Splicing modulating antibody-drug conjugates and methods of use thereof

Abstract

The invention relates to splicing modulating antibody-drug conjugates and methods of using the same. Linker-drug compounds and antibody-drug conjugates that bind to human tumor targets are disclosed. These linker-drug compounds and antibody-drug conjugates comprise a splicing modulator drug moiety. The disclosure further relates to methods and compositions for treating neoplastic disorders by administering the antibody-drug conjugates provided herein. In some embodiments, the splicing modifier comprises a piradienolide or a piradienolide derivative.

Description

[0001] This application is a divisional application of Chinese invention patent application No. 201980036786.3 (PCT application number PCT / US2019 / 035015), filed on May 31, 2019, entitled "Splice-regulating antibody-drug conjugate and method of use thereof".

[0002] This disclosure claims priority to the following: 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 which are incorporated herein by reference in their entirety.

[0003] This 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. This disclosure further relates to methods and compositions that can be used to treat or diagnose cancer, and methods for preparing these compositions, wherein the cancer expresses a target antigen and / or is suitable for treatment by disrupting RNA splicing.

[0004] Most protein-coding genes in the human genome consist of multiple exons (coding regions) separated by introns (non-coding regions). Gene expression produces a single precursor messenger RNA (pre-mRNA). Subsequently, intron sequences are removed from the pre-mRNA through a process called splicing, producing mature messenger RNA (mRNA). By including different combinations of exons, alternative splicing can produce mRNAs encoding different protein variants.

[0005] RNA splicing is catalyzed by the spliceosome, a dynamic multi-protein-RNA complex composed of five small nuclear RNAs (snRNAs U1, U2, U4, U5, and U6) and associated proteins. This spliceosome assembles on pre-mRNA to establish a dynamic cascade of multiple RNA and protein interactions that catalyze intron excision and exon joining (Matera and Wang (2014) Nat Rev Mol Cell Biol. [Nature Reviews Molecular and Cell Biology] 15(2):108-21). Increasing evidence links human diseases to dysregulation of RNA splicing affecting many genes (Scotti and Swanson (2016) Nat Rev Genet. [Nature Reviews Genetics] 17(1):19-32).

[0006] The spliceosome is an important target in cancer biology. Several studies have demonstrated significant changes in the splicing profile of cancer cells and the splicing factors themselves (Agrawal et al. (2018) Curr Opin Genet Dev. [New Insights in Genetics and Development] 48:67-74). Alternative splicing can lead to different exon inclusion / exclusion, intron retention, or the use of cryptic splicing sites (Seiler et al. (2018) Cell Rep. [Cell Reports] 23(1):282-296). In summary, these events cause functional changes that can lead to tumorigenesis or resistance to treatment (Siegfried and Karni (2018) Curr Opin Genet Dev. [New Insights in Genetics and Development] 48:16-21).

[0007] Certain natural products can bind to the SF3b spliceosome complex. These small molecules regulate splicing by promoting intron retention and / or exon skipping (Teng et al. (2017) Nat Commun. [Nature Communications] 8:15522). A significant portion of the resulting transcripts contain premature stop codons, triggering nonsense-mediated mRNA decay (NMD). Furthermore, due to impaired typical splicing, typical transcripts are drastically reduced, which can adversely affect cell function and viability. For this reason, splicing regulators have become a promising class of drugs for cancer treatment (Puthenveetil et al. (2016) Bioconjugate Chem. [Bioconjugate Chemistry] 27:1880-8).

[0008] The proto-oncogene human epidermal growth factor receptor 2 (HER2) encodes a transmembrane tyrosine kinase receptor that belongs to the human epidermal growth factor receptor (EGFR) family (King et al. (1985) Science 229:974-6). Overexpression of HER2 can constitutively activate growth factor signaling pathways, such as the PI3K-AKT-mTOR pathway, and thus act as an oncogenic driver in several cancer types, including approximately 20% of invasive breast cancer (Slamon et al. (1989) Science 244:707-12; Gajria and Chandarlapaty (2011) Expert Rev Anticancer Ther. 11:263-75). Given the phenotype of HER2 amplification-mediated transformation and the fact that HER2 expression is primarily confined 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 delivery of cancer therapies include, but are not limited to, CD138 (also known as multiligand glycan-1) and hepatic ligand A receptor 2 (EPHA2).

[0009] CD138 is a cell surface heparan sulfate proteoglycan that is essential for maintaining cell morphology and interacting with the surrounding microenvironment (Akl et al. (2015) Oncotarget 6(30):28693-715; Szatmári et al. (2015) Dis Markers 2015:796052). Typically, loss of CD138 expression in cancer cells reduces cell adhesion to the extracellular matrix and enhances cell motility and invasiveness (Teng et al. (2012) MatrixBiol. 31:3-16). Increased matrix CD138 expression also alters fibronectin production and extracellular matrix organization (Yang et al. (2011) Am J Pathol. 178:325-35). Furthermore, increased CD138 expression in 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 malignant tumor of plasma cells). Therefore, CD138 is a compelling antigen for targeted therapy of several cancers and other hematologic malignancies (Sherbenou et al. (2015) Blood Rev. 29(2):81-91; Wijdenes et al. (1996) Br J Haematol. 94(2):318-23).

[0010] EPHA2 is a transmembrane glycoprotein that is heavily overexpressed in several malignant cancer-derived cell lines and advanced cancer forms (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). EPHA2 protein is highly overexpressed in terms of both the percentage of tumor cells and the percentage of cells within the tumor in patients, and it is a plasma membrane-localized receptor that can be internalized upon ligand binding (Walker-Daniels et al. (2002) Mol Cancer Res. [Molecular Cancer Research] 1:79-87). Furthermore, EPHA2 expression is associated with poor prognosis, increased metastasis, and decreased survival. Therefore, EPHA2, due to its expression patterns, localization, and functional importance in cancer patient outcomes, represents another compelling antigen for the targeted delivery of novel anticancer therapies.

[0011] In various embodiments, this disclosure provides, in part, novel compounds that are biologically active against neoplastic cells. These compounds can slow, inhibit, and / or reverse tumor growth in mammals and may be used to treat human cancer patients. In various embodiments, this disclosure provides novel antibody-drug conjugates employing these novel compounds or other functional splicing inhibitor molecules.

[0012] More specifically, in various embodiments, this disclosure relates to antibody-drug conjugate (ADC) complexes capable of binding to and killing proliferation cells. In various embodiments, the ADC complexes disclosed herein include a linker that connects a splicing regulator to a full-length antibody or antigen-binding fragment. In various embodiments, these ADC complexes are also capable of internalization into target cells after binding.

[0013] In different embodiments, the ADC complex can be represented by formula (I):

[0014] Ab-(LD) p (I)

[0015] Ab is an antibody or its antigen-binding fragment that targets neoplastic cells or another tumor-associated target;

[0016] D is a splice modifier;

[0017] L is the connector that covalently links Ab to D; and

[0018] p is an integer from 1 to 15.

[0019] In different embodiments, the ADC complex can be represented by formula (I):

[0020] Ab-(LD) p (I)

[0021] Ab is an antibody or its antigen-binding fragment that targets neoplastic cells;

[0022] D is a splice modifier with formula (II):

[0023] or a pharmaceutically acceptable salt thereof, wherein:

[0024] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0025] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[0026] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0027] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups and -NR 15 R 16 ;

[0028] R 15 and R 16Each is independently selected from hydrogen, R 17 -C(=O)-R 17 and -C(=O)-OR 17 ;

[0029] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0030] Z is selected from

[0031] Where R 1 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups.

[0032] Where R 6 and R 7 At least one of them is hydrogen;

[0033] And L is the connector that covalently connects Ab to D; and

[0034] p is an integer from 1 to 15.

[0035] In some embodiments, the antibody or antigen-binding fragment is capable of internalization into target cells. In some embodiments, the linker is covalently linked to a splicing regulator (“LD”) of formula (II), and the LD has the structure of formula (II-A):

[0036] Or its pharmaceutically acceptable salt.

[0037] Z' is selected from and

[0038] All other variables are defined as per equation (II).

[0039] In various other embodiments, the ADC complex may be represented by formula (I):

[0040] Ab-(LD) p (I)

[0041] Ab is an antibody or its antigen-binding fragment that targets neoplastic cells;

[0042] D is a splice modifier with formula (IV):

[0043] or a pharmaceutically acceptable salt thereof, wherein:

[0044] R 1 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, -OC(=O)-(C1-C6 alkyl) groups, and -CD3;

[0045] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[0046] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0047] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups and -NR 15 R 16 ;

[0048] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and -C(=O)-OR 17 ;and

[0049] R 17Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0050] Where R 1 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups.

[0051] Where R 6 and R 7 At least one of them is hydrogen;

[0052] And L is the connector that covalently connects Ab to D; and

[0053] p is an integer from 1 to 15.

[0054] In some embodiments, the antibody or antigen-binding fragment is capable of internalization into target cells. In some embodiments, the linker is covalently linked to a splicing regulator (“LD”), and the LD has a structure of formula (IV-A):

[0055] Or its pharmaceutically acceptable salt.

[0056] In various other embodiments, the ADC complex may be represented by formula (I):

[0057] Ab-(LD) p (I)

[0058] Ab is an antibody or its antigen-binding fragment that targets neoplastic cells;

[0059] D is a splice modifier with formula (VI):

[0060] or a pharmaceutically acceptable salt thereof, wherein:

[0061] R 1 and R 9Each group is independently selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, -OC(=O)-(C1-C6 alkyl) groups, and -CD3;

[0062] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[0063] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0064] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[0065] R 10 Selected from hydrogen, C1-C6 alkyl groups, -C(=O)-(C1-C6 alkyl) groups, and -CD3;

[0066] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and

[0067] -C(=O)-OR 17 ;

[0068] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0069] a is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0070] where R 1 R3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[0071] where R 6 and R 7 At least one of them is hydrogen; and

[0072] where R 1 and R 9 They cannot both be absent at the same time;

[0073] And L is the connector that covalently connects Ab to D; and

[0074] p is an integer from 1 to 15.

[0075] In some embodiments, the antibody or antigen-binding fragment is capable of internalization into target cells. In some embodiments, the linker is covalently linked to a splicing regulator (“LD”), and the LD has a structure of formula (VI-A):

[0076] Or its pharmaceutically acceptable salt.

[0077] In various other embodiments, the ADC complex may be represented by formula (I):

[0078] Ab-(LD) p (I)

[0079] Ab is an antibody or its antigen-binding fragment that targets neoplastic cells;

[0080] D is a splice modifier with formula (VIII):

[0081] or a pharmaceutically acceptable salt thereof, wherein:

[0082] R 1Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0083] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[0084] R 4 Selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups; and

[0085] R 10 Selected from 3- to 10-membered carbon rings and 3- to 10-membered heterocycles, each of which is composed of 0 to 3 R... a Replace, where each R a Independently selected from halogens, C1-C6 alkyl groups, -O-(C1-C6)alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylhydroxy groups, and -S (=O). w -(4- to 7-membered heterocycles), 4- to 7-membered carbon rings and 4- to 7-membered heterocycles;

[0086] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and -C(=O)-OR 17 ;and

[0087] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0088] where R 1 R 3 R 4 R 10 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0089] Each R a Independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, -NR 15 R 16 C1-C6 alkyl groups, -(C=O)-(C1-C6 alkyl) groups, -(C=O)-(C1-C6 alkyl)-(C3-C 10 Heterocyclic groups), -S (=O) w -(C3-C8 heterocyclic) groups and C1-C6 alkylcarboxylic acid groups, each substituted by 0, 1, or 2 independently selected groups from: halogens, hydroxyl groups, -NR groups. 15 R 16 and C1-C3 alkyl groups; and

[0090] w is 0, 1, or 2;

[0091] And L is the connector that covalently connects Ab to D; and

[0092] p is an integer from 1 to 15.

[0093] In some embodiments, the antibody or antigen-binding fragment is capable of internalization into target cells. In some embodiments, the linker is covalently linked to a splicing regulator (“LD”), and the LD has a structure of formula (VIII-A):

[0094] Or its pharmaceutically acceptable salt.

[0095] In some embodiments, the splice modifier comprises a modifier of the SF3b complex.

[0096] In some embodiments, the splice modifier comprises pladienolide or a pladienolide derivative. In some embodiments, the splice modifier comprises pladienolide D or a pladienolide D derivative. In some embodiments, the pladienolide D or its derivative comprises D2, D1, D4, D8, D10, D11 (E7107), D20, D21, D22, D12, or D25. In some embodiments, the pladienolide D or its derivative comprises D2. In some embodiments, the pladienolide D or its derivative comprises D1. In some embodiments, the pladienolide D or its derivative comprises D4. In some embodiments, the pladienolide D or its derivative comprises D12.

[0097] In some embodiments, the prasadiene lactone D or its derivative is a zwitterionic prasadiene lactone D or its derivative. In some embodiments, the zwitterionic prasadiene lactone D or its derivative comprises D22 or D25.

[0098] In some other embodiments, the splice modulator comprises prasadiene lactone B or a derivative thereof. In some embodiments, the prasadiene lactone B or its derivative comprises D9, D18, D19, or D13.

[0099] In some embodiments, the splice modifier comprises arylpradoadiene lactone. In some embodiments, the arylpradoadiene lactone comprises D15, D14, D16, D17, D26, or D33. In some embodiments, the arylpradoadiene lactone comprises D15. In some embodiments, the arylpradoadiene lactone is a zwitterionic arylpradoadiene lactone. In some embodiments, the zwitterionic arylpradoadiene lactone comprises D33.

[0100] In some embodiments, the splice modulator comprises D1:

[0101]

[0102] In some embodiments, the splice modulator comprises D2:

[0103]

[0104] In some embodiments, the splice modulator comprises D3:

[0105]

[0106] In some embodiments, the splice modulator comprises D4:

[0107]

[0108] In some embodiments, the splice modulator comprises D4':

[0109]

[0110] In some embodiments, the splice modulator comprises D5:

[0111]

[0112] In some embodiments, the splice modulator comprises D6:

[0113]

[0114] In some embodiments, the splice modulator comprises D7:

[0115]

[0116] In some embodiments, the splice modulator comprises D8:

[0117]

[0118] In some embodiments, the splice modulator comprises D9:

[0119]

[0120] In some embodiments, the splice modulator comprises D10:

[0121]

[0122] In some embodiments, the splice modulator comprises D11:

[0123]

[0124] In some embodiments, the splice modulator comprises D12:

[0125]

[0126] In some embodiments, the splice modulator comprises D13:

[0127]

[0128] In some embodiments, the splice modulator comprises D14:

[0129]

[0130] In some embodiments, the splice modulator comprises D15:

[0131]

[0132] In some embodiments, the splice modulator comprises D16:

[0133]

[0134] In some embodiments, the splice modulator comprises D17:

[0135]

[0136] In some embodiments, the splice modulator comprises D18:

[0137]

[0138] In some embodiments, the splice modulator comprises D19:

[0139]

[0140] In some embodiments, the splice modulator comprises D20:

[0141]

[0142] In some embodiments, the splice modulator comprises D21:

[0143]

[0144] In some embodiments, the splice modulator comprises D22:

[0145]

[0146] In some embodiments, the splice modulator comprises D23:

[0147]

[0148] In some embodiments, the splice modulator comprises D24:

[0149]

[0150] In some embodiments, the splice modulator comprises D25:

[0151]

[0152] In some embodiments, the splice modulator comprises D26:

[0153]

[0154] In some embodiments, the splice modulator comprises D27:

[0155]

[0156] In some embodiments, the splice modulator comprises D28:

[0157]

[0158] In some embodiments, the splice modulator comprises D29:

[0159]

[0160] In some embodiments, the splice modulator comprises D30:

[0161]

[0162] In some embodiments, the splice modulator comprises D31:

[0163]

[0164] In some embodiments, the splice modulator comprises D32:

[0165]

[0166] In some embodiments, the splice modulator comprises D33:

[0167]

[0168] In some embodiments, the splice modulator comprises D34:

[0169]

[0170] In some embodiments, the splice modulator comprises D35:

[0171]

[0172] In some embodiments, the splice modulator comprises one of the pharmaceutical portions listed in Table 7. In some embodiments, the splice 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.

[0173] In some embodiments, a splice modulator is disclosed, and its use as a therapeutic agent, either alone or as part of an ADC. In some embodiments, the splice modulator comprises D4, D4', D12, D15, D8, D9, D10, D13, D18, D19, D20, D21, D22, D25, or D33.

[0174] In some embodiments, the splice modulator comprises D4 and the connector comprises MC-Val-Cit-pABC. In some embodiments, the splice modulator comprises D4 and the connector comprises MC-β-glucuronide. In some embodiments, the splice modulator comprises D12 and the connector comprises MC-Val-Cit-pABC. In some embodiments, the splice modulator comprises D12 and the connector comprises MC-β-glucuronide. In some embodiments, the splice modulator comprises D15 and the connector comprises MC-Val-Ala-pAB.

[0175] In various embodiments, the linkers used in the ADCs disclosed herein are stable outside the cell, such that the ADC remains intact when present in extracellular conditions but can be cleaved upon internalization into cells (e.g., tumor or cancer cells). In some embodiments, the splicing regulator cleaves from the antibody or antigen-binding fragment when the ADC enters a cell (the cell expresses an antigen targeted by an antibody or antigen-binding fragment in the ADC). In some embodiments, the linker is a cleavable linker.

[0176] In some embodiments, the linker comprises a cleavable peptide moiety. In some embodiments, the cleavable peptide moiety is enzymatically cleavable. 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”).

[0177] In some embodiments, the connector includes a cleavable glucuronide moiety. In some embodiments, the cleavable glucuronide moiety can be cleaved by an enzyme. In some embodiments, the cleavable glucuronide moiety can be cleaved by a glucuronidase. In some embodiments, the cleavable glucuronide moiety can be cleaved by a β-glucuronidase.

[0178] In some embodiments, the connector includes at least one spacer subunit. In some embodiments, the spacer subunit or connector includes a polyethylene glycol (PEG) portion. In some embodiments, the PEG portion includes -(PEG). m - And m is an integer from 1 to 10. In some embodiments, m is 2. In some other embodiments, the spacer subunit or connector comprises an alkyl portion. In some embodiments, the alkyl portion comprises -(CH2). n - And 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.

[0179] In some embodiments, the spacer subunit is linked to the antibody or antigen-binding fragment via a maleimide (Mal) portion (“Mal-spacer subunit”). In some embodiments, the Mal-spacer subunit may react with cysteine ​​residues on the antibody or antigen-binding fragment. In some embodiments, the Mal-spacer subunit is attached to the antibody or antigen-binding fragment via cysteine ​​residues on the antibody or antigen-binding fragment.

[0180] In some embodiments, the connector includes the Mal-spacer subunit and a cleavable peptide portion. In some embodiments, the cleavable peptide portion includes an amino acid unit. In some embodiments, the cleavable peptide portion or amino acid unit includes Val-Cit. In some embodiments, the cleavable peptide portion or amino acid unit includes Val-Ala. In some embodiments, the cleavable peptide portion or amino acid unit includes Glu-Val-Cit. In some embodiments, the cleavable peptide portion or amino acid unit includes Ala-Ala-Asn. In some embodiments, the Mal-spacer subunit includes an alkyl portion. In some embodiments, the Mal-spacer subunit includes a PEG portion. In some embodiments, the Mal-spacer subunit includes maleimide hexanoyl (MC).

[0181] In some embodiments, the Mal-spacer subunit links the antibody or antigen-binding fragment to a cleavable portion in the adapter. In some embodiments, the cleavable portion in the adapter comprises a cleavable peptide portion. In some embodiments, the cleavable peptide portion comprises an amino acid unit. In some embodiments, the cleavable peptide portion or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn. In some embodiments, the adapter comprises MC-Val-Cit. In some embodiments, the adapter comprises MC-Val-Ala. In some embodiments, the adapter comprises MC-Glu-Val-Cit. In some embodiments, the adapter comprises MC-Ala-Ala-Asn. In some embodiments, the Mal-spacer subunit comprises an alkyl portion. In some embodiments, the Mal-spacer subunit comprises a PEG portion. In some embodiments, the Mal-spacer subunit comprises maleimide hexanoyl (MC).

[0182] In some embodiments, the cleavable portion of the connector directly binds to the splicing modulator, or a spacer unit connects the cleavable portion of the connector to the splicing modulator. In some embodiments, cleavage of the conjugate releases the splicing modulator from the antibody or antigen-binding fragment and the connector. In some embodiments, the spacer unit that connects the cleavable portion of the connector to the splicing modulator is self-immolative.

[0183] In some embodiments, the spacer subunit connecting the cleavable portion of the connector to the splicing regulator comprises a p-aminobenzyloxycarbonyl group (pABC). In some embodiments, the pABC connects the cleavable portion of the connector to the splicing regulator. In some embodiments, the cleavable portion of the connector comprises a cleavable peptide portion. In some embodiments, the cleavable peptide portion comprises an amino acid unit. In some embodiments, the cleavable peptide portion or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn. In some embodiments, the connector comprises Val-Cit-pABC. In some other embodiments, the connector comprises Val-Ala-pABC. In some embodiments, the connector comprises Glu-Val-Cit-pABC. In some embodiments, the connector comprises Ala-Ala-Asn-pABC.

[0184] In some embodiments, the spacer subunit connecting the cleavable portion of the connector to the splicing regulator comprises p-aminobenzyl (pAB). In some embodiments, pAB connects the cleavable portion of the connector to the splicing regulator. In some embodiments, the cleavable portion of the connector comprises a cleavable peptide portion. In some embodiments, the cleavable peptide portion comprises an amino acid unit. In some embodiments, the cleavable peptide portion or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn. In some embodiments, the connector comprises Val-Cit-pAB. In some other embodiments, the connector comprises Val-Ala-pAB. In some other embodiments, the connector comprises Glu-Val-Cit-pAB. In some other embodiments, the connector comprises Ala-Ala-Asn-pAB.

[0185] In various embodiments, the linker is a non-cleavable linker. In some embodiments, the splicing regulator in the ADC is released through the degradation of the antibody or antigen-binding fragment. In some embodiments, the linker maintains covalent association with at least one amino acid of the antibody and the drug when internalized and degraded within the target cells.

[0186] In some embodiments, the connector is a non-disintegrable connector comprising at least one spacer subunit. In some embodiments, the spacer subunit or connector comprises a polyethylene glycol (PEG) portion. In some embodiments, the PEG portion comprises -(PEG). m - And m is an integer from 1 to 10. In some embodiments, m is 2. In some other embodiments, the spacer subunit or connector comprises an alkyl portion. In some embodiments, the alkyl portion comprises -(CH2).n -or-(CH2) n -O-(CH2) n And 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.

[0187] In some embodiments, the spacer subunit in the cleavable linker is linked to the antibody or antigen-binding fragment via a maleimide (Mal) portion (“Mal-spacer subunit”). In some embodiments, the Mal-spacer subunit may react with cysteine ​​residues on the antibody or antigen-binding fragment. In some embodiments, the Mal-spacer subunit is attached to the antibody or antigen-binding fragment via cysteine ​​residues on the antibody or antigen-binding fragment. In some embodiments, the Mal-spacer subunit comprises an alkyl portion. In some embodiments, the Mal-spacer subunit comprises a PEG portion. In some embodiments, the linker or Mal-spacer subunit comprises maleimide hexanoyl (MC). In some embodiments, the linker or Mal-spacer subunit comprises maleimide hexanoyl (MC) and at least one additional spacer subunit. In some embodiments, the linker or Mal-spacer subunit comprises MC-(PEG)2. In some embodiments, the linker or Mal-spacer subunit comprises MC-(PEG)2 and at least one additional spacer subunit. In some embodiments, the linker or Mal-spacer subunit comprises Mal-Hex. In some embodiments, the connector or Mal-spacer subunit comprises Mal-Hex and at least one additional spacer subunit. In some embodiments, the connector or Mal-spacer subunit comprises Mal-Et. In some embodiments, the connector or Mal-spacer subunit comprises Mal-Et and at least one additional spacer subunit. In some embodiments, the connector or Mal-spacer subunit comprises Mal-Et-O-Et. In some embodiments, the connector or Mal-spacer subunit comprises Mal-Et-O-Et and at least one additional spacer subunit. In some embodiments, the Mal-spacer subunit links the antibody or antigen-binding fragment to the splicing modulator.

[0188] In different embodiments, the ADC complex can be represented by formula (I):

[0189] Ab-(LD) p (I)

[0190] Where Ab is an antibody or its antigen-binding fragment (e.g., any of the antibody or binding domain sequences disclosed herein) that targets neoplastic cells or another tumor-associated target (e.g., cancer antigen); D is any small molecule suitable for treating cancer (e.g., a splicing regulator, such as any of the splicing regulators disclosed herein); L is a linker that covalently links Ab to D (e.g., any of the linkers disclosed herein); and p is an integer from 1 to 15.

