Antibody-drug conjugates containing NMT inhibitors and uses thereof

Abstract

The present invention relates to ADCs comprising NMT inhibitors conjugated to antibodies via linkers, and related uses.

Description

[Technical Field]

[0001] The present invention relates to novel antibody drug conjugates (ADCs) comprising inhibitors of human N-myristoyltransferase (human NMT). The present invention also relates to such ADCs for use as pharmaceuticals, particularly in the treatment or prevention of hyperproliferative disorders such as cancer. [Background technology]

[0002] Antibody therapy has been established for the targeted treatment of patients with cancer, immunological disorders, and angiogenic disorders (Carter, P (2006) Nature Reviews Immunology 6:343-357). While the use of antibody-drug conjugates (ADCs), i.e., immunoconjugates, for the local delivery of cytotoxic or cytostatic agents, i.e., drugs that kill or inhibit tumor cells, in the treatment of cancer targets the delivery of the drug moiety to the tumor and its intracellular accumulation therein, systemic administration of these unconjugated drug substances can result in unacceptable levels of toxicity to normal cells (Xie et al. (2006) Expert. Opin. Biol. Ther. 6(3):281-291; Kovtun et al. (2006) Cancer Res. 66(6):3214-3121; Law et al. (2006) Cancer Res. 66(4):2328-2337; Wu et al. (2005) Nature Biotech. 23(9):1137-1145; Lambert J. (2005) Current Opin. in Pharmacol. 5:543-549; Hamann P (2005) Expert Opin. Ther. Patents 15(9):1087-1103; Payne, G (2003) Cancer Cell 3:207-212; Trail et al. (2003) Cancer Immunol. Immunother. 52:328-337; Syrigos and Epenetos (1999) Anticancer Research 19:605-614).

[0003] Maximum efficacy with minimal toxicity is thereby sought. Efforts to design and refine ADCs have focused on monoclonal antibody (mAb) selectivity, as well as drug mechanism of action, drug binding, drug / antibody ratio (loading), and drug release characteristics (Junutula et al., 2008b Nature Biotech., 26(8):925-932; Dornan et al., (2009) Blood 114(13):2721-2729; US 7521541; US ​​7723485; WO2009 / 052249; McDonagh, (2006) Protein Eng. Design & Sei. 19(7):299-307; Doronina, et al., (2006) Bioconj. Chem. 17:114-124; Erickson, et al., (2006) Cancer Res. 66(8):1-8; Sanderson et al. (2005) Clin. Cancer Res. 11:843-852; Jeffrey et al. (2005) J. Med. Chem. 48:1344-1358; Hamblett et al. (2004) Clin. Cancer Res. 10:7063-7070). Drug moieties can confer their cytotoxic and cytostatic effects by mechanisms including tubulin binding, DNA binding, proteasome, and / or topoisomerase inhibition. Some cytotoxic drugs tend to become inactive or less active when conjugated to large antibodies or protein receptor ligands.

[0004] N-myristoyltransferase (NMT) is a monomeric enzyme ubiquitous in eukaryotes that catalyzes the irreversible cotranslational transfer of myristate (a saturated 14-carbon fatty acid) from myristoyl-coenzyme A (myr-CoA) to protein substrates containing N-terminal glycines, accompanied by the formation of an amide bond (Farazi, T.A., G. Waksman, and J.I. Gordon, J. Biol. Chem., 2001, 276(43):39501-39504).

