Splicing modulator antibody-drug conjugates and methods of use thereof

Abstract

To provide linker-drug compounds and antibody-drug conjugates that bind to human oncology targets.SOLUTION: The linker-drug compounds and antibody-drug conjugates of the present disclosure comprise a splicing modulator drug moiety. The present disclosure further relates to methods and compositions for use in the treatment of neoplastic disorders by administering the antibody-drug conjugates provided herein. In certain embodiments, the splicing modulator comprises a pladienolide or a pladienolide derivative.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This disclosure claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 679,672, filed June 1, 2018; U.S. Provisional Patent Application No. 62 / 679,631, filed June 1, 2018; and U.S. Provisional Patent Application No. 62 / 779,324, filed December 13, 2018. All of the foregoing applications are incorporated herein by reference in their entirety.

[0002] The present disclosure relates to antibody-drug conjugates (ADCs) comprising a splicing modulator and an antibody or antigen-binding fragment thereof that binds to a human oncology antigen target. The disclosure further relates to methods and compositions useful in the treatment or diagnosis of cancers that express the target antigen and / or are amenable to treatment by disruption of RNA splicing, as well as methods of making such compositions. [Background technology]

[0003] The majority of protein-coding genes in the human genome consist of multiple exons (coding regions) separated by introns (non-coding regions). Gene expression results in a single precursor messenger RNA (pre-mRNA). Intron sequences are then removed from the pre-mRNA in a process called splicing, resulting in the mature messenger RNA (mRNA). Alternative splicing results in the inclusion of various combinations of exons, resulting in mRNAs that encode distinct protein isoforms.

[0004] RNA splicing is catalyzed by the spliceosome, a dynamic multiprotein-RNA complex composed of five small nuclear RNAs (snRNAs U1, U2, U4, U5, and U6) and associated proteins. The spliceosome assembles on pre-mRNAs and orchestrates a dynamic cascade of multiple RNA and protein interactions that catalyze intron excision and exon ligation (Matera and Wang (2014) Nat Rev Mol Cell Biol. 15(2):108-21). Increasing evidence links human diseases to aberrant regulation of RNA splicing, affecting many genes (Scotti and Swanson (2016) Nat Rev Genet. 17(1):19-32).

[0005] The spliceosome is an important target in cancer biology. Several studies have now demonstrated significant changes in the splicing profile of cancer cells as well as in the splicing factors themselves (Agrawal et al. (2018) Curr Opin Genet Dev. 48:67-74). Alternative splicing can lead to exon inclusion / exclusion, intron retention, or differential utilization of cryptic splice sites (Seiler et al. (2018) Cell Rep. 23(1):282-296). Collectively, these events describe functional changes that may contribute to tumor development or therapeutic resistance (Siegfried and Karni (2018) Curr Opin Genet Dev. 48:16-21).

[0006] Certain natural products can bind to the SF3b spliceosome complex. These small molecules modulate splicing by enhancing intron retention and / or exon skipping (Teng et al. (2017) Nat Commun. 8:15522). A significant proportion of the resulting transcripts contain premature termination codons that trigger nonsense-mediated mRNA decay (NMD). Furthermore, impaired canonical splicing leads to a significant reduction in canonical transcripts, which can negatively impact cell function and viability. Therefore, splicing modulators are a promising class of drugs for the treatment of cancer (Puthenveetil et al. (2016) Bioconjugate Chem. 27:1880-8).

[0007] The proto-oncogene human epidermal growth factor receptor 2 (HER2) encodes a transmembrane tyrosine kinase receptor belonging to the human epidermal growth factor receptor (EGFR) family (King et al. (1985) Science 229:974-6). Overexpression of HER2 allows constitutive activation of growth factor signaling pathways, such as the PI3K-AKT-mTOR pathway, and thus serves as an oncogenic driver in several cancer types, including approximately 20% of invasive breast cancers (Slamon et al. (1989) Science 244:707-12; Gajria and Chandarlapaty (2011) Expert Rev Anticancer Ther. 11:263-75). Given that HER2 amplification mediates a transformed phenotype, and because HER2 expression is largely restricted to malignant cells, HER2 is a promising antigen for targeting certain cancers and / or delivering novel cancer therapies (Parakh et al. (2017) Cancer Treat Rev. 59:1-21). Additional antigens for targeted cancer therapy delivery include, but are not limited to, CD138 (also known as syndecan-1) and ephrin type-A receptor 2 (EPHA2).

[0008] CD138 is a cell surface heparan sulfate proteoglycan essential for maintaining cell morphology and interacting with the surrounding microenvironment (Akl et al. (2015) Oncotarget 6(30):28693-715; Szatmari et al. (2015) Dis Markers 2015:796052). Overall, loss of CD138 expression in cancer cells reduces cell adhesion to the extracellular matrix and enhances cell motility and invasion (Teng et al. (2012) Matrix Biol. 31:3-16). Increased stromal CD138 expression also alters fibronectin production and extracellular matrix organization (Yang et al. (2011) Am J Pathol. 178:325-35). Additionally, increased CD138 expression on stromal fibroblasts is associated with angiogenesis and cancer progression (Maeda et al. (2006) Oncogene 25:1408-12). CD138 expression increases during B cell development, and its presence is a hallmark of plasma cells (Ribatti (2017) Immunol Lett. 188:64-7). CD138 expression is maintained in multiple myeloma, a plasma cell malignancy. Therefore, CD138 is an attractive antigen for targeted therapy of several cancers and other hematological malignancies (Sherbenou et al. (2015) Blood Rev. 29(2):81-91; Wijdenes et al. (1996) Br J Haematol. 94(2):318-23).

[0009] EPHA2 is a transmembrane glycoprotein that is highly overexpressed in several malignant cancer-derived cell lines and advanced cancers (Wykosky and Debinski (2008) Mol Cancer Ref. 6(12):1795-1806). For example, EPHA2 is strongly overexpressed in approximately 61% of GBM patient tumors (Wykosky et al. (2008) Clin Cancer Res. 14:199-208), 76% of ovarian cancers (Thaker et al. (2004) Clin Cancer Res. 10:5145-50), and 85% of prostate adenocarcinomas (Zeng et al. (2003) Am J Pathol. 163:2271-6). The EPHA2 protein is highly overexpressed in a proportion of patient tumors and in a proportion of cells within tumors and is a cell membrane-localized receptor that can internalize upon ligand binding (Walker-Daniels et al. (2002) Mol Cancer Res. 1:79-87). Furthermore, EPHA2 expression is associated with poor prognosis, increased metastasis, and reduced survival. Therefore, because of its expression pattern, localization, and functional importance in cancer patient outcome, EPHA2 is another attractive antigen for the delivery of novel targeted anticancer therapies. Summary of the Invention [Means for solving the problem]

[0010] In various embodiments, the present disclosure provides, in part, novel compounds having antineoplastic cellular biological activity. The compounds may slow, inhibit, and / or reverse tumor growth in mammals and may be useful in treating human cancer patients. In various embodiments, the present disclosure provides novel antibody-drug conjugates that employ the novel compounds or other functional splice inhibitor molecules.

[0011] More specifically, in various embodiments, the present disclosure relates to antibody-drug conjugate (ADC) compounds capable of binding to and killing neoplastic cells. In various embodiments, the ADC compounds disclosed herein include a linker connecting a splicing modulator to a full-length antibody or antigen-binding fragment. In various embodiments, the ADC compounds also have the ability to internalize into target cells after binding.

[0012] In various embodiments, the ADC compound has formula (I): Ab-(LD) p (I) where Ab is an antibody or antigen-binding fragment thereof that targets a neoplastic cell or another oncology-related target; D is a splicing modulator; L is a linker covalently linking Ab to D; and p is an integer from 1 to 15. It can be represented by:

[0013] In various embodiments, the ADC compound has formula (I): Ab-(LD) p (I) where Ab is an antibody or antigen-binding fragment thereof that targets a neoplastic cell; D is a group represented by formula (II): [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1 is selected from absent, hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 3is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; and R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR 17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1 to C6 alkyl group, and -NR 15 R 16 Selected from; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 Selected from; R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; and Z is [ka] Selected from; R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 at least one of is hydrogen; L is a linker covalently linking Ab to D; and p is an integer from 1 to 15. It can be represented by:

[0014] In some embodiments, the antibody or antigen-binding fragment has the ability to internalize into a target cell. In some embodiments, the linker is covalently attached to a splicing modulator of formula (II) ("LD"), wherein LD has formula (II-A): [ka] (Wherein Z' is [ka] Selected from; All other variables are as defined for formula (II). or a pharmaceutically acceptable salt thereof.

[0015] In various other embodiments, the ADC compound has formula (I): Ab-(LD) p (I) where Ab is an antibody or antigen-binding fragment thereof that targets a neoplastic cell; D is a compound of formula (IV): [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, an -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; and R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR 17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1 to C6 alkyl group, and -NR 15 R 16 Selected from; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 selected from; and R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 15 , R 16 , and R17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 at least one of is hydrogen; L is a linker covalently linking Ab to D; and p is an integer from 1 to 15. It can be represented by:

[0016] In some embodiments, the antibody or antigen-binding fragment has the ability to internalize into a target cell. In some embodiments, the linker is covalently attached to the splicing modulator ("LD"), and the LD has the formula (IV-A): [ka] or a pharmaceutically acceptable salt thereof.

[0017] In various other embodiments, the ADC compound has formula (I): Ab-(LD) p (I) where Ab is an antibody or antigen-binding fragment thereof that targets a neoplastic cell; D is a compound of formula (VI): [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1 and R 9are each independently selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR 17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1-C6 alkyl group, -NR 15 R 16 and a linker; R 10 is selected from hydrogen, a C1-C6 alkyl group, a -C(=O)-(C1-C6 alkyl) group, and -CD3; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 Selected from; R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; and a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; R 1, R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 at least one of is hydrogen; R 1 and R 9 cannot both exist; L is a linker covalently linking Ab to D; and p is an integer from 1 to 15. It can be represented by:

[0018] In some embodiments, the antibody or antigen-binding fragment has the ability to internalize into a target cell. In some embodiments, the linker is covalently attached to the splicing modulator ("LD"), and the LD has the formula (VI-A): [ka] or a pharmaceutically acceptable salt thereof.

[0019] In various other embodiments, the ADC compound has formula (I): Ab-(LD) p (I) where Ab is an antibody or antigen-binding fragment thereof that targets a neoplastic cell; D is a group represented by formula (VIII): [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1 is selected from absent, hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; R 4 is selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —O—C(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; and R 10 is selected from a 3- to 10-membered carbocyclic ring and a 3- to 10-membered heterocyclic ring, each of which is selected from 0 to 3 R a Each R is replaced by a are independently a halogen, a C1-C6 alkyl group, an -O-(C1-C6) alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylhydroxy group, or -S(=O) w -(4- to 7-membered heterocycle), a 4- to 7-membered carbocycle, and a 4- to 7-membered heterocycle; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 selected from; and R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; R 1 , R3 , R 4 , R 10 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; Each R a are independently a halogen, a hydroxyl group, or -NR 15 R 16 , C1-C6 alkyl group, -(C=O)-(C1-C6 alkyl) group, -(C=O)-(C1-C6 alkyl)-(C3-C 10 heterocyclyl group), -S(=O) w -(C3-C8 heterocyclyl) group, and C1-C6 alkylcarboxylic acid group (wherein each of these is selected from the group consisting of halogen, hydroxyl, -NR 15 R 16 and C1-C3 alkyl; w is 0, 1, or 2; L is a linker covalently linking Ab to D; and p is an integer from 1 to 15. It can be represented by:

[0020] In some embodiments, the antibody or antigen-binding fragment has the ability to internalize into a target cell. In some embodiments, the linker is covalently attached to the splicing modulator ("LD"), and the LD has the formula (VIII-A): [ka] or a pharmaceutically acceptable salt thereof.

[0021] In some embodiments, the splicing modulator comprises a modulator of the SF3b complex. In some embodiments, the splicing modulator comprises a pladienolide or a pladienolide derivative. In some embodiments, the splicing modulator comprises pladienolide D or a pladienolide D derivative. In some embodiments, the pladienolide D or derivative comprises D2, D1, D4, D8, D10, D11 (E7107), D20, D21, D22, D12, or D25. In some embodiments, the pladienolide D or derivative comprises D2. In some embodiments, the pladienolide D or derivative comprises D1. In some embodiments, the pladienolide D or derivative comprises D4. In some embodiments, the pladienolide D or derivative comprises D12.

[0022] In some embodiments, the pladienolide D or derivative is a zwitterionic pladienolide D or derivative. In some embodiments, the zwitterionic pladienolide D or derivative comprises D22 or D25.

[0023] In some other embodiments, the splicing modulator comprises pladienolide B or a pladienolide B derivative. In some embodiments, the pladienolide B or derivative comprises D9, D18, D19, or D13. In some embodiments, the splicing modulator comprises an arylpladienolide. In some embodiments, the arylpladienolide comprises D15, D14, D16, D17, D26, or D33. In some embodiments, the arylpladienolide comprises D15. In some embodiments, the arylpladienolide is a zwitterionic arylpladienolide. In some embodiments, the zwitterionic arylpladienolide comprises D33.

[0024] In some embodiments, the splicing modulator is D1: [ka] Includes.

[0025] In some embodiments, the splicing modulator is D2: [ka] Includes.

[0026] In some embodiments, the splicing modulator is D3: [ka] Includes.

[0027] In some embodiments, the splicing modulator is D4: [ka] Includes.

[0028] In some embodiments, the splicing modulator is D4': [ka] Includes.

[0029] In some embodiments, the splicing modulator is D5: [ka] Includes.

[0030] In some embodiments, the splicing modulator is D6: [ka] Includes.

[0031] In some embodiments, the splicing modulator is D7: [ka] Includes.

[0032] In some embodiments, the splicing modulator is D8: [ka] Includes.

[0033] In some embodiments, the splicing modulator is D9: [ka] Includes.

[0034] In some embodiments, the splicing modulator is D10: [ka] Includes.

[0035] In some embodiments, the splicing modulator is D11: [ka] Includes.

[0036] In some embodiments, the splicing modulator is D12: [ka] Includes.

[0037] In some embodiments, the splicing modulator is D13: [ka] Includes.

[0038] In some embodiments, the splicing modulator is D14: [ka] Includes.

[0039] In some embodiments, the splicing modulator is D15: [ka] Includes.

[0040] In some embodiments, the splicing modulator is D16: [ka] Includes.

[0041] In some embodiments, the splicing modulator is D17: [ka] Includes.

[0042] In some embodiments, the splicing modulator is D18: [ka] Includes.

[0043] In some embodiments, the splicing modulator is D19: [ka] Includes.

[0044] In some embodiments, the splicing modulator is D20: [ka] Includes.

[0045] In some embodiments, the splicing modulator is D21: [ka] Includes.

[0046] In some embodiments, the splicing modulator is D22: [ka] Includes.

[0047] In some embodiments, the splicing modulator is D23: [ka] Includes.

[0048] In some embodiments, the splicing modulator is D24: [ka] Includes.

[0049] In some embodiments, the splicing modulator is D25: [ka] Includes.

[0050] In some embodiments, the splicing modulator is D26: [ka] Includes.

[0051] In some embodiments, the splicing modulator is D27: [ka] Includes.

[0052] In some embodiments, the splicing modulator is D28: [ka] Includes.

[0053] In some embodiments, the splicing modulator is D29: [ka] Includes.

[0054] In some embodiments, the splicing modulator is D30: [ka] Includes.

[0055] In some embodiments, the splicing modulator is D31: [ka] Includes.

[0056] In some embodiments, the splicing modulator is D32: [ka] Includes.

[0057] In some embodiments, the splicing modulator is D33: [ka] Includes.

[0058] In some embodiments, the splicing modulator is D34: [ka] Includes.

[0059] In some embodiments, the splicing modulator is D35: [ka] Includes.

[0060] In some embodiments, the splicing modulator comprises one of the drug moieties listed in Table 7. In some embodiments, the splicing modulator comprises D1, D2, D3, D4, D4', D5, D6, D7, D8, D9, D10, D11, D12, D13, D14, D15, D16, D17, D18, D19, D20, D21, D22, D23, D24, D25, D26, D27, D28, D29, D30, D31, D32, D33, D34, and / or D35.

[0061] In some embodiments, splicing modulators and their use as therapeutic agents alone or as part of an ADC are disclosed. In some embodiments, the splicing modulators comprise D4, D4', D12, D15, D8, D9, D10, D13, D18, D19, D20, D21, D22, D25, or D33.

[0062] In some embodiments, the splicing modulator comprises D4 and the linker comprises MC-Val-Cit-pABC. In some embodiments, the splicing modulator comprises D4 and the linker comprises MC-β-glucuronide. In some embodiments, the splicing modulator comprises D12 and the linker comprises MC-Val-Cit-pABC. In some embodiments, the splicing modulator comprises D12 and the linker comprises MC-β-glucuronide. In some embodiments, the splicing modulator comprises D15 and the linker comprises MC-Val-Ala-pAB.

[0063] In various embodiments, the linkers used in the ADCs disclosed herein are stable outside of a cell, such that the ADC remains intact when present in extracellular conditions, but is capable of cleavage upon internalization into a cell, e.g., a tumor or cancer cell. In some embodiments, the splicing modulator is cleaved from the antibody or antigen-binding fragment when the ADC enters a cell expressing the antigen targeted by the antibody or antigen-binding fragment of the ADC. In some embodiments, the linker is a cleavable linker.

[0064] In some embodiments, the linker comprises a cleavable peptide moiety. In some embodiments, the cleavable peptide moiety is cleavable by an enzyme. In some embodiments, the cleavable peptide moiety or linker comprises an amino acid unit. In some embodiments, the amino acid unit comprises valine-citrulline ("Val-Cit" or "VC"). In some other embodiments, the amino acid unit comprises valine-alanine ("Val-Ala" or "VA"). In some other embodiments, the amino acid unit comprises glutamic acid-valine-citrulline ("Glu-Val-Cit" or "EVC"). In some other embodiments, the amino acid unit comprises alanine-alanine-asparagine ("Ala-Ala-Asn" or "AAN").

[0065] In some embodiments, the linker comprises a cleavable glucuronide moiety. In some embodiments, the cleavable glucuronide moiety is cleavable by an enzyme. In some embodiments, the cleavable glucuronide moiety is cleavable by a glucuronidase. In some embodiments, the cleavable glucuronide moiety is cleavable by a β-glucuronidase.

[0066] In some embodiments, the linker comprises at least one spacer unit. In some embodiments, the spacer unit or linker comprises a polyethylene glycol (PEG) moiety. In some embodiments, the PEG moiety is -(PEG) m -, where m is an integer from 1 to 10. In some embodiments, m is 2. In some other embodiments, the spacer unit or linker comprises an alkyl moiety. In some embodiments, the alkyl moiety is -(CH2) n -, wherein n is an integer from 1 to 10. In some embodiments, n is 2. In some embodiments, n is 5. In some embodiments, n is 6.

[0067] In some embodiments, the spacer unit is attached to the antibody or antigen-binding fragment via a maleimide (Mal) moiety ("Mal-spacer unit"). In some embodiments, the Mal-spacer unit is reactive with a cysteine ​​residue on the antibody or antigen-binding fragment. In some embodiments, the Mal-spacer unit is coupled to the antibody or antigen-binding fragment via a cysteine ​​residue on the antibody or antigen-binding fragment.

[0068] In some embodiments, the linker comprises a Mal-spacer unit and a cleavable peptide moiety. In some embodiments, the cleavable peptide moiety comprises an amino acid unit. In some embodiments, the cleavable peptide moiety or amino acid unit comprises Val-Cit. In some embodiments, the cleavable peptide moiety or amino acid unit comprises Val-Ala. In some embodiments, the cleavable peptide moiety or amino acid unit comprises Glu-Val-Cit. In some embodiments, the cleavable peptide moiety or amino acid unit comprises Ala-Ala-Asn. In some embodiments, the Mal-spacer unit comprises an alkyl moiety. In some embodiments, the Mal-spacer unit comprises a PEG moiety. In some embodiments, the Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the Mal-spacer unit connects the antibody or antigen-binding fragment to the cleavable portion of the linker. In some embodiments, the cleavable portion of the linker comprises a cleavable peptide moiety. In some embodiments, the cleavable peptide moiety comprises an amino acid unit. In some embodiments, the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn. In some embodiments, the linker comprises MC-Val-Cit. In some embodiments, the linker comprises MC-Val-Ala. In some embodiments, the linker comprises MC-Glu-Val-Cit. In some embodiments, the linker comprises MC-Ala-Ala-Asn. In some embodiments, the Mal-spacer unit comprises an alkyl moiety. In some embodiments, the Mal-spacer unit comprises a PEG moiety. In some embodiments, the Mal-spacer unit comprises maleimidocaproyl (MC).

[0069] In some embodiments, the cleavable portion of the linker is directly tethered to the splicing modulator, or a spacer unit connects the cleavable portion of the linker to the splicing modulator. In some embodiments, cleavage of the conjugate releases the splicing modulator from the antibody or antigen-binding fragment and the linker. In some embodiments, the spacer unit connecting the cleavable portion of the linker to the splicing modulator is self-immolative.

[0070] In some embodiments, the spacer unit connecting the cleavable portion of the linker to the splicing modulator comprises p-aminobenzyloxycarbonyl (pABC). In some embodiments, pABC connects the cleavable portion of the linker to the splicing modulator. In some embodiments, the cleavable portion of the linker comprises a cleavable peptide moiety. In some embodiments, the cleavable peptide moiety comprises an amino acid unit. In some embodiments, the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn. In some embodiments, the linker comprises Val-Cit-pABC. In some other embodiments, the linker comprises Val-Ala-pABC. In some embodiments, the linker comprises Glu-Val-Cit-pABC. In some embodiments, the linker comprises Ala-Ala-Asn-pABC.

[0071] In some embodiments, the spacer unit connecting the cleavable portion of the linker to the splicing modulator comprises p-aminobenzyl (pAB). In some embodiments, pAB connects the cleavable portion of the linker to the splicing modulator. In some embodiments, the cleavable portion of the linker comprises a cleavable peptide moiety. In some embodiments, the cleavable peptide moiety comprises an amino acid unit. In some embodiments, the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn. In some embodiments, the linker comprises Val-Cit-pAB. In some other embodiments, the linker comprises Val-Ala-pAB. In some other embodiments, the linker comprises Glu-Val-Cit-pAB. In some other embodiments, the linker comprises Ala-Ala-Asn-pAB.

[0072] In various embodiments, the linker is a non-cleavable linker. In some embodiments, the splicing modulator of the ADC is released upon degradation of the antibody or antigen-binding fragment. In some embodiments, the linker remains covalently attached to at least one amino acid of the antibody and drug upon internalization by and degradation within the target cell.

[0073] In some embodiments, the linker is a non-cleavable linker comprising at least one spacer unit. In some embodiments, the spacer unit or linker comprises a polyethylene glycol (PEG) moiety. In some embodiments, the PEG moiety is -(PEG) m -, where m is an integer from 1 to 10. In some embodiments, m is 2. In some other embodiments, the spacer unit or linker comprises an alkyl moiety. In some embodiments, the alkyl moiety is -(CH2) n -or-(CH2) n -O-(CH2) nwherein n is an integer from 1 to 10. In some embodiments, n is 2. In some embodiments, n is 5. In some embodiments, n is 6.

[0074] In some embodiments, the spacer unit of the non-cleavable linker is attached to the antibody or antigen-binding fragment via a maleimide (Mal) moiety ("Mal-spacer unit"). In some embodiments, the Mal-spacer unit is reactive with a cysteine ​​residue on the antibody or antigen-binding fragment. In some embodiments, the Mal-spacer unit is coupled to the antibody or antigen-binding fragment via a cysteine ​​residue on the antibody or antigen-binding fragment. In some embodiments, the Mal-spacer unit comprises an alkyl moiety. In some embodiments, the Mal-spacer unit comprises a PEG moiety. In some embodiments, the linker or Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the linker or Mal-spacer unit comprises maleimidocaproyl (MC) and at least one additional spacer unit. In some embodiments, the linker or Mal-spacer unit comprises MC-(PEG)2. In some embodiments, the linker or Mal-spacer unit comprises MC-(PEG)2 and at least one additional spacer unit. In some embodiments, the linker or Mal-spacer unit comprises Mal-Hex. In some embodiments, the linker or Mal-spacer unit comprises Mal-Hex and at least one additional spacer unit. In some embodiments, the linker or Mal-spacer unit comprises Mal-Et. In some embodiments, the linker or Mal-spacer unit comprises Mal-Et and at least one additional spacer unit. In some embodiments, the linker or Mal-spacer unit comprises Mal-Et-O-Et. In some embodiments, the linker or Mal-spacer unit comprises Mal-Et-O-Et and at least one additional spacer unit. In some embodiments, the Mal-spacer unit links the antibody or antigen-binding fragment to the splicing modulator.

[0075] In various embodiments, the ADC compound has formula (I): Ab-(LD) p (I) wherein Ab is an antibody or antigen-binding fragment thereof (e.g., any of the antibody or binding domain sequences disclosed herein) that targets neoplastic cells or another oncology-relevant target, such as a cancer antigen; D is any small molecule suitable for treating cancer (e.g., a splicing modulator, e.g., any of the splicing modulators disclosed herein); L is a linker (e.g., any of the linkers disclosed herein) that covalently attaches Ab to D; and p is an integer from 1 to 15. It can be represented by:

[0076] In some embodiments, the Ab is selected from any of the antibodies or binding domain sequences disclosed herein. In some embodiments, the Ab is an antibody or binding domain sequence that targets HER2 and / or neoplastic cells expressing HER2. In some embodiments, the Ab is an antibody or binding domain sequence that targets CD138 and / or neoplastic cells expressing CD138. In some embodiments, the Ab is an antibody or binding domain sequence that targets EPHA2 and / or neoplastic cells expressing EPHA2. In some embodiments, the Ab is an antibody or binding domain sequence that targets MSLN and / or neoplastic cells expressing MSLN. In some embodiments, the Ab is an antibody or binding domain sequence that targets FOLH1 and / or neoplastic cells expressing FOLH1. In some embodiments, the Ab is an antibody or binding domain sequence that targets CDH6 and / or neoplastic cells expressing CDH6. In some embodiments, the Ab is an antibody or binding domain sequence that targets CEACAM5 and / or neoplastic cells expressing CEACAM5. In some embodiments, the Ab is an antibody or binding domain sequence that targets CFClB and / or neoplastic cells expressing CFClB. In some embodiments, the Ab is an antibody or binding domain sequence that targets ENPP3 and / or neoplastic cells expressing ENPP3. In some embodiments, the Ab is an antibody or binding domain sequence that targets FOLR1 and / or neoplastic cells expressing FOLR1. In some embodiments, the Ab is an antibody or binding domain sequence that targets HAVCR1 and / or neoplastic cells expressing HAVCR1. In some embodiments, the Ab is an antibody or binding domain sequence that targets KIT and / or neoplastic cells expressing KIT. In some embodiments, the Ab is an antibody or binding domain sequence that targets MET and / or neoplastic cells expressing MET. In some embodiments, the Ab is an antibody or binding domain sequence that targets MUC16 and / or neoplastic cells expressing MUC16. In some embodiments, the Ab is an antibody or binding domain sequence that targets SLC39A6 and / or neoplastic cells expressing SLC39A6.In some embodiments, the Ab is an antibody or binding domain sequence that targets SLC44A4 and / or neoplastic cells expressing SLC44A4. In some embodiments, the Ab is an antibody or binding domain sequence that targets STEAP1 and / or neoplastic cells expressing STEAP1. In some embodiments, the Ab is an antibody or binding domain sequence that targets another cancer antigen.

[0077] In some embodiments, D is a splicing modulator. In some embodiments, D is selected from any of the splicing modulators disclosed herein. In some embodiments, D is a splicing modulator selected from D2, D1, D4, D8, D10, D11 (E7107), D20, D21, D22, D12, D25, D9, D18, D19, D13, D15, D14, D16, D17, D26, and D33, or any derivative thereof. In some embodiments, D is a splicing modulator selected from D4, D12, D15, D8, D9, D10, D13, D18, D19, D20, D21, D22, D25, and D33, or any derivative thereof. In some embodiments, D is a splicing modulator comprising D2 or any derivative thereof. In some embodiments, D is a splicing modulator comprising D1 or any derivative thereof.

[0078] In some embodiments, L is selected from any of the linkers disclosed herein or any combination of linker components disclosed herein. In some embodiments, L is a linker comprising MC-Val-Cit-pABC, Mal-(PEG)2-CO, MC-Val-Ala-pAB, MC-Val-Ala-pABC, MC-Val-Cit-pAB, Mal-Hex, Mal-Et, or Mal-Et-O-Et. In some embodiments, the linker may also comprise one or more additional spacer units. In some embodiments, L is an ADL1, ADL2, ADL5, ADL6, ADL7, ADL10, ADL12, ADL13, ADL14, ADL15, ADL21, ADL22, or ADL23 linker. In some embodiments, L is an ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15 linker. In some embodiments, L is an ADL12, ADL14, or ADL15 linker. In some embodiments, the ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15 linker may also include one or more additional spacer units. In some embodiments, L is an ADL1 linker, which may optionally include one or more additional spacer units. In some embodiments, L is an ADL2 linker, which may optionally include one or more additional spacer units. In some embodiments, L is an ADL5 linker, which may optionally include one or more additional spacer units. In some embodiments, L is an ADL6 linker, which may optionally include one or more additional spacer units. In some embodiments, L is an ADL7 linker, which may optionally include one or more additional spacer units. In some embodiments, L is an ADL12 linker, which may optionally include one or more additional spacer units. In some embodiments, L is an ADL14 linker, which may optionally include one or more additional spacer units. In some embodiments, L is an ADL15 linker, which may optionally include one or more additional spacer units.In various embodiments of the ADCs described herein, p is 1 to 10. In various embodiments, p is 2 to 8. In various embodiments, p is 4 to 8. In some embodiments, p is 4. In some embodiments, p is 8.

[0079] In some embodiments, the LD of Formula (I) is ADL1-D1. In some embodiments, the LD of Formula (I) is ADL6-D1. In some embodiments, the LD of Formula (I) is ADL5-D2. In some embodiments, the LD of Formula (I) is ADL1-D18. In some embodiments, the LD of Formula (I) is ADL5-D19. In some embodiments, the LD of Formula (I) is ADL14-D1. In some embodiments, the LD of Formula (I) is ADL12-D1. In some embodiments, the LD of Formula (I) is ADL15-D1. In some embodiments, the LD of Formula (I) is ADL12-D20. In some embodiments, the LD of Formula (I) is ADL10-D1. In some embodiments, the LD of Formula (I) is ADL12-D2. In some embodiments, the LD of Formula (I) is ADL15-D2. In some embodiments, the LD of Formula (I) is ADL12-D21. In some embodiments, the LD of Formula (I) is ADL6-D9. In some embodiments, the LD of Formula (I) is ADL1-D4. In some embodiments, the LD of Formula (I) is ADL1-D3. In some embodiments, the LD of Formula (I) is ADL1-D12. In some embodiments, the LD of Formula (I) is ADL1-D7. In some embodiments, the LD of Formula (I) is ADL1-D6. In some embodiments, the LD of Formula (I) is ADL1-D5. In some embodiments, the LD of Formula (I) is ADL22-D4. In some embodiments, the LD of Formula (I) is ADL5-D10. In some embodiments, the LD of Formula (I) is ADL5-D11. In some embodiments, the LD of Formula (I) is ADL1-D13. In some embodiments, the LD of Formula (I) is ADL1-D8. In some embodiments, the LD of Formula (I) is ADL1-D22. In some embodiments, the LD of formula (I) is ADL5-D25. In some embodiments, the LD of formula (I) is ADL12-D22. In some embodiments, the LD of formula (I) is ADL5-D15. In some embodiments, the LD of formula (I) is ADL1-D14.In some embodiments, the LD of Formula (I) is ADL5-D26. In some embodiments, the LD of Formula (I) is ADL1-D16. In some embodiments, the LD of Formula (I) is ADL5-D17. In some embodiments, the LD of Formula (I) is ADL1-D33. In some embodiments, the LD of Formula (I) is ADL1-D28. In some embodiments, the LD of Formula (I) is ADL1-D31. In some embodiments, the LD of Formula (I) is ADL1-D29. In some embodiments, the LD of Formula (I) is ADL1-D35. In some embodiments, the LD of Formula (I) is ADL5-D32. In some embodiments, the LD of Formula (I) is ADL5-D27. In some embodiments, the LD of Formula (I) is ADL12-D35. In some embodiments, the LD of Formula (I) is ADL12-D28. In some embodiments, LD of formula (I) is ADL1-D23. In some embodiments, LD of formula (I) is ADL1-D24.

[0080] In some embodiments, a pool of ADCs is provided whereby random conjugation occurs, and the average p in the pool is about 2 to about 8. In some embodiments, a pool of ADCs is provided whereby random conjugation occurs, and the average p in the pool is about 4 to about 8. In some embodiments, a pool of ADCs is provided whereby random conjugation occurs, and the average p in the pool is about 4. In some embodiments, a pool of ADCs is provided whereby random conjugation occurs, and the average p in the pool is about 8. Provided herein are compositions (e.g., pharmaceutical compositions) comprising multiple copies of any of the described ADCs, wherein the average drug loading (average p) of the ADC in the composition is about 3.5 to about 5.5 (e.g., about 4), or about 7 to about 9 (e.g., about 8).

[0081] In some embodiments, the antibody or antigen-binding fragment (Ab) of the ADC targets neoplastic cells derived from a hematological malignancy or a solid tumor. In some embodiments, the antibody or antigen-binding fragment targets neoplastic cells derived from a hematological malignancy. In some embodiments, the hematological malignancy is selected from a B-cell malignancy, leukemia (e.g., acute myeloid leukemia), lymphoma, and myeloma (e.g., multiple myeloma). In some embodiments, the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the antibody or antigen-binding fragment targets neoplastic cells derived from a solid tumor. In some embodiments, the solid tumor is selected from breast cancer (e.g., HER2-positive breast cancer), gastric cancer (e.g., gastric adenocarcinoma), prostate cancer, ovarian cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial carcinoma), salivary duct cancer, melanoma, colon cancer, cervical cancer, pancreatic cancer, renal cancer, colorectal cancer, and esophageal cancer. In some embodiments, the solid tumor is selected from HER2-positive breast cancer, gastric adenocarcinoma, prostate cancer, and osteosarcoma.

[0082] In various embodiments, the antibody or antigen-binding fragment (Ab) of the ADC is an anti-HER2 antibody or antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment binds to HER2 and targets neoplastic cells that express HER2 (i.e., the ADC targets neoplastic cells that express HER2). In some embodiments, the antibody or antigen-binding fragment of the ADC is an internalizing anti-HER2 antibody or internalizing antigen-binding fragment thereof.

[0083] In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO:1 (HCDR1), SEQ ID NO:2 (HCDR2), and SEQ ID NO:3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3). In some embodiments, the anti-HER2 antibody or antigen-binding fragment is an internalizing antibody or internalizing antigen-binding fragment. In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises human framework sequences. In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:20. In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises a human IgG heavy chain constant region. In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region. In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises a human Ig kappa or lambda light chain constant region. In some embodiments, the anti-HER2 antibody or antigen-binding fragment competes for binding to and / or binds to the same epitope as an antibody comprising a heavy chain variable domain of SEQ ID NO: 19 and a light chain variable domain of SEQ ID NO: 20.

[0084] In various embodiments, the antibody or antigen-binding fragment (Ab) of the ADC is an anti-CD138 antibody or antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment binds to CD138 and targets neoplastic cells that express CD138 (i.e., the ADC targets neoplastic cells that express CD138). In some embodiments, the antibody or antigen-binding fragment of the ADC is an internalizing anti-CD138 antibody or internalizing antigen-binding fragment thereof.

[0085] In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) 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 (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3). In some embodiments, the anti-CD138 antibody or antigen-binding fragment is an internalizing antibody or internalizing antigen-binding fragment. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises human framework sequences. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:22. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a murine IgG2a heavy chain constant region. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a murine Ig kappa light chain constant region. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a human IgG heavy chain constant region. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a human IgG2a heavy chain constant region. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a human Ig kappa or lambda light chain constant region. In some embodiments, the anti-CD138 antibody or antigen-binding fragment competes for binding to and / or binds to the same epitope as an antibody comprising a heavy chain variable domain of SEQ ID NO:21 and a light chain variable domain of SEQ ID NO:22.

[0086] In various embodiments, the antibody or antigen-binding fragment (Ab) of the ADC is an anti-EPHA2 antibody or antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment binds to EPHA2 and targets neoplastic cells that express EPHA2 (i.e., the ADC targets neoplastic cells that express EPHA2). In some embodiments, the antibody or antigen-binding fragment of the ADC is an internalizing anti-EPHA2 antibody or internalizing antigen-binding fragment thereof.

[0087] In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3). In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment is an internalizing antibody or internalizing antigen-binding fragment. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises human framework sequences. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises a human IgG heavy chain constant region. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises a human Ig kappa or lambda light chain constant region. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment competes for and / or binds to the same epitope as an antibody comprising the heavy chain variable domain of SEQ ID NO:23 and the light chain variable domain of SEQ ID NO:24.

[0088] Also provided herein, in various embodiments, are compounds comprising a linker-drug defined by the general formula: LD, where L = linker moiety, and D = drug moiety (e.g., a splicing modulator drug moiety). In various embodiments, the linker-drug (LD) compounds disclosed herein can be attached to an antibody or antigen-binding fragment and / or are suitable for use in an ADC disclosed herein, e.g., an ADC of Formula (I).

[0089] In various embodiments, the linker-drug (LD) compounds disclosed herein comprise a linker-drug structure according to formula (III): [ka] (In the formula, R 1 is selected from absent, hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 2 is absent or is a linker; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; and R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR 17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1-C6 alkyl group, -NR 15 R 16 and a linker; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR17 Selected from; R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; and Z” is [ka] Selected from; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 at least one of is hydrogen; R 2 is the linker, then R 6 or R 7 is not a linker, and R 6 or R 7 is the linker, then R 2 does not exist) or a pharmaceutically acceptable salt thereof.

[0090] In various other embodiments, the linker-drug (LD) compounds disclosed herein comprise a linker-drug structure according to formula (V): [ka] (In the formula, R 1 is selected from absent, hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 2 is absent or is a linker; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; and R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR 17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1-C6 alkyl group, -NR 15 R 16 and a linker; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 selected from; and R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; R 1 , R 2 , R 3, R 4 , R 5 , R 6 , R 7 , R 8 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 at least one of is hydrogen; R 2 is the linker, then R 6 or R 7 is not a linker, and R 6 or R 7 is the linker, then R 2 does not exist) or a pharmaceutically acceptable salt thereof.

[0091] In various other embodiments, the linker-drug (LD) compounds disclosed herein comprise a linker-drug structure according to formula (VII): In various embodiments, the present disclosure provides compounds of formula (VII): [ka] (In the formula, R 1 and R 9 are each independently selected from absent, hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 2 is absent or is a linker; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR 17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1-C6 alkyl group, -NR 15 R 16 and a linker; R 10 is selected from hydrogen, a C1-C6 alkyl group, a -C(=O)-(C1-C6 alkyl) group, and -CD3; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 Selected from; R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; and a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 15 , R16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 at least one of is hydrogen; R 2 is the linker, then R 6 or R 7 is not a linker, and R 6 or R 7 is the linker, then R 2 does not exist; R 1 and R 9 (It is impossible for both to exist) or a pharmaceutically acceptable salt thereof.

[0092] In various other embodiments, the linker-drug (LD) compounds disclosed herein comprise a linker-drug structure according to formula (IX): [ka] (In the formula, R 1 is selected from absent, hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 2 is a linker; R 3is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; R 4 is selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 10 is selected from a 3- to 10-membered carbocyclic ring and a 3- to 10-membered heterocyclic ring, each of which is selected from 0 to 3 R a Each R is replaced by a are independently a halogen, a C1-C6 alkyl group, an -O-(C1-C6) alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylhydroxy group, or -S(=O) w -(4- to 7-membered heterocycle), a 4- to 7-membered carbocycle, and a 4- to 7-membered heterocycle; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 selected from; and R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; R 1 , R 2 , R 3 , R 4 , R 10 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; Each Ra are independently a halogen, a hydroxyl group, or -NR 15 R 16 , C1-C6 alkyl group, -(C=O)-(C1-C6 alkyl) group, -(C=O)-(C1-C6 alkyl)-(C3-C 10 heterocyclyl) groups, and C1-C6 alkylcarboxylic acid groups (wherein each of these is selected from the group consisting of halogen, hydroxyl, -NR 15 R 16 and C1-C3 alkyl; and w is 0, 1, or 2) or a pharmaceutically acceptable salt thereof.

[0093] Also provided herein, in various embodiments, are therapeutic uses for the described ADC compounds and compositions, e.g., in the treatment of neoplastic disorders, e.g., cancer. In certain aspects, the disclosure provides methods of treating neoplastic disorders, e.g., cancer, that express an antigen targeted by the antibody or antigen-binding fragment of the ADC, such as HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFClB, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, or STEAP1.

[0094] In certain aspects, the disclosure provides methods of treating a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount and / or therapeutically effective regimen of any one of the described ADCs or compositions. In some embodiments, the neoplastic disorder is a hematological malignancy or a solid tumor. In some embodiments, the neoplastic disorder is a hematological malignancy. In some embodiments, the hematological malignancy is selected from a B-cell malignancy, leukemia, lymphoma, and myeloma. In some embodiments, the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the neoplastic disorder is a solid tumor. In some embodiments, the solid tumor is selected from breast cancer (e.g., HER2-positive breast cancer), gastric cancer (e.g., gastric adenocarcinoma), prostate cancer, ovarian cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial cancer), salivary duct cancer, melanoma, colon cancer, cervical cancer, pancreatic cancer, renal cancer, colorectal cancer, and esophageal cancer. In some embodiments, the solid tumor is selected from HER2-positive breast cancer, gastric adenocarcinoma, prostate cancer, and osteosarcoma.

[0095] In some embodiments, treatment with the antibody-drug conjugate or composition induces bystander killing of neoplastic cells that do not express the target antigen but are adjacent to neoplastic cells that do express the target antigen, hi some embodiments, a subject has one or more neoplastic cells that express the target antigen.

