic member of the Bcl-2 protein family. The term encompasses full-length human Bcl-xL (e.g., UniProt reference sequence: Q07817-1), as well as any form of human Bcl-xL that may be produced by cellular processing. The term also includes functional variants or fragments of human Bcl-xL, including but not limited to splice variants, allelic variants, and isoforms that retain one or more biological functions of human Bcl-xL (i.e., variants and fragments are also included unless the context indicates that the term is used to refer only to the wild-type protein). Bcl-xL can be isolated from humans or produced recombinantly or synthetically.

[0893] As used herein, the term "inhibit" means to reduce a biological activity or process by a measurable amount and may include but does not require complete prevention or inhibition. In some embodiments, "inhibit" means to reduce the expression and / or activity of Bcl-xL and / or one or more of its upstream regulators or downstream targets.

[0894] As used herein, the term "Bcl-xL inhibitor" refers to an agent that is capable of reducing the expression and / or activity of Bcl-xL and / or one or more of its upstream regulators or downstream targets. Exemplary Bcl-xL modulators, including exemplary Bcl-xL inhibitors, are described in WO2021 / 018858, WO2021 / 018857, WO2010 / 080503, WO2010 / 080478, WO2013 / 055897, WO2013 / 055895, WO2016 / 094509, WO2016 / 094517, WO2016 / 094505, Tao et al., ACS Medicinal Chemistry Letters (2014), 5(10), 1088-109, and Wang et al., ACS Medicinal Chemistry Letters (2020), 11(10), 1829-1836, each of which is incorporated herein by reference as exemplary Bcl-xL modulators, including exemplary Bcl-xL inhibitors, which can be included as drug moieties in the ADCs of the present disclosure.

[0895] As used herein, "Bcl-xL inhibitor drug moiety," "Bcl-xL inhibitor," and the like refer to a component of an ADC or composition that provides a Bcl-xL inhibitor compound or the structure of a compound modified for attachment to an ADC that retains substantially the same, similar, or enhanced biological function or activity compared to the original compound. In some embodiments, the Bcl-xL inhibitor drug moiety is component (D) in the ADC of formula (1).

[0896] The term "cancer" as used herein refers to the presence of cells with the typical characteristics of oncogenic cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and / or certain morphological characteristics. Typically, cancer cells can be in the form of a tumor or mass, but such cells can be present alone in a subject, or can circulate in the bloodstream as independent cells, such as leukemia or lymphoma cells. The term "cancer" includes all types of cancer and cancer metastasis, including hematological cancers, solid tumors, sarcomas, carcinomas and other solid and non-solid tumor cancers. Hematological cancers can include B cell malignancies, hematological cancers (leukemias), plasma cell cancers (myeloma, such as multiple myeloma) or lymph node cancers (lymphomas). Exemplary B cell malignancies include chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma and diffuse large B cell lymphoma. Leukemia can 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. The terms "acute lymphocytic leukemia" and "acute lymphocytic leukemia" can be used interchangeably to describe ALL. Lymphoma can include Hodgkin's lymphoma, non-Hodgkin's lymphoma, etc. Other blood cancers can include myelodysplastic syndrome (MDS). Solid tumors can include cancers, such as adenocarcinomas, such as breast cancer, pancreatic cancer, prostate cancer, colon cancer or colorectal cancer, lung cancer, gastric cancer, cervical cancer, endometrial cancer, ovarian cancer, bile duct cancer, glioma, melanoma, etc. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, 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 (including acute lymphocytic 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, prostate cancer, small cell lung cancer, spleen cancer or head and neck cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[0897] As used herein, the term "tumor" refers to any tissue mass resulting from excessive cell growth or proliferation, whether benign or malignant, including precancerous lesions. In some embodiments, the tumor is breast cancer, stomach 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 spleen cancer. In some embodiments, the tumor is stomach cancer.

[0898] The terms "tumor cell" and "cancer cell" are used interchangeably herein and refer to individual cells or populations of cells derived from a tumor or cancer, including non-tumorigenic cells and cancer stem cells. The terms "tumor cell" and "cancer cell" are modified by the term "non-tumorigenic" when referring only to those cells that lack the capacity to renew and differentiate, to distinguish these cells from cancer stem cells.

[0899] As used herein, the terms "target negative," "target antigen negative," or "antigen negative" refer to the absence of target antigen expression in 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 the target antigen can be described as target negative, while a cell or cell line that expresses the target antigen can be described as target positive.

[0900] 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.

[0901] As used herein, the term "a subject in need of treatment" refers to a subject who would benefit biologically, medically, or in quality of life from treatment (eg, treatment with any one or more of the exemplary ADC compounds described herein).

[0902] The term "treatment" as used herein refers to any improvement of any consequence of a disease, disorder or condition, such as prolonging survival, reducing morbidity and / or alleviating side effects, from the results of alternative treatment modalities. In some embodiments, treatment includes delaying or improving a disease, disorder or condition (i.e., slowing down or preventing or reducing the development of a disease or at least one clinical symptom thereof). In some embodiments, treatment includes delaying, alleviating or improving at least one physical parameter of a disease, disorder or condition, including those that the patient may not be able to discern. In some embodiments, treatment includes regulating the disease, disorder or condition physically (e.g., stabilization of discernible symptoms), physiologically (e.g., stabilization of physical parameters), or both. In some embodiments, treatment includes administering the ADC compound or composition to a subject (e.g., a patient) to obtain the therapeutic benefits listed herein. Treatment can be curing, healing, alleviating, delaying, preventing, alleviating, changing, remedying, improving, alleviating, improving or affecting a disease, disorder or condition (e.g., cancer), a symptom of a disease, disorder or condition (e.g., cancer), or susceptibility to 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.

[0903] As used herein, the term "prevention" or "prevention" of a disease, disorder, or condition refers to prophylactic treatment of the disease, disorder, or condition; or delaying the onset or progression of the disease, disorder, or condition.

[0904] As used herein, "pharmaceutical composition" refers to a formulation of a composition, such as an ADC compound or composition, and at least one other (and optionally more than one other) component suitable for administration to a subject, such as a pharmaceutically acceptable carrier, stabilizer, diluent, dispersant, suspending agent, thickener, and / or excipient. The pharmaceutical compositions provided herein are in a form that allows for administration and subsequent provision of the intended biological activity of the active ingredient and / or for achieving a therapeutic effect. The pharmaceutical compositions provided herein preferably do not contain additional components that are unacceptably toxic to the subject to which the formulation is to be administered.

[0905] As used herein, the terms "pharmaceutically acceptable carrier" and "physiologically acceptable carrier" are used interchangeably and refer to a carrier or diluent that does not cause significant irritation to the subject and does not eliminate the biological activity and properties of the administered ADC compound or composition and / or any other therapeutic agent in the composition. Pharmaceutically acceptable carriers can enhance or stabilize the composition or can be used to promote the preparation of the composition. Pharmaceutically acceptable carriers may 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 of ordinary skill in the art (see, e.g., Remington's Pharmaceutical Sciences, 18th edition, Mack Printing Company, 1990, pages 1289 to 1329). Unless any conventional carrier is incompatible with the active ingredient, its use in treatment or pharmaceutical compositions is considered. The carrier may be selected to minimize adverse side effects in the subject, and / or to minimize degradation of the active ingredient.An adjuvant may also be included in any of these formulations.

[0906] As used herein, the term "excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate the administration of an active ingredient. Formulations for parenteral administration may, for example, contain excipients such as sterile water or saline, polyalkylene glycols such as polyethylene glycol, vegetable oils, or hydrogenated naphthalene. Other exemplary excipients include, but are not limited to, calcium bicarbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, ethylene-vinyl acetate copolymer particles, and surfactants, including, for example, polysorbate 20.

[0907] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that does not abrogate the biological activity and properties of the compounds of the invention and does not cause significant irritation to a subject to which it is 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, and the like; 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, and the like; and (b) salts formed with elemental anions such as chloride, bromide, and iodide. 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 herein by reference.

[0908] In some embodiments, the antibody-drug conjugates (ADCs), linkers, payloads, and linker-payloads described herein may contain a monovalent anionic counterion M1, depending on their charge. - Any suitable anionic counterion may be used. In certain embodiments, the monovalent anionic counterion is a pharmaceutically acceptable monovalent anionic counterion. In certain embodiments, the monovalent anionic counterion M1 - M1 can be selected from bromide, chloride, iodide, acetate, trifluoroacetate, benzoate, methanesulfonate, toluenesulfonate, trifluoromethanesulfonate, formate, etc. In some embodiments, the monovalent anionic counterion M1 - It is trifluoroacetate or formate.

[0909] The term "therapeutically effective amount" or "therapeutically effective dose" as used herein refers to the amount of a compound as described herein (e.g., an ADC compound or composition as described herein) that is capable of achieving the desired therapeutic outcome (i.e., reduction or inhibition of improvement in enzyme or protein activity, improvement in symptoms, alleviation of symptoms or conditions, 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 or causes side effects, but only those side effects that are acceptable to the treating clinician in view of the patient's condition. In some embodiments, a therapeutically effective amount is effective for detectably killing, reducing and / or inhibiting the growth or spread of cancer cells, the size or number of tumors, and / or other measurements of the level, stage, progression and / or severity of cancer. The term also applies to doses that induce a specific response in target cells, such as a reduction, slowing or inhibition of cell growth. A therapeutically effective amount can be determined by first administering a low dose and then incrementally increasing the dose until the desired effect is achieved. The therapeutically effective amount may also vary depending on the intended application (in vitro or in vivo) or the subject and disease condition being treated, such as the subject's weight and age, the severity of the disease condition, the mode of administration, etc., which can be readily determined by one of ordinary skill in the art. The specific amount may vary depending on, for example, the specific pharmaceutical composition, the subject's age and existing health condition or risk of health condition, the dosing regimen to be followed, the severity of the disease, whether to administer in combination with other drugs, the timing of administration, the tissue to which it is administered, and the physical delivery system to 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 down or stop) tumor metastasis, inhibit (e.g., slow down or stop) tumor growth, and / or alleviate one or more symptoms.

[0910] The term "prophylactically effective amount" or "prophylactically effective dose" as used herein refers to an amount of a compound disclosed herein (e.g., an ADC compound or composition described herein) effective to achieve the desired prophylactic effect at the necessary dosage and time period. Typically, a prophylactic dose will be less than a therapeutically effective amount because the prophylactic dose is used in subjects before or at an early stage of disease. In some embodiments, a prophylactically effective amount can prevent the onset of disease symptoms, including symptoms associated with cancer.

[0911] 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 comprising a Bcl-xL inhibitor drug moiety, "p" refers to the number of Bcl-xL inhibitor compounds attached to the antibody or antigen-binding fragment. For example, if two Bcl-xL inhibitor compounds are attached to the antibody or antigen-binding fragment, then 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."

[0912] Antibody-drug conjugates

[0913] Antibody-drug conjugate (ADC) compounds disclosed herein include those with anticancer activity. Specifically, ADC compounds include antibodies or antigen-binding fragments conjugated (i.e., covalently attached via a linker) to a drug moiety (e.g., a Bcl-xL inhibitor), wherein 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 Bcl-xL and / or one or more upstream regulators or downstream targets thereof. Without being bound by theory, in some embodiments, by targeting Bcl-xL expression and / or activity, the ADCs disclosed herein can provide effective anticancer agents. Moreover, without being bound by theory, by conjugating the drug moiety to an antibody that binds to an antigen associated with expression in tumor cells or cancer, the ADC can provide improved activity, better cytotoxic specificity, and / or reduced off-target killing, as compared to the drug moiety when administered alone.

[0914] Thus, in some embodiments, the components of the ADC are selected to (i) retain one or more therapeutic properties exhibited by the antibody and drug moiety alone, (ii) maintain the specific binding properties of the antibody or antigen-binding fragment; (iii) optimize drug loading and drug-antibody ratio; (iv) allow for delivery (e.g., intracellular delivery) of the drug moiety via stable attachment to the antibody or antigen-binding fragment; (v) maintain the stability of the ADC as a complete conjugate until transport or delivery to the target site; (vi) minimize aggregation of the ADC before or after administration; (vii) allow for the therapeutic effect (e.g., cytotoxic effect) of the drug moiety following cleavage or other release mechanisms in the cellular environment; (viii) exhibit in vivo anti-cancer therapeutic efficacy similar to or superior to that of the isolated antibody and drug moiety; (ix) minimize off-target killing caused by the drug moiety; and / or (x) exhibit desired pharmacokinetic and pharmacodynamic properties, formulation, and toxicological / immunological characteristics. Each of these properties can provide improved ADCs for therapeutic use (Ab et al. (2015) Mol Cancer Ther. 14: 1605-13).

[0915] The ADC compounds disclosed herein can selectively deliver effective doses of cytotoxic agents or cytostatic agents 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 cell. In some embodiments, the disclosed ADCs are particularly effective in 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 on cancer cells that do not express the target antigen.

[0916] In certain aspects, provided herein are ADC compounds comprising an anti-EphA2 antibody or antigen-binding fragment thereof (Ab), a Bcl-xL inhibitor drug moiety (D), and a linker moiety (L) that covalently attaches the Ab to D. In some embodiments, provided herein are ADC compounds comprising an antibody or antigen-binding fragment thereof (Ab) that targets cancer cells, a Bcl-xL inhibitor drug moiety (D), and a linker moiety (L) that covalently attaches the Ab to D. In some embodiments, the antibody or antigen-binding fragment is capable of binding to a tumor-associated antigen (e.g., EphA2), for example, with high specificity and high affinity. In some embodiments, the antibody or antigen-binding fragment is internalized into the target cell upon binding, for example, into a degradation compartment within the cell. In some embodiments, the ADC is internalized upon binding to the target cell, undergoes degradation, and releases the Bcl-xL inhibitor drug moiety to kill the cancer cell. The Bcl-xL 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.

[0917] An exemplary ADC has the formula (1):

[0918] Ab-(L-D) p (1)

[0919] where Ab = an anti-EphA2 antibody or antigen-binding fragment, L = a linker moiety, D = a Bcl-xL inhibitor drug moiety, and p = the number of Bcl-xL inhibitor drug moieties / antibody or antigen-binding fragment.

[0920] A. Antibody

[0921] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment (Ab) of formula (1) specifically binds to a target antigen on a cell. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment (Ab) of formula (1) specifically binds to a target antigen on a cancer cell. In some embodiments, the cell or the cancer cell expresses EphA2. In some embodiments, the target antigen EphA2 has the following amino acid sequence:

[0922] <NCBI Reference Sequence: NP_004422.2>

[0923]

[0924] The anti-EphA2 antibody or antigen-binding fragment can bind to the target antigen with a dissociation constant (K D ) of ≤1 mM, ≤100 nM, or ≤10 nM or any amount in between, as measured by, for example, analysis. In some embodiments, K D is from 1 pM to 500 pM. In some embodiments, K D is between 500 pM and 1 μM, 1 μM and 100 nM, or 100 nM and 10 nM.

[0925] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment is a tetrameric anti-EphA2 antibody (also referred to as an immunoglobulin or a full-length or intact antibody) that includes two heavy chains and two light chains. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment is an antigen-binding fragment of an immunoglobulin against EphA2. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment is an antigen-binding fragment of an immunoglobulin against EphA2 that retains the ability to bind to the target cancer antigen and / or provides at least one function of an immunoglobulin.

[0926] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment is an internalizing anti-EphA2 antibody or internalizing anti-EphA2 antigen-binding fragment thereof. In some embodiments, the internalizing anti-EphA2 antibody or internalizing anti-EphA2 antigen-binding fragment thereof binds to a target cancer antigen expressed on the surface of a cell and enters the cell after binding. In some embodiments, after the ADC enters and is present in a cell expressing the target cancer antigen (i.e., after the ADC is internalized), the Bcl-xL inhibitor drug portion of the ADC is released from the anti-EphA2 antibody or antigen-binding fragment of the ADC, for example, by cleavage, by degradation of the antibody or antigen-binding fragment, or by any other suitable release mechanism. In some embodiments, the cancer expresses EphA2.