[0191] In some embodiments, Ab is selected from any of the antibody or binding domain sequences disclosed herein. In some embodiments, Ab is an antibody or binding domain sequence targeting HER2 and / or HER2-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting CD138 and / or CD138-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting EPHA2 and / or EPHA2-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting MSLN and / or MSLN-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting FOLH1 and / or FOLH1-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting CDH6 and / or CDH6-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting CEACAM5 and / or CEACAM5-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting CFC1B and / or CFC1B-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting ENPP3 and / or ENPP3-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting FOLR1 and / or FOLR1-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting HAVCR1 and / or HAVCR1-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting KIT and / or KIT-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting MET and / or MET-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting MUC16 and / or MUC16-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting SLC39A6 and / or SLC39A6-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence targeting SLC44A4 and / or SLC44A4-expressing cytoplasmic cells. In some embodiments, Ab is an antibody or binding domain sequence that targets STEAP1 and / or STEAP1-expressing proliferative cells. In some embodiments, Ab is an antibody or binding domain sequence that targets another cancer antigen.

[0192] In some embodiments, D is a splice modulator. In some embodiments, D is selected from any of the splice modulators disclosed herein. In some embodiments, D is selected from the splice modulators 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 selected from the splice modulators 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 splice modulator comprising D2 or any derivative thereof. In some embodiments, D is a splice modulator comprising D1 or any derivative thereof.

[0193] In some embodiments, L is selected from any of the connectors disclosed herein, or any combination of the connector components disclosed herein. In some embodiments, L is a connector 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 connector may also include one or more additional spacer subunits. In some embodiments, L is a connector ADL1, ADL2, ADL5, ADL6, ADL7, ADL10, ADL12, ADL13, ADL14, ADL15, ADL21, ADL22, or ADL23. In some embodiments, L is a connector ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15. In some embodiments, L is an ADL12, ADL14, or ADL15 connector. In some embodiments, the ADL1, ADL2, ADL5, ADL6, ADL7, ADL12, ADL13, ADL14, ADL21, ADL23, or ADL15 connector may also include one or more additional spacer sub-units. In some embodiments, L is an ADL1 connector and may optionally include one or more additional spacer sub-units. In some embodiments, L is an ADL2 connector and may optionally include one or more additional spacer sub-units. In some embodiments, L is an ADL5 connector and may optionally include one or more additional spacer sub-units. In some embodiments, L is an ADL6 connector and may optionally include one or more additional spacer sub-units. In some embodiments, L is an ADL7 connector and may optionally include one or more additional spacer sub-units. In some embodiments, L is an ADL12 connector and may optionally include one or more additional spacer sub-units. In some embodiments, L is an ADL14 connector and may optionally include one or more additional spacer sub-units. In some embodiments, L is an ADL15 connector and may optionally include one or more additional spacer sub-units. In different embodiments of the ADC described herein, p is 1 to 10. In different embodiments, p is 2 to 8. In different embodiments, p is 4 to 8. In some embodiments, p is 4. In some embodiments, p is 8.

[0194] In some embodiments, the LD having formula (I) is ADL1-D1. In some embodiments, the LD having formula (I) is ADL6-D1. In some embodiments, the LD having formula (I) is ADL5-D2. In some embodiments, the LD having formula (I) is ADL1-D18. In some embodiments, the LD having formula (I) is ADL5-D19. In some embodiments, the LD having formula (I) is ADL14-D1. In some embodiments, the LD having formula (I) is ADL12-D1. In some embodiments, the LD having formula (I) is ADL15-D1. In some embodiments, the LD having formula (I) is ADL12-D20. In some embodiments, the LD having formula (I) is ADL10-D1. In some embodiments, the LD having formula (I) is ADL12-D2. In some embodiments, the LD having formula (I) is ADL15-D2. In some embodiments, the LD having formula (I) is ADL12-D21. In some embodiments, the LD having formula (I) is ADL6-D9. In some embodiments, the LD having formula (I) is ADL1-D4. In some embodiments, the LD having formula (I) is ADL1-D3. In some embodiments, the LD having formula (I) is ADL1-D12. In some embodiments, the LD having formula (I) is ADL1-D7. In some embodiments, the LD having formula (I) is ADL1-D6. In some embodiments, the LD having formula (I) is ADL1-D5. In some embodiments, the LD having formula (I) is ADL22-D4. In some embodiments, the LD having formula (I) is ADL5-D10. In some embodiments, the LD having formula (I) is ADL5-D11. In some embodiments, the LD having formula (I) is ADL1-D13. In some embodiments, the LD having formula (I) is ADL1-D8. In some embodiments, the LD having formula (I) is ADL1-D22. In some embodiments, the LD having formula (I) is ADL5-D25. In some embodiments, the LD having formula (I) is ADL12-D22. In some embodiments, the LD having formula (I) is ADL5-D15. In some embodiments, the LD having formula (I) is ADL1-D14. In some embodiments, the LD having formula (I) is ADL5-D26. In some embodiments, the LD having formula (I) is ADL1-D16. In some embodiments, the LD having formula (I) is ADL5-D17. In some embodiments, the LD having formula (I) is ADL1-D33. In some embodiments, the LD having formula (I) is ADL1-D28. In some embodiments, the LD having formula (I) is ADL1-D31.In some embodiments, the LD having formula (I) is ADL1-D29. In some embodiments, the LD having formula (I) is ADL1-D35. In some embodiments, the LD having formula (I) is ADL5-D32. In some embodiments, the LD having formula (I) is ADL5-D27. In some embodiments, the LD having formula (I) is ADL12-D35. In some embodiments, the LD having formula (I) is ADL12-D28. In some embodiments, the LD having formula (I) is ADL1-D23. In some embodiments, the LD having formula (I) is ADL1-D24.

[0195] In some embodiments, an ADC pool is provided, whereby random conjugation occurs, and the average p in the pool is between about 2 and about 8. In some embodiments, an ADC pool is provided, whereby random conjugation occurs, and the average p in the pool is between about 4 and about 8. In some embodiments, an ADC pool is provided, whereby random conjugation occurs, and the average p in the pool is about 4. In some embodiments, an ADC pool is provided, whereby random conjugation occurs, and the average p in the pool is about 8. This document provides compositions (e.g., pharmaceutical compositions) comprising multiple copies of any of the ADCs, wherein the average drug loading (average p) of the ADC in the composition is between about 3.5 and about 5.5 (e.g., about 4), or between about 7 and about 9 (e.g., about 8).

[0196] In some embodiments, the antibody or antigen-binding fragment (Ab) in the ADC targets cytoplasmic cells derived from hematologic malignancies or solid tumors. In some embodiments, the antibody or antigen-binding fragment targets cytoplasmic cells derived from hematologic malignancies. In some embodiments, the hematologic malignancy is selected from B-cell malignancies, leukemia (e.g., acute myeloid leukemia), lymphoma, and myeloma (e.g., multiple myeloma). In some embodiments, the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the antibody or antigen-binding fragment targets cytoplasmic cells derived from solid tumors. 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 gland duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney 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.

[0197] In various embodiments, the antibody or antigen-binding fragment (Ab) in the ADC is an anti-HER2 antibody or an antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment binds to HER2 and targets HER2-expressing neoplasms (i.e., the ADC targets HER2-expressing neoplasms). In some embodiments, the antibody or antigen-binding fragment in the ADC is an internalized anti-HER2 antibody or an internalized antigen-binding fragment thereof.

[0198] In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing 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 an internalizing antigen-binding fragment. In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises a human frame sequence. In some embodiments, the anti-HER2 antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing 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 Igκ or λ light chain constant region. In some embodiments, the anti-HER2 antibody or antigen-binding fragment competes with and / or binds to the same epitope as an antibody comprising the heavy chain variable domain of SEQ ID NO:19 and the light chain variable domain of SEQ ID NO:20.

[0199] In various embodiments, the antibody or antigen-binding fragment (Ab) in the ADC is an anti-CD138 antibody or an antigen-binding fragment thereof. In some embodiments, the antibody or antigen-binding fragment binds to CD138 and targets CD138-expressing neoplasms (i.e., the ADC targets CD138-expressing neoplasms). In some embodiments, the antibody or antigen-binding fragment in the ADC is an internalized anti-CD138 antibody or an internalized antigen-binding fragment thereof.

[0200] In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing 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 an internalizing antigen-binding fragment. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a human frame sequence. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:21 and a light chain variable region containing the amino acid sequence of SEQ ID NO:22. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a mouse IgG2a heavy chain constant region. In some embodiments, the anti-CD138 antibody or antigen-binding fragment comprises a mouse Igκ 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 Igκ or λ light chain constant region. In some embodiments, the anti-CD138 antibody or antigen-binding fragment competes with and / or binds to the same epitope as an antibody comprising the heavy chain variable domain of SEQ ID NO: 21 and the light chain variable domain of SEQ ID NO: 22.

[0201] In various embodiments, the antibody or antigen-binding fragment (Ab) in the ADC is an anti-EPHA2 antibody or its antigen-binding fragment. In some embodiments, the antibody or antigen-binding fragment binds to EPHA2 and targets EPHA2-expressing neoplasms (i.e., the ADC targets EPHA2-expressing neoplasms). In some embodiments, the antibody or antigen-binding fragment in the ADC is an internalized anti-EPHA2 antibody or its internalized antigen-binding fragment.

[0202] In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing 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 an internalizing antigen-binding fragment. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises a human frame sequence. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:23 and a light chain variable region containing 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 Igκ or λ light chain constant region. In some embodiments, the anti-EPHA2 antibody or antigen-binding fragment competes with 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.

[0203] In various embodiments, this document also provides compounds comprising a linker-drug as defined by the general formula LD, where L = linker portion and D = drug portion (e.g., splice modulator drug portion). In various embodiments, the linker-drug (LD) compounds disclosed herein may be linked to antibody or antigen-binding fragments and / or adapted to the ADCs disclosed herein, such as ADCs having formula (I).

[0204] In various embodiments, the connector-drug (LD) compounds disclosed herein comprise a connector-drug structure according to formula (III). In various embodiments, this disclosure provides connector-drug (LD) compounds having formula (III):

[0205] or a pharmaceutically acceptable salt thereof, wherein:

[0206] R 1Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0207] R 2 It may not exist or may be a connector;

[0208] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[0209] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0210] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[0211] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and

[0212] -C(=O)-OR 17 ;

[0213] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0214] Z is selected from

[0215] where R 1 R 2 R 3R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[0216] where R 6 and R 7 At least one of them is hydrogen; and

[0217] Where R 2 If it is a connector, then R 6 and R 7 None of them are connectors, and if R 6 or R 7 If it is a connector, then R 2 Does not exist.

[0218] In various other embodiments, the connector-drug (LD) compounds disclosed herein comprise a connector-drug structure according to formula (V). In different embodiments, this disclosure provides connector-drug (LD) compounds having formula (V):

[0219] or a pharmaceutically acceptable salt thereof, wherein:

[0220] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0221] R 2 It may not exist or may be a connector;

[0222] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[0223] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0224] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[0225] R 15 and R 16 Each is independently selected from hydrogen, R 17 -C(=O)-R 17 and -C(=O)-OR 17 ;and

[0226] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0227] Where R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[0228] Where R 6 and R 7 At least one of them is hydrogen; and

[0229] Where R 2 If it is a connector, then R 6 and R 7 None of them are connectors, and if R6 or R 7 If it is a connector, then R 2 Does not exist.

[0230] In various other embodiments, the connector-drug (LD) compounds disclosed herein comprise a connector-drug structure according to formula (VII). In different embodiments, this disclosure provides connector-drug (LD) compounds having formula (VII):

[0231] or a pharmaceutically acceptable salt thereof, wherein:

[0232] R 1 and R 9 Each is independently selected from the absence of, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0233] R 2 It may not exist or may be a connector;

[0234] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[0235] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0236] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[0237] R 10 Selected from hydrogen, C1-C6 alkyl groups, -C(=O)-(C1-C6 alkyl) groups, and -CD3;

[0238] R 15 and R 16 Each is independently selected from hydrogen, R 17 -C(=O)-R 17 and

[0239] -C(=O)-OR 17 ;

[0240] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0241] a is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0242] Where 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 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[0243] Where R 6 and R 7 At least one of them is hydrogen;

[0244] Where R 2 If it is a connector, then R 6 and R 7 None of them are connectors, and if R 6 Or R 7 If it is a connector, then R 2 It does not exist; and

[0245] Where R 1 and R 9 They cannot both be absent at the same time.

[0246] In various other embodiments, the connector-drug (LD) compounds disclosed herein comprise a connector-drug structure according to formula (IX). In different embodiments, this disclosure provides connector-drug (LD) compounds having formula (IX):

[0247] or a pharmaceutically acceptable salt thereof, wherein:

[0248] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0249] R 2 It's a connector;

[0250] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[0251] R 4 Selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0252] R 10 Selected from 3- to 10-membered carbon rings and 3- to 10-membered heterocycles, each of which is composed of 0 to 3 R... a Replace, where each R a Independently selected from halogens, C1-C6 alkyl groups, -O-(C1-C6)alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylhydroxy groups, and -S (=O). w -(4- to 7-membered heterocycles), 4- to 7-membered carbon rings and 4- to 7-membered heterocycles;

[0253] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and -C(=O)-OR 17 ;and

[0254] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0255] Where R 1 R 2 R 3 R 4 R 10 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0256] Each R a Independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, -NR 15 R 16 C1-C6 alkyl groups, -(C=O)-(C1-C6 alkyl) groups, -(C=O)-(C1-C6 alkyl)-(C3-C 10 The heterocyclic group and the C1-C6 alkylcarboxylic acid group are each substituted by 0, 1, or 2 independently selected groups from the following: halogen, hydroxyl group, -NR 15 R 16 and C1-C3 alkyl groups; and

[0257] w is 0, 1, or 2.

[0258] Additionally, in various embodiments, this document provides for the therapeutic use of the said ADC complex and composition, for example, in treating aplastic conditions (e.g., cancer). In some aspects, this disclosure provides a method for treating aplastic conditions, such as cancers expressing an antigen targeted by an antibody or antigen-binding fragment in the ADC, such as HER2, CD138, EPHA2, MSLN, FOLH1, CDH6, CEACAM5, CFC1B, ENPP3, FOLR1, HAVCR1, KIT, MET, MUC16, SLC39A6, SLC44A4, or STEAP1.

[0259] In some aspects, this disclosure provides a method of treating a subject with or suspected of having a neoplastic condition by administering to a subject a therapeutically effective amount and / or regimen of any of the ADC or composition. In some embodiments, the neoplastic condition is a hematologic malignancy or a solid tumor. In some embodiments, the neoplastic condition is a hematologic malignancy. In some embodiments, the hematologic malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma. In some embodiments, the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the neoplastic condition 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 gland duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney 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.

[0260] In some embodiments, treatment with the antibody-drug conjugate or composition induces bystander killing of vesicular cells that do not express the target antigen but are adjacent to vesicular cells that express the target antigen. In some embodiments, the subject has one or more vesicular cells that express the target antigen.

[0261] In some embodiments, the target antigen is HER2. In some embodiments, the one or more proliferative cells are derived from breast cancer, ovarian cancer, gastric cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial cancer), osteosarcoma, or salivary gland duct carcinoma expressing HER2. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-HER2 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0262] In some embodiments, the target antigen is CD138. In some embodiments, the one or more proliferative cells are derived from multiple myeloma expressing CD138. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-CD138 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0263] In some embodiments, the target antigen is EPHA2. In some embodiments, the one or more proliferative cells are derived from breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-EPHA2 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0264] In some embodiments, the target antigen is MSLN. In some embodiments, the one or more proliferative cells are derived from ovarian cancer, cervical cancer, pancreatic cancer, or lung cancer (e.g., lung adenocarcinoma) expressing MSLN. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-MSLN antibody when administered alone and / or (b) a splice modulator when administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator when administered alone.

[0265] In some embodiments, the target antigen is FOLH1. In some embodiments, the one or more proliferative cells are derived from prostate cancer expressing FOLH1. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-FOLH1 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0266] In some embodiments, the target antigen is CDH6. In some embodiments, the one or more proliferative cells are derived from renal cell carcinoma expressing CDH6. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-CDH6 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0267] In some embodiments, the target antigen is CEACAM5. In some embodiments, the one or more proliferative cells are derived from colorectal cancer expressing CEACAM5. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-CEACAM5 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0268] In some embodiments, the target antigen is CFC1B. In some embodiments, the one or more proliferative cells are derived from pancreatic cancer expressing CFC1B. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-CFC1B antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0269] In some embodiments, the target antigen is ENPP3. In some embodiments, the one or more proliferative cells are derived from renal cell carcinoma expressing ENPP3. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-ENPP3 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0270] In some embodiments, the target antigen is FOLR1. In some embodiments, the one or more proliferative cells are derived from ovarian cancer expressing FOLR1. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-FOLR1 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0271] In some embodiments, the target antigen is HAVCR1. In some embodiments, the one or more proliferative cells are derived from renal or esophageal cancer expressing HAVCR1. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-HAVCR1 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0272] In some embodiments, the target antigen is KIT. In some embodiments, the one or more proliferative cells are derived from renal cell carcinoma expressing KIT. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-KIT antibody administered alone and / or (b) a splice modulator administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator administered alone.

[0273] In some embodiments, the target antigen is MET. In some embodiments, the one or more proliferative cells are derived from renal or esophageal cancer expressing MET. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-MET antibody administered alone and / or (b) a splice modulator administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator administered alone.

[0274] In some embodiments, the target antigen is MUC16. In some embodiments, the one or more proliferative cells are derived from ovarian cancer, cervical cancer, or breast cancer expressing MUC16. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-MUC16 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0275] In some embodiments, the target antigen is SLC39A6. In some embodiments, the one or more proliferative cells are derived from breast or prostate cancer expressing SLC39A6. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-SLC39A6 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0276] In some embodiments, the target antigen is SLC44A4. In some embodiments, the one or more proliferative cells are derived from prostate cancer expressing SLC44A4. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-SLC44A4 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0277] In some embodiments, the target antigen is STEAP1. In some embodiments, the one or more proliferative cells are derived from prostate cancer expressing STEAP1. In some embodiments, the subject is unresponsive or poorly responsive to treatment with (a) an anti-STEAP1 antibody as administered alone and / or (b) a splice modulator as administered alone. In some embodiments, the subject is intolerant, unresponsive, or poorly responsive to treatment with a splice modulator as administered alone.

[0278] In some other respects, this disclosure provides a method for reducing or inhibiting tumor growth in subjects who have or are suspected of having aplastic disease by administering to a subject a therapeutically effective amount and / or regimen of any of the ADCs or compositions.

[0279] 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 are adjacent to neoplastic tumor cells that express the target antigen. In some embodiments, the tumor comprises one or more neoplastic cells expressing the target antigen.

[0280] In some embodiments, the target antigen is HER2. In some embodiments, the one or more proliferative cells are derived from breast cancer, ovarian cancer, gastric cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial cancer), osteosarcoma, or salivary gland duct carcinoma that express HER2. 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.

[0281] In some embodiments, the target antigen is CD138. In some embodiments, the one or more proliferative cells are derived from multiple myeloma expressing 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.

[0282] In some embodiments, the target antigen is EPHA2. In some embodiments, the one or more proliferative 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 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.

[0283] In some embodiments, the target antigen is MSLN. In some embodiments, the one or more proliferative cells are derived from ovarian cancer, cervical cancer, pancreatic cancer, or lung cancer (e.g., lung adenocarcinoma) expressing MSLN. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-MSLN antibody when administered alone and / or (b) a splice modulator when administered alone.

[0284] In some embodiments, the target antigen is FOLH1. In some embodiments, the one or more proliferative cells are derived from prostate cancer expressing 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.

[0285] In some embodiments, the target antigen is CDH6. In some embodiments, the one or more proliferative cells are derived from renal cell carcinoma expressing CDH6. 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.

[0286] In some embodiments, the target antigen is CEACAM5. In some embodiments, the one or more proliferative cells are derived from colorectal cancer expressing 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 splice modulator when administered alone.

[0287] In some embodiments, the target antigen is CFC1B. In some embodiments, the one or more proliferative cells are derived from pancreatic cancer expressing 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.

[0288] In some embodiments, the target antigen is ENPP3. In some embodiments, the one or more proliferative cells are derived from renal cell carcinoma expressing 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.

[0289] In some embodiments, the target antigen is FOLR1. In some embodiments, the one or more proliferative cells are derived from ovarian cancer expressing 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.

[0290] In some embodiments, the target antigen is HAVCR1. In some embodiments, the one or more proliferative cells are derived from renal or esophageal cancer expressing 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.

[0291] In some embodiments, the target antigen is KIT. In some embodiments, the one or more proliferative cells are derived from renal cell carcinoma expressing KIT. In some embodiments, the tumor is resistant or refractory to treatment with (a) an anti-KIT antibody administered alone and / or (b) a splice modulator administered alone.

[0292] In some embodiments, the target antigen is MET. In some embodiments, the one or more proliferative cells are derived from renal or esophageal cancer expressing MET. 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.

[0293] In some embodiments, the target antigen is MUC16. In some embodiments, the one or more proliferative cells are derived from ovarian cancer, cervical cancer, or breast cancer expressing 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.

[0294] In some embodiments, the target antigen is SLC39A6. In some embodiments, the one or more proliferative cells are derived from breast or prostate cancer expressing 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.

[0295] In some embodiments, the target antigen is SLC44A4. In some embodiments, the one or more proliferative cells are derived from prostate cancer expressing 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.

[0296] In some embodiments, the target antigen is STEAP1. In some embodiments, the one or more proliferative cells are derived from prostate cancer expressing 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.

[0297] In other aspects, this disclosure provides a method for determining whether a subject with or suspected of having a neoplastic condition will respond to treatment with any of the ADC or composition by providing a biological sample from a 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, blood fractions, or cells obtained from the blood or blood fraction. In some embodiments, the subject has one or more neoplastic cells expressing a target antigen. In some embodiments, the target antigen is HER2. In some embodiments, the one or more neoplastic cells are derived from breast cancer, ovarian cancer, gastric cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial cancer), osteosarcoma, or salivary gland duct carcinoma expressing HER2. In some embodiments, the target antigen is CD138. In some embodiments, the one or more neoplastic cells are derived from multiple myeloma expressing CD138. In some embodiments, the target antigen is EPHA2. In some embodiments, the one or more proliferative cells are derived from breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2. In some embodiments, the target antigen is MSLN. In some embodiments, the one or more proliferative cells are derived from ovarian cancer, cervical cancer, pancreatic cancer, or lung cancer (e.g., lung adenocarcinoma) expressing MSLN. In some embodiments, the target antigen is FOLH1. In some embodiments, the one or more proliferative cells are derived from prostate cancer expressing FOLH1. In some embodiments, the target antigen is CDH6. In some embodiments, the one or more proliferative cells are derived from renal cancer expressing CDH6. In some embodiments, the target antigen is CEACAM5. In some embodiments, the one or more proliferative cells are derived from colorectal cancer expressing CEACAM5. In some embodiments, the target antigen is CFC1B. In some embodiments, the one or more proliferative cells are derived from pancreatic cancer expressing CFC1B. In some embodiments, the target antigen is ENPP3. In some embodiments, the one or more proliferative cells are derived from renal cancer expressing ENPP3. In some embodiments, the target antigen is FOLR1. In some embodiments, the one or more proliferative cells are derived from ovarian cancer expressing FOLR1. In some embodiments, the target antigen is HAVCR1. In some embodiments, the one or more proliferative cells are derived from renal or esophageal cancer expressing HAVCR1. In some embodiments, the target antigen is KIT. In some embodiments, the one or more proliferative cells are derived from renal cancer expressing KIT. In some embodiments, the target antigen is MET. In some embodiments, the one or more proliferative cells are derived from renal or esophageal cancer expressing MET. 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 expressing MUC16. In some embodiments, the target antigen is SLC39A6. In some embodiments, the one or more neoplastic cells are derived from breast or prostate cancer expressing SLC39A6. In some embodiments, the target antigen is SLC44A4. In some embodiments, the one or more neoplastic cells are derived from prostate cancer expressing SLC44A4. In some embodiments, the target antigen is STEAP1. In some embodiments, the one or more neoplastic cells are derived from prostate cancer expressing STEAP1.

[0298] In various embodiments, this document further provides pharmaceutical compositions comprising an ADC and a pharmaceutically acceptable diluent, carrier, and / or excipient. Methods for manufacturing the said ADC complex and compositions are also disclosed.

[0299] This application relates to the following:

[0300] Item 1. An antibody-drug conjugate having formula (I),

[0301] Ab-(LD) p (I)

[0302] in

[0303] Ab is an antibody or antigen-binding fragment that targets neoplastic cells;

[0304] D is a splice modifier;

[0305] L is the connector that covalently links Ab to D; and

[0306] p is an integer from 1 to 15.

[0307] Item 2. The antibody-drug conjugate as described in Item 1, wherein the antibody or antigen-binding fragment targets cytoplasmic cells derived from hematologic malignancies or solid tumors.

[0308] Item 3. The antibody-drug conjugate as described in Item 1 or Item 2, wherein the antibody or antigen-binding fragment targets cytoplasmic cells derived from hematologic malignancies.

[0309] Item 4. The antibody-drug conjugate as described in Item 3, wherein the hematologic malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

[0310] Item 5. The antibody-drug conjugate as described in Item 3 or Item 4, wherein the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma.