[0096] In some embodiments, the target antigen is HER2. In some embodiments, the one or more neoplastic cells are derived from HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial carcinoma), osteosarcoma, or salivary duct cancer. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-HER2 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0097] In some embodiments, the target antigen is CD138. In some embodiments, the one or more neoplastic cells are derived from multiple myeloma that expresses CD138. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-CD138 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0098] In some embodiments, the target antigen is EPHA2. In some embodiments, the one or more neoplastic cells are derived from EPHA2-expressing breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer. In some embodiments, the subject is refractory or poorly responsive to treatment with (a) an anti-EPHA2 antibody when administered alone and / or (b) a splicing regulator when administered alone. In some embodiments, the subject is intolerant, refractory, or poorly responsive to treatment with a splicing regulator when administered alone.

[0099] In some embodiments, the target antigen is MSLN. In some embodiments, the one or more neoplastic cells are derived from ovarian cancer, cervical cancer, pancreatic cancer, or lung cancer (e.g., lung adenocarcinoma) that expresses MSLN. In some embodiments, the subject is refractory or unresponsive to treatment with (a) an anti-MSLN antibody administered alone and / or (b) a splicing modulator administered alone. In some embodiments, the subject is intolerant, unresponsive, or unresponsive to treatment with a splicing modulator administered alone.

[0100] In some embodiments, the target antigen is FOLH1. In some embodiments, the one or more neoplastic cells are derived from a prostate cancer that expresses FOLH1. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-FOLH1 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0101] In some embodiments, the target antigen is CDH6. In some embodiments, the one or more neoplastic cells are derived from a CDH6-expressing renal carcinoma. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-CDH6 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0102] In some embodiments, the target antigen is CEACAM5. In some embodiments, the one or more neoplastic cells are derived from a colorectal cancer that expresses CEACAM5. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-CEACAM5 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0103] In some embodiments, the target antigen is CFC1B. In some embodiments, the one or more neoplastic cells are derived from a pancreatic cancer that expresses CFC1B. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-CFC1B antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0104] In some embodiments, the target antigen is ENPP3. In some embodiments, the one or more neoplastic cells are derived from a renal cancer that expresses ENPP3. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-ENPP3 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0105] In some embodiments, the target antigen is FOLR1. In some embodiments, the one or more neoplastic cells are derived from an ovarian cancer that expresses FOLR1. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-FOLR1 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0106] In some embodiments, the target antigen is HAVCR1. In some embodiments, the one or more neoplastic cells are derived from a renal cancer or an esophageal cancer that expresses HAVCR1. In some embodiments, the subject is refractory or poorly responsive to treatment with (a) an anti-HAVCR1 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, refractory, or poorly responsive to treatment with a splicing modulator when administered alone.

[0107] In some embodiments, the target antigen is KIT. In some embodiments, the one or more neoplastic cells are derived from a KIT-expressing renal carcinoma. In some embodiments, the subject is refractory or unresponsive to treatment with (a) an anti-KIT antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, unresponsive, or unresponsive to treatment with a splicing modulator when administered alone.

[0108] In some embodiments, the target antigen is MET. In some embodiments, the one or more neoplastic cells are derived from MET-expressing renal or esophageal cancer. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-MET antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0109] In some embodiments, the target antigen is MUC16. In some embodiments, the one or more neoplastic cells are derived from ovarian, cervical, or breast cancer that expresses MUC16. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-MUC16 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0110] In some embodiments, the target antigen is SLC39A6. In some embodiments, the one or more neoplastic cells are derived from breast cancer or prostate cancer that expresses SLC39A6. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-SLC39A6 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0111] In some embodiments, the target antigen is SLC44A4. In some embodiments, the one or more neoplastic cells are derived from a prostate cancer that expresses SLC44A4. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-SLC44A4 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0112] In some embodiments, the target antigen is STEAP1. In some embodiments, the one or more neoplastic cells are derived from a prostate cancer that expresses STEAP1. In some embodiments, the subject is refractory or non-responsive to treatment with (a) an anti-STEAP1 antibody when administered alone and / or (b) a splicing modulator when administered alone. In some embodiments, the subject is intolerant, non-responsive, or non-responsive to treatment with a splicing modulator when administered alone.

[0113] In certain other aspects, the disclosure provides methods of reducing or inhibiting tumor growth in a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount and / or therapeutically effective regimen of any one of the described ADCs or compositions.

[0114] In some embodiments, treatment with the antibody-drug conjugate or composition induces bystander killing of neoplastic tumor cells that do not express the target antigen but that are adjacent to neoplastic tumor cells that do express the target antigen, hi some embodiments, a tumor comprises one or more neoplastic cells that express the target antigen.

[0115] In some embodiments, the target antigen is HER2. In some embodiments, the one or more neoplastic cells are derived from HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial carcinoma), osteosarcoma, or salivary duct cancer. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-HER2 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0116] In some embodiments, the target antigen is CD138. In some embodiments, the one or more neoplastic cells are derived from multiple myeloma that expresses CD138. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-CD138 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0117] In some embodiments, the target antigen is EPHA2. In some embodiments, the one or more neoplastic cells are derived from EPHA2-expressing breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-EPHA2 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0118] In some embodiments, the target antigen is MSLN. In some embodiments, the one or more neoplastic cells are derived from MSLN-expressing ovarian cancer, cervical cancer, pancreatic cancer, or lung cancer (e.g., lung adenocarcinoma). In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-MSLN antibody administered alone and / or (b) a splicing modulator administered alone.

[0119] In some embodiments, the target antigen is FOLH1. In some embodiments, the one or more neoplastic cells are derived from a prostate cancer that expresses FOLH1. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-FOLH1 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0120] In some embodiments, the target antigen is CDH6. In some embodiments, the one or more neoplastic cells are derived from a CDH6-expressing renal carcinoma. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-CDH6 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0121] In some embodiments, the target antigen is CEACAM5. In some embodiments, the one or more neoplastic cells are derived from a colorectal cancer that expresses CEACAM5. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-CEACAM5 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0122] In some embodiments, the target antigen is CFC1B. In some embodiments, the one or more neoplastic cells are derived from a pancreatic cancer that expresses CFC1B. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-CFC1B antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0123] In some embodiments, the target antigen is ENPP3. In some embodiments, the one or more neoplastic cells are derived from a renal carcinoma that expresses ENPP3. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-ENPP3 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0124] In some embodiments, the target antigen is FOLR1. In some embodiments, the one or more neoplastic cells are derived from an ovarian cancer that expresses FOLR1. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-FOLR1 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0125] In some embodiments, the target antigen is HAVCR1. In some embodiments, the one or more neoplastic cells are derived from a renal cancer or an esophageal cancer that expresses HAVCR1. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-HAVCR1 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0126] In some embodiments, the target antigen is KIT. In some embodiments, the one or more neoplastic cells are derived from a KIT-expressing renal carcinoma. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-KIT antibody administered alone and / or (b) a splicing modulator administered alone.

[0127] In some embodiments, the target antigen is MET. In some embodiments, the one or more neoplastic cells are derived from a MET-expressing renal or esophageal cancer. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-MET antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0128] In some embodiments, the target antigen is MUC16. In some embodiments, the one or more neoplastic cells are derived from ovarian, cervical, or breast cancer that expresses MUC16. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-MUC16 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0129] In some embodiments, the target antigen is SLC39A6. In some embodiments, the one or more neoplastic cells are derived from breast or prostate cancer that expresses SLC39A6. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-SLC39A6 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0130] In some embodiments, the target antigen is SLC44A4. In some embodiments, the one or more neoplastic cells are derived from a prostate cancer that expresses SLC44A4. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-SLC44A4 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0131] In some embodiments, the target antigen is STEAP1. In some embodiments, the one or more neoplastic cells are derived from a prostate cancer that expresses STEAP1. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-STEAP1 antibody when administered alone and / or (b) a splicing modulator when administered alone.

[0132] In yet another aspect, the disclosure provides methods for determining whether a subject having or suspected of having a neoplastic disorder will respond to treatment with any one of the described ADCs or compositions by providing a biological sample from the subject and contacting the biological sample with the ADC or composition. In some embodiments, the biological sample is a tumor sample. In some embodiments, the tumor sample is a tumor biopsy or a blood sample. In some embodiments, the blood sample is selected from blood, a blood fraction, or cells obtained from blood or a blood fraction. In some embodiments, the subject has one or more neoplastic cells that express a target antigen. In some embodiments, the target antigen is HER2. In some embodiments, the one or more neoplastic cells are derived from HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial carcinoma), osteosarcoma, or salivary duct cancer. In some embodiments, the target antigen is CD138. In some embodiments, the one or more neoplastic cells are derived from CD138-expressing multiple myeloma. In some embodiments, the target antigen is EPHA2. In some embodiments, the one or more neoplastic cells are derived from breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer that expresses EPHA2. In some embodiments, the target antigen is MSLN. In some embodiments, the one or more neoplastic cells are derived from ovarian cancer, cervical cancer, pancreatic cancer, or lung cancer (e.g., lung adenocarcinoma) that expresses MSLN. In some embodiments, the target antigen is FOLH1. In some embodiments, the one or more neoplastic cells are derived from prostate cancer that expresses FOLH1. In some embodiments, the target antigen is CDH6. In some embodiments, the one or more neoplastic cells are derived from renal cancer that expresses CDH6. In some embodiments, the target antigen is CEACAM5. In some embodiments, the one or more neoplastic cells are derived from colorectal cancer that expresses CEACAM5. In some embodiments, the target antigen is CFC1B. In some embodiments, the one or more neoplastic cells are derived from pancreatic cancer that expresses CFC1B. In some embodiments, the target antigen is ENPP3. In some embodiments, the one or more neoplastic cells are derived from a renal carcinoma that expresses ENPP3.In some embodiments, the target antigen is FOLR1. In some embodiments, the one or more neoplastic cells are derived from an ovarian cancer that expresses FOLR1. In some embodiments, the target antigen is HAVCR1. In some embodiments, the one or more neoplastic cells are derived from a renal cancer or esophageal cancer that expresses HAVCR1. In some embodiments, the target antigen is KIT. In some embodiments, the one or more neoplastic cells are derived from a renal cancer that expresses KIT. In some embodiments, the target antigen is MET. In some embodiments, the one or more neoplastic cells are derived from a renal cancer or esophageal cancer that expresses MET. In some embodiments, the target antigen is MUC16. In some embodiments, the one or more neoplastic cells are derived from an ovarian cancer, cervical cancer, or breast cancer that expresses MUC16. In some embodiments, the target antigen is SLC39A6. In some embodiments, the one or more neoplastic cells are derived from a breast cancer or prostate cancer that expresses SLC39A6. In some embodiments, the target antigen is SLC44A4. In some embodiments, the one or more neoplastic cells are derived from a prostate cancer that expresses SLC44A4. In some embodiments, the target antigen is STEAP 1. In some embodiments, the one or more neoplastic cells are derived from a prostate cancer that expresses STEAP 1.

[0133] Further provided herein, in various embodiments, are pharmaceutical compositions comprising an ADC and a pharmaceutically acceptable diluent, carrier, and / or excipient. Methods of making the described ADC compounds and compositions are also disclosed. [Brief explanation of the drawings]

[0134] [Figure 1]Figure 1 shows the dose response of exemplary payload compounds in a competitive binding assay. Nuclear extracts from 293F cells overexpressing wild-type flag-tagged SF3B1 were immunoprecipitated with anti-SF3B1 antibody and a scintillation proximity assay (SPA) bead cocktail. Binding reactions included the antibody-bead mixture and increasing concentrations of compound, followed by competition with a 3H-labeled pladienolide B (PB) probe. The y-axis represents the percent change in specific binding (% response) compared to the DMSO control (0%). Data are presented as mean ± standard deviation (SD). [Figure 2] Figure 1 shows modulation of splicing by exemplary payload compounds in an in vitro splicing assay. Nuclear extracts from HeLa S3 cells were incubated with Ad2.2 pre-mRNA and increasing concentrations of compounds, and splicing modulation was subsequently quantified by RT-PCR. The Ad2.2 sequence is derived from an adenoviral Ad2 pre-mRNA substrate with modifications around the branch site sequence. The y-axis represents the percent change in splicing (% response) compared to the DMSO control (0%). Data are presented as mean ± SD. [Figure 3]

[0023] Figure 1 shows the viability dose response of exemplary payload compounds in HER2-amplified breast cancer cells (HCC1954). Cells were incubated with compounds for 144 hours (6 days) and viability was read using CellTiter-Glo® reagent. Data are presented as mean ± SD. [Figure 4]

[0023] Figure 1 shows the results of a cell binding assay. Binding of exemplary HER2-ADCs to JIMT1 cells was determined by flow cytometry. Mean fluorescence intensity values ​​were measured to determine binding of the conjugate followed by the PE-labeled secondary. Data are presented as mean values ​​± SD. [Figure 5A] FIG. 5A shows the viability dose response of an exemplary HER2-ADC in HER2-amplified breast cancer cells (HCC1954). [Figure 5B] Figure 5B shows the viability dose response of an exemplary HER2-ADC in HER2-amplified gastric cancer cells (N87). [Figure 5C]Cells were incubated with the conjugates for 144 hours (6 days) and viability was read with CellTiter-Glo® reagent. Data are presented as mean ± SD. [Figure 6]

[0023] Figure 1 shows the viability dose response of an exemplary HER2-ADC in non-HER2-expressing breast cancer cells (MCF7). Cells were incubated with the conjugate for 144 hours (6 days) and viability was read with CellTiter-Glo® reagent. Data are presented as mean ± SD. [Figure 7] Figure 1 shows the results of an SLC25A19 splicing assay in HER2-amplified breast cancer cells (HCC1954). Cells were incubated with the conjugate for 24 hours, and splicing of the SLC25A19 transcript was measured in real-time qPCR reactions using a specific Taqman primer-probe set. The y-axis represents the percent (%) response compared to the DMSO control (0%). Data are presented as mean ± SD. [Figure 8]

[0023] Figure 1 shows the results of a bystander killing assay. HER2-overexpressing H1568 cells (target positive) or luciferase-tagged H1568 cells (target negative) were treated with exemplary HER2-ADCs, either plated alone or incubated together in co-culture for 144 hours (6 days). Plates were read with OneGlo® reagent. The y-axis represents the percent (%) response compared to the PBS control (100%). Data are presented as mean ± SD. [Figure 9] Figure 1 shows tumor growth kinetics for each group of HCC1954-implanted CB17-SCID mice (6-10 animals per group) treated with a single intravenous dose of an exemplary HER2-ADC or the corresponding dose-matched payload. Tumor volumes were measured twice weekly after treatment. Data are presented as mean ± standard error of the mean (SEM). [Figure 10]

[0023] Figure 1 shows the viability dose response of an exemplary CD138-ADC in CD138-expressing multiple myeloma cell lines. MOLP8 cells were incubated with the conjugate for 144 hours (6 days) and viability was read with CellTiter-Glo® reagent. Data are presented as mean ± SD. [Figure 11]

[0023] Figure 1 shows the viability dose response of an exemplary EPH2A-ADC in EPHA2-expressing prostate cancer cell lines. PC3 cells were incubated with the conjugate for 144 hours (6 days) and viability was read with CellTiter-Glo® reagent. Data are presented as mean ± SD. [Figure 12A] 12A and 12B show the results of an in vitro stability assay for an exemplary anti-HER2 ADC, AB185-ADL1-D1. The y-axis represents the concentration of total antibody (FIG. 12A) and conjugated (intact) payload (FIG. 12B); the x-axis represents time as measured in hours at 37° C. [Figure 12B] 12A and 12B show the results of an in vitro stability assay for an exemplary anti-HER2 ADC, AB185-ADL1-D1. The y-axis represents the concentration of total antibody (FIG. 12A) and conjugated (intact) payload (FIG. 12B); the x-axis represents time as measured in hours at 37° C. [Figure 13] 1 shows plasma concentrations of an exemplary anti-HER2 ADC, AB185-ADL1-D1, after a single intravenous administration in CD17-SCID N87 tumor-bearing mice. [Figure 14] 1 shows a schematic diagram of an exemplary RNA sequencing and protein ligandome experiment. [Figure 15] 1 shows a schematic diagram of an exemplary T cell priming experiment. [Figure 16] The results of FACS analysis are shown. Monocytes were isolated from peripheral blood mononuclear cells (PBMCs) and induced to differentiate into dendritic cells (DCs) by culturing them in a cytokine cocktail. FACS was performed to confirm the maturation of DCs from monocytes. [Figure 17A]Figure 17 shows the results of an ELISpot assay. Figure 17A shows an ELISpot plate indicating priming of CD8+ T cell activation by neoantigen 1. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17B shows quantification of IFNγ spots (number of spots) in the neoantigen 1 ELISpot plate (Figure 17A). Figure 17C shows an ELISpot plate indicating priming of CD8+ T cell activation by neoantigen 3. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17D shows quantification of IFNγ spots (fold change) in the neoantigen 3 ELISpot plate (Figure 17C). [Figure 17B] Figure 17 shows the results of an ELISpot assay. Figure 17A shows an ELISpot plate indicating priming of CD8+ T cell activation by neoantigen 1. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17B shows quantification of IFNγ spots (number of spots) in the neoantigen 1 ELISpot plate (Figure 17A). Figure 17C shows an ELISpot plate indicating priming of CD8+ T cell activation by neoantigen 3. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17D shows quantification of IFNγ spots (fold change) in the neoantigen 3 ELISpot plate (Figure 17C). [Figure 17C] Figure 17 shows the results of an ELISpot assay. Figure 17A shows an ELISpot plate indicating priming of CD8+ T cell activation by neoantigen 1. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17B shows quantification of IFNγ spots (number of spots) in the neoantigen 1 ELISpot plate (Figure 17A). Figure 17C shows an ELISpot plate indicating priming of CD8+ T cell activation by neoantigen 3. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17D shows quantification of IFNγ spots (fold change) in the neoantigen 3 ELISpot plate (Figure 17C). [Figure 17D]Figure 17 shows the results of an ELISpot assay. Figure 17A shows an ELISpot plate indicating priming of CD8+ T cell activation by neoantigen 1. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17B shows quantification of IFNγ spots (number of spots) in the neoantigen 1 ELISpot plate (Figure 17A). Figure 17C shows an ELISpot plate indicating priming of CD8+ T cell activation by neoantigen 3. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17D shows quantification of IFNγ spots (fold change) in the neoantigen 3 ELISpot plate (Figure 17C). [Figure 18] 1 shows a plot comparing splicing efficacy (IC50 qPCR) to cellular efficacy (GI50 CTG) for exemplary anti-HER2 ADCs in HCC1954 breast cancer cells. Values ​​scale according to cell lethality and are shaded according to the depth of the alternative splicing reaction. [Figure 19] 1 shows a plot comparing splicing efficacy (IC50 qPCR) to cellular efficacy (GI50 CTG) for exemplary anti-HER2 ADCs in N87 gastric cancer cells. Values ​​scale according to cell lethality and are shaded according to the depth of the alternative splicing reaction. [Figure 20] 1 shows plots comparing the efficacy and lethality (in HCC1954 breast cancer cells) of exemplary anti-HER2 ADCs with the stability and permeability of the corresponding payloads. Values ​​are scaled according to payload stability and shaded according to payload permeability. [Figure 21] Figure 1 shows tumor growth kinetics for groups of N87-implanted CB17-SCID mice (N=8 per group) treated intravenously with vehicle or 10 mg / kg trastuzumab, TDM1, or exemplary HER2-ADC Q7D for two cycles. Data are presented as mean ± standard error of the mean (SEM) (mm). [Figure 22]Figure 1 shows the weight change for each group of N87-implanted CB17-SCID mice (N=8 per group) treated intravenously with vehicle or 10 mg / kg trastuzumab, TDM1, or exemplary HER2-ADC Q7D for two cycles. Data are expressed as the mean ± standard error of the mean (SEM) (%). [Figure 23] Tumor growth kinetics (left) and body weight change (right) are shown for groups of N87-implanted CB17-SCID mice (N=8 per group) treated intravenously with vehicle or 10 mg / kg trastuzumab, TDM1, or exemplary HER2-ADC Q7D for two cycles. Data are expressed as mean ± SEM (tumor volume, mm) or mean ± SEM (body weight, %). [Figure 24A] Figure 24 shows pharmacodynamic (PD) modulation of mRNA junctions in N87-implanted CB17-SCID mice treated intravenously with vehicle or 10 mg / kg trastuzumab, TDM1, or the exemplary HER2-ADC Q7D for two cycles. RT-qPCR of FBXW5 (mature mRNA transcript) was monitored, as shown in Figures 24A and 24C. RT-qPCR of TAOK1 (neojunction transcript) was monitored, as shown in Figures 24B and 24D. Animals (N=4 per group) were harvested at either 48 hours (Figures 24A and 24B) or the indicated times (Figures 24C and 24D). Tumors were excised for RNA extraction and RT-qPCR. [Figure 24B] Figure 24 shows pharmacodynamic (PD) modulation of mRNA junctions in N87-implanted CB17-SCID mice treated intravenously with vehicle or 10 mg / kg trastuzumab, TDM1, or the exemplary HER2-ADC Q7D for two cycles. RT-qPCR of FBXW5 (mature mRNA transcript) was monitored, as shown in Figures 24A and 24C. RT-qPCR of TAOK1 (neojunction transcript) was monitored, as shown in Figures 24B and 24D. Animals (N=4 per group) were harvested at either 48 hours (Figures 24A and 24B) or the indicated times (Figures 24C and 24D). Tumors were excised for RNA extraction and RT-qPCR. [Figure 24C]Figure 24 shows pharmacodynamic (PD) modulation of mRNA junctions in N87-implanted CB17-SCID mice treated intravenously with vehicle or 10 mg / kg trastuzumab, TDM1, or the exemplary HER2-ADC Q7D for two cycles. RT-qPCR of FBXW5 (mature mRNA transcript) was monitored, as shown in Figures 24A and 24C. RT-qPCR of TAOK1 (neojunction transcript) was monitored, as shown in Figures 24B and 24D. Animals (N=4 per group) were harvested at either 48 hours (Figures 24A and 24B) or the indicated times (Figures 24C and 24D). Tumors were excised for RNA extraction and RT-qPCR. [Figure 24D] Figure 24 shows pharmacodynamic (PD) modulation of mRNA junctions in N87-implanted CB17-SCID mice treated intravenously with vehicle or 10 mg / kg trastuzumab, TDM1, or the exemplary HER2-ADC Q7D for two cycles. RT-qPCR of FBXW5 (mature mRNA transcript) was monitored, as shown in Figures 24A and 24C. RT-qPCR of TAOK1 (neojunction transcript) was monitored, as shown in Figures 24B and 24D. Animals (N=4 per group) were harvested at either 48 hours (Figures 24A and 24B) or the indicated times (Figures 24C and 24D). Tumors were excised for RNA extraction and RT-qPCR. [Figure 25] FIG. 1 shows a schematic diagram of an exemplary target adaptive disease analysis. [Figure 26] 1 shows an exemplary bioconjugation scheme for the preparation of ADCs using splicing modulators. DETAILED DESCRIPTION OF THE INVENTION

[0135] The disclosed compositions and methods may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures, which form a part of this disclosure.

[0136] Throughout this document, descriptions refer to compositions and methods of using the compositions. Where this disclosure describes or claims features or embodiments related to a composition, such features or embodiments are equally applicable to methods of using the composition. Similarly, where this disclosure describes or claims features or embodiments related to methods of using a composition, such features or embodiments are equally applicable to the composition.

[0137] When a range of values ​​is expressed, it includes embodiments using any specific value within that range. Furthermore, reference to values ​​stated in ranges includes each individual value within that range. All ranges are inclusive of their endpoints and are combinable. When values ​​are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another embodiment. Reference to a particular numerical value includes at least the particular value, unless the context clearly dictates otherwise. The use of "or" is intended to mean "and / or" unless the specific context in which it is used dictates otherwise. All references cited herein are incorporated by reference for all purposes. In the event of a conflict between a reference and the present specification, the present specification shall control.

[0138] It should be understood that certain features of the compositions and methods of the present disclosure, which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the compositions and methods of the present disclosure, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0139] definition Throughout this specification and claims, various terms are used with respect to the described embodiments. Such terms should be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms should be construed consistent with the definition provided herein.

[0140] As used herein, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise.

[0141] The term "about" or "approximately," in connection with numerical values ​​and ranges, refers to a value or range that approximates or is close to the stated value or range such that the embodiment will operate as intended, such as having a desired amount of nucleic acid or polypeptide in a reaction mixture, as will be apparent to one of ordinary skill in the art from the teachings contained herein. In some embodiments, about means ±10% of a numerical amount.

[0142] The terms "antibody-drug conjugate," "antibody conjugate," "conjugate," "immunoconjugate," and "ADC" are used interchangeably and represent the general formula: Ab-(LD) p"Ab" refers to one or more therapeutic compounds (e.g., splicing modulators) linked to one or more antibodies or antigen-binding fragments defined by Formula I, where Ab = antibody or antigen-binding fragment, L = linker moiety, D = drug moiety (e.g., splicing modulator drug moiety), and p = number of drug moieties per antibody or antigen-binding fragment. ADCs comprising splicing modulator drug moieties may also be more specifically referred to herein as "splicing modulator-loaded antibodies" or "SMLAs." In ADCs comprising splicing modulator drug moieties, "p" refers to the number of splicing modulator compounds linked to an antibody or antigen-binding fragment. In some embodiments, the linker L may comprise a cleavable moiety between the antibody or antigen-binding fragment and the therapeutic compound. In some embodiments, the linker L may comprise a cleavable moiety that can be attached to either or both of the antibody or antigen-binding fragment and the therapeutic compound via one or more spacer units. In some embodiments, when a spacer unit links a cleavable moiety to a therapeutic compound, it is a self-immolative spacer unit. In other embodiments, the linker L does not include a cleavable moiety and is a non-cleavable linker. In some embodiments, the linker L may include at least one spacer unit that can be directly attached to an antibody or antigen-binding fragment and to a therapeutic compound. Exemplary cleavable and non-cleavable linkers are described and exemplified herein.

[0143] The term "antibody" is used in its 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. The heavy chain of an antibody is composed of a heavy chain variable domain (V H ) and heavy chain constant region (C H The light chain consists of a light chain variable domain (V L ) and the light chain constant domain (C L). For purposes of this application, mature heavy and light chain variable domains each comprise three complementarity-determining regions (CDR1, CDR2, and CDR3) within four framework regions (FR1, FR2, FR3, and FR4), flanked from N-terminus to C-terminus by FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. An "antibody" can be naturally occurring or artificial, such as a monoclonal antibody produced by conventional hybridoma technology. The term "antibody" includes full-length monoclonal and polyclonal antibodies, as well as antibody fragments such as Fab, Fab', F(ab')2, Fv, and single-chain antibodies. An antibody can be any one of the five major immunoglobulin classes: IgA, IgD, IgE, IgG, and IgM, or any subclass thereof (e.g., isotypes IgG1, IgG2, IgG3, IgG4). The term also encompasses human antibodies, chimeric antibodies, humanized antibodies, and any modified immunoglobulin molecule containing an antigen recognition site, so long as it exhibits the desired biological activity (e.g., binding to a target antigen, internalizing into a target antigen-expressing cell).

[0144] The term "monoclonal antibody," as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies; i.e., the individual antibodies comprising the population are identical except for possible minor naturally occurring mutations. Monoclonal antibodies are highly specific, directed against a single antigenic epitope. In contrast, conventional (polyclonal) antibody preparations typically include a large number of antibodies directed against (or specific for) different epitopes. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous antibody population and should not be construed as requiring that the antibody have been produced by any particular method. For example, monoclonal antibodies to be used in the present disclosure may be produced by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or may be produced by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). Monoclonal antibodies may also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature 352:624-8 and Marks et al. (1991) J Mol Biol. 222:581-97, for example.

[0145] The monoclonal antibodies described herein specifically include "chimeric" antibodies in which a portion of the heavy and / or light chain is identical to or homologous to corresponding sequences in antibodies from a particular species or belonging to a particular antibody class or subclass, while the remainder of one or more chains is identical to or homologous to corresponding sequences in antibodies 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.

[0146] 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.

[0147] The term "chimeric antibody," as used herein, refers to an antibody in which the amino acid sequences of the immunoglobulin molecules are derived from two or more species. In some instances, the variable regions of both the heavy and light chains correspond to the variable regions of antibodies derived from one species having the desired specificity, affinity, and activity, while the constant regions are homologous to antibodies derived from another species (e.g., human), thereby minimizing the immune response in the latter species.

[0148] As used herein, the term "humanized antibody" refers to forms of antibodies that contain sequences from non-human (e.g., murine) antibodies as well as human antibodies. Such antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulins. Generally, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the framework (FR) regions are those of a human immunoglobulin. A humanized antibody will optionally also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. Humanized antibodies can be further modified by substitution of residues within either the Fv framework regions and / or replaced non-human residues to refine and optimize antibody specificity, affinity, and / or activity.

[0149] The term "antigen-binding fragment" or "antigen-binding portion" of an antibody, as used herein, refers to one or more fragments of an antibody or protein that retain the ability to specifically bind to an antigen (e.g., HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, STEAP1). Antigen-binding fragments may also retain the ability to internalize into antigen-expressing cells. In some embodiments, antigen-binding fragments also retain immune effector activity. It has been shown that fragments of full-length antibodies can perform the antigen-binding function of the full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding fragment" or "antigen-binding portion" of an antibody include (i) V L , V H , C L , and C H1 (ii) a F(ab')2 fragment, a bivalent fragment containing two Fab fragments linked by a disulfide bridge at the hinge region; and (iii) a V H and C H1 (iv) a single-arm V fragment of an antibody; L and V H (v) a single variable domain, e.g., a V H and (vi) isolated complementarity-determining regions (CDRs). In addition, two domains of an Fv fragment, V, V, and VD, are included. L and V H are encoded by separate genes, but using recombinant methods they can be joined together with synthetic linkers to form V L and V HThese regions can be paired to form a single protein chain (known as a single-chain Fv (scFv)) that forms a monovalent molecule. See, e.g., Bird et al. (1988) Science 242:423-6; and Huston et al. (1988) Proc Natl Acad Sci. USA 85:5879-83. Such single-chain antibodies are also intended to be encompassed within the term "antigen-binding fragment" or "antigen-binding portion" of an antibody and are known in the art as an exemplary type of binding fragment that can be internalized into cells upon binding (see, e.g., Zhu et al. (2010) 9:2131-41; He et al. (2010) J Nucl Med. 51:427-32; and Fitting et al. (2015) MAbs 7:390-402). In certain embodiments, scFv molecules can be incorporated into fusion proteins. Other forms of single chain antibodies are also encompassed, such as diabodies. H and V L A bivalent, bispecific antibody is one in which the domains are expressed on a single polypeptide chain, but use a linker that is short enough to not allow pairing between the two domains on the same chain, thereby forcing them to pair with complementary domains on another chain to create two antigen-binding sites (see, e.g., Holliger et al. (1993) Proc Natl Acad Sci. USA 90:6444-8; and Poljak et al. (1994) Structure 2:1121-3). Antigen-binding fragments are obtained using conventional techniques known to those skilled in the art, and such binding fragments are screened for utility (e.g., binding affinity, internalization) in the same manner as intact antibodies. Antigen-binding fragments can be prepared by cleaving the intact protein, for example, by protease or chemical cleavage.

[0150] "Internalizing," as used herein in reference to an antibody or antigen-binding fragment, refers to an antibody or antigen-binding fragment that, upon binding to a cell, has the ability to penetrate the lipid bilayer membrane of the cell and be incorporated into an internal compartment of the cell, preferably a degradable compartment (i.e., be "internalized"). For example, an internalizing anti-HER2 antibody is one that has the ability to be incorporated into the cell after binding to HER2 on the cell membrane. In some embodiments, the antibody or antigen-binding fragment used in the ADCs disclosed herein targets a cell surface antigen (e.g., HER2) and is an internalizing antibody or internalizing antigen-binding fragment (i.e., the ADC translocates through the cell membrane after antigen binding). In some embodiments, the internalizing antibody or antigen-binding fragment binds to a receptor on the cell surface. An internalizing antibody or internalizing antigen-binding fragment that targets a receptor on the cell membrane can induce receptor-mediated endocytosis. In some embodiments, the internalizing antibody or internalizing antigen-binding fragment is incorporated into the cell by receptor-mediated endocytosis.

[0151] "Non-internalizing," as used herein in reference to an antibody or antigen-binding fragment, refers to an antibody or antigen-binding fragment that remains on the cell surface upon binding to a cell. In some embodiments, the antibody or antigen-binding fragment used in the ADCs disclosed herein targets a cell surface antigen and is a non-internalizing antibody or non-internalizing antigen-binding fragment (i.e., the ADC remains on the cell surface after antigen binding and does not translocate through the cell membrane). In some embodiments, the non-internalizing antibody or antigen-binding fragment binds to a non-internalizing receptor or other cell surface antigen. Exemplary non-internalizing cell surface antigens include, but are not limited to, CA125 and CEA, and antibodies that bind to non-internalizing antigen targets are also known in the art (see, e.g., Bast et al. (1981) J Clin Invest. 68(5):1331-7; Scholler and Urban (2007) Biomark Med. 1(4):513-23; and Boudousq et al. (2013) PLoS One 8(7):e69613).

[0152] The term "human epidermal growth factor receptor 2," "HER2," or "HER2 / NEU," as used herein, refers to any naturally occurring form of human HER2. The term encompasses full-length HER2 (e.g., UniProt Reference Sequence: P04626; SEQ ID NO: 31) as well as any form of human HER2 that may result from cellular processing. The term also encompasses functional variants or fragments of human HER2, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human HER2 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). HER2 can be isolated from humans or produced recombinantly or by synthetic methods.

[0153] The term "anti-HER2 antibody" or "antibody that binds to HER2" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to HER2, and includes monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to HER2. U.S. Pat. No. 5,821,337 provides exemplary HER2-binding sequences, including exemplary anti-HER2 antibody sequences, and is incorporated herein by reference in this regard. In some embodiments, the anti-HER2 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment. Trastuzumab (U.S. Pat. No. 5,821,337; Molina et al. (2001) Cancer Res. 61(12):4744-9) is an exemplary anti-human HER2 antibody.

[0154] The terms "syndecan-1," "SDC1," or "CD138," as used herein, refer to any naturally occurring form of human CD138. The term encompasses full-length CD138 (e.g., UniProt Reference Sequence: P18827; SEQ ID NO: 32) as well as any form of human CD138 that may result from cellular processing. The term also encompasses functional variants or fragments of human CD138, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human CD138 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). CD138 can be isolated from humans or produced recombinantly or by synthetic methods.

[0155] The term "anti-CD138 antibody" or "antibody that binds to CD138" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to CD138, and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to CD138. In some embodiments, the anti-CD138 antibody used in the ADCs disclosed herein is an internalizing antibody or internalizing antibody fragment. B-B4 (Tassone et al. (2004) Blood 104:3688-96) is an exemplary anti-human CD138 antibody.

[0156] The term "ephrin type-A receptor 2" or "EPHA2," as used herein, refers to any naturally occurring form of human EPHA2. This term encompasses full-length EPHA2 (e.g., UniProt Reference Sequence: P29317; SEQ ID NO: 33) as well as any form of human EPHA2 that may result from cellular processing. This 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 included unless the context dictates that the term is used to refer only to the wild-type protein). EPHA2 can be isolated from humans or produced recombinantly or by synthetic methods.

[0157] The term "anti-EPHA2 antibody" or "antibody that binds to EPHA2" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to EPHA2, and includes monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to EPHA2. WO 2007 / 030642 provides exemplary EPHA2-binding sequences, including exemplary anti-EPHA2 antibody sequences, and is incorporated herein by reference in this regard. In some embodiments, the anti-EPHA2 antibody used in the ADCs disclosed herein is an internalizing antibody or internalizing antibody fragment. 1C1 (WO 2007 / 030642; Jackson et al. (2008) Cancer Res. 68(22):9367-74) is an exemplary anti-human EPHA2 antibody.

[0158] The term "mesothelin" or "MSLN," as used herein, refers to any naturally occurring form of human MSLN. This term encompasses full-length MSLN (e.g., UniProt Reference Sequence: Q13421; SEQ ID NO: 43) as well as any form of human MSLN that may result from cellular processing. This term also encompasses functional variants or fragments of human MSLN, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human MSLN (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). MSLN can be isolated from humans or produced recombinantly or by synthetic methods.

[0159] The term "anti-MSLN antibody" or "antibody that binds to MSLN" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to MSLN, and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to MSLN. WO 2011 / 074621 provides exemplary MSLN-binding sequences, including exemplary anti-MSLN antibody sequences, and is incorporated herein by reference. In some embodiments, the anti-MSLN antibody used in the ADCs disclosed herein is an internalizing antibody or internalizing antibody fragment. IC11-25, IC14-30, IC7-4, IC17-35, and 2-9 are exemplary anti-human MSLN antibodies.

[0160] The term "glutamate carboxypeptidase 2" or "FOLH1," as used herein, refers to any naturally occurring form of human FOLH1. This term encompasses full-length FOLH1 (e.g., UniProt Reference Sequence: Q04609; SEQ ID NO: 44) as well as any form of human FOLH1 that may result from cellular processing. This term also encompasses functional variants or fragments of human FOLH1, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human FOLH1 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). FOLH1 can be isolated from humans or produced recombinantly or by synthetic methods.

[0161] The term "anti-FOLH1 antibody" or "antibody that binds to FOLH1" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to FOLH1, and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to FOLH1. WO 2019 / 012260 and WO 2017 / 212250 provide exemplary FOLH1-binding sequences, including exemplary anti-FOLH1 antibody sequences, and are incorporated herein by reference in their entirety. In some embodiments, the anti-FOLH1 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment. J591 (deimmunized) is an exemplary anti-human FOLH1 antibody.

[0162] The term "cadherin-6" or "CDH6," as used herein, refers to any naturally occurring form of human CDH6. The term encompasses full-length CDH6 (e.g., UniProt Reference Sequence: P55285; SEQ ID NO: 45) as well as any form of human CDH6 that may result from cellular processing. The term also encompasses functional variants or fragments of human CDH6, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human CDH6 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). CDH6 can be isolated from humans or produced recombinantly or by synthetic methods.

[0163] The terms "anti-CDH6 antibody" or "antibody that binds to CDH6" refer to any form of antibody or fragment thereof that binds, e.g., specifically, to CDH6, and encompass monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to CDH6. WO 2018 / 185618 provides exemplary CDH6-binding sequences, including exemplary anti-CDH6 antibody sequences, and is incorporated herein by reference in this regard. In some embodiments, the anti-CDH6 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0164] The term "carcinoembryonic antigen-related cell adhesion molecule 5" or "CEACAM5," as used herein, refers to any naturally occurring form of human CEACAM5. This term encompasses full-length CEACAM5 (e.g., UniProt Reference Sequence: P06731; SEQ ID NO: 46) as well as any form of human CEACAM5 that may result from cellular processing. This term also encompasses functional variants or fragments of human CEACAM5, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human CEACAM5 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). CEACAM5 can be isolated from humans or produced recombinantly or by synthetic methods.

[0165] The terms "anti-CEACAM5 antibody" or "antibody that binds to CEACAM5" refer to any form of antibody or fragment thereof that binds, e.g., specifically, to CEACAM5, and include monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to CEACAM5. U.S. Patent Application Publication No. 2015 / 0125386 provides exemplary CEACAM5 binding sequences, including exemplary anti-CEACAM5 antibody sequences, and is incorporated herein by reference in this regard. In some embodiments, the anti-CEACAM5 antibody used in the ADCs disclosed herein is an internalizing antibody or internalizing antibody fragment. hMN14 is an exemplary anti-human CEACAM5 antibody.

[0166] The term "cryptic family protein 1B" or "CFClB" as used herein refers to any naturally occurring form of human CFClB. This term encompasses full-length CFClB (e.g., UniProt Reference Sequence: P0CG36; SEQ ID NO: 47) as well as any form of human CFClB that may result from cellular processing. This term also encompasses functional variants or fragments of human CFClB, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human CFClB (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). CFClB can be isolated from humans or produced recombinantly or by synthetic methods.

[0167] The terms "anti-CFC1B antibody" or "antibody that binds to CFC1B" refer to any form of antibody or fragment thereof that binds, e.g., specifically, to CFC1B, and encompass monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to CFC1B. WO 2002 / 088170 provides exemplary CFC1B-binding sequences, including exemplary anti-CFC1B antibody sequences, and is incorporated herein by reference in this regard. In some embodiments, the anti-CFC1B antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0168] The term "ectonucleotide pyrophosphatase / phosphodiesterase family member 3" or "ENPP3," as used herein, refers to any naturally occurring form of human ENPP3. This term encompasses full-length ENPP3 (e.g., UniProt Reference Sequence: O14638; SEQ ID NO: 48) as well as any form of human ENPP3 that may result from cellular processing. This term also encompasses functional variants or fragments of human ENPP3, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human ENPP3 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). ENPP3 can be isolated from humans or produced recombinantly or by synthetic methods.

[0169] The terms "anti-ENPP3 antibody" or "antibody that binds to ENPP3" refer to any form of antibody or fragment thereof that binds, e.g., specifically, to ENPP3, and encompass monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to ENPP3. Donate et al. ((2016) Clin Cancer Res. 22(8):1989-99) provide exemplary ENPP3-binding sequences, including exemplary anti-ENPP3 antibody sequences, and are incorporated herein by reference in their entirety. In some embodiments, the anti-ENPP3 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0170] The term "folate receptor alpha" or "FOLR1," as used herein, refers to any naturally occurring form of human FOLR1. The term encompasses full-length FOLR1 (e.g., UniProt Reference Sequence: P15328; SEQ ID NO: 49) as well as any form of human FOLR1 that may result from cellular processing. The term also encompasses functional variants or fragments of human FOLR1, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human FOLR1 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). FOLR1 can be isolated from humans or produced recombinantly or by synthetic methods.

[0171] The term "anti-FOLR1 antibody" or "antibody that binds to FOLR1" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to FOLR1, and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to FOLR1. WO 2005 / 080431 and Coney et al. ((1991) Cancer Res. 51(22):6125-32) provide exemplary FOLR1-binding sequences, including exemplary anti-FOLR1 antibody sequences, and are incorporated herein by reference in their entirety. In some embodiments, the anti-FOLR1 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment. Farletuzumab and MOv19 are exemplary anti-human FOLR1 antibodies.