[0927] In some embodiments, the anti-EphA2 antibody comprises a mutation that mediates reduced or no antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC). In some embodiments, these mutations are referred to as Fc silence, Fc silence, or Fc silent mutations. In some embodiments, amino acid residues L234 and L235 of the IgG1 constant region are substituted with A234 and A235 (also referred to as "LALA"). In some embodiments, amino acid residue N297 of the IgG1 constant region is substituted with A297 (also referred to as "N297A"). In some embodiments, amino acid residues D265 and P329 of the IgG1 constant region are substituted with A265 and A329 (also referred to as "DAPA"). Other antibody Fc silent mutations can also be used. In some embodiments, Fc silent mutations, such as D265A, N297A, and P329A (also referred to as "DANAPA"), are used in combination.

[0928] The amino acid sequences of exemplary anti-EphA2 antibodies of the disclosure are listed in Tables C and D.

[0929] As described herein, if anti-EphA2 antibodies are modified, they are further designated by the modification. For example, if selected amino acids in anti-EphA2 antibodies have been changed to cysteine ​​(e.g., E152C, S375C according to EU numbering of the antibody heavy chain to facilitate conjugation with a linker-drug moiety), they are designated as "CysMab"; or, if anti-EphA2 antibodies have been modified with Fc silent mutations D265A, N297A, and P329A in the IgG1 constant region according to EU numbering, "DANAPA" is added to the antibody name. If anti-EphA2 antibodies are used in antibody-drug conjugates, they are named using the following format: Antibody Name-Linker-Patent.

[0930] Table C. Amino acid sequences of mAb CDRs and variable regions

[0931]

[0932]

[0933]

[0934]

[0935]

[0936]

[0937]

[0938]

[0939]

[0940]

[0941] Table D. Amino acid and nucleic acid sequences of full-length mAb IgG chains

[0942]

[0943]

[0944]

[0945]

[0946]

[0947]

[0948]

[0949]

[0950]

[0951]

[0952]

[0953] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment of the ADC disclosed herein may comprise any of the heavy and light chain variable domain sets listed in the above table or a set of six CDRs from any of the heavy and light chain variable domain sets listed in the above table. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment of the ADC disclosed herein may comprise a conservatively modified amino acid sequence and / or an amino acid sequence homologous to the sequences listed in the above table, as long as the ADC retains the ability to bind to its target cancer antigen (e.g., K D Less than 1x10 -8 M) and retain one or more functional properties of the ADCs disclosed herein (e.g., the ability to internalize, bind to an antigen target, such as an antigen expressed on a tumor or other cancer cell, etc.).

[0954] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment of the ADC disclosed herein further comprises human heavy and light chain constant domains or fragments thereof. For example, the anti-EphA2 antibody or antigen-binding fragment of the ADC may comprise a human IgG heavy chain constant domain (e.g., IgG1) and a human kappa or lambda light chain constant domain. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment of the ADC comprises a human immunoglobulin G subtype 1 (IgG1) heavy chain constant domain and a human Ig kappa light chain constant domain.

[0955] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:2, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:3, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:12, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:14, SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:32, SEQ ID NO:35, or SEQ ID NO:70.

[0956] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:2, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:3, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:12, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:14.

[0957] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:2, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:3, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:12, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:26.

[0958] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:2, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:3, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:12, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:29.

[0959] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:2, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:3, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:12, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:32.

[0960] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:2, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:3, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:12, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:35.

[0961] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:2, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:3, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:12, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:70.

[0962] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:17, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:33, or SEQ ID NO:36.

[0963] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:17.

[0964] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:24.

[0965] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:27.

[0966] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:30.

[0967] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:33.

[0968] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:5, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:36.

[0969] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:8, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:18, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:17, SEQ ID NO:24, SEQ ID NO:27, SEQ ID NO:30, SEQ ID NO:33, or SEQ ID NO:36.

[0970] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:8, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:18, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:17.

[0971] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:8, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:18, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:24.

[0972] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:8, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:18, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:27.

[0973] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:8, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:18, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:30.

[0974] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:8, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:18, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:33.

[0975] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO:7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO:8, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO:9; a light chain CDR1 (LCDR1) consisting of SEQ ID NO:18, a light chain CDR2 (LCDR2) consisting of SEQ ID NO:13, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO:36.

[0976] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 10, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 17, SEQ ID NO: 24, SEQ ID NO: 27, SEQ ID NO: 30, SEQ ID NO: 33, or SEQ ID NO: 36.

[0977] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 10, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 17.

[0978] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 10, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 24.

[0979] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 10, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 27.

[0980] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 10, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 30.

[0981] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 10, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 33.

[0982] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises the following three heavy chain CDRs and three light chain CDRs: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 10, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 6, and a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 4; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 15, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 16, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 36.

[0983] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 11. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 11, or a sequence that is at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 11.

[0984] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 19. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 19, or a sequence that is at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 19.

[0985] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 20. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 20, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 20.

[0986] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 21. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 21, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 21.

[0987] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 22. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 22, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 22.

[0988] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 23. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 23, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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.

[0989] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 25. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 25, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 25.

[0990] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 28. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 28, or a sequence that is at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 28.

[0991] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 31. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 31, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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.

[0992] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 34. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 34, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 34.

[0993] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 71. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 71, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 71.

[0994] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 72, or a sequence having at least 95% identity to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 72.

[0995] In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 73. In some embodiments, the anti-EphA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region amino acid sequence of SEQ ID NO: 1 and a light chain variable region amino acid sequence of SEQ ID NO: 73, or a sequence that is at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 1 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: 73.

[0996] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO:37, or a sequence at least 95% identical to SEQ ID NO:37, and a light chain amino acid sequence of SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77, or a sequence at least 95% identical to SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO:37 and a light chain amino acid sequence of SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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:37 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:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77.

[0997] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 41, or a sequence at least 95% identical to SEQ ID NO: 41. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 41, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37 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: 41.

[0998] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 43, or a sequence at least 95% identical to SEQ ID NO: 43. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 43, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37 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: 43.

[0999] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 45, or a sequence at least 95% identical to SEQ ID NO: 45. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 45, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 45.

[1000] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 47, or a sequence at least 95% identical to SEQ ID NO: 47. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 47, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 47.

[1001] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 49, or a sequence at least 95% identical to SEQ ID NO: 49. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 49, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 49.

[1002] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 51, or a sequence at least 95% identical to SEQ ID NO: 51. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 51, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 51.

[1003] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 53, or a sequence at least 95% identical to SEQ ID NO: 53. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 53, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 53.

[1004] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 55, or a sequence at least 95% identical to SEQ ID NO: 55. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 55, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 55.

[1005] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 57, or a sequence at least 95% identical to SEQ ID NO: 57. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 57, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 57.

[1006] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 59, or a sequence at least 95% identical to SEQ ID NO: 59. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 59, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 59.

[1007] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 75, or a sequence at least 95% identical to SEQ ID NO: 75. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 75, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 75.

[1008] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 76, or a sequence at least 95% identical to SEQ ID NO: 76. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 76, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 76.

[1009] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, or a sequence at least 95% identical to SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 77, or a sequence at least 95% identical to SEQ ID NO: 77. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 37, and a light chain amino acid sequence of SEQ ID NO: 77, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 37, 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: 77.

[1010] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO:39, or a sequence at least 95% identical to SEQ ID NO:39, and a light chain amino acid sequence of SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77, or a sequence at least 95% identical to SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO:39 and a light chain amino acid sequence of SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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:39 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:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77.

[1011] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 39, or a sequence at least 95% identical to SEQ ID NO: 39, and a light chain amino acid sequence of SEQ ID NO: 41, or a sequence at least 95% identical to SEQ ID NO: 41. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 39, and a light chain amino acid sequence of SEQ ID NO: 41, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 39, 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: 41.

[1012] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO:74, or a sequence at least 95% identical to SEQ ID NO:74, and a light chain amino acid sequence of SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77, or a sequence at least 95% identical to SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO:74 and a light chain amino acid sequence of SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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:74 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:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:76, or SEQ ID NO:77.

[1013] In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 74, or a sequence at least 95% identical to SEQ ID NO: 74, and a light chain amino acid sequence of SEQ ID NO: 41, or a sequence at least 95% identical to SEQ ID NO: 41. In some embodiments, the anti-EphA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 74, and a light chain amino acid sequence of SEQ ID NO: 41, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-EphA2 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: 74, 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: 41.

[1014] Residues in two or more polypeptides are said to "correspond" if the residues occupy similar positions in the polypeptide structure. Similar positions in two or more polypeptides can be determined by aligning the polypeptide sequences based on amino acid sequence or structural similarity. Those skilled in the art will appreciate that it may be necessary to introduce gaps in either sequence to produce a satisfactory alignment.

[1015] In some embodiments, amino acid substitution is of a single residue. Insertion is generally about 1 to about 20 amino acid residues, but as long as biological function (for example, in combination with the target antigen) is retained, sizable insertion can be tolerated. Disappearance is generally within the scope of about 1 to about 20 amino acid residues, although in some cases deletion may be much larger. Replacement, disappearance, insertion or its any combination can be used to obtain final derivative or variant. Usually these changes are carried out on a minority of amino acids, thereby with the change of molecule, particularly the immunogenicity and specificity of antigen-binding proteins are minimized. However, in some cases, larger changes can be tolerated. Conservative substitution can be carried out according to the chart shown in Table 2.

[1016] Table 2

[1017]

[1018]

[1019] In some embodiments where variant antibody sequences are used in ADCs, the variants generally exhibit the same qualitative biological activity and will elicit the same immune response, although variants can also be selected to modify the characteristics of the antigen binding protein as desired. Alternatively, variants can be designed to alter the biological activity of the antigen binding protein. For example, glycosylation sites can be altered or removed.

[1020] Various antibodies can be used together with the ADC used herein to target cancer cells. As shown below, the linker loading in the ADC disclosed herein is unexpectedly effective for different tumor antigen targeting antibodies. Suitable antigens expressed on cancer cells but not on healthy cells, or suitable antigens expressed on cancer cells at a level higher than that on healthy cells are known in the art, as are antibodies against them. Those skilled in the art can prepare other antibodies against those antigen targets. These antibodies can be used together with the linker and Bcl-xL inhibitor loading disclosed herein. In some embodiments, antibodies or antigen-binding fragments targeting EphA2 provide specific improved drugs: antibody ratio, aggregation level, stability (i.e., in vitro and in vivo stability), tumor targeting (i.e., cytotoxicity, efficacy), minimization of off-target killing and / or therapeutic efficacy. The improved therapeutic efficacy can be measured in vitro or in vivo and may include a slowed tumor growth rate and / or a reduced tumor volume.

[1021] In some embodiments, alternative antibodies directed against the same target or antibodies directed against different antigenic targets are used and provide at least some of the advantageous functional properties described above (e.g., improved stability, improved tumor targeting, improved therapeutic efficacy, etc.).

[1022] connector

[1023] In some embodiments, the linker in the ADC is extracellularly stable in a manner sufficient to be therapeutically effective. In some embodiments, the linker is stable outside the cell so that the ADC remains intact when present in extracellular conditions (e.g., prior to transport or delivery into the 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., a Bcl-xL inhibitor).

[1024] As used herein, "stable" in the context of a linker or an ADC comprising a linker means that when the ADC is present under extracellular conditions, no more than 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% of the linker (or any percentage therebetween) in the ADC sample is cleaved (or in the case of an incomplete ADC as a whole). In some embodiments, the linker and / or ADC disclosed herein is stable compared to alternative linkers and / or ADCs with alternative linkers and / or Bcl-xL inhibitor loads. In some embodiments, the ADC disclosed herein can 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.

[1025] Whether the linker is stable outside the cell can be determined, for example, by including 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 can give the ADC time to localize to the targeted cancer cells and prevent premature release of the drug moiety, which may reduce the therapeutic index of the ADC by indiscriminately damaging normal tissue and cancer tissue. In some embodiments, the linker is stable outside the target cell and releases the drug moiety from the ADC once inside the cell so that the drug can bind to its target. Thus, an effective linker will: (i) maintain the specific binding properties of the antibody or antigen-binding fragment; (ii) allow delivery (e.g., intracellular delivery) of the drug moiety via stable attachment to the antibody or antigen-binding fragment; (iii) remain stable and intact until the ADC is transported or delivered to its target site; and (iv) allow the therapeutic effect of the drug moiety after cleavage or alternative release mechanism, e.g., cytotoxic effect.

[1026] Linkers may affect the physicochemical properties of ADCs. Since many cytotoxic agents are hydrophobic in nature, linking them to antibodies with additional hydrophobic moieties may result in aggregation. ADC aggregates are insoluble and often limit the drug loading achievable on the antibody, which can negatively impact the efficacy of the ADC. In general, protein aggregates of biologics are also associated with increased immunogenicity. As shown below, the linkers disclosed herein produce ADCs with low aggregation levels and desired drug loading levels.

[1027] 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., a Bcl-xL inhibitor) upon exposure to certain environmental cues (e.g., upon internalization into target cells), while non-cleavable linkers typically rely on degradation of the antibody or antigen-binding fragment itself.

[1028] As used herein, term " alkyl " refers to the straight chain or branched hydrocarbon chain group being only composed of carbon and hydrogen atom, and there is no unsaturation in described group.As used herein, term " C1-C6 alkyl " refers to the straight chain or branched hydrocarbon chain group being only composed of carbon and hydrogen atom, and there is no unsaturation in described group, there is from one to six carbon atom, and is attached to the remainder of molecule by singly-bound.The limiting examples of " C1-C6 alkyl " group includes methyl (C1 alkyl), ethyl (C2 alkyl), 1- methylethyl (C3 alkyl), n-propyl (C3 alkyl), isopropyl (C3 alkyl), normal-butyl (C4 alkyl), isobutyl (C4 alkyl), sec-butyl (C4 alkyl), the tert-butyl (C4 alkyl), n-pentyl (C5 alkyl), isopentyl (C5 alkyl), neopentyl (C5 alkyl) and hexyl (C6 alkyl).

[1029] As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, said radical containing at least one double bond. As used herein, the term "C2-C6 alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, said radical containing at least one double bond, having from two to six carbon atoms, attached to the remainder of the molecule by a single bond. Non-limiting examples of "C2-C6 alkenyl" groups include vinyl (C2 alkenyl), prop-1-enyl (C3 alkenyl), but-1-enyl (C4 alkenyl), pent-1-yl (C5 alkenyl), pent-4-yl (C5 alkenyl), pent-1,4-dienyl (C5 alkenyl), hexa-1-enyl (C6 alkenyl), hexa-2-enyl (C6 alkenyl), hexa-3-enyl (C6 alkenyl), hexa-1-,4-dienyl (C6 alkenyl), hexa-1-,5-dienyl (C6 alkenyl) and hexa-2-,4-dienyl (C6 alkenyl). As used herein, the term "C2-C3 alkenyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing at least one double bond, having from two to three carbon atoms, attached to the remainder of the molecule by a single bond. Non-limiting examples of "C2-C3 alkenyl" groups include vinyl (C2 alkenyl) and prop-1-enyl (C3 alkenyl).

[1030] As used herein, term " alkylidene " refers to the divalent straight chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, and there is no unsaturation in the group. As used herein, term " C1-C6 alkylidene " refers to the divalent straight chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, and there is no unsaturation in the group, with from one to six carbon atoms. The limiting examples of " C1-C6 alkylidene " group include methylene (C1 alkylidene), ethylene (C2 alkylidene), 1- methylethylene (C3 alkylidene), n-propylene (C3 alkylidene), isopropylidene (C3 alkylidene), n-butylidene (C4 alkylidene), isobutylidene (C4 alkylidene), secondary butylidene (C4 alkylidene), tert-butylidene (C4 alkylidene), n-pentylene (C5 alkylidene), isopentylene (C5 alkylidene), neopentylene (C5 alkylidene) and hexylene (C6 alkylidene).