[0311] Item 6. The antibody-drug conjugate as described in Item 1 or Item 2, wherein the antibody or antigen-binding fragment targets neoplastic cells derived from solid tumors.

[0312] Item 7. The antibody-drug conjugate as described in Item 6, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary gland duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney cancer, colorectal cancer, and esophageal cancer.

[0313] Item 8. An antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing HER2.

[0314] Item 9. The antibody-drug conjugate as described in Item 8, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

[0315] Item 10. The antibody-drug conjugate as described in Item 8 or Item 9, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[0316] Item 11. The antibody-drug conjugate as described in any one of items 8 to 10, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[0317] Item 12. The antibody-drug conjugate as described in any one of items 8 to 11, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[0318] Item 13. The antibody-drug conjugate as described in any one of items 8 to 12, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0319] Item 14. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing CD138.

[0320] Item 15. The antibody-drug conjugate as described in Item 14, wherein the antibody or antigen-binding fragment is an anti-CD138 antibody or antigen-binding fragment.

[0321] Item 16. The antibody-drug conjugate as described in Item 14 or Item 15, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

[0322] Item 17. The antibody-drug conjugate as described in any one of items 14 to 16, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:21 and a light chain variable region containing the amino acid sequence of SEQ ID NO:22.

[0323] Item 18. The antibody-drug conjugate as described in any one of items 14 to 17, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG2a heavy chain.

[0324] Item 19. The antibody-drug conjugate as described in any one of items 14 to 18, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0325] Item 20. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing EPHA2.

[0326] Item 21. The antibody-drug conjugate as described in Item 20, wherein the antibody or antigen-binding fragment is an anti-EPHA2 antibody or antigen-binding fragment.

[0327] Item 22. The antibody-drug conjugate as described in Item 20 or Item 21, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:16 (LCDR1), SEQ ID NO:17 (LCDR2), and SEQ ID NO:18 (LCDR3).

[0328] Item 23. The antibody-drug conjugate as described in any one of items 20 to 22, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:23 and a light chain variable region containing the amino acid sequence of SEQ ID NO:24.

[0329] Item 24. The antibody-drug conjugate as described in any one of items 20 to 23, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[0330] Item 25. The antibody-drug conjugate as described in any one of items 20 to 24, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0331] Item 26. The antibody-drug conjugate as described in any one of items 1 to 25, wherein the linker is a cleavable linker.

[0332] Item 27. The antibody-drug conjugate as described in Item 26, wherein the linker comprises a cleavable peptide moiety.

[0333] Item 28. The antibody-drug conjugate as described in Item 27, wherein the cleavable peptide moiety is cleavable by an enzyme.

[0334] Item 29. The antibody-drug conjugate as described in any one of items 26 to 28, wherein the cleavable peptide portion or linker comprises an amino acid unit.

[0335] Item 30. The antibody-drug conjugate as described in Item 29, wherein the amino acid unit comprises valine-citrulline (Val-Cit).

[0336] Item 31. The antibody-drug conjugate as described in Item 29, wherein the amino acid unit comprises valine-alanine (Val-Ala).

[0337] Item 32. The antibody-drug conjugate as described in Item 26, wherein the linker comprises a cleavable glucuronide moiety.

[0338] Item 33. The antibody-drug conjugate as described in Item 32, wherein the cleavable glucuronide moiety is cleavable by an enzyme.

[0339] Item 34. The antibody-drug conjugate as described in Item 32 or Item 33, wherein the cleavable glucuronide moiety is cleaved by glucuronidase.

[0340] Item 35. The antibody-drug conjugate as described in any one of items 32 to 34, wherein the cleavable glucuronide moiety is cleaved by β-glucuronidase.

[0341] Item 36. The antibody-drug conjugate as described in any one of items 1 to 35, wherein the linker comprises at least one spacer subunit.

[0342] Item 37. The antibody-drug conjugate as described in Item 36, wherein the spacer subunit or linker comprises a polyethylene glycol (PEG) portion.

[0343] Item 38. The antibody-drug conjugate as described in Item 37, wherein the PEG portion comprises -(PEG). m - And m is an integer from 1 to 10.

[0344] Item 39. The antibody-drug conjugate as described in Item 38, wherein m is 2.

[0345] Item 40. The antibody-drug conjugate as described in Item 36, wherein the spacer subunit or linker comprises an alkyl portion.

[0346] Item 41. The antibody-drug conjugate as described in Item 40, wherein the alkyl moiety comprises -(CH2). n - and n is an integer from 1 to 10.

[0347] Item 42. The antibody-drug conjugate as described in Item 41, wherein n is 2.

[0348] Item 43. The antibody-drug conjugate as described in Item 41, wherein n is 5.

[0349] Item 44. The antibody-drug conjugate as described in Item 41, wherein n is 6.

[0350] Item 45. An antibody-drug conjugate as described in any one of items 36 to 44, wherein the spacer unit is linked to the antibody or antigen-binding fragment via a maleimide (Mal) portion (“Mal-spacer unit”).

[0351] Item 46. The antibody-drug conjugate as described in Item 45, wherein the Mal-spacer unit is reactive with cysteine ​​residues on the antibody or antigen-binding fragment.

[0352] Item 47. The antibody-drug conjugate as described in Item 45 or Item 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.

[0353] Item 48. The antibody-drug conjugate as described in any one of items 45 to 47, wherein the linker comprises the Mal-spacer subunit and the cleavable peptide moiety.

[0354] Item 49. The antibody-drug conjugate as described in Item 48, wherein the cleavable peptide portion comprises amino acid units.

[0355] Item 50. The antibody-drug conjugate as described in Item 48 or 49, wherein the cleavable peptide portion or amino acid unit comprises Val-Cit.

[0356] Item 51. The antibody-drug conjugate as described in Item 48 or Item 49, wherein the cleavable peptide moiety or amino acid unit comprises Val-Ala.

[0357] Item 52. The antibody-drug conjugate as described in any one of items 45 to 51, wherein the Mal-spacer unit comprises an alkyl moiety.

[0358] Item 53. The antibody-drug conjugate as described in any one of items 45 to 51, wherein the Mal-spacer subunit comprises a PEG portion.

[0359] Item 54. The antibody-drug conjugate as described in any one of items 45 to 52, wherein the Mal-spacer subunit comprises maleimide hexanoyl (MC).

[0360] Item 55. The antibody-drug conjugate as described in any one of items 45 to 54, wherein the Mal-spacer subunit links the antibody or antigen-binding fragment to the cleavable portion in the adapter.

[0361] Item 56. The antibody-drug conjugate as described in Item 55, wherein the cleavable portion of the linker comprises a cleavable peptide portion.

[0362] Item 57. The antibody-drug conjugate as described in Item 56, wherein the cleavable peptide portion comprises amino acid units.

[0363] Item 58. The antibody-drug conjugate as described in Item 56 or Item 57, wherein the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn.

[0364] Item 59. The antibody-drug conjugate as described in any one of items 55 to 58, wherein the linker comprises MC-Val-Cit.

[0365] Item 60. The antibody-drug conjugate as described in any one of items 55 to 58, wherein the linker comprises MC-Val-Ala.

[0366] Item 61. The antibody-drug conjugate as described in any one of items 55 to 60, wherein the Mal-spacer unit comprises an alkyl moiety.

[0367] Item 62. The antibody-drug conjugate as described in any one of items 55 to 60, wherein the Mal-spacer subunit comprises a PEG portion.

[0368] Item 63. The antibody-drug conjugate as described in any one of items 55 to 61, wherein the Mal-spacer subunit comprises maleimide hexanoyl (MC).

[0369] Item 64. The antibody-drug conjugate of any one of items 36 to 63, wherein the cleavable portion of the connector is directly bound to the splice modulator, or wherein a spacer subunit connects the cleavable portion of the connector to the splice modulator.

[0370] Item 65. The antibody-drug conjugate as described in Item 64, wherein cleavage of the conjugate releases the splicing regulator from the antibody or antigen-binding fragment and the linker.

[0371] Item 66. The antibody-drug conjugate as described in Item 64 or Item 65, wherein the spacer subunit to which the cleavable portion of the linker is attached to the splice modulator is self-ablating.

[0372] Item 67. The antibody-drug conjugate as described in any one of items 64 to 66, wherein the spacer subunit of the linker to which the cleavable portion of the linker is attached comprises a p-aminobenzyloxycarbonyl group (pABC).

[0373] Item 68. The antibody-drug conjugate as described in Item 67, wherein the pABC connects the cleavable portion of the linker to the splice modulator.

[0374] Item 69. The antibody-drug conjugate as described in Item 67 or Item 68, wherein the cleavable portion of the linker comprises a cleavable peptide portion.

[0375] Item 70. The antibody-drug conjugate as described in Item 69, wherein the cleavable peptide portion comprises amino acid units.

[0376] Item 71. The antibody-drug conjugate as described in Item 69 or Item 70, wherein the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn.

[0377] Item 72. The antibody-drug conjugate as described in any one of items 67 to 71, wherein the linker comprises Val-Cit-pABC.

[0378] Item 73. The antibody-drug conjugate as described in any one of items 67 to 71, wherein the linker comprises Val-Ala-pABC.

[0379] Item 74. The antibody-drug conjugate as described in any one of items 64 to 66, wherein the spacer subunit of the linker to which the cleavable portion of the linker is attached comprises p-aminobenzyl (pAB).

[0380] Item 75. The antibody-drug conjugate as described in Item 74, wherein the pAB connects the cleavable portion of the linker to the splice modulator.

[0381] Item 76. The antibody-drug conjugate as described in Item 74 or Item 75, wherein the cleavable portion of the linker comprises a cleavable peptide portion.

[0382] Item 77. The antibody-drug conjugate as described in Item 76, wherein the cleavable peptide portion comprises amino acid units.

[0383] Item 78. The antibody-drug conjugate as described in Item 76 or Item 77, wherein the cleavable peptide moiety or amino acid unit comprises Val-Cit, Val-Ala, Glu-Val-Cit, or Ala-Ala-Asn.

[0384] Item 79. The antibody-drug conjugate as described in any one of items 74 to 78, wherein the linker comprises Val-Cit-pAB.

[0385] Item 80. The antibody-drug conjugate as described in any one of items 74 to 78, wherein the linker comprises Val-Ala-pAB.

[0386] Item 81. The antibody-drug conjugate as described in any one of items 1 to 25, wherein the linker is a non-cleavable linker.

[0387] Item 82. The antibody-drug conjugate as described in Item 81, wherein the linker comprises at least one spacer subunit.

[0388] Item 83. The antibody-drug conjugate as described in Item 81 or Item 82, wherein the spacer subunit or linker comprises a polyethylene glycol (PEG) portion.

[0389] Item 84. The antibody-drug conjugate as described in Item 83, wherein the PEG portion comprises -(PEG). m - And m is an integer from 1 to 10.

[0390] Item 85. The antibody-drug conjugate as described in Item 84, wherein m is 2.

[0391] Item 86. The antibody-drug conjugate as described in Item 81 or Item 82, wherein the spacer subunit or linker comprises an alkyl portion.

[0392] Item 87. The antibody-drug conjugate as described in Item 86, wherein the alkyl moiety comprises -(CH2). n - and n is an integer from 1 to 10.

[0393] Item 88. The antibody-drug conjugate as described in Item 87, wherein n is 2.

[0394] Item 89. The antibody-drug conjugate as described in Item 87, wherein n is 5.

[0395] Item 90. The antibody-drug conjugate as described in Item 87, wherein n is 6.

[0396] Item 91. An antibody-drug conjugate as described in any one of items 81 to 90, wherein the spacer subunit is linked to the antibody or antigen-binding fragment via a maleimide (Mal) portion (“Mal-spacer subunit”).

[0397] Item 92. The antibody-drug conjugate as described in Item 91, wherein the Mal-spacer unit is reactable with cysteine ​​residues on the antibody or antigen-binding fragment.

[0398] Item 93. The antibody-drug conjugate as described in Item 91 or Item 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.

[0399] Item 94. The antibody-drug conjugate as described in any one of items 91 to 93, wherein the Mal-spacer unit comprises an alkyl moiety.

[0400] Item 95. The antibody-drug conjugate as described in any one of items 91 to 93, wherein the Mal-spacer subunit comprises a PEG portion.

[0401] Item 96. The antibody-drug conjugate as described in any one of items 91 to 95, wherein the linker or Mal-spacer subunit comprises maleimide hexanoyl (MC).

[0402] Item 97. The antibody-drug conjugate as described in Item 96, wherein the linker or Mal-spacer unit comprises maleimide hexanoyl (MC) and at least one additional spacer unit.

[0403] Item 98. The antibody-drug conjugate as described in any one of items 91 to 95, wherein the linker or Mal-spacer subunit comprises MC-(PEG)2.

[0404] Item 99. The antibody-drug conjugate as described in Item 98, wherein the linker or Mal-spacer subunit comprises MC-(PEG)2 and at least one additional spacer subunit.

[0405] Item 100. The antibody-drug conjugate as described in any one of items 91 to 95, wherein the linker or Mal-spacer subunit comprises Mal-Hex.

[0406] Item 101. The antibody-drug conjugate as described in Item 100, wherein the linker or Mal-spacer subunit comprises MC-Hex and at least one additional spacer subunit.

[0407] Item 102. The antibody-drug conjugate as described in any one of items 91 to 95, wherein the linker or Mal-spacer subunit comprises Mal-Et.

[0408] Item 103. The antibody-drug conjugate as described in Item 102, wherein the linker or Mal-spacer subunit comprises Mal-Et and at least one additional spacer subunit.

[0409] Item 104. The antibody-drug conjugate as described in any one of items 91 to 95, wherein the linker or Mal-spacer subunit comprises Mal-Et-O-Et.

[0410] Item 105. The antibody-drug conjugate as described in Item 104, wherein the linker or Mal-spacer subunit comprises Mal-Et-O-Et and at least one additional spacer subunit.

[0411] Item 106. The antibody-drug conjugate as described in any one of items 91 to 105, wherein the Mal-spacer subunit links the antibody or antigen-binding fragment to the splicing regulator.

[0412] Item 107. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator comprises a splicing modulator having formula (II):

[0413] or a pharmaceutically acceptable salt thereof, wherein:

[0414] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0415] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[0416] R 4 R 5 and R 8Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0417] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups and -NR 15 R 16 ;

[0418] R 15 and R 16 Each is independently selected from hydrogen, R 17 -C(=O)-R 17 and -C(=O)-OR 17 ;

[0419] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0420] Z is selected from

[0421] Where R 1 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups.

[0422] Where R 6 and R 7 At least one of them is hydrogen.

[0423] Item 108. The antibody-drug conjugate as described in Item 107, wherein the linker is covalently linked to the splice modulator (“LD”), and the LD has a structure of formula (II-A):

[0424] Or its pharmaceutically acceptable salt.

[0425] Z' is selected from and

[0426] All other variables are defined as per equation (II).

[0427] Item 109. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator is a splicing modulator having formula (IV):

[0428] or a pharmaceutically acceptable salt thereof, wherein:

[0429] R 1 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, -OC(=O)-(C1-C6 alkyl) groups, and -CD3;

[0430] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[0431] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0432] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups and -NR 15 R 16 ;

[0433] R 15 and R 16 Each is independently selected from hydrogen, R 17 -C(=O)-R 17and -C(=O)-OR 17 ;and

[0434] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0435] where R 1 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups.

[0436] where R 6 and R 7 At least one of them is hydrogen.

[0437] Item 110. The antibody-drug conjugate as described in Item 109, wherein the linker is covalently linked to the splice modulator (“LD”), and the LD has a structure of formula (IV-A):

[0438] Or its pharmaceutically acceptable salt.

[0439] Item 111. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator is a splicing modulator having formula (VI):

[0440] or a pharmaceutically acceptable salt thereof, wherein:

[0441] R 1 and R 9 Each group is independently selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, -OC(=O)-(C1-C6 alkyl) groups, and -CD3;

[0442] R 3Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[0443] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0444] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[0445] R 10 Selected from hydrogen, C1-C6 alkyl groups, -C(=O)-(C1-C6 alkyl) groups, and -CD3;

[0446] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and -C(=O)-OR 17 ;

[0447] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0448] a is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0449] where R 1 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 15 R 16 and R 17Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[0450] where R 6 and R 7 At least one of them is hydrogen; and

[0451] where R 1 and R 9 They cannot both be absent at the same time.

[0452] Item 112. The antibody-drug conjugate as described in Item 111, wherein the linker is covalently linked to the splice modulator (“LD”), and the LD has a structure of formula (VI-A):

[0453] Or its pharmaceutically acceptable salt.

[0454] Item 113. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator is a splicing modulator having formula (VIII):

[0455] or a pharmaceutically acceptable salt thereof, wherein:

[0456] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0457] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[0458] R 4 Selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups; and

[0459] R 10Selected from 3- to 10-membered carbon rings and 3- to 10-membered heterocycles, each of which is composed of 0 to 3 R... a Replace, where each R a Independently selected from halogens, C1-C6 alkyl groups, -O-(C1-C6)alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylhydroxy groups, and -S (=O). w -(4- to 7-membered heterocycles), 4- to 7-membered carbon rings and 4- to 7-membered heterocycles;

[0460] R 15 and R 16 Each is independently selected from hydrogen, R 17 -C(=O)-R 17 and -C(=O)-OR 17 ;and

[0461] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0462] Where R 1 R 3 R 4 R 10 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0463] Each R a Independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, -NR 15 R 16 C1-C6 alkyl groups, -(C=O)-(C1-C6 alkyl) groups, -(C=O)-(C1-C6 alkyl)-(C3-C 10 Heterocyclic groups, -S (=O) w -(C3-C8 heterocyclic group) and C1-C6 alkylcarboxylic acid group, each of which is substituted by 0, 1 or 2 independently selected from the following groups: halogen, hydroxyl group, -NR 15 R 16 and C1-C3 alkyl groups; and

[0464] w is 0, 1, or 2.

[0465] Item 114. The antibody-drug conjugate as described in Item 113, wherein the linker is covalently linked to the splice modulator (“LD”), and the LD has a structure of formula (VIII-A):

[0466] Or its pharmaceutically acceptable salt.

[0467] Item 115. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator comprises a modulator of the SF3b complex.

[0468] Item 116. The antibody-drug conjugate as described in Item 115, wherein the splicing modulator comprises prasadiene lactone or a prasadiene lactone derivative.

[0469] Item 117. The antibody-drug conjugate as described in Item 115 or Item 116, wherein the splicing modulator comprises prasadiene lactone D or a prasadiene lactone D derivative.

[0470] Item 118. The antibody-drug conjugate as described in Item 117, wherein the prasadiene lactone D or its derivative comprises D2, D1, D4, D8, D10, D11 (E7107), D20, D21, D22, D12 or D25.

[0471] Item 119. The antibody-drug conjugate as described in Item 117 or Item 118, wherein the prasadiene lactone D or its derivative comprises D2.

[0472] Item 120. The antibody-drug conjugate as described in Item 117 or Item 118, wherein the prasadiene lactone D or its derivative comprises D1.

[0473] Item 121. The antibody-drug conjugate as described in Item 117 or Item 118, wherein the prasadiene lactone D or its derivative is a zwitterionic prasadiene lactone D or its derivative.

[0474] Item 122. The antibody-drug conjugate as described in Item 121, wherein the zwitterionic prasadiene lactone D or its derivative comprises D22 or D25.

[0475] Item 123. The antibody-drug conjugate as described in Item 115 or Item 116, wherein the splicing modulator comprises prasadiene lactone B or a prasadiene lactone B derivative.

[0476] Item 124. The antibody-drug conjugate as described in Item 123, wherein the prasadiene lactone B or its derivative comprises D9, D18, D19 or D13.

[0477] Item 125. The antibody-drug conjugate as described in Item 115 or Item 116, wherein the splicing modulator comprises arylpradolide lactone.

[0478] Item 126. The antibody-drug conjugate as described in Item 125, wherein the arylpradoside comprises D15, D14, D16, D17, D26, or D33.

[0479] Item 127. The antibody-drug conjugate as described in Item 125 or Item 126, wherein the arylpradolide is a zwitterionic arylpradolide.

[0480] Item 128. The antibody-drug conjugate as described in Item 127, wherein the zwitterionic arylpradolide comprises D33.

[0481] Item 129. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator comprises D2:

[0482]

[0483] Item 130. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator comprises D1:

[0484]

[0485] Item 131. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator comprises D4, D12, D15, D8, D9, D10, D13, D18, D19, D20, D21, D22, D25 or D33.

[0486] Item 132. The antibody-drug conjugate as described in any one of items 1 to 131, wherein p is 1 to 10.

[0487] Item 133. The antibody-drug conjugate as described in any one of items 1 to 132, wherein p is 2 to 8.

[0488] Item 134. The antibody-drug conjugate as described in any one of items 1 to 133, wherein p is 4 to 8.

[0489] Item 135. An antibody-drug conjugate as described in any one of items 1 to 134, wherein p is 4.

[0490] Item 136. An antibody-drug conjugate as described in any one of items 1 to 134, wherein p is 8.

[0491] Item 137. An antibody-drug conjugate having formula (I),

[0492] Ab-(LD) p (I)

[0493] in

[0494] Ab is an antibody or antigen-binding fragment that targets neoplastic cells;

[0495] D is D2;

[0496] L is the connector that covalently links Ab to D; and

[0497] p is an integer from 1 to 15.

[0498] Item 138. The antibody-drug conjugate as described in Item 137, wherein the antibody or antigen-binding fragment targets cells expressing HER2.

[0499] Item 139. The antibody-drug conjugate as described in Item 138, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

[0500] Item 140. The antibody-drug conjugate as described in Item 138 or Item 139, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[0501] Item 141. The antibody-drug conjugate as described in any one of items 138 to 140, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[0502] Item 142. The antibody-drug conjugate as described in any one of items 138 to 141, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[0503] Item 143. The antibody-drug conjugate as described in any one of items 138 to 142, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0504] Item 144. The antibody-drug conjugate as described in Item 137, wherein the antibody or antigen-binding fragment targets cells expressing CD138.

[0505] Item 145. The antibody-drug conjugate as described in Item 144, wherein the antibody or antigen-binding fragment is an anti-CD138 antibody or antigen-binding fragment.

[0506] Item 146. The antibody-drug conjugate as described in Item 144 or Item 145, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

[0507] Item 147. The antibody-drug conjugate as described in any one of items 144 to 146, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:21 and a light chain variable region containing the amino acid sequence of SEQ ID NO:22.

[0508] Item 148. The antibody-drug conjugate as described in any one of items 144 to 147, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG2a heavy chain.

[0509] Item 149. The antibody-drug conjugate as described in any one of items 144 to 148, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0510] Item 150. The antibody-drug conjugate as described in Item 137, wherein the antibody or antigen-binding fragment targets cells expressing EPHA2.

[0511] Item 151. The antibody-drug conjugate as described in Item 150, wherein the antibody or antigen-binding fragment is an anti-EPHA2 antibody or antigen-binding fragment.

[0512] Item 152. The antibody-drug conjugate as described in Item 150 or Item 151, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:16 (LCDR1), SEQ ID NO:17 (LCDR2), and SEQ ID NO:18 (LCDR3).

[0513] Item 153. The antibody-drug conjugate as described in any one of items 150 to 152, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:23 and a light chain variable region containing the amino acid sequence of SEQ ID NO:24.

[0514] Item 154. The antibody-drug conjugate as described in any one of items 150 to 153, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[0515] Item 155. The antibody-drug conjugate as described in any one of items 150 to 154, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0516] Item 156. An antibody-drug conjugate having formula (I),

[0517] Ab-(LD) p (I)

[0518] in

[0519] Ab is an antibody or antigen-binding fragment that targets neoplastic cells;

[0520] D is D1;

[0521] L is the connector that covalently links Ab to D; and

[0522] p is an integer from 1 to 15.

[0523] Item 157. The antibody-drug conjugate as described in Item 156, wherein the antibody or antigen-binding fragment targets cells expressing HER2.

[0524] Item 158. The antibody-drug conjugate as described in Item 157, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

[0525] Item 159. The antibody-drug conjugate as described in Item 157 or Item 158, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[0526] Item 160. The antibody-drug conjugate as described in any one of items 157 to 159, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[0527] Item 161. The antibody-drug conjugate as described in any one of items 157 to 160, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[0528] Item 162. The antibody-drug conjugate as described in any one of items 157 to 161, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0529] Item 163. The antibody-drug conjugate as described in Item 156, wherein the antibody or antigen-binding fragment targets cells expressing CD138.

[0530] Item 164. The antibody-drug conjugate as described in Item 163, wherein the antibody or antigen-binding fragment is an anti-CD138 antibody or antigen-binding fragment.

[0531] Item 165. The antibody-drug conjugate as described in Item 163 or Item 164, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

[0532] Item 166. The antibody-drug conjugate as described in any one of items 163 to 165, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:21 and a light chain variable region containing the amino acid sequence of SEQ ID NO:22.

[0533] Item 167. The antibody-drug conjugate as described in any one of items 163 to 166, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG2a heavy chain.