[0172] The term "hepatitis A virus cellular receptor 1" or "HAVCR1," as used herein, refers to any naturally occurring form of human HAVCR1. This term encompasses full-length HAVCR1 (e.g., UniProt Reference Sequence: Q96D42; SEQ ID NO: 50) as well as any form of human HAVCR1 that may result from cellular processing. This term also encompasses functional variants or fragments of human HAVCR1, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human HAVCR1 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). HAVCR1 can be isolated from humans or produced recombinantly or by synthetic methods.

[0173] The terms "anti-HAVCR1 antibody" or "antibody that binds to HAVCR1" refer to any form of antibody or fragment thereof that binds, e.g., specifically, to HAVCR1, and encompass monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to HAVCR1. Thomas et al. ((2016) Mol Cancer Ther. 15(12):2946-54) provides exemplary HAVCR1-binding sequences, including exemplary anti-HAVCR1 antibody sequences, and is incorporated herein by reference in its entirety. In some embodiments, the anti-HAVCR1 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0174] The term "mast / stem cell growth factor receptor Kit" or "KIT," as used herein, refers to any naturally occurring form of human KIT. The term encompasses full-length KIT (e.g., UniProt Reference Sequence: P10721; SEQ ID NO: 51) as well as any form of human KIT that may arise by cellular processing. The term also encompasses functional variants or fragments of human KIT, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human KIT (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). KIT can be isolated from humans or produced recombinantly or by synthetic methods.

[0175] The term "anti-KIT antibody" or "antibody that binds to KIT" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to KIT, and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to KIT. Shi et al. ((2016) Proc Natl Acad Sci USA 113(33):E4784-93) and Abrams et al. ((2018) Clin Cancer Res. 24(17):4297-308) provide exemplary KIT-binding sequences, including exemplary anti-KIT antibody sequences, and are incorporated herein by reference in their entirety. In some embodiments, the anti-KIT antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0176] The term "hepatocyte growth factor receptor" or "MET," as used herein, refers to any naturally occurring form of human MET. The term encompasses full-length MET (e.g., UniProt Reference Sequence: P08581; SEQ ID NO: 52) as well as any form of human MET that may result from cellular processing. The term also encompasses functional variants or fragments of human MET, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human MET (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). MET can be isolated from humans or produced recombinantly or by synthetic methods.

[0177] The term "anti-MET antibody" or "antibody that binds to MET" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to MET, and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to MET. Yang et al. ((2019) Acta Pharmacol Sin.) provides exemplary MET-binding sequences, including exemplary anti-MET antibody sequences, and is incorporated herein by reference in this regard. In some embodiments, the anti-MET antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0178] The term "mucin-16" or "MUC16," as used herein, refers to any naturally occurring form of human MUC16. This term encompasses full-length MUC16 (e.g., UniProt Reference Sequence: Q8WXI7; SEQ ID NO:53) as well as any form of human MUC16 that may result from cellular processing. This term also encompasses functional variants or fragments of human MUC16, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human MUC16 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). MUC16 can be isolated from humans or produced recombinantly or by synthetic methods.

[0179] The terms "anti-MUC16 antibody" or "antibody that binds to MUC16" refer to any form of antibody or fragment thereof that binds, e.g., specifically, to MUC16, and encompass monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to MUC16. Liu et al. ((2016) Ann Oncol. 27(11):2124-30) provide exemplary MUC16-binding sequences, including exemplary anti-MUC16 antibody sequences, and are incorporated herein by reference in their entirety. In some embodiments, the anti-MUC16 antibody used in the ADCs disclosed herein is an internalizing antibody or internalizing antibody fragment.

[0180] The term "zinc transporter ZIP6" or "SLC39A6," as used herein, refers to any naturally occurring form of human SLC39A6. This term encompasses full-length SLC39A6 (e.g., UniProt Reference Sequence: Q13433; SEQ ID NO: 54) as well as any form of human SLC39A6 that may result from cellular processing. This term also encompasses functional variants or fragments of human SLC39A6, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human SLC39A6 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). SLC39A6 can be isolated from humans or produced recombinantly or by synthetic methods.

[0181] The term "anti-SLC39A6 antibody" or "antibody that binds to SLC39A6" refers to any form of antibody or fragment thereof that binds, e.g., specifically, to SLC39A6, and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to SLC39A6. Sussman et al. ((2014) Mol Cancer Ther. 13(12):2991-3000) provides exemplary SLC39A6-binding sequences, including exemplary anti-SLC39A6 antibody sequences, and is incorporated herein by reference in its entirety. In some embodiments, the anti-SLC39A6 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0182] The term "choline transporter-like protein 4" or "SLC44A4," as used herein, refers to any naturally occurring form of human SLC44A4. This term encompasses full-length SLC44A4 (e.g., UniProt Reference Sequence: Q53GD3; SEQ ID NO: 55) as well as any form of human SLC44A4 that may result from cellular processing. This term also encompasses functional variants or fragments of human SLC44A4, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human SLC44A4 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). SLC44A4 can be isolated from humans or produced recombinantly or by synthetic methods.

[0183] The terms "anti-SLC44A4 antibody" or "antibody that binds to SLC44A4" refer to any form of antibody or fragment thereof that binds, e.g., specifically, to SLC44A4, and encompass monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to SLC44A4. Mattie et al. ((2016) Mol Cancer Ther. 15(11):2679-87) provide exemplary SLC44A4 binding sequences, including exemplary anti-SLC44A4 antibody sequences, and are incorporated herein by reference in their entirety. In some embodiments, the anti-SLC44A4 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0184] The term "metalloreductase STEAP1" or "STEAP1," as used herein, refers to any naturally occurring form of human STEAP1. This term encompasses full-length STEAP1 (e.g., UniProt Reference Sequence: Q9UHE8; SEQ ID NO: 56) as well as any form of human STEAP1 that may result from cellular processing. This term also encompasses functional variants or fragments of human STEAP1, including, but not limited to, splice variants, allelic variants, and isoforms that retain one or more biological functions of human STEAP1 (i.e., variants and fragments are included unless the context dictates that the term is used to refer only to the wild-type protein). STEAP1 can be isolated from humans or produced recombinantly or by synthetic methods.

[0185] The terms "anti-STEAP1 antibody" or "antibody that binds to STEAP1" refer to any form of antibody or fragment thereof that binds, e.g., specifically, to STEAP1, and encompass monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and antibody fragments that are biologically functional so long as they bind, e.g., specifically, to STEAP1. WO 2008 / 052187 provides exemplary STEAP1-binding sequences, including exemplary anti-STEAP1 antibody sequences, and is incorporated herein by reference in this regard. In some embodiments, the anti-STEAP1 antibody used in the ADCs disclosed herein is an internalizing antibody or an internalizing antibody fragment.

[0186] As used herein, the terms "specific," "specifically binds," and "binds specifically" refer to the binding reaction between an antibody or antigen-binding fragment (e.g., an anti-HER2 antibody) and a target antigen (e.g., HER2) in a heterogeneous population of proteins and other biological substances. An antibody can be tested for binding specificity by comparing binding to the appropriate antigen with binding to an irrelevant antigen or antigen mixture under a given set of conditions. An antibody is considered specific if it binds to the appropriate antigen with at least 2-, 5-, 7-, and preferably 10-fold or more higher affinity than the irrelevant antigen or antigen mixture. A "specific antibody" or "target-specific antibody" is one that binds only to the target antigen (e.g., HER2) and does not bind to (or exhibits only minimal binding to) other antigens. In certain embodiments, an antibody or antigen-binding fragment that specifically binds to a target antigen (e.g., HER2) has a binding affinity of 1×10 -6 Less than M, 1 x 10 -7 Less than M, 1 x 10 -8 Less than M, 1 x 10 -9 Less than M, 1 x 10 -10 Less than M, 1 x 10 -11 Less than M, 1 x 10 -12 Less than M or 1 x 10 -13 K less than M D In certain embodiments, KD In some embodiments, K D is 500 pM to 1 μM, 1 μM to 100 nM, or 100 mM to 10 nM.

[0187] The term "epitope" refers to a portion of an antigen that has the ability to be recognized and specifically bound by an antibody. When the antigen is a polypeptide, the epitope can be formed by contiguous amino acids or by non-contiguous amino acids that are adjacent to each other when the polypeptide folds into a tertiary structure. The epitope bound by an antibody can be identified using any epitope mapping technique known in the art, including X-ray crystallography for epitope identification by direct visualization of the antigen-antibody complex, and monitoring the binding of antibody to fragments or mutant variants of the antigen, or monitoring the solvent accessibility of various portions of the antibody and antigen. Exemplary strategies used to map 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) 21:145-56; and Hager-Braun and Tomer (2005) Expert Rev Proteomics 2:745-56).

[0188] Competitive binding and epitope binning can also be used to determine antibodies that share the same or overlapping epitopes. Competitive binding is described in "Antibodies, A Laboratory Manual," Cold Spring Harbor Laboratory, Harlow and Lane (1999). st Edition 1988,2 ndCompetitive binding can be assessed using a cross-blocking assay, such as that described in the Journal of Molecular Biology, Vol. 1, pp. 2014 (2014). In some embodiments, competitive binding is identified when binding to a target antigen, such as HER2, by a reference antibody or binding protein (e.g., a binding protein comprising a CDR and / or variable domain selected from those identified in Tables 2-4) is reduced by the test antibody or binding protein by at least about 50% (e.g., 50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.5%, or more, or any percentage in between) in a cross-blocking assay. In some embodiments, competitive binding can be due to shared or similar (e.g., partially overlapping) epitopes, or due to steric hindrance of antibodies or binding proteins binding to nearby epitopes (see, e.g., Tzartos, Methods in Molecular Biology (Morris, ed. (1998) vol. 66, pp. 55-66)). In some embodiments, competitive binding can be used to sort 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 separated into another group of binding proteins that do not have overlapping or nearby epitopes.

[0189] The term "k" on " or "k a " refers to the on-rate constant for the association of an antibody to an antigen to form an antibody / antigen complex. This rate can be determined using standard assays such as surface plasmon resonance, biolayer interferometry, or ELISA assays.

[0190] The term "k" off " or "k d " refers to the off-rate constant for dissociation of an antibody from the antibody / antigen complex. This rate can be determined using a standard assay such as surface plasmon resonance, biolayer interferometry, or an ELISA assay.

[0191] The term “K D " refers to the equilibrium dissociation constant of a particular antibody-antigen interaction. D is k a / k d This rate can be determined using standard assays such as surface plasmon resonance, biolayer interferometry, or ELISA assays.

[0192] 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 each antibody or antigen-binding fragment (Ab) in an ADC of Formula (I). In an ADC that includes a splicing modulator drug moiety, "p" refers to the number of splicing modulator compounds linked to the antibody or antigen-binding fragment. For example, if two splicing modulator compounds (e.g., two compounds each having the structure D1) are linked to the antibody or antigen-binding fragment, then p=2. In a composition comprising multiple copies of an ADC of Formula (I), "average p" refers to the average number of -LD moieties per each antibody or antigen-binding fragment, also referred to as "average drug loading."

[0193] "Linker" or "linker moiety" is used herein to refer to any chemical moiety capable of covalently tethering a compound, typically a drug moiety such as a splicing modulator drug moiety, to another moiety, such as an antibody or antigen-binding fragment. The linker can be susceptible to or substantially resistant to acid-induced cleavage, peptidase-induced cleavage, light-based cleavage, esterase-induced cleavage, and / or disulfide bond cleavage, under conditions where the compound or antibody remains active.

[0194] The term "agent" is used herein to refer to a chemical compound, a mixture of chemical compounds, a biological macromolecule, or an extract made from biological materials. The terms "therapeutic agent" or "drug" refer to an agent that has the ability to modulate a biological process and / or has biological activity. The splicing modulator compounds described herein are exemplary therapeutic agents.

[0195] The term "chemotherapeutic agent" or "anti-cancer agent" is used herein to refer to any agent effective in treating cancer, regardless of mechanism of action. Inhibition of metastasis or angiogenesis is frequently a property of chemotherapeutic agents. Chemotherapeutic agents include antibodies, biomolecules, and small molecules, including the splicing modulator compounds described herein. Chemotherapeutic agents can be cytotoxic or cytostatic. The term "cytostatic agent" refers to an agent that inhibits or suppresses cell growth and / or proliferation. The term "cytotoxic agent" refers to a substance that causes cell death primarily by interfering with cellular expression activity and / or function.

[0196] As used herein, the terms "splice modulating agent," "spliceosome modulating agent," or "splice modulating agent" refer to compounds that have anti-cancer activity by interacting with components of the spliceosome. In some embodiments, splicing modulating agents alter the rate or type of splicing in target cells. For example, splicing modulating agents that function as inhibitory agents have the ability to reduce uncontrolled cell proliferation. In some embodiments, splicing modulating agents may act by binding to the SF3b spliceosome complex. Such modulating agents may be natural or synthetic compounds. Non-limiting examples of splicing modulating agents and classes of such modulating agents include pladienolides (e.g., pladienolide D or pladienolide B), pladienolide derivatives (e.g., pladienolide D or pladienolide B derivatives), herboxidiene, herboxidiene derivatives, splicostatin, splicostatin derivatives, sudemycin, or sudemycin derivatives. As used herein, the terms "derivative" and "analog," when referring to splicing modulators and the like, refer to any such compound that retains essentially the same, similar, or enhanced biological function or activity as the original compound, but with an altered chemical or biological structure. In some embodiments, the splicing modulator is a pladienolide or a pladienolide derivative.

[0197] As used herein, "pladienolide derivatives" refer to compounds structurally related to members of the family of natural products known as pladienolides, but retain one or more biological functions of the starting compound. Pladienolides were originally identified in the bacterium Streptomyces platensis (Mizui et al. (2004) J Antibiot. 57:188-96) as potentially cytotoxic compounds that cause cell cycle arrest in the G1 and G2 / M phases of the cell cycle (e.g., Bonnal et al. (2012) Nat Rev Drug Dis 11:847-59). There are seven naturally occurring pladienolides, pladienolides A through G (Mizui et al. (2004) J Antibiot. 57:188-96; Sakai et al. (2004) J Antibiotics 57:180-7). U.S. Patent Nos. 7,884,128 and 7,816,401 describe exemplary synthetic methods for pladienolides B and D, and are each incorporated herein by reference for such methods. Pladienolides B and D can also be synthesized using the exemplary method described in Kanada et al. ((2007) Angew Chem Int Ed. 46:4350-5). Kanada et al. and WO 2003 / 099813 describe an exemplary synthetic method for E7107 (D11) (compound 45 in WO 2003 / 099813) from pladienolide D (11107D in WO 2003 / 099813). The corresponding U.S. Patent Number is U.S. Patent No. 7,550,503 to Kotake et al. Each of these references is incorporated herein by reference for the synthetic methods described.

[0198] As used herein, "splice modulator drug moiety" refers to the component of an ADC or composition that provides the structure of the splicing modulator compound, e.g., the splicing modulator (D) component in an ADC of Formula (I) or in a composition comprising -LD.

[0199] As used herein, "spliceosome" refers to a ribonucleoprotein complex that removes introns from one or more RNA segments, such as pre-mRNA segments.

[0200] The term "homologue" refers to a molecule that exhibits homology to another molecule, for example, by having sequences of chemical residues that are the same or similar at corresponding positions.

[0201] The terms "inhibit" or "inhibition of," as used herein, mean to reduce by a measurable amount and can include, but need not necessarily, complete prevention or inhibition.

[0202] The terms "target negative," "target antigen negative," or "antigen negative" refer to the absence of target antigen expression by a cell or tissue. The terms "target positive," "target antigen positive," or "antigen positive" refer to the presence of target antigen expression. For example, a cell or cell line that does not express a target antigen may be described as target negative, while a cell or cell line that expresses the target antigen may be described as target positive.

[0203] The term "bystander killing" or "bystander effect" refers to the killing of target-negative cells in the presence of target-positive cells, where killing of target-negative cells is not observed in the absence of target-positive cells. Cell-to-cell contact, or at least proximity, between target-positive and target-negative cells allows for bystander killing. This type of killing is distinguishable from "off-target killing," which refers to the indiscriminate killing of target-negative cells. "Off-target killing" can be observed even in the absence of target-positive cells.

[0204] The terms "neoplastic disorder" and "cancer" are used interchangeably herein to refer to the presence of cells with characteristics typical of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and / or certain morphological characteristics. While cancer cells often take the form of a tumor or mass, such cells may exist alone within a subject or circulate in the bloodstream as independent cells, such as leukemia or lymphoma cells. The terms "neoplastic disorder" and "cancer" include all types of cancer and cancer metastases, including hematological malignancies, solid tumors, sarcomas, carcinomas, and other solid and non-solid tumor cancers. Hematological malignancies can include B-cell malignancies, cancers of the blood (leukemia), cancers of the plasma cells (myeloma, e.g., multiple myeloma), or cancers of the lymph nodes (lymphoma). Exemplary B-cell malignancies include chronic lymphocytic leukemia (CLL), follicular lymphoma, mantle cell lymphoma, and diffuse large B-cell lymphoma. Leukemias may include acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelocytic leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), and the like. Lymphomas may include Hodgkin's lymphoma and non-Hodgkin's lymphoma. Other hematological malignancies may include myelodysplastic syndromes (MDS). Solid tumors may include carcinomas such as adenocarcinomas, e.g., breast cancer, pancreatic cancer, prostate cancer, colon or colorectal cancer, lung cancer, gastric cancer, cervical cancer, endometrial cancer, ovarian cancer, cholangiocarcinoma, glioma, melanoma, and the like.

[0205] The terms "tumor" and "neoplasm" refer to any mass of tissue resulting from excessive cell growth or proliferation, either benign or malignant, including precancerous lesions.

[0206] The terms "tumor cell" and "neoplastic cell" are used interchangeably and refer to individual cells or the entire population of cells derived from a tumor or neoplasm, including both non-tumorigenic cells and cancer stem cells. As used herein, the term "tumor cell" is modified by the term "non-tumorigenic" when referring simply to tumor cells that lack the capacity for regeneration and differentiation to distinguish them from cancer stem cells.

[0207] The terms "subject" and "patient" are used interchangeably herein to refer to any animal, such as any mammal, including but not limited to humans, non-human primates, rodents, etc. In some embodiments, the mammal is a mouse. In some embodiments, the mammal is a human. In some embodiments, the subject is a mouse. In some embodiments, the subject is a human.

[0208] The term "co-administration" or administration "in combination with" one or more therapeutic agents includes simultaneous and consecutive administration in any order.

[0209] A "pharmaceutical composition" refers to a preparation in a form that allows for administration and subsequently provides the intended biological activity of one or more active ingredients and / or achieves a therapeutic effect, and that does not contain additional components that are unacceptably toxic to the subject to which the formulation is administered. The pharmaceutical composition may be sterile.

[0210] "Pharmaceutical excipients" include materials such as adjuvants, carriers, pH adjusting and buffering agents, tonicity adjusting agents, wetting agents, preservatives, and the like.

[0211] "Pharmaceutically acceptable" means approved or expected to be approved by a federal or state regulatory agency for use in animals, and more particularly in humans, or listed in the United States Pharmacopoeia or other generally recognized pharmacopeia.

[0212] A "pharmaceutically acceptable salt" is a salt that retains the desired biological activity of the parent compound and does not impart undesired toxicological effects. Examples of such salts are (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 from elemental anions such as chlorine, bromine, and iodine. See, e.g., 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.

[0213] The term "effective amount," as used herein, refers to an amount of a compound, ADC, or composition (e.g., a splicing modulator or ADC) described herein that is sufficient to achieve a specifically described purpose, e.g., to produce a therapeutic effect after administration, such as a reduction in tumor growth rate or tumor volume, a decrease in cancer symptoms, or some other indication of therapeutic efficacy. An effective amount can be determined by routine methods related to the described purpose. The term "therapeutically effective amount" refers to an amount of a compound, ADC, or composition described herein that is effective to detectably kill, reduce, and / or inhibit tumor cell growth or spread, tumor size or number, and / or other measures of the level, stage, progression, and / or severity of cancer. A therapeutically effective amount may vary depending on the intended application (in vitro or in vivo), or the subject and disease state under treatment, e.g., the subject's weight and age, the severity of the disease state, the method of administration, etc., and can be readily determined by one of ordinary skill in the art. The term also applies to a dose that will induce a specific response in target cells, e.g., inhibition of cell growth. The specific dose may vary depending, for example, on the particular pharmaceutical composition, the subject and their age and pre-existing health condition or risk for a health condition, the dosing regimen to be followed, the severity of the disease, whether it is administered in combination with other agents, the timing of administration, the tissue to which it is administered, and the physical delivery system it is delivered in. In the case of cancer, a therapeutically effective amount of an ADC can reduce the number of cancer cells, reduce tumor size, inhibit (e.g., slow or stop) tumor metastasis, inhibit (e.g., slow or stop) tumor growth, and / or alleviate one or more symptoms.

[0214] A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used prior to or at an earlier stage of disease in a subject, the prophylactically effective amount will be less than the therapeutically effective amount.

[0215] As used herein, "treating" or "therapeutic" and grammatically related terms refer to any improvement in any outcome of a disease, such as prolonged survival, reduced morbidity, and / or reduced side effects resulting from alternative treatment modalities. As is readily understood in the art, complete eradication of the disease is encompassed, but is not required for therapeutic action. "Treatment" or "treating," as used herein, refers to the administration of a described ADC or composition to a subject, e.g., a patient. Treatment can be curing, healing, alleviating, mitigating, altering, ameliorating, palliating, improving, or affecting a disorder, such as cancer, a symptom of a disorder, or a predisposition to a disorder. In some embodiments, in addition to treating a subject with a condition, the compositions disclosed herein can also be provided prophylactically to prevent or reduce the likelihood of the condition developing.

[0216] In some embodiments, a labeled ADC is used. Suitable "labels" include radionuclides, enzymes, substrates, cofactors, inhibitors, fluorescent moieties, chemiluminescent moieties, magnetic particles, and the like.

[0217] As used herein, "protein" refers to at least two covalently linked amino acids. This term encompasses polypeptides, oligopeptides, and peptides. In some embodiments, the two or more covalently linked amino acids are linked by a peptide bond. For example, when a protein is recombinantly produced using an expression system and a host cell, the protein may be composed of naturally occurring amino acids and peptide bonds. Alternatively, the protein may contain synthetic amino acids (e.g., homophenylalanine, citrulline, ornithine, and norleucine) or "peptide or protein analogs" such as peptidomimetic structures, i.e., peptoids. Peptoids are an exemplary class of peptidomimetics whose side chains are attached to nitrogen atoms of the peptide backbone rather than the alpha carbon (as in amino acids), and have different hydrogen bonding and conformational characteristics compared to peptides (see, e.g., Simon et al. (1992) Proc Natl Acad Sci. USA 89:9367). As such, peptoids may be resistant to proteolysis or other physiological or storage conditions and may be effective at permeating cell membranes. Such synthetic amino acids may be incorporated by conventional methods well known in the art, particularly during in vitro antibody synthesis. In addition, any combination of peptidomimetics, synthetic, and naturally occurring residues / structures may be used. "Amino acid" also includes imino acid residues, such as proline and hydroxyproline. The amino acid "R group" or "side chain" may be in either the (L) or (S) configuration. In specific embodiments, the amino acid is in the (L) or (S) configuration.

[0218] A "recombinant protein" is a protein made using recombinant techniques, i.e., through the expression of a recombinant nucleic acid, using any techniques and methods known in the art. Methods and techniques for making recombinant proteins are well known in the art.

[0219] An "isolated" protein is one that is free from at least some of the materials that normally accompany it in its natural state, e.g., comprising at least about 5% by weight, or at least about 50% by weight, of the total protein in a given sample. It is understood that, in some circumstances, an isolated protein may comprise between 5% and 99.9% by weight of the total protein content. For example, the protein may be produced at significantly higher concentrations using an inducible promoter or a high-expression promoter, thus producing the protein at increased concentration levels. This definition includes production of antibodies in a wide variety of organisms and / or host cells known in the art.

[0220] For amino acid sequences, sequence identity and / or similarity may be determined using standard techniques known in the art, including, but not limited to, the local sequence identity algorithm of Smith and Waterman (1981) Adv Appl Math. 2:482, the sequence identity alignment algorithm of Needleman and Wunsch (1970) J Mol Biol. 48:443, the search for similarity method of Pearson and Lipman (1988) Proc Nat Acad Sci. USA 85:2444, computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Drive, Madison, Wis.), the BestFit sequence program described by Devereux et al. (1984) Nucl Acid Res. 12:387-95, preferably using default settings, or by visual inspection. Preferably, percent identity is calculated by FastDB based on the following parameters: mismatch penalty of 1; gap penalty of 1; gap size penalty of 0.33; and joining penalty of 30 ("Current Methods in Sequence Comparison and Analysis," Macromolecule Sequencing and Synthesis, Selected Methods and Applications, pp. 127-149 (1988), Alan R. Liss, Inc.).

[0221] An example of a useful algorithm is PILEUP. PILEUP creates a multiple sequence alignment from a group of related sequences using progressive pairwise alignments. It can also plot a tree showing the clustering relationships used to create the alignment. PILEUP uses a simplification of the progressive alignment method of Feng & Doolittle (1987) J Mol Evol. 35:351-60; this method is similar to that described by Higgins and Sharp (1989) CABIOS 5:151-3. Useful PILEUP parameters include a default gap weight of 3.00, a default gap length weight of 0.10, and weighted end gaps.

[0222] Another example of a useful algorithm is the BLAST algorithm described in Altschul et al. (1990) J Mol Biol. 215:403-10; Altschul et al. (1997) Nucl Acid Res. 25:3389-402; and Karin et al. (1993) Proc Natl Acad Sci. USA 90:5873-87. A particularly useful BLAST program is the WU-BLAST-2 program, which was derived from Altschul et al. (1996) Methods in Enzymology 266:460-80. WU-BLAST-2 uses several search parameters, most of which are set to default values. Adjustable parameters are set using the following values: overlap width = 1, overlap ratio = 0.125, and word threshold (T) = 11. The HSP S and HSP S2 parameters are dynamic values ​​that are established by the program itself depending on the composition of the particular sequence and the composition of the particular database in which the sequence of interest is being searched, however, these values ​​can be adjusted to increase sensitivity.

[0223] Another useful algorithm is Gapped BLAST as reported by Altschul et al. (1997) Nucl Acid Res. 25:3389-402. Gapped BLAST uses a BLOSUM-62 substitution score; a threshold T parameter set to 9; ungapped extension initiation by the 2-hit method, imposing a cost of 10+k for gap lengths of k; Xu set to 16, and Xg set to 40 for the database search step and 67 for the output step of the algorithm. Gapped alignments are initiated by scores corresponding to approximately 22 bits.

[0224] Generally, the amino acid homology, similarity, or identity between the proteins disclosed herein and variants thereof, for example, variants of target antigens (such as HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, or STEAP1) and variants of antibody variable domains (including individual variant CDRs) is at least 80% to the sequences set forth herein, for example, at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, nearly 100%, or 100% homology or identity.

[0225] Similarly, "percent (%) nucleic acid sequence identity" with respect to nucleic acid sequences of antibodies and other proteins identified herein is defined as the percentage of nucleotide residues in a candidate sequence that are identical with nucleotide residues in the coding sequence of the antigen-binding protein. An exemplary method utilizes the BLASTN module of WU-BLAST-2 set to default parameters, with overlap width and overlap rate set to 1 and 0.125, respectively.

[0226] Although the site or region where the amino acid sequence variation is introduced is predetermined, the mutation itself does not have to be predetermined. For example, to optimize the performance of a mutation at a given site, random mutagenesis can be performed at the target codon or region, and the expressed antigen-binding protein CDR mutants are screened for the optimal combination of desired activity. Techniques for creating substitution mutations at predetermined sites in DNA with a known sequence are well known, such as MI3 primer mutagenesis and PCR mutagenesis.

[0227] "Alkyl" or "alkyl group," as used herein, means a straight, branched, or cyclic hydrocarbon chain that is completely saturated. In certain embodiments, an alkyl group can contain 1 to 8 carbon atoms ("C1-C8 alkyl"). In certain embodiments, an alkyl group can contain 1 to 6 carbon atoms ("C1-C6 alkyl"). In certain embodiments, an alkyl group contains 1 to 3 carbon atoms. In still other embodiments, an alkyl group contains 2 to 3 carbon atoms, and in yet other embodiments, an alkyl group contains 1 to 2 carbon atoms.

[0228] "Alkylalkoxy," as used herein, refers to an alkyl group substituted with an alkoxy group. "Alkoxy," as used herein, refers to an alkyl group, as defined above, attached to the carbon backbone by an oxygen ("alkoxy") atom.

[0229] "Alkylamino," as used herein, refers to an alkyl group substituted with an amino group. "Amino," as used herein, refers to -NH, -NH(alkyl), or -N(alkyl)(alkyl).

[0230] "Alkylhydroxy" as used herein refers to an alkyl group substituted with an amino group. "Hydroxy" or "hydroxyl" as used herein refers to -OH.

[0231] "Alkylene" refers to a divalent radical of an alkyl group. For example, -CH-, -CHCH-, -CHCHCH-, -CHCHCHCH-, -CHCHCHCHCHCH-, and -CHCHCHCHCHCHCHCH- refer to methylene, ethylene, n-propylene, n-butylene, n-pentylene, and n-hexylene, respectively.

[0232] "Carbocycle," as used herein, includes both aromatic (e.g., aryl) and non-aromatic (e.g., cycloalkyl) groups. In certain embodiments, a carbocyclic group contains 3 to 10 carbon atoms (a "3- to 10-membered carbocyclic ring"). In certain embodiments, a carbocyclic group contains 3 to 8 carbon atoms (a "3- to 8-membered carbocyclic ring"). In certain embodiments, a carbocyclic group contains 3 to 6 carbon atoms (a "3- to 6-membered carbocyclic ring"). In certain embodiments, a carbocyclic group contains 3 to 5 carbon atoms (a "3- to 5-membered carbocyclic ring").

[0233] "Halogen" refers to any halogen radical, for example, -F, -Cl, -Br, or -I.

[0234] The terms "heterocycle," "heterocyclyl," and "heterocyclic," as used herein, mean a monocyclic heterocycle, a bicyclic heterocycle, or a tricyclic heterocycle containing at least one heteroatom in the ring.

[0235] Monocyclic heterocycles are 3-, 4-, 5-, 6-, 7-, or 8-membered rings containing at least one heteroatom independently selected from O, N, and S. In some embodiments, heterocycles are 3- or 4-membered rings containing one heteroatom selected from O, N, and S. In some embodiments, heterocycles are 5-membered rings containing zero or one double bond and one, two, or three heteroatoms selected from O, N, and S. In some embodiments, heterocycles are 6-, 7-, or 8-membered rings containing zero, one, or two double bonds and one, two, or three heteroatoms selected from O, N, and S. Representative examples of monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, dihydropyranyl (including 3,4-dihydro-2H-pyran-6-yl), 1,3-dithiolanyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl , oxazolinyl, oxazolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl (including tetrahydro-2H-pyran-4-yl), tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxidethiomorpholinyl (thiomorpholinesulfone), thiopyranyl, and trithianyl.

[0236] Bicyclic heterocycles of the present disclosure can include monocyclic heterocycles fused to an aryl group having a total of 5 to 12 ring atoms, or monocyclic heterocycles fused to a monocyclic cycloalkyl, or monocyclic heterocycles fused to a monocyclic cycloalkenyl, or monocyclic heterocycles fused to a monocyclic heterocycle. Examples of bicyclic heterocycles include, but are not limited to, 3,4-dihydro-2H-pyranyl, 1,3-benzodioxolyl, 1,3-benzodithiolyl, 2,3-dihydro-1,4-benzodioxinyl, 2,3-dihydro-1-benzofuranyl, 2,3-dihydro-1-benzothienyl, 2,3-dihydro-1H-indolyl, and 1,2,3,4-tetrahydroquinolinyl.

[0237] The terms "heterocycle," "heterocyclyl," and "heterocyclic" encompass heteroaryl. "Heteroaryl" refers to a cyclic moiety having one or more closed rings, at least one of which contains one or more heteroatoms (oxygen, nitrogen, or sulfur), wherein at least one of the rings is aromatic, and one or more of the rings can be independently fused and / or bridged. Examples include, without limitation, phenyl, thiophenyl, triazolyl, pyridinyl, pyrimidinyl, pyridazinyl, and pyrazinyl.

[0238] As described herein, compounds of the present disclosure may contain "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be either the same or different at each position. Combinations of substituents envisioned by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds.

[0239] Those of ordinary skill in the art will understand that "substituted" or "substituted with" or "absence" includes the implicit proviso that such substitution or absence is subject to the allowed valences of the substituted atom and substituents, and that the substitution or absence results in a stable compound, e.g., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, etc. For purposes of this disclosure, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valences of the heteroatom.

[0240] "Stable" refers to a compound that does not substantially change chemically and / or physically when subjected to conditions that allow for its production, detection, and in certain embodiments, its recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is one that does not substantially change when placed at a temperature of 40° C. or less in the absence of moisture or other chemically reactive conditions for at least one week. In some embodiments, the compounds disclosed herein are stable.

[0241] Enantiomers as taught herein may include "enantiopure" isomers that contain substantially a single enantiomer at a particular asymmetric center or centers, for example, greater than 90%, 92%, 95%, 98%, or 99%, or even 100%, of a single enantiomer. An "asymmetric center" or "chiral center" refers to a tetrahedral carbon atom containing four different substituents.

[0242] The compounds described herein may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may contain, for example, deuterium ( 2 H), tritium ( 3 H), carbon-13 ( 13 C), or carbon-14 ( 14C). All isotopic variations of the compounds disclosed herein, whether radioactive or not, are intended to be encompassed within the scope of the disclosure. Additionally, all tautomeric forms of the compounds described herein are intended to be within the scope of the claimed disclosure.

[0243] Antibody-drug conjugates Antibody-drug conjugate (ADC) compounds of the present disclosure include those with anti-cancer activity. Specifically, the ADC compounds comprise an antibody or antigen-binding fragment (including an antigen-binding fragment thereof) conjugated (i.e., covalently attached via a linker) to a drug moiety (e.g., a splicing modulator), wherein the drug moiety has a cytotoxic or cytostatic effect when not conjugated to the antibody or antigen-binding fragment. In various embodiments, the drug moiety has the ability to bind to and / or interact with the SF3b spliceosome complex when not conjugated to the antibody or antigen-binding fragment. In various embodiments, the drug moiety has the ability to modulate in vitro and / or in vivo RNA splicing when not conjugated to the antibody or antigen-binding fragment. By targeting RNA splicing, in various embodiments, the drug moieties and ADCs disclosed herein are potent anti-proliferative agents. In various embodiments, the drug moieties and ADCs disclosed herein can target both actively dividing and quiescent cells.

[0244] In various embodiments, the present disclosure is based, at least in part, on the discovery that certain biologically active splicing modulators can confer improved properties when used in ADCs. While a splicing modulator may exhibit desirable improved characteristics (e.g., robust SF3b spliceosome complex binding, potent RNA splicing modulation) when used alone, in various embodiments, a splicing modulator may exhibit a reduction in the same desirable improved characteristics when conjugated to an antibody or antigen-binding fragment. Thus, the development and production of ADCs for use as human therapeutics, e.g., oncology agents, may require more than identifying an antibody that binds to a desired target or targets and has the ability to link to a drug used alone to treat cancer. Linking an antibody to a drug can have a profound effect on the activity of either or both the antibody and the drug, which will vary depending on the type of linker and / or drug selected. 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-to-antibody ratio; (iv) enable delivery of the drug moiety by stable association with the antibody or antigen-binding fragment, e.g., intracellular delivery; (v) maintain stability of the ADC as an intact conjugate until transported or delivered to the target site; (vi) minimize aggregation of the ADC before or after administration; (vii) realize the therapeutic effect, e.g., cytotoxic effect, of the drug moiety after cleavage or other release mechanism in the cellular environment; (viii) exhibit in vivo anti-cancer therapeutic efficacy that is comparable to or superior to that of the antibody and drug moiety alone; (ix) minimize off-target killing by the drug moiety; and / or (x) exhibit desirable pharmacokinetic and pharmacodynamic properties, formulation amenability, and toxicological / immunological profile. Each of these properties may be necessary for the identification of improved ADCs for therapeutic use (Ab et al. (2015) Mol Cancer Ther. 14:1605-13).

[0245] In various embodiments, the ADCs disclosed herein unexpectedly exhibit advantageous properties in some or each of the above-listed categories. For example, in some embodiments, the ADC constructs disclosed herein unexpectedly exhibit advantageous drug loading, aggregation, and / or stability profiles, and / or maintain antibody binding function, drug activity, and / or improved bystander killing while reducing off-target killing, when compared to ADCs comprising alternative linkers and / or drug moieties (e.g., alternative splicing modulators). In some embodiments, the ADC constructs disclosed herein demonstrate superior stability, activity, potency, or other efficacy (measured in vivo or in vitro) when compared to ADCs using alternative linkers and / or drug moieties (e.g., alternative splicing modulators). In some embodiments, the ADC constructs disclosed herein exhibit in vivo therapeutic efficacy when administered as a single dose. In some embodiments, the ADC constructs disclosed herein are unexpectedly stable when compared to ADCs that use alternative linkers and / or drug moieties (e.g., alternative splicing modulators).

[0246] The disclosed ADC compounds can selectively deliver effective doses of cytotoxic or cytostatic agents to cancer cells or tumor tissues. The disclosed ADCs have been found to have potent cytotoxic and / or cytostatic activity against cells expressing their respective target antigens (e.g., HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, STEAP1). In some embodiments, the cytotoxic and / or cytostatic activity of an ADC depends on the target antigen expression of the cells. In some embodiments, the disclosed ADCs are particularly effective at killing cancer cells that express the target antigen while minimizing off-target killing. In some embodiments, the disclosed ADCs do not exhibit cytotoxic and / or cytostatic effects against cancer cells that do not express the target antigen.

[0247] Exemplary cancers that express HER2 include, but are not limited to, breast cancer, gastric cancer, bladder cancer, urothelial cell carcinoma, esophageal cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial carcinoma), salivary duct cancer, cervical cancer, endometrial cancer, and ovarian cancer (English et al. (2013) Mol Diagn Ther. 17:85-99).

[0248] Exemplary cancers that express CD138 include, but are not limited to, intrathoracic cancers (e.g., lung cancer, mesothelioma), skin cancers (e.g., basal cell carcinoma, squamous cell carcinoma), head and neck cancers (e.g., larynx, hypopharynx, nasopharynx), breast cancer, genitourinary cancers (e.g., cervical cancer, ovarian cancer, endometrial cancer, prostate cancer, bladder cancer, urothelial carcinoma), hematological malignancies (e.g., myeloma such as multiple myeloma, B-cell malignancies, Hodgkin's lymphoma), and thyroid cancer (Szatmari et al. (2015) Dis Markers 2015:796052).

[0249] Exemplary cancers that express EPHA2 include breast cancer, brain cancer, ovarian cancer, bladder cancer, pancreatic cancer, esophageal cancer, lung cancer, prostate cancer, melanoma, esophageal cancer, and gastric cancer (Tandon et al. (2011) Expert Opin Ther Targets 15(1):31-51).

[0250] In some embodiments, cleavage of the ADC releases the splicing modulator from the antibody or antigen-binding fragment and linker. In some embodiments, the linker and / or splicing modulator are designed to promote bystander killing (killing of neighboring cells). In some embodiments, the linker and / or splicing modulator are designed to promote bystander killing through cleavage after cellular internalization and diffusion of the linker-drug moiety and / or drug moiety alone to neighboring cells. In some embodiments, the linker enhances cellular internalization. In some embodiments, the linker is designed to minimize cleavage in the extracellular environment, thereby reducing toxicity to off-target tissues (e.g., non-cancerous tissues), while maintaining ADC binding to target tissues and bystander killing of cancerous tissues surrounding the target cancerous tissue that do not express the antigen targeted by the antibody or antigen-binding fragment of the ADC, but do express that antigen. In some embodiments, the drug moiety, or a catabolic product of the drug moiety produced by cleavage of the ADC, is designed to facilitate uptake by target cells or neighboring cells (i.e., cell permeability). Such drug moieties and catabolic products may be referred to herein as "bystander active," while drug moieties or catabolic products with reduced cell permeability may be referred to as "bystander inactive."

[0251] In some embodiments, the disclosed ADCs also demonstrate bystander killing activity but with reduced off-target cytotoxicity. Without being bound by theory, the bystander killing activity of an ADC may be particularly beneficial when its penetration into solid tumors is limited and / or when target antigen expression among tumor cells is heterogeneous. In some embodiments, ADCs comprising cleavable linkers are particularly effective at bystander killing and / or demonstrate improved bystander killing activity compared to comparable treatment with ADCs comprising non-cleavable linkers. In some embodiments, the ADCs disclosed herein exhibit improved solubility and target cell permeability compared to the drug moiety itself. In some embodiments, the ADCs disclosed herein exhibit improved cytotoxicity compared to the drug moiety itself. In some embodiments, the ADCs disclosed herein use drug moieties that exhibit low cytotoxicity when evaluated as solo drugs, but surprisingly outperform ADCs comprising other drug moieties that have high cytotoxicity when evaluated as solo drugs. In some embodiments, cleavage and release of the splicing modulator improves the cytotoxicity of the ADC compared to equivalent treatment with an ADC comprising a non-cleavable linker. In other embodiments, cleavage and release of the splicing modulator is not required for the ADC to have the desired biological activity. In some embodiments, ADCs comprising non-cleavable linkers with increased spacer length (e.g., ADL12) provide the same or comparable cytotoxicity compared to equivalent treatment with ADCs comprising cleavable linkers (e.g., ADL1, ADL5), and unexpectedly, provide superior cytotoxicity compared to equivalent treatment with ADCs comprising shorter non-cleavable linkers. In some embodiments, ADCs comprising non-cleavable linkers of increasing spacer length that do not contain a carbonyl group (e.g., ADL12) provide the same or comparable cytotoxicity compared to equivalent treatment with ADCs comprising cleavable linkers (e.g., ADL1, ADL5), and unexpectedly, provide superior cytotoxicity compared to equivalent treatment with ADCs comprising non-cleavable linkers with the same or comparable spacer length that contain a carbonyl group (e.g., ADL10).In some embodiments, removal of the carbonyl group from a non-cleavable MC linker (e.g., ADL12) can result in a greater than 50-fold, greater than 75-fold, greater than 100-fold, greater than 150-fold, or greater than 200-fold increase in cytotoxicity compared to equivalent treatment with an ADC comprising an unmodified non-cleavable MC linker (e.g., ADL10). In some embodiments, removal of the carbonyl group from a non-cleavable MC linker (e.g., ADL12) and increasing the spacer length (e.g., addition of at least one spacer unit) can result in a greater than 50-fold, greater than 75-fold, greater than 100-fold, greater than 150-fold, or greater than 200-fold increase in cytotoxicity compared to equivalent treatment with an ADC comprising an unmodified non-cleavable MC linker (e.g., ADL10).