[1031] As used herein, the term "alkenylene" refers to a divalent straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, said group containing at least one double bond. As used herein, the term "C2-C6 alkenylene" refers to a divalent straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, said group containing at least one double bond, and having from two to six carbon atoms. Non-limiting examples of "C2-C6 alkenylene" groups include vinylene (C2 alkenylene), prop-1-alkenylene (C3 alkenylene), but-1-alkenylene (C4 alkenylene), pent-1-alkenylene (C5 alkenylene), pent-4-alkenylene (C5 alkenylene), pent-1,4-dialkenylene (C5 alkenylene), hex-1-alkenylene (C6 alkenylene), hex-2-alkenylene (C6 alkenylene), hex-3-alkenylene (C6 alkenylene), hex-1-,4-dialkenylene (C6 alkenylene), hex-1-,5-dialkenylene (C6 alkenylene) and hex-2-,4-dialkenylene (C6 alkenylene). As used herein, the term "C2-C6 alkenylene" 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 two to three carbon atoms. Non-limiting examples of "C2-C3 alkenylene" groups include ethenylene (C2 alkenylene) and prop-1-enylene (C3 alkenylene).

[1032] As used herein, the term "cycloalkyl" or "C3-C8 cycloalkyl" 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 adamantyl. Non-limiting examples of monocyclic C3-C8 cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[1033] The term "aryl" as used herein refers to phenyl, naphthyl, biphenyl or indenyl.

[1034] The term "heteroaryl" as used herein refers to any monocyclic or bicyclic group consisting of 5 to 10 ring members having at least one aromatic moiety and containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen (including quaternary nitrogen).

[1035] The term "cycloalkyl" as used herein refers to any monocyclic or bicyclic non-aromatic carbocyclic group containing 3 to 10 ring members, which may include fused, bridged, or spirocyclic systems. Non-limiting examples of fused bicyclic or bridged 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, and bicyclo[2.2.2]octane. Non-limiting examples of monocyclic C3-C8 cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[1036] The term "heterocycloalkyl" refers to any monocyclic or bicyclic non-aromatic carbocyclic group consisting of 3 to 10 ring members and containing 1 to 3 heteroatoms selected from oxygen, sulfur, SO, SO2 and nitrogen, it being understood that the bicyclic group may be of the fused or spiro type. C3-C8 heterocycloalkyl refers to a heterocycloalkyl group having 3 to 8 ring carbon atoms. A heterocycloalkyl group may have 4 to 10 ring members.

[1037] The terms heteroarylene, cycloalkylene, heterocycloalkylene refer to divalent heteroaryl, cycloalkyl and heterocycloalkyl groups.

[1038] The term "haloalkyl" as used herein refers to a straight or branched alkyl chain that is substituted with one or more halogen groups in place of hydrogen along the hydrocarbon chain. Examples of halogen groups that are suitable for substitution in haloalkyl include fluorine, bromine, chlorine, and iodine. Haloalkyl can include substituting hydrogen in the alkyl chain with multiple halogen groups, wherein the halogen groups can be attached to the same carbon or another carbon in the alkyl chain.

[1039] As used herein, alkyl, alkenyl, alkynyl, alkoxy, amino, aryl, heteroaryl, cycloalkyl, and heterocycloalkyl groups may be optionally substituted with 1 to 4 groups selected from the group consisting of optionally substituted straight or branched (C1-C6) alkyl, optionally substituted straight or branched (C2-C6) alkenyl, optionally substituted straight or branched (C2-C6) alkynyl, optionally substituted straight or branched (C1-C6) alkoxy, optionally substituted (C1-C6) alkyl-S-, hydroxy, oxo (or N-oxo where appropriate), compounds), nitro, cyano, -C(O)-OR0', -OC(O)-R0', -C(O)-NR0'R0", -NR0'R0", -(C=NR0')-OR0", linear or branched (C1-C6)haloalkyl, trifluoromethoxy, or halogen, wherein R0' and R0" are each independently a hydrogen atom or an optionally substituted linear or branched (C1-C6)alkyl, and wherein one or more carbon atoms of the linear or branched (C1-C6)alkyl are optionally deuterated.

[1040] As used herein, the term "polyoxyethylene," "polyethylene glycol," or "PEG" refers to a linear, branched, or star-shaped configuration consisting of (OCH2CH2) groups. In certain embodiments, the polyethylene or PEG group is -(OCH2CH2) t *-, wherein t is 1-40 or 4-40, and wherein "-" indicates the end pointing to the self-immolative spacer and "*-" indicates the point of attachment to the terminal group R', wherein R' is OH, OCH3 or OCH2CH2C(=O)OH. In other embodiments, the polyethylene or PEG group is -(CH2CH2O) t *-, wherein t is 1-40 or 4-40, wherein "-" indicates pointing to the end of the self-immolative spacer and "*-" indicates the point of attachment to the terminal group R", wherein R" is H, CH3 or CH2CH2C(=O)OH. For example, the term "PEG12" as used herein means that t is 12.

[1041] The term "polyalkylene glycol" as used herein refers to a glycol consisting of (O(CH2) m ) n In certain embodiments, the polyethylene or PEG group is -(O(CH2) m ) t *-, wherein m is 1-10, t is 1-40 or 4-40, and wherein "-" indicates the end pointing to the self-immolative spacer and "*-" indicates the point of attachment to the terminal group R', wherein R' is OH, OCH3 or OCH2CH2C(=O)OH. In other embodiments, the polyethylene or PEG group is -((CH2) m O) t*-, wherein m is 1-10, t is 1-40 or 4-40, and wherein "-" indicates pointing to the end of the self-immolative spacer and "*-" indicates the point of attachment to the terminal group R', wherein R' is H, CH3 or CH2CH2C(=O)OH.

[1042] As used herein, the term "reactive group" is a functional group that is capable of forming a covalent bond with a functional group of an antibody, an antibody fragment, or another reactive group attached to an antibody or an antibody fragment. Non-limiting examples of such functional groups include the reactive groups of Table 3 provided herein.

[1043] As used herein, the term "attachment group" or "coupling group" refers to a divalent moiety that connects a bridging spacer to an antibody or fragment thereof. An attachment or coupling group is a divalent moiety formed by reaction between a reactive group and a functional group on the antibody or fragment thereof. Non-limiting examples of such divalent moieties include the divalent chemical moieties given in Tables 3 and 4 provided herein.

[1044] As used herein, the term "bridging spacer" refers to one or more linker components covalently attached together to form a bivalent moiety that connects a bivalent peptide spacer to a reactive group, connects a bivalent peptide spacer to a coupling group, or connects an attachment group to at least one cleavable group. In certain embodiments, the "bridging spacer" comprises a carboxyl group attached to the N-terminus of the bivalent peptide spacer via an amide bond.

[1045] As used herein, the term "spacer moiety" refers to one or more linker components that are covalently attached together to form a moiety that connects the self-immolative spacer to the hydrophilic moiety.

[1046] As used herein, the term "divalent peptide spacer" refers to a divalent linker comprising one or more amino acid residues covalently attached together to form a portion connecting the bridging spacer to the self-immolative spacer. The one or more amino acid residues can be 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), valeric acid (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine ​​and demethylpyrrolysine.

[1047] In certain embodiments, the "divalent peptide spacer" is a combination of 2 to 4 amino acid residues, wherein each residue is independently 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), (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), valeric acid (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine ​​and demethylpyrrolysine, such as -ValCit*; -CitVal*; -AlaAla*; -AlaCit*; -CitAla*; -AsnCit*; -C itAsn*;-CitCit*;-ValGlu*;-GluVal*;-SerCit*;-CitSer*;-LysCit*;-CitLys*;-AspCit*;-CitAsp*;-A laVal*;-ValAla*;-PheAla*;-AlaPhe*;-PheLys*;-LysPhe*;-ValLys*;-LysVal*;-AlaLys*;-LysAla*;-P heCit*;-CitPhe*;-LeuCit*;-CitLeu*;-IleCit*;-CitIle*;-PheArg*;-ArgPhe*;-CitTrp*;-TrpCit*;-P hePheLys*; -LysPhePhe*; -DPhePheLys*; -DLysPhePhe*; -GlyPheLys*; -LysPheGly*; -GlyPheLeuGly-[SEQ ID NO:62]; -GlyLeuPheGly-[SEQ ID NO:63]; -AlaLeuAlaLeu-[SEQ ID NO:64], -GlyGlyGly*; -GlyGlyGlyGly-[SEQ ID NO:65]; -GlyPheValGly-[SEQ ID NO:66]; and -GlyValPheGly-[SEQ ID NO:67], wherein "-" indicates the point of attachment to the bridging spacer and "*" indicates the point of attachment to the self-immolative spacer.

[1048] As used herein, the term "linker component" refers to a chemical moiety that is part of a linker. Examples of linker components include: alkylene groups: -(CH2)n -, which may be linear or branched (in which case n is 1-18); alkenylene group; alkynylene group; alkenyl group; alkynyl group; ethylene glycol unit: -OCH2CH2- or -CH2CH2O-; polyethylene glycol unit: (-CH2CH2O-) x (wherein x is 2-20 in this example); -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; sulfonamides: -S(O)2NH- or -NHS(O)2; ether: -CH2O- or -OCH2-; alkylene substituted by one or more groups independently selected from carboxyl, sulfonate, hydroxyl, amine, amino acid, sugar, phosphoric acid and phosphonate; alkenylene substituted by one or more groups independently selected from carboxyl, sulfonate, hydroxyl, amine, amino acid, sugar, phosphoric acid and phosphonate; alkynylene substituted by one or more groups independently selected from carboxyl, sulfonate, hydroxyl, amine, amino acid, sugar, phosphoric acid and phosphonate; C1-C 10Alkylene, wherein one or more methylene groups are replaced by one or more -S-, -NH-, or -O- moieties; a ring system having two available attachment points, for example, selected from 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; an amino acid residue 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 (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), valeric acid (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine ​​and demethylpyrrolysine; a combination of two or more amino acid residues, each of which is independently 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 (Arg), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), citrulline (Cit), valeric acid (Nva), norleucine (Nle), selenocysteine ​​(Sec), pyrrolysine (Pyl), homoserine, homocysteine ​​and demethylpyrrolysine, such as Val-Cit; Cit-Val; Ala-Ala; A la-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-L ys; 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 a self-immolative spacer, wherein the self-immolative spacer comprises one or more protecting (triggering) groups that are sensitive 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.

[1049] Non-limiting examples of such self-immolative spacers include:

[1050] in:

[1051] PG is a protecting (triggering) group;

[1052] X a is O, NH or S;

[1053] X b is O, NH, NCH3 or S;

[1054] X c is O or NH;

[1055] Y a is CH2, CH2O or CH2NH;

[1056] Y b is CH2, O or NH;

[1057] Y c is a bond, CH2, O, or NH, and

[1058] LG is a leaving group, such as the drug portion (D) of a linker-drug group of the present invention.

[1059] Additional non-limiting examples of such self-immolative spacers are described in Angew. Chem. Int. Ed. 2015, 54, 7492-7509.

[1060] Additionally, the 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 given in Table 3.

[1061] Table 3

[1062]

[1063]

[1064]

[1065]

[1066]

[1067] Where: R in Table 3 32 H, C 1-4 Alkyl, phenyl, pyrimidine or pyridine; R in Table 3 35 H, C 1-6 Alkyl, phenyl or C substituted by 1 to 3 -OH groups 1-4 Alkyl; each R in Table 3 7 Independently selected from H, C 1-6 Alkyl, fluorine, benzyloxy substituted by -C(=O)OH, benzyl substituted by -C(=O)OH, C 1-4 Alkoxy and C substituted by -C(=O)OH 1-4 Alkyl; R in Table 3 37 are independently selected from H, phenyl and pyridine; q in Table 3 is 0, 1, 2 or 3; R in Table 3 8 and R 13 is H or methyl; R in Table 3 9 and R 14 is H, -CH3 or phenyl; R in Table 3 is H or any suitable substituent; and

[1068] R in Table 3 50 For H.

[1069] Additionally, the linker component may be a group listed in Table 4 below.

[1070] Table 4.

[1071]

[1072]

[1073]

[1074] As used herein, when showing a partial structure of a compound, a wavy line Indicates the point of attachment of a partial structure to the rest of the molecule.

[1075] As used herein, the terms "self-immolative spacer" and "self-immolative group" refer to a moiety comprising one or more trigger groups (TGs) that are activated by 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, and upon activation, the protecting group is removed, which results in a cascade of decomposition reactions, leading to the release of the leaving group in a time sequence. This cascade reaction can be, but is not limited to, a 1,4-, 1,6-, or 1,8-elimination reaction.

[1076] Non-limiting examples of self-immolative spacers or groups include:

[1077] wherein these groups may be optionally substituted, and

[1078] in:

[1079] TG is the trigger group;

[1080] X a is O, NH or S;

[1081] X b is O, NH, NCH3 or S;

[1082] X c is O or NH;

[1083] Y a is CH2, CH2O or CH2NH;

[1084] Y b is CH2, O or NH;

[1085] Y c is a bond, CH2, O, or NH, and

[1086] LG is a leaving group, such as the drug portion (D) of a linker-drug group of the present invention.

[1087] Additional non-limiting examples of self-immolative spacers are described in Angew. Chem. Int. Ed. 2015, 54, 7492-7509.

[1088] In certain embodiments, the self-immolative spacer is a moiety having the structure

[1089] wherein Lp is an enzymatically cleavable divalent peptide spacer and A, D, L3 and R 2 As defined herein.

[1090] In a preferred embodiment, the self-immolative spacer is a moiety having the structure

[1091] Where Lp is an enzymatically cleavable divalent peptide spacer and D, L3 and R 2 As defined herein. In some embodiments, D is a Bcl-xL inhibitor containing a quaternized tertiary amine.

[1092] In other preferred embodiments, the self-immolative spacer is a moiety having the structure

[1093] Where Lp is an enzymatically cleavable divalent peptide spacer and D, L3 and R 2 As defined herein.

[1094] As used herein, the term "hydrophilic moiety" refers to a moiety having a hydrophilic nature that increases the water solubility of the drug moiety (D) when attached to the linker group of the present invention. Examples of such hydrophilic groups include, but are not limited to, polyethylene glycol, polyalkylene glycol, sugars, oligosaccharides, A polypeptide containing a C2-C6 alkyl group substituted with a C2-C6 alkyl group.

[1095] Drug section

[1096] In some embodiments, the intermediate as a precursor of the linker moiety reacts with the drug moiety (e.g., a Bcl-xL inhibitor) under appropriate conditions. In some embodiments, a reactive group is used on the drug and / or intermediate or linker. The reaction product between the drug and the intermediate or the derivatized drug (drug plus linker) is then reacted with the antibody or antigen-binding fragment under conditions that are conducive to the conjugation of the drug and the intermediate or the derivatized drug and the antibody or antigen-binding fragment. Alternatively, the intermediate or linker can first react with the antibody or antigen-binding fragment, or the derivatized antibody or antigen-binding fragment, and then react with the drug or the derivatized drug.

[1097] Many different reactions can be used to covalently attach the drug moiety and / or linker moiety to the antibody or antigen-binding fragment. This is typically accomplished by reacting one or more amino acid residues of the antibody or antigen-binding fragment, including the amine group of lysine, the free carboxylic acid groups of glutamic acid and aspartic acid, the sulfhydryl group of cysteine, and different portions of aromatic amino acids. For example, a carbodiimide reaction can be used for nonspecific covalent attachment to connect the carboxyl (or amino) group on the drug moiety to the amino (or carboxyl) group on the antibody or antigen-binding fragment. In addition, bifunctional reagents such as dialdehydes or imidoesters can also be used to connect the amino group on the drug moiety to the amino group on the antibody or antigen-binding fragment. The Schiff base reaction can also be used to attach drugs (e.g., Bcl-xL inhibitors) to binding agents. This method involves periodate oxidation of drugs containing diols or hydroxyls to form aldehydes, which are then reacted with the binding agent. Attachment occurs by forming a Schiff base with the amino group of the binding agent. Isothiocyanates can also be used as coupling agents for covalently attaching 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 produced and attached to antibodies or antigen-binding fragments using various chemistries known in the art include Bcl-xL inhibitors, such as those described and exemplified herein.