[0534] Item 168. The antibody-drug conjugate as described in any one of items 163 to 167, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0535] Item 169. The antibody-drug conjugate as described in Item 156, wherein the antibody or antigen-binding fragment targets cells expressing EPHA2.

[0536] Item 170. The antibody-drug conjugate as described in Item 169, wherein the antibody or antigen-binding fragment is an anti-EPHA2 antibody or antigen-binding fragment.

[0537] Item 171. The antibody-drug conjugate as described in Item 169 or Item 170, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:16 (LCDR1), SEQ ID NO:17 (LCDR2), and SEQ ID NO:18 (LCDR3).

[0538] Item 172. The antibody-drug conjugate as described in any one of items 169 to 171, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:23 and a light chain variable region containing the amino acid sequence of SEQ ID NO:24.

[0539] Item 173. The antibody-drug conjugate as described in any one of items 169 to 172, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[0540] Item 174. The antibody-drug conjugate as described in any one of items 169 to 173, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[0541] Item 175. An antibody-drug conjugate having formula (I),

[0542] Ab-(LD) p (I)

[0543] in

[0544] Ab is an anti-HER2 antibody or antigen-binding fragment containing three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) of the amino acid sequences 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) of the amino acid sequences SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[0545] D is D2, D1, D4, D12, or D15;

[0546] L is the connector that covalently links Ab to D; and

[0547] p is an integer from 1 to 15.

[0548] Item 176. The antibody-drug conjugate as described in Item 175, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[0549] Item 177. An antibody-drug conjugate having formula (I),

[0550] Ab-(LD) p (I)

[0551] in

[0552] Ab is an anti-CD138 antibody or antigen-binding fragment containing three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) of the amino acid sequences 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) of the amino acid sequences SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

[0553] D is D2, D1, D4, D12, or D15;

[0554] L is the connector that covalently links Ab to D; and

[0555] p is an integer from 1 to 15.

[0556] Item 178. The antibody-drug conjugate as described in Item 177, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:21 and a light chain variable region containing the amino acid sequence of SEQ ID NO:22.

[0557] Item 179. An antibody-drug conjugate having formula (I),

[0558] Ab-(LD) p (I)

[0559] in

[0560] Ab is an anti-EPHA2 antibody or antigen-binding fragment containing three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) of the amino acid sequences 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) of the amino acid sequences SEQ ID NO:16 (LCDR1), SEQ ID NO:17 (LCDR2), and SEQ ID NO:18 (LCDR3).

[0561] D is D2, D1, D4, D12, or D15;

[0562] L is the connector that covalently links Ab to D; and

[0563] p is an integer from 1 to 15.

[0564] Item 180. The antibody-drug conjugate as described in Item 179, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:23 and a light chain variable region containing the amino acid sequence of SEQ ID NO:24.

[0565] Item 181. The antibody-drug conjugate as described in any one of items 156 to 180, wherein the linker is a cleavable linker.

[0566] Item 182. The antibody-drug conjugate as described in Item 181, wherein the cleavable linker comprises MC-Val-Cit-pABC.

[0567] Item 183. The antibody-drug conjugate as described in Item 181, wherein the cleavable linker comprises MC-Val-Ala-pABC.

[0568] Item 184. The antibody-drug conjugate as described in Item 181, wherein the cleavable linker comprises MC-Val-Ala-pAB.

[0569] Item 185. The antibody-drug conjugate as described in any one of items 156 to 180, wherein the linker is a non-cleavable linker.

[0570] Item 186. The antibody-drug conjugate as described in Item 185, wherein the cleavable linker comprises Mal-Hex.

[0571] Item 187. The antibody-drug conjugate as described in Item 185, wherein the cleavable linker comprises Mal-Et.

[0572] Item 188. The antibody-drug conjugate as described in Item 185, wherein the non-cleavable linker comprises Mal-Et-O-Et.

[0573] Item 189. The antibody-drug conjugate as described in any one of items 156 to 188, wherein p is 1 to 10.

[0574] Item 190. An antibody-drug conjugate as described in any one of items 156 to 189, wherein p is 2 to 8.

[0575] Item 191. The antibody-drug conjugate as described in any one of items 156 to 190, wherein p is 4 to 8.

[0576] Item 192. The antibody-drug conjugate as described in any one of items 156 to 191, wherein p is 4.

[0577] Item 193. An antibody-drug conjugate as described in any one of items 156 to 191, wherein p is 8.

[0578] Item 194. A composition comprising a plurality of copies of an antibody-drug conjugate as described in any one of items 1 to 193, wherein the average p of the antibody-drug conjugate in the composition is from about 3.5 to about 5.5.

[0579] Item 195. The composition as described in Item 194, wherein the average p of the antibody-drug conjugate in the composition is about 4.

[0580] Item 196. A composition comprising a plurality of copies of an antibody-drug conjugate as described in any one of items 1 to 193, wherein the average p of the antibody-drug conjugate in the composition is about 7 to about 9.

[0581] Item 197. The composition as described in Item 196, wherein the average p of the antibody-drug conjugate in the composition is about 8.

[0582] Item 198. A method of treating a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate as described in any one of items 1 to 193 or a composition as described in any one of items 194 to 197.

[0583] Item 199. The method as described in Item 198, wherein the proliferative condition is a hematologic malignancy or a solid tumor.

[0584] Item 200. The method described in Item 198 or Item 199, wherein the proliferative condition is a hematologic malignancy.

[0585] Item 201. The method as described in Item 199 or Item 200, wherein the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma.

[0586] Item 202. The method as described in Item 198 or Item 199, wherein the proliferative condition is a solid tumor.

[0587] Item 203. The method as described in Item 199 or Item 202, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney cancer, colorectal cancer, and esophageal cancer.

[0588] Item 204. The method of any one of items 198 to 203, wherein treatment with the antibody-drug conjugate or composition induces bystander killing of vegetative cells that do not express the target antigen but are adjacent to vegetative cells that express the target antigen.

[0589] Item 205. The method of any one of items 198 to 204, wherein the subject has one or more proliferative cells expressing the target antigen.

[0590] Item 206. The method as described in Item 205, wherein the target antigen is HER2.

[0591] Item 207. The method of Item 206, wherein the one or more proliferative cells are derived from breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary gland duct carcinoma expressing HER2.

[0592] Item 208. The method as described in Item 207, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

[0593] Item 209. The method of any one of items 206 to 208, wherein the subject is unresponsive or poorly responsive to treatment with (a) an anti-HER2 antibody administered alone and / or (b) a splicing modulator administered alone.

[0594] Item 210. The method of any one of items 206 to 209, wherein the subject is intolerant, unresponsive, or poorly responsive to treatment with the splice modulator when administered alone.

[0595] Item 211. The method as described in Item 205, wherein the target antigen is CD138.

[0596] Item 212. The method of Item 211, wherein the one or more proliferative cells are derived from multiple myeloma expressing CD138.

[0597] Item 213. The method as described in Item 211 or Item 212, wherein the subject is unresponsive or poorly responsive to treatment with (a) an anti-CD138 antibody administered alone and / or (b) a splice modulator administered alone.

[0598] Item 214. The method of any one of items 211 to 213, wherein the subject is intolerant, unresponsive, or poorly responsive to treatment with the splice modulator when administered alone.

[0599] Item 215. The method as described in Item 205, wherein the target antigen is EPHA2.

[0600] Item 216. The method of Item 215, wherein the one or more proliferative cells are derived from breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2.

[0601] Item 217. The method as described in Item 215 or Item 216, wherein the subject is unresponsive or poorly responsive to treatment with (a) an anti-EPHA2 antibody administered alone and / or (b) a splice modulator administered alone.

[0602] Item 218. The method of any one of items 215 to 217, wherein the subject is intolerant, unresponsive, or poorly responsive to treatment with the splice modulator when administered alone.

[0603] Item 219. A method for reducing or inhibiting tumor growth in a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate as described in any one of items 1 to 193 or a composition as described in any one of items 194 to 197.

[0604] Item 220. The method of Item 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 express the target antigen.

[0605] Item 221. The method as described in Item 219 or Item 220, wherein the tumor comprises one or more proliferative cells expressing the target antigen.

[0606] Item 222. The method as described in Item 221, wherein the target antigen is HER2.

[0607] Item 223. The method of Item 222, wherein the one or more proliferative cells are derived from breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary gland duct carcinoma expressing HER2.

[0608] Item 224. The method as described in Item 223, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

[0609] Item 225. The method of any one of items 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 splice modulator when administered alone.

[0610] Item 226. The method as described in Item 221, wherein the target antigen is CD138.

[0611] Item 227. The method of Item 226, wherein the one or more proliferative cells are derived from multiple myeloma expressing CD138.

[0612] Item 228. The method as described in Item 226 or Item 227, wherein the tumor is resistant or refractory to treatment with (a) an anti-CD138 antibody when administered alone and / or (b) a splice modulator when administered alone.

[0613] Item 229. The method as described in Item 221, wherein the target antigen is EPHA2.

[0614] Item 230. The method of Item 229, wherein the one or more proliferative cells are derived from breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2.

[0615] Item 231. The method as described in Item 229 or Item 230, wherein the tumor is resistant or refractory to treatment with (a) an anti-EPHA2 antibody when administered alone and / or (b) a splice modulator when administered alone.

[0616] Item 232. A method for determining whether a subject with or suspected of having a neoplastic condition will respond to treatment with an antibody-drug conjugate as described in any one of items 1 to 193 or a composition as described in any one of items 194 to 197, the method comprising providing a biological sample from the subject and contacting the biological sample with an antibody-drug conjugate as described in any one of items 1 to 193 or a composition as described in any one of items 194 to 197.

[0617] Item 233. The method as described in Item 232, wherein the biological sample is a tumor sample.

[0618] Item 234. The method as described in Item 233, wherein the tumor sample is a tumor biopsy or a blood sample.

[0619] Item 235. The method of Item 234, wherein the blood sample is selected from blood, blood fractions, or cells obtained from the blood or blood fractions.

[0620] Item 236. The method of any one of items 232 to 235, wherein the subject has one or more proliferative cells expressing the target antigen.

[0621] Item 237. The method as described in Item 236, wherein the target antigen is HER2.

[0622] Item 238. The method of Item 237, wherein the one or more proliferative cells are derived from breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary gland duct carcinoma expressing HER2.

[0623] Item 239. The method as described in Item 238, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

[0624] Item 240. The method as described in Item 236, wherein the target antigen is CD138.

[0625] Item 241. The method of Item 240, wherein the one or more proliferative cells are derived from multiple myeloma expressing CD138.

[0626] Item 242. The method as described in Item 236, wherein the target antigen is EPHA2.

[0627] Item 243. The method of Item 242, wherein the one or more proliferative cells are derived from breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2.

[0628] Item 244. An antibody-drug conjugate as described in any one of items 1 to 193 or a composition as described in any one of items 194 to 197, for the treatment of adenoid diseases.

[0629] Item 245. An antibody-drug conjugate for use as described in Item 244, wherein the hyperplastic condition is characterized by hyperplastic cells having one or more expressing target antigens.

[0630] Item 246. An antibody-drug conjugate for use as described in Item 245, wherein the target antigen is HER2.

[0631] Item 247. An antibody-drug conjugate for use as described in Item 246, wherein the neoplastic condition is HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary gland duct carcinoma.

[0632] Item 248. The antibody-drug conjugate as described in Item 247, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

[0633] Item 249. An antibody-drug conjugate for use as described in Item 245, wherein the target antigen is CD138.

[0634] Item 250. An antibody-drug conjugate for use as described in Item 249, wherein the proliferative condition is multiple myeloma expressing CD138.

[0635] Item 251. An antibody-drug conjugate for use as described in Item 245, wherein the target antigen is EPHA2.

[0636] Item 252. An antibody-drug conjugate for use as described in Item 251, wherein the proliferative disease is breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2.

[0637] Item 253. Use of the antibody-drug conjugate as described in any one of items 1 to 193 or the composition as described in any one of items 194 to 197 in the treatment of aplastic conditions.

[0638] Item 254. The use as described in Item 253, wherein the hyperplastic condition is characterized by hyperplastic cells having one or more expressing target antigens.

[0639] Item 255. Use as described in Item 254, wherein the target antigen is HER2.

[0640] Item 256. Use as described in Item 255, wherein the neoplastic condition is HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary gland duct carcinoma.

[0641] Item 257. The use as described in Item 256, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

[0642] Item 258. Use as described in Item 254, wherein the target antigen is CD138.

[0643] Item 259. Use as described in Item 258, wherein the proliferative condition is CD138-expressing multiple myeloma.

[0644] Item 260. Use as described in Item 254, wherein the target antigen is EPHA2.

[0645] Item 261. The use as described in Item 260, wherein the proliferative condition is breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2.

[0646] Item 262. Use of the antibody-drug conjugate as described in any one of items 1 to 193 or the composition as described in any one of items 194 to 197 in a method of manufacturing a medicament for treating aplastic conditions.

[0647] Item 263. The use as described in Item 262, wherein the hyperplastic condition is characterized by hyperplastic cells having one or more expressing target antigens.

[0648] Item 264. Use as described in Item 263, wherein the target antigen is HER2.

[0649] Item 265. Use as described in Item 264, wherein the neoplastic condition is HER2-expressing breast cancer, ovarian cancer, gastric cancer, lung cancer, uterine cancer, osteosarcoma, or salivary gland duct carcinoma.

[0650] Item 266. The use as described in Item 265, wherein the lung cancer is lung adenocarcinoma and / or the uterine cancer is serous endometrial carcinoma.

[0651] Item 267. Use as described in Item 263, wherein the target antigen is CD138.

[0652] Item 268. Use as described in Item 267, wherein the proliferative condition is CD138-expressing multiple myeloma.

[0653] Item 269. Use as described in Item 263, wherein the target antigen is EPHA2.

[0654] Item 270. Use as described in Item 269, wherein the proliferative condition is breast cancer, prostate cancer, ovarian cancer, lung cancer, melanoma, colon cancer, or esophageal cancer expressing EPHA2.

[0655] Item 271. A pharmaceutical composition comprising an antibody-drug conjugate as described in any one of items 1 to 193 or a composition as described in any one of items 194 to 197, and a pharmaceutically acceptable carrier.

[0656] Item 272. A method for producing an antibody-drug conjugate as described in any one of items 1 to 193, the method comprising reacting an antibody or antigen-binding fragment with a linker conjugated to a splice modulator under conditions permissible for conjugation.

[0657] Item 273. A compound having formula (III):

[0658] or a pharmaceutically acceptable salt thereof, wherein:

[0659] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0660] R 2 It may not exist or may be a connector;

[0661] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[0662] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0663] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[0664] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and -C(=O)-OR 17;

[0665] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0666] Z is selected from

[0667] Where R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[0668] Where R 6 and R 7 At least one of them is hydrogen; and

[0669] Where R 2 If it is a connector, then R 6 and R 7 None of them are connectors, and if R 6 Or R 7 If it is a connector, then R 2 Does not exist.

[0670] Item 274. A compound having formula (V):

[0671] or a pharmaceutically acceptable salt thereof, wherein:

[0672] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0673] R 2 It may not exist or may be a connector;

[0674] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[0675] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0676] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[0677] R 15 and R 16 Each is independently selected from hydrogen, R 17 -C(=O)-R 17 and -C(=O)-OR 17 ;and

[0678] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0679] Where R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[0680] Where R 6 and R 7 At least one of them is hydrogen; and

[0681] Where R 2 If it is a connector, then R 6 and R 7 None of them are connectors, and if R 6 Or R 7 If it is a connector, then R 2 Does not exist.

[0682] Item 275. A compound having formula (VII):

[0683] or a pharmaceutically acceptable salt thereof, wherein:

[0684] R 1 and R 9 Each is independently selected from the absence of, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0685] R 2 It may not exist or may be a connector;

[0686] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[0687] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0688] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[0689] R 10 Selected from hydrogen, C1-C6 alkyl groups, -C(=O)-(C1-C6 alkyl) groups, and -CD3;

[0690] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and -C(=O)-OR 17 ;

[0691] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0692] a is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0693] where 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 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[0694] where R 6 and R 7 At least one of them is hydrogen;

[0695] Where R 2 If it is a connector, then R 6 and R 7 None of them are connectors, and if R 6 or R 7 If it is a connector, then R 2 It does not exist; and

[0696] where R 1 and R 9 They cannot both be absent at the same time.

[0697] Item 276. A compound having formula (IX):

[0698] or a pharmaceutically acceptable salt thereof, wherein:

[0699] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[0700] R 2 It's a connector;

[0701] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[0702] R 4 Selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[0703] R 10 Selected from 3- to 10-membered carbon rings and 3- to 10-membered heterocycles, each of which is composed of 0 to 3 R... a Replace, where each R a Independently selected from halogens, C1-C6 alkyl groups, -O-(C1-C6)alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylhydroxy groups, and -S (=O). w -(4- to 7-membered heterocycles), 4- to 7-membered carbon rings and 4- to 7-membered heterocycles;

[0704] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and -C(=O)-OR 17 ;and

[0705] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[0706] where R 1 R 2 R3 R 4 R 10 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups; and

[0707] Each R a Independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, -NR 15 R 16 C1-C6 alkyl groups, -(C=O)-(C1-C6 alkyl) groups, -(C=O)-(C1-C6 alkyl)-(C3-C 10 Heterocyclic groups, -S (=O) w -(C3-C8 heterocyclic groups and C1-C6 alkylcarboxylic acid groups, each substituted by 0, 1 or 2 independently selected groups from: halogen, hydroxyl group, -NR 15 R 16 and C1-C3 alkyl groups.

[0708] Item 277. A compound selected from those having the formula

[0709] Compounds and their pharmaceutically acceptable salts.

[0710] Item 278. A pharmaceutical composition comprising a compound as described in Item 277 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0711] Item 279. A compound selected from those having the formula

[0712] Compounds and their pharmaceutically acceptable salts.

[0713] Item 280. A pharmaceutical composition comprising a compound as described in Item 279 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0714] Item 281. A compound selected from those having the formula

[0715] Compounds and their pharmaceutically acceptable salts.

[0716] Item 282. A pharmaceutical composition comprising a compound as described in Item 281 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0717] Item 283. A compound selected from those having the formula

[0718] Compounds and their pharmaceutically acceptable salts.

[0719] Item 284. A pharmaceutical composition comprising a compound as described in Item 283 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0720] Item 285. A compound selected from those having the formula

[0721] Compounds and their pharmaceutically acceptable salts.

[0722] Item 286. A pharmaceutical composition comprising a compound as described in Item 285 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0723] Item 287. A compound selected from those having the formula

[0724] Compounds and their pharmaceutically acceptable salts.

[0725] Item 288. A pharmaceutical composition comprising a compound as described in Item 287 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0726] Item 289. A compound selected from those having the formula

[0727] Compounds and their pharmaceutically acceptable salts.

[0728] Item 290. A pharmaceutical composition comprising a compound as described in Item 289 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0729] Item 291. A compound selected from those having the formula

[0730] Compounds and their pharmaceutically acceptable salts.

[0731] Item 292. A pharmaceutical composition comprising a compound as described in Item 291 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0732] Item 293. A compound selected from those having the formula

[0733] Compounds and their pharmaceutically acceptable salts.

[0734] Item 294. A pharmaceutical composition comprising a compound as described in Item 293 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0735] Item 295. A compound selected from those having the formula

[0736] Compounds and their pharmaceutically acceptable salts.

[0737] Item 296. A pharmaceutical composition comprising a compound as described in Item 295 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0738] Item 297. A compound selected from those having the formula

[0739] Compounds and their pharmaceutically acceptable salts.

[0740] Item 298. A pharmaceutical composition comprising a compound as described in Item 297 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0741] Item 299. A compound selected from those having the formula

[0742] Compounds and their pharmaceutically acceptable salts.

[0743] Item 300. A pharmaceutical composition comprising a compound as described in Item 299 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0744] Item 301. A method of treating a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition as described in any one of items 273 to 300.

[0745] Item 302. A method for reducing or inhibiting tumor growth in a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition as described in any one of items 273 to 300.

[0746] Item 303. A method for inducing at least one neoantigen, the method comprising contacting proliferative cells with an effective amount of a splicing regulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197, thereby inducing the production of at least one neoantigen.

[0747] Item 304. The method of item 303, wherein the at least one neoantigen comprises the amino acid sequence of any one of SEQ ID NO:37-65.

[0748] Item 305. The method of item 304, wherein the at least one neoantigen comprises the amino acid sequence of SEQ ID NO:37.

[0749] Item 306. The method of item 304, wherein the at least one neoantigen comprises the amino acid sequence of SEQ ID NO:39.

[0750] Item 307. The method of item 304, wherein the at least one neoantigen comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[0751] Item 308. The method of any one of items 303 to 307, wherein the proliferative cells are present in an in vitro cell culture.

[0752] Item 309. The method of Item 308, wherein the proliferative cells are obtained from the subject.

[0753] Item 310. The method of any one of items 303 to 307, wherein the proliferative cells are present in the subject.

[0754] Item 311. The method of any one of items 303 to 310, wherein the proliferative cells are derived from hematologic malignancies or solid tumors.

[0755] Item 312. The method of Item 311, wherein the hematologic malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

[0756] Item 313. The method as described in Item 311 or Item 312, wherein the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma.

[0757] Item 314. The method of Item 311, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary gland duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney cancer, colorectal cancer, and esophageal cancer.

[0758] Item 315. A method for inducing at least one neoantigen and / or T cell response in a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197.

[0759] Item 316. A method of treating a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject an effective amount of a splice modulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197, wherein administration of the splice modulator, antibody-drug conjugate, or composition induces at least one neoantigen and / or T cell response.

[0760] Item 317. The method of Item 315 or Item 316, wherein the at least one neoantigen comprises the amino acid sequence of any one of SEQ ID NO:37-65.

[0761] Item 318. The method of item 317, wherein the at least one neoantigen comprises the amino acid sequence of SEQ ID NO:37.

[0762] Item 319. The method of Item 317, wherein the at least one neoantigen comprises the amino acid sequence of SEQ ID NO:39.

[0763] Item 320. The method of Item 317, wherein the at least one neoantigen comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[0764] Item 321. The method of any one of items 315 to 320, wherein the amount of splice modulator, antibody-drug conjugate, or composition administered is reduced due to the induction of at least one neoantigen and / or T cell response compared to a standard dose of the splice modulator, antibody-drug conjugate, or composition.

[0765] Item 322. The method of any one of items 315 to 321, wherein the amount of the splice modulator, antibody-drug conjugate, or composition administered is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to a standard dose of the splice modulator, antibody-drug conjugate, or composition.

[0766] Item 323. The method of any one of items 315 to 322, wherein the splice modulator, antibody-drug conjugate, or composition is administered at a frequency at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% lower than the standard dosing regimen of the splice modulator, antibody-drug conjugate, or composition.

[0767] Item 324. The method of any one of items 315 to 323, wherein the amount and / or dose of the splice modulator, antibody-drug conjugate, or composition administered results in lower systemic toxicity and / or improved tolerability.

[0768] Item 325. The method of any one of items 315 to 324, further comprising administering at least one additional therapy.

[0769] Item 326. The method of Item 325, wherein the amount of splice modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy administered is reduced due to the induction of at least one neoantigen and / or T cell response compared to a standard dose of the splice modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy.

[0770] Item 327. The method as described in Item 325 or Item 326, wherein the amount of the splice 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% compared to the standard dose of the splice modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy.

[0771] Item 328. The method of any one of items 325 to 327, wherein the frequency of administration of the splice modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy is at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% lower than the standard dosing regimen of the splice modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy.

[0772] Item 329. The method of any one of items 325 to 328, wherein the amount and / or dose of the splice modulator, antibody-drug conjugate, composition, and / or the at least one additional therapy results in lower systemic toxicity and / or improved tolerability.

[0773] Item 330. The method of any one of items 325 to 329, wherein the application of the splice modulator, antibody-drug conjugate, or composition is initiated prior to the application of the at least one additional therapy.

[0774] Item 331. The method of any one of items 325 to 329, wherein the application of the splice modulator, antibody-drug conjugate, or composition is initiated after the administration of the at least one additional therapy.

[0775] Item 332. The method of any one of items 325 to 329, wherein the administration of the splice modulator, antibody-drug conjugate, or composition is initiated simultaneously with the administration of the at least one additional therapy.

[0776] Item 333. The method of any one of items 315 to 332, wherein the splice modulator, antibody-drug conjugate, or composition is applied at least once after the initial application.

[0777] Item 334. The method of Item 333, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced compared to the amount used for initial administration.

[0778] Item 335. The method of Item 333 or Item 334, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced compared to a standard dose of the splice modulator, antibody-drug conjugate, or composition.

[0779] Item 336. The method of any one of items 333 to 335, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated application is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to a standard dose of the splice modulator, antibody-drug conjugate, or composition.

[0780] Item 337. The method of any one of items 325 to 336, wherein the at least one additional therapy is repeated at least once after the initial application.

[0781] Item 338. The method of Item 337, wherein the amount of the at least one additional therapy used for repeated administration is reduced compared to the amount used for the initial administration.