[0252] Provided herein are ADC compounds comprising an antibody or antigen-binding fragment thereof (Ab) that targets tumor cells, a splicing modulator drug moiety (D), and a linker moiety (L) that covalently links the Ab to D. In certain aspects, the antibody or antigen-binding fragment is capable of binding to a tumor-associated antigen (e.g., HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, STEAP1) with high specificity and high affinity. In certain embodiments, the antibody or antigen-binding fragment is internalized within the target cell upon binding, e.g., into a degradable compartment within the cell. In various embodiments, ADCs may be used that internalize upon binding to target cells, undergo degradation, release the splicing modulator drug moiety, and kill cancer cells. The splicing modulator drug moiety may be released from the antibody and / or linker moiety of the ADC by enzymatic action, hydrolysis, oxidation, or any other mechanism.

[0253] An exemplary ADC has formula (I): Ab-(LD) p (I) where Ab = antibody or antigen-binding fragment, L = linker moiety, D = splicing modulator drug moiety, and p = number of splicing modulator drug moieties per each antibody or antigen-binding fragment. It has.

[0254] In certain preferred embodiments, the drug targeting moiety for use in the described ADCs and compositions is an antibody or antigen-binding fragment. Other exemplary drug targeting moieties for use in the described ADCs and compositions are also provided and described herein. In some embodiments, the drug targeting moiety can be any one of a variety of cell-binding agents and non-antibody scaffolds. In some embodiments, the drug targeting moiety is a cell-binding agent. As used herein, the term "cell-binding agent" refers to any agent capable of binding to animal (e.g., human) cells and delivering a drug moiety (e.g., a splicing modulator drug moiety as disclosed herein). This term encompasses the exemplary antibodies and antigen-binding fragments (e.g., monoclonal antibodies and fragments thereof, such as Fab and scFV) disclosed herein. The term further encompasses exemplary cell-binding agents, such as DARPins, duobodies, bicyclic peptides, nanobodies, centilins, MSH (melanocyte-stimulating hormone), receptor-Fc fusion molecules, T cell receptor structures, steroid hormones such as androgens and estrogens, growth factors, colony-stimulating factors such as EGF, and other non-antibody scaffolds. In various embodiments, non-antibody scaffolds can be broadly divided into two structural classes: domain-sized compounds (about 6-20 kDa) and constrained peptides (about 2-4 kDa). Exemplary domain-sized scaffolds include, but are not limited to, affibodies, affilins, anticalins, atrimers, DARPins, FN3 scaffolds (e.g., adnectins and sentinels), fynomers, Kunitz domains, pronectins, O-bodies, and receptor-Fc fusion proteins, while exemplary constrained peptides include avimers, bicyclic peptides, and Cys knots.In some embodiments, the drug targeting moiety used in the described ADCs and compositions is selected from an affibody, affilin, an anticalin, an atrimer, a DARPin, an FN3 scaffold such as an adnectin or sentinel, a fynomer, a Kunitz domain, a pronectin, an O-body, an avimer, a bicyclic peptide, and a Cys knot. In some embodiments, the drug targeting moiety used in the described ADCs and compositions is a receptor-Fc fusion protein, e.g., a HER2-Fc chimeric fusion protein. Non-antibody scaffolds are reviewed, for example, in Vazquez-Lombardi et al. (2015) Drug Dis Today 20(10):1271-83.

[0255] antibody The antibody or antigen-binding fragment (Ab) of formula (I) includes within its scope any antibody or antigen-binding fragment that specifically binds to a target antigen on a cancer cell. The antibody or antigen-binding fragment may have a dissociation constant (K) of ≦1 mM, ≦100 nM, or ≦10 nM, or any amount in between, as measured, for example, by BIAcore® analysis. D ) can bind to the target antigen. In certain embodiments, K D In some embodiments, K D is 500 pM to 1 μM, 1 μM to 100 nM, or 100 mM to 10 nM.

[0256] In some embodiments, the antibody or antigen-binding fragment is a four-chain antibody (also called an immunoglobulin or full-length antibody or intact antibody), comprising two heavy chains and two light chains. In some embodiments, the antibody or antigen-binding fragment is a two-chain half body (one light chain and one heavy chain) or antigen-binding fragment of an immunoglobulin. In some embodiments, the antibody or antigen-binding fragment is an antigen-binding fragment of an immunoglobulin that retains the ability to bind to a target cancer antigen and / or provide immunoglobulin function.

[0257] In some embodiments, the antibody or antigen-binding fragment is an antibody or antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment is an internalizing antibody or internalizing antigen-binding fragment thereof. In some embodiments, the internalizing antibody or internalizing antigen-binding fragment thereof binds to a target cancer antigen expressed on the surface of a cell and, upon binding, enters the cell. In some embodiments, the splicing modulator drug moiety of the ADC is released from the antibody or antigen-binding fragment of the ADC after the ADC has entered and is present in a cell expressing the target cancer antigen (i.e., after the ADC has been internalized), for example, by cleavage, degradation of the antibody or antigen-binding fragment, or any other suitable release mechanism.

[0258] The amino acid sequences of exemplary antibodies of the present disclosure are shown in Tables 2-4.

[0259] [Table 1]

[0260] [Table 2]

[0261] [Table 3]

[0262] [Table 4]

[0263] [Table 5]

[0264] [Table 6]

[0265] Table 7

[0266] Table 8

[0267] Table 9

[0268] Table 10

[0269] Table 11

[0270] Table 12

[0271] Table 13

[0272] Table 14

[0273] Table 15

[0274] Table 16

[0275] [Table 17]

[0276] [Table 18]

[0277] [Table 19]

[0278] [Table 20]

[0279] [Table 21]

[0280] [Table 22]

[0281] [Table 23]

[0282] In various embodiments, the ADCs disclosed herein can comprise any set of heavy and light chain variable domains listed in the table above, or can comprise a set of six CDR sequences from a set of heavy and light chains, for example, by grafting those six CDRs onto a human donor antibody framework of choice. In various embodiments, the ADCs disclosed herein can comprise a set of six CDR sequences from a set of heavy and light chains, for example, by grafting those six CDRs onto a human donor antibody framework of choice, such that the ADC can target its target cancer antigen (e.g., 1×10 -8 K less than M DThe ADCs may include amino acid sequences that are homologous to those listed in the table above, so long as they retain the ability to bind to the target polypeptide (e.g., at 200 bp) and retain one or more functional properties of the ADCs disclosed herein (e.g., the ability to internalize, modulate RNA splicing, inhibit cell growth, etc.).

[0283] In some embodiments, the ADC further comprises human heavy and light chain constant domains or fragments thereof. For example, the ADC may comprise a human IgG heavy chain constant domain (such as an IgG1) and a human kappa or lambda light chain constant domain. In various embodiments, the antibody or antigen-binding fragment of the described ADC comprises a human immunoglobulin G subtype 1 (IgG1) heavy chain constant domain together with a human Ig kappa light chain constant domain.

[0284] In various other embodiments, the target cancer antigen of the ADC is human epidermal growth factor receptor 2 (HER2).

[0285] In various embodiments, the anti-HER2 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 1, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 3; a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 4, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 5, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 6, as defined by the Kabat numbering system.

[0286] In various embodiments, the anti-HER2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the anti-HER2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 19 and the light chain variable region amino acid sequence of SEQ ID NO: 20, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-HER2 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 19 and / or a light chain variable region amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 20.

[0287] In various embodiments, the anti-HER2 antibody or antigen-binding fragment thereof is an internalizing antibody or internalizing antigen-binding fragment. In various embodiments, the anti-HER2 antibody comprises a human IgG1 heavy chain constant domain and a human Ig kappa light chain constant domain.

[0288] In various embodiments, the anti-HER2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 19, or a sequence at least 95% identical to SEQ ID NO: 19, and a light chain amino acid sequence of SEQ ID NO: 20, or a sequence at least 95% identical to SEQ ID NO: 20. In particular embodiments, the anti-HER2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 19 and a light chain amino acid sequence of SEQ ID NO: 20, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-HER2 antibody has a heavy chain amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 19, and a light chain amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 20. In various embodiments, the anti-HER2 antibody is trastuzumab, or an antigen-binding fragment thereof.

[0289] In various embodiments, the anti-HER2 antibody or antigen-binding fragment thereof comprises the three heavy chain CDRs and three light chain CDRs of trastuzumab, or wherein the CDRs comprise no more than 1, 2, 3, 4, 5, or 6 amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO: 1), HCDR2 (SEQ ID NO: 2), HCDR3 (SEQ ID NO: 3); LCDR1 (SEQ ID NO: 4), LCDR2 (SEQ ID NO: 5), and LCDR3 (SEQ ID NO: 6).

[0290] In various other embodiments, the target cancer antigen of the ADC is human syndecan-1 (CD138).

[0291] In various embodiments, the anti-CD138 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 7, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 8, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 9, a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 10, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 11, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 12, as defined by the Kabat numbering system.

[0292] In various embodiments, the anti-CD138 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 22. In some embodiments, the anti-CD138 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 21 and the light chain variable region amino acid sequence of SEQ ID NO: 22, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD138 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 21 and / or a light chain variable region amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 22.

[0293] In various embodiments, the anti-CD138 antibody or antigen-binding fragment thereof is an internalizing antibody or internalizing antigen-binding fragment. In various embodiments, the anti-CD138 antibody comprises a murine IgG2a heavy chain constant domain and a murine Ig kappa light chain constant domain. In various embodiments, the anti-CD138 antibody comprises a human IgG2a heavy chain constant domain and a human Ig kappa light chain constant domain.

[0294] In various embodiments, the anti-CD138 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 21, or a sequence at least 95% identical to SEQ ID NO: 21, and a light chain amino acid sequence of SEQ ID NO: 22, or a sequence at least 95% identical to SEQ ID NO: 22. In particular embodiments, the anti-CD138 antibody comprises a heavy chain amino acid sequence of SEQ ID NO: 21 and a light chain amino acid sequence of SEQ ID NO: 22, or a sequence at least 95% identical to the disclosed sequences. In some embodiments, the anti-CD138 antibody has a heavy chain amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 21 and a light chain amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 22. In various embodiments, the anti-CD138 antibody is B-B4, or an antigen-binding fragment thereof.

[0295] In various embodiments, the anti-CD138 antibody or antigen-binding fragment thereof comprises the three heavy chain CDRs and three light chain CDRs of B-B4, or wherein the CDRs comprise no more than 1, 2, 3, 4, 5, or 6 amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO: 7), HCDR2 (SEQ ID NO: 8), HCDR3 (SEQ ID NO: 9); LCDR1 (SEQ ID NO: 10), LCDR2 (SEQ ID NO: 11), and LCDR3 (SEQ ID NO: 12).

[0296] In various other embodiments, the target cancer antigen of the ADC is human ephrin type-A receptor 2 (EPHA2).

[0297] In various embodiments, the anti-EPHA2 antibody or antigen-binding fragment thereof comprises three heavy chain CDRs and three light chain CDRs as follows: a heavy chain CDR1 (HCDR1) consisting of SEQ ID NO: 13, a heavy chain CDR2 (HCDR2) consisting of SEQ ID NO: 14, a heavy chain CDR3 (HCDR3) consisting of SEQ ID NO: 15, a light chain CDR1 (LCDR1) consisting of SEQ ID NO: 16, a light chain CDR2 (LCDR2) consisting of SEQ ID NO: 17, and a light chain CDR3 (LCDR3) consisting of SEQ ID NO: 18, as defined by the Kabat numbering system.

[0298] In various embodiments, the anti-EPHA2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment thereof comprises the heavy chain variable region amino acid sequence of SEQ ID NO: 23 and the light chain variable region amino acid sequence of SEQ ID NO: 24, or a sequence at least 95% identical to a disclosed sequence. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment thereof has a heavy chain variable region amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 23 and / or a light chain variable region amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 24.

[0299] In various embodiments, the anti-EPHA2 antibody or antigen-binding fragment thereof is an internalizing antibody or internalizing antigen-binding fragment. In various embodiments, the anti-EPHA2 antibody comprises a human IgG1 heavy chain constant domain and a human Ig kappa light chain constant domain.

[0300] In various embodiments, the anti-EPHA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO:23, or a sequence at least 95% identical to SEQ ID NO:23, and a light chain amino acid sequence of SEQ ID NO:24, or a sequence at least 95% identical to SEQ ID NO:24. In particular embodiments, the anti-EPHA2 antibody comprises a heavy chain amino acid sequence of SEQ ID NO:23 and a light chain amino acid sequence of SEQ ID NO:24, 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 at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:23 and a light chain amino acid sequence at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO:24. In some embodiments, the anti-EPHA2 antibody comprises a heavy chain encoded by the nucleotide sequence of SEQ ID NO:23; and a light chain encoded by the nucleotide sequence of SEQ ID NO:24. In various embodiments, the anti-EPHA2 antibody is 1C1, or an antigen-binding fragment thereof.

[0301] In various embodiments, the anti-EPHA2 antibody or antigen-binding fragment thereof comprises the three heavy chain CDRs and three light chain CDRs of 1C1, or wherein the CDRs comprise no more than 1, 2, 3, 4, 5, or 6 amino acid additions, deletions, or substitutions of HCDR1 (SEQ ID NO: 13), HCDR2 (SEQ ID NO: 14), HCDR3 (SEQ ID NO: 15); LCDR1 (SEQ ID NO: 16), LCDR2 (SEQ ID NO: 17), and LCDR3 (SEQ ID NO: 18).

[0302] In various embodiments, amino acid substitutions are of single residues. Insertions can typically be on the order of about 1 to about 20 amino acid residues, although significantly larger insertions can be tolerated as long as biological function (e.g., binding to a target antigen) is maintained. Deletions typically range from about 1 to about 20 amino acid residues, although in some cases deletions can be much larger. Substitutions, deletions, insertions, or any combination thereof can be used to arrive at the final derivative or variant. Generally, these changes are made to a small number of amino acids to minimize changes to the molecule, particularly the immunogenicity and specificity of the antigen-binding protein. However, larger changes can be tolerated in certain circumstances. Generally, conservative substitutions are made according to the following chart, shown as Table 6:

[0303] [Table 24]

[0304] Substantial changes in function or immunological identity are made by selecting substitutions that are less conservative than those shown in Table 6. For example, substitutions can be made that more significantly affect the structure of the polypeptide backbone, e.g., α-helical or β-sheet structure, in the altered region; the charge or hydrophobicity of the molecule at the target site; or the bulk of the side chain. In general, the substitutions that are likely to produce the greatest changes in the properties of a polypeptide are those in which a hydrophilic residue, e.g., seryl or threonyl, is substituted for (or by) a hydrophobic residue, e.g., leucyl, isoleucyl, phenylalanyl, valyl, or alanyl; (b) cysteine ​​or proline is substituted for (or by) any other residue; (c) a residue with an electropositive side chain, e.g., lysyl, arginyl, or histidyl, is substituted for (or by) an electronegative residue, e.g., glutamyl or aspartyl; or (d) a residue with a bulky side chain, e.g., phenylalanine, is substituted for (or by) one without a side chain, e.g., glycine.

[0305] In various embodiments in which variant antibody sequences are used in ADCs, the variants will typically exhibit the same qualitative biological activity and elicit the same immune response, although variants may also be selected to modify the characteristics of the antigen-binding protein, if desired. Alternatively, variants may be designed to alter the biological activity of the antigen-binding protein. For example, glycosylation sites may be altered or removed.

[0306] Various antibodies can be used with the ADCs used herein to target cancer cells. As shown below, the linker-payloads of the ADCs disclosed herein are surprisingly effective with various tumor antigen-targeting antibodies. Suitable antigens expressed in tumor cells but not in healthy cells, or highly expressed in tumor cells compared to healthy cells, are known in the art, as are antibodies directed thereto. These antibodies can be used with the linkers and splicing modulator payloads disclosed herein. In some embodiments, the antibody or antigen-binding fragment targets HER2, and the HER2-targeting antibody or antigen-binding fragment is trastuzumab. In some embodiments, the antibody or antigen-binding fragment targets CD138, and the CD138-targeting antibody or antigen-binding fragment is B-B4. In some embodiments, the antibody or antigen-binding fragment targets EPHA2, and the EPHA2-targeting antibody or antigen-binding fragment is 1C1. In some embodiments, the disclosed linkers and splicing modulator payloads are surprisingly effective with several different tumor-targeting antibodies, with HER2-targeting antibodies such as trastuzumab, CD138-targeting antibodies such as B-B4, and EPHA2-targeting antibodies such as 1C1 providing particularly improved drug:antibody ratios, aggregation levels, stability (i.e., in vitro and in vivo stability), tumor targeting (i.e., cytotoxicity, efficacy), and / or therapeutic efficacy. Improved therapeutic efficacy can be measured in vitro or in vivo and may include a reduction in tumor growth rate and / or a reduction in tumor volume.

[0307] In certain embodiments, another antibody against the same target or an antibody against a different antigen target is used, resulting in at least some of the advantageous functional properties described above (e.g., improved stability, improved tumor targeting, improved therapeutic efficacy, etc.). In some embodiments, some or all of these advantageous functional properties are observed when the disclosed linkers and splicing modulator payloads are conjugated to another HER2-targeting, CD138-targeting, or EPHA2-targeting antibody or antigen-binding fragment. In some other embodiments, some or all of these advantageous functional properties are observed when the disclosed linkers and splicing modulator payloads are conjugated to a HER2-targeting antibody or antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment targets HER2. In some embodiments, the HER2-targeting antibody or antigen-binding fragment is trastuzumab. In some other embodiments, some or all of these advantageous functional properties are observed when the disclosed linkers and splicing modulator payloads are conjugated to a CD138-targeting antibody or antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment targets CD138. In some embodiments, the CD138-targeting antibody or antigen-binding fragment is B-B4. In some other embodiments, some or all of these advantageous functional properties are observed when the disclosed linkers and splicing modulator payloads are conjugated to an EPHA2-targeting antibody or antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment targets EPHA2. In some embodiments, the EPHA2-targeting antibody or antigen-binding fragment is 1C1.

[0308] Linker In various embodiments, the linker of an ADC is sufficiently stable extracellularly to be therapeutically effective. In some embodiments, the linker is stable outside of a cell, such that the ADC remains intact when present in extracellular conditions (e.g., before transport or delivery to a cell). The term "intact," as used in reference to an ADC, means that the antibody or antigen-binding fragment remains attached to the drug moiety (e.g., a splicing modulator). As used herein, "stable," in reference to a linker or an ADC comprising a linker, means that 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% (or any percentage therebetween) of the linkers present in a sample of the ADC are cleaved (or otherwise not intact in the case of the total ADC) when the ADC is present under extracellular conditions. In some embodiments, the linkers and / or ADCs disclosed herein are surprisingly stable compared to ADCs having alternative linkers and / or alternative linker and / or splicing modulator payloads. In some embodiments, the ADCs 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.

[0309] Whether a linker is stable extracellularly can be determined, for example, by placing the ADC in plasma for a predetermined period of time (e.g., 2, 4, 6, 8, 16, 24, 48, or 72 hours) and then quantifying the amount of free drug moiety present in the plasma. Stability may allow sufficient time for the ADC to localize to target tumor cells and prevent premature release of the drug moiety (which could reduce the therapeutic index of the ADC by indiscriminately damaging both normal and tumor tissue). In some embodiments, the linker is stable outside the target cell and, once inside the cell, releases the drug moiety from the ADC, allowing the drug to bind to its target (e.g., to the SF3b spliceosome complex). Thus, an effective linker will (i) maintain the specific binding properties of the antibody or antigen-binding fragment; (ii) enable delivery of the drug moiety, e.g., intracellular delivery, through stable association with the antibody or antigen-binding fragment; (iii) remain stable and intact until the ADC has been transported or delivered to its target site; and (iv) achieve the therapeutic effect, e.g., cytotoxic effect, of the drug moiety after cleavage or another release mechanism.

[0310] Linkers can affect the physicochemical properties of ADCs. Many cytotoxic agents are inherently hydrophobic, and linking them to antibodies with additional hydrophobic moieties can lead to aggregation. ADC aggregates are insoluble and often limit the drug loading achievable on the antibody, which can adversely affect ADC efficacy. Protein aggregates in biologics are also commonly associated with increased immunogenicity. As shown below, the linkers disclosed herein result in ADCs with low levels of aggregation and desirable drug loading levels.

[0311] 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 splicing modulator) when subjected to certain environmental factors, such as when internalized within a target cell, whereas non-cleavable linkers generally rely on degradation of the antibody or antigen-binding fragment itself.

[0312] In some embodiments, the linker is a non-cleavable linker. In some embodiments, the splicing modulator drug moiety of the ADC is released by degradation of the antibody or antigen-binding fragment. Non-cleavable linkers tend to remain covalently associated with at least one amino acid of the antibody and drug, even upon internalization by and degradation within the target cell. Numerous exemplary non-cleavable linkers are described herein, and others are known in the art. Exemplary non-cleavable linkers may include thioether, cyclohexyl, N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), or N-hydroxysuccinimide (NHS), one or more polyethylene glycol (PEG) moieties, e.g., 1, 2, 3, 4, 5, or 6 PEG moieties, or one or more alkyl moieties.

[0313] In some embodiments, the linker is a cleavable linker. A cleavable linker refers to any linker that includes a cleavable moiety. As used herein, the term "cleavable moiety" refers to any chemical bond that can be cleaved. Suitable cleavable chemical bonds are well known in the art and include, but are not limited to, acid-labile bonds, protease / peptidase-labile bonds, photolabile bonds, disulfide bonds, and esterase-labile bonds. A linker that includes a cleavable moiety may allow for release of the splicing modulator drug moiety from the ADC upon cleavage at a specific site in the linker.

[0314] In some embodiments, the linker is cleavable under intracellular conditions, such that cleavage of the linker is sufficient in the intracellular environment to release the splicing modulator drug moiety from the antibody or antigen-binding fragment and activate the drug and / or make the drug therapeutically effective. In some embodiments, the splicing modulator drug moiety is not cleaved from the antibody or antigen-binding fragment until the ADC is inside a cell that expresses an antigen specific for the antibody or antigen-binding fragment of the ADC, and upon entry into the cell, the splicing modulator drug moiety is cleaved from the antibody or antigen-binding fragment. In some embodiments, the linker comprises a cleavable moiety positioned such that upon cleavage, no portion of the linker or antibody or antigen-binding fragment remains attached to the splicing modulator drug moiety. Exemplary cleavable linkers include acid-labile linkers, protease / peptidase-sensitive linkers, photolabile linkers, dimethyl-containing, disulfide-containing, or sulfonamide-containing linkers.

[0315] In some embodiments, the linker is a pH-sensitive linker, which is sensitive to hydrolysis at a specific pH value. Typically, pH-sensitive linkers are cleavable under acidic conditions. This cleavage strategy generally takes advantage of the lower pH of intracellular compartments, such as endosomes (pH approximately 5-6) and lysosomes (pH approximately 4.8), compared to the cytosol (pH approximately 7.4), to induce hydrolysis of an acid-labile group in the linker, such as a hydrazone (Jain et al. (2015) Pharm Res 32:3526-40). In some embodiments, the linker is an acid-labile and / or hydrolyzable linker. For example, an acid-labile linker that is hydrolyzable in the lysosome and contains an acid-labile group (e.g., hydrazone, semicarbazone, thiosemicarbazone, cis-aconitic acid amide, orthoester, acetal, ketal, etc.) can be used. See, e.g., U.S. Patent Nos. 5,122,368; 5,824,805; 5,622,929; Dubowchik and Walker (1999) Pharm Therapeutics 83:67-123; Neville et al. (1989) Biol Chem. 264:14653-61. Such linkers are relatively stable under neutral pH conditions, such as those found in blood, but are unstable below pH 5.5 or 5.0, which approximates the pH of lysosomes. In certain embodiments, the hydrolyzable linker is a thioether linker (e.g., a thioether attached to a therapeutic agent via an acylhydrazone bond) (see, e.g., U.S. Patent No. 5,622,929).

[0316] In some embodiments, the linker is cleavable under reducing conditions. In some embodiments, the linker is cleavable in the presence of a reducing agent such as glutathione or dithiothreitol. In some embodiments, the linker is a cleavable disulfide linker or a cleavable sulfonamide linker.

[0317] In some embodiments, the linker is a cleavable disulfide linker. A variety of disulfide linkers are known in the art, including, for example, SATA (N-succinimidyl-5-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate), and SMPT (N-succinimidyloxycarbonyl-α-methyl-α-(2-pyridyldithio)toluene), including those that can be formed using SPDB and SMPT. See, e.g., Thorpe et al. (1987) Cancer Res. 47:5924-31; Wawrzynczak et al., In Immunoconjugates: Antibody Conjugates in Radioimagery and Therapy of Cancer (C.W. Vogel ed., Oxford U. Press, 1987). See also U.S. Pat. No. 4,880,935. Disulfide linkers are typically used to take advantage of the abundance of intracellular thiols that can promote cleavage of the disulfide bond. The intracellular concentration of the most abundant intracellular thiol, reduced glutathione, is generally in the 1-10 nM range, approximately 1,000-fold higher than the most abundant small thiol (i.e., cysteine) in blood, which is approximately 5 μM (Goldmacher et al., In Cancer Drug Discovery and Development: Antibody-Drug Conjugates and Immunotoxins (GLP Hillips ed., Springer, 2013)). Intracellular enzymes of the protein disulfide isomerase family can also contribute to the intracellular cleavage of disulfide linkers. As used herein, a cleavable disulfide linker refers to any linker that contains a cleavable disulfide moiety. The term "cleavable disulfide moiety" refers to a disulfide bond that can be cleaved and / or reduced, for example, by a thiol or an enzyme.

[0318] In some embodiments, the linker is a cleavable sulfonamide linker. As used herein, a cleavable sulfonamide linker refers to any linker that includes a cleavable sulfonamide moiety. The term "cleavable sulfonamide moiety" refers to a sulfonamide group, i.e., a sulfonyl group connected to an amine group, where the sulfur-nitrogen bond can be cleaved.

[0319] In some embodiments, the linker may be a dendritic linker for covalently linking two or more drug moieties to an antibody or antigen-binding fragment via a branched, multifunctional linker moiety. See, e.g., Sun et al. (2002) Bioorg Med Chem Lett. 12:2213-5; Sun et al. (2003) Bioorg Med Chem. 11:1761-8. Dendritic linkers can increase the drug-to-antibody molar ratio, i.e., drug loading, which is related to the efficacy of the ADC. Thus, for example, when an antibody or antigen-binding fragment bears only one reactive cysteine ​​thiol group, multiple splicing modulator drug moieties can be attached via a dendritic linker. In some embodiments, linker moieties or linker-drug moieties can be attached to an antibody or antigen-binding fragment via reduced disulfide crosslinking chemistry or lysine-specific techniques. See, for example, WO 2013 / 173391 and WO 2013 / 173393.

[0320] In some embodiments, the linker is cleavable by a cleaving agent, e.g., an enzyme, present in the intracellular environment (e.g., within a lysosome or endosome or caveolae). The linker can be, for example, a peptide linker that is cleaved by an intracellular peptidase or protease enzyme, including, but not limited to, a lysosomal or endosomal protease.

[0321] In some embodiments, the linker is a cleavable peptide linker. As used herein, a cleavable peptide linker refers to any linker comprising a cleavable peptide moiety. The term "cleavable peptide moiety" refers to any chemically linked amino acids (natural or synthetic amino acid derivatives) that can be cleaved by an agent present in the intracellular environment. For example, the linker may comprise a valine-alanine (Val-Ala) sequence or a valine-citrulline (Val-Cit) sequence that is cleavable by a peptidase such as a cathepsin, e.g., cathepsin B. In some embodiments, the linker may comprise a glutamic acid-valine-citrulline (Glu-Val-Cit) sequence. In some embodiments, the linker is an enzyme-cleavable linker, where the cleavable peptide moiety in the linker is cleavable by an enzyme. In some embodiments, the cleavable peptide moiety is cleavable by a lysosomal enzyme, e.g., a cathepsin. In some embodiments, the linker is a cathepsin-cleavable linker. In some embodiments, the cleavable peptide moiety in the linker is cleavable by a lysosomal cysteine ​​cathepsin, such as cathepsin B, C, F, H, K, L, O, S, V, X, or W. In some embodiments, the cleavable peptide moiety is cleavable by cathepsin B. An exemplary dipeptide that can be cleaved by cathepsin B is valine-citrulline (Val-Cit) (Dubowchik et al. (2002) Bioconjugate Chem. 13:855-69).

[0322] In some embodiments, the linker or cleavable peptide moiety in the linker comprises an amino acid unit. In some embodiments, the amino acid unit allows for cleavage of the linker by a protease, thus facilitating release of the splicing modulator drug moiety from the ADC upon exposure to one or more intracellular proteases, such as one or more lysosomal enzymes (Doronina et al. (2003) Nat Biotechnol. 21:778-84; Dubowchik and Walker (1999) Pharm Therapeutics 83:67-123). Exemplary amino acid units include, but are not limited to, dipeptides, tripeptides, tetrapeptides, and pentapeptides. Exemplary dipeptides include, but are not limited to, valine-alanine (Val-Ala), valine-citrulline (Val-Cit), alanine-asparagine (Ala-Asn), alanine-phenylalanine (Ala-Phe), phenylalanine-lysine (Phe-Lys), alanine-lysine (Ala-Lys), alanine-valine (Ala-Val), valine-lysine (Val-Lys), lysine-lysine (Lys-Lys), phenylalanine-citrulline (Phe-Cit), leucine-citrulline (Leu-Cit), isoleucine-citrulline (Ile-Cit), tryptophan-citrulline (Trp-Cit), and phenylalanine-alanine (Phe-Ala). Exemplary tripeptides include, but are not limited to, alanine-alanine-asparagine (Ala-Ala-Asn), glycine-valine-citrulline (Gly-Val-Cit), glycine-glycine-glycine (Gly-Gly-Gly), phenylalanine-phenylalanine-lysine (Phe-Phe-Lys), glutamic acid-valine-citrulline (Glu-Val-Cit) (see, e.g., Anami et al. (2018) Nat Comm. 9:2512, which is incorporated herein by reference for exemplary linkers including Glu-Val-Cit), and glycine-phenylalanine-lysine (Gly-Phe-Lys).Other exemplary amino acid units include, but are not limited to, Gly-Phe-Gly-Gly (SEQ ID NO: 34), Gly-Phe-Leu-Gly (SEQ ID NO: 35), Ala-Leu-Ala-Leu (SEQ ID NO: 36), Phe-N, as described, for example, in U.S. Pat. No. 6,214,345. 9 -Tosyl-Arg, and Phe-N 9 In some embodiments, the amino acid unit in the linker comprises Val-Ala. In some embodiments, the amino acid unit in the linker comprises Val-Cit. In some embodiments, the amino acid unit in the linker comprises Glu-Val-Cit. The amino acid unit may comprise naturally occurring amino acid residues and / or minor amino acids and / or non-naturally occurring amino acid analogs, such as citrulline. The amino acid unit may be designed and optimized for enzymatic cleavage by specific enzymes, such as tumor-associated proteases, lysosomal proteases, such as cathepsin B, C, D, or S, or plasmin proteases.

[0323] In some embodiments, the linker is a cleavable β-glucuronide linker. As used herein, a cleavable β-glucuronide linker refers to any linker that includes a cleavable β-glucuronide moiety. An exemplary cleavable β-glucuronide linker has the structure: [ka] Includes.

[0324] The term "cleavable β-glucuronide moiety" refers to a glycosidic bond that can be cleaved by an agent having β-glucuronidase activity. In some embodiments, the linker comprises a glycosidic bond that can be cleaved by β-glucuronidase. β-glucuronidase is a UDP-glucuronosyltransferase that catalyzes the hydrolysis of glycosidic bonds in glucuronides having a β-configuration.

[0325] In some embodiments, the ADCs disclosed herein include a cleavable β-glucuronide moiety in the linker that is cleavable by an enzyme. In some embodiments, the cleavable β-glucuronide moiety in the linker is cleavable by a lysosomal enzyme, such as β-glucuronidase. In some embodiments, the linker is a β-glucuronidase-cleavable linker. In some embodiments, the cleavable β-glucuronide moiety in the linker allows cleavage of the linker by β-glucuronidase after internalization of the ADC, thereby facilitating release of the drug moiety from the ADC in the cellular environment.

[0326] In some embodiments, the linker in any ADC disclosed herein may comprise at least one spacer unit that links the antibody or antigen-binding fragment to the drug moiety (e.g., a splicing modulator drug moiety). In some embodiments, the spacer unit between the antibody or antigen-binding fragment and the cleavable moiety, if present, links the cleavage site (e.g., a cleavable peptide moiety) in the linker to the antibody or antigen-binding fragment. In some embodiments, the spacer unit between the drug moiety and the cleavable moiety, if present, links the cleavage site (e.g., a cleavable peptide moiety) in the linker to the drug moiety. In some embodiments, the cleavage site is not present and a spacer unit is used to link the antibody or antigen-binding fragment to the drug moiety.

[0327] In some embodiments, the linker and / or the spacer unit in the linker are substantially hydrophilic. The use of a hydrophilic linker can reduce the extent to which the drug can be pumped out of resistant cancer cells by multidrug resistance (MDR) or functionally similar transporters. In some embodiments, the hydrophilic linker can include one or more polyethylene glycol (PEG) moieties, for example, 1, 2, 3, 4, 5, or 6 PEG moieties. In some embodiments, the linker includes two PEG moieties.

[0328] In some embodiments, the spacer unit in the linker comprises one or more PEG moieties. In some embodiments, the spacer unit comprises one or more -(PEG) m -, and m is an integer from 1 to 10 (i.e., m can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, m ranges from 1 to 10; 2 to 8; 2 to 6; 2 to 5; 2 to 4; or 2 to 3. In some embodiments, m is 2. In some embodiments, the spacer unit is (PEG)2, (PEG)3, (PEG)4, (PEG)5, (PEG)6, (PEG)7, (PEG)8, (PEG)9, or (PEG) 10 In some embodiments, the spacer unit comprises (PEG).

[0329] In some embodiments, the spacer unit in the linker comprises an alkyl moiety. In some embodiments, the spacer unit comprises one or more -(CH) n -, where n is an integer between 1 and 10 (i.e., n can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, n ranges from 1 to 10; 2 to 8; 2 to 6; 2 to 5; 2 to 4; or 2 to 3. In some embodiments, n is 2. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, the spacer unit is (CH2)2, (CH2)3, (CH2)4, (CH2)5, (CH2)6, (CH2)7, (CH2)8, (CH2)9, or (CH2) 10 In some embodiments, the spacer unit comprises (CH2)2 ("Et"). In some embodiments, the spacer unit comprises (CH2)6 ("Hex"). In some embodiments, the spacer unit comprises (CH2)2-O-(CH2)2 ("Et-O-Et").

[0330] The spacer unit can be used, for example, to link an antibody or antigen-binding fragment to a drug moiety directly or indirectly. In some embodiments, the spacer unit directly links the antibody or antigen-binding fragment to the splicing modulator drug moiety. In some embodiments, the antibody or antigen-binding fragment and the splicing modulator drug moiety are linked via a spacer unit comprising one or more PEG moieties (e.g., (PEG)2) or one or more alkyl moieties (e.g., (CH2)2, (CH2)6, or (CH2)2-O-(CH2)2). In some embodiments, the spacer unit indirectly links the antibody or antigen-binding fragment to the splicing modulator drug moiety. In some embodiments, the spacer unit indirectly links the antibody or antigen-binding fragment to the splicing modulator drug moiety through a cleavable moiety (e.g., a cleavable peptide or a cleavable β-glucuronide) and / or a linking moiety, e.g., a maleimide moiety, for joining the spacer unit to the antibody or antigen-binding fragment.

[0331] The spacer unit, in various embodiments, is attached to the antibody or antigen-binding fragment (ie, antibody or antigen-binding fragment) via a maleimide (Mal) moiety.

[0332] A spacer unit that is attached to an antibody or antigen-binding fragment via a Mal is referred to herein as a "Mal-spacer unit." The term "Mal" or "maleimide moiety," as used herein, refers to a compound that contains a maleimide group and is reactive with sulfhydryl groups, such as the sulfhydryl groups of cysteine ​​residues on an antibody or antigen-binding fragment. Other functional groups reactive with sulfhydryl groups (thiols) include, but are not limited to, iodoacetamide, bromoacetamide, vinylpyridine, disulfide, pyridyl disulfide, isocyanate, and isothiocyanate. In some embodiments, a Mal-spacer unit is reactive with cysteine ​​residues on an antibody or antigen-binding fragment. In some embodiments, a Mal-spacer unit is attached to an antibody or antigen-binding fragment via a cysteine ​​residue. In some embodiments, a Mal-spacer unit comprises a PEG moiety. In some embodiments, a Mal-spacer unit comprises an alkyl moiety.

[0333] In certain embodiments, the linker comprises a Mal-spacer unit and a cleavable peptide moiety. In some embodiments, the cleavable peptide moiety comprises an amino acid unit. In some embodiments, the amino acid unit comprises Val-Cit. In some embodiments, the amino acid unit comprises Val-Ala. In some embodiments, the amino acid unit comprises Glu-Val-Cit. In some embodiments, the linker comprises a Mal-spacer unit and Val-Cit. In some embodiments, the linker comprises a Mal-spacer unit and Val-Ala. In some embodiments, the linker comprises a Mal-spacer unit and Val-Cit, wherein the Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the linker comprises a Mal-spacer unit and Val-Ala, wherein the Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the linker comprises a Mal-spacer unit and a cleavable β-glucuronide moiety.

[0334] In some embodiments, the linker comprises the structure: Mal-spacer unit. In some embodiments, the Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the linker comprises the structure: MC. In some embodiments, the linker comprises the structure: Mal-(CH2)2 ("Mal-Et"). In some embodiments, the linker comprises the structure: Mal-(CH2)6 ("Mal-Hex"). In some embodiments, the linker comprises the structure: Mal-(CH2)2-O-(CH2)2 ("Mal-Et-O-Et"). In some embodiments, the linker comprises the structure: Mal-(PEG)2. In some embodiments, the linker comprises the structure: Mal-(PEG)2-CO.

[0335] In various embodiments, a Mal-spacer unit links the antibody or antigen-binding fragment to the cleavable peptide moiety. In some embodiments, the linker comprises Mal-spacer unit-peptide. In some embodiments, the linker comprises the structure: Mal-spacer unit-Val-Cit. In some embodiments, the Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the linker comprises the structure: MC-Val-Cit.

[0336] In some embodiments, the linker comprises the structure: Mal-spacer unit-Val-Ala. In some embodiments, the Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the linker comprises the structure: MC-Val-Ala.

[0337] In various embodiments, a Mal-spacer unit links the antibody or antigen-binding fragment to a cleavable β-glucuronide moiety. In some embodiments, the linker comprises a Mal-spacer unit-β-glucuronide. In some embodiments, the linker comprises an MC-β-glucuronide.

[0338] In various embodiments, the cleavable portion of the linker is directly joined to the splicing modulator drug moiety. In other embodiments, a spacer unit is used to connect the cleavable portion of the linker to the splicing modulator drug moiety. In various embodiments, the splicing modulator is connected to the cleavable portion of the linker by a spacer unit.

[0339] A spacer unit can be "self-immolative" or "non-self-immolative." A "non-self-immolative" spacer unit is one in which some or all of the spacer unit remains attached to the splicing modulator drug moiety upon cleavage of the linker. Examples of non-self-immolative spacer units include, but are not limited to, a glycine spacer unit and a glycine-glycine spacer unit. A non-self-immolative spacer unit may eventually degrade over time, but does not immediately release the entire linked native drug moiety, even under cellular conditions. A "self-immolative" spacer unit allows for release of the native drug moiety under intracellular conditions. A "native drug" or "native drug moiety" is one in which no part of the spacer unit or other chemical modification remains after cleavage / degradation of the spacer unit.

[0340] Self-immolative chemistries are known in the art and can be readily selected for the disclosed ADCs. In various embodiments, the spacer unit connecting the cleavable portion of the linker to the splicing modulator drug moiety is self-immolative and undergoes self-immolation simultaneously with or immediately before / after cleavage of the cleavable portion under intracellular conditions. In some embodiments, the splicing modulator is connected to the cleavable portion of the linker by a self-immolative spacer unit. In certain embodiments, the splicing modulator is connected to the cleavable portion of the linker by a self-immolative spacer unit, the cleavable portion comprises Val-Cit, and maleimidocaproyl (MC) joins the cleavable portion to the antibody or antigen-binding fragment. In certain embodiments, the splicing modulator is connected to the cleavable portion of the linker by a self-immolative spacer unit, the cleavable portion comprises Val-Ala, and maleimidocaproyl (MC) joins the cleavable portion to the antibody or antigen-binding fragment. In certain embodiments, the splicing modulator is linked to the cleavable portion of the linker by a self-immolative spacer unit, where the cleavable portion comprises Glu-Val-Cit, and maleimidocaproyl (MC) joins the cleavable portion to the antibody or antigen-binding fragment. In certain embodiments, the splicing modulator is joined to the antibody or antigen-binding fragment via a Mal-spacer unit (e.g., MC) in the linker joined to a pABC or pAB self-immolative spacer unit and a Val-Cit cleavable portion. In certain other embodiments, the splicing modulator is joined to the antibody or antigen-binding fragment via a Mal-spacer unit (e.g., MC) in the linker joined to a pABC or pAB self-immolative spacer unit and a Val-Ala cleavable portion. In certain other embodiments, the splicing modulator is conjugated to the antibody or antigen-binding fragment via a Mal-spacer unit (e.g., MC) in a linker joined to a pABC or pAB self-immolative spacer unit and a Glu-Val-Cit cleavable moiety.