[1098] Suitable drug moieties may comprise a compound of Formula (I), (IA), (IB), (IC), (II), (IIA), (IIB), or (IIC), or an enantiomer, diastereomer, and / or addition salt thereof with a pharmaceutically acceptable acid or base. Additionally, the drug moiety may comprise any of the compounds described herein as Bcl-xL inhibitors (D).

[1099] In some embodiments, the drug moiety (D) comprises a formula selected from Table A2.

[1100] In some embodiments, the drug moiety (D) comprises a Bcl-xL inhibitor known in the art, such as ABT-737 and ABT-263.

[1101] In some embodiments, the drug moiety (D) comprises a Bcl-xL inhibitor selected from:

[1102]

[1103]

[1104] In some embodiments, the Linker-Drug (or "Linker-Load") Moiety-(LD) may comprise a compound in Table B, or an enantiomer, diastereomer, deuterated derivative, and / or pharmaceutically acceptable salt of any of the foregoing.

[1105] Drug loading

[1106] Drug load is represented by p, and is also referred to herein as drug-to-antibody ratio (DAR). Drug load can be 1 to 16 drug moieties / 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.

[1107] Drug load may be subject to the limitation of the number of attachment sites on antibody or Fab.In some embodiments, the linker portion (L) of ADC is attached to antibody or Fab by the chemically active group on one or more amino acid residues on antibody or Fab.For example, the linker can be attached to antibody or Fab by free amino, imino, hydroxyl, thiol or carboxyl group (for example, N- or C-terminal, the ε amino group of one or more lysine residues, the free carboxylic acid group of one or more glutamic acid or aspartic acid residues or the sulfhydryl group of one or more cysteine ​​residues).The site of linker attachment can be the natural residue in the amino acid sequence of antibody or Fab, or it can be introduced into antibody or Fab, for example, by DNA recombination technology (for example, by introducing cysteine ​​residues into amino acid sequence) or by protein biochemistry (for example, by reduction, pH adjustment or hydrolysis).

[1108] In some embodiments, the number of drug moieties that can be conjugated to an antibody or Fab is limited by the number of free cysteine ​​residues. For example, when attached to a cysteine ​​thiol group, the antibody may have only one or several cysteine ​​thiol groups, or may have only one or several linkers that can be attached via a thiol group with sufficient reactivity. Typically, antibodies do not contain many free and reactive cysteine ​​thiol groups that can be connected to the drug moiety. In fact, most of the cysteine ​​thiol residues in antibodies are related to interchain or intrachain disulfide bonds. Therefore, in some embodiments, conjugation with cysteine ​​may require at least partial reduction of the antibody. Excessive attachment of the linker-toxin to the antibody can destabilize the antibody by reducing the cysteine ​​residues that can be used to form a disulfide bond. Therefore, the best drug: antibody ratio should increase the effectiveness of the ADC (by increasing the number of drug moieties attached to each antibody) without destabilizing the antibody or Fab. In some embodiments, the best ratio can be 2, 4, 6, or 8. In some embodiments, the best ratio can be 2 or 4.

[1109] In some embodiments, the antibody or antigen-binding fragment is exposed to reducing conditions to produce one or more free cysteine ​​residues before conjugation. In some embodiments, the antibody can be reduced with a reducing agent such as dithiothreitol (DTT) or tris (2-carboxyethyl) phosphine (TCEP) under partial or total reducing conditions to generate reactive cysteine ​​thiol groups. Unpaired cysteines can be produced by partial reduction with a limited molar equivalent of TCEP, which can reduce the interchain disulfide bonds connecting the light chain and the heavy chain (each HL is paired with one pair) and the two heavy chains in the hinge region (each HH is paired with two pairs in the case of human IgG1), while keeping the intrachain disulfide bonds intact (Stefano et al. (2013) Methods Mol Biol. 1045: 145-71). In embodiments, the disulfide bonds in the antibody are electrochemically reduced, for example, by using a working electrode that applies alternating reduction and oxidation voltages. This method can allow disulfide bond reduction to be coupled online with 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 chromatograph (e.g., HPLC) or an electrophoresis device (see, e.g., US 2014 / 0069822). In some embodiments, the antibody is subjected to denaturing conditions to reveal reactive nucleophilic groups on amino acid residues (e.g., cysteine).

[1110] Drug loading of ADCs can be controlled in various ways, such as: (i) limiting the molar excess of drug-linker intermediates or linker reagents relative to the antibody; (ii) limiting the conjugation reaction time or temperature; (iii) partial or limited reducing conditions for cysteine ​​thiol modification; and / or (iv) engineering the amino acid sequence of the antibody by recombinant techniques to modify the number and position of cysteine ​​residues to control the number and / or position of linker-drug attachments.

[1111] In some embodiments, free cysteine ​​residues are introduced into the amino acid sequence of an antibody or antigen-binding fragment. For example, a cysteine ​​engineered antibody can be prepared in which one or more amino acids of a parent antibody are replaced by cysteine ​​amino acids. Any form of antibody can be engineered in this way, i.e., mutated. For example, a parent Fab antibody fragment can be engineered to form a cysteine ​​engineered Fab referred to as a "ThioFab". Similarly, a parent monoclonal antibody can be engineered to form a "ThioMab". Single-site mutations produce a single engineered cysteine ​​residue in a ThioFab, while due to the dimeric nature of IgG antibodies, single-site mutations produce two engineered cysteine ​​residues in a ThioMab. DNA encoding amino acid sequence variants of a parent polypeptide can be prepared by various methods known in the art (see, for example, the method 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 a DNA encoding a polypeptide prepared earlier. Variants of recombinant antibodies can also be constructed by restriction fragment manipulation or by overlap extension PCR using synthetic oligonucleotides. ADCs of formula (1) include, but are not limited to, antibodies with 1, 2, 3, or 4 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 the antibody or Fab, and no engineering is required. In this case, the free cysteine ​​residues present can be used to couple the antibody or Fab to the drug moiety.

[1112] In a reaction mixture comprising multiple copies of an antibody or antigen-binding fragment and a linker moiety, when more than one nucleophilic group reacts with a drug-linker intermediate or a linker moiety reagent and then reacts with a drug moiety reagent, the resulting product may be a mixture of ADC compounds in which one or more drug moieties of each copy of the antibody or antigen-binding fragment attached to the mixture are distributed. In some embodiments, the drug loading in the ADC mixture obtained by the conjugation reaction is in the range of 1 to 16 attached drug moieties / antibody or antigen-binding fragment. The average number of drug moieties / antibody or antigen-binding fragment (i.e., average drug loading 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 / 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 from about 1.5 to about 3.5, from about 2.5 to about 4.5, from about 3.5 to about 5.5, from about 4.5 to about 6.5, from about 5.5 to about 7.5, from about 6.5 to about 8.5, or from about 7.5 to about 9.5. In some embodiments, the average number of drug moieties per antibody or antigen-binding fragment is from about 2 to about 4, from about 3 to about 5, from about 4 to about 6, from about 5 to about 7, from about 6 to about 8, from about 7 to about 9, from about 2 to about 8, or from about 4 to about 8.

[1113] 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.

[1114] 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.

[1115] In some embodiments, when used with respect to the average number of drug moieties per antibody or antigen-binding fragment, the term "about" means plus or minus 20%, 15%, 10%, 5%, or 1%. In one embodiment, the term "about" refers to a range of values ​​that is 10% more or less than a specified value. In another embodiment, the term "about" refers to a range of values ​​that is 5% more or less than a specified value. In another embodiment, the term "about" refers to a range of values ​​that is 1% more or less than a specified value.

[1116] Individual ADC complexes or "species" can be identified in the mixture by mass spectrometry and separated by, for example, UPLC or HPLC (e.g., hydrophobic interaction chromatography (HIC-HPLC). In some embodiments, a homogeneous or near-homogeneous ADC product having a single loading value can be separated from the conjugated mixture, for example, by electrophoresis or chromatography.

[1117] In some embodiments, higher drug loading (e.g., p>16) may result in aggregation, insolubility, toxicity, or loss of cell permeability of certain antibody-drug conjugates. Higher drug loading may also have a negative impact on the pharmacokinetics (e.g., clearance) of certain ADCs. In some embodiments, lower drug loading (e.g., p<2) can reduce the effectiveness of certain ADCs for target-expressing cells. In some embodiments, the drug loading range of the ADCs of the present disclosure is from about 2 to about 16, from about 2 to about 10, from about 2 to about 8; from about 2 to about 6; from about 2 to about 5; from about 3 to about 5; from about 2 to about 4; or from about 4 to about 8.

[1118] In some embodiments, for example, partial reduction of intrachain disulfide bonds on an antibody or antigen-binding fragment is used to achieve a drug load and / or average drug load of about 2, and to provide beneficial properties. In some embodiments, for example, partial reduction of intrachain disulfide bonds on an antibody or antigen-binding fragment is used to achieve a drug load and / or average drug load of about 4, about 6, or about 8, and to provide beneficial properties. In some embodiments, a drug load and / or average drug load of less than about 2 may result in unacceptably high levels of unconjugated antibody species, which may compete with the ADC for binding to the target antigen and / or provide reduced therapeutic efficacy. In some embodiments, a drug load and / or average drug load of more than about 16 may result in unacceptably high levels 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 the loss of one or more chemical bonds required to stabilize the antibody or antigen-binding fragment.

[1119] The present disclosure includes methods for producing the described ADC. In short, the ADC comprises an antibody or antigen-binding fragment as an antibody or antigen-binding fragment, a drug moiety (e.g., a Bcl-xL inhibitor) and a linker connecting the drug moiety and the antibody or antigen-binding fragment. In some embodiments, a linker having a reactive functional group for covalent attachment to the drug moiety and the antibody or antigen-binding fragment can be used to prepare the ADC. In some embodiments, the antibody or antigen-binding fragment is functionalized to prepare a functional group that reacts with a linker or a drug-linker intermediate. For example, in some embodiments, the cysteine ​​thiol of the antibody or antigen-binding fragment can form a key with a reactive functional group of a linker or a drug-linker intermediate to prepare the ADC. In some embodiments, the antibody or antigen-binding fragment is prepared using a bacterial transglutaminase (BTG)-reactive glutamine that is specifically functionalized with an amine containing a cyclooctene BCN (N-[(1R,8S,9s)-bicyclo[6.1.0]non-4-yn-9-ylmethyloxycarbonyl]-1,8-diamino-3,6-dioxooctane) 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. The production of ADCs can be accomplished by techniques known to those skilled in the art.

[1120] In some embodiments, an ADC is produced by contacting an antibody or antigen-binding fragment with a linker and a drug moiety (e.g., a Bcl-xL inhibitor) in a sequential manner, such that the antibody or antigen-binding fragment is first covalently linked to the linker and then the 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, an ADC is produced by contacting an antibody or antigen-binding fragment with a linker-drug compound preformed by reacting a linker with a 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 the drug moiety in a reaction mixture, allowing covalent bonds to form simultaneously between the antibody or antigen-binding fragment and the linker and between the linker and the drug moiety. The method for producing an ADC may include a reaction in which the antibody or antigen-binding fragment is contacted with the antibody or antigen-binding fragment before the linker is added to the reaction mixture, or vice versa. In some embodiments, an ADC is produced by reacting an antibody or antigen-binding fragment with a linker conjugated to a drug moiety (e.g., a Bcl-xL inhibitor) under conditions that allow conjugation.

[1121] The ADC prepared according to the above method can be subjected to a purification step. The purification step may involve any biochemical method known in the art for purifying proteins or any combination thereof. These include, but are not limited to, tangential flow filtration (TFF), affinity chromatography, ion exchange chromatography, chromatography based on any charge or isoelectric point, mixed mode chromatography, such as CHT (ceramic hydroxyapatite), hydrophobic interaction chromatography, size exclusion chromatography, dialysis, filtration, selective precipitation, or any combination thereof.

[1122] Therapeutic uses and compositions

[1123] Disclosed herein are methods for treating a subject's disorder (e.g., cancer) using compositions described herein (e.g., disclosed ADC compounds and compositions). Compositions, such as ADCs, can be administered alone or in combination with at least one additional inactive agent and / or active agent, such as at least one additional therapeutic agent, and can be administered in any pharmaceutically acceptable formulation, dosage, and dosing regimen. The toxicity and efficacy indicators of the therapeutic effect can be assessed and adjusted accordingly. Efficacy measurements include, but are not limited to, cell inhibition and / or cytotoxic effects observed in vitro or in vivo, tumor volume reduction, tumor growth inhibition, and / or prolonged survival.

[1124] Methods for determining whether an ADC exerts a cytostatic and / or cytotoxic effect on a cell are known. For example, the cytotoxic or cytostatic activity of an ADC can be measured by, for example, exposing mammalian cells expressing the ADC target antigen to a cell culture medium; culturing the cells for a period of about 6 hours to about 6 days; and measuring cell viability (e.g., using a CellTiter- (CTG or MTT cell viability assay). Cell-based in vitro assays can also be used to measure viability (proliferation), cytotoxicity, and apoptosis induction (caspase activation) of ADCs.

[1125] In order to determine cytotoxicity, necrosis or apoptosis (programmed cell death) can be measured. Necrosis is usually accompanied by increased plasma membrane permeability, cell swelling and plasma membrane rupture. For example, apoptosis can be quantitatively measured by measuring DNA fragmentation. Commercial photometry can be used for quantitative in vitro DNA fragmentation determination. The example of such determination method, including TUNEL (it detects the incorporation of labeled nucleotides in fragmented DNA) and ELISA-based determination method, is described in Biochemica (1999) 2:34-7 (Roche Molecular Biochemicals).

[1126] Apoptosis can also be determined by measuring morphological changes in the cells. For example, as with necrosis, loss of plasma membrane integrity can be determined by measuring the uptake of certain dyes (e.g., fluorescent dyes such as acridine orange or ethidium bromide). One method for measuring the number of apoptotic cells is described in Duke and Cohen, Current Protocols in Immunology (Coligan et al., eds. (1992) pp. 3.17.1-3.17.16). Cells can also be labeled with DNA dyes (e.g., acridine orange, ethidium bromide, or propidium iodide) and observing the chromatin condensation and margination along the inner nuclear membrane of the cells. In some embodiments, apoptosis can also be determined by screening for caspase activity. In some embodiments, caspase- The assay can 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 reagents optimized for caspase activity, luciferase activity, and cell lysis. In some embodiments, caspase-3 / 7 substrates are added in an "add-mix-measure" format. The 3 / 7 reagent may cause cell lysis, followed by caspase cleavage of the substrate and generation of a "luminescent" luminescent signal produced by luciferase. In some embodiments, luminescence may be proportional to the amount of caspase activity present and may 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 may be useful in treating cancer.

[1127] This can be achieved, for example, by measuring the expression of dyes such as neutral red, trypan blue, crystal violet or ALAMAR in cells. TM Cell viability is measured by the uptake of dapoxetine blue (see, e.g., Page et al. (1993) Intl J Oncology 3:473-6). In such an assay, cells are incubated in medium containing the dye, the cells are washed, and the remaining dye, which reflects the uptake of the dye by the cells, is measured spectrophotometrically.

[1128] Cell viability can also be measured, for example, by quantifying ATP (an indicator of metabolically active cells). In some embodiments, the CellTiter- (CTG) cell viability to evaluate the in vitro potency and / or cell viability assays of prepared ADC or Bcl-xL inhibitor compounds, as described in the embodiments provided herein. In this assay, in some embodiments, a single reagent (CellTiter- The 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 proportional to the number of cells in the culture.