[0005] There are two human NMTs: human NMT1 (HsNMT1) and human NMT2 (HsNMT2).Inhibition of human NMTs has been shown to be effective in treating a variety of diseases or disorders, such as hyperproliferative disorders (e.g., cancers, e.g., human colorectal cancer, gallbladder cancer, brain tumors, and lymphomas, e.g., B-cell lymphomas) (Resh MD, 1993. Biochern. Biophys. Acta 1115, 307-22; Bertiaume LG, Beuachamp E, WO2017011907), as well as viral infections, such as HIV (Gottlinger HG, Sodroski JG, Haseltine WA, 1989. Proc. Nat. Acad. Sci. USA 86:5781-85; Bryant ML, Ratner L, 1990. Proc. Natl. Acad. Sci. USA 87:523-27) and human rhinovirus (HRV) (Davis MP, Bottley G, Beales RD, 1994. Proc. Natl. Acad. Sci. USA 87:523-27). References LP, Killington, RA, Rowlands DJ, Tuthill, TJ, 2008 Journal of Virology 82 4169-4174;Mousnier A, Bell AS, Swieboda DP, Morales-Sanfrutos J, Perez-Dorado I, Brannigan JA, Newman J, Ritzefeld M, Hutton, JA, Guedan A, Asfor AS, Robinson, SW, References Hopkins-Navratilova I, Wilkinson AJ, Johnston SL, Leatherbarrow RJ, Tuthill TJ, Solari R, Tate EW, 2018 Nature Chemistry 10(6) 599-606), Corbic Ramljak I, Stanger J, Real-Hohn A. Dreier D, Wimmer L., Redlberger-Fritz M, Fischl W, Klingel K, Mihovilovic MD, Blaas D, Kowalski H, PLOS Pathogens 14(8):e1007203) have been proposed as targets for treating or preventing cancer.Because NMTs play important roles in protein trafficking, mediating protein-protein interactions, stabilizing protein structure, and signal transduction in biological systems, inhibition of HsNMT1 and / or HsNMT2 enzymes has the potential to disrupt multiprotein pathways. While inhibition of human NMTs is thought to inhibit both HsNMT1 and HsNMT2, its therapeutic and / or prophylactic activity is thought to derive primarily from inhibition of HsNMT1. The above characteristics are thought to be desirable, for example, to reduce the risk of resistance development in the treatment or prevention of microbial infections and hyperproliferative disorders.

[0006] NMT has two known binding pockets: the myr-CoA binding pocket and the peptide binding pocket. Most NMT inhibitors reported to date target the peptide binding pocket.

[0007] Compounds active as inhibitors of NMT have been previously disclosed, see, for example, WO00 / 37464 (Roche), WO2010 / 026365 (University of Dundee), WO2013 / 083991 (Imperial Innovations Limited), WO2017 / 001812 (Imperial Innovations Limited), WO2020 / 128473 (Imperial College Innovations Limited), WO2020 / 128475 (Imperial College Innovations Limited), and WO2022 / 058745 (Imperial College Innovations Limited et al.). Certain uses of NMT inhibitors have been disclosed, see for example WO2022 / 090746 (Imperial College Innovations Limited et al.) and WO2022 / 082306 (Pacylex Pharmaceuticals).

[0008] However, there remains a need for additional ADCs that retain the efficacy of the conjugated drug moiety while controlling toxicity.

[0009] Surprisingly, the present inventors have now discovered that ADCs comprising NMT inhibitors exhibit potent cytotoxic activity with minimal adverse side effects such as weight loss. These properties are believed to make the ADCs of the present invention particularly suitable for use as pharmaceuticals for the treatment or prevention of hyperproliferative diseases such as cancer. Summary of the Invention

[0010] (Summary of the Invention) The present invention provides antibody drug conjugates (ADCs) or salts thereof comprising an NMT inhibitor conjugated to an antibody via a linker (herein referred to as "ADCs of the invention").

[0011] The ADC of the present invention may be provided in the form of a salt. Preferably, the ADC of the present invention is provided in the form of a pharmaceutically acceptable salt. Preferably, the ADC of the present invention is provided as follows:

[0012] The present invention also provides pharmaceutical compositions comprising an ADC of the invention or a pharmaceutically acceptable salt thereof.

[0013] The present invention also provides an ADC of the invention or a pharmaceutically acceptable salt thereof for use as a pharmaceutical.

[0014] The present invention also provides an ADC of the invention, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a disease or disorder (e.g., cancer) for which inhibition of N-myristoyltransferase provides a therapeutic or preventative effect.

[0015] The invention also provides use of an ADC of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment or prevention of a disease or disorder (e.g., cancer) for which inhibition of N-myristoyltransferase provides a therapeutic or prophylactic effect.