[0341] In certain embodiments, the self-immolative spacer unit in the linker comprises a p-aminobenzyl unit. In some embodiments, p-aminobenzyl alcohol (pABOH) is attached to the amino acid unit or other cleavable moiety in the linker via an amide bond, creating a carbamate, methylcarbamate, or carbonate between pABOH and the drug moiety (Hamann et al. (2005) Expert Opin Ther Patents 15:1087-103). In some embodiments, the self-immolative spacer unit is or includes p-aminobenzyloxycarbonyl (pABC). Without being bound by theory, the self-immolation of pABC is believed to involve a spontaneous 1,6-elimination reaction (Jain et al. (2015) Pharm Res. 32:3526-40).

[0342] In various embodiments, the structure of p-aminobenzyloxycarbonyl (pABC) used in the disclosed ADCs is shown below: [ka]

[0343] In various embodiments, a self-immolative spacer unit links the cleavable portion of the linker to the splicing modulator. In some embodiments, the self-immolative spacer unit is pABC. In some embodiments, pABC links the cleavable portion of the linker to the splicing modulator. In some embodiments, pABC undergoes self-immolation upon cleavage of the cleavable portion, and the splicing modulator is released from the ADC in its native, active form.

[0344] In some embodiments, the anti-HER2 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Cit-pABC, hi other embodiments, the anti-HER2 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Ala-pABC.

[0345] In some embodiments, the anti-CD138 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Cit-pABC, hi other embodiments, the anti-CD138 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Ala-pABC.

[0346] In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Cit-pABC, hi other embodiments, the anti-EPHA2 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Ala-pABC.

[0347] In some embodiments, pABC undergoes self-immolation upon cleavage of the cleavable peptide moiety in the linker. In some embodiments, the cleavable peptide moiety comprises an amino acid unit. In some embodiments, the linker comprises the amino acid unit-pABC. In some embodiments, the amino acid unit is Val-Cit. In some embodiments, the linker comprises Val-Cit-pABC. In some embodiments, the amino acid unit is Val-Ala. In some embodiments, the linker comprises Val-Ala-pABC. In some embodiments, the amino acid unit is Glu-Val-Cit. In some embodiments, the linker comprises Glu-Val-Cit-pABC. In some embodiments, the amino acid unit is Ala-Ala-Asn. In some embodiments, the linker comprises Ala-Ala-Asn-pABC.

[0348] In some embodiments, pABC undergoes self-immolation upon cleavage of the cleavable β-glucuronide moiety in the linker, hi some embodiments, the linker comprises β-glucuronide-pABC.

[0349] In certain embodiments, the self-immolative spacer unit in the linker comprises a p-aminobenzyl unit. In some embodiments, the self-immolative spacer unit in the linker comprises p-aminobenzyl (pAB). In some embodiments, the self-immolation of pAB involves a spontaneous 1,6-elimination reaction.

[0350] In various embodiments, the structure of p-aminobenzyl (pAB) used in the disclosed ADCs is shown below: [ka]

[0351] In various embodiments, a self-immolative spacer unit links the cleavable portion of the linker to the splicing modulator. In some embodiments, the self-immolative spacer unit is pAB. In some embodiments, pAB links the cleavable portion of the linker to the splicing modulator. In some embodiments, pAB undergoes self-immolation upon cleavage of the cleavable portion, releasing the splicing modulator from the ADC in its native, active form.

[0352] In some embodiments, the anti-HER2 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Cit-pAB, hi other embodiments, the anti-HER2 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Ala-pAB.

[0353] In some embodiments, the anti-CD138 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Cit-pAB, hi other embodiments, the anti-CD138 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Ala-pAB.

[0354] In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Cit-pAB, hi other embodiments, the anti-EPHA2 antibody or antigen-binding fragment is tethered to the splicing modulator by a linker comprising MC-Val-Ala-pAB.

[0355] In some embodiments, pAB undergoes self-immolation upon cleavage of the cleavable peptide moiety in the linker. In some embodiments, the cleavable peptide moiety comprises an amino acid unit. In some embodiments, the linker comprises the amino acid unit -pAB. In some embodiments, the amino acid unit is Val-Cit. In some embodiments, the linker comprises Val-Cit-pAB. In some embodiments, the amino acid unit is Val-Ala. In some embodiments, the linker comprises Val-Ala-pAB. In some embodiments, the amino acid unit is Glu-Val-Cit. In some embodiments, the linker comprises Glu-Val-Cit-pAB. In some embodiments, the amino acid unit is Ala-Ala-Asn. In some embodiments, the linker comprises Ala-Ala-Asn-pAB.

[0356] In some embodiments, pAB undergoes self-immolation upon cleavage of the cleavable β-glucuronide moiety in the linker, hi some embodiments, the linker comprises β-glucuronide-pAB.

[0357] In some other embodiments, the splicing modulator is linked to the cleavable portion of the linker by a non-self-immolative spacer unit. In certain embodiments, the splicing modulator is linked to the cleavable portion of the linker by a non-self-immolative spacer unit, the cleavable portion comprises Val-Cit, and maleimidocaproyl (MC) joins the cleavable portion to the antibody or antigen-binding fragment. In certain embodiments, the splicing modulator is linked to the cleavable portion of the linker by a non-self-immolative spacer unit, the cleavable portion comprises Val-Ala, and maleimidocaproyl (MC) joins the cleavable portion to the antibody or antigen-binding fragment.

[0358] In various aspects, the antibody or antigen-binding fragment of the ADC is conjugated to the splicing modulator drug moiety via a linker, wherein the linker comprises a Mal-spacer unit (e.g., MC), a cleavable amino acid unit, and pABC. In some embodiments, the spacer unit comprises an alkyl moiety. In some embodiments, the Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the linker comprises Mal-spacer unit-amino acid unit-pABC. In some embodiments, the linker comprises MC-amino acid unit-pABC. In some embodiments, the linker comprises MC-Val-Cit-pABC. In some embodiments, the linker comprises MC-Val-Ala-pABC. In some embodiments, the linker comprises MC-Glu-Val-Cit-pABC. In some embodiments, the linker comprises MC-Ala-Ala-Asn-pABC.

[0359] In various other aspects, the antibody or antigen-binding fragment of the ADC is conjugated to the splicing modulator drug moiety via a linker, wherein the linker comprises a Mal-spacer unit (e.g., MC), a cleavable amino acid unit, and pAB. In some embodiments, the spacer unit comprises an alkyl moiety. In some embodiments, the Mal-spacer unit comprises maleimidocaproyl (MC). In some embodiments, the linker comprises Mal-spacer unit-amino acid unit-pAB. In some embodiments, the linker comprises MC-amino acid unit-pAB. In some embodiments, the linker comprises MC-Val-Cit-pAB. In some embodiments, the linker comprises MC-Val-Ala-pAB. In some embodiments, the linker comprises MC-Glu-Val-Cit-pAB. In some embodiments, the linker comprises MC-Ala-Ala-Asn-pAB.

[0360] In various other aspects, the antibody or antigen-binding fragment of the ADC is conjugated to the splicing modulator drug moiety via a linker, where the linker comprises a Mal-spacer unit (e.g., MC), a cleavable β-glucuronide, and pABC. In some embodiments, the linker comprises Mal-spacer unit-β-glucuronide-pABC. In some embodiments, the linker comprises MC-β-glucuronide-pABC.

[0361] In yet other aspects, the antibody or antigen-binding fragment of the ADC is conjugated to the splicing modulator drug moiety via a linker, where the linker comprises a Mal-spacer unit (e.g., MC), a cleavable β-glucuronide, and pAB. In some embodiments, the linker comprises Mal-spacer unit-β-glucuronide-pAB. In some embodiments, the linker comprises MC-β-glucuronide-pAB.

[0362] In various embodiments, the ADC compound has formula (I): Ab-(LD) p (I) where Ab is an antibody or antigen-binding fragment that targets a neoplastic cell; D is a splicing modulator; L is a linker covalently linking Ab to D; and p is an integer from 1 to 15. It has.

[0363] In some embodiments, the antibody or antigen-binding fragment (Ab) of the ADC is conjugated to the splicing modulator drug moiety via a linker, where the linker is any of the linkers disclosed herein or incorporated by reference, or comprises one or more components of any of the linkers disclosed herein or incorporated by reference.

[0364] In some embodiments, the linker comprises a cleavable moiety positioned such that after cleavage, no portion of the linker or antibody or antigen-binding fragment remains attached to the splicing modulator. In some embodiments, the cleavable moiety is a cleavable peptide moiety, e.g., an amino acid unit such as Val-Cit or Val-Ala. In some embodiments, the amino acid unit or linker comprises Val-Cit. In some embodiments, the amino acid unit or linker comprises Val-Ala. In some embodiments, the amino acid unit or linker comprises Glu-Val-Cit.

[0365] In some embodiments, the linker comprises at least one spacer unit that joins the antibody or antigen-binding fragment to the cleavable moiety. In some embodiments, the linker comprises at least one spacer unit that joins the antibody or antigen-binding fragment to the drug moiety. In some embodiments, the spacer unit or linker comprises at least one alkyl moiety.

[0366] In some embodiments, the spacer unit in the linker is attached to the antibody or antigen-binding fragment via a Mal moiety ("Mal-spacer unit"). In some embodiments, the Mal-spacer unit comprises at least one alkyl moiety. In some embodiments, the linker comprises maleimidocaproyl (MC). In some embodiments, the linker comprises Mal-(CH2)2 ("Mal-Et"). In some embodiments, the linker comprises Mal-(CH2)6 ("Mal-Hex"). In some embodiments, the linker comprises Mal-(CH2)2-O-(CH2)2 ("Mal-Et-O-Et"). In some embodiments, the linker comprises Mal-(PEG)2-CO. In some embodiments, the Mal-spacer unit attaches the antibody or antigen-binding fragment to the drug moiety.

[0367] In some embodiments, the Mal-spacer unit or linker comprises Mal-(PEG)2, Mal-(PEG)3, Mal-(PEG)4, Mal-(PEG)5, Mal-(PEG)6, Mal-(PEG)7, or Mal-(PEG)8. In some embodiments, the Mal-spacer unit or linker comprises Mal-(PEG)2. In some embodiments, the Mal-spacer unit or linker comprises Mal-(PEG)2-CO, Mal-(PEG)3-CO, Mal-(PEG)4-CO, Mal-(PEG)5-CO, Mal-(PEG)6-CO, Mal-(PEG)7-CO, or Mal-(PEG)8-CO. In some embodiments, the Mal-spacer unit or linker comprises Mal-(PEG)2-CO. In some embodiments, the Mal-spacer unit or linker comprises Mal-(PEG)2-CO and at least one additional spacer unit. In some embodiments, Mal-(PEG)2-CO links the antibody or antigen-binding fragment to the drug moiety. In some embodiments, the linker comprises or consists of Mal-(PEG)2-CO. An example of a "Mal-(PEG)2-CO" linker is also referred to herein as an "ADL2" or "ADL2" linker.

[0368] In some embodiments, the Mal-spacer unit or linker comprises MC. In some embodiments, the Mal-spacer unit or linker comprises MC and at least one additional spacer unit. In some embodiments, the MC links the antibody or antigen-binding fragment to the drug moiety. In some embodiments, the linker comprises or consists of MC. Examples of "MC" linkers are also referred to herein as "ADL10" or "ADL10" linkers.

[0369] In some embodiments, the Mal-spacer unit or linker comprises Mal-(CH2)6 ("Mal-Hex"). In some embodiments, the Mal-spacer unit or linker comprises Mal-Hex and at least one additional spacer unit. In some embodiments, the Mal-Hex links the antibody or antigen-binding fragment to the drug moiety. In some embodiments, the linker comprises Mal-Hex. Examples of "Mal-Hex" linkers are also referred to herein as "ADL12" or "ADL12" linkers.

[0370] In some embodiments, the Mal-spacer unit or linker comprises Mal-(CH2)2 ("Mal-Et"). In some embodiments, the Mal-spacer unit or linker comprises Mal-Et and at least one additional spacer unit. In some embodiments, Mal-Et links the antibody or antigen-binding fragment to the drug moiety. In some embodiments, the linker comprises Mal-Et. An example of a "Mal-Et" linker is also referred to herein as an "ADL14" or "ADL14" linker.

[0371] In some embodiments, the Mal-spacer unit or linker comprises Mal-(CH2)2-O-(CH2)2 ("Mal-Et-O-Et"). In some embodiments, the Mal-spacer unit or linker comprises Mal-Et-O-Et and at least one additional spacer unit. In some embodiments, Mal-Et-O-Et connects the antibody or antigen-binding fragment to the drug moiety. In some embodiments, the linker comprises Mal-Et-O-Et. An example of a "Mal-Et-O-Et" linker is also referred to herein as an "ADL15" or "ADL15" linker.

[0372] In some other embodiments, a Mal-spacer unit links the antibody or antigen-binding fragment to the cleavable portion of the linker. In some embodiments, the cleavable portion of the linker is a cleavable peptide moiety, such as an amino acid unit. In some embodiments, the cleavable peptide moiety is Val-Cit or Val-Ala. In some embodiments, the Mal-spacer unit or linker comprises MC. In some embodiments, the linker comprises MC-Val-Cit. In some embodiments, the linker comprises MC-Val-Ala. In some embodiments, the linker comprises MC-Glu-Val-Cit. In some embodiments, the linker comprises MC-Ala-Ala-Asn.

[0373] In some embodiments, a spacer unit joins the cleavable moiety of the linker to the splicing modulator, hi some embodiments, the spacer unit joining the cleavable moiety to the splicing modulator is self-immolative.

[0374] In some embodiments, the spacer unit comprises pABC. In some embodiments, pABC links the cleavable moiety to the splicing modulator. In some embodiments, the cleavable moiety is a cleavable peptide moiety, such as an amino acid unit. In some embodiments, the linker comprises the amino acid unit-pABC.

[0375] In some embodiments, the linker comprises Val-Cit-pABC. In some embodiments, the linker comprises Val-Cit-pABC and an MC Mal-spacer unit that joins the linker to the antibody or antigen-binding fragment. In some embodiments, the linker comprises MC-Val-Cit-pABC. In some embodiments, the linker comprises MC-Val-Cit-pABC and at least one additional spacer unit. An example of an MC-Val-Cit-pABC linker is also referred to herein as an "ADL1" or "ADL1" linker.

[0376] In some embodiments, the linker comprises Val-Ala-pABC. In some embodiments, the linker comprises Val-Ala-pABC and an MC Mal-spacer unit that joins the linker to the antibody or antigen-binding fragment. In some embodiments, the linker comprises MC-Val-Ala-pABC. In some embodiments, the linker comprises MC-Val-Ala-pABC and at least one additional spacer unit. An example of an MC-Val-Ala-pABC linker is also referred to herein as an "ADL6" or "ADL6" linker.

[0377] In some embodiments, the linker comprises Glu-Val-Cit-pABC. In some embodiments, the linker comprises Glu-Val-Cit-pABC and an MC Mal-spacer unit that joins the linker to the antibody or antigen-binding fragment. In some embodiments, the linker comprises MC-Glu-Val-Cit-pABC. In some embodiments, the linker comprises MC-Glu-Val-Cit-pABC and at least one additional spacer unit. An example of an MC-Glu-Val-Cit-pABC linker is also referred to herein as an "ADL23" or "ADL23" linker.

[0378] In some embodiments, the linker comprises Ala-Ala-Asn-pABC. In some embodiments, the linker comprises Ala-Ala-Asn-pABC and an MC Mal-spacer unit that joins the linker to the antibody or antigen-binding fragment. In some embodiments, the linker comprises MC-Ala-Ala-Asn-pABC. In some embodiments, the linker comprises MC-Ala-Ala-Asn-pABC and at least one additional spacer unit. An example of an MC-Ala-Ala-Asn-pABC linker is also referred to herein as an "ADL21" or "ADL21" linker.

[0379] In some other embodiments, the spacer unit comprises pAB. In some embodiments, pAB links the cleavable moiety to the splicing modulator. In some embodiments, the cleavable moiety is a cleavable peptide moiety, such as an amino acid unit. In some embodiments, the linker comprises the amino acid unit -pAB.

[0380] In some embodiments, the linker comprises Val-Ala-pAB. In some embodiments, the linker comprises Val-Ala-pAB and an MC Mal-spacer unit that joins the linker to the antibody or antigen-binding fragment. In some embodiments, the linker comprises MC-Val-Ala-pAB. In some embodiments, the linker comprises MC-Val-Ala-pAB and at least one additional spacer unit. An example of an MC-Val-Ala-pAB linker is also referred to herein as an "ADL5" or "ADL5" linker.

[0381] In some embodiments, the linker comprises Val-Cit-pAB. In some embodiments, the linker comprises Val-Cit-pAB and an MC Mal-spacer unit that joins the linker to the antibody or antigen-binding fragment. In some embodiments, the linker comprises MC-Val-Cit-pAB. In some embodiments, the linker comprises MC-Val-Cit-pAB and at least one additional spacer unit. An example of an MC-Val-Cit-pAB linker is also referred to herein as an "ADL7" or "ADL7" linker.

[0382] In some embodiments, the linker comprises β-glucuronide-pABC. In some embodiments, the linker comprises β-glucuronide-pABC and an MC Mal-spacer unit that joins the linker to the antibody or antigen-binding fragment. In some embodiments, the linker comprises MC-β-glucuronide-pABC. In some embodiments, the linker comprises MC-β-glucuronide-pABC and at least one additional spacer unit. An example of MC-β-glucuronide-pABC is also referred to herein as "ADL13" or the "ADL13" linker.

[0383] In some embodiments, the linker comprises β-glucuronide-pAB. In some embodiments, the linker comprises β-glucuronide-pAB and an MC Mal-spacer unit that joins the linker to the antibody or antigen-binding fragment. In some embodiments, the linker comprises MC-β-glucuronide-pAB.

[0384] In some embodiments, the antibody or antigen-binding fragment is conjugated to the splicing modulator drug moiety via an ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15 linker. In various embodiments, ADCs comprising an ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15 linker disclosed herein and a splicing modulator drug moiety have been discovered to demonstrate desirable properties for therapeutic ADCs. In various embodiments, these properties include, but are not limited to, effective drug loading levels, low aggregation levels, stability under storage conditions or while circulating in the body (e.g., serum stability), retention of affinity for target-expressing cells comparable to that of an unconjugated antibody, potent cytotoxicity against target-expressing cells, low levels of off-target cell killing, high levels of bystander killing, and / or effective in vivo anti-cancer activity, all when compared to ADCs that use other linker-payloads. For example, in various embodiments, ADCs comprising an ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15 linker disclosed herein and a splicing modulator drug moiety exhibit increased ability to inhibit the growth and / or proliferation of target-expressing cells when compared to ADCs that use other linker-payloads (e.g., an ADL10 linker and a splicing modulator drug moiety). In various embodiments, ADCs comprising an ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15 linker disclosed herein and a splicing modulator drug moiety unexpectedly exhibit increased in vivo stability (e.g., plasma stability) when compared to other splicing modulator-based ADCs (e.g., tylanstatin A-based ADCs, e.g., as reported in Puthenveetil et al. Bioconjugate Chem. (2016) 27:1880-8).

[0385] In some embodiments, good or superior functional properties afforded by particular combinations of ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15 linkers disclosed herein with splicing modulator drug moieties may be observed in linker-payloads conjugated to, for example, anti-HER2 antibodies such as trastuzumab; anti-CD138 antibodies such as B-B4; or anti-EPHA2 antibodies such as 1C1.

[0386] In some embodiments, an ADC comprises an ADL1-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells. In some embodiments, an ADC comprises an ADL2-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells. In some embodiments, an ADC comprises an ADL5-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells. In some embodiments, an ADC comprises an ADL6-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells. In some embodiments, an ADC comprises an ADL7-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells. In some embodiments, an ADC comprises an ADL12-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells. In some embodiments, an ADC comprises an ADL13-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells. In some embodiments, an ADC comprises an ADL14-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells. In some embodiments, an ADC comprises an ADL15-splice modulator and an antibody or antigen-binding fragment thereof, the antibody or antigen-binding fragment thereof retaining the ability to target and internalize into neoplastic cells.

[0387] In some embodiments, an ADC comprises an ADL1-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2. In some embodiments, an ADC comprises an ADL2-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2. In some embodiments, an ADC comprises an ADL5-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2. In some embodiments, an ADC comprises an ADL6-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2. In some embodiments, an ADC comprises an ADL7-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2. In some embodiments, an ADC comprises an ADL12-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2. In some embodiments, an ADC comprises an ADL13-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2. In some embodiments, an ADC comprises an ADL14-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2. In some embodiments, an ADC comprises an ADL15-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing HER2.

[0388] In some embodiments, the antibody or antigen-binding fragment thereof that targets HER2-expressing neoplastic cells is an internalizing antibody or internalizing antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment thereof that targets HER2-expressing neoplastic cells comprises three heavy chain complementarity-determining regions (HCDRs) comprising the amino acid sequences of SEQ ID NO:1 (HCDR1), SEQ ID NO:2 (HCDR2), and SEQ ID NO:3 (HCDR3); and three light chain complementarity-determining regions (LCDRs) comprising the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[0389] In some embodiments, the ADC has formula (I): Ab-(LD) p (I) (In the formula, (i) the Ab is an anti-HER2 antibody or antigen-binding fragment thereof comprising three heavy chain complementarity-determining regions (HCDRs) comprising the amino acid sequences of SEQ ID NO:1 (HCDR1), SEQ ID NO:2 (HCDR2), and SEQ ID NO:3 (HCDR3); and three light chain complementarity-determining regions (LCDRs) comprising the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3); (ii) D is a splicing modulator; (iii) L is a linker comprising ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15; and (iv) p is an integer from 1 to 15. It has.

[0390] In some embodiments, an antibody or antigen-binding fragment thereof that targets HER2-expressing neoplastic cells comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, an antibody or antigen-binding fragment thereof that targets HER2-expressing neoplastic cells comprises a human IgG1 heavy chain constant domain and a human Ig kappa light chain constant domain. In some embodiments, the antibody is trastuzumab. In some embodiments, p is an integer from 1 to 10, 2 to 8, or 4 to 8. In some embodiments, p is 4. In some embodiments, p is 8.

[0391] In some embodiments, an ADC comprises an ADL1-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138. In some embodiments, an ADC comprises an ADL2-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138. In some embodiments, an ADC comprises an ADL5-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138. In some embodiments, an ADC comprises an ADL6-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138. In some embodiments, an ADC comprises an ADL7-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138. In some embodiments, an ADC comprises an ADL12-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138. In some embodiments, an ADC comprises an ADL13-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138. In some embodiments, an ADC comprises an ADL14-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138. In some embodiments, an ADC comprises an ADL15-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138.

[0392] In some embodiments, the antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138 is an internalizing antibody or internalizing antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138 comprises 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:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

[0393] In some embodiments, the ADC has formula (I): Ab-(LD) p (I) (In the formula, (i) the Ab is an anti-CD138 antibody or antigen-binding fragment thereof comprising 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:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3); (ii) D is a splicing modulator; (iii) L is a linker comprising ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15; and (iv) p is an integer from 1 to 15. It has.

[0394] In some embodiments, an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138 comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 22. In some embodiments, an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138 comprises a mouse IgG2a heavy chain constant domain and a mouse Ig kappa light chain constant domain. In some embodiments, an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing CD138 comprises a human IgG2a heavy chain constant domain and a human Ig kappa light chain constant domain. In some embodiments, the antibody is B-B4. In some embodiments, p is an integer between 1 and 10, between 2 and 8, or between 4 and 8. In some embodiments, p is 4. In some embodiments, p is 8.

[0395] In some embodiments, an ADC comprises an ADL1-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2. In some embodiments, an ADC comprises an ADL2-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2. In some embodiments, an ADC comprises an ADL5-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2. In some embodiments, an ADC comprises an ADL6-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2. In some embodiments, an ADC comprises an ADL7-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2. In some embodiments, an ADC comprises an ADL12-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2. In some embodiments, an ADC comprises an ADL13-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2. In some embodiments, an ADC comprises an ADL14-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2. In some embodiments, an ADC comprises an ADL15-splice modulator and an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2.

[0396] In some embodiments, the antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2 is an internalizing antibody or internalizing antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2 comprises three heavy chain complementarity-determining regions (HCDRs) comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3); and three light chain complementarity-determining regions (LCDRs) comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3).

[0397] In some embodiments, the ADC has formula (I): Ab-(LD) p (I) (In the formula, (i) the Ab is an anti-EPHA2 antibody or antigen-binding fragment thereof comprising three heavy chain complementarity-determining regions (HCDRs) comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3); and three light chain complementarity-determining regions (LCDRs) comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); (ii) D is a splicing modulator; (iii) L is a linker comprising ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15; and (iv) p is an integer from 1 to 15. It has.

[0398] In some embodiments, an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2 comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, an antibody or antigen-binding fragment thereof that targets neoplastic cells expressing EPHA2 comprises a human IgG1 heavy chain constant domain and a human Ig kappa light chain constant domain. In some embodiments, the antibody is 1C1. In some embodiments, p is an integer from 1 to 10, 2 to 8, or 4 to 8. In some embodiments, p is 4. In some embodiments, p is 8.

[0399] Drug portion The drug moiety (D) of the ADCs described herein can be any chemotherapeutic agent. Useful classes of chemotherapeutic agents include, for example, RNA splicing modulators. In certain preferred embodiments, the drug moiety is a splicing modulator. Exemplary splicing modulator compounds are described and exemplified herein.

[0400] In various embodiments, the drug moiety has the formula (II): [ka] (In the formula, R 1 is selected from absent, hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; and R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR 17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1 to C6 alkyl group, and -NR 15 R 16 Selected from; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 Selected from; R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; and Z is [ka] Selected from; R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 At least one of is hydrogen) or a pharmaceutically acceptable salt thereof.

[0401] In some embodiments, R 1 is selected from hydrogen, a C1-C4 alkyl group, a C1-C4 alkylcarboxylic acid group, and a C3-C8 cycloalkyl group. 1 is hydrogen. In some embodiments, R 1 is a C1-C4 alkyl group. In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R 1 is a C1-C4 alkyl carboxylic acid group. In some embodiments, R 1 is —CH2CH2CH2CO2H. In some embodiments, R 1 is a C3-C8 cycloalkyl group. In some embodiments, R 1 is cycloheptyl.

[0402] In some embodiments, R 3 is selected from hydrogen, a C1-C4 alkyl group, a C1-C4 alkylalkoxy group, a C1-C4 alkylcarboxylic acid group, and a C1-C4 alkylhydroxy group. 3 is selected from hydrogen and a C1-C4 alkyl carboxylic acid group. 3 is hydrogen. In some embodiments, R 3 is a C1-C4 alkyl carboxylic acid group. In some embodiments, R 3 is -CH2CH2CO2H.

[0403] In some embodiments, R 4 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, a —O—C(═O)—(C1-C4 alkyl) group, and a C1-C4 alkyl group. 4 is hydrogen. In some embodiments, R 4 is hydroxyl. In some embodiments, R 4 is an —O—(C1-C4 alkyl) group. In some embodiments, R 4 is —OCH. In some embodiments, R 4 is —OCH2CH3. In some embodiments, R 4 is an —OC(═O)—(C1-C4 alkyl) group. In some embodiments, R 4 is —OC(═O)—CH. In some embodiments, R 4 is —OC(═O)—CH2CH3. In some embodiments, R 4 is a C1-C4 alkyl group. In some embodiments, R 4 is methyl. In some embodiments, R 4 is ethyl.

[0404] In some embodiments, R 5 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, and a C1-C4 alkyl group. 5is hydrogen. In some embodiments, R 5 is hydroxyl. In some embodiments, R 5 is an —O—(C1-C4 alkyl) group. In some embodiments, R 5 is a C1 to C4 alkyl group.

[0405] In some embodiments, R 6 is hydrogen. In some embodiments, R 7 is hydrogen. In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 where R 17 is hydrogen and C 1 ~C 4 In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 where R 17 is hydrogen. In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen. In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen, where R 17 is hydrogen and C 1 ~C 4 In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen, where R 17 is hydrogen. In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16 In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16where R 15 is H, and R 16 is hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16 where R 15 is H, and R 16 is hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 Selected from R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, and a C3-C8 heterocyclyl group. 6 HA-OR 17 In some embodiments, R 6 is -OC(=O)-R 17 In some embodiments, R 6 is C1-C6 alkyl. In some embodiments, R 6 is C1-C4 alkyl. In some embodiments, R 6 is C alkyl. In some embodiments, R 6 Ha-NR 15 R 16 In some embodiments, R 7 HA-OR 17 In some embodiments, R 7 is -OC(=O)-R 17 In some embodiments, R 7 is C1-C6 alkyl. In some embodiments, R 7 is C1-C4 alkyl. In some embodiments, R 7 is C alkyl. In some embodiments, R 7 Ha-NR 15 R 16 is.

[0406] In some embodiments, R8 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, and a (C1-C4 alkyl). 8 is hydrogen. In some embodiments, R 8 is a hydroxyl group. In some embodiments, R 8 is an —O—(C1-C4 alkyl) group. In some embodiments, R 8 is an —O—(C1 alkyl) group.

[0407] In some embodiments, R 15 is hydrogen. In some embodiments, R 15 is R 17 In some embodiments, R 15 is -C(=O)-R 17 In some embodiments, R 15 is -C(=O)-OR 17 is.

[0408] In some embodiments, R 16 is hydrogen. In some embodiments, R 16 is R 17 In some embodiments, R 16 is -C(=O)-R 17 In some embodiments, R 16 is -C(=O)-OR 17 is.

[0409] In some embodiments, R 17 is selected from hydrogen, a C1-C4 alkyl group, a C3-C6 cycloalkyl group, and a C3-C8 heterocyclyl group. 17 is hydrogen. In some embodiments, R 17 is a C1-C4 alkyl group. In some embodiments, R 17 is a C alkyl group. In some embodiments, R 17 is a C3-C6 cycloalkyl group. In some embodiments, R 17 is a C cycloalkyl group. In some embodiments, R17 is a C4 cycloalkyl group. In some embodiments, R 17 is a C5 cycloalkyl group. In some embodiments, R 17 is a C cycloalkyl group. In some embodiments, R 17 is a C3-C8 heterocyclyl group. 17 is a C heterocyclyl group. In some embodiments, R 17 is a C4 heterocyclyl group. In some embodiments, R 17 is a C5 heterocyclyl group. In some embodiments, R 17 is a C6 heterocyclyl group. In some embodiments, R 17 is a C7 heterocyclyl group. In some embodiments, R 17 is a C8 heterocyclyl group.

[0410] In some embodiments, Z is [ka] In some embodiments, Z is [ka] In some embodiments, Z is [ka] In some embodiments, Z is [ka] In some embodiments, Z is [ka] is.

[0411] In some embodiments, the splicing modulator compound of formula (II) has formula (II-A): [ka] (Wherein Z' is [ka] Selected from; All other variables are as defined for formula (II). As shown in Figure 1, for example, the linker L in the ADC of formula (I) is linked to the linker L.

[0412] In various other embodiments, the drug moiety has formula (IV): [ka] (In the formula, R 1 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, an -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; and R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1 to C6 alkyl group, and -NR 15 R 16 Selected from; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 selected from; and R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 At least one of is hydrogen) or a pharmaceutically acceptable salt thereof.

[0413] In some embodiments, R 1 is selected from hydrogen, a C1-C4 alkyl group, a C1-C4 alkylcarboxylic acid group, and a C3-C8 cycloalkyl group. 1 is hydrogen. In some embodiments, R 1 is a C1-C4 alkyl group. In some embodiments, R 1is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R 1 is a C1-C4 alkyl carboxylic acid group. In some embodiments, R 1 is —CH2CH2CH2CO2H. In some embodiments, R 1 is a C3-C8 cycloalkyl group. In some embodiments, R 1 is cycloheptyl.

[0414] In some embodiments, R 3 is selected from hydrogen, a C1-C4 alkyl group, a C1-C4 alkylalkoxy group, a C1-C4 alkylcarboxylic acid group, and a C1-C4 alkylhydroxy group. 3 is selected from hydrogen and a C1-C4 alkyl carboxylic acid group. 3 is hydrogen. In some embodiments, R 3 is a C1-C4 alkyl carboxylic acid group. In some embodiments, R 3 is -CH2CH2CO2H.

[0415] In some embodiments, R 4 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, a —O—C(═O)—(C1-C4 alkyl) group, and a C1-C4 alkyl group. 4 is hydrogen. In some embodiments, R 4 is hydroxyl. In some embodiments, R 4 is an —O—(C1-C4 alkyl) group. In some embodiments, R 4 is —OCH. In some embodiments, R 4 is —OCH2CH3. In some embodiments, R 4 is an —OC(═O)—(C1-C4 alkyl) group. In some embodiments, R 4 is —OC(═O)—CH. In some embodiments, R 4is —OC(═O)—CH2CH3. In some embodiments, R 4 is a C1-C4 alkyl group. In some embodiments, R 4 is methyl. In some embodiments, R 4 is ethyl.

[0416] In some embodiments, R 5 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, and a C1-C4 alkyl group. 5 is hydrogen. In some embodiments, R 5 is hydroxyl. In some embodiments, R 5 is an —O—(C1-C4 alkyl) group. In some embodiments, R 5 is a C1 to C4 alkyl group.

[0417] In some embodiments, R 6 is hydrogen. In some embodiments, R 7 is hydrogen. In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 where R 17 is hydrogen and C 1 ~C 4 In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 where R 17 is hydrogen. In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen. In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen, where R 17 is hydrogen and C 1 ~C 4In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen, where R 17 is hydrogen. In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16 In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16 where R 15 is H, and R 16 is hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16 where R 15 is H, and R 16 is hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 Selected from R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, and a C3-C8 heterocyclyl group. 6 HA-OR 17 In some embodiments, R 6 is -OC(=O)-R 17 In some embodiments, R 6 is C1-C6 alkyl. In some embodiments, R 6 is C1-C4 alkyl. In some embodiments, R 6 is C alkyl. In some embodiments, R 6 Ha-NR 15 R 16 is.

[0418] In some embodiments, R 7HA-OR 17 In some embodiments, R 7 is -OC(=O)-R 17 In some embodiments, R 7 is C1-C6 alkyl. In some embodiments, R 7 is C1-C4 alkyl. In some embodiments, R 7 is C alkyl. In some embodiments, R 7 Ha-NR 15 R 16 is.

[0419] In some embodiments, R 8 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, and a (C1-C4 alkyl). 8 is hydrogen. In some embodiments, R 8 is a hydroxyl group. In some embodiments, R 8 is an —O—(C1-C4 alkyl) group. In some embodiments, R 8 is an —O—(C1 alkyl) group.

[0420] In some embodiments, R 15 is hydrogen. In some embodiments, R 15 is R 17 In some embodiments, R 15 is -C(=O)-R 17 In some embodiments, R 15 is -C(=O)-OR 17 is.

[0421] In some embodiments, R 16 is hydrogen. In some embodiments, R 16 is R 17 In some embodiments, R 16 is -C(=O)-R 17 In some embodiments, R 16 is -C(=O)-OR 17 is.

[0422] In some embodiments, R 17 is selected from hydrogen, a C1-C4 alkyl group, a C3-C6 cycloalkyl group, and a C3-C8 heterocyclyl group. 17 is hydrogen. In some embodiments, R 17 is a C1-C4 alkyl group. In some embodiments, R 17 is a C alkyl group. In some embodiments, R 17 is a C3-C6 cycloalkyl group. In some embodiments, R 17 is a C cycloalkyl group. In some embodiments, R 17 is a C4 cycloalkyl group. In some embodiments, R 17 is a C5 cycloalkyl group. In some embodiments, R 17 is a C cycloalkyl group. In some embodiments, R 17 is a C3-C8 heterocyclyl group. 17 is a C heterocyclyl group. In some embodiments, R 17 is a C4 heterocyclyl group. In some embodiments, R 17 is a C5 heterocyclyl group. In some embodiments, R 17 is a C6 heterocyclyl group. In some embodiments, R 17 is a C7 heterocyclyl group. In some embodiments, R 17 is a C8 heterocyclyl group.

[0423] In some embodiments, the splicing modulator compound of formula (IV) has formula (IV-A): [ka] As shown in Figure 1, for example, the linker L in the ADC of formula (I) is linked to the linker L.

[0424] In various other embodiments, the drug moiety has formula (VI): [ka] (In the formula, R 1 and R 9 are each independently selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; R 4 , R 5 , and R 8 are each independently selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —OC(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; R 6 and R 7 are each independently hydrogen, -OR 17 , -OC(=O)-R 17 , -OC(=O)-NR 15 R 16 , C1-C6 alkyl group, -NR 15 R 16 and a linker; R 10 is selected from hydrogen, a C1-C6 alkyl group, a -C(=O)-(C1-C6 alkyl) group, and -CD3; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 Selected from; R 17is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; and a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; R 6 and R 7 at least one of is hydrogen; R 1 and R 9 (It is impossible for both to exist) or a pharmaceutically acceptable salt thereof.

[0425] In some embodiments, R 1 is selected from hydrogen, a C1-C4 alkyl group, a C1-C4 alkylcarboxylic acid group, and a C3-C8 cycloalkyl group. 1 is hydrogen. In some embodiments, R 1 is a C1-C4 alkyl group. In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R 1 is a C1-C4 alkyl carboxylic acid group. In some embodiments, R 1 is —CH2CH2CH2CO2H. In some embodiments, R1 is a C3-C8 cycloalkyl group. In some embodiments, R 1 is cycloheptyl.

[0426] In some embodiments, R 3 is selected from hydrogen, a C1-C4 alkyl group, a C1-C4 alkylalkoxy group, a C1-C4 alkylcarboxylic acid group, and a C1-C4 alkylhydroxy group. 3 is selected from hydrogen and a C1-C4 alkyl carboxylic acid group. 3 is hydrogen. In some embodiments, R 3 is a C1-C4 alkyl carboxylic acid group. In some embodiments, R 3 is -CH2CH2CO2H.

[0427] In some embodiments, R 4 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, a —O—C(═O)—(C1-C4 alkyl) group, and a C1-C4 alkyl group. 4 is hydrogen. In some embodiments, R 4 is hydroxyl. In some embodiments, R 4 is an —O—(C1-C4 alkyl) group. In some embodiments, R 4 is —OCH. In some embodiments, R 4 is —OCH2CH3. In some embodiments, R 4 is an —OC(═O)—(C1-C4 alkyl) group. In some embodiments, R 4 is —OC(═O)—CH. In some embodiments, R 4 is —OC(═O)—CH2CH3. In some embodiments, R 4 is a C1-C4 alkyl group. In some embodiments, R 4 is methyl. In some embodiments, R 4 is ethyl.

[0428] In some embodiments, R 5 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, and a C1-C4 alkyl group. 5 is hydrogen. In some embodiments, R 5 is hydroxyl. In some embodiments, R 5 is an —O—(C1-C4 alkyl) group. In some embodiments, R 5 is a C1 to C4 alkyl group.

[0429] In some embodiments, R 9 is selected from absent, hydrogen, a C1-C4 alkyl group, —(C═O)—(C1-C4 alkyl) group, and —CD3. 9 is absent. In some embodiments, R 9 is hydrogen. In some embodiments, R 9 is a C1-C4 alkyl group. In some embodiments, the C1-C4 alkyl group is methyl. In some embodiments, the C1-C4 alkyl group is ethyl. In some embodiments, R 9 is a -(C=O)-(C1-C4 alkyl) group. In some embodiments, the -(C=O)-(C1-C4 alkyl) group is -(C=O)-methyl. In some embodiments, R 9 is -CD3.

[0430] In some embodiments, R 10 is selected from hydrogen, a C1-C4 alkyl group, a —(C═O)—(C1-C4 alkyl) group, and —CD3. 10 is hydrogen. In some embodiments, R 10 is a C1-C4 alkyl group. In some embodiments, the C1-C4 alkyl group is methyl. In some embodiments, the C1-C4 alkyl group is ethyl. In some embodiments, R 10is a -(C=O)-(C1-C4 alkyl) group. In some embodiments, the -(C=O)-(C1-C4 alkyl) group is -(C=O)-methyl. In some embodiments, R 10 is -CD3.

[0431] In some embodiments, R 6 is hydrogen. In some embodiments, R 7 is hydrogen. In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 where R 17 is hydrogen and C 1 ~C 4 In some embodiments, R 6 is hydrogen, and R 7 HA-OR 17 where R 17 is hydrogen. In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen. In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen, where R 17 is hydrogen and C 1 ~C 4 In some embodiments, R 6 HA-OR 17 and R 7 is hydrogen, where R 17 is hydrogen. In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16 In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16 where R 15is H, and R 16 is hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 In some embodiments, R 6 is hydrogen, and R 7 Ha-NR 15 R 16 where R 15 is H, and R 16 is hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 Selected from R 17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, and a C3-C8 heterocyclyl group. 6 HA-OR 17 In some embodiments, R 6 is -OC(=O)-R 17 In some embodiments, R 6 is C1-C6 alkyl. In some embodiments, R 6 is C1-C4 alkyl. In some embodiments, R 6 is C alkyl. In some embodiments, R 6 Ha-NR 15 R 16 is.

[0432] In some embodiments, R 7 HA-OR 17 In some embodiments, R 7 is -OC(=O)-R 17 In some embodiments, R 7 is C1-C6 alkyl. In some embodiments, R 7 is C1-C4 alkyl. In some embodiments, R 7 is C alkyl. In some embodiments, R 7 Ha-NR 15 R 16 is.

[0433] In some embodiments, R 8 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, and a (C1-C4 alkyl). 8 is hydrogen. In some embodiments, R 8 is a hydroxyl group. In some embodiments, R 8 is an —O—(C1-C4 alkyl) group. In some embodiments, R 8 is an —O—(C1 alkyl) group.

[0434] In some embodiments, R 15 is hydrogen. In some embodiments, R 15 is R 17 In some embodiments, R 15 is -C(=O)-R 17 In some embodiments, R 15 is -C(=O)-OR 17 is.

[0435] In some embodiments, R 16 is hydrogen. In some embodiments, R 16 is R 17 In some embodiments, R 16 is -C(=O)-R 17 In some embodiments, R 16 is -C(=O)-OR 17 is.