[1129] Cell viability can also be measured, for example, by measuring the reduction of tetrazolium salts. In some embodiments, MTT cell viability can be used to assess the in vitro efficacy and / or cell viability assay of the prepared ADC or Bcl-xL inhibitor compound, as described in the embodiments provided herein. In this assay, in some embodiments, yellow tetrazolium MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) is partially reduced by the action of dehydrogenases by metabolically active cells to produce equivalents that reduce NADH and NADPH, etc. The resulting intracellular purple formazan can then be dissolved and quantified by spectrophotometry.

[1130] In certain aspects, the present disclosure features a method for killing, inhibiting, or regulating cancer cell or tissue growth by disrupting the expression and / or activity of Bcl-xL and / or one or more of its upstream regulators or its downstream targets. The method can be used for any subject in which disruption of Bcl-xL expression and / or activity provides a therapeutic benefit. Subjects who may benefit from disruption of Bcl-xL expression and / or activity include, but are not limited to, subjects with or at risk of developing a cancer, such as a tumor or hematological cancer. In some embodiments, the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, gastric cancer, bladder cancer, brain cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colorectal cancer, esophageal cancer, hepatocellular carcinoma, lymphoblastic leukemia (including acute lymphocytic 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, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer. In some embodiments, the cancer is lymphoma or gastric cancer.

[1131] In some embodiments, the disclosed ADCs can be administered to any cell or tissue that expresses EphA2, such as a cancer cell or tissue that expresses EphA2. Exemplary embodiments include methods for killing cancer cells or tissue that express EphA2. The methods can be used for any cell or tissue that expresses EphA2, such as a cancer cell or metastatic lesion. Non-limiting examples of cancers that express EphA2 include breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer, and colon cancer. Non-limiting examples of cells that express EphA2 include EBC-1 cells and cells containing a recombinant nucleic acid encoding EphA2 or a portion thereof.

[1132] Exemplary methods include the step of contacting cells with an effective amount (i.e., an amount sufficient to kill the cells) of an ADC, as described herein. The method can be used with cultured cells, for example, in vitro, in vivo, ex vivo, or in situ. For example, cells expressing EphA2 (e.g., cells collected by biopsy of a tumor or metastatic lesion; cells from an established cancer cell line; or recombinant cells) can be cultured in vitro in a culture medium, and the contacting step can be affected by adding the ADC to the culture medium. The method will result in the killing of cells expressing EphA2, particularly including cancer cells expressing EphA2. Alternatively, the ADC can be administered to a subject by any suitable route of administration (e.g., intravenously, subcutaneously, or in direct contact with tumor tissue) to act in vivo.

[1133] The in vivo effects of the disclosed ADC therapeutic compositions can be evaluated in suitable animal models. For example, xenogeneic cancer models can be used, in which cancer explants or passaged xenograft tissues are introduced into immunocompromised animals, such as nude mice or SCID mice (Klein et al. (1997) Nature Med. 3: 402-8). Efficacy can be predicted using assays that measure inhibition of tumor formation, tumor regression, or metastasis, etc.

[1134] In vivo assays can assess the contribution of mechanisms such as apoptosis to tumor death. In some embodiments, xenografts from tumor-bearing mice treated with the therapeutic composition can be examined for the presence of apoptotic foci and compared to untreated control xenograft-bearing mice. The extent to which apoptotic foci are found in tumors from treated mice provides an indication of the therapeutic efficacy of the composition.

[1135] Further provided herein are methods for treating disorders (e.g., cancer). The compositions described herein, such as the ADCs disclosed herein, can be administered to a non-human mammal or a human subject for therapeutic purposes. The treatment methods comprise administering to a subject having or suspected of having cancer a therapeutically effective amount of a composition, such as an ADC, comprising a Bcl-xL inhibitor, wherein the inhibitor is linked to a targeting antibody that binds to an antigen that (1) is expressed on cancer cells, (2) is readily bound, and / or (3) is localized or predominantly expressed on the surface of cancer cells compared to non-cancerous cells.

[1136] Exemplary embodiments are methods for treating a subject suffering from or suspected of having cancer, comprising administering to a subject a therapeutically effective amount of a composition disclosed herein, such as an ADC, composition, or pharmaceutical composition (such as any exemplary ADC, composition, or pharmaceutical composition disclosed herein). In some embodiments, cancer expresses a target antigen. In some embodiments, cancer is a tumor or a blood cancer. In some embodiments, cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, 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 (including acute lymphocytic 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, gastric cancer, colon cancer, or head and neck cancer. In some embodiments, cancer is lymphoma or gastric cancer. In some embodiments, the cancer is breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, stomach cancer, bladder cancer, or colon cancer. In some embodiments, the cancer is breast cancer or non-small cell lung cancer.

[1137] Another exemplary embodiment is a method of delivering a Bcl-xL inhibitor to a cell expressing EphA2, the method comprising conjugating the Bcl-xL inhibitor to an antibody that immunospecifically binds to an EphA2 epitope and exposing the cell to the ADC. Exemplary EphA2-expressing cancer cells indicated by the ADC of the present disclosure include breast cancer cells, non-small cell lung cancer cells, pancreatic cancer cells, esophageal cancer cells, head and neck cancer cells, gastric cancer cells, bladder cancer cells, or colon cancer cells.

[1138] In certain aspects, the present disclosure also provides a method for reducing or inhibiting the growth of a tumor (e.g., a tumor expressing EphA2), 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 the growth of a patient's tumor, reduce the number or size of metastatic lesions, reduce tumor burden, reduce primary tumor burden, reduce invasiveness, prolong survival time, and / or maintain or improve quality of life. 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-EphA2 antibody) when administered alone, and / or the tumor is resistant or refractory to treatment with a Bcl-xL inhibitor drug moiety when administered alone.

[1139] An exemplary embodiment is a method of reducing or inhibiting tumor growth in a subject, the method comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any exemplary ADC, composition, or pharmaceutical composition disclosed herein). In some embodiments, the tumor expresses a target antigen. 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, pancreatic cancer, gastric cancer, colon cancer, or spleen cancer. In some embodiments, the tumor is gastric cancer. In some embodiments, the tumor is breast cancer, non-small cell lung cancer, pancreatic cancer, esophageal cancer, head and neck cancer, gastric cancer, bladder cancer, or colon cancer. In some embodiments, the tumor is breast cancer or non-small cell lung cancer. In some embodiments, administration of the ADC, composition, or pharmaceutical composition reduces or inhibits the growth of a tumor by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about, at least 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.

[1140] Another exemplary embodiment is a method for delaying or slowing tumor growth in a subject, the method comprising administering to the subject a therapeutically effective amount of an ADC, composition, or pharmaceutical composition (e.g., any exemplary ADC, composition, or pharmaceutical composition disclosed herein). In some embodiments, the tumor expresses a target antigen. 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 spleen cancer. In some embodiments, the tumor is gastric cancer. In some embodiments, the administration of an ADC, composition, or pharmaceutical composition delays or slows the growth of the tumor by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about, at least 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 growth without treatment.

[1141] In certain aspects, the present disclosure further provides methods of reducing or slowing the expansion of a cancer cell population (eg, a cancer cell population expressing EphA2), the method comprising administering a therapeutically effective amount of an ADC or a composition comprising an ADC.

[1142] Exemplary embodiments are methods for reducing or slowing down the expansion of cancer cell groups in a subject, the method comprising administering a therapeutically effective amount of ADC, composition, or pharmaceutical composition (such as any exemplary ADC, composition, or pharmaceutical composition disclosed herein) to the subject. In some embodiments, the cancer cell group expresses a target antigen. In some embodiments, the cancer cell group is from a tumor or blood cancer. In some embodiments, the cancer cell group is from breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, 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 (including acute lymphocytic leukemia), follicular lymphoma, T cell or B cell derived lymphoid malignancies, melanoma, myeloid leukemia, myeloma, oral cancer, ovarian cancer, non-small cell lung cancer, prostate cancer, small cell lung cancer, spleen cancer, or head and neck cancer. In some embodiments, the cancer cell group is from lymphoma or gastric cancer. In some embodiments, administration of an 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 without treatment. In some embodiments, administration of an ADC, composition, or pharmaceutical composition slows the expansion of a 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 expansion in the absence of treatment.

[1143] Also provided herein are methods for determining whether a subject having 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 having or suspected of having cancer will respond to treatment with an ADC, composition, or pharmaceutical composition (e.g., any exemplary ADC, composition, or pharmaceutical composition disclosed herein) by providing a biological sample from a 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 a 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 amplification of cancer cell populations in culture, etc.).

[1144] Further provided herein are therapeutic uses of the disclosed ADCs and compositions. An exemplary embodiment is an ADC, composition, or pharmaceutical composition (e.g., any exemplary ADC, composition, or pharmaceutical composition disclosed herein) for treating a subject suffering from or suspected of having cancer (e.g., a cancer expressing EphA2). Another exemplary embodiment is the use of an ADC, composition, or pharmaceutical composition (e.g., any exemplary ADC, composition, or pharmaceutical composition disclosed herein) for treating a subject suffering from or suspected of having cancer (e.g., a cancer expressing EphA2). Another exemplary embodiment is the use of an ADC, composition, or pharmaceutical composition (e.g., any exemplary ADC, composition, or pharmaceutical composition disclosed herein) in a method for manufacturing a medicament for treating a subject suffering from or suspected of having cancer (e.g., a cancer expressing EphA2). Methods for identifying subjects suffering from cancer expressing a target antigen (e.g., EphA2) are known in the art and can be used to identify patients suitable for treatment with a disclosed ADC compound or composition.

[1145] In addition, for veterinary purposes or as animal models of human disease, the ADCs of the present disclosure can be administered to non-human mammals that express an antigen to which the ADC is capable of binding. In the latter case, such animal models can be used to evaluate the therapeutic efficacy of the disclosed ADCs (e.g., testing doses and administration schedules).

[1146] The therapeutic composition for implementing the aforementioned method can be formulated into a pharmaceutical composition 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, such as a carrier suitable for selected modes of administration (such as intravenous administration). The pharmaceutical composition can also include one or more other inactive agents and / or therapeutic agents (such as standard nursing agents, etc.) suitable for treating or preventing cancer, such as cancer. The pharmaceutical composition can also include one or more carriers, excipients, and / or stabilizer components, etc. The method for preparing such pharmaceutical compositions and suitable formulations is known in the art (see, for example, "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, PA).

[1147] Suitable carriers include any material that retains the anti-tumor function of the therapeutic composition when combined with the therapeutic composition and that generally does not react with the patient's immune system. Pharmaceutically acceptable carriers include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and absorption delaying agents that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, and mesylate, and combinations thereof. In many cases, isotonic agents, such as sugars, polyols (such as mannitol, sorbitol), or sodium chloride, are included in the composition. Pharmaceutically acceptable carriers may further include a small amount of auxiliary substances, such as wetting agents or emulsifiers, preservatives, or buffers, which enhance the shelf life or effectiveness of the ADC.

[1148] The pharmaceutical composition of the present disclosure can be applied by various methods known in the art. The route of administration and / or mode can vary according to the desired result. In some embodiments, the therapeutic preparation is dissolved and applied by any route that can deliver 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, in situ, etc. In some embodiments, application is intravenous, subcutaneous, intraperitoneal or intramuscular. Pharmaceutically acceptable carriers should be suitable for routes of administration, such as intravenous or subcutaneous administration (e.g., by injection or infusion). Depending on the route of administration, the active compound, i.e., ADC and / or any other therapeutic agent, can be encapsulated in a material to protect the compound from the effects of acid and other natural conditions that may inactivate the compound. Application can be systemic or local.

[1149] The therapeutic compositions disclosed herein can be sterile and stable under manufacturing and storage conditions, and can be in various forms. These include, for example, liquid, semisolid and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersants 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 ADC can b...