[0782] Item 339. The method as described in Item 337 or Item 338, wherein the amount of the at least one additional therapy used for repeated administration is reduced compared to the standard dose of the at least one additional therapy.

[0783] Item 340. The method of any one of items 337 to 339, wherein the amount of the at least one additional therapy used for repeated administration is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dose of the at least one additional therapy.

[0784] Item 341. The method of any one of items 337 to 340, wherein the repeated administration of the splice modulator, antibody-drug conjugate, or composition is performed concurrently with the repeated administration of the at least one additional therapy.

[0785] Item 342. The method of any one of items 337 to 340, wherein the repeated administration of the splice modulator, antibody-drug conjugate, or composition is performed sequentially or separately from the repeated administration of the at least one additional therapy.

[0786] Item 343. The method of any one of items 325 to 342, wherein the at least one additional therapy includes the administration of a checkpoint inhibitor.

[0787] Item 344. The method as described in Item 343, wherein the subject is intolerant, unresponsive, or poorly responsive to the checkpoint inhibitor when administered alone.

[0788] Item 345. The method as described in Item 343 or Item 344, wherein the checkpoint inhibitor targets CTLA4, PD1, PDL1, OX40, CD40, GITR, LAG3, TIM3 and / or KIR.

[0789] Item 346. The method of any one of items 343 to 345, wherein the checkpoint inhibitor targets CTLA4, OX40, CD40 and / or GITR.

[0790] Item 347. The method of any one of items 343 to 346, wherein the checkpoint inhibitor comprises a cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) pathway inhibitor.

[0791] Item 348. The method as described in Item 347, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

[0792] Item 349. The method as described in Item 348, wherein the anti-CTLA4 antibody is ipilimumab.

[0793] Item 350. The method of any one of items 343 to 346, wherein the checkpoint inhibitor comprises a programmed death-1 pathway (PD1) inhibitor.

[0794] Item 351. The method of Item 350, wherein the PD1 inhibitor is an anti-PD1 antibody.

[0795] Item 352. The method as described in Item 351, wherein the anti-PD1 antibody is nivolumab.

[0796] Item 353. The method of Item 350, wherein the PD1 inhibitor is an anti-PDL1 antibody.

[0797] Item 354. The method as described in Item 353, wherein the anti-PDL1 antibody is atezolizumab.

[0798] Item 355. The method of any one of items 343 to 346, wherein the checkpoint inhibitor comprises a CTLA4 inhibitor and a PD1 inhibitor.

[0799] Item 356. The method as described in Item 355, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

[0800] Item 357. The method as described in Item 356, wherein the anti-CTLA4 antibody is ipilimumab.

[0801] Item 358. The method of Item 355, wherein the PD1 inhibitor is an anti-PD1 antibody.

[0802] Item 359. The method as described in Item 358, wherein the anti-PD1 antibody is nivolumab.

[0803] Item 360. The method of Item 355, wherein the PD1 inhibitor is an anti-PDL1 antibody.

[0804] Item 361. The method of Item 360, wherein the anti-PDL1 antibody is atezolizumab.

[0805] Item 362. The method of any one of items 325 to 342, wherein the at least one additional therapy includes administration of a neoantigen vaccine.

[0806] Item 363. The method of Item 362, wherein the splicing modulator, antibody-drug conjugate, or composition is administered prior to the administration of the neoantigen vaccine.

[0807] Item 364. The method of Item 362, wherein the splicing modulator, antibody-drug conjugate, or composition is administered after administration of the neoantigen vaccine.

[0808] Item 365. The method of Item 362, wherein the splicing modulator, antibody-drug conjugate, or composition is administered concurrently with the administration of the neoantigen vaccine.

[0809] Item 366. The method of any one of items 362 to 365, wherein the splice modulator, antibody-drug conjugate, or composition is applied at least once after the initial application.

[0810] Item 367. The method of Item 366, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced compared to the amount used for initial administration.

[0811] Item 368. The method of any one of items 362 to 367, wherein the neoantigen vaccine comprises at least one neoantigen peptide.

[0812] Item 369. The method of Item 368, wherein the length of the at least one neoantigen peptide is in the range of about 10 to about 50 or about 10 to about 35 amino acids.

[0813] Item 370. The method of Item 368 or Item 369, wherein the length of the at least one neoantigen peptide is in the range of about 15 to about 25 amino acids.

[0814] Item 371. The method of any one of items 368 to 370, wherein the at least one neoantigen peptide comprises one or more neoantigen sequences.

[0815] Item 372. The method of Item 371, wherein the neoantigen sequence is a neoantigen sequence specific to the subject.

[0816] Item 373. The method as described in Item 371, wherein the neoantigen sequence is a universal neoantigen sequence.

[0817] Item 374. The method of any one of items 371 to 373, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO: 37-65.

[0818] Item 375. The method of Item 374, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:37.

[0819] Item 376. The method of Item 374, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:39.

[0820] Item 377. The method of Item 374, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[0821] Item 378. The method of any one of items 371 to 373, wherein the novel antigen sequence comprises the amino acid sequence of any one of SEQ ID NO:66-93 or the antigen portion of any one of SEQ ID NO:66-93.

[0822] Item 379. The method of Item 378, wherein the novel antigen sequence comprises the amino acid sequence of SEQ ID NO:66 or the antigen portion of SEQ ID NO:66.

[0823] Item 380. The method of Item 378, wherein the novel antigen sequence comprises the amino acid sequence of any one of SEQ ID NO:74-77, or the antigen portion of any one of SEQ ID NO:74-77.

[0824] Item 381. The method of any one of items 374 to 380, wherein the length of the neoantigen sequence and / or antigenic moiety is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[0825] Item 382. The method of any one of items 374 to 381, wherein the neoantigen sequence and / or antigen portion is a personalized neoantigen vaccine for the subject.

[0826] Item 383. The method of Item 382, ​​wherein the neoantigen sequence and / or antigenic portion has been identified by sequencing at least one neoantigen induced in the subject by administration of an effective amount of the splicing modulator, antibody-drug conjugate, or composition.

[0827] Item 384. The method as described in Item 382 or Item 383, wherein the neoantigen sequence and / or antigenic portion is capable of binding to at least one HLA allele expressed in the subject.

[0828] Item 385. The method of any one of items 374 to 381, wherein the neoantigen sequence and / or antigen portion is a universal neoantigen vaccine.

[0829] Item 386. The method of Item 385, wherein the neoantigen 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 subject population suffering from the neoplastic condition.

[0830] Item 387. The method of Item 385 or Item 386, wherein the neoantigen sequence and / or antigen portion is capable of eliciting a T-cell response against tumors present in at least 1%, at least 5%, or at least 10% of the subject population suffering from the neoplastic condition.

[0831] Item 388. The method of any one of items 385 to 387, wherein the at least one neoantigen peptide comprises a neoantigen sequence induced by contacting the proliferating cells with an effective amount of the splicing regulator, antibody-drug conjugate, or composition.

[0832] Item 389. The method of any one of items 362 to 367, wherein the neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable carrier.

[0833] Item 390. The method of Item 389, wherein the at least one neoantigen peptide is linked to the pharmaceutically acceptable carrier.

[0834] Item 391. The method as described in Item 389 or Item 390, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

[0835] Item 392. The method of any one of items 389 to 391, wherein the neoantigen peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

[0836] Item 393. The method of any one of items 389 to 391, wherein the neoantigen peptide and the pharmaceutically acceptable vector are expressed as a fusion protein.

[0837] Item 394. The method of any one of items 362 to 367, wherein the neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable diluent.

[0838] Item 395. The method of any one of items 362 to 367, wherein the neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable adjuvant.

[0839] Item 396. The method of Item 388, wherein the proliferative cells are present in an in vitro cell culture.

[0840] Item 397. The method as described in Item 396, wherein the proliferative cells are obtained from the subject.

[0841] Item 398. The method of Item 388, wherein the proliferative cell is present in the subject.

[0842] Item 399. The method of any one of items 362 to 367, wherein the neoantigen vaccine comprises at least one neoantigen mRNA.

[0843] Item 400. The method of Item 399, wherein the at least one neoantigen mRNA encodes one or more neoantigen sequences.

[0844] Item 401. The method of item 400, wherein the neoantigen sequence is a neoantigen sequence specific to the subject.

[0845] Item 402. The method of item 400, wherein the neoantigen sequence is a universal neoantigen sequence.

[0846] Item 403. The method of any one of items 400 to 402, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:37-65.

[0847] Item 404. The method of Item 403, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:37.

[0848] Item 405. The method of item 403, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:39.

[0849] Item 406. The method of Item 403, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[0850] Item 407. The method of any one of items 400 to 402, wherein the novel antigen sequence comprises the amino acid sequence of any one of SEQ ID NO:66-93 or the antigen portion of any one of SEQ ID NO:66-93.

[0851] Item 408. The method of Item 407, wherein the novel antigen sequence comprises the amino acid sequence of SEQ ID NO:66 or the antigen portion of SEQ ID NO:66.

[0852] Item 409. The method of Item 407, wherein the novel antigen sequence comprises the amino acid sequence of any one of SEQ ID NO:74-77, or the antigen portion of any one of SEQ ID NO:74-77.

[0853] Item 410. The method of any one of items 403 to 409, wherein the length of the neoantigen sequence and / or antigenic portion is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[0854] Item 411. The method of any one of items 403 to 410, wherein the neoantigen sequence and / or antigen portion is a personalized neoantigen vaccine for the subject.

[0855] Item 412. The method of Item 411, wherein the neoantigen sequence and / or antigenic portion has been identified by sequencing at least one neoantigen induced in the subject by administration of an effective amount of the splicing modulator, antibody-drug conjugate, or composition.

[0856] Item 413. The method as described in Item 411 or Item 412, wherein the neoantigen sequence and / or antigenic portion is capable of binding to at least one HLA allele expressed in the subject.

[0857] Item 414. The method of any one of items 403 to 410, wherein the neoantigen sequence and / or antigen portion is a universal neoantigen vaccine.

[0858] Item 415. The method of Item 414, wherein the neoantigen 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 subject population suffering from the neoplastic condition.

[0859] Item 416. The method of Item 414 or Item 415, wherein the neoantigen sequence and / or antigen portion is capable of eliciting a T-cell response against tumors present in at least 1%, at least 5%, or at least 10% of the subject population suffering from the neoplastic condition.

[0860] Item 417. The method of any one of items 414 to 416, wherein the at least one neoantigen mRNA encodes a neoantigen sequence induced by contacting a proliferative cell with an effective amount of the splicing regulator, antibody-drug conjugate, or composition.

[0861] Item 418. The method of any one of items 362 to 367, wherein the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable vector.

[0862] Item 419. The method of Item 418, wherein the at least one neoantigen mRNA is ligated to the pharmaceutically acceptable vector.

[0863] Item 420. The method as described in Item 418 or Item 419, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

[0864] Item 421. The method of any one of items 362 to 367, wherein the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable diluent.

[0865] Item 422. The method of any one of items 362 to 367, wherein the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable adjuvant.

[0866] Item 423. The method of any one of items 399 to 422, wherein the neoantigen mRNA is encapsulated by an encapsulating agent.

[0867] Item 424. The method as described in Item 423, wherein the encapsulating agent is a liposome.

[0868] Item 425. The method as described in Item 423, wherein the encapsulating agent is a nanoparticle.

[0869] Item 426. The method of Item 417, wherein the proliferative cells are present in an in vitro cell culture.

[0870] Item 427. The method as described in Item 426, wherein the proliferative cells are obtained from the subject.

[0871] Item 428. The method as described in Item 417, wherein the proliferative cell is present in the subject.

[0872] Item 429. The method of any one of items 325 to 342, wherein the at least one additional therapy comprises administration of a cytokine or a cytokine analogue.

[0873] Item 430. The method of Item 429, wherein the subject is intolerant, unresponsive, or poorly responsive to the cytokine or cytokine analogue when administered alone.

[0874] Item 431. The method as described in Item 429 or Item 430, wherein the cytokine or cytokine analog comprises a T-cell enhancer.

[0875] Item 432. The method of any one of items 429 to 431, wherein the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, IL-15, IFNγ and / or TNFα.

[0876] Item 433. The method of any one of items 325 to 342, wherein the at least one additional therapy comprises the administration of engineered tumor-targeting T cells.

[0877] Item 434. The method of any one of items 315 to 433, further comprising detecting one or more neoantigens and / or T-cell responses in the subject after administration of the splice modulator, antibody-drug conjugate, or composition.

[0878] Item 435. The method of Item 434, further comprising continuing to administer the splicing modulator, antibody-drug conjugate, or composition if one or more neoantigens and / or T-cell responses are detected.

[0879] Item 436. The method of Item 434 or Item 435, further comprising continuing to administer the splicing modulator, antibody-drug conjugate, or composition at a lower frequency and / or at a reduced dose if one or more neoantigens and / or a T-cell response is detected.

[0880] Item 437. The method of any one of items 434 to 436, wherein the detection of one or more neoantigens and / or T-cell responses in the subject indicates efficacy of treatment with the splice modulator, antibody-drug conjugate, or composition.

[0881] Item 438. The method of any one of items 315 to 437, wherein the subject has a nonsynonymous mutation burden of about 150 mutations or less.

[0882] Item 439. The method of any one of items 315 to 438, wherein the subject has a nonsynonymous mutation burden of about 100 mutations or less.

[0883] Item 440. The method of any one of items 315 to 439, wherein the subject has a nonsynonymous mutation burden of about 50 mutations or less.

[0884] Item 441. The method of any one of items 315 to 440, wherein the proliferative condition is a hematologic malignancy or a solid tumor.

[0885] Item 442. The method as described in Item 441, wherein the hematologic malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

[0886] Item 443. The method as described in Item 441 or Item 442, wherein the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma.

[0887] Item 444. The method as described in Item 441, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary gland duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney cancer, colorectal cancer, and esophageal cancer.

[0888] Item 445. A method for treating a subject who has or is suspected of having a neoplastic condition, the method comprising:

[0889] (a) The subject is given an effective amount of a splicing modulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197, wherein the administration of the splicing modulator, antibody-drug conjugate, or composition induces at least one neoantigen and / or T cell response;

[0890] (b) Detecting one or more neoantigens and / or T-cell responses in the subject after administration of the splicing modulator, antibody-drug conjugate, or composition; and

[0891] (c) If one or more new antigens and / or T cell responses are detected, continue administration of the splicing modulator, antibody-drug conjugate, or composition.

[0892] Item 446. The method of Item 445, wherein the detection of one or more neoantigens and / or T-cell responses in the subject indicates efficacy of treatment with the splice modulator, antibody-drug conjugate, or composition.

[0893] Item 447. The method as described in Item 445 or Item 446, wherein the one or more neoantigens comprise the amino acid sequence of any one of SEQ ID NO:37-65.

[0894] Item 448. The method of Item 447, wherein the one or more neoantigens comprises the amino acid sequence of SEQ ID NO:37.

[0895] Item 449. The method of Item 447, wherein the one or more neoantigens comprises the amino acid sequence of SEQ ID NO:39.

[0896] Item 450. The method of Item 447, wherein the one or more neoantigens comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[0897] Item 451. A method of treating a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197; and at least one additional therapy.

[0898] Item 452. The method of Item 451, wherein the at least one additional therapy comprises at least one, at least two, at least three, at least four, or at least five additional therapies.

[0899] Item 453. The method as described in Item 451 or Item 452, wherein administration of the splicing modulator, antibody-drug conjugate, or composition induces at least one neoantigen and / or T cell response.

[0900] Item 454. The method of any one of items 451 to 453, wherein the amount of the splice 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% compared to the standard dose of the splice modulator, antibody-drug conjugate, or composition, and / or the at least one additional therapy.

[0901] Item 455. The method of any one of items 451 to 454, wherein the frequency of administration of the splice modulator, antibody-drug conjugate, or composition, and / or the at least one additional therapy is at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% lower than the standard dosing regimen of the splice modulator, antibody-drug conjugate, or composition, and / or the at least one additional therapy.

[0902] Item 456. The method of any one of items 451 to 455, wherein the amount and / or dose of the splice modulator, antibody-drug conjugate, or composition, and / or the at least one additional therapy results in lower systemic toxicity and / or improved tolerability.

[0903] Item 457. The method of any one of items 451 to 456, wherein the application of the splice modulator, antibody-drug conjugate, or composition is initiated prior to the application of the at least one additional therapy.

[0904] Item 458. The method of any one of items 451 to 456, wherein the application of the splice modulator, antibody-drug conjugate, or composition is initiated after the administration of the at least one additional therapy.

[0905] Item 459. The method of any one of items 451 to 456, wherein the administration of the splice modulator, antibody-drug conjugate, or composition is initiated simultaneously with the administration of the at least one additional therapy.

[0906] Item 460. The method of any one of items 451 to 460, wherein the splice modulator, antibody-drug conjugate, or composition is repeated at least once after the initial application.

[0907] Item 461. The method of Item 460, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced compared to the amount used for initial administration.

[0908] Item 462. The method of Item 460 or Item 461, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced compared to a standard dose of the splice modulator, antibody-drug conjugate, or composition.

[0909] Item 463. The method of any one of items 460 to 462, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to a standard dose of the splice modulator, antibody-drug conjugate, or composition.

[0910] Item 464. The method of any one of items 451 to 463, wherein the at least one additional therapy is repeated at least once after the initial application.

[0911] Item 465. The method of item 464, wherein the amount of the at least one additional therapy used for repeated administration is reduced compared to the amount used for the initial administration.

[0912] Item 466. The method as described in Item 464 or Item 465, wherein the amount of the at least one additional therapy used for repeated administration is reduced compared to the standard dose of the at least one additional therapy.

[0913] Item 467. The method of any one of items 464 to 466, wherein the amount of the at least one additional therapy used for repeated administration is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dose of the at least one additional therapy.

[0914] Item 468. The method of any one of items 464 to 467, wherein the repeated administration of the splice modulator, antibody-drug conjugate, or composition is performed concurrently with the repeated administration of the at least one additional therapy.

[0915] Item 469. The method of any one of items 464 to 467, wherein the repeated administration of the splice modulator, antibody-drug conjugate, or composition is performed sequentially or separately from the repeated administration of the at least one additional therapy.

[0916] Item 470. The method of any one of items 451 to 469, wherein the at least one additional therapy includes the administration of a checkpoint inhibitor.

[0917] Item 471. The method of Item 470, wherein the subject is intolerant, unresponsive, or poorly responsive to the checkpoint inhibitor when administered alone.

[0918] Item 472. The method as described in Item 470 or Item 471, wherein the checkpoint inhibitor targets CTLA4, PD1, PDL1, OX40, CD40, GITR, LAG3, TIM3 and / or KIR.

[0919] Item 473. The method of any one of items 470 to 472, wherein the checkpoint inhibitor targets CTLA4, OX40, CD40 and / or GITR.

[0920] Item 474. The method of any one of items 470 to 473, wherein the checkpoint inhibitor comprises a cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) pathway inhibitor.

[0921] Item 475. The method as described in Item 474, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

[0922] Item 476. The method as described in Item 475, wherein the anti-CTLA4 antibody is ipilimumab.

[0923] Item 477. The method of any one of items 470 to 473, wherein the checkpoint inhibitor comprises a programmed death-1 pathway (PD1) inhibitor.

[0924] Item 478. The method as described in Item 477, wherein the PD1 inhibitor is an anti-PD1 antibody.

[0925] Item 479. The method as described in Item 478, wherein the anti-PD1 antibody is nivolumab.

[0926] Item 480. The method of Item 477, wherein the PD1 inhibitor is an anti-PDL1 antibody.

[0927] Item 481. The method as described in Item 480, wherein the anti-PDL1 antibody is atezolizumab.

[0928] Item 482. The method of any one of items 470 to 473, wherein the checkpoint inhibitor comprises a CTLA4 inhibitor and a PD1 inhibitor.

[0929] Item 483. The method as described in Item 482, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

[0930] Item 484. The method as described in Item 483, wherein the anti-CTLA4 antibody is ipilimumab.

[0931] Item 485. The method of Item 482, wherein the PD1 inhibitor is an anti-PD1 antibody.

[0932] Item 486. The method as described in Item 485, wherein the anti-PD1 antibody is nivolumab.

[0933] Item 487. The method of Item 482, wherein the PD1 inhibitor is an anti-PDL1 antibody.

[0934] Item 488. The method as described in Item 487, wherein the anti-PDL1 antibody is atezolizumab.

[0935] Item 489. The method of any one of items 451 to 469, wherein the at least one additional therapy includes administration of a neoantigen vaccine.

[0936] Item 490. The method of Item 489, wherein the splicing modulator, antibody-drug conjugate, or composition is administered prior to the administration of the neoantigen vaccine.

[0937] Item 491. The method of Item 489, wherein the splicing modulator, antibody-drug conjugate, or composition is administered after administration of the neoantigen vaccine.

[0938] Item 492. The method of Item 489, wherein the splicing modulator, antibody-drug conjugate, or composition is administered concurrently with the administration of the neoantigen vaccine.

[0939] Item 493. The method of any one of items 489 to 492, wherein the splice modulator, antibody-drug conjugate, or composition is repeated at least once after the initial application.

[0940] Item 494. The method of Item 493, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced compared to the amount used for initial administration.

[0941] Item 495. The method of any one of items 489 to 494, wherein the neoantigen vaccine comprises at least one neoantigen peptide.

[0942] Item 496. The method of Item 495, wherein the length of the at least one neoantigen peptide is in the range of about 10 to about 50 amino acids or about 10 to about 35 amino acids.

[0943] Item 497. The method as described in Item 495 or Item 496, wherein the length of the at least one neoantigen peptide is in the range of about 15 to about 25 amino acids.

[0944] Item 498. The method of any one of items 495 to 497, wherein the at least one neoantigen peptide comprises one or more neoantigen sequences.

[0945] Item 499. The method of Item 498, wherein the neoantigen sequence is a neoantigen sequence specific to the subject.

[0946] Item 500. The method as described in Item 498, wherein the neoantigen sequence is a universal neoantigen sequence.

[0947] Item 501. The method of any one of items 498 to 500, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:37-65.

[0948] Item 502. The method of Item 501, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:37.

[0949] Item 503. The method of item 501, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:39.

[0950] Item 504. The method of Item 501, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[0951] Item 505. The method of any one of items 498 to 500, wherein the novel antigen sequence comprises the amino acid sequence of any one of SEQ ID NO:66-93 or the antigen portion of any one of SEQ ID NO:66-93.

[0952] Item 506. The method of Item 505, wherein the novel antigen sequence comprises the amino acid sequence of SEQ ID NO:66 or the antigen portion of SEQ ID NO:66.

[0953] Item 507. The method of Item 505, wherein the novel antigen sequence comprises the amino acid sequence of any one of SEQ ID NO:74-77, or the antigen portion of any one of SEQ ID NO:74-77.

[0954] Item 508. The method of any one of items 501 to 507, wherein the length of the neoantigen sequence and / or antigenic portion is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[0955] Item 509. The method of any one of items 501 to 508, wherein the neoantigen sequence and / or antigen portion is a personalized neoantigen vaccine for the subject.

[0956] Item 510. The method of Item 509, wherein the neoantigen sequence and / or antigenic portion has been identified by sequencing at least one neoantigen induced in the subject by administration of an effective amount of the splicing modulator, antibody-drug conjugate, or composition.

[0957] Item 511. The method of Item 509 or Item 510, wherein the neoantigen sequence and / or antigenic portion is capable of binding to at least one HLA allele expressed in the subject.

[0958] Item 512. The method of any one of items 501 to 508, wherein the neoantigen sequence and / or antigen portion is a universal neoantigen vaccine.

[0959] Item 513. The method of Item 512, wherein the neoantigen 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 subject population suffering from the neoplastic condition.

[0960] Item 514. The method of Item 512 or Item 513, wherein the neoantigen sequence and / or antigen portion is capable of eliciting a T-cell response against tumors present in at least 1%, at least 5%, or at least 10% of the subject population suffering from the neoplastic condition.

[0961] Item 515. The method of any one of items 512 to 514, wherein the at least one neoantigen peptide comprises a neoantigen sequence induced by contacting the proliferating cells with an effective amount of the splicing regulator, antibody-drug conjugate, or composition.

[0962] Item 516. The method of any one of items 489 to 494, wherein the neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable carrier.

[0963] Item 517. The method of Item 516, wherein the at least one neoantigen peptide is linked to the pharmaceutically acceptable carrier.

[0964] Item 518. The method as described in Item 516 or Item 517, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

[0965] Item 519. The method of any one of items 516 to 518, wherein the neoantigen peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

[0966] Item 520. The method of any one of items 516 to 518, wherein the neoantigen peptide and the pharmaceutically acceptable vector are expressed as a fusion protein.

[0967] Item 521. The method of any one of items 489 to 494, wherein the neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable diluent.

[0968] Item 522. The method of any one of items 489 to 494, wherein the neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable adjuvant.

[0969] Item 523. The method of Item 515, wherein the proliferative cells are present in an in vitro cell culture.

[0970] Item 524. The method as described in Item 523, wherein the proliferative cells are obtained from the subject.

[0971] Item 525. The method of Item 515, wherein the proliferative cell is present in the subject.