[0436] In some embodiments, R 17 is selected from hydrogen, a C1-C4 alkyl group, a C3-C6 cycloalkyl group, and a C3-C8 heterocyclyl group. 17 is hydrogen. In some embodiments, R 17 is a C1-C4 alkyl group. In some embodiments, R 17 is a C alkyl group. In some embodiments, R 17 is a C3-C6 cycloalkyl group. In some embodiments, R 17is a C cycloalkyl group. In some embodiments, R 17 is a C4 cycloalkyl group. In some embodiments, R 17 is a C5 cycloalkyl group. In some embodiments, R 17 is a C cycloalkyl group. In some embodiments, R 17 is a C3-C8 heterocyclyl group. 17 is a C heterocyclyl group. In some embodiments, R 17 is a C4 heterocyclyl group. In some embodiments, R 17 is a C5 heterocyclyl group. In some embodiments, R 17 is a C6 heterocyclyl group. In some embodiments, R 17 is a C7 heterocyclyl group. In some embodiments, R 17 is a C8 heterocyclyl group.

[0437] In some embodiments, a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, a is 1, 2, 3, 4, 5, or 6. In some embodiments, a is 1, 2, 3, 4, or 5. In some embodiments, a is 1, 2, 3, or 4. In some embodiments, a is 1, 2, or 3. In some embodiments, a is 1 or 2. In some embodiments, a is 1. In some embodiments, a is 2. In some embodiments, a is 3. In some embodiments, a is 4. In some embodiments, a is 5. In some embodiments, a is 6. In some embodiments, a is 7. In some embodiments, a is 8. In some embodiments, a is 9. In some embodiments, a is 10.

[0438] In some embodiments, the splicing modulator compound of formula (VI) has formula (VI-A): [ka] As shown in Figure 1, for example, the linker L in the ADC of formula (I) is linked to the linker L.

[0439] In various other embodiments, the drug moiety has formula (VIII): [ka] (In the formula, R 1 is selected from absent, hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, -OC(=O)-(C1-C6 alkyl) group, and -CD3; R 3 is selected from hydrogen, a C1-C6 alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylamino group, a C1-C6 alkylcarboxylic acid group, a C1-C6 alkylhydroxy group, a C3-C8 cycloalkyl group, a benzyl group, a C3-C8 heterocyclyl group, and an -OC(=O)-(C1-C6 alkyl) group; R 4 is selected from hydrogen, a hydroxyl group, a —O—(C1-C6 alkyl) group, a —O—C(═O)—(C1-C6 alkyl) group, and a C1-C6 alkyl group; and R 10 is selected from a 3- to 10-membered carbocyclic ring and a 3- to 10-membered heterocyclic ring, each of which is selected from 0 to 3 R a Each R is replaced by a are independently a halogen, a C1-C6 alkyl group, an -O-(C1-C6) alkyl group, a C1-C6 alkylalkoxy group, a C1-C6 alkylhydroxy group, or -S(=O) w -(4- to 7-membered heterocycle), a 4- to 7-membered carbocycle, and a 4- to 7-membered heterocycle; R 15 and R 16 are each independently hydrogen, R 17 , -C(=O)-R 17 , and -C(=O)-OR 17 selected from; and R17 is selected from hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, a benzyl group, and a C3-C8 heterocyclyl group; R 1 , R 3 , R 4 , R 10 , R 15 , R 16 , and R 17 are each independently a halogen, a hydroxyl group, a C1-C6 alkyl group, an -O-(C1-C6 alkyl) group, or -NR 15 R 16 , substituted by 0 to 3 groups independently selected from a C3 to C8 cycloalkyl group, a C1 to C6 alkylhydroxy group, a C1 to C6 alkylalkoxy group, a benzyl group, and a C3 to C8 heterocyclyl group; Each R a are independently a halogen, a hydroxyl group, or -NR 15 R 16 , C1-C6 alkyl group, -(C=O)-(C1-C6 alkyl) group, -(C=O)-(C1-C6 alkyl)-(C3-C 10 heterocyclyl groups), and C1-C6 alkylcarboxylic acid groups (wherein each of these is selected from the group consisting of halogen, hydroxyl, -NR 15 R 16 and C1-C3 alkyl; and w is 0, 1, or 2) or a pharmaceutically acceptable salt thereof.

[0440] In some embodiments, R 1 is selected from absent, hydrogen, a C1-C4 alkyl group, a C1-C4 alkylcarboxylic acid group, and a C3-C8 cycloalkyl group. 1 is hydrogen. In some embodiments, R 1 is a C1-C4 alkyl group. In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R1 is a C1-C4 alkyl carboxylic acid group. In some embodiments, R 1 is —CH2CH2CH2CO2H. In some embodiments, R 1 is a C3-C8 cycloalkyl group. In some embodiments, R 1 is cycloheptyl.

[0441] In some embodiments, R 3 is selected from hydrogen, a C1-C4 alkyl group, a C1-C4 alkylalkoxy group, a C1-C4 alkylcarboxylic acid group, and a C1-C4 alkylhydroxy group. 3 is selected from hydrogen and a C1-C4 alkyl carboxylic acid group. 3 is hydrogen. In some embodiments, R 3 is a C1-C4 alkyl carboxylic acid group. In some embodiments, R 3 is -CH2CH2CO2H.

[0442] In some embodiments, R 4 is selected from hydrogen, a hydroxyl group, a —O—(C1-C4 alkyl) group, a —O—C(═O)—(C1-C4 alkyl) group, and a C1-C4 alkyl group. 4 is hydrogen. In some embodiments, R 4 is hydroxyl. In some embodiments, R 4 is an —O—(C1-C4 alkyl) group. In some embodiments, R 4 is —OCH. In some embodiments, R 4 is —OCH2CH3. In some embodiments, R 4 is an —OC(═O)—(C1-C4 alkyl) group. In some embodiments, R 4 is —OC(═O)—CH. In some embodiments, R 4 is —OC(═O)—CH2CH3. In some embodiments, R 4 is a C1-C4 alkyl group. In some embodiments, R4 is methyl. In some embodiments, R 4 is ethyl.

[0443] In some embodiments, R 10 is selected from a 6- to 9-membered carbocyclic ring and a 6- to 9-membered heterocyclic ring, each of which is selected from 0 to 2 R a Each R is replaced by a is independently substituted with 0 to 3 groups independently selected from a halogen, a hydroxyl group, a C1-C6 alkyl group, a -(C=O)-(C1-C6 alkyl) group, a -(C=O)-(C1-C6 alkyl)-(3- to 10-membered heterocyclic) group, and a C1-C6 alkylcarboxylic acid group.

[0444] In some embodiments, the carbocycle may contain 0 to 2 R a wherein each R a is independently substituted with 0-3 groups independently selected from halogen, hydroxyl, C1-C6 alkyl, -(C=O)-(C1-C6 alkyl), -(C=O)-(C1-C6 alkyl)-(3-10 membered heterocycle), and C1-C6 alkylcarboxylic acid. a Each R is replaced by a is independently substituted with 0 to 3 groups independently selected from halogen, hydroxyl, C1-C6 alkyl, -(C=O)-(C1-C6 alkyl), -(C=O)-(C1-C6 alkyl)-(3-10 membered heterocycle), and C1-C6 alkylcarboxylic acid. [ka] is.

[0445] In some embodiments, the heterocycle may comprise 0 to 2 R a wherein each R ais independently substituted with 0 to 3 groups independently selected from halogen, hydroxyl, C1-C6 alkyl, -(C=O)-(C1-C6 alkyl), -(C=O)-(C1-C6 alkyl)-(3-10 membered heterocycle), and C1-C6 alkylcarboxylic acid. In some embodiments, the 9-membered heterocycle is [ka] is.

[0446] In some embodiments, R a is selected from halogen, a 3- to 10-membered carbocyclic ring, and a 3- to 10-membered heterocyclic ring, a is independently substituted with 0 to 3 groups independently selected from halogen, hydroxyl, C1-C6 alkyl, -(C=O)-(C1-C6 alkyl), -(C=O)-(C1-C6 alkyl)-(3-10 membered heterocyclic) and C1-C6 alkyl carboxylic acid groups. a is a halogen, [ka] is selected from.

[0447] In some embodiments, R 15 is hydrogen. In some embodiments, R 15 is R 17 In some embodiments, R 15 is -C(=O)-R 17 In some embodiments, R 15 is -C(=O)-OR 17 is.

[0448] In some embodiments, R 16 is hydrogen. In some embodiments, R 16 is R 17 In some embodiments, R 16 is -C(=O)-R 17 In some embodiments, R16 is -C(=O)-OR 17 is.

[0449] In some embodiments, R 17 is selected from hydrogen, a C1-C4 alkyl group, a C3-C6 cycloalkyl group, and a C3-C8 heterocyclyl group. 17 is hydrogen. In some embodiments, R 17 is a C1-C4 alkyl group. In some embodiments, R 17 is a C alkyl group. In some embodiments, R 17 is a C3-C6 cycloalkyl group. In some embodiments, R 17 is a C cycloalkyl group. In some embodiments, R 17 is a C4 cycloalkyl group. In some embodiments, R 17 is a C5 cycloalkyl group. In some embodiments, R 17 is a C cycloalkyl group. In some embodiments, R 17 is a C3-C8 heterocyclyl group. 17 is a C heterocyclyl group. In some embodiments, R 17 is a C4 heterocyclyl group. In some embodiments, R 17 is a C5 heterocyclyl group. In some embodiments, R 17 is a C6 heterocyclyl group. In some embodiments, R 17 is a C7 heterocyclyl group. In some embodiments, R 17 is a C8 heterocyclyl group.

[0450] In some embodiments, the splicing modulator compound of formula (VIII) has formula (VIII-A): [ka] As shown in Figure 1, for example, the linker L in the ADC of formula (I) is linked to the linker L.

[0451] In various embodiments, the drug moiety is a splicing modulator selected from D2 and D1.

[0452] In various embodiments, the drug moiety is D2. In various embodiments, the structure of the D2 drug moiety used in the disclosed ADCs is shown below: [ka]

[0453] In various embodiments, the linker in an ADC described herein (e.g., an ADC of Formula (I)) is covalently attached to a D2 drug moiety via the amine of a piperazine group. In various embodiments, the drug moiety is a derivative of D2. In various embodiments, a D2 derivative retains at least one biological function or activity of D2 (e.g., SF3b complex binding, in vitro splicing activity, cytotoxicity) but has an altered chemical structure.

[0454] In various embodiments, the drug moiety is D1 or a pharmaceutically acceptable salt thereof. In various embodiments, the structure of the D1 drug moiety used in the disclosed ADCs is shown below: [ka]

[0455] In various embodiments, the linker in an ADC described herein (e.g., an ADC of Formula (I)) is covalently attached to the D1 drug moiety via the amine of the piperazine group. In various embodiments, the drug moiety is a derivative of D1. In various embodiments, the D1 derivative retains at least one biological function or activity as D1 (e.g., SF3b complex binding, in vitro splicing activity, cytotoxicity) but has an altered chemical structure.

[0456] In some embodiments, the splicing modulator is D1: [ka] Includes.

[0457] In some embodiments, the splicing modulator is D2: [ka] Includes.

[0458] In some embodiments, the splicing modulator is D3: [ka] Includes.

[0459] In some embodiments, the splicing modulator is D4: [ka] Includes.

[0460] In some embodiments, the splicing modulator is D4': [ka] Includes.

[0461] In some embodiments, the splicing modulator is D5: [ka] Includes.

[0462] In some embodiments, the splicing modulator is D6: [ka] Includes.

[0463] In some embodiments, the splicing modulator is D7: [ka] Includes.

[0464] In some embodiments, the splicing modulator is D8: [ka] Includes.

[0465] In some embodiments, the splicing modulator is D9: [ka] Includes.

[0466] In some embodiments, the splicing modulator is D10: [ka] Includes.

[0467] In some embodiments, the splicing modulator is D11: [ka] Includes.

[0468] In some embodiments, the splicing modulator is D12: [ka] Includes.

[0469] In some embodiments, the splicing modulator is D13: [ka] Includes.

[0470] In some embodiments, the splicing modulator is D14: [ka] Includes.

[0471] In some embodiments, the splicing modulator is D15: [ka] Includes.

[0472] In some embodiments, the splicing modulator is D16: [ka] Includes.

[0473] In some embodiments, the splicing modulator is D17: [ka] Includes.

[0474] In some embodiments, the splicing modulator is D18: [ka] Includes.

[0475] In some embodiments, the splicing modulator is D19: [ka] Includes.

[0476] In some embodiments, the splicing modulator is D20: [ka] Includes.

[0477] In some embodiments, the splicing modulator is D21: [ka] Includes.

[0478] In some embodiments, the splicing modulator is D22: [ka] Includes.

[0479] In some embodiments, the splicing modulator is D23: [ka] Includes.

[0480] In some embodiments, the splicing modulator is D24: [ka] Includes.

[0481] In some embodiments, the splicing modulator is D25: [ka] Includes.

[0482] In some embodiments, the splicing modulator is D26: [ka] Includes.

[0483] In some embodiments, the splicing modulator is D27: [ka] Includes.

[0484] In some embodiments, the splicing modulator is D28: [ka] Includes.

[0485] In some embodiments, the splicing modulator is D29: [ka] Includes.

[0486] In some embodiments, the splicing modulator is D30: [ka] Includes.

[0487] In some embodiments, the splicing modulator is D31: [ka] Includes.

[0488] In some embodiments, the splicing modulator is D32: [ka] Includes.

[0489] In some embodiments, the splicing modulator is D33: [ka] Includes.

[0490] In some embodiments, the splicing modulator is D34: [ka] Includes.

[0491] In some embodiments, the splicing modulator is D35: [ka] Includes.

[0492] An exemplary ADC has formula (I): Ab-(LD) p (I) (In the formula, Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is D2; L is a linker covalently linking Ab to D; and p is an integer from 1 to 15. It has.

[0493] In some embodiments, the antibody or antigen-bind...

Claims

1. Formula (I): Ab-(L-D) ｐ (I) (In the formula, Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is a splicing modulator; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

2. 10. The antibody-drug conjugate of claim 1, wherein the antibody or antigen-binding fragment targets neoplastic cells derived from a hematological malignancy or a solid tumor.

3. 3. The antibody-drug conjugate of claim 1 or 2, wherein the antibody or antigen-binding fragment targets neoplastic cells derived from a hematological malignancy.

4. 4. The antibody-drug conjugate of claim 3, wherein the hematological malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

5. The antibody-drug conjugate of claim 3 or 4, wherein the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma.

6. 3. The antibody-drug conjugate of claim 1 or 2, wherein the antibody or antigen-binding fragment targets neoplastic cells derived from a solid tumor.

7. 7. The antibody-drug conjugate of claim 6, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, cervical cancer, pancreatic cancer, renal cancer, colorectal cancer, and esophageal cancer.

8. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets HER2-expressing cells.

9. The antibody-drug conjugate of claim 8, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

10. The antibody-drug conjugate of claim 8 or 9, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3).

11. The antibody-drug conjugate of any one of claims 8 to 10, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

12. The antibody-drug conjugate of any one of claims 8 to 11, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

13. The antibody-drug conjugate of any one of claims 8 to 12, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

14. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets CD138-expressing cells.

15. The antibody-drug conjugate of claim 14, wherein the antibody or antigen-binding fragment is an anti-CD138 antibody or antigen-binding fragment.

16. The antibody-drug conjugate of claim 14 or 15, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) 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 (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

17. The antibody-drug conjugate of any one of claims 14 to 16, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

22.

18. The antibody-drug conjugate of any one of claims 14 to 17, wherein the antibody or antigen-binding fragment comprises a human IgG2a heavy chain constant region.

19. The antibody-drug conjugate of any one of claims 14 to 18, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

20. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets an EPHA2-expressing cell.

21. The antibody-drug conjugate of claim 20, wherein the antibody or antigen-binding fragment is an anti-EPHA2 antibody or antigen-binding fragment.

22. The antibody-drug conjugate of claim 20 or 21, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3).

23. The antibody-drug conjugate of any one of claims 20 to 22, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

24.

24. The antibody-drug conjugate of any one of claims 20 to 23, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

25. The antibody-drug conjugate of any one of claims 20 to 24, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

26. The antibody-drug conjugate of any one of claims 1 to 25, wherein the linker is a cleavable linker.

27. The antibody-drug conjugate of claim 26 , wherein the linker comprises a cleavable peptide moiety.

28. 28. The antibody-drug conjugate of claim 27, wherein the cleavable peptide moiety is enzymatically cleavable.

29. The antibody-drug conjugate of any one of claims 26 to 28, wherein the cleavable peptide moiety or linker comprises an amino acid unit.

30. 30. The antibody-drug conjugate of claim 29, wherein the amino acid unit comprises valine-citrulline (Val-Cit).

31. 30. The antibody-drug conjugate of claim 29, wherein the amino acid unit comprises valine-alanine (Val-Ala).

32. The antibody-drug conjugate of claim 26, wherein the linker comprises a cleavable glucuronide moiety.

33. 33. The antibody-drug conjugate of claim 32, wherein the cleavable glucuronide moiety is enzymatically cleavable.

34. The antibody-drug conjugate of claim 32 or 33, wherein the cleavable glucuronide moiety is cleavable by a glucuronidase.

35. The antibody-drug conjugate of any one of claims 32 to 34, wherein the cleavable glucuronide moiety is cleavable by β-glucuronidase.

36. The antibody-drug conjugate of any one of claims 1 to 35, wherein the linker comprises at least one spacer unit.

37. 37. The antibody-drug conjugate of claim 36, wherein the spacer unit or linker comprises a polyethylene glycol (PEG) moiety.

38. The PEG moiety is -(PEG) ｍ and m is an integer from 1 to 10.

39. The antibody-drug conjugate of claim 38, wherein m is 2.

40. The antibody-drug conjugate of claim 36 , wherein the spacer unit or linker comprises an alkyl moiety.

41. The alkyl portion is —(CH ２ ) ｎ and n is an integer from 1 to 10.

42. The antibody-drug conjugate of claim 41, wherein n is 2.

43. The antibody-drug conjugate of claim 41, wherein n is 5.

44. The antibody-drug conjugate of claim 41, wherein n is 6.

45. 45. The antibody-drug conjugate of any one of claims 36 to 44, wherein the spacer unit is attached to the antibody or antigen-binding fragment via a maleimide (Mal) moiety ("Mal-spacer unit").

46. 46. ​​The antibody-drug conjugate of claim 45, wherein the Mal-spacer unit is reactive with a cysteine ​​residue on the antibody or antigen-binding fragment.

47. 47. The antibody-drug conjugate of claim 45 or 46, wherein the Mal-spacer unit is attached to the antibody or antigen-binding fragment via a cysteine ​​residue on the antibody or antigen-binding fragment.

48. The antibody-drug conjugate of any one of claims 45 to 47, wherein the linker comprises the Mal-spacer unit and a cleavable peptide moiety.

49. 49. The antibody-drug conjugate of claim 48, wherein the cleavable peptide moiety comprises an amino acid unit.

50. 50. The antibody-drug conjugate of claim 48 or 49, wherein the cleavable peptide moiety or amino acid unit comprises Val-Cit.

51. 50. The antibody-drug conjugate of claim 48 or 49, wherein the cleavable peptide moiety or amino acid unit comprises Val-Ala.

52. The antibody-drug conjugate of any one of claims 45 to 51, wherein the Mal-spacer unit comprises an alkyl moiety.

53. The antibody-drug conjugate of any one of claims 45 to 51, wherein the Mal-spacer unit comprises a PEG moiety.

54. The antibody-drug conjugate of any one of claims 45 to 52, wherein the Mal-spacer unit comprises maleimidocaproyl (MC).

55. 55. The antibody-drug conjugate of any one of claims 45 to 54, wherein the Mal-spacer unit links the antibody or antigen-binding fragment to the cleavable portion of the linker.

56. 56. The antibody-drug conjugate of claim 55, wherein the cleavable portion of the linker comprises a cleavable peptide moiety.

57. 57. The antibody-drug conjugate of claim 56, wherein the cleavable peptide moiety comprises an amino acid unit.

58. 58. The antibody-drug conjugate of claim 56 or 57, wherein the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn.

59. The antibody-drug conjugate of any one of claims 55 to 58, wherein the linker comprises MC-Val-Cit.

60. The antibody-drug conjugate of any one of claims 55 to 58, wherein the linker comprises MC-Val-Ala.

61. The antibody-drug conjugate of any one of claims 55 to 60, wherein the Mal-spacer unit comprises an alkyl moiety.

62. The antibody-drug conjugate of any one of claims 55 to 60, wherein the Mal-spacer unit comprises a PEG moiety.

63. 62. The antibody-drug conjugate of any one of claims 55 to 61, wherein the Mal-spacer unit comprises maleimidocaproyl (MC).

64. The antibody-drug conjugate of any one of claims 36 to 63, wherein the cleavable portion of the linker is directly joined to the splicing regulator or a spacer unit connects the cleavable portion of the linker to the splicing regulator.

65. The antibody-drug conjugate of claim 64, wherein cleavage of the conjugate releases the splicing modulator from the antibody or antigen-binding fragment and linker.

66. 66. The antibody-drug conjugate of claim 64 or 65, wherein the spacer unit connecting the cleavable portion of the linker to the splicing modulator is self-immolative.

67. 67. The antibody-drug conjugate of any one of claims 64 to 66, wherein the spacer unit connecting the cleavable portion of the linker to the splicing modulator comprises p-aminobenzyloxycarbonyl (pABC).

68. 68. The antibody-drug conjugate of claim 67, wherein the pABC links the cleavable portion of the linker to the splicing modulator.

69. 69. The antibody-drug conjugate of claim 67 or 68, wherein the cleavable portion of the linker comprises a cleavable peptide moiety.

70. 70. The antibody-drug conjugate of claim 69, wherein the cleavable peptide moiety comprises an amino acid unit.

71. 71. The antibody-drug conjugate of claim 69 or 70, wherein the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn.

72. The antibody-drug conjugate of any one of claims 67 to 71, wherein the linker comprises Val-Cit-pABC.

73. The antibody-drug conjugate of any one of claims 67 to 71, wherein the linker comprises Val-Ala-pABC.

74. 67. The antibody-drug conjugate of any one of claims 64 to 66, wherein the spacer unit connecting the cleavable portion of the linker to the splicing modulator comprises p-aminobenzyl (pAB).

75. 75. The antibody-drug conjugate of claim 74, wherein the pAB links the cleavable portion of the linker to the splicing modulator.

76. 76. The antibody-drug conjugate of claim 74 or 75, wherein the cleavable portion of the linker comprises a cleavable peptide moiety.

77. 77. The antibody-drug conjugate of claim 76, wherein the cleavable peptide moiety comprises an amino acid unit.

78. 78. The antibody-drug conjugate of claim 76 or 77, wherein the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn.

79. The antibody-drug conjugate of any one of claims 74 to 78, wherein the linker comprises Val-Cit-pAB.

80. The antibody-drug conjugate of any one of claims 74 to 78, wherein the linker comprises Val-Ala-pAB.

81. The antibody-drug conjugate of any one of claims 1 to 25, wherein the linker is a non-cleavable linker.

82. 82. The antibody-drug conjugate of claim 81, wherein the linker comprises at least one spacer unit.

83. 83. The antibody-drug conjugate of claim 81 or 82, wherein the spacer unit or linker comprises a polyethylene glycol (PEG) moiety.

84. The PEG moiety is -(PEG) ｍ - and m is an integer from 1 to 10.

85. The antibody-drug conjugate of claim 84, wherein m is 2.

86. 83. The antibody-drug conjugate of claim 81 or 82, wherein the spacer unit or linker comprises an alkyl moiety.

87. The alkyl portion is —(CH ２ ) ｎ - and n is an integer from 1 to 10.

88. 88. The antibody-drug conjugate of claim 87, wherein n is 2.

89. 88. The antibody-drug conjugate of claim 87, wherein n is 5.

90. 88. The antibody-drug conjugate of claim 87, wherein n is 6.

91. 91. The antibody-drug conjugate of any one of claims 81 to 90, wherein the spacer unit is attached to the antibody or antigen-binding fragment via a maleimide (Mal) moiety ("Mal-spacer unit").

92. 92. The antibody-drug conjugate of claim 91, wherein the Mal-spacer unit is reactive with a cysteine ​​residue on the antibody or antigen-binding fragment.

93. 93. The antibody-drug conjugate of claim 91 or 92, wherein the Mal-spacer unit is attached to the antibody or antigen-binding fragment via a cysteine ​​residue on the antibody or antigen-binding fragment.

94. The antibody-drug conjugate of any one of claims 91 to 93, wherein the Mal-spacer unit comprises an alkyl moiety.

95. The antibody-drug conjugate of any one of claims 91 to 93, wherein the Mal-spacer unit comprises a PEG moiety.

96. 96. The antibody-drug conjugate of any one of claims 91 to 95, wherein the linker or Mal-spacer unit comprises maleimidocaproyl (MC).

97. 97. The antibody-drug conjugate of claim 96, wherein the linker or Mal-spacer unit comprises maleimidocaproyl (MC) and at least one additional spacer unit.

98. The linker or Mal-spacer unit is MC-(PEG) ２ 96. The antibody-drug conjugate of any one of claims 91 to 95, comprising:

99. The linker or Mal-spacer unit is MC-(PEG) ２ and at least one additional spacer unit.

100. 96. The antibody-drug conjugate of any one of claims 91 to 95, wherein the linker or Mal-spacer unit comprises Mal-Hex.

101. 101. The antibody-drug conjugate of claim 100, wherein the linker or Mal-spacer unit comprises Mal-Hex and at least one additional spacer unit.

102. 96. The antibody-drug conjugate of any one of claims 91 to 95, wherein the linker or Mal-spacer unit comprises Mal-Et.

103. 103. The antibody-drug conjugate of claim 102, wherein the linker or Mal-spacer unit comprises Mal-Et and at least one additional spacer unit.

104. 96. The antibody-drug conjugate of any one of claims 91 to 95, wherein the linker or Mal-spacer unit comprises Mal-Et-O-Et.

105. 105. The antibody-drug conjugate of claim 104, wherein the linker or Mal-spacer unit comprises Mal-Et-O-Et and at least one additional spacer unit.

106. 106. The antibody-drug conjugate of any one of claims 91 to 105, wherein the Mal-spacer unit links the antibody or antigen-binding fragment to the splicing modulator.

107. The splicing regulator has the formula (II): 【Chemistry 1】 (In the formula, R １ Is absent, hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; and R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ Alkyl groups, and -NR １５ R １６ Selected from: R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ Selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; and Z is 【Chemistry 2】 Selected from: R １ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen) or a pharmaceutically acceptable salt thereof.

108. The linker is covalently attached to the splicing modulator ("LD"), wherein LD has the formula (II-A): 【Transformation 3】 (Wherein Z' is 【Chemistry 4】 Selected from: All other variables are as defined for formula (II).

108. The antibody-drug conjugate of claim 107, having the structure:

109. The splicing regulator has the formula (IV): 【Transformation 5】 (In the formula, R １ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; and R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ Alkyl groups, and -NR １５ R １６ Selected from: R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ is selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; R １ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen) The antibody-drug conjugate of any one of claims 1 to 106, which is a splicing regulator of the formula: or a pharmaceutically acceptable salt thereof.

110. The linker is covalently attached to the splicing modulator ("LD"), wherein LD has the formula (IV-A): 【Transformation 6】 110. The antibody-drug conjugate of claim 109, having the structure:

111. The splicing regulator has the formula (VI): 【Transformation 7】 (In the formula, R １ and R ９ are each independently hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ alkyl group, -NR １５ R １６ and a linker; R １０ is hydrogen, C １ ~C ６ Alkyl group, —C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ Selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; and a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; R １ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R ９ , R １０ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen; R １ and R ９ cannot both exist) The antibody-drug conjugate of any one of claims 1 to 106, which is a splicing regulator of the formula: or a pharmaceutically acceptable salt thereof.

112. The linker is covalently attached to the splicing modulator ("LD"), wherein LD has the formula (VI-A): 【Transformation 8】 112. The antibody-drug conjugate of claim 111, having the structure:

113. The splicing regulator has the formula (VIII): 【Chemistry 9】 (In the formula, R １ Is absent, hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; R ４ is hydrogen, a hydroxyl group, —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; and R １０ is selected from a 3- to 10-membered carbocycle and a 3- to 10-membered heterocycle, each of which is selected from 0 to 3 R ａ and each R ａ are independently halogen, C １ ~C ６ alkyl group, —O—(C １ ~C ６ ) alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylhydroxy group, —S(═O) ｗ -(4- to 7-membered heterocycle), 4- to 7-membered carbocycle, and 4- to 7-membered heterocycle; R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ is selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; R １ , R ３ , R ４ , R １０ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; Each R ａ are independently a halogen, a hydroxyl group, or —NR １５ R １６ , C １ ~C ６ Alkyl group, -(C=O)-(C １ ~C ６ alkyl) group, -(C=O)-(C １ ~C ６ alkyl)-(C ３ ~C １０ heterocyclyl group, —S(═O) ｗ -(C ３ ~C ８ heterocyclyl groups), and C １ ~C ６ alkylcarboxylic acid groups (wherein each of these is selected from the group consisting of halogen, hydroxyl, -NR １５ R １６ , and C １ ~C ３ substituted with 0 to 3 groups independently selected from alkyl; w is 0, 1, or 2. The antibody-drug conjugate of any one of claims 1 to 106, which is a splicing regulator of the formula: or a pharmaceutically acceptable salt thereof.

114. The linker is covalently attached to the splicing modulator ("LD"), wherein LD has the formula (VIII-A): 【Chemistry 10】 or a pharmaceutically acceptable salt thereof.

115. The antibody-drug conjugate of any one of claims 1 to 106, wherein the splicing modulator comprises a modulator of the SF3b complex.

116. The antibody-drug conjugate of claim 115, wherein the splicing modulator comprises a pladienolide or a pladienolide derivative.

117. The antibody-drug conjugate of claim 115 or 116, wherein the splicing modulator comprises pladienolide D or a pladienolide D derivative.

118. The antibody-drug conjugate of claim 117, wherein the pladienolide D or derivative comprises D2, D1, D4, D8, D10, D11 (E7107), D20, D21, D22, D12, or D25.

119. The antibody-drug conjugate of claim 117 or 118, wherein the pladienolide D or derivative comprises D2.

120. The antibody-drug conjugate of claim 117 or 118, wherein the pladienolide D or derivative comprises D1.

121. The antibody-drug conjugate of claim 117 or 118, wherein the pladienolide D or derivative is a zwitterionic pladienolide D or derivative.

122. The antibody-drug conjugate of claim 121, wherein the zwitterionic pladienolide D or derivative comprises D22 or D25.

123. The antibody-drug conjugate of claim 115 or 116, wherein the splicing modulator comprises pladienolide B or a pladienolide B derivative.

124. The antibody-drug conjugate of claim 123, wherein the pladienolide B or derivative comprises D9, D18, D19, or D13.

125. The antibody-drug conjugate of claim 115 or 116, wherein the splicing modulator comprises an arylpladienolide.

126. The antibody-drug conjugate of claim 125, wherein the arylpladienolide comprises D15, D14, D16, D17, D26, or D33.

127. The antibody-drug conjugate of claim 125 or 126, wherein the arylpladienolide is a zwitterionic arylpladienolide.

128. The antibody-drug conjugate of claim 127, wherein the zwitterionic arylpladienolide comprises D33.

129. The splicing regulator is D2: 【Chemistry 11】 The antibody-drug conjugate of any one of claims 1 to 106, comprising:

130. The splicing regulator is D1: 【Chemistry 12】 The antibody-drug conjugate of any one of claims 1 to 106, comprising:

131. The antibody-drug conjugate of any one of claims 1 to 106, wherein the splicing modulator comprises D4, D12, D15, D8, D9, D10, D13, D18, D19, D20, D21, D22, D25, or D33.

132. The antibody-drug conjugate of any one of claims 1 to 131, wherein p is 1 to 10.

133. The antibody-drug conjugate of any one of claims 1 to 132, wherein p is 2 to 8.

134. The antibody-drug conjugate of any one of claims 1 to 133, wherein p is 4 to 8.

135. The antibody-drug conjugate of any one of claims 1 to 134, wherein p is 4.

136. The antibody-drug conjugate of any one of claims 1 to 134, wherein p is 8.

137. Formula (I): Ab-(L-D) ｐ (I) (In the formula, Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is D2; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

138. The antibody-drug conjugate of claim 137, wherein the antibody or antigen-binding fragment targets a HER2-expressing cell.

139. The antibody-drug conjugate of claim 138, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

140. The antibody-drug conjugate of claim 138 or 139, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3).

141. The antibody-drug conjugate of any one of claims 138 to 140, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

142. The antibody-drug conjugate of any one of claims 138 to 141, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

143. The antibody-drug conjugate of any one of claims 138 to 142, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

144. The antibody-drug conjugate of claim 137, wherein the antibody or antigen-binding fragment targets CD138-expressing cells.

145. The antibody-drug conjugate of claim 144, wherein the antibody or antigen-binding fragment is an anti-CD138 antibody or antigen-binding fragment.

146. The antibody-drug conjugate of claim 144 or 145, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) 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 (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

147. The antibody-drug conjugate of any one of claims 144 to 146, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

22.

148. The antibody-drug conjugate of any one of claims 144 to 147, wherein the antibody or antigen-binding fragment comprises a human IgG2a heavy chain constant region.

149. The antibody-drug conjugate of any one of claims 144 to 148, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

150. The antibody-drug conjugate of claim 137, wherein the antibody or antigen-binding fragment targets an EPHA2-expressing cell.

151. The antibody-drug conjugate of claim 150, wherein the antibody or antigen-binding fragment is an anti-EPHA2 antibody or antigen-binding fragment.

152. The antibody-drug conjugate of claim 150 or 151, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3).

153. The antibody-drug conjugate of any one of claims 150 to 152, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

24.

154. The antibody-drug conjugate of any one of claims 150 to 153, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

155. The antibody-drug conjugate of any one of claims 150 to 154, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

156. Formula (I): Ab-(L-D) ｐ (I) (In the formula, Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is D1; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

157. The antibody-drug conjugate of claim 156, wherein the antibody or antigen-binding fragment targets a HER2-expressing cell.

158. The antibody-drug conjugate of claim 157, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

159. The antibody-drug conjugate of claim 157 or 158, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3).

160. The antibody-drug conjugate of any one of claims 157 to 159, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

161. The antibody-drug conjugate of any one of claims 157 to 160, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

162. The antibody-drug conjugate of any one of claims 157 to 161, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

163. The antibody-drug conjugate of claim 156, wherein the antibody or antigen-binding fragment targets CD138-expressing cells.

164. The antibody-drug conjugate of claim 163, wherein the antibody or antigen-binding fragment is an anti-CD138 antibody or antigen-binding fragment.

165. The antibody-drug conjugate of claim 163 or 164, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) 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 (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

166. The antibody-drug conjugate of any one of claims 163 to 165, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

22.

167. The antibody-drug conjugate of any one of claims 163 to 166, wherein the antibody or antigen-binding fragment comprises a human IgG2a heavy chain constant region.

168. The antibody-drug conjugate of any one of claims 163 to 167, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

169. The antibody-drug conjugate of claim 156, wherein the antibody or antigen-binding fragment targets an EPHA2-expressing cell.

170. The antibody-drug conjugate of claim 169, wherein the antibody or antigen-binding fragment is an anti-EPHA2 antibody or antigen-binding fragment.

171. The antibody-drug conjugate of claim 169 or 170, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3).

172. The antibody-drug conjugate of any one of claims 169 to 171, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

24.

173. The antibody-drug conjugate of any one of claims 169 to 172, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

174. The antibody-drug conjugate of any one of claims 169 to 173, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

175. Formula (I): Ab-(L-D) ｐ (I) (In the formula, The Ab is an anti-HER2 antibody or antigen-binding fragment comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO:1 (HCDR1), SEQ ID NO:2 (HCDR2), and SEQ ID NO:3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3); D is D2, D1, D4, D12, or D15; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

176. The antibody-drug conjugate of claim 175, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

177. Formula (I): Ab-(L-D) ｐ (I) (In the formula, The Ab is an anti-CD138 antibody or antigen-binding fragment comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) 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 (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3); D is D2, D1, D4, D12, or D15; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

178. The antibody-drug conjugate of claim 177, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

22.

179. Formula (I): Ab-(L-D) ｐ (I) (In the formula, The Ab is an anti-EPHA2 antibody or antigen-binding fragment comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3); D is D2, D1, D4, D12, or D15; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

180. The antibody-drug conjugate of claim 179, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

24.

181. The antibody-drug conjugate of any one of claims 156 to 180, wherein the linker is a cleavable linker.

182. The antibody-drug conjugate of claim 181, wherein the cleavable linker comprises MC-Val-Cit-pABC.

183. The antibody-drug conjugate of claim 181, wherein the cleavable linker comprises MC-Val-Ala-pABC.

184. The antibody-drug conjugate of claim 181, wherein the cleavable linker comprises MC-Val-Ala-pAB.

185. The antibody-drug conjugate of any one of claims 156 to 180, wherein the linker is a non-cleavable linker.

186. The antibody-drug conjugate of claim 185, wherein the non-cleavable linker comprises Mal-Hex.

187. The antibody-drug conjugate of claim 185, wherein the non-cleavable linker comprises Mal-Et.

188. The antibody-drug conjugate of claim 185, wherein the non-cleavable linker comprises Mal-Et-O-Et.

189. The antibody-drug conjugate of any one of claims 156 to 188, wherein p is 1 to 10.

190. The antibody-drug conjugate of any one of claims 156 to 189, wherein p is 2 to 8.

191. The antibody-drug conjugate of any one of claims 156 to 190, wherein p is 4 to 8.

192. The antibody-drug conjugate of any one of claims 156 to 191, wherein p is 4.

193. The antibody-drug conjugate of any one of claims 156 to 191, wherein p is 8.

194. 200. A composition comprising multiple copies of the antibody-drug conjugate of any one of claims 1 to 193, wherein the average p of said antibody-drug conjugates in said composition is from about 3.5 to about 5.

5.

195. The composition of claim 194, wherein the average p of the antibody-drug conjugates in the composition is about 4.

196. 200. A composition comprising multiple copies of the antibody-drug conjugate of any one of claims 1 to 193, wherein the average p of said antibody-drug conjugates in said composition is from about 7 to about 9.

197. The composition of claim 196, wherein the average p of the antibody-drug conjugates in the composition is about 8.

198. 197. A method of treating a subject having or suspected of having a neoplastic disorder, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 193 or the composition of any one of claims 194 to 197.

199. 199. The method of claim 198, wherein the neoplastic disorder is a hematological malignancy or a solid tumor.

200. 200. The method of claim 198 or 199, wherein the neoplastic disorder is a hematological malignancy.

201. 201. The method of claim 199 or 200, wherein the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma.

202. 200. The method of claim 198 or 199, wherein the neoplastic disorder is a solid tumor.

203. 202. The method of claim 199 or 202, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, cervical cancer, pancreatic cancer, renal cancer, colorectal cancer, and esophageal cancer.

204. 204. The method of any one of claims 198-203, wherein treatment with the antibody-drug conjugate or composition induces bystander killing of neoplastic cells that do not express the target antigen but are adjacent to neoplastic cells that do express the target antigen.

205. 205. The method of any one of claims 198-204, wherein the subject has one or more neoplastic cells that express a target antigen.

206. 206. The method of claim 205, wherein the target antigen is HER2.

207. 207. The method of claim 206, wherein the one or more neoplastic cells are derived from HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary duct cancer.

208. 208. The method of claim 207, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

209. The method of any one of claims 206 to 208, wherein the subject is refractory or poorly responsive to treatment with (a) an anti-HER2 antibody when administered alone and / or (b) a splicing modulator when administered alone.

210. The method of any one of claims 206 to 209, wherein the subject is intolerant, refractory, or poorly responsive to treatment with a splicing modulator when administered alone.

211. The method of claim 205, wherein the target antigen is CD138.

212. 212. The method of claim 211, wherein the one or more neoplastic cells are derived from multiple myeloma that expresses CD138.

213. The method of claim 211 or 212, wherein the subject is refractory or poorly responsive to treatment with (a) an anti-CD138 antibody when administered alone and / or (b) a splicing modulator when administered alone.

214. The method of any one of claims 211 to 213, wherein the subject is intolerant, refractory, or poorly responsive to treatment with a splicing modulator when administered alone.

215. 206. The method of claim 205, wherein the target antigen is EPHA2.

216. 216. The method of claim 215, wherein the one or more neoplastic cells are derived from an EPHA2-expressing breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer.

217. The method of claim 215 or 216, wherein the subject is refractory or poorly responsive to treatment with (a) an anti-EPHA2 antibody when administered alone and / or (b) a splicing modulator when administered alone.

218. The method of any one of claims 215 to 217, wherein the subject is intolerant, refractory, or poorly responsive to treatment with a splicing modulator when administered alone.

219. 197. A method of reducing or inhibiting tumor growth in a subject having or suspected of having a neoplastic disorder, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 1 to 193 or the composition of any one of claims 194 to 197.

220. 220. The method of claim 219, wherein treatment with the antibody-drug conjugate or composition induces bystander killing of neoplastic tumor cells that do not express the target antigen but are adjacent to neoplastic tumor cells that do express the target antigen.

221. 221. The method of claim 219 or 220, wherein the tumor comprises one or more neoplastic cells that express a target antigen.

222. 222. The method of claim 221, wherein the target antigen is HER2.

223. 223. The method of claim 222, wherein the one or more neoplastic cells are derived from HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary duct cancer.

224. 224. The method of claim 223, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

225. 225. The method of any one of claims 222 to 224, wherein the tumor is resistant or refractory to treatment with (a) an anti-HER2 antibody when administered alone and / or (b) a splicing modulator when administered alone.

226. The method of claim 221, wherein the target antigen is CD138.

227. The method of claim 226, wherein the one or more neoplastic cells are derived from a multiple myeloma that expresses CD138.

228. The method of claim 226 or 227, wherein the tumor is resistant or refractory to treatment with (a) an anti-CD138 antibody when administered alone and / or (b) a splicing modulator when administered alone.

229. 222. The method of claim 221, wherein the target antigen is EPHA2.

230. 230. The method of claim 229, wherein the one or more neoplastic cells are derived from an EPHA2-expressing breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer.

231. The method of claim 229 or 230, wherein the tumor is resistant or refractory to treatment with (a) an anti-EPHA2 antibody when administered alone and / or (b) a splicing modulator when administered alone.

232. 197. A method of determining whether a subject having or suspected of having a neoplastic disorder will respond to treatment with the antibody-drug conjugate of any one of claims 1 to 193 or the composition of any one of claims 194 to 197, the method comprising providing a biological sample from the subject and contacting the biological sample with the antibody-drug conjugate of any one of claims 1 to 193 or the composition of any one of claims 194 to 197.