Claims

1. An antibody-drug conjugate of formula (1): Ab-(LD) p (1) wherein Ab is an anti-EphA2 antibody or an antigen-binding fragment thereof; L is a linker that covalently attaches Ab to D; p is an integer from 1 to 16; and D is a Bcl-xL inhibitor compound of Formula (I) or Formula (II) covalently attached to the linker L: or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein: R1 and R2 independently represent a group selected from the following: hydrogen; a straight or branched C1-C6 alkyl group, which is optionally substituted by a hydroxyl or C1-C6 alkoxy group; a C3-C6 cycloalkyl group; a trifluoromethyl group; a straight or branched C1-C6 alkylene-heterocycloalkyl group, wherein the heterocycloalkyl group is optionally substituted by a straight or branched C1-C6 alkyl group; or R1 and R2 together with the carbon atom carrying them form a C3-C6 cycloalkylene group, R3 represents a group selected from the following: hydrogen; C3-C6 cycloalkyl; straight chain or branched C1-C6 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ;-X1-OR c ;-X1-COOR c ; -X1-PO(OH)2; -X1-SO2(OH); -X1-N3 and: R a and R b independently of one another represent a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl; a straight-chain or branched C1-C6 alkyl optionally substituted by one or two hydroxyl groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; Ca-C6 alkylene-NR d R e ; C1-C6 alkylene-N + R d R e R f ; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a C1-C6 alkoxy group; Group: or R a and R b Together with the nitrogen atom that carries them, they form ring B1; or R a , R b and R c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R c , R d , R e , R f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R d and R e Together with the nitrogen atom carrying them, they form ring B2, or R d , R e and R f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, Het1 represents a group selected from the following: Het2 represents a group selected from the following: A1 is -NH-, -N(C1-C3 alkyl), O, S or Se, A2 is N, CH or C(R5), G is selected from the group consisting of: -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , C1-C6 alkyl optionally substituted by hydroxyl groups, halogen, -NO2 and -CN, wherein: -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2)- 1-4 -phenyl; or R G1 and R G2 and their respective atoms to which they are attached are combined to form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of: Where R G4 is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl optionally substituted by 1 to 3 halogen atoms, R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group, R5 represents a group selected from the following: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; C2-C6 alkenyl; C2-C6 alkynyl; halogen or -CN, R6 represents a group selected from the following: hydrogen; -C2-C6 alkenyl; -X2-O-R7; -X2-NSO2-R7; -C=C(R9)-Y1-O-R7; C3-C6 cycloalkyl; C3-C6 heterocycloalkyl optionally substituted by a hydroxy group; C3-C6 cycloalkylene-Y2-R7; C3-C6 heterocycloalkylene-Y2-R7 group, a heteroarylene-R7 group optionally substituted by a straight-chain or branched C1-C6 alkyl group, R7 represents a group selected from the following: a straight or branched C1-C6 alkyl group; (C3-C6) cycloalkylene-R8; or: Wherein Cy represents a C3-C8 cycloalkyl group, R8 represents a group selected from the following: hydrogen; straight chain or branched C1-C6 alkyl, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R' c ;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b 、-X'2-NR' a R' b 、-NR' c -X'2-N3 and: R9 represents a group selected from a linear or branched C1-C6 alkyl group, a trifluoromethyl group, a hydroxyl group, a halogen group, a C1-C6 alkoxy group, R 10 represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl, R 11 represents a group selected from hydrogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8, C3-C8 heterocycloalkylene-CH2-R8, R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group, R 14 and R 15 independently of one another represent hydrogen or a methyl group, or R 14 and R 15 Together with the carbon atoms that carry them, they form a cyclohexyl group, R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group, X1 and X2 independently represent a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy, X'2 represents a straight chain or branched C1-C6 alkylene group, R' a and R' b represents independently of one another a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl group; a straight-chain or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ; C1-C6 alkylene-N + R′ d R′ e R′ f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a hydroxyl group or a C1-C6 alkoxy group; Group: or R' a and R' b Together with the nitrogen atom carrying them, they form ring B3, or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R' c , R' d , R' e , R' f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R' d and R' e Together with the nitrogen atom carrying them, they form ring B4, or R′ d , R′ e and R′ f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, Y1 represents a straight or branched C1-C4 alkylene group, Y2 represents bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-, -C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-, m = 0, 1 or 2, p = 1, 2, 3 or 4, B1, B2, B3 and B4 independently represent a C3-C8 heterocycloalkyl group, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH2, oxo or piperidinyl, wherein R3 and, if present, one of the R8 groups are covalently attached to the linker and wherein the valence of the atom is not exceeded by one or more substituents bonded thereto; or or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein: n = 0, 1 or 2, ------ indicates a single bond or a double bond, A4 and A5 independently represent a carbon or nitrogen atom, Z1 represents a bond, -N(R)- or -O-, wherein R represents hydrogen or a linear or branched C1-C6 alkyl group, R1 represents a group selected from the following: hydrogen; a straight or branched C1-C6 alkyl group optionally substituted by a hydroxyl or C1-C6 alkoxy group; a C3-C6 cycloalkyl group; a trifluoromethyl group; a straight or branched C1-C6 alkylene-heterocycloalkyl group, wherein the heterocycloalkyl group is optionally substituted by a straight or branched C1-C6 alkyl group; R2 represents hydrogen or methyl; R3 represents a group selected from the following: hydrogen; straight chain or branched C1-C4 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ;-X1-OR c ;-X1-COOR c ; -X1-PO(OH)2; -X1-SO2(OH); -X1-N3 and: R a and R b independently of one another represent a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl; a straight-chain or branched C1-C6 alkyl optionally substituted by one or two hydroxyl groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR d R e ; C1-C6 alkylene-N + R d R e R f ; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a C1-C6 alkoxy group; Group: or R a and R b Together with the nitrogen atom that carries them, they form ring B1; or R a , R b and R c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R c , R d , R e , R f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R d and R e Together with the nitrogen atom carrying them, they form ring B2, or R d , R e and R f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, Het1 represents a group selected from the following: Het2 represents a group selected from the following: A1 is -NH-, -N(C1-C3 alkyl), O, S or Se, A2 is N, CH or C(R5), G is selected from the group consisting of: -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , C1-C6 alkyl optionally substituted by hydroxyl groups, halogen, -NO2 and -CN, wherein: -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2)- 1-4 -phenyl; or R G1 and R G2 and their respective atoms to which they are attached are combined to form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of: Where R G4 is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl optionally substituted by 1 to 3 halogen atoms, R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group, R5 represents a group selected from the following: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; C2-C6 alkenyl; C2-C6 alkynyl; halogen or -CN, R6 represents a group selected from the following: hydrogen; -C2-C6 alkenyl; -X2-O-R7; -X2-NSO2-R7; -C=C(R9)-Y1-O-R7; C3-C6 cycloalkyl; C3-C6 heterocycloalkyl optionally substituted by a hydroxy group; C3-C6 cycloalkylene-Y2-R7; C3-C6 heterocycloalkylene-Y2-R7 group, a heteroarylene-R7 group optionally substituted by a straight-chain or branched C1-C6 alkyl group, R7 represents a group selected from the following: a straight or branched C1-C6 alkyl group; (C3-C6) cycloalkylene-R8; or: Wherein Cy represents a C3-C8 cycloalkyl group, R8 represents a group selected from the following: hydrogen; straight chain or branched C1-C6 alkyl, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R'c;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b 、-X'2-NR' a R' b 、-NR' c -X'2-N3 and: R9 represents a group selected from a linear or branched C1-C6 alkyl group, a trifluoromethyl group, a hydroxyl group, a halogen group, a C1-C6 alkoxy group, 10 represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl, R 11 represents a group selected from hydrogen, halogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8, C3-C8 heterocycloalkylene-CH2-R8, R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group, R 14 and R 15 independently of one another represent hydrogen or methyl, or R 14 and R 15 Together with the carbon atoms that carry them, they form a cyclohexyl group, R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group, X1 represents a straight or branched C1-C4 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxyl, halogen, C1-C6 alkoxy, X2 represents a straight or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxyl, halogen, C1-C6 alkoxy, X'2 represents a straight chain or branched C1-C6 alkylene group, R' a and R' b represents independently of one another a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl group; a straight-chain or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ; C1-C6 alkylene-N + R′ d R′ e R′ f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a hydroxyl group or a C1-C6 alkoxy group; Group: or R' a and R' b Together with the nitrogen atom carrying them, they form ring B3, or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R' c , R' d , R' e , R' f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R' d and R' e Together with the nitrogen atom carrying them, they form ring B4, or R′ d , R′ e and R′ f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, Y1 represents a straight or branched C1-C4 alkylene group, Y2 represents bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-, -C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-, m = 0, 1 or 2, p = 1, 2, 3 or 4, B1, B2, B3 and B4 independently represent a C3-C8 heterocycloalkyl group, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH2, oxo or piperidinyl, wherein R3 and, if present, one of the R8 groups are covalently attached to the linker, and wherein the valency of the atom is not exceeded by one or more substituents bonded thereto; and The anti-EphA2 antibody or antigen-binding fragment comprises: (a) three heavy chain complementarity determining regions (HCDRs) comprising the amino acid sequences of SEQ ID NO: 2 (HCDR1), SEQ ID NO: 3 (HCDR2), and SEQ ID NO: 4 (HCDR3); and three light chain complementarity determining regions (LCDRs) comprising the amino acid sequences of SEQ ID NO: 12 (LCDR1), SEQ ID NO: 13 (LCDR2), and SEQ ID NO: 14, 26, 29, 32, or 35 (LCDR3); (b) three heavy chain complementarity determining regions (HCDRs) comprising the amino acid sequences of SEQ ID NO: 5 (HCDR1), SEQ ID NO: 6 (HCDR2), and SEQ ID NO: 4 (HCDR3); and three light chain complementarity determining regions (LCDRs) comprising the amino acid sequences of SEQ ID NO: 15 (LCDR1), SEQ ID NO: 16 (LCDR2), and SEQ ID NO: 17, 24, 27, 30, 33, or 36 (LCDR3); (c) three heavy chain complementarity determining regions (HCDRs) comprising the amino acid sequences of SEQ ID NO: 7 (HCDR1), SEQ ID NO: 8 (HCDR2) and SEQ ID NO: 9 (HCDR3); and three light chain complementarity determining regions (LCDRs) comprising the amino acid sequences of SEQ ID NO: 18 (LCDR1), SEQ ID NO: 13 (LCDR2) and SEQ ID NO: 17, 24, 27, 30, 33 or 36 (LCDR3); or (d) three heavy chain complementarity determining regions (HCDRs) comprising the amino acid sequences of SEQ ID NO: 10 (HCDR1), SEQ ID NO: 6 (HCDR2) and SEQ ID NO: 4 (HCDR3); and three light chain complementarity determining regions (LCDRs) comprising the amino acid sequences of SEQ ID NO: 15 (LCDR1), SEQ ID NO: 16 (LCDR2) and SEQ ID NO: 17, 27, 30, 33 or 36 (LCDR3).

2. The antibody-drug conjugate of claim 1, wherein p is an integer from 1 to 6 or from 2 to 4, or p is 2 or 4; or p is determined by liquid chromatography-mass spectrometry (LC-MS).

3. The antibody-drug conjugate of claim 1 or 2, wherein L comprises: Attachment groups; at least one bridging spacer group; and At least one cleavable group, optionally at least one cleavable group comprising a pyrophosphate group and / or a self-immolative group.

4. The antibody-drug conjugate of claim 3, wherein -(LD) has formula (A): in: R 1 is an attachment group; L1 is a bridging spacer group; E is a cleavable group.

5. The antibody-drug conjugate of claim 3 or 4, wherein the cleavable group comprises a pyrophosphate group or the cleavable group comprises 6. The antibody-drug conjugate of claim 3 or 4, wherein the bridging spacer group comprises: (i) a polyoxyethylene (PEG) group; (ii) a PEG group selected from the group consisting of PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, and PEG15; (iii) -CO-CH2-CH2-PEG12- group; (iv) a butyryl, pentanoyl, hexanoyl, heptanoyl or octanoyl group; or (v) Hexanoyl group.

7. The antibody-drug conjugate of claim 6, wherein (i) the attachment group is formed by at least one reactive group selected from a maleimide group, a thiol group, a cyclooctyne group and an azide group; optionally wherein: a) the maleimide group has the structure: b) The azido group has the following structure: -N=N + =N - ; c) the cyclooctyne group has the structure: and wherein -* is a bond to said antibody; or d) the cyclooctyne group has the structure: and wherein -* is a bond to said antibody; or (ii) the attachment group has a formula comprising: and Where -* is a bond to the antibody.

8. The antibody-drug conjugate of claim 7, wherein the antibody is joined to the linker (L) via an attachment group selected from: wherein -* is a bond to the antibody, and wherein is a bond to the bridging spacer group.

9. The antibody-drug conjugate of claim 8, wherein the bridging spacer group is -CO-CH2-CH2-PEG12-.

10. The antibody-drug conjugate of claim 8 or 9, wherein the bridging spacer group is joined to a cleavable group; optionally the cleavable group is -pyrophosphate-CH2-CH2-NH2-.

11. The antibody-drug conjugate of any one of claims 8 to 10, wherein the cleavable group is conjugated to a Bcl-xL inhibitor (D).

12. The antibody-drug conjugate of any one of claims 1 to 3, wherein the linker comprises: Attachment group, at least one bridging spacer group, Peptide groups, and At least one cleavable group.

13. The antibody-drug conjugate of claim 12, wherein -(LD) has formula (B): in: R 1 is an attachment group; L1 is a bridging spacer; Lp is a peptide group comprising 1 to 6 amino acid residues, or Lp comprises the group E is a cleavable group L2 is a bridging spacer; m is 0 or 1; and D is a Bcl-xL inhibitor.

14. The antibody-drug conjugate of claim 12 or 13, wherein (i) the attachment group is formed by at least one reactive group comprising a maleimide group, a thiol group, a cyclooctyne group and / or an azide group, optionally wherein: a) the maleimide group has the structure: b) The azido group has the structure: -N=N + =N - ;or c) the cyclooctyne group has the structure: and wherein -* is a bond to said antibody; or (ii) the attachment group has a formula comprising: and Where -* is a bond to the antibody.

15. The antibody-drug conjugate of any one of claims 12 to 14, wherein: (i) at least one bridging spacer comprises a PEG group, optionally the PEG group is selected from PEG1, PEG2, PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14 and PEG15; or (ii) at least one bridging spacer is selected from *-C(O)-CH2-CH2-PEG1-**, *-C(O)-CH2-PEG3-**, *-C(O)-CH2-CH2-PEG12**, *-NH-CH2-CH2-PEG1-**, a polyhydroxyalkyl group, *-C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)-**, and *-C(O)-CH2-CH2-PEG12-NH-C(O)CH2-CH2-**, wherein ** represents the direct or indirect attachment point of the at least one bridging spacer to the attachment group, and * represents the direct or indirect attachment point of the at least one bridging spacer to the peptide group.

16. The antibody-drug conjugate of any one of claims 12 to 15, wherein L1 is selected from *-C(O)-CH2-CH2-PEG1-**, *-C(O)-CH2-PEG3-**, *-C(O)-CH2-CH2-PEG12**, *-NH-CH2-CH2-PEG1-** and a polyhydroxyalkyl group, wherein ** represents the direct or indirect point of attachment of L1 to R1, and * represents the direct or indirect point of attachment of L1 to Lp.

17. The antibody-drug conjugate of any one of claims 12 to 16, wherein m is 1 and L2 is -C(O)-N(CH3)-CH2-CH2-N(CH3)-C(O)-.

18. The antibody-drug conjugate of any one of claims 12 to 17, wherein (i) the peptide group comprises 1 to 6, 1 to 4, 1 to 3 or 1 to 2 amino acid residues, optionally the amino acid residues are selected from 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); (ii) the peptide group comprises Val-Cit, Val-Ala, Val-Lys, Sulfo-Ala-Val-Cit, Sulfo-Ala-Val-Ala, Gly-Gly-Gly and / or Gly-Gly-Phe-Gly (SEQ ID NO: 78); or (iii) the peptide group is selected from:

19. The antibody-drug conjugate of any one of claims 12 to 18, wherein (i) the cleavable group comprises a pyrophosphate group and / or a self-immolative group; (ii) the cleavable group comprises a self-immolative group; or (iii) the cleavable group comprises a self-immolative group, the self-immolative group comprising p-aminobenzyl-carbamate, p-aminobenzyl-ammonium, p-amino-(sulfo)benzyl-ammonium, p-amino-(sulfo)benzyl-carbamate, p-amino-(alkoxy-PEG-alkyl)benzyl-carbamate, p-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-carbamate or p-amino-(polyhydroxycarboxytetrahydropyranyl)alkyl-benzyl-ammonium.

20. The antibody-drug conjugate of any one of claims 13 to 19, wherein m is 0 or 1 or m is 1, and the bridging spacer comprises 21. The antibody-drug conjugate of any one of claims 13 to 20, wherein -(LD) is formed by a compound selected from the group consisting of:

22. The antibody drug conjugate of any one of claims 13 to 21, wherein -(LD) comprises a formula selected from: Where -* is a bond to the antibody.

23. The antibody-drug conjugate of claim 1 or 2, wherein -(LD) has formula (C): in: R 1 is an attachment group; L1 is a bridging spacer; L p is a peptide group containing 1 to 6 amino acids; D is a Bcl-xL inhibitor; G1-L2-A is a self-immolative spacer; L2 is a bond, methylene, neopentylene or C2-C3 alkenylene; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a is independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; L3 is a spacer moiety; and R 2 It is the hydrophilic part.

24. The antibody-drug conjugate of claim 23, or a pharmaceutically acceptable salt thereof, wherein -(LD) has formula (D): in: R 1 is an attachment group; L1 is a bridging spacer; Lp is a peptide group containing 1 to 6 amino acids; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a is independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; L3 is a spacer moiety; and R 2 It is the hydrophilic part.

25. The antibody-drug conjugate of claim 23 or 24, wherein: (1) L1 includes: *-CH(OH)CH(OH)CH(OH)CH(OH)-**, wherein each n is an integer from 1 to 12, wherein the * of L1 indicates a direct or indirect point of attachment to Lp, and the ** of L1 indicates a point of attachment to R 1 direct or indirect attachment point; (2) L1 is n is an integer from 1 to 12 or n is 1 or n is 12, wherein the * of L1 indicates a direct or indirect attachment point to Lp, and the ** of L1 indicates a direct or indirect attachment point to R 1 direct or indirect attachment point; (3) L1 is and n is an integer from 1 to 12, wherein the * of L1 indicates a direct or indirect attachment point to Lp, and the ** of L1 indicates a direct or indirect attachment point to R 1 direct or indirect attachment point; (4) L1 contains Where * of L1 indicates the direct or indirect attachment point to Lp, and ** of L1 indicates the attachment point to R 1 direct or indirect point of attachment; or (5) L1 is a bridging spacer comprising: *-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 3 C(=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 -**, where * of L1 indicates the direct or indirect attachment point to Lp, and ** of L1 indicates the attachment point to R 1 direct or indirect attachment point; X1 is 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; and 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; And each R 3 Independently selected from H and C1-C6 alkyl.

26. The antibody-drug conjugate of any one of claims 23 to 25, wherein R 2 The hydrophilic part comprises polyethylene glycol, polyalkylene glycol, polyol, polysarcosine, sugar, oligosaccharide, polypeptide, 1 to 3 substituted C2-C6 alkyl, or substituted by 1 to 2 independently selected from -OC(=O)NHS(O)2NHCH2CH2OCH3, -NHC(=O)C 1-4 Alkylene -P(O)(OCH2CH3)2 and -COOH group substituted C2-C6 alkyl.