[0016] The present invention also provides methods for preventing or treating diseases or disorders (e.g., cancer) for which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect, the methods comprising administering a therapeutically effective amount of an ADC of the invention or a pharmaceutically acceptable salt thereof. [Brief explanation of the drawings]

[0017] DESCRIPTION OF THE DRAWINGS [Figure 1] FIG. 1: Shows the effect of treatment with trastuzumab (2.5 mg / kg), ADC Example 1 (2.5 mg / kg), or NMT inhibitor 1 (2 mg / kg) on ​​tumor volume in a mouse xenograft study. [Figure 2] FIG. 2: Shows the effect of treatment with trastuzumab (5 mg / kg), ADC Example 1 (5 mg / kg), or NMT inhibitor 1 (2 mg / kg) on ​​tumor volume in a mouse xenograft study. [Figure 3] FIG. 3: Shows the effect of treatment with trastuzumab (2.5 mg / kg), ADC Example 1 (2.5 mg / kg), or NMT inhibitor 1 (2 mg / kg) on ​​mouse body weight in a mouse xenograft study. [Figure 4] FIG. 4: Shows the effect of treatment with trastuzumab (5 mg / kg), ADC Example 1 (5 mg / kg), or NMT inhibitor 1 (2 mg / kg) on ​​mouse body weight in a mouse xenograft study. [Figure 5] Figure 5: Shows the effect of treatment with 2.5 mg / Kg trastuzumab, 2.5 mg / Kg ADC Example 1, 2.5 mg / Kg trastuzumab deruxtecan, and 5 mg / Kg isotype control antibody on tumor volume in a mouse gastric cancer xenograft model. [Figure 6] Figure 6: Shows the effect of treatment with 5 mg / Kg trastuzumab, 5 mg / Kg ADC Example 1, 5 mg / Kg trastuzumab deruxtecan, and 5 mg / Kg isotype control antibody on tumor volume in a mouse gastric cancer xenograft model. [Figure 7]Figure 7A: shows the % weight change in mice following the experiment described in Biological Example 3 (and Figure 5, 2.5 mg / kg (mpk)). Figure 7B: shows the % weight change in mice following the experiment described in Biological Example 3 (and Figure 6; 5 mg / kg (mpk)). [Figure 8] Figure 8: Shows the results of in vitro evaluation of the cytotoxicity of trastuzumab conjugated to NMT inhibitor 1 (ADC Example 1; Figure 8) against the HER2-positive breast cancer cell line BT474 at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM). Puromycin was used as a positive cytotoxicity control. [Figure 9] Figure 9: Results of in vitro evaluation of cytotoxicity of trastuzumab (Figure 9) against the HER2-positive breast cancer cell line BT474 at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM). Puromycin was used as a positive cytotoxicity control. [Figure 10] Figure 10: Results of in vitro evaluation of the cytotoxicity of trastuzumab deruxtecan (Figure 10) at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM) against the HER2-positive breast cancer cell line BT474. Puromycin was used as a positive cytotoxicity control. [Figure 11] Figure 11: Shows the results of in vitro evaluation of the cytotoxicity of an isotype control (isotype control IgG conjugated to NMT inhibitor 1; Figure 11) at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM) against the HER2-positive breast cancer cell line BT474. Puromycin was used as a positive cytotoxicity control. [Figure 12] Figure 12: Shows the results of in vitro evaluation of the cytotoxicity of trastuzumab conjugated to NMT inhibitor 1 (ADC Example 1; Figure 12) at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM) against the HER2-negative breast cancer cell line MCF7. Puromycin was used as a positive cytotoxicity control. [Figure 13]Figure 13: Shows the results of in vitro evaluation of the cytotoxicity of trastuzumab (Figure 13) at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM) against the HER2-negative breast cancer cell line MCF7. Puromycin was used as a positive cytotoxicity control. [Figure 14] Figure 14: Shows the results of in vitro evaluation of the cytotoxicity of trastuzumab deruxtecan (Figure 14) at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM) against the HER2-negative breast cancer cell line MCF7. Puromycin was used as a positive cytotoxicity control. [Figure 15] Figure 15: Shows the results of in vitro evaluation of the cytotoxicity of an isotype control (isotype control IgG conjugated to NMT inhibitor 1; Figure 15) at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM) against the HER2-negative breast cancer cell line MCF7. Puromycin was used as a positive cytotoxicity control. [Figure 16] Figure 16: Shows the in vitro "bystander effect" results for trastuzumab conjugated to NMT inhibitor 1 (ADC Example 1), trastuzumab, Reference Example 1 (trastuzumab conjugated to monomethyl auristatin E (MMAE)), isotype control, and trastuzumab deruxtecan at different concentrations (12.5 nM, 3.13 nM, 0.78 nM, 0.2 nM, and 0.05 nM) on the HER2-negative breast cancer cell line MCF7. [Figure 17] Figure 17: Shows the in vitro "bystander effect" results for trastuzumab conjugated to NMT inhibitor 1 (ADC Example 1), trastuzumab, Reference Example 1 (trastuzumab conjugated to monomethyl auristatin E (MMAE)), isotype control, and trastuzumab deruxtecan at different concentrations (12.5 nM, 3.13 nM, 0.78 nM, 0.2 nM, and 0.05 nM) on the HER2-positive breast cancer cell line BT474. [Figure 18]Figure 18: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including BT474 (Figure 18), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 19] Figure 19: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including JIMT 1 (Figure 19), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 20] Figure 20: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including NCI N87 (Figure 20), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 21] Figure 21: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including NCI H292 (Figure 21), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 22] Figure 22: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including IM95-m (Figure 22), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 23] Figure 23: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including ZR-75-30 (Figure 23), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 24]Figure 24: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including NCI H2170 (Figure 24), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 25] Figure 25: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including LNCaP (Figure 25), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 26] Figure 26: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including