[0972] Item 526. The method of any one of items 489 to 494, wherein the neoantigen vaccine comprises at least one neoantigen mRNA.

[0973] Item 527. The method of Item 526, wherein the at least one neoantigen mRNA encodes one or more neoantigen sequences.

[0974] Item 528. The method of Item 527, wherein the neoantigen sequence is a neoantigen sequence specific to the subject.

[0975] Item 529. The method as described in Item 527, wherein the neoantigen sequence is a universal neoantigen sequence.

[0976] Item 530. The method of any one of items 527 to 529, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO: 37-65.

[0977] Item 531. The method of Item 530, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:37.

[0978] Item 532. The method of item 530, wherein the neoantigen sequence comprises the amino acid sequence of SEQ ID NO:39.

[0979] Item 533. The method of item 530, wherein the neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[0980] Item 534. The method of any one of items 527 to 529, wherein the novel antigen sequence comprises the amino acid sequence of any one of SEQ ID NO:66-93 or the antigen portion of any one of SEQ ID NO:66-93.

[0981] Item 535. The method of Item 534, wherein the novel antigen sequence comprises the amino acid sequence of SEQ ID NO:66 or the antigen portion of SEQ ID NO:66.

[0982] Item 536. The method of Item 534, wherein the novel antigen sequence comprises an amino acid sequence of any one of SEQ ID NO:74-77, or an antigenic portion of any one of SEQ ID NO:74-77.

[0983] Item 537. The method of any one of items 530 to 536, wherein the length of the neoantigen sequence and / or antigenic portion is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[0984] Item 538. The method of any one of items 530 to 537, wherein the neoantigen sequence and / or antigen portion is a personalized neoantigen vaccine for the subject.

[0985] Item 539. The method of Item 538, wherein the neoantigen sequence and / or antigenic portion has been identified by sequencing at least one neoantigen induced in the subject by administration of an effective amount of the splicing modulator, antibody-drug conjugate, or composition.

[0986] Item 540. The method of Item 538 or Item 539, wherein the neoantigen sequence and / or antigenic portion is capable of binding to at least one HLA allele expressed in the subject.

[0987] Item 541. The method of any one of items 530 to 537, wherein the neoantigen sequence and / or antigen portion is a universal neoantigen vaccine.

[0988] Item 542. The method of Item 541, wherein the neoantigen 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 subject population suffering from the neoplastic condition.

[0989] Item 543. The method of Item 541 or Item 542, wherein the neoantigen sequence and / or antigen portion is capable of eliciting a T-cell response against tumors present in at least 1%, at least 5%, or at least 10% of the subject population suffering from the neoplastic condition.

[0990] Item 544. The method of any one of items 541 to 543, wherein the at least one neoantigen mRNA encodes a neoantigen sequence induced by contacting a proliferative cell with an effective amount of the splicing regulator, antibody-drug conjugate, or composition.

[0991] Item 545. The method of any one of items 489 to 494, wherein the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable vector.

[0992] Item 546. The method of Item 545, wherein the at least one neoantigen mRNA is ligated to the pharmaceutically acceptable vector.

[0993] Item 547. The method as described in Item 545 or Item 546, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

[0994] Item 548. The method of any one of items 489 to 494, wherein the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable diluent.

[0995] Item 549. The method of any one of items 489 to 494, wherein the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable adjuvant.

[0996] Item 550. The method of any one of items 526 to 549, wherein the neoantigen mRNA is encapsulated by an encapsulating agent.

[0997] Item 551. The method of Item 550, wherein the encapsulating agent is a liposome.

[0998] Item 552. The method of Item 550, wherein the encapsulating agent is a nanoparticle.

[0999] Item 553. The method of Item 544, wherein the proliferative cells are present in an in vitro cell culture.

[1000] Item 554. The method as described in Item 553, wherein the proliferative cells are obtained from the subject.

[1001] Item 555. The method as described in Item 544, wherein the proliferative cell is present in the subject.

[1002] Item 556. The method of any one of items 451 to 469, wherein the at least one additional therapy comprises administration of a cytokine or a cytokine analogue.

[1003] Item 557. The method of Item 556, wherein the subject is intolerant, unresponsive, or poorly responsive to the cytokine or cytokine analogue when administered alone.

[1004] Item 558. The method as described in Item 556 or Item 557, wherein the cytokine or cytokine analog comprises a T-cell enhancer.

[1005] Item 559. The method of any one of items 556 to 558, wherein the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, IL-15, IFNγ and / or TNFα.

[1006] Item 560. The method of any one of items 451 to 469, wherein the at least one additional therapy comprises administering engineered tumor-targeting T cells.

[1007] Item 561. The method of any one of items 451 to 560, wherein the subject has a nonsynonymous mutation burden of about 150 mutations or less.

[1008] Item 562. The method of any one of items 451 to 561, wherein the subject has a burden of about 100 mutations or fewer nonsynonymous mutations.

[1009] Item 563. The method of any one of items 451 to 562, wherein the subject has a burden of about 50 mutations or fewer nonsynonymous mutations.

[1010] Item 564. The method of any one of items 451 to 563, wherein the proliferative condition is a hematologic malignancy or a solid tumor.

[1011] Item 565. The method as described in Item 564, wherein the hematologic malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

[1012] Item 566. The method as described in Item 564 or Item 565, wherein the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma.

[1013] Item 567. The method as described in Item 564, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary gland duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney cancer, colorectal cancer, and esophageal cancer.

[1014] Item 568. A method for identifying at least one neoantigen, the method comprising:

[1015] (a) Contacting the proliferative cells with an effective amount of a splicing regulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197;

[1016] (b) After the proliferating cells are contacted with the splicing regulator, antibody-drug conjugate, or composition, detect at least one alternatively spliced ​​mRNA transcript;

[1017] (c) Predicting the translation of at least one alternatively spliced ​​mRNA transcript into at least one peptide; and

[1018] (d) The at least one peptide is compared with 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.

[1019] Item 569. The method of Item 568, wherein detecting at least one alternatively spliced ​​mRNA transcript includes RNAseq.

[1020] Item 570. The method as described in Item 568 or Item 569, wherein predicting the translation of the at least one alternatively spliced ​​mRNA transcript includes quantifying the splice percentage change (dPSI) value of the at least one transcript.

[1021] Item 571. The method of any one of items 568 to 570, wherein predicting the translation of the at least one alternatively spliced ​​mRNA transcript comprises RiboSeq and / or ribosome mapping analysis.

[1022] Item 572. The method of any one of items 568 to 571, wherein the method further comprises evaluating the predicted major histocompatibility complex (MHC) binding of the at least one peptide.

[1023] Item 573. The method of Item 572, wherein the predicted MHC binding is determined by measuring the original affinity of the at least one peptide to predict the binding strength.

[1024] Item 574. The method as described in Item 573, wherein a primitive affinity of about 500 nM or higher predicts binding strength indicating MHC binding.

[1025] Item 575. The method of Item 572, wherein the predicted MHC binding is determined by: identifying a distribution of predicted binding strengths of a series of random peptides; and comparing the predicted binding strength of at least one peptide with the distribution.

[1026] Item 576. The method as described in Item 575, wherein the predicted binding strength in the top 2.0% of the distribution indicates weak MHC binding.

[1027] Item 577. The method as described in Item 575, wherein the predicted binding strength in the top 0.5% of the distribution indicates strong MHC binding.

[1028] Item 578. The method of any one of items 568 to 577, wherein the proliferative cells are present in an in vitro cell culture.

[1029] Item 579. The method as described in Item 578, wherein the proliferative cells are obtained from the subject.

[1030] Item 580. The method of any one of items 568 to 577, wherein the proliferative cells are present in the subject.

[1031] Item 581. The method of any one of items 568 to 580, further comprising contacting one or more additional proliferative cells to identify at least one universal neoantigen.

[1032] Item 582. A method for identifying at least one neoantigen, the method comprising:

[1033] (a) Contacting the proliferative cells with an effective amount of a splicing regulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197;

[1034] (b) After the proliferating cells have been contacted with the splicing regulator, antibody-drug conjugate, or composition, detect at least one peptide containing a possible neoantigen sequence; and

[1035] (c) The at least one peptide is compared with 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.

[1036] Item 583. The method of Item 582, wherein the method further comprises evaluating the predicted major histocompatibility complex (MHC) binding of the at least one peptide.

[1037] Item 584. The method of Item 583, wherein the predicted MHC binding is determined by measuring the original affinity of the at least one peptide to predict the binding strength.

[1038] Item 585. The method as described in Item 584, wherein a primitive affinity of about 500 nM or higher indicates MHC binding strength.

[1039] Item 586. The method of Item 583, wherein the predicted MHC binding is determined by: identifying a distribution of predicted binding strengths of a series of random peptides; and comparing the predicted binding strength of at least one peptide with the distribution.

[1040] Item 587. The method as described in Item 586, wherein the predicted binding strength in the top 2.0% of the distribution indicates weak MHC binding.

[1041] Item 588. The method as described in Item 586, wherein the predicted binding strength in the top 0.5% of the distribution indicates strong MHC binding.

[1042] Item 589. The method of any one of items 582 to 588, wherein the proliferative cells are present in an in vitro cell culture.

[1043] Item 590. The method as described in Item 589, wherein the proliferative cells are obtained from the subject.

[1044] Item 591. The method of any one of items 582 to 588, wherein the proliferative cells are present in the subject.

[1045] Item 592. The method of any one of items 582 to 591, further comprising contacting one or more additional proliferative cells to identify at least one universal neoantigen.

[1046] Item 593. A method for manufacturing a neoantigen vaccine, the method comprising:

[1047] (a) Identifying at least one neoantigen using the method described in any one of items 568 to 592; and

[1048] (b) The at least one neoantigen is formulated together with a pharmaceutically acceptable carrier, diluent or adjuvant.

[1049] Item 594. The method of Item 593, wherein the at least one neoantigen comprises the amino acid sequence of any one of SEQ ID NO:37-65.

[1050] Item 595. The method of Item 594, wherein the at least one neoantigen comprises the amino acid sequence of SEQ ID NO:37.

[1051] Item 596. The method of Item 594, wherein the at least one neoantigen comprises the amino acid sequence of SEQ ID NO:39.

[1052] Item 597. The method of Item 594, wherein the at least one neoantigen comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[1053] Item 598. The method of Item 593, wherein the at least one neoantigen comprises the amino acid sequence of any one of SEQ ID NO:66-93, or the antigenic portion of any one of SEQ ID NO:66-93.

[1054] Item 599. The method of Item 598, wherein the at least one neoantigen comprises the amino acid sequence of SEQ ID NO:66 or the antigenic portion of SEQ ID NO:66.

[1055] Item 600. The method of Item 598, wherein the at least one neoantigen comprises an amino acid sequence of any one of SEQ ID NO:74-77, or an antigenic portion of any one of SEQ ID NO:74-77.

[1056] Item 601. The method of any one of items 593 to 600, wherein the length of the at least one neoantigen and / or antigenic moiety is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[1057] Item 602. The method of any one of items 593 to 601, wherein the at least one neoantigen and / or antigen portion is linked to the pharmaceutically acceptable carrier.

[1058] Item 603. The method of any one of items 593 to 602, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines and chemokines.

[1059] Item 604. A method for treating a subject who has or is suspected of having a neoplastic condition, the method comprising:

[1060] (a) Administering to the subject an effective amount of a splicing modulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197;

[1061] (b) Detect one or more neoantigens in the subject after administration of the splicing modulator, antibody-drug conjugate, or composition;

[1062] (c) Comparing the one or more neoantigens with a set of universal neoantigens; and

[1063] (d) Administer to the subject a universal neoantigen vaccine containing at least one universal neoantigen present in the subject.

[1064] Item 605. The method of Item 604, wherein the universal neoantigen vaccine is administered alone or in combination with at least one additional therapy.

[1065] Item 606. The method of Item 605, wherein the at least one additional therapy comprises at least one, at least two, at least three, at least four, or at least five additional therapies.

[1066] Item 607. The method as described in Item 605 or Item 606, wherein the at least one additional therapy comprises repeated administration of the splice modulator, antibody-drug conjugate, or composition.

[1067] Item 608. The method of Item 607, wherein repeated administration of the splice modulator, antibody-drug conjugate, or composition is initiated prior to administration of the universal neoantigen vaccine.

[1068] Item 609. The method of Item 607, wherein the repetition of the splice modulator, antibody-drug conjugate, or composition is initiated after administration of the universal neoantigen vaccine.

[1069] Item 610. The method of Item 607, wherein repeated administration of the splice modulator, antibody-drug conjugate, or composition is initiated simultaneously with the administration of the universal neoantigen vaccine.

[1070] Item 611. The method of any one of items 607 to 610, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced compared to the amount used for initial administration.

[1071] Item 612. The method of any one of items 607 to 611, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced compared to a standard dose of the splice modulator, antibody-drug conjugate, or composition.

[1072] Item 613. The method of any one of items 607 to 612, wherein the amount of the splice modulator, antibody-drug conjugate, or composition used for repeated administration is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to a standard dose of the splice modulator, antibody-drug conjugate, or composition.

[1073] Item 614. The method of any one of items 605 to 613, wherein the at least one additional therapy includes the administration of a checkpoint inhibitor.

[1074] Item 615. The method of Item 614, wherein the administration of the checkpoint inhibitor is initiated prior to the administration of the universal neoantigen vaccine and / or repeated administration of the splice modulator, antibody-drug conjugate, or composition.

[1075] Item 616. The method of Item 614, wherein the administration of the checkpoint inhibitor is initiated after administration of the universal neoantigen vaccine and / or repetition of the splice modulator, antibody-drug conjugate, or composition.

[1076] Item 617. The method of Item 614, wherein the administration of the checkpoint inhibitor is initiated simultaneously with the administration of the universal neoantigen vaccine and / or repeated administration of the splice modulator, antibody-drug conjugate, or composition.

[1077] Item 618. The method of any one of items 614 to 617, wherein the checkpoint inhibitor is repeated at least once after the initial application.

[1078] Item 619. The method of Item 618, wherein the amount of the checkpoint inhibitor used for repeated application is reduced compared to the amount used for the initial application.

[1079] Item 620. The method of Item 618 or Item 619, wherein the amount of the checkpoint inhibitor used for repeated application is reduced compared to the standard dose of the checkpoint inhibitor.

[1080] Item 621. The method of any one of items 618 to 620, wherein the amount of the checkpoint inhibitor used for repeated application is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to a standard dose of the checkpoint inhibitor.

[1081] Item 622. The method of any one of items 614 to 621, wherein the subject is intolerant, unresponsive, or poorly responsive to the checkpoint inhibitor when administered alone.

[1082] Item 623. The method of any one of items 614 to 622, wherein the checkpoint inhibitor targets CTLA4, PD1, PDL1, OX40, CD40, GITR, LAG3, TIM3 and / or KIR.

[1083] Item 624. The method of any one of items 614 to 623, wherein the checkpoint inhibitor targets CTLA4, OX40, CD40 and / or GITR.

[1084] Item 625. The method of any one of items 614 to 624, wherein the checkpoint inhibitor comprises a cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) pathway inhibitor.

[1085] Item 626. The method as described in Item 625, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

[1086] Item 627. The method as described in Item 626, wherein the anti-CTLA4 antibody is ipilimumab.

[1087] Item 628. The method of any one of items 614 to 624, wherein the checkpoint inhibitor comprises a programmed death-1 pathway (PD1) inhibitor.

[1088] Item 629. The method of Item 628, wherein the PD1 inhibitor is an anti-PD1 antibody.

[1089] Item 630. The method as described in Item 629, wherein the anti-PD1 antibody is nivolumab.

[1090] Item 631. The method of Item 628, wherein the PD1 inhibitor is an anti-PDL1 antibody.

[1091] Item 632. The method of Item 631, wherein the anti-PDL1 antibody is atezolizumab.

[1092] Item 633. The method of any one of items 614 to 624, wherein the checkpoint inhibitor comprises a CTLA4 inhibitor and a PD1 inhibitor.

[1093] Item 634. The method as described in Item 633, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

[1094] Item 635. The method as described in Item 634, wherein the anti-CTLA4 antibody is ipilimumab.

[1095] Item 636. The method of Item 633, wherein the PD1 inhibitor is an anti-PD1 antibody.

[1096] Item 637. The method as described in Item 636, wherein the anti-PD1 antibody is nivolumab.

[1097] Item 638. The method of Item 633, wherein the PD1 inhibitor is an anti-PDL1 antibody.

[1098] Item 639. The method as described in Item 638, wherein the anti-PDL1 antibody is atezolizumab.

[1099] Item 640. The method of any one of items 604 to 639, wherein the universal neoantigen vaccine comprises at least one neoantigen peptide.

[1100] Item 641. The method of Item 640, wherein the length of the at least one neoantigen peptide is in the range of about 10 to about 50 amino acids or about 10 to about 35 amino acids.

[1101] Item 642. The method of Item 640 or Item 641, wherein the length of the at least one neoantigen peptide is in the range of about 15 to about 25 amino acids.

[1102] Item 643. The method of any one of items 640 to 642, wherein the at least one neoantigen peptide comprises one or more universal neoantigen sequences.

[1103] Item 644. The method of Item 643, wherein the universal neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:37-65.

[1104] Item 645. The method of Item 644, wherein the universal neoantigen sequence comprises the amino acid sequence of SEQ ID NO:37.

[1105] Item 646. The method of Item 644, wherein the universal neoantigen sequence comprises the amino acid sequence of SEQ ID NO:39.

[1106] Item 647. The method of Item 644, wherein the universal neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[1107] Item 648. The method of Item 643, wherein the universal neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:66-93, or the antigen portion of any one of SEQ ID NO:66-93.

[1108] Item 649. The method of Item 648, wherein the universal neoantigen sequence comprises the amino acid sequence of SEQ ID NO:66 or the antigenic portion of SEQ ID NO:66.

[1109] Item 650. The method of Item 648, wherein the universal neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:74-77, or the antigen portion of any one of SEQ ID NO:74-77.

[1110] Item 651. The method of any one of items 643 to 650, wherein the length of the universal neoantigen sequence and / or antigenic portion is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[1111] Item 652. The method of any one of items 643 to 651, wherein the universal neoantigen 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 subject population suffering from the neoplastic condition.

[1112] Item 653. The method of any one of items 643 to 652, wherein the universal neoantigen sequence and / or antigen 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 subject population suffering from the neoplastic condition.

[1113] Item 654. The method of any one of items 604 to 639, wherein the universal neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable carrier.

[1114] Item 655. The method of Item 654, wherein the at least one neoantigen peptide is linked to the pharmaceutically acceptable carrier.

[1115] Item 656. The method as described in Item 654 or Item 655, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

[1116] Item 657. The method of any one of items 654 to 656, wherein the neoantigen peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

[1117] Item 658. The method of any one of items 654 to 656, wherein the neoantigen peptide and the pharmaceutically acceptable vector are expressed as a fusion protein.

[1118] Item 659. The method of any one of items 604 to 639, wherein the universal neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable diluent.

[1119] Item 660. The method of any one of items 604 to 639, wherein the universal neoantigen vaccine comprises at least one neoantigen peptide and a pharmaceutically acceptable adjuvant.

[1120] Item 661. The method of any one of items 604 to 639, wherein the universal neoantigen vaccine comprises at least one neoantigen mRNA.

[1121] Item 662. The method of Item 661, wherein the at least one neoantigen mRNA encodes one or more universal neoantigen sequences.

[1122] Item 663. The method of Item 662, wherein the universal neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:37-65.

[1123] Item 664. The method of Item 663, wherein the universal neoantigen sequence comprises the amino acid sequence of SEQ ID NO:37.

[1124] Item 665. The method of Item 663, wherein the universal neoantigen sequence comprises the amino acid sequence of SEQ ID NO:39.

[1125] Item 666. The method of Item 663, wherein the universal neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[1126] Item 667. The method of Item 662, wherein the universal neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:66-93, or the antigen portion of any one of SEQ ID NO:66-93.

[1127] Item 668. The method of Item 667, wherein the universal neoantigen sequence comprises the amino acid sequence of SEQ ID NO:66 or the antigen portion of SEQ ID NO:66.

[1128] Item 669. The method of Item 667, wherein the universal neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:74-77, or the antigen portion of any one of SEQ ID NO:74-77.

[1129] Item 670. The method of any one of items 662 to 669, wherein the length of the universal neoantigen sequence and / or antigenic portion is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[1130] Item 671. The method of any one of items 662 to 670, wherein the universal neoantigen 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 subject population suffering from the neoplastic condition.

[1131] Item 672. The method of any one of items 662 to 671, wherein the universal neoantigen sequence and / or antigen 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 subject population suffering from the neoplastic condition.

[1132] Item 673. The method of any one of items 604 to 639, wherein the universal neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable vector.

[1133] Item 674. The method of Item 673, wherein the at least one neoantigen mRNA is ligated to the pharmaceutically acceptable vector.

[1134] Item 675. The method as described in Item 673 or Item 674, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

[1135] Item 676. The method of any one of items 604 to 639, wherein the universal neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable diluent.

[1136] Item 677. The method of any one of items 604 to 639, wherein the universal neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable adjuvant.

[1137] Item 678. The method of any one of items 661 to 677, wherein the neoantigen mRNA is encapsulated by an encapsulating agent.

[1138] Item 679. The method as described in Item 678, wherein the encapsulating agent is a liposome.

[1139] Item 680. The method of Item 678, wherein the encapsulating agent is a nanoparticle.

[1140] Item 681. The method of any one of items 604 to 680, wherein the subject has a nonsynonymous mutation burden of about 150 mutations or less.

[1141] Item 682. The method of any one of items 604 to 681, wherein the subject has a burden of about 100 mutations or fewer nonsynonymous mutations.

[1142] Item 683. The method of any one of items 604 to 682, wherein the subject has a nonsynonymous mutation burden of about 50 mutations or less.

[1143] Item 684. The method of any one of items 604 to 683, wherein the proliferative condition is a hematologic malignancy or a solid tumor.

[1144] Item 685. The method as described in Item 684, wherein the hematologic malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma.

[1145] Item 686. The method as described in Item 684 or Item 685, wherein the hematologic malignancy is selected from acute myeloid leukemia and multiple myeloma.

[1146] Item 687. The method as described in Item 684, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct carcinoma, melanoma, colon cancer, cervical cancer, pancreatic cancer, kidney cancer, colorectal cancer, and esophageal cancer.

[1147] Item 688. The method of any one of items 303 to 687, wherein the antibody-drug conjugate comprises an antibody-drug conjugate having formula (I):

[1148] Ab-(LD) p (I)

[1149] in

[1150] Ab is an antibody or antigen-binding fragment that targets neoplastic cells;

[1151] D is D2, D1, D4, D12, or D15;

[1152] L is the connector that covalently links Ab to D; and

[1153] p is an integer from 1 to 15.

[1154] Item 689. The method of Item 688, wherein the antibody or antigen-binding fragment targets cells expressing HER2.

[1155] Item 690. The method of Item 689, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

[1156] Item 691. The method of Item 689 or Item 690, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[1157] Item 692. The method of any one of items 689 to 691, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[1158] Item 693. The method of Item 688, wherein the antibody or antigen-binding fragment targets cells expressing CD138.

[1159] Item 694. The method of Item 693, wherein the antibody or antigen-binding fragment is an anti-CD138 antibody or antigen-binding fragment.

[1160] Item 695. The method of Item 693 or Item 694, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:10 (LCDR1), SEQ ID NO:11 (LCDR2), and SEQ ID NO:12 (LCDR3).

[1161] Item 696. The method of any one of items 693 to 695, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:21 and a light chain variable region containing the amino acid sequence of SEQ ID NO:22.

[1162] Item 697. The method as described in Item 688, wherein the antibody or antigen-binding fragment targets cells expressing EPHA2.

[1163] Item 698. The method of Item 697, wherein the antibody or antigen-binding fragment is an anti-EPHA2 antibody or antigen-binding fragment.

[1164] Item 699. The method of Item 697 or Item 698, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:16 (LCDR1), SEQ ID NO:17 (LCDR2), and SEQ ID NO:18 (LCDR3).

[1165] Item 700. The method of any one of items 697 to 699, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:23 and a light chain variable region containing the amino acid sequence of SEQ ID NO:24.

[1166] Item 701. The method of any one of items 688 to 700, wherein the joint is a pyrolytic joint.

[1167] Item 702. The method of Item 701, wherein the pyrolytic connector comprises MC-Val-Cit-pABC.

[1168] Item 703. The method of Item 701, wherein the pyrolytic connector comprises MC-Val-Ala-pABC.

[1169] Item 704. The method of Item 701, wherein the pyrolytic connector comprises MC-Val-Ala-pAB.

[1170] Item 705. The method of any one of items 688 to 700, wherein the joint is a non-split joint.

[1171] Item 706. The method of Item 705, wherein the non-disintegrable connector comprises Mal-Hex.

[1172] Item 707. The method as described in Item 705, wherein the non-disintegrable connector comprises Mal-Et.

[1173] Item 708. The method of Item 705, wherein the non-disintegrable connector comprises Mal-Et-O-Et.