27. The antibody-drug conjugate of any one of claims 23 to 26, wherein R 2 yes Where n is an integer from 1 to 6, 28. The antibody-drug conjugate of claim 23 or 24, wherein the hydrophilic portion comprises: (i) Polysarcosine having the following moieties: wherein n is an integer from 3 to 25; and R is H, -CH3 or -CH2CH2C(=O)OH; or (ii) polyethylene glycol of the formula: Where R is H, -CH3, CH2CH2NHC(=O)OR a 、-CH2CH2NHC(=O)R a or -CH2CH2C(=O)OR a , R' is OH, -OCH3, -CH2CH2NHC(=O)OR a 、-CH2CH2NHC(=O)R a or-OCH2CH2C(=O)OR a , where R a is H or optionally OH or C 1-4 Alkoxy substituted C 1-4 An alkyl group, and m and n are each independently an integer from 2 to 25.

29. The antibody-drug conjugate of any one of claims 23 to 27, wherein the hydrophilic portion comprises 30. The antibody-drug conjugate of any one of claims 23 to 29, wherein: (i) L3 has the structure The spacer portion of in: 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 is independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl; and X is a bond, triazolyl or -CH2-triazolyl-, Where X is connected to R 2 ;or (ii) L3 has the structure The spacer portion of in: 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 is independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl; and X is -CH2-triazolyl-C 1-4 Alkylene-OC(O)NHS(O)2NH-, -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 -, -CH2-triazolyl-C 1-4 Alkylene-OC(O)NHS(O)2NH-(CH2CH2O) n -、-C 4-6 Cycloalkylene-OC(O)NHS(O)2NH-(CH2CH2O) n -, wherein each n is independently 1, 2 or 3, Where X is connected to R 2 .

31. The antibody-drug conjugate of any one of claims 3 to 30, wherein the attachment group is formed by a reaction involving at least one reactive group.

32. The antibody-drug conjugate of any one of claims 3 to 31, wherein the attachment group is formed by reacting: a first reactive group attached to the linker, and a second reactive group attached to the antibody or as an amino acid residue of the antibody, wherein optionally, (i) at least one of the reactive groups comprises: Mercaptans, Maleimide, Haloacetamide, Azide, Alkynes, Cyclooctene, Triarylphosphine, Oxanorbornadiene, Cyclooctyne, Diaryltetrazine, Monoaryl tetrazines, Norbornene, aldehyde, Hydroxylamine, Hydrazine, NH2-NH-C(=O)-, ketone, Vinyl sulfone, Aziridine, Amino acid residues, -ONH2、-NH2、 -N3、 -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、 -C(O)NHNH2, in: Each R 3 Independently selected from H and C1-C6 alkyl; Each R 4 is 2-pyridyl or 4-pyridyl; Each R 5 Independently selected from H, C1-C6 alkyl, F, Cl and -OH; Each R 6 Independently selected from H, C1-C6 alkyl, F, Cl, -NH2, -OCH3, -OCH2CH3, -N(CH3)2, -CN, -NO2 and -OH; Each R 7 Independently selected from H, C 1-6 alkyl, fluorine, benzyloxy substituted by -C(=O)OH, benzyl substituted by -C(=O)OH, C(=O) 1-4 Alkoxy and C substituted by -C(=O)OH 1-4 Alkyl; and / or (ii) the first reactive group and the second reactive group comprise: Thiol and maleimide, Thiols and haloacetamides, Thiol and vinyl sulfone, Thiols and aziridines, Azides and alkynes, Azide and cyclooctyne, Azide and cyclooctene, Azides and triarylphosphines, Azides and oxa-norbornadiene, Diaryltetrazine and cyclooctene, Monoaryl tetrazines and norbornene, Aldehydes and hydroxylamines, Aldehydes and hydrazines, Aldehydes and NH2-NH-C(=O)-, Ketones and hydroxylamines, Ketones and hydrazines, Ketone and NH2-NH-C(=O)-, Hydroxylamine and Amine and or CoA or a CoA analog and a serine residue.

33. The antibody-drug conjugate of any one of claims 3 to 32, wherein the attachment group comprises a group selected from: Amide; as well as Disulfide, in: R 32 It is H, C 1-4 Alkyl, phenyl, pyrimidine or pyridine; R 35 It is H, C 1-6 Alkyl, phenyl or C substituted with 1 to 3 -OH groups 1-4 alkyl; Each R 7 Independently selected from H, C 1-6 alkyl, fluorine, benzyloxy substituted with -C(=O)OH, benzyl substituted with -C(=O)OH, C(=O) 1-4 Alkoxy and C substituted by -C(=O)OH 1-4 alkyl; R 37 independently selected from H, phenyl and pyridine; q is 0, 1, 2, or 3; R 8 is H or methyl; and R 9 It is H, -CH3 or phenyl.

34. The antibody-drug conjugate of any one of claims 23 to 33, wherein the peptide group comprises 1 to 4 or 1 to 3 or 1 or 2 amino acid residues, optionally selected from 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).

35. The antibody-drug conjugate of any one of claims 23 to 33, wherein the peptide group comprises Val-Cit, Phe-Lys, Val-Ala, Val-Lys, Leu-Cit, Sulfo-Ala-Val-Cit, Sulfo-Ala-Val-Ala, Gly-Gly-Gly and / or Gly-Gly-Phe-Gly (SEQ ID NO: 78).

36. The antibody-drug conjugate of any one of claims 23 to 35, wherein Lp is selected from:

37. The antibody-drug conjugate of any one of claims 23 to 36, wherein: -(LD) comprises or is formed from a compound of the formula: in: R is H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: R is H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: R is H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: Each R is independently selected from H, -CH3 and -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: Each R is independently selected from H, -CH3 and -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: X a is -CH2-, -OCH2-, -NHCH2- or -NRCH2- and each R is independently H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: R is H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: Xb is -CH2-, -OCH2-, -NHCH2-, or -NRCH2- and each R is independently H, -CH3, or -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; in: A is a bond, -OC(=v)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor; or in: Each R is independently H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor, or in: Each R is independently H, -CH3 or -CH2CH2C(=O)OH; A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a is independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; n is an integer from 2 to 24; and D is a Bcl-xL inhibitor, or in: A is a bond, -OC(=O)-*, -OC(=O)N(CH3)CH2CH2N(CH3)C(=O)-*or-OC(=O)N(CH3)C(R a )2C(R a )2N(CH3)C(=O)-*, Each R a are independently selected from H, C1-C6 alkyl and C3-C8 cycloalkyl, and the * of A represents the point of attachment to D; and D is a Bcl-xL inhibitor.

38. The antibody-drug conjugate of any one of claims 23 to 37, wherein A is a bond and / or R is -CH3 or -CH2CH2COOH.

39. The antibody-drug conjugate of any one of claims 23 to 37, wherein A is -OC(=O)-* and / or R is -CH3 or -CH2CH2COOH.

40. The antibody-drug conjugate of any one of claims 23 to 39, wherein -(LD) is formed by a compound selected from:

41. The antibody-drug conjugate of any one of claims 1 to 40, wherein D comprises a compound of formula (I): or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein: R1 and R2 independently represent a group selected from the following: hydrogen; a straight or branched C1-C6 alkyl group, which is optionally substituted by a hydroxyl or C1-C6 alkoxy group; a C3-C6 cycloalkyl group; a trifluoromethyl group; a straight or branched C1-C6 alkylene-heterocycloalkyl group, wherein the heterocycloalkyl group is optionally substituted by a straight or branched C1-C6 alkyl group; or R1 and R2 together with the carbon atom carrying them form a C3-C6 cycloalkylene group, R3 represents a group selected from the following: hydrogen; C3-C6 cycloalkyl; straight chain or branched C1-C6 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ;-X1-OR c ;-X1-COOR c ; -X1-PO(OH)2; -X1-SO2(OH); -X1-N3 and: R a and R b independently of one another represent a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl; a straight-chain or branched C1-C6 alkyl optionally substituted by one or two hydroxyl groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR d R e ; C1-C6 alkylene-N + R d R e R f ; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a C1-C6 alkoxy group; Group: or R a and R b Together with the nitrogen atom that carries them, they form ring B1; or R a , R b and R c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R c , R d , R e , R f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R d and R e Together with the nitrogen atom carrying them, they form ring B2, or R d , R e and R f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, Het1 represents a group selected from the following: Het2 represents a group selected from the following: A1 is -NH-, -N(C1-C3 alkyl), O, S or Se, A2 is N, CH or C(R5), G is selected from the group consisting of: -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , C1-C6 alkyl optionally substituted by hydroxyl groups, halogen, -NO2 and -CN, wherein: -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2)- 1-4 -phenyl; or R G1 and R G2 and their respective atoms to which they are attached are combined to form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of: Where R G4 is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl optionally substituted by 1 to 3 halogen atoms, R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group, R5 represents a group selected from the following: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; C2-C6 alkenyl; C2-C6 alkynyl; halogen or -CN, R6 represents a group selected from the following: hydrogen; -C2-C6 alkenyl; -X2-O-R7; -X2-NSO2-R7; -C=C(R9)-Y1-O-R7; C3-C6 cycloalkyl; C3-C6 heterocycloalkyl optionally substituted by a hydroxy group; C3-C6 cycloalkylene-Y2-R7; C3-C6 heterocycloalkylene-Y2-R7 group, a heteroarylene-R7 group optionally substituted by a straight-chain or branched C1-C6 alkyl group, R7 represents a group selected from the following: a straight chain or branched C1-C6 alkyl group; (C3-C6)cycloalkylene-R8; or: Wherein Cy represents a C3-C8 cycloalkyl group, R8 represents a group selected from the following: hydrogen; straight chain or branched C1-C6 alkyl, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R' c ;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b 、-X'2-NR' a R' b 、-NR' c -X'2-N3 and: R9 represents a group selected from a linear or branched C1-C6 alkyl group, a trifluoromethyl group, a hydroxyl group, a halogen group, a C1-C6 alkoxy group, 10 represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl, R 11 represents a group selected from hydrogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8, C3-C8 heterocycloalkylene-CH2-R8, R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group, R 14 and R 15 independently of one another represent hydrogen or a methyl group, or R 14 and R 15 Together with the carbon atoms that carry them, they form a cyclohexyl group, R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group, X1 and X2 independently represent a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy, X'2 represents a straight chain or branched C1-C6 alkylene group, R' a and R' b represents independently of one another a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl group; a straight-chain or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ; C1-C6 alkylene-N + R′ d R′ e R′ f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a hydroxyl group or a C1-C6 alkoxy group; Group: or R' a and R' b Together with the nitrogen atom carrying them, they form ring B3, or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R' c , R' d , R' e , R' f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R' d and R' e Together with the nitrogen atom carrying them, they form ring B4, or R′ d , R′ e and R′ f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, Y1 represents a straight or branched C1-C4 alkylene group, Y2 represents bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-, -C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-, m = 0, 1 or 2, p = 1, 2, 3 or 4, B1, B2, B3 and B4 independently represent a C3-C8 heterocycloalkyl group, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH2, oxo or piperidinyl, wherein R3 and, if present, one of the R8 groups are covalently attached to the linker, and wherein the valency of the atom is not exceeded by one or more substituents bonded thereto.

42. The antibody-drug conjugate of claim 41, wherein R1 is a linear or branched C 1-6 Alkyl and R2 is H.

43. The antibody-drug conjugate of any one of claims 1 to 40, wherein D comprises a compound of formula (II): or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein: n = 0, 1 or 2, ------ indicates a single bond or a double bond, A4 and A5 independently represent a carbon or nitrogen atom, Z1 represents a bond, -N(R)- or -O-, wherein R represents hydrogen or a linear or branched C1-C6 alkyl group, R1 represents a group selected from the following: hydrogen; a straight or branched C1-C6 alkyl group optionally substituted by a hydroxyl or C1-C6 alkoxy group; a C3-C6 cycloalkyl group; a trifluoromethyl group; a straight or branched C1-C6 alkylene-heterocycloalkyl group, wherein the heterocycloalkyl group is optionally substituted by a straight or branched C1-C6 alkyl group; R2 represents hydrogen or methyl; R3 represents a group selected from the following: hydrogen; straight chain or branched C1-C4 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ; -X1-O-R c ; -X1-COOR c ; -X1-PO(OH)2; -X1-SO2(OH); -X1-N3 and: R a and R b independently of one another represent a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl; a straight-chain or branched C1-C6 alkyl optionally substituted by one or two hydroxyl groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR d R e ; C1-C6 alkylene-N + R d R e R f ; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a C1-C6 alkoxy group; Group: or R a and R b Together with the nitrogen atom that carries them, they form ring B1; or R a , R b and R c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R c , R d , R e , R f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R d and R e Together with the nitrogen atom carrying them, they form ring B2, or R d , R e and R f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, Het1 represents a group selected from the following: Het2 represents a group selected from the following: A1 is -NH-, -N(C1-C3 alkyl), O, S or Se, A2 is N, CH or C(R5), G is selected from the group consisting of: -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , C1-C6 alkyl optionally substituted by hydroxyl groups, halogen, -NO2 and -CN, wherein: -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2)- 1-4 -phenyl; or R G1 and R G2 and their respective atoms to which they are attached are combined to form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of: Where R G4 is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl optionally substituted by 1 to 3 halogen atoms, R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group, R5 represents a group selected from the following: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; C2-C6 alkenyl; C2-C6 alkynyl; halogen or -CN, R6 represents a group selected from the following: hydrogen; -C2-C6 alkenyl; -X2-O-R7; -X2-NSO2-R7; -C=C(R9)-Y1-O-R7; C3-C6 cycloalkyl; C3-C6 heterocycloalkyl optionally substituted by a hydroxy group; C3-C6 cycloalkylene-Y2-R7; C3-C6 heterocycloalkylene-Y2-R7 group, a heteroarylene-R7 group optionally substituted by a straight-chain or branched C1-C6 alkyl group, R7 represents a group selected from the following: a straight chain or branched C1-C6 alkyl group; (C3-C6)cycloalkylene-R8; or: Wherein Cy represents a C3-C8 cycloalkyl group, R8 represents a group selected from the following: hydrogen; straight chain or branched C1-C6 alkyl, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R'c;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b 、-X'2-NR' a R' b 、-NR' c -X'2-N3 and: R9 represents a group selected from a linear or branched C1-C6 alkyl group, a trifluoromethyl group, a hydroxyl group, a halogen group, a C1-C6 alkoxy group, 10 represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl, R 11 represents a group selected from hydrogen, halogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8, C3-C8 heterocycloalkylene-CH2-R8, R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group, R 14 and R 15 independently of one another represent hydrogen or methyl, or R 14 and R 15 Together with the carbon atoms that carry them, they form a cyclohexyl group, R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group, X1 represents a straight or branched C1-C4 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxyl, halogen, C1-C6 alkoxy, X2 represents a straight or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxyl, halogen, C1-C6 alkoxy, X'2 represents a straight chain or branched C1-C6 alkylene group, R' a and R' b represents independently of one another a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl group; a straight-chain or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ; C1-C6 alkylene-N + R′ d R′ e R′ f ; C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a hydroxyl group or a C1-C6 alkoxy group; Group: or R' a and R' b Together with the nitrogen atom carrying them, they form ring B3, or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R' c , R' d , R' e , R' f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R' d and R' e Together with the nitrogen atom carrying them, they form ring B4, or R′ d , R′ e and R′ f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, Y1 represents a straight or branched C1-C4 alkylene group, Y2 represents bond, -O-, -O-CH2-, -O-CO-, -O-SO2-, -CH2-, -CH2-O, -CH2-CO-, -CH2-SO2-, -C2H5-, -CO-, -CO-O-, -CO-CH2-, -CO-NH-CH2-, -SO2-, -SO2-CH2-, -NH-CO-, -NH-SO2-, m = 0, 1 or 2, p = 1, 2, 3 or 4, B1, B2, B3 and B4 independently represent a C3-C8 heterocycloalkyl group, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH2, oxo or piperidinyl, wherein R3 and, if present, one of the R8 groups are covalently attached to the linker, and wherein the valency of the atom is not exceeded by one or more substituents bonded thereto.

44. The antibody-drug conjugate of claim 43, wherein A1 and A5 both represent nitrogen atoms, and R1 is a linear or branched C 1-6 Alkyl; R2 is H; n is 1; and ------ represents a single bond.