C42 (Figure 26), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 27] Figure 27: Shows the results of in vitro evaluation of the cytotoxicity of ADC Examples 1-5 against a panel of cell lines, including VCaP (Figure 27), using various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) provided in each figure. [Figure 28] Figure 28: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA0429B (Figure 28) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 29] Figure 29: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA6877B (Figure 29) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 30]Figure 30: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA6894B (Figure 30) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 31] Figure 31: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA2434B (Figure 31) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 32] Figure 32: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA3102B (Figure 32) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 33] Figure 33: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA0119B (Figure 33) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 34] Figure 34: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA0091B (Figure 34) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 35]Figure 35: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA6815B (Figure 35) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 36] Figure 36: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA0098B (Figure 36) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 37] Figure 37: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA6833B (Figure 37) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 38] Figure 38: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA0087B (Figure 38) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 39] Figure 39: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA2109B (Figure 39) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 40]Figure 40: Shows the results of the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA3055B (Figure 40) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 41] Figure 41: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA6866B (Figure 41) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 42] Figure 42: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA0060B (Figure 42) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 43] Figure 43: Results show the effect of test articles isotype control, ADC Example 1, staurosporine, trastuzumab, and trastuzumab deruxtecan as single agents on cell viability of the following gastric cancer organoid line: GA6891B (Figure 43) using a CellTiter-Glo (CTG) luminescent cell viability assay. [Figure 44] Figure 44a-b: Shows the mean (+ / - SEM) levels of hematological markers when normalized to baseline (pre-dose) values ​​in the 20 mg / Kg group described in Biological Example 9. [Figure 45] FIG. 45: Shows weight loss in mice in Biological Example 10. [Figure 46]Figure 46: Shown from left to right are the mean values ​​per dose of AST (aspartate transaminase), ALT (alanine transaminase), ALP (alkaline phosphatase), LDH (lactate dehydrogenase), CK (creatine kinase), and GGT (gamma-glutamyltransferase) in the blood of animals at the time of slaughter in Biological Example 10. [Figure 47] Figure 47: Mean values ​​per dose of LYM, MON, NEU, RBC, HGB, and PLT cells in the blood of animals at the time of sacrifice in Biological Example 10 are shown from left to right. [Figure 48] Figure 48: Shows the effect of treatment with ifinatamab (5 mg / kg and 10 mg / kg), ifinatamab-DXd (5 mg / kg and 10 mg / kg), ADC Example 4 (5 mg / kg and 10 mg / kg), and vehicle control on tumor volume in a mouse LNCaP prostate cancer xenograft model. [Figure 49] Figure 49: Shows the effect of treatment with ifinatamab (5 mg / kg and 10 mg / kg), ifinatamab-DXd (5 mg / kg and 10 mg / kg), ADC Example 4 (5 mg / kg and 10 mg / kg), and vehicle control on body weight in a mouse LNCaP prostate cancer xenograft model. [Figure 50] Figure 50: Shows the effect of treatment with ifinatamab (5 mg / kg), ifinatamab-DXd (5 mg / kg), ADC Example 4 (2.5 mg / kg, 5 mg / kg, and 10 mg / kg), and vehicle control on tumor volume in a mouse VCaP prostate cancer xenograft model. [Figure 51] Figure 51: Shows the effect of treatment with ifinatamab (5 mg / kg), ifinatamab-DXd (5 mg / kg), ADC Example 4 (2.5 mg / kg, 5 mg / kg, and 10 mg / kg), and vehicle control on body weight in a mouse VCaP prostate cancer xenograft model. [Figure 52]Figure 52: Shows the effect of treatment with sacituzumab (5 mg / kg), sacituzumab govitecan (5 mg / kg) (plus 5 mg / kg of ADC Example 3 added on study days 27 and 34), and ADC Example 3 (5 mg / kg), and vehicle control on tumor volume in a murine JIMT-1 breast cancer xenograft model. [Figure 53] Figure 53: Shows the effect of treatment with sacituzumab (2.5 mg / kg), sacituzumab govitecan (2.5 mg / kg plus 5 mg / kg of ADC Example 3 added on study day 27), and ADC Example 3 (2.5 mg / kg), and vehicle control on tumor volume in a murine JIMT-1 breast cancer xenograft model. [Figure 54] Figure 54: Shows the effect of treatment with sacituzumab (5 mg / kg), sacituzumab govitecan (5 mg / kg) (plus 5 mg / kg of ADC Example 3 added on study days 27 and 34), and ADC Example 3 (5 mg / kg and 10 mg / kg), and vehicle control on body weight in a murine JIMT-1 breast cancer xenograft model. [Figure 55] Figure 55: Shows the effect of treatment with sacituzumab (2.5 mg / kg), sacituzumab govitecan (2.5 mg / kg plus 5 mg / kg of ADC Example 3 added on study day 27), and ADC Example 3 (2.5 mg / kg), and vehicle control on body weight in a murine JIMT-1 breast cancer xenograft model. [Figure 56] Figure 56 shows the results of in vitro evaluation of the cytotoxicity of trastuzumab conjugated to NMT inhibitor 1 (ADC Example 8) at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM) against the HER2-positive breast cancer cell line BT474. Puromycin was used as a positive cytotoxicity control. [Figure 57] Figure 57 shows the results of in vitro evaluation of the cytotoxicity of trastuzumab conjugated to NMT inhibitor 1 (ADC Example 8) at different concentrations (50 nM, 3.13 nM, 0.2 nM, and 0 nM) against the HER2-negative breast cancer cell line MCF7. Puromycin was used as a positive cytotoxicity control. DETAILED DESCRIPTION OF THE INVENTION