[1174] Item 709. The method of any one of items 688 to 708, wherein the splice modifier comprises D2:

[1175]

[1176] Item 710. The method of Item 709, wherein the splice modifier comprises D2 and the joint comprises MC-Val-Ala-pAB.

[1177] Item 711. The method of any one of items 688 to 708, wherein the splice modifier comprises D1:

[1178]

[1179] Item 712. The method of Item 711, wherein the splice modifier comprises D1 and the joint comprises MC-Val-Cit-pABC.

[1180] Item 713. The method of any one of items 688 to 708, wherein the splice modifier comprises a splice modifier having formula (II):

[1181] or a pharmaceutically acceptable salt thereof, wherein:

[1182] R 1Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[1183] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[1184] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[1185] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups and -NR 15 R 16 ;

[1186] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and -C(=O)-OR 17 ;

[1187] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[1188] Z is selected from

[1189] where R 1 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups.

[1190] where R 6 and R 7 At least one of them is hydrogen.

[1191] Item 714. The method of any one of items 688 to 708, wherein the splice modifier comprises a splice modifier having formula (IV):

[1192] or a pharmaceutically acceptable salt thereof, wherein:

[1193] R 1 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, -OC(=O)-(C1-C6 alkyl) groups, and -CD3;

[1194] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, and -OC(=O)-(C1-C6 alkyl) groups; and

[1195] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[1196] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups and -NR 15 R 16 ;

[1197] R 15 and R 16 Each is independently selected from hydrogen, R 17 、-C(=O)-R 17 and

[1198] -C(=O)-OR 17 ;and

[1199] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[1200] where R 1 R 3 R 4 R 5 R 6 R 7 R 8 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups.

[1201] where R 6 and R 7 At least one of them is hydrogen.

[1202] Item 715. The method of any one of items 688 to 708, wherein the splice modifier comprises a splice modifier having formula (VI):

[1203] or a pharmaceutically acceptable salt thereof, wherein:

[1204] R 1 and R 9 Each group is independently selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups, -OC(=O)-(C1-C6 alkyl) groups, and -CD3;

[1205] R 3Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[1206] R 4 R 5 and R 8 Each group is independently selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups;

[1207] R 6 and R 7 Each is independently selected from hydrogen, -OR 17 -OC(=O)-R 17 -OC(=O)-NR 15 R 16 C1-C6 alkyl groups, -NR 15 R 16 and connectors;

[1208] R 10 Selected from hydrogen, C1-C6 alkyl groups, -C(=O)-(C1-C6 alkyl) groups, and -CD3;

[1209] R 15 and R 16 Each is independently selected from hydrogen, R 17 -C(=O)-R 17 and -C(=O)-OR 17 ;

[1210] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups; and

[1211] a is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[1212] Where R 1 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 15 R 16 and R 17Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups;

[1213] where R 6 and R 7 At least one of them is hydrogen; and

[1214] where R 1 and R 9 They cannot both be absent at the same time.

[1215] Item 716. The method of any one of items 688 to 708, wherein the splice modifier comprises a splice modifier having formula (VIII):

[1216] or a pharmaceutically acceptable salt thereof, wherein:

[1217] R 1 Selected from non-existent, hydrogen, C1-C6 alkyl group, C1-C6 alkylalkoxy group, C1-C6 alkylamino group, C1-C6 alkylcarboxylic acid group, C1-C6 alkylhydroxy group, C3-C8 cycloalkyl group, benzyl group, C3-C8 heterocyclic group, -OC(=O)-(C1-C6 alkyl) group and -CD3;

[1218] R 3 Selected from hydrogen, C1-C6 alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylamino groups, C1-C6 alkylcarboxylic acid groups, C1-C6 alkylhydroxy groups, C3-C8 cycloalkyl groups, benzyl groups, C3-C8 heterocyclic groups and -OC(=O)-(C1-C6 alkyl) groups;

[1219] R 4 Selected from hydrogen, hydroxyl groups, -O-(C1-C6 alkyl) groups, -OC(=O)-(C1-C6 alkyl) groups, and C1-C6 alkyl groups; and

[1220] R 10 Selected from 3- to 10-membered carbon rings and 3- to 10-membered heterocycles, each of which is composed of 0 to 3 R... a Replace, where each R a Independently selected from halogens, C1-C6 alkyl groups, -O-(C1-C6)alkyl groups, C1-C6 alkylalkoxy groups, C1-C6 alkylhydroxy groups, and -S (=O). w-(4- to 7-membered heterocycles), 4- to 7-membered carbon rings and 4- to 7-membered heterocycles;

[1221] R 15 and R 16 Each is independently selected from hydrogen, R 17 -C(=O)-R 17 and -C(=O)-OR 17 ;and

[1222] R 17 Selected from hydrogen, C1-C6 alkyl groups, C3-C8 cycloalkyl groups, benzyl groups, and C3-C8 heterocyclic groups;

[1223] Where R 1 R 3 R 4 R 10 R 15 R 16 and R 17 Each is independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, C1-C6 alkyl group, -O-(C1-C6 alkyl) group, -NR 15 R 16 C3-C8 cycloalkyl groups, C1-C6 alkylhydroxy groups, C1-C6 alkylalkoxy groups, benzyl groups, and C3-C8 heterocyclic groups; and

[1224] Each R a Independently substituted by 0 to 3 groups independently selected from the following: halogen, hydroxyl group, -NR 15 R 16 C1-C6 alkyl groups, -(C=O)-(C1-C6 alkyl) groups, -(C=O)-(C1-C6 alkyl)-(C3-C 10 Heterocyclic groups, -S (=O) w -(C3-C8 heterocyclic group) and C1-C6 alkylcarboxylic acid group, each of which is substituted by 0, 1 or 2 independently selected from the following groups: halogen, hydroxyl group, -NR 15 R 16 and C1-C3 alkyl groups; and

[1225] w is 0, 1, or 2.

[1226] Item 717. The method of any one of items 688 to 708, wherein the splice modifier comprises a modifier of the SF3b complex.

[1227] Item 718. The method of Item 717, wherein the splice modulator comprises prasadiene lactone or a prasadiene lactone derivative.

[1228] Item 719. The method of Item 717 or Item 718, wherein the splicing modifier comprises prasadiene lactone D or a prasadiene lactone D derivative.

[1229] Item 720. The method of Item 719, wherein the prasadiene lactone D or its derivative comprises D2, D1, D4, D8, D10, D11 (E7107), D20, D21, D22, D12 or D25.

[1230] Item 721. The method as described in Item 719 or Item 720, wherein the prasadiene lactone D or its derivative comprises D2.

[1231] Item 722. The method as described in Item 719 or Item 720, wherein the prasadiene lactone D or its derivative comprises D1.

[1232] Item 723. The method as described in Item 719 or Item 720, wherein the prasadiene lactone D or its derivative is a zwitterionic prasadiene lactone D or its derivative.

[1233] Item 724. The method of Item 723, wherein the zwitterionic prasadiene lactone D or its derivative comprises D22 or D25.

[1234] Item 725. The method of Item 717 or Item 718, wherein the splicing modifier comprises prasadiene lactone B or a prasadiene lactone B derivative.

[1235] Item 726. The method of Item 725, wherein the prasadiene lactone B or its derivative comprises D9, D18, D19 or D13.

[1236] Item 727. The method as described in Item 717 or Item 718, wherein the splice modifier comprises arylpradolide lactone.

[1237] Item 728. The method of Item 727, wherein the aryl prasadiene lactone comprises D15, D14, D16, D17, D26 or D33.

[1238] Item 729. The method as described in Item 727 or Item 728, wherein the arylpradolide is an amphoteric arylpradolide.

[1239] Item 730. The method of Item 729, wherein the zwitterionic arylpradolide comprises D33.

[1240] Item 731. The method of any one of items 688 to 708, wherein the splice modifier comprises D4, D12, D15, D8, D9, D10, D13, D18, D19, D20, D21, D22, D25 or D33.

[1241] Item 732. The method as described in any one of items 688 to 731, wherein p is 2 to 8.

[1242] Item 733. The method as described in any one of items 688 to 732, wherein p is 4 to 8.

[1243] Item 734. The method as described in any one of items 688 to 733, wherein p is 4 or 8.

[1244] Item 735. A neoantigen vaccine comprising at least one neoantigen peptide, wherein the at least one neoantigen peptide comprises a modified or novel neoantigen sequence induced by contacting a proliferative cell with an effective amount of a splicing regulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197.

[1245] Item 736. The neoantigen vaccine as described in Item 735, wherein the length of the at least one neoantigen peptide is in the range of about 10 to about 50 amino acids or about 10 to about 35 amino acids.

[1246] Item 737. The neoantigen vaccine as described in Item 735 or Item 736, wherein the length of the at least one neoantigen peptide is in the range of about 15 to about 25 amino acids.

[1247] Item 738. The neoantigen vaccine as described in any one of items 735 to 737, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO: 37-65.

[1248] Item 739. The neoantigen vaccine as described in Item 738, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:37.

[1249] Item 740. The neoantigen vaccine as described in Item 738, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:39.

[1250] Item 741. The neoantigen vaccine as described in Item 738, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[1251] Item 742. The neoantigen vaccine as described in any one of items 735 to 737, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO: 66-93, or the antigen portion of any one of SEQ ID NO: 66-93.

[1252] Item 743. The neoantigen vaccine as described in item 742, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:66 or the antigen portion of SEQ ID NO:66.

[1253] Item 744. The neoantigen vaccine as described in Item 742, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:74-77, or the antigen portion of any one of SEQ ID NO:74-77.

[1254] Item 745. The neoantigen vaccine as described in any one of items 735 to 744, wherein the length of the modified or novel neoantigen sequence and / or antigenic portion is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[1255] Item 746. The neoantigen vaccine as described in any one of items 735 to 745, wherein the neoantigen vaccine further comprises a pharmaceutically acceptable carrier.

[1256] Item 747. The neoantigen vaccine as described in Item 746, wherein the at least one neoantigen peptide is linked to the pharmaceutically acceptable carrier.

[1257] Item 748. The neoantigen vaccine as described in Item 746 or Item 747, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

[1258] Item 749. The neoantigen vaccine as described in any one of items 746 to 748, wherein the neoantigen peptide and the pharmaceutically acceptable carrier are covalently linked via a connector.

[1259] Item 750. The neoantigen vaccine as described in any one of items 746 to 748, wherein the neoantigen peptide and the pharmaceutically acceptable vector are expressed as a fusion protein.

[1260] Item 751. The neoantigen vaccine as described in any one of items 735 to 745, wherein the neoantigen vaccine further comprises a pharmaceutically acceptable diluent.

[1261] Item 752. The neoantigen vaccine as described in any one of items 735 to 745, wherein the neoantigen vaccine further comprises a pharmaceutically acceptable adjuvant.

[1262] Item 753. The neoantigen vaccine as described in any one of items 735 to 752, wherein the proliferative cells are present in an in vitro cell culture.

[1263] Item 754. The neoantigen vaccine as described in Item 753, wherein the proliferative cells are obtained from the subject.

[1264] Item 755. The neoantigen vaccine as described in any one of items 735 to 752, wherein the proliferative cells are present in the subject.

[1265] Item 756. A neoantigen vaccine comprising at least one neoantigen mRNA, wherein the at least one neoantigen mRNA encodes a modified or novel neoantigen sequence induced by contacting a proliferative cell with an effective amount of a splicing regulator, an antibody-drug conjugate as described in any one of items 1 to 193, or a composition as described in any one of items 194 to 197.

[1266] Item 757. The neoantigen vaccine as described in Item 756, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:37-65.

[1267] Item 758. The neoantigen vaccine as described in Item 757, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:37.

[1268] Item 759. The neoantigen vaccine as described in Item 757, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:39.

[1269] Item 760. The neoantigen vaccine as described in Item 757, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:46-49.

[1270] Item 761. The neoantigen vaccine as described in Item 756, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:66-93, or the antigen portion of any one of SEQ ID NO:66-93.

[1271] Item 762. The neoantigen vaccine as described in Item 761, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of SEQ ID NO:66 or the antigen portion of SEQ ID NO:66.

[1272] Item 763. The neoantigen vaccine as described in Item 761, wherein the modified or novel neoantigen sequence comprises the amino acid sequence of any one of SEQ ID NO:74-77, or the antigen portion of any one of SEQ ID NO:74-77.

[1273] Item 764. The neoantigen vaccine as described in any one of items 756 to 763, wherein the length of the modified or novel neoantigen sequence and / or antigenic moiety is in the range of about 10 to about 50, about 10 to about 35, about 15 to about 25, or about 10 to about 20 amino acids.

[1274] Item 765. The neoantigen vaccine as described in any one of items 756 to 764, wherein the neoantigen vaccine further comprises a pharmaceutically acceptable carrier.

[1275] Item 766. The neoantigen vaccine as described in Item 765, wherein the at least one neoantigen mRNA is ligated to the pharmaceutically acceptable vector.

[1276] Item 767. The neoantigen vaccine as described in Item 765 or Item 766, wherein the pharmaceutically acceptable carrier is selected from peptides, serum albumin, keyhole hemocyanin, immunoglobulins, thyroglobulin, ovalbumin, toxoids or attenuated toxoid derivatives, cytokines, and chemokines.

[1277] Item 768. The neoantigen vaccine as described in any one of items 756 to 764, wherein the neoantigen vaccine further comprises a pharmaceutically acceptable diluent.

[1278] Item 769. The neoantigen vaccine as described in any one of items 756 to 764, wherein the neoantigen vaccine further comprises a pharmaceutically acceptable adjuvant.

[1279] Item 770. The neoantigen vaccine as described in any one of items 756 to 769, wherein the neoantigen mRNA is encapsulated by an encapsulating agent.

[1280] Item 771. The neoantigen vaccine as described in Item 770, wherein the encapsulating agent is a liposome.

[1281] Item 772. The neoantigen vaccine as described in Item 770, wherein the encapsulating agent is nanoparticles.

[1282] Item 773. The neoantigen vaccine as described in any one of items 756 to 772, wherein the proliferative cells are present in an in vitro cell culture.

[1283] Item 774. The neoantigen vaccine as described in Item 773, wherein the proliferative cells are obtained from the subject.

[1284] Item 775. The neoantigen vaccine as described in any one of items 756 to 772, wherein the proliferative cells are present in the subject.

[1285] Item 776. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing MSLN.

[1286] Item 777. The antibody-drug conjugate as described in Item 776, wherein the antibody or antigen-binding fragment is an anti-MSLN antibody or antigen-binding fragment.

[1287] Item 778. An antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing FOLH1.

[1288] Item 779. The antibody-drug conjugate as described in Item 778, wherein the antibody or antigen-binding fragment is an anti-FOLH1 antibody or antigen-binding fragment.

[1289] Item 780. An antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing CDH6.

[1290] Item 781. The antibody-drug conjugate as described in Item 780, wherein the antibody or antigen-binding fragment is an anti-CDH6 antibody or antigen-binding fragment.

[1291] Item 782. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing CEACAM5.

[1292] Item 783. The antibody-drug conjugate as described in Item 782, wherein the antibody or antigen-binding fragment is an anti-CEACAM5 antibody or antigen-binding fragment.

[1293] Item 784. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing CFC1B.

[1294] Item 785. The antibody-drug conjugate as described in Item 784, wherein the antibody or antigen-binding fragment is an anti-CFC1B antibody or antigen-binding fragment.

[1295] Item 786. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing ENPP3.

[1296] Item 787. The antibody-drug conjugate as described in Item 786, wherein the antibody or antigen-binding fragment is an anti-ENPP3 antibody or antigen-binding fragment.

[1297] Item 788. An antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing FOLR1.

[1298] Item 789. The antibody-drug conjugate as described in Item 788, wherein the antibody or antigen-binding fragment is an anti-FOLR1 antibody or antigen-binding fragment.

[1299] Item 790. An antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing HAVCR1.

[1300] Item 791. The antibody-drug conjugate as described in Item 790, wherein the antibody or antigen-binding fragment is an anti-HAVCR1 antibody or antigen-binding fragment.

[1301] Item 792. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing KIT.

[1302] Item 793. The antibody-drug conjugate as described in Item 792, wherein the antibody or antigen-binding fragment is an anti-KIT antibody or antigen-binding fragment.

[1303] Item 794. An antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing MET.

[1304] Item 795. The antibody-drug conjugate as described in Item 794, wherein the antibody or antigen-binding fragment is an anti-MET antibody or antigen-binding fragment.

[1305] Item 796. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing MUC16.

[1306] Item 797. The antibody-drug conjugate as described in Item 796, wherein the antibody or antigen-binding fragment is an anti-MUC16 antibody or antigen-binding fragment.

[1307] Item 798. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing SLC39A6.

[1308] Item 799. The antibody-drug conjugate as described in Item 798, wherein the antibody or antigen-binding fragment is an anti-SLC39A6 antibody or antigen-binding fragment.

[1309] Item 800. An antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing SLC44A4.

[1310] Item 801. The antibody-drug conjugate as described in Item 800, wherein the antibody or antigen-binding fragment is an anti-SLC44A4 antibody or antigen-binding fragment.

[1311] Item 802. The antibody-drug conjugate as described in any one of items 1 to 7, wherein the antibody or antigen-binding fragment targets cells expressing STEAP1.

[1312] Item 803. The antibody-drug conjugate as described in Item 802, wherein the antibody or antigen-binding fragment is an anti-STEAP1 antibody or antigen-binding fragment.

[1313] Item 804. The antibody-drug conjugate as described in Item 29, wherein the amino acid unit comprises glutamine-valine-citrulline (Glu-Val-Cit).

[1314] Item 805. The antibody-drug conjugate as described in Item 29, wherein the amino acid unit comprises alanine-alanine-asparagine (Ala-Ala-Asn).

[1315] Item 806. The antibody-drug conjugate as described in Item 48 or 49, wherein the cleavable peptide moiety or amino acid unit comprises Glu-Val-Cit.

[1316] Item 807. The antibody-drug conjugate as described in Item 48 or 49, wherein the cleavable peptide moiety or amino acid unit comprises Ala-Ala-Asn.

[1317] Item 808. The antibody-drug conjugate as described in any one of items 55 to 58, wherein the linker comprises MC-Glu-Val-Cit.

[1318] Item 809. The antibody-drug conjugate as described in any one of items 55 to 58, wherein the linker comprises MC-Ala-Ala-Asn.

[1319] Item 810. The antibody-drug conjugate as described in any one of items 67 to 71, wherein the linker comprises Glu-Val-Cit-pABC.

[1320] Item 811. The antibody-drug conjugate as described in any one of items 67 to 71, wherein the linker comprises Ala-Ala-Asn-pABC.

[1321] Item 812. The antibody-drug conjugate as described in any one of items 74 to 78, wherein the linker comprises Glu-Val-Cit-pAB.

[1322] Item 813. The antibody-drug conjugate as described in any one of items 74 to 78, wherein the linker comprises Ala-Ala-Asn-pAB.

[1323] Item 814. The antibody-drug conjugate as described in Item 117 or Item 118, wherein the prasadiene lactone D or its derivative comprises D4.

[1324] Item 815. The antibody-drug conjugate as described in Item 117 or Item 118, wherein the prasadiene lactone D or its derivative comprises D12.

[1325] Item 816. The antibody-drug conjugate as described in Item 125 or Item 126, wherein the arylpradoside lactone comprises D15.

[1326] Item 817. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator comprises D4:

[1327]

[1328] Item 818. The antibody-drug conjugate as described in Item 817, wherein the splicing modulator comprises D4 and the linker comprises MC-Val-Cit-pABC.

[1329] Item 819. The antibody-drug conjugate as described in Item 817, wherein the splicing modulator comprises D4 and the linker comprises MC-β-glucuronide.

[1330] Item 820. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator comprises D12:

[1331]

[1332] Item 821. The antibody-drug conjugate as described in 820, wherein the splicing modulator comprises D12 and the linker comprises MC-Val-Cit-pABC.

[1333] Item 822. The antibody-drug conjugate as described in Item 820, wherein the splicing modulator comprises D12 and the linker comprises MC-β-glucuronide.

[1334] Item 823. The antibody-drug conjugate as described in any one of items 1 to 106, wherein the splicing modulator comprises D15:

[1335]

[1336] Item 824. The antibody-drug conjugate as described in 823, wherein the splicing modulator comprises D15 and the linker comprises MC-Val-Ala-pAB.

[1337] Item 825. The antibody-drug conjugate as described in Item 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.

[1338] Item 826. The antibody-drug conjugate as described in Item 156, 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.

[1339] Item 827. An antibody-drug conjugate having formula (I),

[1340] Ab-(LD) p (I)

[1341] in

[1342] Ab is an antibody or antigen-binding fragment that targets neoplastic cells;

[1343] D is D4;

[1344] L is the connector that covalently links Ab to D; and

[1345] p is an integer from 1 to 15.

[1346] Item 828. The antibody-drug conjugate as described in Item 827, wherein the antibody or antigen-binding fragment targets cells expressing HER2.

[1347] Item 829. The antibody-drug conjugate as described in Item 828, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

[1348] Item 830. The antibody-drug conjugate as described in Item 828 or Item 829, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[1349] Item 831. The antibody-drug conjugate as described in any one of items 828 to 830, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[1350] Item 832. The antibody-drug conjugate as described in any one of items 828 to 831, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[1351] Item 833. The antibody-drug conjugate as described in any one of items 828 to 832, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[1352] Item 834. The antibody-drug conjugate as described in Item 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.

[1353] Item 835. An antibody-drug conjugate having formula (I),

[1354] Ab-(LD) p (I)

[1355] in

[1356] Ab is an antibody or antigen-binding fragment that targets neoplastic cells;

[1357] D is D12;

[1358] L is the connector that covalently links Ab to D; and

[1359] p is an integer from 1 to 15.

[1360] Item 836. The antibody-drug conjugate as described in Item 835, wherein the antibody or antigen-binding fragment targets cells expressing HER2.

[1361] Item 837. The antibody-drug conjugate as described in Item 836, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

[1362] Item 838. The antibody-drug conjugate as described in Item 836 or Item 837, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[1363] Item 839. The antibody-drug conjugate as described in any one of items 836 to 838, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[1364] Item 840. The antibody-drug conjugate as described in any one of items 836 to 839, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[1365] Item 841. The antibody-drug conjugate as described in any one of items 836 to 840, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[1366] Item 842. The antibody-drug conjugate as described in Item 835, 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.

[1367] Item 843. An antibody-drug conjugate having formula (I),

[1368] Ab-(LD) p (I)

[1369] in

[1370] Ab is an antibody or antigen-binding fragment that targets neoplastic cells;

[1371] D is D15;

[1372] L is the connector that covalently links Ab to D; and

[1373] p is an integer from 1 to 15.

[1374] Item 844. The antibody-drug conjugate as described in Item 843, wherein the antibody or antigen-binding fragment targets cells expressing HER2.

[1375] Item 845. The antibody-drug conjugate as described in Item 844, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

[1376] Item 846. The antibody-drug conjugate as described in Item 844 or Item 845, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[1377] Item 847. The antibody-drug conjugate as described in any one of items 844 to 846, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[1378] Item 848. The antibody-drug conjugate as described in any one of items 844 to 847, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[1379] Item 849. The antibody-drug conjugate as described in any one of items 844 to 848, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain.

[1380] Item 850. The antibody-drug conjugate as described in Item 843, 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.

[1381] Item 851. The antibody-drug conjugate as described in any one of items 827 to 850, wherein the linker is a cleavable linker.

[1382] Item 852. The antibody-drug conjugate as described in Item 851, wherein the cleavable linker comprises MC-Val-Cit-pABC.

[1383] Item 853. The antibody-drug conjugate as described in Item 851, wherein the cleavable linker comprises MC-Val-Ala-pABC.

[1384] Item 854. The antibody-drug conjugate as described in Item 851, wherein the cleavable linker comprises MC-Val-Ala-pAB.

[1385] Item 855. The antibody-drug conjugate as described in Item 851, wherein the cleavable linker comprises MC-Glu-Val-Cit-pABC.

[1386] Item 856. The antibody-drug conjugate as described in Item 851, wherein the cleavable linker comprises MC-Ala-Ala-Asn-pABC.

[1387] Item 857. The antibody-drug conjugate as described in Item 851, wherein the cleavable linker comprises MC-β-glucuronide.

[1388] Item 858. The antibody-drug conjugate as described in any one of items 827 to 850, wherein the linker is a non-cleavable linker.

[1389] Item 859. The antibody-drug conjugate as described in Item 858, wherein the cleavable linker comprises Mal-Hex.

[1390] Item 860. The antibody-drug conjugate as described in Item 858, wherein the cleavable linker comprises Mal-Et.

[1391] Item 861. The antibody-drug conjugate as described in Item 858, wherein the non-cleavable linker comprises Mal-Et-O-Et.

[1392] Item 862. The antibody-drug conjugate as described in any one of items 827 to 861, wherein p is 1 to 10.

[1393] Item 863. The antibody-drug conjugate as described in any one of items 827 to 862, wherein p is 2 to 8.

[1394] Item 864. The antibody-drug conjugate as described in any one of items 827 to 863, wherein p is 4 to 8.