233. 233. The method of claim 232, wherein the biological sample is a tumor sample.

234. 234. The method of claim 233, wherein the tumor sample is a tumor biopsy or a blood sample.

235. 235. The method of claim 234, wherein the blood sample is selected from blood, a blood fraction, or cells obtained from the blood or blood fraction.

236. 236. The method of any one of claims 232-235, wherein the subject has one or more neoplastic cells that express a target antigen.

237. 237. The method of claim 236, wherein the target antigen is HER2.

238. 238. The method of claim 237, wherein the one or more neoplastic cells are derived from HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary duct cancer.

239. 239. The method of claim 238, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

240. The method of claim 236, wherein the target antigen is CD138.

241. 241. The method of claim 240, wherein the one or more neoplastic cells are derived from multiple myeloma that expresses CD138.

242. 237. The method of claim 236, wherein the target antigen is EPHA2.

243. 243. The method of claim 242, wherein the one or more neoplastic cells are derived from an EPHA2-expressing breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer.

244. 200. An antibody-drug conjugate according to any one of claims 1 to 193 or a composition according to any one of claims 194 to 197 for use in the treatment of a neoplastic disorder.

245. 245. The antibody-drug conjugate for use according to claim 244, wherein the neoplastic disorder is characterized by having one or more neoplastic cells that express the target antigen.

246. The antibody-drug conjugate for use according to claim 245, wherein the target antigen is HER2.

247. 247. The antibody-drug conjugate for use according to claim 246, wherein the neoplastic disorder is HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary duct cancer.

248. The antibody-drug conjugate of claim 247, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

249. The antibody-drug conjugate for use according to claim 245, wherein the target antigen is CD138.

250. 250. The antibody-drug conjugate for use according to claim 249, wherein said neoplastic disorder is CD138-expressing multiple myeloma.

251. The antibody-drug conjugate for use according to claim 245, wherein the target antigen is EPHA2.

252. 252. The antibody-drug conjugate for use according to claim 251, wherein said neoplastic disorder is EPHA2-expressing breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer.

253. 202. Use of an antibody-drug conjugate according to any one of claims 1 to 193 or a composition according to any one of claims 194 to 197 in the treatment of a neoplastic disorder.

254. 254. The use of claim 253, wherein the neoplastic disorder is characterized by having one or more neoplastic cells that express the target antigen.

255. The use of claim 254, wherein the target antigen is HER2.

256. 256. The use of claim 255, wherein the neoplastic disorder is HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary duct cancer.

257. The use of claim 256, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

258. The use described in claim 254, wherein the target antigen is CD138.

259. The use of claim 258, wherein the neoplastic disorder is CD138-expressing multiple myeloma.

260. The use of claim 254, wherein the target antigen is EPHA2.

261. 261. The use of claim 260, wherein the neoplastic disorder is EPHA2-expressing breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer.

262. 202. Use of an antibody-drug conjugate according to any one of claims 1 to 193 or a composition according to any one of claims 194 to 197 in a method for the manufacture of a medicament for the treatment of a neoplastic disorder.

263. 263. The use of claim 262, wherein the neoplastic disorder is characterized by having one or more neoplastic cells that express the target antigen.

264. The use of claim 263, wherein the target antigen is HER2.

265. 265. The use of claim 264, wherein the neoplastic disorder is HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary duct cancer.

266. The use of claim 265, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

267. The use described in claim 263, wherein the target antigen is CD138.

268. The use of claim 267, wherein the neoplastic disorder is CD138-expressing multiple myeloma.

269. The use of claim 263, wherein the target antigen is EPHA2.

270. 270. The use of claim 269, wherein the neoplastic disorder is EPHA2-expressing breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer.

271. 198. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 1 to 193 or the composition of any one of claims 194 to 197, and a pharmaceutically acceptable carrier.

272. 200. A method of making an antibody-drug conjugate of any one of claims 1 to 193, comprising reacting an antibody or antigen-binding fragment with a linker tethered to a splicing modulator under conditions that allow conjugation.

273. Formula (III): 【Chemistry 13】 (In the formula, R １ Is absent, hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ２ is absent or a linker; R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; and R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ alkyl group, -NR １５ R １６ and a linker; R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ Selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; Z” is, 【Chemistry 14】 Selected from: R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen; R ２ is a linker, R ６ or R ７ is a linker, and R ６ or R ７ is a linker, R ２ does not exist) or a pharmaceutically acceptable salt thereof.

274. Formula (V): 【Chemistry 15】 (In the formula, R １ Is absent, hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ２ is absent or a linker; R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; and R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ alkyl group, -NR １５ R １６ and a linker; R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ is selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen; R ２ is a linker, R ６ or R ７ is a linker, and R ６ or R ７ is a linker, R ２ does not exist) or a pharmaceutically acceptable salt thereof.

275. Formula (VII): 【Chemistry 16】 (In the formula, R １ and R ９ are each independently absent, hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ２ is absent or a linker; R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ alkyl group, -NR １５ R １６ and a linker; R １０ is hydrogen, C １ ~C ６ Alkyl group, —C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ Selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; and a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R ９ , R １０ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen; R ２ is a linker, R ６ or R ７ is a linker, and R ６ or R ７ is a linker, R ２ does not exist; R １ and R ９ cannot both exist) or a pharmaceutically acceptable salt thereof.

276. Formula (IX): 【Chemistry 17】 (In the formula, R １ Is absent, hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ２ is a linker; R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; R ４ is hydrogen, a hydroxyl group, —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R １０ is selected from a 3- to 10-membered carbocycle and a 3- to 10-membered heterocycle, each of which is selected from 0 to 3 R ａ and each R ａ are independently halogen, C １ ~C ６ alkyl group, —O—(C １ ~C ６ ) alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylhydroxy group, —S(═O) ｗ -(4- to 7-membered heterocycle), 4- to 7-membered carbocycle, and 4- to 7-membered heterocycle; R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ is selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; R １ , R ２ , R ３ , R ４ , R １０ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; Each R ａ are independently a halogen, a hydroxyl group, or —NR １５ R １６ , C １ ~C ６ Alkyl group, -(C=O)-(C １ ~C ６ alkyl) group, -(C=O)-(C １ ~C ６ alkyl)-(C ３ ~C １０ heterocyclyl group, —S(═O) ｗ -(C ３ ~C ８ Heterocyclyl groups, and C １ ~C ６ alkylcarboxylic acid groups (wherein each of these is selected from the group consisting of halogen, hydroxyl, -NR １５ R １６ , and C １ ~C ３ substituted with 0 to 3 groups independently selected from alkyl groups; or a pharmaceutically acceptable salt thereof.

277. formula [Chemistry 18] and pharmaceutically acceptable salts thereof.

278. 278. A pharmaceutical composition comprising the compound of claim 277 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

279. formula 【Chemistry 19】 and pharmaceutically acceptable salts thereof.

280. 280. A pharmaceutical composition comprising the compound of claim 279 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

281. formula 【Chemistry 20】 and pharmaceutically acceptable salts thereof.

282. 282. A pharmaceutical composition comprising the compound of claim 281 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

283. formula 【Chemistry 21】 and pharmaceutically acceptable salts thereof.

284. 284. A pharmaceutical composition comprising the compound of claim 283 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

285. formula 【Chemistry 22】 and pharmaceutically acceptable salts thereof.

286. 286. A pharmaceutical composition comprising the compound of claim 285 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

287. formula 【Chemistry 23】 and pharmaceutically acceptable salts thereof.

288. 288. A pharmaceutical composition comprising the compound of claim 287 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

289. formula 【Chemistry 24】 and pharmaceutically acceptable salts thereof.

290. 290. A pharmaceutical composition comprising the compound of claim 289 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

291. formula 【Chemistry 25】 and pharmaceutically acceptable salts thereof.

292. 292. A pharmaceutical composition comprising the compound of claim 291 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

293. formula 【Chemistry 26】 and pharmaceutically acceptable salts thereof.

294. 294. A pharmaceutical composition comprising the compound of claim 293 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

295. formula 【Chemistry 27】 and pharmaceutically acceptable salts thereof.

296. 296. A pharmaceutical composition comprising the compound of claim 295 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

297. formula 【Chemistry 28】 and pharmaceutically acceptable salts thereof.

298. 300. A pharmaceutical composition comprising the compound of claim 297 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

299. formula 【Chemistry 29】 and pharmaceutically acceptable salts thereof.

300. 300. A pharmaceutical composition comprising the compound of claim 299 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

301. 300. A method of treating a subject having or suspected of having a neoplastic disorder, comprising administering to said subject a therapeutically effective amount of a compound or pharmaceutical composition according to any one of claims 273-300.

302. 300. A method of reducing or inhibiting tumor growth in a subject having or suspected of having a neoplastic disorder, comprising administering to said subject a therapeutically effective amount of a compound or pharmaceutical composition according to any one of claims 273-300.

303. 20. A method for inducing at least one neoantigen, comprising contacting neoplastic cells with an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1-193, or a composition of any one of claims 194-197, thereby inducing the production of at least one neoantigen.

304. 304. The method of claim 303, wherein said at least one neo-antigen comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

305. 305. The method of claim 304, wherein said at least one neoantigen comprises the amino acid sequence of SEQ ID NO:

37.

306. 305. The method of claim 304, wherein said at least one neoantigen comprises the amino acid sequence of SEQ ID NO:

39.

307. 305. The method of claim 304, wherein said at least one neo-antigen comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

308. 308. The method of any one of claims 303 to 307, wherein the neoplastic cells are in in vitro cell culture.

309. The method of claim 308, wherein the neoplastic cells are obtained from a subject.

310. The method of any one of claims 303 to 307, wherein the neoplastic cell is present in a subject.

311. 311. The method of any one of claims 303 to 310, wherein the neoplastic cells are derived from a hematological malignancy or a solid tumor.

312. 312. The method of claim 311, wherein the hematological malignancy is selected from a B-cell malignancy, leukemia, lymphoma, and myeloma.

313. 313. The method of claim 311 or 312, wherein the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma.

314. 312. The method of claim 311, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, cervical cancer, pancreatic cancer, renal cancer, colorectal cancer, and esophageal cancer.

315. 197. A method of inducing at least one neoantigen and / or T cell response in a subject having or suspected of having a neoplastic disorder, the method comprising administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1 to 193, or a composition of any one of claims 194 to 197.

316. 20. A method of treating a subject having or suspected of having a neoplastic disorder, comprising administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1 to 193, or a composition of any one of claims 194 to 197, wherein administration of the splicing modulator, antibody-drug conjugate, or composition induces at least one neo-antigen and / or T cell response.

317. 317. The method of claim 315 or 316, wherein said at least one neo-antigen comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

318. The method of claim 317, wherein said at least one neoantigen comprises the amino acid sequence of SEQ ID NO:

37.

319. The method of claim 317, wherein the at least one neoantigen comprises the amino acid sequence of SEQ ID NO:

39.

320. 318. The method of claim 317, wherein said at least one neo-antigen comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

321. The method of any one of claims 315 to 320, wherein the dosage of the splicing modulator, antibody-drug conjugate, or composition is reduced due to induction of at least one neo-antigen and / or T cell response when compared to a standard dosage of the splicing modulator, antibody-drug conjugate, or composition.

322. The method of any one of claims 315 to 321, wherein the dosage of the splicing modulator, antibody-drug conjugate, or composition is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to a standard dosage of the splicing modulator, antibody-drug conjugate, or composition.

323. The method of any one of claims 315 to 322, wherein the splicing modulator, antibody-drug conjugate, or composition is administered at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% less frequently when compared to a standard administration regimen of the splicing modulator, antibody-drug conjugate, or composition.

324. The method of any one of claims 315 to 323, wherein the administration amount and / or dosage of the splicing modulator, antibody-drug conjugate, or composition results in reduced systemic toxicity and / or improved tolerability.

325. 325. The method of any one of claims 315-324, further comprising administering at least one additional therapy.

326. The method of claim 325, wherein the dosage of the splicing modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy is reduced due to induction of at least one neo-antigen and / or T cell response when compared to standard dosages of the splicing modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy.

327. The method of claim 325 or 326, wherein the dosage of the splicing modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to standard dosages of the splicing modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy.

328. The method of any one of claims 325 to 327, wherein the splicing modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy is administered at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% less frequently when compared to a standard administration regimen of the splicing modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy.

329. The method of any one of claims 325 to 328, wherein the administration amount and / or dosage of the splicing modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy results in reduced systemic toxicity and / or improved tolerability.

330. 330. The method of any one of claims 325-329, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is initiated prior to administration of the at least one additional therapy.

331. 330. The method of any one of claims 325-329, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is initiated after administration of the at least one additional therapy.

332. 330. The method of any one of claims 325-329, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is initiated simultaneously with administration of the at least one additional therapy.

333. 333. The method of any one of claims 315 to 332, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is repeated at least once after an initial administration.

334. The method of claim 333, wherein the amount used in repeat administrations of the splicing modulator, antibody-drug conjugate, or composition is reduced when compared to the amount used in the initial administration.

335. The method of claim 333 or 334, wherein the amount used for repeated administration of the splicing modulator, antibody-drug conjugate, or composition is reduced when compared to a standard dosage of the splicing modulator, antibody-drug conjugate, or composition.

336. The method of any one of claims 333 to 335, wherein the amount used for repeat administration of the splicing modulator, antibody-drug conjugate, or composition is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to a standard dosage of the splicing modulator, antibody-drug conjugate, or composition.

337. 337. The method of any one of claims 325-336, wherein the administration of the at least one additional therapy is repeated at least once after an initial administration.

338. The method of claim 337, wherein the amount used in repeat administrations of the at least one additional therapy is reduced when compared to the amount used in the initial administration.

339. 339. The method of claim 337 or 338, wherein the amount used for repeat administration of the at least one additional therapy is reduced when compared to the standard dosage of the at least one additional therapy.

340. 340. The method of any one of claims 337-339, wherein the amount used for repeat administration of the at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to a standard dosage of the at least one additional therapy.

341. 341. The method of any one of claims 337-340, wherein the repeated administration of said splicing modulator, antibody-drug conjugate, or composition is concurrent with the repeated administration of said at least one additional therapy.

342. The method of any one of claims 337-340, wherein the repeated administration of said splicing modulator, antibody-drug conjugate, or composition is sequential or staggered with the repeated administration of said at least one additional therapy.

343. 343. The method of any one of claims 325-342, wherein the at least one additional therapy comprises administering a checkpoint inhibitor.

344. The method of claim 343, wherein the subject is intolerant, refractory, or non-responsive to the checkpoint inhibitor when administered alone.

345. The method of claim 343 or 344, wherein the checkpoint inhibitor targets CTLA4, PD1, PDL1, OX40, CD40, GITR, LAG3, TIM3, and / or KIR.

346. The method of any one of claims 343-345, wherein the checkpoint inhibitor targets CTLA4, OX40, CD40, and / or GITR.

347. 347. The method of any one of claims 343-346, wherein the checkpoint inhibitor comprises a cytotoxic T-lymphocyte-associated antigen 4 pathway (CTLA4) inhibitor.

348. The method of claim 347, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

349. The method of claim 348, wherein the anti-CTLA4 antibody is ipilimumab.

350. 347. The method of any one of claims 343-346, wherein the checkpoint inhibitor comprises a programmed death-1 pathway (PD1) inhibitor.

351. The method of claim 350, wherein the PD1 inhibitor is an anti-PD1 antibody.

352. The method of claim 351, wherein the anti-PD1 antibody is nivolumab.

353. The method of claim 350, wherein the PD1 inhibitor is an anti-PDL1 antibody.

354. The method of claim 353, wherein the anti-PDL1 antibody is atezolizumab.

355. The method of any one of claims 343 to 346, wherein the checkpoint inhibitors comprise a CTLA4 inhibitor and a PD1 inhibitor.

356. The method of claim 355, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

357. The method of claim 356, wherein the anti-CTLA4 antibody is ipilimumab.

358. The method of claim 355, wherein the PD1 inhibitor is an anti-PD1 antibody.

359. The method of claim 358, wherein the anti-PD1 antibody is nivolumab.

360. The method of claim 355, wherein the PD1 inhibitor is an anti-PDL1 antibody.

361. The method of claim 360, wherein the anti-PDL1 antibody is atezolizumab.

362. 343. The method of any one of claims 325-342, wherein said at least one additional therapy comprises administering a neoantigen vaccine.

363. 363. The method of claim 362, wherein said splicing modulator, antibody-drug conjugate, or composition is administered prior to administration of said neo-antigen vaccine.

364. 363. The method of claim 362, wherein said splicing modulator, antibody-drug conjugate, or composition is administered after administration of said neo-antigen vaccine.

365. 363. The method of claim 362, wherein said splicing modulator, antibody-drug conjugate, or composition is administered simultaneously with administration of said neo-antigen vaccine.

366. 366. The method of any one of claims 362-365, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is repeated at least once after an initial administration.

367. The method of claim 366, wherein the amount used for repeat administrations of the splicing modulator, antibody-drug conjugate, or composition is reduced when compared to the amount used for the initial administration.

368. 368. The method of any one of claims 362-367, wherein the neo-antigen vaccine comprises at least one neo-antigen peptide.

369. 369. The method of claim 368, wherein said at least one neo-antigenic peptide ranges from about 10 to about 50 or from about 10 to about 35 amino acids in length.

370. 370. The method of claim 368 or 369, wherein said at least one neo-antigenic peptide ranges from about 15 to about 25 amino acids in length.

371. 371. The method of any one of claims 368-370, wherein said at least one neo-antigen peptide comprises one or more neo-antigen sequences.

372. The method of claim 371, wherein the neoantigen sequence is a neoantigen sequence specific to the subject.

373. The method of claim 371, wherein the neoantigen sequence is a universal neoantigen sequence.

374. The method of any one of claims 371 to 373, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 37 to 65.

375. The method of claim 374, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

37.

376. The method of claim 374, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

39.

377. 375. The method of claim 374, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

378. 374. The method of any one of claims 371 to 373, wherein the neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 66 to 93, or an antigenic portion of any one of SEQ ID NOs: 66 to 93.

379. The method of claim 378, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO: 66, or an antigenic portion of SEQ ID NO:

66.

380. 379. The method of claim 378, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

381. 381. The method of any one of claims 374-380, wherein the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

382. 382. The method of any one of claims 374 to 381, wherein the neo-antigen sequences and / or antigenic portions are a neo-antigen vaccine personalized for the subject.

383. The method of claim 382, ​​wherein the neo-antigen sequence and / or antigenic portion is identified by sequencing at least one neo-antigen induced in the subject by administering an effective amount of the splicing modulator, antibody-drug conjugate, or composition.

384. The method of claim 382 or 383, wherein the neo-antigen sequence and / or antigenic portion has the ability to bind to at least one HLA allele expressed in the subject.

385. 382. The method of any one of claims 374 to 381, wherein said neo-antigen sequences and / or antigenic moieties are a universal neo-antigen vaccine.

386. The method of claim 385, wherein the neoantigenic sequence and / or antigenic portion is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from the neoplastic disorder.

387. The method of claim 385 or 386, wherein the neo-antigenic sequence and / or antigenic portion is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of a population of subjects suffering from the neoplastic disorder.

388. The method of any one of claims 385 to 387, wherein said at least one neo-antigen peptide comprises a neo-antigen sequence induced by contacting a neoplastic cell with an effective amount of said splicing modulator, antibody-drug conjugate, or composition.

389. 368. The method of any one of claims 362-367, wherein the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable carrier.

390. 390. The method of claim 389, wherein said at least one neo-antigenic peptide is linked to said pharmaceutically acceptable carrier.

391. The method of claim 389 or 390, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole limpet hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

392. 392. The method of any one of claims 389 to 391, wherein the neo-antigenic peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

393. 392. The method of any one of claims 389-391, wherein the neo-antigenic peptide and the pharmaceutically acceptable carrier are expressed as a fusion protein.

394. 368. The method of any one of claims 362-367, wherein the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable diluent.

395. 368. The method of any one of claims 362-367, wherein the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable adjuvant.

396. The method of claim 388, wherein the neoplastic cells are present in an in vitro cell culture.

397. The method of claim 396, wherein the neoplastic cells are obtained from the subject.

398. The method of claim 388, wherein the neoplastic cells are present in the subject.

399. 368. The method of any one of claims 362-367, wherein said neo-antigen vaccine comprises at least one neo-antigen mRNA.

400. 400. The method of claim 399, wherein said at least one neo-antigen mRNA encodes one or more neo-antigen sequences.

401. The method of claim 400, wherein the neoantigen sequence is a neoantigen sequence specific to the subject.

402. The method of claim 400, wherein the neoantigen sequence is a universal neoantigen sequence.

403. The method of any one of claims 400 to 402, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 37 to 65.

404. The method of claim 403, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

37.

405. The method of claim 403, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

39.

406. The method of claim 403, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

407. 403. The method of any one of claims 400-402, wherein the neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 66-93, or an antigenic portion of any one of SEQ ID NOs: 66-93.

408. The method of claim 407, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO: 66, or an antigenic portion of SEQ ID NO:

66.

409. 408. The method of claim 407, wherein the neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

410. 410. The method of any one of claims 403-409, wherein the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

411. 411. The method of any one of claims 403 to 410, wherein the neo-antigen sequences and / or antigenic portions are a neo-antigen vaccine personalized for the subject.

412. The method of claim 411, wherein the neo-antigen sequence and / or antigenic portion is identified by sequencing at least one neo-antigen induced in the subject by administering an effective amount of the splicing modulator, antibody-drug conjugate, or composition.

413. 413. The method of claim 411 or 412, wherein the neo-antigen sequence and / or antigenic portion has the ability to bind to at least one HLA allele expressed in the subject.

414. 411. The method of any one of claims 403 to 410, wherein said neo-antigen sequences and / or antigenic moieties are a universal neo-antigen vaccine.

415. 415. The method of claim 414, wherein the neoantigenic sequence and / or antigenic portion is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from the neoplastic disorder.

416. 416. The method of claim 414 or 415, wherein the neo-antigenic sequence and / or antigenic portion is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of a population of subjects suffering from the neoplastic disorder.

417. 417. The method of any one of claims 414-416, wherein said at least one neo-antigen mRNA encodes a neo-antigen sequence that is induced by contacting a neoplastic cell with an effective amount of said splicing modulator, antibody-drug conjugate, or composition.

418. 368. The method of any one of claims 362-367, wherein said neo-antigen vaccine comprises at least one neo-antigen mRNA and a pharmaceutically acceptable carrier.

419. 419. The method of claim 418, wherein said at least one neo-antigen mRNA is linked to said pharmaceutically acceptable carrier.

420. 420. The method of claim 418 or 419, wherein the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine.

421. 368. The method of any one of claims 362-367, wherein said neo-antigen vaccine comprises at least one neo-antigen mRNA and a pharmaceutically acceptable diluent.

422. 368. The method of any one of claims 362-367, wherein said neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable adjuvant.

423. 423. The method of any one of claims 399-422, wherein the neo-antigen mRNA is encapsulated by an encapsulating agent.

424. The method of claim 423, wherein the encapsulating agent is a liposome.

425. 424. The method of claim 423, wherein the encapsulating agent is a nanoparticle.

426. The method of claim 417, wherein the neoplastic cells are present in an in vitro cell culture.

427. The method of claim 426, wherein the neoplastic cells are obtained from the subject.

428. The method of claim 417, wherein the neoplastic cells are present in the subject.

429. 343. The method of any one of claims 325-342, wherein the at least one additional therapy comprises administering a cytokine or cytokine analog.

430. The method of claim 429, wherein the subject is intolerant, refractory, or poorly responsive to the cytokine or cytokine analog when administered alone.

431. 431. The method of claim 429 or 430, wherein the cytokine or cytokine analog comprises a T cell enhancer.

432. 432. The method of any one of claims 429-431, wherein the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, IL-15, IFNγ, and / or TNFα.

433. 343. The method of any one of claims 325-342, wherein said at least one additional therapy comprises administering modified tumor-targeting T cells.

434. The method of any one of claims 315 to 433, further comprising detecting one or more neo-antigens and / or T cell responses in the subject following administration of the splicing modulator, antibody-drug conjugate, or composition.

435. The method of claim 434, further comprising continuing administration of said splicing modulator, antibody-drug conjugate, or composition if one or more neo-antigen and / or T cell responses are detected.

436. The method of claim 434 or 435, further comprising continuing administration of the splicing modulator, antibody-drug conjugate, or composition less frequently and / or at a reduced dosage if one or more neo-antigen and / or T cell responses are detected.

437. The method of any one of claims 434 to 436, wherein detection of one or more neoantigens and / or T cell responses in the subject indicates the effectiveness of treatment with the splicing modulator, antibody-drug conjugate, or composition.

438. The method of any one of claims 315-437, wherein said subject has a non-synonymous mutation load of about 150 mutations or less.

439. The method of any one of claims 315-438, wherein said subject has a non-synonymous mutation load of about 100 mutations or less.

440. The method of any one of claims 315-439, wherein said subject has a nonsynonymous mutation load of about 50 mutations or less.

441. 441. The method of any one of claims 315 to 440, wherein the neoplastic disorder is a hematological malignancy or a solid tumor.

442. 442. The method of claim 441, wherein the hematological malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

443. 443. The method of claim 441 or 442, wherein the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma.

444. 442. The method of claim 441, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, cervical cancer, pancreatic cancer, renal cancer, colorectal cancer, and esophageal cancer.

445. 1. A method of treating a subject having or suspected of having a neoplastic disorder, comprising: (a) administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1-193, or a composition of any one of claims 194-197, wherein administration of the splicing modulator, antibody-drug conjugate, or composition induces at least one neo-antigen and / or T cell response; (b) detecting one or more neo-antigens and / or T cell responses in the subject after administration of the splicing modulator, antibody-drug conjugate, or composition; and (c) continuing the administration of the splicing modulator, antibody-drug conjugate, or composition if one or more neo-antigen and / or T cell responses are detected. A method comprising:

446. The method of claim 445, wherein detection of one or more neoantigens and / or T cell responses in the subject indicates the effectiveness of treatment with the splicing modulator, antibody-drug conjugate, or composition.

447. The method of claim 445 or 446, wherein the one or more neoantigens comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

448. The method of claim 447, wherein said one or more neoantigens comprises the amino acid sequence of SEQ ID NO:

37.

449. The method of claim 447, wherein said one or more neoantigens comprises the amino acid sequence of SEQ ID NO:

39.

450. The method of claim 447, wherein said one or more neoantigens comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

451. 197. A method of treating a subject having or suspected of having a neoplastic disorder, comprising administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1-193, or a composition of any one of claims 194-197; and at least one additional therapy.

452. 452. The method of claim 451, wherein the at least one additional therapy comprises at least 1, at least 2, at least 3, at least 4, or at least 5 additional therapies.

453. The method of claim 451 or 452, wherein administration of the splicing modulator, antibody-drug conjugate, or composition induces at least one neo-antigen and / or T cell response.

454. The method of any one of claims 451 to 453, wherein the dosage of the splicing modulator, antibody-drug conjugate or composition and / or the at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to standard dosages of the splicing modulator, antibody-drug conjugate or composition and / or the at least one additional therapy.

455. The method of any one of claims 451 to 454, wherein the splicing modulator, antibody-drug conjugate or composition and / or the at least one additional therapy is administered at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% less frequently when compared to a standard administration regimen of the splicing modulator, antibody-drug conjugate or composition and / or the at least one additional therapy.

456. The method of any one of claims 451 to 455, wherein the administration amount and / or dosage of said splicing modulator, antibody-drug conjugate or composition and / or said at least one additional therapy results in reduced systemic toxicity and / or improved tolerability.

457. The method of any one of claims 451-456, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is initiated prior to administration of the at least one additional therapy.

458. The method of any one of claims 451 to 456, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is initiated after administration of the at least one additional therapy.

459. The method of any one of claims 451-456, wherein administration of said splicing modulator, antibody-drug conjugate, or composition is commenced simultaneously with administration of said at least one additional therapy.

460. The method of any one of claims 451 to 460, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is repeated at least once after an initial administration.

461. The method of claim 460, wherein the amount used for repeat administrations of the splicing modulator, antibody-drug conjugate, or composition is reduced when compared to the amount used for the initial administration.

462. The method of claim 460 or 461, wherein the amount used for repeated administration of the splicing modulator, antibody-drug conjugate, or composition is reduced when compared to a standard dosage of the splicing modulator, antibody-drug conjugate, or composition.

463. The method of any one of claims 460 to 462, wherein the amount used for repeat administration of the splicing modulator, antibody-drug conjugate, or composition is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to a standard dosage of the splicing modulator, antibody-drug conjugate, or composition.

464. 464. The method of any one of claims 451-463, wherein the administration of the at least one additional therapy is repeated at least once after an initial administration.

465. The method of claim 464, wherein the amount used in repeat administrations of the at least one additional therapy is reduced when compared to the amount used in the initial administration.

466. 466. The method of claim 464 or 465, wherein the amount used for repeat administration of the at least one additional therapy is reduced when compared to the standard dosage of the at least one additional therapy.

467. 467. The method of any one of claims 464-466, wherein the amount used for repeat administration of the at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to a standard dosage of the at least one additional therapy.

468. The method of any one of claims 464-467, wherein the repeated administration of said splicing modulator, antibody-drug conjugate, or composition is concurrent with the repeated administration of said at least one additional therapy.

469. The method of any one of claims 464-467, wherein the repeated administration of said splicing modulator, antibody-drug conjugate, or composition is sequential or staggered with the repeated administration of said at least one additional therapy.

470. 470. The method of any one of claims 451-469, wherein said at least one additional therapy comprises administering a checkpoint inhibitor.

471. The method of claim 470, wherein the subject is intolerant, refractory, or non-responsive to the checkpoint inhibitor when administered alone.

472. The method of claim 470 or 471, wherein the checkpoint inhibitor targets CTLA4, PD1, PDL1, OX40, CD40, GITR, LAG3, TIM3, and / or KIR.

473. The method of any one of claims 470-472, wherein the checkpoint inhibitor targets CTLA4, OX40, CD40, and / or GITR.

474. 474. The method of any one of claims 470-473, wherein said checkpoint inhibitor comprises a cytotoxic T-lymphocyte-associated antigen 4 pathway (CTLA4) inhibitor.

475. The method of claim 474, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

476. The method of claim 475, wherein the anti-CTLA4 antibody is ipilimumab.

477. 474. The method of any one of claims 470-473, wherein the checkpoint inhibitor comprises a programmed death-1 pathway (PD1) inhibitor.

478. The method of claim 477, wherein the PD1 inhibitor is an anti-PD1 antibody.

479. The method of claim 478, wherein the anti-PD1 antibody is nivolumab.

480. The method of claim 477, wherein the PD1 inhibitor is an anti-PDL1 antibody.

481. The method of claim 480, wherein the anti-PDL1 antibody is atezolizumab.

482. The method of any one of claims 470 to 473, wherein the checkpoint inhibitors comprise a CTLA4 inhibitor and a PD1 inhibitor.

483. The method of claim 482, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

484. The method of claim 483, wherein the anti-CTLA4 antibody is ipilimumab.

485. The method of claim 482, wherein the PD1 inhibitor is an anti-PD1 antibody.

486. The method of claim 485, wherein the anti-PD1 antibody is nivolumab.

487. The method of claim 482, wherein the PD1 inhibitor is an anti-PDL1 antibody.

488. The method of claim 487, wherein the anti-PDL1 antibody is atezolizumab.

489. 470. The method of any one of claims 451-469, wherein said at least one additional therapy comprises administering a neoantigen vaccine.

490. 490. The method of claim 489, wherein the splicing modulator, antibody-drug conjugate, or composition is administered prior to administration of the neo-antigen vaccine.

491. 490. The method of claim 489, wherein said splicing modulator, antibody-drug conjugate, or composition is administered after administration of said neo-antigen vaccine.

492. 490. The method of claim 489, wherein said splicing modulator, antibody-drug conjugate, or composition is administered simultaneously with administration of said neoantigen vaccine.

493. 493. The method of any one of claims 489 to 492, wherein administration of the splicing modulator, antibody-drug conjugate, or composition is repeated at least once after an initial administration.

494. The method of claim 493, wherein the amount used for repeat administrations of the splicing modulator, antibody-drug conjugate, or composition is reduced when compared to the amount used for the initial administration.

495. 495. The method of any one of claims 489-494, wherein the neo-antigen vaccine comprises at least one neo-antigen peptide.

496. 496. The method of claim 495, wherein said at least one neo-antigenic peptide ranges from about 10 to about 50 or from about 10 to about 35 amino acids in length.

497. 497. The method of claim 495 or 496, wherein said at least one neo-antigenic peptide ranges from about 15 to about 25 amino acids in length.

498. 498. The method of any one of claims 495 to 497, wherein said at least one neo-antigen peptide comprises one or more neo-antigen sequences.

499. The method of claim 498, wherein the neoantigen sequence is a neoantigen sequence specific to the subject.

500. The method of claim 498, wherein the neoantigen sequence is a universal neoantigen sequence.

501. The method of any one of claims 498-500, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

502. The method of claim 501, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

37.

503. The method of claim 501, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

39.

504. 502. The method of claim 501, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

505. 501. The method of any one of claims 498-500, wherein the neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 66-93, or an antigenic portion of any one of SEQ ID NOs: 66-93.

506. The method of claim 505, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO: 66, or an antigenic portion of SEQ ID NO:

66.

507. 506. The method of claim 505, wherein the neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

508. 508. The method of any one of claims 501-507, wherein the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

509. 509. The method of any one of claims 501 to 508, wherein said neo-antigen sequences and / or antigenic portions are a neo-antigen vaccine personalized for said subject.

510. The method of claim 509, wherein said neo-antigen sequence and / or antigenic portion is identified by sequencing at least one neo-antigen induced in said subject by administering an effective amount of said splicing modulator, antibody-drug conjugate, or composition.

511. The method of claim 509 or 510, wherein the neo-antigen sequence and / or antigenic portion has the ability to bind to at least one HLA allele expressed in the subject.

512. 509. The method of any one of claims 501 to 508, wherein said neo-antigen sequences and / or antigenic moieties are a universal neo-antigen vaccine.

513. 513. The method of claim 512, wherein said neo-antigenic sequence and / or antigenic portion is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from said neoplastic disorder.

514. The method of claim 512 or 513, wherein the neo-antigenic sequence and / or antigenic portion is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of a population of subjects suffering from the neoplastic disorder.

515. 515. The method of any one of claims 512-514, wherein said at least one neo-antigen peptide comprises a neo-antigen sequence induced by contacting a neoplastic cell with an effective amount of said splicing modulator, antibody-drug conjugate, or composition.

516. 495. The method of any one of claims 489-494, wherein the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable carrier.

517. 517. The method of claim 516, wherein said at least one neo-antigenic peptide is linked to said pharmaceutically acceptable carrier.

518. The method of claim 516 or 517, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole limpet hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

519. 519. The method of any one of claims 516 to 518, wherein the neo-antigenic peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

520. 519. The method of any one of claims 516 to 518, wherein the neo-antigenic peptide and the pharmaceutically acceptable carrier are expressed as a fusion protein.

521. 495. The method of any one of claims 489-494, wherein the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable diluent.

522. 495. The method of any one of claims 489-494, wherein the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable adjuvant.

523. The method of claim 515, wherein the neoplastic cells are present in an in vitro cell culture.

524. The method of claim 523, wherein the neoplastic cells are obtained from the subject.

525. The method of claim 515, wherein the neoplastic cells are present in the subject.

526. 495. The method of any one of claims 489-494, wherein said neo-antigen vaccine comprises at least one neo-antigen mRNA.

527. 527. The method of claim 526, wherein said at least one neo-antigen mRNA encodes one or more neo-antigen sequences.

528. The method of claim 527, wherein the neoantigen sequence is a neoantigen sequence specific to the subject.

529. The method of claim 527, wherein the neoantigen sequence is a universal neoantigen sequence.

530. 530. The method of any one of claims 527 to 529, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 37 to 65.

531. The method of claim 530, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

37.

532. The method of claim 530, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

39.

533. 531. The method of claim 530, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

534. 530. The method of any one of claims 527-529, wherein the neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 66-93, or an antigenic portion of any one of SEQ ID NOs: 66-93.

535. 535. The method of claim 534, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO: 66, or an antigenic portion of SEQ ID NO:

66.

536. 535. The method of claim 534, wherein the neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

537. 537. The method of any one of claims 530-536, wherein the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

538. 538. The method of any one of claims 530 to 537, wherein the neo-antigen sequences and / or antigenic portions are a neo-antigen vaccine personalized for the subject.

539. The method of claim 538, wherein the neo-antigen sequence and / or antigenic portion is identified by sequencing at least one neo-antigen induced in the subject by administering an effective amount of the splicing modulator, antibody-drug conjugate, or composition.

540. The method of claim 538 or 539, wherein the neo-antigen sequence and / or antigenic portion has the ability to bind to at least one HLA allele expressed in the subject.

541. 538. The method of any one of claims 530 to 537, wherein the neo-antigen sequences and / or antigenic moieties are a universal neo-antigen vaccine.

542. 542. The method of claim 541, wherein the neoantigenic sequence and / or antigenic portion is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from the neoplastic disorder.

543. The method of claim 541 or 542, wherein the neo-antigenic sequence and / or antigenic portion is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of a population of subjects suffering from the neoplastic disorder.

544. 544. The method of any one of claims 541-543, wherein said at least one neo-antigen mRNA encodes a neo-antigen sequence that is induced by contacting a neoplastic cell with an effective amount of said splicing modulator, antibody-drug conjugate, or composition.

545. 495. The method of any one of claims 489-494, wherein said neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable carrier.

546. The method of claim 545, wherein said at least one neo-antigen mRNA is linked to said pharmaceutically acceptable carrier.

547. The method of claim 545 or 546, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole limpet hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

548. 495. The method of any one of claims 489-494, wherein said neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable diluent.

549. 495. The method of any one of claims 489-494, wherein said neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable adjuvant.

550. 550. The method of any one of claims 526 to 549, wherein the neo-antigen mRNA is encapsulated by an encapsulating agent.

551. The method of claim 550, wherein the encapsulating agent is a liposome.

552. The method of claim 550, wherein the encapsulating agent is a nanoparticle.

553. The method of claim 544, wherein the neoplastic cells are present in an in vitro cell culture.

554. The method of claim 553, wherein the neoplastic cells are obtained from the subject.

555. The method of claim 544, wherein the neoplastic cells are present in the subject.

556. 470. The method of any one of claims 451-469, wherein the at least one additional therapy comprises administering a cytokine or cytokine analog.

557. The method of claim 556, wherein the subject is intolerant, refractory, or poorly responsive to the cytokine or cytokine analog when administered alone.

558. The method of claim 556 or 557, wherein the cytokine or cytokine analog comprises a T cell enhancer.

559. 559. The method of any one of claims 556 to 558, wherein the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, IL-15, IFNγ, and / or TNFα.

560. 470. The method of any one of claims 451-469, wherein said at least one additional therapy comprises administering modified tumor-targeting T cells.

561. 561. The method of any one of claims 451-560, wherein said subject has a non-synonymous mutation load of about 150 mutations or less.

562. 562. The method of any one of claims 451-561, wherein said subject has a nonsynonymous mutation load of about 100 mutations or less.

563. 563. The method of any one of claims 451-562, wherein said subject has a nonsynonymous mutation load of about 50 mutations or less.

564. 564. The method of any one of claims 451-563, wherein the neoplastic disorder is a hematological malignancy or a solid tumor.

565. 565. The method of claim 564, wherein the hematological malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

566. The method of claim 564 or 565, wherein the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma.

567. 565. The method of claim 564, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, cervical cancer, pancreatic cancer, renal cancer, colorectal cancer, and esophageal cancer.

568. 1. A method for identifying at least one neoantigen, comprising: (a) contacting neoplastic cells with an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1-193, or a composition of any one of claims 194-197; (b) detecting at least one alternatively spliced ​​mRNA transcript after contacting the neoplastic cell with the splicing modulator, antibody-drug conjugate, or composition; (c) predicting the translation of said at least one alternatively spliced ​​mRNA transcript into at least one peptide; and (d) comparing the at least one peptide to a reference proteome, wherein if the at least one peptide does not match any peptide in the reference proteome, at least one neoantigen is identified. A method comprising:

569. 569. The method of claim 568, wherein detecting at least one alternatively spliced ​​mRNA transcript comprises RNAseq.

570. 569. The method of claim 568 or 569, wherein predicting the translation of the at least one alternatively spliced ​​mRNA transcript comprises quantifying the change in percent spliced-in (dPSI) value for the at least one transcript.

571. 571. The method of any one of claims 568-570, wherein predicting translation of said at least one alternatively spliced ​​mRNA transcript comprises RiboSeq and / or ribosome profiling.

572. 572. The method of any one of claims 568-571, further comprising evaluating said at least one peptide for predicted major histocompatibility complex (MHC) binding.

573. 573. The method of claim 572, wherein the predicted MHC binding is determined by measuring the uncorrected affinity predicted binding strength of said at least one peptide.

574. The method of claim 573, wherein an uncorrected affinity predicted binding strength of about 500 nM or greater indicates MHC binding.

575. 573. The method of claim 572, wherein the predicted MHC binding is determined by identifying a distribution of predicted binding strengths for a set of random peptides; and comparing the predicted binding strength of said at least one peptide to said distribution.

576. 576. The method of claim 575, wherein predicted binding strengths in the top 2.0% of the distribution indicate weak MHC binding.

577. 576. The method of claim 575, wherein predicted binding strengths in the top 0.5% of the distribution indicate strong MHC binding.

578. The method of any one of claims 568 to 577, wherein the neoplastic cells are in an in vitro cell culture.

579. The method of claim 578, wherein the neoplastic cells are obtained from the subject.

580. The method of any one of claims 568 to 577, wherein the neoplastic cells are present in the subject.

581. 581. The method of any one of claims 568-580, further comprising contacting one or more additional neoplastic cells to identify at least one universal neoantigen.

582. 1. A method for identifying at least one neoantigen, comprising: (a) contacting neoplastic cells with an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1-193, or a composition of any one of claims 194-197; (b) detecting at least one peptide comprising a potential neoantigenic sequence after contacting the neoplastic cells with the splicing modulator, antibody-drug conjugate, or composition; and (c) comparing the at least one peptide to a reference proteome, wherein if the at least one peptide does not match any peptide in the reference proteome, at least one neoantigen is identified. A method comprising:

583. 583. The method of claim 582, further comprising evaluating the at least one peptide for predicted major histocompatibility complex (MHC) binding.