[0018] (Sequence Listing) SEQ ID NO: 1 - Amino acid sequence of the light chain of trastuzumab SEQ ID NO:2 - Amino acid sequence of the heavy chain of trastuzumab SEQ ID NO: 3 - Amino acid sequence of the light chain of rituximab SEQ ID NO: 4 - Amino acid sequence of the heavy chain of rituximab SEQ ID NO: 5 - Amino acid sequence of the light chain of ifinatamab SEQ ID NO: 6 - Amino acid sequence of the heavy chain of ifinatamab SEQ ID NO: 7 - Amino acid sequence of the light chain of sacituzumab SEQ ID NO: 8 - Amino acid sequence of the heavy chain of sacituzumab

[0019] (Detailed Description of the Invention) The term "hydrocarbyl," as used herein, is understood to mean any compound of a straight-chain or branched-chain saturated, unsaturated, or partially unsaturated hydrocarbon group. Suitable examples of "hydrocarbyl" groups can include, for example, "alkyl," "alkenyl," "alkynyl," and / or "haloalkyl" groups, each of which is as defined below.

[0020] The term "alkyl" as used herein is understood to mean straight-chain and branched saturated hydrocarbon groups. Examples of "alkyl" groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, i-butyl, sec-butyl, pentyl, and hexyl groups. Among unbranched alkyl groups, methyl, ethyl, n-propyl, isopropyl, and n-butyl groups are preferred. Among branched alkyl groups, t-butyl, i-butyl, 1-ethylpropyl, and 1-ethylbutyl groups can be mentioned.

[0021] "C" used alone or as a prefix m-n The term "(m-nC) group" or "(m-nC) group" refers to any group having m to n carbon atoms.

[0022] As used herein, the term "alkenyl" refers to both straight-chain and branched-chain unsaturated hydrocarbon groups having at least one carbon-carbon double bond. Examples of alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, and hexenyl. Preferred alkenyl groups include ethenyl, 1-propenyl, 2-propenyl, and but-2-enyl.