45. The antibody-drug conjugate of any one of claims 1 to 44, wherein G is selected from the group consisting of: -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , halogen, -NO2 and -CN, among which: -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen, C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2) 1-4 -phenyl; -R G3 is selected from the group consisting of C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, phenyl and -(CH2)- 1-4 -phenyl; or R G1 and R G2 and their respective atoms to which they are attached are combined to form a C3-C8 heterocycloalkyl; or in the alternative, G is selected from the group consisting of: Where R G4 Selected from C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl and C3-C6 cycloalkyl which are optionally substituted by 1 to 3 halogen atoms.

46. ​​The antibody-drug conjugate of any one of claims 1 to 40, wherein D comprises a compound of formula (IA) or (IIA): or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein: Z1 represents a bond or -O-, R3 represents a group selected from the following: hydrogen; C3-C6 cycloalkyl; straight chain or branched C1-C6 alkyl; -X1-NR a R b ; -X1-N + R a R b R c ; and -X1-O-R c , R a and R b independently of one another represent a group selected from the group consisting of: hydrogen; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl groups; and a C1-C6 alkylene group -SO2O - , R c represents hydrogen or a linear or branched C1-C6 alkyl group, Het2 represents a group selected from the following: A1 is -NH-, -N(C1-C3 alkyl), O, S or Se, A2 is N, CH or C(R5), G is selected from the group consisting of: -C(O)OH、-C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , C1-C6 alkyl optionally substituted by hydroxyl groups, halogen, -NO2 and -CN, wherein: -R G1 and R G2 is independently selected at each occurrence from the group consisting of hydrogen and C1-C6 alkyl optionally substituted with 1 to 3 halogen atoms; -R G3 is C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; or R G1 and R G2 Together with the atoms to which they are attached, they are combined to form a C3-C8 heterocycloalkyl; R4 represents a hydrogen, fluorine, chlorine or bromine atom, a methyl, hydroxyl or methoxy group, R5 represents a group selected from the following: C1-C6 alkyl optionally substituted by 1 to 3 halogen atoms; halogen or -CN, R6 represents a group selected from the following: -X2-O-R7; and a heteroarylene-R7 group optionally substituted by a straight-chain or branched C1-C6 alkyl group, R7 represents a group selected from the following: a straight or branched C1-C6 alkyl group; (C3-C6) cycloalkylene-R8; or: Wherein Cy represents a C3-C8 cycloalkyl group, R8 represents a group selected from the following: hydrogen; straight chain or branched C1-C6 alkyl, -NR' a R' b ;-NR' a -CO-OR' c ;-NR' a -CO-R' c ;-N + R' a R' b R' c ;-O-R' c ;-NH-X'2-N + R' a R' b R' c ;-O-X'2-NR' a R' b ;-X'2-NR' a R' b :-NR' c -X'2-N3 and: R 10 represents a group selected from hydrogen, fluorine, chlorine, bromine, -CF3 and methyl, R 11 represents a group selected from hydrogen, C1-C3 alkylene-R8, -O-C1-C3 alkylene-R8, -CO-NR h R i and -CH=CH-C1-C4 alkylene-NR h R i , -CH=CH-CHO, C3-C8 cycloalkylene-CH2-R8, C3-C8 heterocycloalkylene-CH2-R8, R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group, R 14 and R 15 independently of one another represent hydrogen or a methyl group, or R 14 and R 15 Together with the carbon atoms that carry them, they form a cyclohexyl group, R h and R i independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group, X1 and X2 independently represent a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy, X'2 represents a straight chain or branched C1-C6 alkylene group, R' a and R' b represents independently of one another a group selected from the following: hydrogen; heterocycloalkyl; -SO2-phenyl, wherein the phenyl may be substituted by a straight-chain or branched C1-C6 alkyl group; a straight-chain or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl or C1-C6 alkoxy groups; C1-C6 alkylene-SO2OH; C1-C6 alkylene-SO2O - ; C1-C6 alkylene-COOH; C1-C6 alkylene-PO(OH)2; C1-C6 alkylene-NR' d R' e ; C1-C6 alkylene-N + R′ d R′ e R′ f .C1-C6 alkylene-O-C1-C6 alkylene-OH; C1-C6 alkylene-phenyl, wherein the phenyl group may be substituted by a hydroxyl group or a C1-C6 alkoxy group; Group: or R' a and R' b Together with the nitrogen atom carrying them, they form ring B3, or R' a , R' b and R' c C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, R' c , R' d , R' e , R' f independently represent hydrogen or a linear or branched C1-C6 alkyl group, or R' d and R' e Together with the nitrogen atom carrying them, they form ring B4, or R′ d , R′ e and R′ f C3-C8 heterocycloalkyl groups forming a bridge with the nitrogen atom carrying them, m = 0, 1 or 2, p = 1, 2, 3 or 4, B3 and B4 independently represent a C3-C8 heterocycloalkyl group, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spiro ring system, (ii) in addition to the nitrogen atom, may also contain one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl, -NH2, oxo or piperidinyl.

47. The antibody-drug conjugate of claim 46, wherein G is selected from the group consisting of: -C(O)OH, -C(O)OR G3 、-C(O)NR G1 R G2 、-C(O)R G2 、-NR G1 C(O)R G2 、-NR G1 C(O)NR G1 R G2 、-OC(O)NR G1 R G2 、-NR G1 C(O)OR G3 、-C(=NOR G1 )NR G1 R G2 、-NR G1 C(=NCN)NR G1 R G2 、-NR G1 S(O)2NR G1 R G2 、-S(O)2R G3 、-S(O)2NR G1 R G2 、-NR G1 S(O)2R G2 、-NR G1 C(=NR G2 )NR G1 R G2 、-C(=S)NR G1 R G2 、-C(=NR G1 )NR G1 R G2 , halogen, -NO2 and -CN.

48. The antibody-drug conjugate of any one of claims 1 to 47, wherein R7 represents a group selected from the following: a linear or branched C1-C a Alkyl group; (C3-C a ) cycloalkylene-R8; or: wherein Cy represents a C3-C8 cycloalkyl group.

49. The antibody-drug conjugate of any one of claims 1 to 47, wherein R7 represents a group selected from the following:

50. The antibody-drug conjugate of any one of claims 1 to 40, wherein D comprises a compound of formula (IB), (IC), (IIB) or (IIC): or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof, wherein: For formula (IB) or (IC), R3 represents a group selected from the following: hydrogen; linear or branched C1-C6 alkyl; -X1-NR a R b ;-X1-N + R a R b R c ; and-X1-OR c ; For formula (IIB) or (IIC), Z1 represents a bond and R3 represents hydrogen; or Z1 represents -O- and R3 represents -X1-NR a R b , R a and R b independently of one another represent a group selected from the group consisting of: hydrogen; a linear or branched C1-C6 alkyl group optionally substituted by one or two hydroxyl groups; and a C1-C6 alkylene group -SO2O - , R c represents hydrogen or a straight or branched C1-C6 alkyl group R6 represents -X2-O-R7 or a heteroarylene-R7 group optionally substituted by a linear or branched C1-C6 alkyl group, R7 represents a group selected from the following: R8 represents a group selected from the following: -NR' a R' b ;-O-X'2-NR' a R' b ; and -X'2-NR' a R' b , R 10 represents fluorine, R 12 and R 13 independently of one another represent a hydrogen atom or a methyl group, R 14 and R 15 independently of one another represent hydrogen or a methyl group, X1 and X2 independently represent a linear or branched C1-C6 alkylene group, which is optionally substituted by one or two groups selected from trifluoromethyl, hydroxy, halogen, C1-C6 alkoxy, X'2 represents a straight chain or branched C1-C6 alkylene group, R' a and R' b independently represent a group selected from the following: hydrogen; a straight or branched C1-C6 alkyl group optionally substituted with one or two hydroxyl or C1-C6 alkoxy groups; a C1-C6 alkylene group -NR' d R' e ; or R' a and R' b Together with the nitrogen atom carrying them, they form ring B3, R' d , R' e independently of one another represent hydrogen or a linear or branched C1-C6 alkyl group, B3 represents a C3-C8 heterocycloalkyl group, which may: (i) be a monocyclic group or a bicyclic group, wherein the bicyclic group includes a fused, bridged or spirocyclic system, (ii) in addition to the nitrogen atom, may also contain one or two heteroatoms independently selected from oxygen and nitrogen, (iii) be substituted by one or two groups selected from the following: fluorine, bromine, chlorine, straight or branched C1-C6 alkyl, hydroxyl and oxo.

51. The antibody-drug conjugate of any one of claims 1 to 50, wherein R7 represents the following group:

52. The antibody-drug conjugate of any one of claims 1 to 50, wherein R7 represents a group selected from:

53. The antibody-drug conjugate of any one of claims 41 to 52, wherein R8 represents a group selected from: wherein --* represents a bond to the linker.

54. The antibody-drug conjugate of any one of claims 41 to 53, wherein B3 represents a C3-C8 heterocycloalkyl group selected from a pyrrolidinyl group, a piperidinyl group, a piperazinyl group, a morpholinyl group, an azepanyl group and a 4,4-difluoropiperidin-1-yl group.

55. The antibody-drug conjugate of any one of claims 1 to 40, wherein D represents any of the following attached to L: or any enantiomer, diastereomer and / or pharmaceutically acceptable salt thereof.

56. The antibody-drug conjugate of any one of claims 1 to 40, wherein D comprises a group represented by a formula selected from those in Table A2.

57. The antibody-drug conjugate of any one of claims 1 to 40, wherein -(LD) is formed from a compound in Table B or an enantiomer, diastereomer and / or pharmaceutically acceptable salt of any of the foregoing.

58. The antibody-drug conjugate of any one of claims 1 to 57, wherein: The anti-EphA2 antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 11, 19, 20, 21, 22, 23, 25, 28, 31, 34, 71, 72 or 73.

59. The antibody-drug conjugate of any one of claims 1 to 57, wherein the anti-EphA2 antibody or antigen-binding fragment comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 37 and a light chain comprising the amino acid sequence of SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 75, SEQ ID NO: 76 or SEQ ID NO:

77.

60. The antibody-drug conjugate of any one of claims 1 to 57, wherein the anti-EphA2 antibody or antigen-binding fragment comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 39 and a light chain comprising the amino acid sequence of SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 75, SEQ ID NO: 76 or SEQ ID NO:

77.

61. The antibody-drug conjugate of any one of claims 1 to 57, wherein the anti-EphA2 antibody or antigen-binding fragment comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 74 and a light chain comprising the amino acid sequence of SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 75, SEQ ID NO: 76 or SEQ ID NO:

77.

62. The antibody-drug conjugate of claim 60, wherein the anti-EphA2 antibody or antigen-binding fragment comprises: A heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO:

41.

63. The antibody-drug conjugate of claim 61, wherein the anti-EphA2 antibody or antigen-binding fragment comprises: A heavy chain comprising the amino acid sequence of SEQ ID NO:74, and a light chain comprising the amino acid sequence of SEQ ID NO:

41.

64. A composition comprising multiple copies of the antibody-drug conjugate of any one of claims 1 to 63, wherein the average p of the antibody-drug conjugates in the composition is about 2 to about 16, such as about 2 to about 8, such as about 2 to about 4.

65. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 1 to 63 or the composition of claim 64, and a pharmaceutically acceptable carrier.

66. A method of treating a subject having or suspected of having cancer, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 63, the composition of claim 64, or the pharmaceutical composition of claim 65.

67. The method of claim 66, wherein the cancer expresses EphA2.

68. The method of claim 66 or 67, wherein the cancer is a tumor or a blood cancer, optionally the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, 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, including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer or head and neck cancer.

69. The method of claim 68, wherein the cancer is breast cancer or non-small cell lung cancer.

70. A method of reducing or inhibiting tumor growth in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 63, the composition of claim 64, or the pharmaceutical composition of claim 65.

71. The method of claim 70, wherein the tumor expresses EphA2.

72. The method of claim 70 or 71, wherein 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, pancreatic cancer, stomach cancer, colon cancer, or spleen cancer.

73. The method of claim 72, wherein the tumor is breast cancer or non-small cell lung cancer.

74. A method of reducing or inhibiting a blood cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 63, the composition of claim 64, or the pharmaceutical composition of claim 65.

75. The method of claim 74, wherein the hematological cancer expresses EphA2.

76. The method of claim 74 or 75, wherein the blood cancer is chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), Hodgkin's lymphoma, or non-Hodgkin's lymphoma or myelodysplastic syndrome (MDS).

77. The method of any one of claims 70 to 76, wherein administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces or inhibits tumor growth or blood cancer 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%.

78. A method of reducing or slowing the expansion of a cancer cell population in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate as described in any one of claims 1 to 63, a composition as described in claim 64, or a pharmaceutical composition as described in claim 63.

79. The method of claim 78, wherein the cancer cell population expresses EphA2.

80. The method of claim 78 or 79, wherein the cancer cell population is from a tumor or blood cancer, optionally wherein the cancer cell population is from breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, 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 including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer, or head and neck cancer.

81. The method of claim 80, wherein the cancer cell population is from breast cancer or non-small cell lung cancer.

82. The method of any one of claims 78 to 81, wherein administration of the antibody-drug conjugate, composition, or pharmaceutical composition reduces the cancer cell population or 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%.

83. The method of any one of claims 66 to 82, wherein the antibody-drug conjugate is administered as a monotherapy.

84. The method of any one of claims 66 to 82, wherein the antibody-drug conjugate is administered adjunctively with another therapeutic agent or radiation therapy.

85. The method of claim 84, wherein the antibody-drug conjugate is administered in an amount effective to sensitize the tumor cells to one or more additional therapeutic agents and / or radiation therapy.

86. The method of any one of claims 66 to 82, further comprising administering at least one additional therapeutic agent to a subject in need thereof.

87. The method of claim 86, wherein the one additional therapeutic agent is a Bcl-2 inhibitor, a taxane, a vinca alkaloid, a MEK inhibitor, an ERK inhibitor, a topoisomerase inhibitor, a nucleoside analog, or a RAF inhibitor.

88. The method of claim 86, wherein the one additional therapeutic agent is selected from venetoclax, Compound A2, vincristine, topotecan, docetaxel, paclitaxel, LTT463, trametinib, gemcitabine, and LXH254.

89. A method of inhibiting Bcl-xL activity in a cell expressing Bcl-xL, the method comprising contacting the cell with an antibody-drug conjugate of any one of claims 1 to 63 capable of binding to the cell under conditions wherein the antibody drug conjugate binds to the cell.

90. A method for determining whether a subject having or suspected of having cancer will respond to treatment with an antibody-drug conjugate as described in any one of claims 1 to 63, a composition as described in claim 64, or a pharmaceutical composition as described in claim 65, the method comprising providing a biological sample from the subject; contacting the sample with the antibody-drug conjugate; and detecting binding of the antibody-drug conjugate to cancer cells in the sample.

91. The method of claim 90, wherein the cancer cells in the sample express EphA2.

92. The method of claim 90 or claim 91, wherein the cancer expresses EphA2.

93. The method of any one of claims 90 to 92, wherein the cancer is a tumor or a blood cancer, optionally the cancer is breast cancer, multiple myeloma, plasma cell myeloma, leukemia, lymphoma, sarcoma, 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, including acute 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, prostate cancer, small cell lung cancer, spleen cancer, pancreatic cancer, stomach cancer, colon cancer or head and neck cancer.

94. The method of claim 93, wherein the cancer is breast cancer or non-small cell lung cancer.

95. The method of any one of claims 90 to 94, wherein the sample is a tissue biopsy sample, a blood sample, or a bone marrow sample.

96. A method of preparing an antibody-drug conjugate as claimed in any one of claims 1 to 63, the method comprising reacting an anti-EphA2 antibody or antigen-binding fragment with a cleavable linker conjugated to a Bcl-xL inhibitor under conditions that permit conjugation.

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