584. 584. The method of claim 583, wherein the predicted MHC binding is determined by measuring the uncorrected affinity predicted binding strength of said at least one peptide.

585. The method of claim 584, wherein an uncorrected affinity predicted binding strength of about 500 nM or greater indicates MHC binding.

586. 584. The method of claim 583, wherein the predicted MHC binding is determined by identifying a distribution of predicted binding strengths for a set of random peptides; and comparing the predicted binding strength of said at least one peptide to said distribution.

587. The method of claim 586, wherein predicted binding strengths in the top 2.0% of the distribution indicate weak MHC binding.

588. The method of claim 586, wherein predicted binding strengths in the top 0.5% of the distribution indicate strong MHC binding.

589. The method of any one of claims 582 to 588, wherein the neoplastic cells are in in vitro cell culture.

590. The method of claim 589, wherein the neoplastic cells are obtained from the subject.

591. The method of any one of claims 582 to 588, wherein the neoplastic cells are present in the subject.

592. 592. The method of any one of claims 582-591, further comprising contacting one or more additional neoplastic cells to identify at least one universal neoantigen.

593. 1. A method for making a neo-antigen vaccine, comprising: (a) identifying at least one neoantigen using the method of any one of claims 568 to 592; and (b) formulating said at least one neoantigen with a pharmaceutically acceptable carrier, diluent, or adjuvant; A method comprising:

594. 594. The method of claim 593, wherein said at least one neoantigen comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

595. 595. The method of claim 594, wherein said at least one neoantigen comprises the amino acid sequence of SEQ ID NO:

37.

596. 595. The method of claim 594, wherein said at least one neoantigen comprises the amino acid sequence of SEQ ID NO:

39.

597. 595. The method of claim 594, wherein said at least one neo-antigen comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

598. 594. The method of claim 593, wherein said at least one neo-antigen comprises the amino acid sequence of any one of SEQ ID NOs: 66-93, or an antigenic portion of any one of SEQ ID NOs: 66-93.

599. 599. The method of claim 598, wherein said at least one neo-antigen comprises the amino acid sequence of SEQ ID NO: 66, or an antigenic portion of SEQ ID NO:

66.

600. 600. The method of claim 598, wherein said at least one neo-antigen comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

601. 601. The method of any one of claims 593-600, wherein said at least one neo-antigen and / or antigenic portion ranges from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

602. 602. The method of any one of claims 593 to 601, wherein said at least one neo-antigen and / or antigenic moiety is linked to said pharmaceutically acceptable carrier.

603. The method of any one of claims 593 to 602, wherein the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine.

604. 1. A method of treating a subject having or suspected of having a neoplastic disorder, comprising: (a) administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1-193, or a composition of any one of claims 194-197; (b) detecting one or more neoantigens in the subject after administration of the splicing modulator, antibody-drug conjugate, or composition; (c) comparing said one or more neoantigens to a panel of universal neoantigens; and (d) administering to the subject a universal neo-antigen vaccine comprising at least one universal neo-antigen present in the subject. A method comprising:

605. 605. The method of claim 604, wherein the universal neo-antigen vaccine is administered alone or in combination with at least one additional therapy.

606. The method of claim 605, wherein the at least one additional therapy comprises at least 1, at least 2, at least 3, at least 4, or at least 5 additional therapies.

607. The method of claim 605 or 606, wherein said at least one additional therapy comprises repeated administration of said splicing modulator, antibody-drug conjugate, or composition.

608. The method of claim 607, wherein the repeated administration of said splicing modulator, antibody-drug conjugate, or composition is initiated prior to administration of said universal neo-antigen vaccine.

609. The method of claim 607, wherein repetition of the splicing modulator, antibody-drug conjugate, or composition is initiated after administration of the universal neo-antigen vaccine.

610. The method of claim 607, wherein the repeated administration of said splicing modulator, antibody-drug conjugate, or composition is initiated simultaneously with administration of said universal neo-antigen vaccine.

611. The method of any one of claims 607 to 610, wherein the amount used in repeat administrations of the splicing modulator, antibody-drug conjugate, or composition is reduced when compared to the amount used in the initial administration.

612. The method of any one of claims 607 to 611, wherein the amount used for repeated administration of the splicing modulator, antibody-drug conjugate, or composition is reduced when compared to a standard dosage of the splicing modulator, antibody-drug conjugate, or composition.

613. The method of any one of claims 607 to 612, wherein the amount used for repeat administration of the splicing modulator, antibody-drug conjugate, or composition is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to a standard dosage of the splicing modulator, antibody-drug conjugate, or composition.

614. 614. The method of any one of claims 605-613, wherein the at least one additional therapy comprises administering a checkpoint inhibitor.

615. 615. The method of claim 614, wherein administration of said checkpoint inhibitor is initiated prior to administration of said universal neo-antigen vaccine and / or repeated administration of said splicing modulator, antibody-drug conjugate or composition.

616. 615. The method of claim 614, wherein administration of said checkpoint inhibitor is initiated after repeat administration of said universal neo-antigen vaccine and / or said splicing modulator, antibody-drug conjugate or composition.

617. 615. The method of claim 614, wherein administration of said checkpoint inhibitor is initiated simultaneously with administration of said universal neo-antigen vaccine and / or repeated administration of said splicing modulator, antibody-drug conjugate or composition.

618. 618. The method of any one of claims 614-617, wherein administration of the checkpoint inhibitor is repeated at least one time after an initial administration.

619. The method of claim 618, wherein the amount used for repeat administration of the checkpoint inhibitor is reduced as compared to the amount used for the initial administration.

620. 620. The method of claim 618 or 619, wherein the amount used for repeat administration of the checkpoint inhibitor is reduced when compared to a standard dosage of the checkpoint inhibitor.

621. 621. The method of any one of claims 618-620, wherein the amount used for repeat administration of the checkpoint inhibitor is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to a standard dosage of the checkpoint inhibitor.

622. The method of any one of claims 614 to 621, wherein the subject is intolerant, refractory, or non-responsive to the checkpoint inhibitor when administered alone.

623. 623. The method of any one of claims 614-622, wherein the checkpoint inhibitor targets CTLA4, PD1, PDL1, OX40, CD40, GITR, LAG3, TIM3, and / or KIR.

624. The method of any one of claims 614-623, wherein the checkpoint inhibitor targets CTLA4, OX40, CD40, and / or GITR.

625. 625. The method of any one of claims 614-624, wherein said checkpoint inhibitor comprises a cytotoxic T-lymphocyte-associated antigen 4 pathway (CTLA4) inhibitor.

626. The method of claim 625, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

627. The method of claim 626, wherein the anti-CTLA4 antibody is ipilimumab.

628. 625. The method of any one of claims 614-624, wherein the checkpoint inhibitor comprises a programmed death-1 pathway (PD1) inhibitor.

629. The method of claim 628, wherein the PD1 inhibitor is an anti-PD1 antibody.

630. The method of claim 629, wherein the anti-PD1 antibody is nivolumab.

631. The method of claim 628, wherein the PD1 inhibitor is an anti-PDL1 antibody.

632. The method of claim 631, wherein the anti-PDL1 antibody is atezolizumab.

633. The method of any one of claims 614 to 624, wherein the checkpoint inhibitors comprise a CTLA4 inhibitor and a PD1 inhibitor.

634. The method of claim 633, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

635. The method of claim 634, wherein the anti-CTLA4 antibody is ipilimumab.

636. The method of claim 633, wherein the PD1 inhibitor is an anti-PD1 antibody.

637. The method of claim 636, wherein the anti-PD1 antibody is nivolumab.

638. The method of claim 633, wherein the PD1 inhibitor is an anti-PDL1 antibody.

639. The method of claim 638, wherein the anti-PDL1 antibody is atezolizumab.

640. 640. The method of any one of claims 604-639, wherein said universal neo-antigen vaccine comprises at least one neo-antigen peptide.

641. 641. The method of claim 640, wherein said at least one neo-antigenic peptide ranges from about 10 to about 50 or from about 10 to about 35 amino acids in length.

642. 642. The method of claim 640 or 641, wherein said at least one neo-antigenic peptide ranges from about 15 to about 25 amino acids in length.

643. 643. The method of any one of claims 640-642, wherein said at least one neo-antigen peptide comprises one or more universal neo-antigen sequences.

644. The method of claim 643, wherein the universal neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

645. The method of claim 644, wherein the universal neo-antigen sequence comprises the amino acid sequence of SEQ ID NO:

37.

646. The method of claim 644, wherein the universal neo-antigen sequence comprises the amino acid sequence of SEQ ID NO:

39.

647. The method of claim 644, wherein the universal neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

648. 644. The method of claim 643, wherein the universal neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 66-93, or an antigenic portion of any one of SEQ ID NOs: 66-93.

649. The method of claim 648, wherein the universal neo-antigen sequence comprises the amino acid sequence of SEQ ID NO: 66, or an antigenic portion of SEQ ID NO:

66.

650. 649. The method of claim 648, wherein the universal neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

651. 651. The method of any one of claims 643-650, wherein the universal neo-antigen sequences and / or antigenic portions range from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

652. 652. The method of any one of claims 643 to 651, wherein said universal neo-antigenic sequence and / or antigenic portion has the ability to bind to at least one HLA allele that is expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from said neoplastic disorder.

653. 653. The method of any one of claims 643-652, wherein said universal neo-antigenic sequence and / or antigenic portion is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of a population of subjects suffering from said neoplastic disorder.

654. 640. The method of any one of claims 604-639, wherein said universal neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable carrier.

655. 655. The method of claim 654, wherein said at least one neo-antigenic peptide is linked to said pharmaceutically acceptable carrier.

656. The method of claim 654 or 655, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole limpet hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

657. 657. The method of any one of claims 654 to 656, wherein the neo-antigenic peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

658. 657. The method of any one of claims 654 to 656, wherein the neo-antigenic peptide and the pharmaceutically acceptable carrier are expressed as a fusion protein.

659. 640. The method of any one of claims 604-639, wherein said universal neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable diluent.

660. 640. The method of any one of claims 604-639, wherein said universal neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable adjuvant.

661. 640. The method of any one of claims 604-639, wherein said universal neo-antigen vaccine comprises at least one neo-antigen mRNA.

662. The method of claim 661, wherein said at least one neo-antigen mRNA encodes one or more universal neo-antigen sequences.

663. The method of claim 662, wherein the universal neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

664. The method of claim 663, wherein the universal neo-antigen sequence comprises the amino acid sequence of SEQ ID NO:

37.

665. The method of claim 663, wherein the universal neo-antigen sequence comprises the amino acid sequence of SEQ ID NO:

39.

666. The method of claim 663, wherein the universal neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

667. 663. The method of claim 662, wherein the universal neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 66-93, or an antigenic portion of any one of SEQ ID NOs: 66-93.

668. The method of claim 667, wherein the universal neo-antigen sequence comprises the amino acid sequence of SEQ ID NO: 66, or an antigenic portion of SEQ ID NO:

66.

669. 668. The method of claim 667, wherein the universal neo-antigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

670. 669. The method of any one of claims 662-669, wherein the universal neo-antigen sequences and / or antigenic portions range from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

671. 671. The method of any one of claims 662 to 670, wherein said universal neo-antigenic sequence and / or antigenic portion has the ability to bind to at least one HLA allele that is expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from said neoplastic disorder.

672. 672. The method of any one of claims 662-671, wherein said universal neo-antigenic sequence and / or antigenic portion is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of a population of subjects suffering from said neoplastic disorder.

673. 640. The method of any one of claims 604-639, wherein said universal neo-antigen vaccine comprises at least one neo-antigen mRNA and a pharmaceutically acceptable carrier.

674. The method of claim 673, wherein said at least one neoantigen mRNA is linked to said pharmaceutically acceptable carrier.

675. The method of claim 673 or 674, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole limpet hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

676. 640. The method of any one of claims 604-639, wherein said universal neo-antigen vaccine comprises at least one neo-antigen mRNA and a pharmaceutically acceptable diluent.

677. 640. The method of any one of claims 604-639, wherein said universal neo-antigen vaccine comprises at least one neo-antigen mRNA and a pharmaceutically acceptable adjuvant.

678. The method of any one of claims 661 to 677, wherein the neoantigen mRNA is encapsulated in an encapsulating agent.

679. The method of claim 678, wherein the encapsulating agent is a liposome.

680. The method of claim 678, wherein the encapsulating agent is a nanoparticle.

681. 681. The method of any one of claims 604-680, wherein said subject has a nonsynonymous mutation load of about 150 mutations or less.

682. 682. The method of any one of claims 604-681, wherein said subject has a nonsynonymous mutation load of about 100 mutations or less.

683. 683. The method of any one of claims 604-682, wherein said subject has a nonsynonymous mutation load of about 50 mutations or less.

684. 684. The method of any one of claims 604-683, wherein the neoplastic disorder is a hematological malignancy or a solid tumor.

685. The method of claim 684, wherein the hematological malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

686. The method of claim 684 or 685, wherein the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma.

687. The method of claim 684, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, cervical cancer, pancreatic cancer, renal cancer, colorectal cancer, and esophageal cancer.

688. The antibody-drug conjugate has the formula (I): Ab-(L-D) ｐ (I) (In the formula, Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is D2, D1, D4, D12, or D15; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. The method of any one of claims 303 to 687, comprising an antibody-drug conjugate of the formula:

689. The method of claim 688, wherein the antibody or antigen-binding fragment targets HER2-expressing cells.

690. The method of claim 689, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

691. The method of claim 689 or 690, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3).

692. 692. The method of any one of claims 689 to 691, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

693. The method of claim 688, wherein the antibody or antigen-binding fragment targets CD138-expressing cells.

694. The method of claim 693, wherein the antibody or antigen-binding fragment is an anti-CD138 antibody or antigen-binding fragment.

695. The method of claim 693 or 694, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) 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 (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 10 (LCDR1), SEQ ID NO: 11 (LCDR2), and SEQ ID NO: 12 (LCDR3).

696. 696. The method of any one of claims 693 to 695, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

22.

697. The method of claim 688, wherein the antibody or antigen-binding fragment targets an EPHA2-expressing cell.

698. The method of claim 697, wherein the antibody or antigen-binding fragment is an anti-EPHA2 antibody or antigen-binding fragment.

699. The method of claim 697 or 698, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 13 (HCDR1), SEQ ID NO: 14 (HCDR2), and SEQ ID NO: 15 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 16 (LCDR1), SEQ ID NO: 17 (LCDR2), and SEQ ID NO: 18 (LCDR3).

700. 699. The method of any one of claims 697 to 699, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

24.

701. The method of any one of claims 688 to 700, wherein the linker is a cleavable linker.

702. 702. The method of claim 701, wherein the cleavable linker comprises MC-Val-Cit-pABC.

703. 702. The method of claim 701, wherein said cleavable linker comprises MC-Val-Ala-pABC.

704. 702. The method of claim 701, wherein the cleavable linker comprises MC-Val-Ala-pAB.

705. The method of any one of claims 688 to 700, wherein the linker is a non-cleavable linker.

706. 706. The method of claim 705, wherein the non-cleavable linker comprises Mal-Hex.

707. The method of claim 705, wherein the non-cleavable linker comprises Mal-Et.

708. 706. The method of claim 705, wherein the non-cleavable linker comprises Mal-Et-O-Et.

709. The splicing regulator is D2: 【Transformation 30】 The method of any one of claims 688 to 708, comprising:

710. The method of claim 709, wherein the splicing modulator comprises D2 and the linker comprises MC-Val-Ala-pAB.

711. The splicing regulator is D1: 【Chemistry 31】 The method of any one of claims 688 to 708, comprising:

712. The method of claim 711, wherein the splicing modulator comprises D1 and the linker comprises MC-Val-Cit-pABC.

713. The splicing regulator has the formula (II): 【Chemistry 32】 (In the formula, R １ Is absent, hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; and R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ Alkyl groups, and -NR １５ R １６ Selected from: R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ Selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; and Z is 【Transformation 33】 Selected from: R １ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen) or a pharmaceutically acceptable salt thereof.

714. The splicing regulator has the formula (IV): 【Transformation 34】 (In the formula, R １ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; and R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ Alkyl groups, and -NR １５ R １６ Selected from: R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ is selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; R １ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen) or a pharmaceutically acceptable salt thereof.

715. The splicing regulator has the formula (VI): 【Chemistry 35】 (In the formula, R １ and R ９ are each independently hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; R ４ , R ５ , and R ８ are each independently hydrogen, a hydroxyl group, or —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; R ６ and R ７ are each independently hydrogen, -O-R １７ , -OC(=O)-R １７ , -OC(=O)-NR １５ R １６ , C １ ~C ６ alkyl group, -NR １５ R １６ and a linker; R １０ is hydrogen, C １ ~C ６ Alkyl group, —C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ Selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; and a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; R １ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , R ９ , R １０ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; R ６ and R ７ at least one of is hydrogen; R １ and R ９ cannot both exist) or a pharmaceutically acceptable salt thereof.

716. The splicing regulator has the formula (VIII): 【Transformation 36】 (In the formula, R １ Is absent, hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl group, —O—C(═O)—(C １ ~C ６ alkyl) group, and -CD ３ Selected from: R ３ is hydrogen, C １ ~C ６ Alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylamino group, C １ ~C ６ Alkylcarboxylic acid group, C １ ~C ６ Alkylhydroxy group, C ３ ~C ８ Cycloalkyl group, benzyl group, C ３ ~C ８ Heterocyclyl groups, and —O—C(═O)—(C １ ~C ６ alkyl) groups; R ４ is hydrogen, a hydroxyl group, —O—(C １ ~C ６ alkyl) group, —O—C(═O)—(C １ ~C ６ alkyl) groups, and C １ ~C ６ alkyl groups; and R １０ is selected from a 3- to 10-membered carbocycle and a 3- to 10-membered heterocycle, each of which is selected from 0 to 3 R ａ and each R ａ are independently halogen, C １ ~C ６ alkyl group, —O—(C １ ~C ６ ) alkyl group, C １ ~C ６ Alkylalkoxy group, C １ ~C ６ Alkylhydroxy group, —S(═O) ｗ -(4- to 7-membered heterocycle), 4- to 7-membered carbocycle, and 4- to 7-membered heterocycle; R １５ and R １６ are each independently hydrogen, R １７ , —C(═O)—R １７ and —C(═O)—O—R １７ is selected from: R １７ is hydrogen, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, benzyl groups, and C ３ ~C ８ heterocyclyl groups; R １ , R ３ , R ４ , R １０ , R １５ , R １６ , and R １７ are each independently a halogen, a hydroxyl group, C １ ~C ６ alkyl group, —O—(C １ ~C ６ alkyl group, —NR １５ R １６ , C ３ ~C ８ Cycloalkyl group, C １ ~C ６ Alkylhydroxy group, C １ ~C ６ Alkylalkoxy group, benzyl group, and C ３ ~C ８ substituted with 0 to 3 groups independently selected from heterocyclyl groups; Each R ａ are independently a halogen, a hydroxyl group, or —NR １５ R １６ , C １ ~C ６ Alkyl group, -(C=O)-(C １ ~C ６ alkyl) group, -(C=O)-(C １ ~C ６ alkyl)-(C ３ ~C １０ heterocyclyl group, —S(═O) ｗ -(C ３ ~C ８ heterocyclyl groups), and C １ ~C ６ alkylcarboxylic acid groups (wherein each of these is selected from the group consisting of halogen, hydroxyl, -NR １５ R １６ , and C １ ~C ３ substituted with 0 to 3 groups independently selected from alkyl; w is 0, 1, or 2. or a pharmaceutically acceptable salt thereof.

717. The method of any one of claims 688 to 708, wherein the splicing modulator comprises a modulator of the SF3b complex.

718. The method of claim 717, wherein the splicing regulator comprises pladienolide or a pladienolide derivative.

719. The method of claim 717 or 718, wherein the splicing modulator comprises pladienolide D or a pladienolide D derivative.

720. 720. The method of claim 719, wherein the pladienolide D or derivative comprises D2, D1, D4, D8, D10, D11 (E7107), D20, D21, D22, D12, or D25.

721. The method of claim 719 or 720, wherein the pladienolide D or derivative comprises D2.

722. The method of claim 719 or 720, wherein the pladienolide D or derivative comprises D1.

723. The method of claim 719 or 720, wherein the pladienolide D or derivative is a zwitterionic pladienolide D or derivative.

724. The method of claim 723, wherein the zwitterionic pladienolide D or derivative comprises D22 or D25.

725. The method of claim 717 or 718, wherein the splicing modulator comprises pladienolide B or a pladienolide B derivative.

726. The method of claim 725, wherein the pladienolide B or derivative comprises D9, D18, D19, or D13.

727. The method of claim 717 or 718, wherein the splicing modulator comprises an arylpladienolide.

728. 728. The method of claim 727, wherein the arylpladienolide comprises D15, D14, D16, D17, D26, or D33.

729. The method of claim 727 or 728, wherein the arylpladienolide is a zwitterionic arylpladienolide.

730. 730. The method of claim 729, wherein the zwitterionic arylpladienolide comprises D33.

731. 709. The method of any one of claims 688-708, wherein the splicing modulator comprises D4, D12, D15, D8, D9, D10, D13, D18, D19, D20, D21, D22, D25, or D33.

732. The method of any one of claims 688 to 731, wherein p is 2 to 8.

733. The method of any one of claims 688 to 732, wherein p is 4 to 8.

734. The method of any one of claims 688 to 733, wherein p is 4 or 8.

735. 202. A neo-antigen vaccine comprising at least one neo-antigen peptide, wherein said at least one neo-antigen peptide comprises a modified or novel neo-antigen sequence induced by contacting a neoplastic cell with an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1 to 193, or a composition of any one of claims 194 to 197.

736. 736. The neo-antigen vaccine of claim 735, wherein said at least one neo-antigen peptide ranges from about 10 to about 50 or from about 10 to about 35 amino acids in length.

737. 737. The neo-antigen vaccine of claim 735 or 736, wherein said at least one neo-antigen peptide ranges from about 15 to about 25 amino acids in length.

738. The neoantigen vaccine of any one of claims 735-737, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

739. The neoantigen vaccine of claim 738, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

37.

740. The neoantigen vaccine of claim 738, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

39.

741. The neoantigen vaccine of claim 738, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

742. 738. The neoantigen vaccine of any one of claims 735-737, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 66-93, or an antigenic portion of any one of SEQ ID NOs: 66-93.

743. The neoantigen vaccine of claim 742, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:66, or an antigenic portion of SEQ ID NO:

66.

744. 743. The neoantigen vaccine of claim 742, wherein said modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

745. 745. The neo-antigen vaccine of any one of claims 735-744, wherein the modified or novel neo-antigen sequence and / or antigenic portion ranges from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

746. 746. The neo-antigen vaccine of any one of claims 735-745, further comprising a pharmaceutically acceptable carrier.

747. The neoantigen vaccine of claim 746, wherein said at least one neoantigen peptide is linked to said pharmaceutically acceptable carrier.

748. 748. The neo-antigen vaccine of claim 746 or 747, wherein the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine.

749. The neoantigen vaccine of any one of claims 746 to 748, wherein the neoantigen peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

750. The neoantigen vaccine of any one of claims 746-748, wherein the neoantigen peptide and the pharmaceutically acceptable carrier are expressed as a fusion protein.

751. 746. The neo-antigen vaccine of any one of claims 735-745, further comprising a pharmaceutically acceptable diluent.

752. 746. The neo-antigen vaccine of any one of claims 735 to 745, further comprising a pharmaceutically acceptable adjuvant.

753. The neoantigen vaccine of any one of claims 735-752, wherein said neoplastic cells are in in vitro cell culture.

754. The neoantigen vaccine of claim 753, wherein the neoplastic cells are obtained from a subject.

755. The neoantigen vaccine of any one of claims 735-752, wherein the neoplastic cells are present in a subject.

756. 202. A neo-antigen vaccine comprising at least one neo-antigen mRNA, wherein said at least one neo-antigen mRNA encodes a modified or novel neo-antigen sequence induced by contacting a neoplastic cell with an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 1-193, or a composition of any one of claims 194-197.

757. The neoantigen vaccine of claim 756, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 37-65.

758. The neoantigen vaccine of claim 757, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

37.

759. The neoantigen vaccine of claim 757, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:

39.

760. The neoantigen vaccine of claim 757, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 46-49.

761. 757. The neoantigen vaccine of claim 756, wherein said modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 66-93, or an antigenic portion of any one of SEQ ID NOs: 66-93.

762. The neoantigen vaccine of claim 761, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:66, or an antigenic portion of SEQ ID NO:

66.

763. 762. The neoantigen vaccine of claim 761, wherein said modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NOs: 74-77, or an antigenic portion of any one of SEQ ID NOs: 74-77.

764. 764. The neo-antigen vaccine of any one of claims 756-763, wherein the modified or novel neo-antigen sequence and / or antigenic portion ranges from about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids in length.

765. 765. The neo-antigen vaccine of any one of claims 756-764, further comprising a pharmaceutically acceptable carrier.

766. The neoantigen vaccine of claim 765, wherein said at least one neoantigen mRNA is linked to said pharmaceutically acceptable carrier.

767. 767. The neo-antigen vaccine of claim 765 or 766, wherein the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine.

768. 765. The neo-antigen vaccine of any one of claims 756-764, further comprising a pharmaceutically acceptable diluent.

769. 765. The neo-antigen vaccine of any one of claims 756-764, further comprising a pharmaceutically acceptable adjuvant.

770. The neo-antigen vaccine of any one of claims 756-769, wherein the neo-antigen mRNA is encapsulated by an encapsulating agent.

771. The neo-antigen vaccine of claim 770, wherein the encapsulating agent is a liposome.

772. The neo-antigen vaccine of claim 770, wherein the encapsulating agent is a nanoparticle.

773. The neoantigen vaccine of any one of claims 756-772, wherein said neoplastic cells are present in an in vitro cell culture.

774. The neoantigen vaccine of claim 773, wherein the neoplastic cells are obtained from a subject.

775. The neoantigen vaccine of any one of claims 756-772, wherein the neoplastic cells are present in a subject.

776. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets MSLN-expressing cells.

777. The antibody-drug conjugate of claim 776, wherein the antibody or antigen-binding fragment is an anti-MSLN antibody or antigen-binding fragment.

778. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets FOLH1-expressing cells.

779. The antibody-drug conjugate of claim 778, wherein the antibody or antigen-binding fragment is an anti-FOLH1 antibody or antigen-binding fragment.

780. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets CDH6-expressing cells.

781. The antibody-drug conjugate of claim 780, wherein the antibody or antigen-binding fragment is an anti-CDH6 antibody or antigen-binding fragment.

782. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets CEACAM5-expressing cells.

783. The antibody-drug conjugate of claim 782, wherein the antibody or antigen-binding fragment is an anti-CEACAM5 antibody or antigen-binding fragment.

784. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets CFC1B-expressing cells.

785. The antibody-drug conjugate of claim 784, wherein the antibody or antigen-binding fragment is an anti-CFC1B antibody or antigen-binding fragment.

786. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets ENPP3-expressing cells.

787. The antibody-drug conjugate of claim 786, wherein the antibody or antigen-binding fragment is an anti-ENPP3 antibody or antigen-binding fragment.

788. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets FOLR1-expressing cells.

789. The antibody-drug conjugate of claim 788, wherein the antibody or antigen-binding fragment is an anti-FOLR1 antibody or antigen-binding fragment.

790. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets HAVCR1-expressing cells.

791. The antibody-drug conjugate of claim 790, wherein the antibody or antigen-binding fragment is an anti-HAVCR1 antibody or antigen-binding fragment.

792. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets KIT-expressing cells.

793. The antibody-drug conjugate of claim 792, wherein the antibody or antigen-binding fragment is an anti-KIT antibody or antigen-binding fragment.

794. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets MET-expressing cells.

795. The antibody-drug conjugate of claim 794, wherein the antibody or antigen-binding fragment is an anti-MET antibody or antigen-binding fragment.

796. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets MUC16-expressing cells.

797. The antibody-drug conjugate of claim 796, wherein the antibody or antigen-binding fragment is an anti-MUC16 antibody or antigen-binding fragment.

798. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets SLC39A6-expressing cells.

799. The antibody-drug conjugate of claim 798, wherein the antibody or antigen-binding fragment is an anti-SLC39A6 antibody or antigen-binding fragment.

800. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets SLC44A4-expressing cells.

801. The antibody-drug conjugate of claim 800, wherein the antibody or antigen-binding fragment is an anti-SLC44A4 antibody or antigen-binding fragment.

802. The antibody-drug conjugate of any one of claims 1 to 7, wherein the antibody or antigen-binding fragment targets STEAP1-expressing cells.

803. The antibody-drug conjugate of claim 802, wherein the antibody or antigen-binding fragment is an anti-STEAP1 antibody or antigen-binding fragment.

804. 30. The antibody-drug conjugate of claim 29, wherein the amino acid unit comprises glutamine-valine-citrulline (Glu-Val-Cit).

805. 30. The antibody-drug conjugate of claim 29, wherein the amino acid unit comprises alanine-alanine-asparagine (Ala-Ala-Asn).

806. 50. The antibody-drug conjugate of claim 48 or 49, wherein the cleavable peptide moiety or amino acid unit comprises Glu-Val-Cit.

807. 50. The antibody-drug conjugate of claim 48 or 49, wherein the cleavable peptide moiety or amino acid unit comprises Ala-Ala-Asn.

808. The antibody-drug conjugate of any one of claims 55 to 58, wherein the linker comprises MC-Glu-Val-Cit.

809. The antibody-drug conjugate of any one of claims 55 to 58, wherein the linker comprises MC-Ala-Ala-Asn.

810. The antibody-drug conjugate of any one of claims 67 to 71, wherein the linker comprises Glu-Val-Cit-pABC.

811. The antibody-drug conjugate of any one of claims 67 to 71, wherein the linker comprises Ala-Ala-Asn-pABC.

812. The antibody-drug conjugate of any one of claims 74 to 78, wherein the linker comprises Glu-Val-Cit-pAB.

813. The antibody-drug conjugate of any one of claims 74 to 78, wherein the linker comprises Ala-Ala-Asn-pAB.

814. The antibody-drug conjugate of claim 117 or 118, wherein the pladienolide D or derivative comprises D4.

815. The antibody-drug conjugate of claim 117 or 118, wherein the pladienolide D or derivative comprises D12.

816. The antibody-drug conjugate of claim 125 or 126, wherein the arylpladienolide comprises D15.

817. The splicing regulator is D4: 【Chemistry 37】 The antibody-drug conjugate of any one of claims 1 to 106, comprising:

818. The antibody-drug conjugate of claim 817, wherein the splicing modulator comprises D4 and the linker comprises MC-Val-Cit-pABC.

819. The antibody-drug conjugate of claim 817, wherein the splicing modulator comprises D4 and the linker comprises MC-β-glucuronide.

820. The splicing regulator is D12: 【Transformation 38】 The antibody-drug conjugate of any one of claims 1 to 106, comprising:

821. The antibody-drug conjugate of claim 820, wherein the splicing modulator comprises D12 and the linker comprises MC-Val-Cit-pABC.

822. The antibody-drug conjugate of claim 820, wherein the splicing modulator comprises D12 and the linker comprises MC-β-glucuronide.

823. The splicing regulator is D15: 【Chemistry 39】 The antibody-drug conjugate of any one of claims 1 to 106, comprising:

824. The antibody-drug conjugate of claim 823, wherein the splicing modulator comprises D15 and the linker comprises MC-Val-Ala-pAB.

825. The antibody-drug conjugate of claim 137, wherein the antibody or antigen-binding fragment targets cells expressing HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, and / or STEAP1.

826. The antibody-drug conjugate of claim 156, wherein the antibody or antigen-binding fragment targets cells expressing HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFClB, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, and / or STEAP1.

827. Formula (I): Ab-(L-D) ｐ (I) (In the formula, Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is D4; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

828. The antibody-drug conjugate of claim 827, wherein the antibody or antigen-binding fragment targets HER2-expressing cells.

829. The antibody-drug conjugate of claim 828, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

830. The antibody-drug conjugate of claim 828 or 829, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3).

831. The antibody-drug conjugate of any one of claims 828 to 830, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

832. The antibody-drug conjugate of any one of claims 828 to 831, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

833. The antibody-drug conjugate of any one of claims 828 to 832, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

834. The antibody-drug conjugate of claim 827, wherein the antibody or antigen-binding fragment targets cells expressing HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, and / or STEAP1.

835. Formula (I): Ab-(L-D) ｐ (I) (In the formula, Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is D12; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

836. The antibody-drug conjugate of claim 835, wherein the antibody or antigen-binding fragment targets HER2-expressing cells.

837. The antibody-drug conjugate of claim 836, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

838. The antibody-drug conjugate of claim 836 or 837, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3).

839. The antibody-drug conjugate of any one of claims 836 to 838, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

840. The antibody-drug conjugate of any one of claims 836 to 839, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

841. The antibody-drug conjugate of any one of claims 836 to 840, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

842. The antibody-drug conjugate of claim 835, wherein the antibody or antigen-binding fragment targets cells expressing HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFClB, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, and / or STEAP1.

843. Formula (I): Ab-(L-D) ｐ (I) (In the formula, Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is D15; L is a linker that covalently links Ab to D; and p is an integer from 1 to 15. Antibody-drug conjugates of

844. The antibody-drug conjugate of claim 843, wherein the antibody or antigen-binding fragment targets HER2-expressing cells.

845. The antibody-drug conjugate of claim 844, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

846. The antibody-drug conjugate of claim 844 or 845, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3).

847. The antibody-drug conjugate of any one of claims 844 to 846, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

848. The antibody-drug conjugate of any one of claims 844 to 847, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

849. The antibody-drug conjugate of any one of claims 844 to 848, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

850. The antibody-drug conjugate of claim 843, wherein the antibody or antigen-binding fragment targets cells expressing HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFClB, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, and / or STEAP1.

851. The antibody-drug conjugate of any one of claims 827 to 850, wherein the linker is a cleavable linker.

852. The antibody-drug conjugate of claim 851, wherein the cleavable linker comprises MC-Val-Cit-pABC.

853. The antibody-drug conjugate of claim 851, wherein said cleavable linker comprises MC-Val-Ala-pABC.

854. The antibody-drug conjugate of claim 851, wherein the cleavable linker comprises MC-Val-Ala-pAB.

855. The antibody-drug conjugate of claim 851, wherein said cleavable linker comprises MC-Glu-Val-Cit-pABC.

856. The antibody-drug conjugate of claim 851, wherein said cleavable linker comprises MC-Ala-Ala-Asn-pABC.

857. The antibody-drug conjugate of claim 851, wherein the cleavable linker comprises MC-β-glucuronide.

858. The antibody-drug conjugate of any one of claims 827 to 850, wherein the linker is a non-cleavable linker.

859. The antibody-drug conjugate of claim 858, wherein the non-cleavable linker comprises Mal-Hex.

860. The antibody-drug conjugate of claim 858, wherein the non-cleavable linker comprises Mal-Et.

861. The antibody-drug conjugate of claim 858, wherein the non-cleavable linker comprises Mal-Et-O-Et.

862. The antibody-drug conjugate of any one of claims 827 to 861, wherein p is 1 to 10.

863. The antibody-drug conjugate of any one of claims 827 to 862, wherein p is 2 to 8.

864. The antibody-drug conjugate of any one of claims 827 to 863, wherein p is 4 to 8.

865. The antibody-drug conjugate of any one of claims 827 to 864, wherein p is 4.

866. The antibody-drug conjugate of any one of claims 827 to 864, wherein p is 8.

867. The antibody-drug conjugate of claim 181, wherein the cleavable linker comprises MC-Glu-Val-Cit-pABC.

868. 182. The antibody-drug conjugate of claim 181, wherein the cleavable linker comprises MC-Ala-Ala-Asn-pABC.

869. 9. A composition comprising a plurality of copies of the antibody-drug conjugate of any one of claims 776 to 868, wherein the average p of said antibody-drug conjugates in said composition is from about 3.5 to about 5.

5.

870. 9. A composition comprising a plurality of copies of the antibody-drug conjugate of any one of claims 776 to 868, wherein the average p of said antibody-drug conjugates in said composition is from about 7 to about 9.

871. 869 or 870. A method of treating a subject having or suspected of having a neoplastic disorder, comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 776-868 or the composition of claim 869 or 870.

872. 10. A method of reducing or inhibiting tumor growth in a subject having or suspected of having a neoplastic disorder, the method comprising administering to the subject a therapeutically effective amount of the antibody-drug conjugate of any one of claims 776-868 or the composition of claim 869 or 870.

873. 869 or 870, comprising providing a biological sample from the subject; and contacting the biological sample with the antibody-drug conjugate of any one of claims 776 to 868 or the composition of claim 869 or 870.

874. 871. The antibody-drug conjugate of any one of claims 776 to 868 or the composition of claim 869 or 870 for use in the treatment of a neoplastic disorder.

875. 870. Use of an antibody-drug conjugate according to any one of claims 776 to 868 or a composition according to claim 869 or 870 in the treatment of a neoplastic disorder.

876. 870。 Use of an antibody-drug conjugate of any one of claims 776 to 868 or a composition of claim 869 or 870 in a method for the manufacture of a medicament for the treatment of a neoplastic disorder.

877. 870. A pharmaceutical composition comprising the antibody-drug conjugate of any one of claims 776 to 868 or the composition of claim 869 or 870, and a pharmaceutically acceptable carrier.

878. A method for making an antibody-drug conjugate of any one of claims 776 to 868, comprising reacting an antibody or antigen-binding fragment with a linker attached to a splicing modulator under conditions that allow conjugation.

879. The method of any one of claims 303 to 687, wherein the antibody-drug conjugate comprises an antibody-drug conjugate of any one of claims 776 to 868.

880. The method of claim 688, wherein the antibody or antigen-binding fragment targets cells expressing HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFClB, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, and / or STEAP1.

881. 702. The method of claim 701, wherein said cleavable linker comprises MC-Glu-Val-Cit-pABC.

882. 702. The method of claim 701, wherein said cleavable linker comprises MC-Ala-Ala-Asn-pABC.

883. 702. The method of claim 701, wherein the cleavable linker comprises MC-β-glucuronide.

884. The splicing regulator is D4: 【Chemistry 40】 The method of any one of claims 688 to 708, comprising:

885. The method of claim 884, wherein the splicing modulator comprises D4 and the linker comprises MC-Val-Cit-pABC.

886. The method of claim 884, wherein the splicing modulator comprises D4 and the linker comprises MC-β-glucuronide.

887. The splicing regulator is D12: 【Chemistry 41】 The method of any one of claims 688 to 708, comprising:

888. The method of claim 887, wherein the splicing modulator comprises D12 and the linker comprises MC-Val-Cit-pABC.

889. The method of claim 887, wherein the splicing modulator comprises D12 and the linker comprises MC-β-glucuronide.

890. The splicing regulator is D15: 【Chemistry 42】 The method of any one of claims 688 to 708, comprising:

891. The method of claim 890, wherein the splicing modulator comprises D15 and the linker comprises MC-Val-Ala-pAB.

892. The method of claim 719 or 720, wherein the pladienolide D or derivative comprises D4.

893. The method of claim 719 or 720, wherein the pladienolide D or derivative comprises D12.

894. The method of claim 727 or 728, wherein the arylpladienolide comprises D15.

895. 10. A neo-antigen vaccine comprising at least one neo-antigen peptide, wherein said at least one neo-antigen peptide comprises a modified or novel neo-antigen sequence induced by contacting a neoplastic cell with an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 776-868, or a composition of claim 869 or 870.

896. 10. A neo-antigen vaccine comprising at least one neo-antigen mRNA, wherein said at least one neo-antigen mRNA encodes a modified or novel neo-antigen sequence induced by contacting a neoplastic cell with an effective amount of a splicing modulator, an antibody-drug conjugate of any one of claims 776-868, or a composition of claim 869 or 870.

897. structure 【Chemistry 43】 (wherein Ab is an antibody or antigen-binding fragment covalently bound to the maleimide group of ADL1-D1 through the sulfur atom of a thiol group on the antibody or antigen-binding fragment). An antibody-drug conjugate comprising:

898. structure 【Chemistry 44】 (wherein Ab is an antibody or antigen-binding fragment covalently bound to the maleimide group of ADL1-D4 through the sulfur atom of a thiol group on the antibody or antigen-binding fragment). An antibody-drug conjugate comprising:

899. structure 【Chemistry 45】 wherein Ab is an antibody or antigen-binding fragment covalently bound to the maleimide group of ADL1-D12 through the sulfur atom of a thiol group on the antibody or antigen-binding fragment. An antibody-drug conjugate comprising:

900. structure 【Chemistry 46】 wherein Ab is an antibody or antigen-binding fragment covalently bound to the maleimide group of ADL5-D2 through the sulfur atom of a thiol group on the antibody or antigen-binding fragment. An antibody-drug conjugate comprising:

901. structure 【Chemistry 47】 where Ab is an antibody or antigen-binding fragment covalently bound to the maleimide group of ADL5-D15 through the sulfur atom of a thiol group on the antibody or antigen-binding fragment. An antibody-drug conjugate comprising:

902. structure 【Chemistry 48】 wherein Ab is an antibody or antigen-binding fragment covalently bound to the maleimide group of ADL13-D4 through the sulfur atom of a thiol group on the antibody or antigen-binding fragment. An antibody-drug conjugate comprising:

903. The antibody-drug conjugate of any one of claims 897 to 902, wherein the antibody or antigen-binding fragment targets a HER2-expressing cell.

904. The antibody-drug conjugate of claim 903, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

905. The antibody-drug conjugate of claim 903 or 904, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) comprising the amino acid sequences of SEQ ID NO: 1 (HCDR1), SEQ ID NO: 2 (HCDR2), and SEQ ID NO: 3 (HCDR3); and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) comprising the amino acid sequences of SEQ ID NO: 4 (LCDR1), SEQ ID NO: 5 (LCDR2), and SEQ ID NO: 6 (LCDR3).

906. The antibody-drug conjugate of any one of claims 903 to 905, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:

20.

907. The antibody-drug conjugate of any one of claims 903 to 906, wherein the antibody or antigen-binding fragment comprises a human IgG1 heavy chain constant region.

908. The antibody-drug conjugate of any one of claims 903 to 907, wherein the antibody or antigen-binding fragment comprises a human Ig kappa light chain constant region.

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