[1395] Item 865. An antibody-drug conjugate as described in any one of items 827 to 864, wherein p is 4.

[1396] Item 866. An antibody-drug conjugate as described in any one of items 827 to 864, wherein p is 8.

[1397] Item 867. The antibody-drug conjugate as described in Item 181, wherein the cleavable linker comprises MC-Glu-Val-Cit-pABC.

[1398] Item 868. The antibody-drug conjugate as described in Item 181, wherein the cleavable linker comprises MC-Ala-Ala-Asn-pABC.

[1399] Item 869. A composition comprising a plurality of copies of an antibody-drug conjugate as described in any one of items 776 to 868, wherein the average p of the antibody-drug conjugate in the composition is from about 3.5 to about 5.5.

[1400] Item 870. A composition comprising a plurality of copies of an antibody-drug conjugate as described in any one of items 776 to 868, wherein the average p of the antibody-drug conjugate in the composition is about 7 to about 9.

[1401] Item 871. A method of treating a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate as described in any one of items 776 to 868 or a composition as described in item 869 or 870.

[1402] Item 872. A method for reducing or inhibiting tumor growth in a subject who has or is suspected of having a neoplastic condition, the method comprising administering to the subject a therapeutically effective amount of an antibody-drug conjugate as described in any one of items 776 to 868 or a composition as described in item 869 or 870.

[1403] Item 873. A method for determining whether a subject with or suspected of having a neoplastic condition will respond to treatment with an antibody-drug conjugate as described in any one of items 776 to 868 or a composition as described in item 869 or 870, the method comprising providing a biological sample from the subject and contacting the biological sample with an antibody-drug conjugate as described in any one of items 776 to 868 or a composition as described in item 869 or 870.

[1404] Item 874. The antibody-drug conjugate as described in any one of items 776 to 868 or the composition as described in item 869 or 870, for the treatment of adenoid diseases.

[1405] Item 875. Use of the antibody-drug conjugate as described in any one of items 776 to 868 or the composition as described in item 869 or 870 in the treatment of aplastic conditions.

[1406] Item 876. Use of the antibody-drug conjugate as described in any one of items 776 to 868 or the composition as described in item 869 or 870 in a method of manufacturing a medicament for treating aplastic conditions.

[1407] Item 877. A pharmaceutical composition comprising an antibody-drug conjugate as described in any one of items 776 to 868 or a composition as described in item 869 or 870, and a pharmaceutically acceptable carrier.

[1408] Item 878. A method for producing an antibody-drug conjugate as described in any one of items 776 to 868, the method comprising reacting an antibody or antigen-binding fragment with a linker conjugated to a splice modulator under conditions that allow conjugation.

[1409] Item 879. The method of any one of items 303 to 687, wherein the antibody-drug conjugate comprises an antibody-drug conjugate of any one of items 776 to 868.

[1410] Item 880. The method of Item 688, 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.

[1411] Item 881. The method of Item 701, wherein the pyrolytic connector comprises MC-Glu-Val-Cit-pABC.

[1412] Item 882. The method of Item 701, wherein the pyrolytic connector comprises MC-Ala-Ala-Asn-pABC.

[1413] Item 883. The method of Item 701, wherein the cleavable linker comprises MC-β-glucuronide.

[1414] Item 884. The method of any one of items 688 to 708, wherein the splice modifier comprises D4:

[1415]

[1416] Item 885. The method of Item 884, wherein the splice modifier comprises D4 and the joint comprises MC-Val-Cit-pABC.

[1417] Item 886. The method of Item 884, wherein the splice modulator comprises D4 and the splice comprises MC-β-glucuronide.

[1418] Item 887. The method of any one of items 688 to 708, wherein the splice modifier comprises D12:

[1419]

[1420] Item 888. The method of Item 887, wherein the splice modifier comprises D12 and the joint comprises MC-Val-Cit-pABC.

[1421] Item 889. The method of Item 887, wherein the splice modulator comprises D12 and the splice comprises MC-β-glucuronide.

[1422] Item 890. The method of any one of items 688 to 708, wherein the splice modifier comprises D15:

[1423]

[1424] Item 891. The method of Item 890, wherein the splice modifier comprises D15 and the connector comprises MC-Val-Ala-pAB.

[1425] Item 892. The method as described in Item 719 or Item 720, wherein the prasadiene lactone D or its derivative comprises D4.

[1426] Item 893. The method as described in Item 719 or Item 720, wherein the prasadiene lactone D or its derivative comprises D12.

[1427] Item 894. The method as described in Item 727 or Item 728, wherein the aryl prasadiene lactone comprises D15.

[1428] Item 895. A neoantigen vaccine comprising at least one neoantigen peptide, wherein the at least one neoantigen peptide comprises a modified or novel neoantigen sequence induced by contacting a proliferative cell with an effective amount of a splicing regulator, an antibody-drug conjugate as described in any one of items 776 to 868, or a composition as described in item 869 or 870.

[1429] Item 896. A neoantigen vaccine comprising at least one neoantigen mRNA, wherein the at least one neoantigen mRNA encodes a modified or novel neoantigen sequence induced by contacting a proliferative cell with an effective amount of a splicing regulator, an antibody-drug conjugate as described in any one of items 776 to 868, or a composition as described in item 869 or 870.

[1430] Item 897. An antibody-drug conjugate having a structure

[1431]

[1432] Ab is an antibody or antigen-binding fragment that is covalently bound to the maleimide group of ADL1-D1 via the sulfur atom of the thiol group on the antibody or antigen-binding fragment.

[1433] Item 898. An antibody-drug conjugate having a structure

[1434]

[1435] Ab is an antibody or antigen-binding fragment that is covalently bound to the maleimide group of ADL1-D4 via the sulfur atom of the thiol group on the antibody or antigen-binding fragment.

[1436] Item 899. An antibody-drug conjugate having a structure

[1437]

[1438] Ab is an antibody or antigen-binding fragment that is covalently bound to the maleimide group of ADL1-D12 via the sulfur atom of the thiol group on the antibody or antigen-binding fragment.

[1439] Item 900. An antibody-drug conjugate having a structure

[1440]

[1441] Ab is an antibody or antigen-binding fragment that is covalently bound to the maleimide group of ADL5-D2 via the sulfur atom of the thiol group on the antibody or antigen-binding fragment.

[1442] Item 901. An antibody-drug conjugate having a structure

[1443]

[1444] Ab is an antibody or antigen-binding fragment that is covalently bound to the maleimide group of ADL5-D15 via the sulfur atom of the thiol group on the antibody or antigen-binding fragment.

[1445] Item 902. An antibody-drug conjugate having a structure

[1446]

[1447] Ab is an antibody or antigen-binding fragment that is covalently bound to the maleimide group of ADL13-D4 via the sulfur atom of the thiol group on the antibody or antigen-binding fragment.

[1448] Item 903. An antibody-drug conjugate as described in any one of items 897 to 902, wherein the antibody or antigen-binding fragment targets cells expressing HER2.

[1449] Item 904. The antibody-drug conjugate as described in Item 903, wherein the antibody or antigen-binding fragment is an anti-HER2 antibody or antigen-binding fragment.

[1450] Item 905. The antibody-drug conjugate as described in Item 903 or Item 904, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity-determining regions (HCDR1, HCDR2, and HCDR3) containing 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) containing the amino acid sequences of SEQ ID NO:4 (LCDR1), SEQ ID NO:5 (LCDR2), and SEQ ID NO:6 (LCDR3).

[1451] Item 906. The antibody-drug conjugate as described in any one of items 903 to 905, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region containing the amino acid sequence of SEQ ID NO:19 and a light chain variable region containing the amino acid sequence of SEQ ID NO:20.

[1452] Item 907. The antibody-drug conjugate as described in any one of items 903 to 906, wherein the antibody or antigen-binding fragment comprises a constant region of the human IgG1 heavy chain.

[1453] Item 908. An antibody-drug conjugate as described in any one of items 903 to 907, wherein the antibody or antigen-binding fragment comprises a constant region of the human Igκ light chain. Attached Figure Description

[1454] Figure 1 This illustrates the dose response of an exemplary payload compound in a competitive binding assay. Immunoprecipitation was performed on nuclear extracts from 293F cells overexpressing the flag-tagged wild-type SF3B1 using an anti-SF3B1 antibody and a scintillation proximity assay (SPA) bead mixture. The binding reaction contained the antibody-bead mixture and an increased concentration of the compound, followed by [the following text appears to be incomplete and requires further context: "contains anti-SF3B1 antibody and scintillation proximity assay (SPA) bead mixture."] 3 Competition with H-labeled praldadiene lactone B (PB) probe. The y-axis represents the percentage change in specific binding (response %) relative to the DMSO control (0%). Data are expressed as mean ± standard deviation (SD).

[1455] Figure 2This illustrates splicing regulation performed by the exemplary payload compound in an in vitro splicing assay. Nuclear extracts from HeLa S3 cells were incubated with Ad2.2 pre-mRNA and increased concentrations of the compound, followed by quantification of splicing regulation via RT-PCR. The Ad2.2 sequence was derived from an adenoviral Ad2 pre-mRNA substrate with modifications around the branching point sequence. The y-axis represents the percentage change in splicing (response %) relative to the DMSO control (0%). Data are presented as mean ± SD.

[1456] Figure 3 This demonstrates the viability-dose response of an exemplary loaded compound in HER2-amplified breast cancer cells (HCC1954). Cells were incubated with the compound for 144 hours (6 days) and... Viability was read from the reagent. Data are expressed as mean ± SD.

[1457] Figure 4 The results of the cell binding assay are shown. Binding of the exemplary HER2-ADC to JIMT1 cells was assessed by flow cytometry. Mean fluorescence intensity values ​​were measured to determine conjugate binding, followed by determination of binding to the PE-labeled secondary antibody. Data are presented as mean ± SD.

[1458] Figure 5A Viability-dose response of an exemplary HER2-ADC is shown in HER2-amplified breast cancer cells (HCC1954). Figure 5B Viability-dose response of an exemplary HER2-ADC in HER2-amplified gastric cancer cells (N87). Figure 5C Viability-dose response of an exemplary HER2-ADC in HER2-amplified breast cancer cells (SKBR3). Cells were incubated with the conjugate for 144 hours (6 days) and... Viability was read from the reagent. Data are expressed as mean ± SD.

[1459] Figure 6 Viability-dose response of an exemplary HER2-ADC is shown in breast cancer cells (MCF7) that do not express HER2. Cells were incubated with the conjugate for 144 hours (6 days) and Viability was read from the reagent. Data are expressed as mean ± SD.

[1460] Figure 7Results of SLC25A19 splicing assays in HER2-amplified breast cancer cells (HCC1954). Cells were incubated with the conjugate for 24 h, and SLC25A19 transcript splicing was measured using a specific Taqman primer-probe set in a real-time qPCR reaction. The y-axis represents the percentage of response (%) relative to the DMSO control (0%). Data are expressed as mean ± SD.

[1461] Figure 8 The results of the bystander kill assay are shown. H1568 cells overexpressing HER2 (target-positive) or luciferase-labeled H1568 cells (target-negative) were treated with an exemplary HER2-ADC, either plated individually or incubated together in a co-culture for 144 hours (6 days). Reagent reading plate. The y-axis represents the percentage of response (%) relative to the PBS control (100%). Data are presented as mean ± SD.

[1462] Figure 9 This diagram illustrates tumor growth kinetics in each group of CB17-SCID mice (6–10 animals / group) implanted with HCC1954, treated with a single intravenous dose of the exemplary HER2-ADC or a dose-matched payload. Tumor volume was measured twice weekly following treatment. Data are presented as mean ± standard error of mean (SEM).

[1463] Figure 10 Demonstrates the activity-dose response of an exemplary CD138-ADC in a CD138-expressing multiple myeloma cell line. MOLP8 cells were incubated with the conjugate for 144 hours (6 days) and... Viability was read from the reagent. Data are expressed as mean ± SD.

[1464] Figure 11 This demonstrates the activity-dose response of an exemplary EPH2A-ADC in a prostate cancer cell line expressing EPHA2. PC3 cells were incubated with the conjugate for 144 hours (6 days) and... Viability was read from the reagent. Data are expressed as mean ± SD.

[1465] Figure 12A and Figure 12B This shows the results of an in vitro stability assay for an exemplary anti-HER2 ADC, AB185-ADL1-D1. The y-axis represents the total antibody ( Figure 12A ) and the coupled (complete) payload ( Figure 12B The concentration of ) is shown on the x-axis; the x-axis represents the time measured in hours at 37°C.

[1466] Figure 13The plasma concentration of an exemplary anti-HER2 ADC, AB185-ADL1-D1, is shown in CD17-SCID N87 tumor-bearing mice following a single intravenous dose.

[1467] Figure 14 This diagram illustrates an exemplary RNA sequencing and ligandome experiment.

[1468] Figure 15 This diagram illustrates an exemplary T-cell sensitization experiment.

[1469] 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 mixture. FACS was performed to verify the maturation of DCs derived from monocytes.

[1470] Figures 17A-17D This displays the results of the ELISpot measurement. Figure 17A The ELISpot plate was displayed, indicating that neoantigen 1 induced CD8+ T cell activation. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17B Displaying the new antigen 1ELISpot plate ( Figure 17A Quantification of IFNγ spots (number of spots) in IFNγ. Figure 17C The ELISpot plate was displayed, indicating that neoantigen 3 induced CD8+ T cell activation. Stimulation of CD8+ T cells was monitored by IFNγ secretion. Figure 17D Displaying the new antigen 3ELISpot plate ( Figure 17C Quantitative analysis (fold change) of IFNγ spots in ).

[1471] Figure 18 This graph shows a comparison of the splicing power (IC50 qPCR) and cell power (GI50 CTG) of an exemplary anti-HER2 ADC in HCC1954 breast cancer cells. The values ​​shown are sized according to cell lethality and shaded according to the depth of the variable splicing response.

[1472] Figure 19 This graph shows a comparison of the splicing power (IC50 qPCR) and cell power (GI50 CTG) of an exemplary anti-HER2 ADC in N87 gastric cancer cells. The values ​​shown are sized according to cell lethality and shaded according to the depth of the variable splicing response.

[1473] Figure 20This graph shows a comparison of the potency and lethality of an exemplary anti-HER2 ADC (in HCC1954 breast cancer cells) with the stability and permeability of the corresponding payload. The values ​​shown are sized according to payload stability and shaded according to payload permeability.

[1474] Figure 21 This describes tumor growth kinetics in CB17-SCID mice (N=8 mice / group) implanted with N87, treated intravenously for two cycles with a mediator or 10 mg / kg trastuzumab, TDM1, or the exemplary HER2-ADC Q7D. Data are presented as mean ± standard error of mean (SEM) (mm). 3 ).

[1475] Figure 22 This shows the change in body weight in each group of CB17-SCID mice (N=8 mice / group) implanted with N87 after two cycles of intravenous treatment with the medium or 10 mg / kg trastuzumab, TDM1, or the exemplary HER2-ADC Q7D. Data are presented as mean ± standard error of mean (SEM) (%).

[1476] Figure 23 The left panel shows tumor growth kinetics and body weight changes (right panel) in CB17-SCID mice (N=8 mice / group) that were treated intravenously for two cycles with a medium or 10 mg / kg trastuzumab, TDM1, or the exemplary HER2-ADC Q7D. Data are presented as mean ± SEM (tumor volume, mm). 3 (or mean ± SEM (body weight, %))

[1477] Figures 24A-24D This study demonstrates the pharmacodynamic (PD) regulation of mRNA conjugation in N87-implanted CB17-SCID mice treated intravenously for two cycles with a mediator or 10 mg / kg trastuzumab, TDM1, or the exemplary HER2-ADC Q7D. RT-qPCR was used to monitor FBXW5 (mature mRNA transcript) and the results were displayed. Figure 24A and Figure 24C In the middle. RT-qPCR monitoring of TAOK1 (new conjugation transcript) and its display. Figure 24B and Figure 24D In the middle. Within 48 hours ( Figure 24A and Figure 24B ) or at the specified time ( Figure 24C and Figure 24D Animals were collected at a time (N=4 animals / group). Tumors were isolated for RNA extraction and RT-qPCR.

[1478] Figure 25 This diagram illustrates an exemplary target indication analysis.

[1479] Figure 26 An exemplary bioconjugation scheme for preparing ADCs using splicing modulators is shown. Detailed Implementation

[1480] The disclosed compositions and methods can be more readily understood by referring to the following detailed description, which forms part of this disclosure, in conjunction with the accompanying drawings.

[1481] Throughout this disclosure, these descriptions refer to the compositions and methods of using those compositions. When this disclosure describes or claims features or embodiments associated with a composition, such features or embodiments also apply to methods of using the composition. Similarly, when this disclosure describes or claims features or embodiments associated with methods of using the composition, such features or embodiments also apply to the composition.

[1482] When indicating a range of values, it includes embodiments using any particular value within that range. Furthermore, references to values ​​stated within a range include every value within that range. All ranges include their endpoints and are composable. When a value is expressed as an approximation using the antecedent “about,” it will be understood that the particular value forms another embodiment. Unless the context explicitly indicates otherwise, references to a particular numerical value include at least that particular value. Unless otherwise indicated, the use of “or” will mean “and / or.” All references cited herein are incorporated by reference for any purpose. In the event of any conflict between the references and this specification, this specification shall prevail.

[1483] It should be understood that certain features of the disclosed compositions and methods described herein in the context of individual embodiments for clarity may also be provided in combinations of individual embodiments. Conversely, various features of the disclosed compositions and methods described herein in the context of individual embodiments for brevity may also be provided individually or in any sub-combination.

[1484] definition

[1485] Various terms related to the description are used throughout the specification and claims. Unless otherwise specified, such terms will have their ordinary meaning in the art. Other specifically defined terms should be interpreted in accordance with the definitions provided herein.

[1486] As used herein, the singular forms “a / an” and “the” include the plural forms unless the context clearly indicates otherwise.

[1487] As those skilled in the art will recognize from the teachings contained herein, in the context of numerical values ​​and ranges, the term "about / approximately" refers to a value or range that is approximately or close to the stated value or range, such that the embodiment can be performed as intended, such as having the desired amount of nucleic acid or polypeptide in the reaction mixture. In some embodiments, about means a numerical value ±10%.

[1488] The terms “antibody-drug conjugate,” “antibody conjugate,” “conjugate,” “immunoconjugate,” and “ADC” are used interchangeably and refer to one or more therapeutic compounds (e.g., splicing modulators) linked to one or more antibodies or antigen-binding fragments, defined by the following general formula: Ab-(LD) p (Formula I), where Ab = antibody or antigen-binding fragment, L = linker portion, D = drug portion (e.g., splice modulator drug portion), and p = number of drug portions / antibody or antigen-binding fragment. An ADC containing a splice modulator drug portion may also be more specifically referred to herein as an "antibody loaded with splice modulator" or "SMLA". In an ADC containing a splice modulator drug portion, "p" refers to the number of splice modulator compounds attached to the antibody or antigen-binding fragment. In some embodiments, the linker L may include a cleavable portion between the antibody or antigen-binding fragment and the therapeutic compound. In some embodiments, the linker L may include a cleavable portion that can be attached to one or both of the antibody or antigen-binding fragment and the therapeutic compound via one or more spacer subunits. In some embodiments, when the spacer subunit attaches a cleavable portion to the therapeutic compound, it is a self-ablating spacer subunit. In other embodiments, the linker L does not include a cleavable portion and is a non-cleavable linker. In some embodiments, the linker L may include at least one spacer subunit that can be directly attached to the antibody or antigen-binding fragment and the therapeutic compound. Exemplary cleavable and non-cleavable linkers are described and illustrated herein.

[1489] The term "antibody," in its broadest sense, refers to an immunoglobulin molecule that recognizes and specifically binds to a target (such as a protein, polypeptide, carbohydrate, polynucleotide, lipid, or a combination thereof) through at least one antigen recognition site within the variable region of an immunoglobulin molecule. The heavy chain of an antibody consists of a heavy chain variable domain (V... H ) and heavy chain constant region (C H The light chain is composed of light chain variable structural domains (V). L ) and light chain constant structural domain (C LThe mature heavy and light chain variable domains each contain three complementarity-determining regions (CDR1, CDR2, and CDR3) within four framework regions (FR1, FR2, FR3, and FR4) arranged from the N-terminus to the C-terminus: 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 antibodies and full-length polyclonal antibodies, as well as antibody fragments such as Fab, Fab', F(ab')2, Fv, and single-chain antibodies. An antibody can be any of the five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, or a subclass thereof (e.g., isotypes IgG1, IgG2, IgG3, and IgG4). The term further encompasses human antibodies, chimeric antibodies, humanized antibodies, and any modified immunoglobulin molecule containing an antigen recognition site, provided that it exhibits the desired biological activity (e.g., binding to a target antigen, internalization within a cell expressing the target antigen).

[1490] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies, meaning that the individual antibodies constituting the population are identical except for a possible small number of naturally occurring mutations. Monoclonal antibodies are highly specific for a single antigenic epitope. In contrast, conventional (polyclonal) antibody formulations typically comprise multiple antibodies targeting (or specific to) different epitopes. The modifier "monoclonal" indicates the characteristics of an antibody obtained from a substantially homogeneous population of antibodies and should not be construed as requiring the antibody to be produced by any particular method. For example, monoclonal antibodies used according to this disclosure can be prepared by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or by a recombinant DNA method (see, for example, U.S. Patent No. 4,816,567). Monoclonal antibodies can also be isolated from phage antibody libraries using techniques described, for example, those described in Clackson et al. (1991) Nature 352:624-8 and Marks et al. (1991) J. Mol. Biol. 222:581-97.

[1491] The monoclonal antibodies described herein specifically include “chimeric” antibodies, wherein a portion of the heavy chain and / or light chain is identical or homologous to the corresponding sequence of an antibody derived from a particular species or belonging to a particular antibody species or subclass, while the remainder of the one or more chains is identical or homologous to the corresponding sequence in a fragment of an antibody derived from another species or belonging to another antibody species or subclass, provided that they specifically bind to the target antigen and / or exhibit the desired biological activity.

[1492] As used herein, the term "human antibody" refers to an antibody produced by a human or an antibody having the amino acid sequence of an antibody produced by a human.

[1493] As used herein, the term "chimeric antibody" refers to an antibody in which the amino acid sequence of the immunoglobulin molecule is derived from two or more species. In some cases, the variable regions of the heavy and light chains correspond to 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., humans) to minimize the immune response in the latter species.

[1494] As used herein, the term "humanized antibody" refers to an antibody form containing sequences derived from non-human (e.g., mouse) antibodies as well as human antibodies. Such antibodies are chimeric antibodies containing minimal sequences derived from non-human immunoglobulins. Typically, humanized antibodies will contain substantially all, and typically both, variable domains, wherein all or substantially all of the hypervariable loops correspond to those of the non-human immunoglobulin, and all or substantially all of the frame (FR) regions are those regions of the human immunoglobulin sequence. Humanized antibodies may optionally also contain at least a portion of the immunoglobulin constant region (Fc) (typically the human immunoglobulin constant region). Humanized antibodies can be further modified by substitution of residues within the Fv frame regions and / or substituted non-human residues to improve and optimize antibody specificity, affinity, and / or activity.

[1495] As used herein, the term "antigen-binding fragment" or "antigen-binding portion" of an antibody refers to one or more fragments of an antibody or protein that maintain 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). The antigen-binding fragment may also maintain the ability to be internalized into cells expressing the antigen. In some embodiments, the antigen-binding fragment also maintains immune effector activity. It has been shown that fragments of a full-length antibody can perform the antigen-binding function of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding fragment" or "antigen-binding portion" of an antibody include (i) the Fab fragment, a type of antibody composed of V... L V H C L and C H1 (ii) a monovalent segment composed of structural domains; (iii) an F(ab')2 segment, a divalent segment comprising two Fab segments connected by disulfide bonds at the hinge region; H and C H1 (iv) Fd fragments composed of structural domains; V-shaped segments formed by the antibody single armL and V H Fv segments composed of structural domains; (v)dAb segments containing a single variable structural domain, such as V H The structural domains (see, for example, Ward et al. (1989) Nature 341:544-6; and International Publication No. WO 1990 / 005144); and (vi) the separated complementarity-determining regions (CDRs). Furthermore, although the two structural domains V of the Fv segment... L and V H Encoded by independent genes, but which can be joined together using recombination methods via synthetic linkers that enable them to be manufactured as single protein chains, where V L With V H Regions pair to form a monovalent molecule (called a single-chain Fv (scFv)). See, for example, 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" ...

Claims

1. An antibody-drug conjugate of formula (I), Ab-(L-D) p (I) in Ab is an antibody or antigen-binding fragment that targets neoplastic cells; D is a splicing regulator; L is a linker that covalently links Ab to D; and p is an integer of 1-15.

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

4. The antibody-drug conjugate of claim 3, wherein the hematological malignancy is selected from the group consisting of B-cell malignancies, leukemias, lymphomas, and myelomas.

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

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

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 carcinoma, 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 cells expressing HER2.

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 claim 9, wherein the antibody or antigen-binding fragment comprises three heavy chain complementary 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 complementary 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).

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