[0023] As used herein, the term "alkynyl" refers to both straight-chain and branched-chain unsaturated hydrocarbon groups having at least one carbon-carbon triple bond. Examples of alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, and hexynyl. Preferred alkynyl groups include ethynyl, 1-propynyl, and 2-propynyl.

[0024] The term "carbocyclyl" (or "carbocycle"), as used herein, is intended to mean any 3- to 13-membered carbon ring system, which may be saturated, partially unsaturated, or aromatic. The carbocyclic ring system may be monocyclic or contain multiple rings (e.g., the ring system may be bicyclic). Examples of monocyclic saturated carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of bicyclic saturated carbocycles include bicyclooctane, bicyclononane, bicyclodecane (decalin), and bicyclooctane. A further example of a saturated carbocycle is adamantane. Examples of monocyclic unsaturated carbocycles include cyclobutene, cyclopentene, cyclopentadiene, and cyclohexene. Examples of aromatic carbocycles include phenyl and naphthyl. Further examples of aromatic carbocycles include tetrahydronaphthyl (tetralin) and indane.

[0025] The term "cycloalkyl" as used herein refers to a saturated group in a ring system. Cycloalkyl groups can be monocyclic or bicyclic. Bicyclic groups can be, for example, fused or bridged. Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, and cyclopentyl. Other examples of monocyclic cycloalkyl groups are cyclohexyl, cycloheptyl, and cyclooctyl. Examples of bicyclic cycloalkyl groups include bicyclo[2.2.1]hept-2-yl. Preferably, the cycloalkyl group is monocyclic.

[0026] As used herein, the term "halogen" or "halo" means fluorine, chlorine, bromine, or iodine, with fluorine, chlorine, and bromine being particularly preferred.

[0027] As used herein, the term "haloalkyl" refers to an alkyl group having a halogen substituent, and the terms "alkyl" and "halogen" are understood to have the meanings outlined above. Similarly, the term "dihaloalkyl" refers to an alkyl group having two halogen substituents, and the term "trihaloalkyl" refers to an alkyl group having three halogen substituents. Examples of haloalkyl groups include fluoromethyl, chloromethyl, bromomethyl, fluoromethyl, fluoropropyl, and fluorobutyl groups; examples of dihaloalkyl groups include difluoromethyl and difluoroethyl groups; examples of trihaloalkyl groups include trifluoromethyl and trifluoroethyl groups.

[0028] The term "heterocyclyl" (or heterocycle), as used herein, means an aromatic or non-aromatic cyclic group of carbon atoms, wherein 1 to 4 of the carbon atoms are replaced by one or more heteroatoms independently selected from nitrogen, oxygen, or sulfur. A heterocyclyl (or heterocycle) group may be, for example, monocyclic or bicyclic. In a bicyclic heterocyclyl (or heterocycle) group, one or more heteroatoms may be in each ring or in only one of the rings. The heteroatoms may be S, O, or N, and are preferably O or N.

[0029] Examples of monocyclic non-aromatic heterocyclyls (or heterocycles) include aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, and azepanyl.

[0030] Examples of monocyclic aromatic heterocyclyl (or heterocycle) groups include furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, oxadiazolyl, thiadiazolyl, pyridyl, triazolyl, triazinyl, tetrazolyl, pyridazyl, isothiazolyl, isoxazolyl, pyrazinyl, pyrazolyl, and pyrimidinyl.

[0031] Examples of bicyclic aromatic heterocyclyl groups (or heterocycles) include quinoxalinyl, quinazolinyl, pyridopyrazinyl, benzoxazolyl, benzothiophenyl, benzimidazolyl, naphthyridinyl, quinolinyl, benzofuranyl, indolyl, benzothiazolyl, oxazolyl[4,5-b]pyridiyl, pyridopyrimidinyl, isoquinolinyl, and benzodroxazole. Further examples of bicyclic aromatic heterocyclyl groups include those in which one ring is aromatic and the other is non-aromatic, such as dihydrobenzofuranyl, indanyl, indolinyl, isoindolinyl, tetrahydroisoquinolinyl, tetrahydroquinolyl, and benzazepanyl.

[0032] The term "optionally substituted" refers to groups, structures, or molecules that are substituted and to groups, structures, or molecules that are not substituted. 1 The term "one / any CH, CH, CH group, or heteroatom (i.e., NH) within a group is optionally substituted" preferably refers to R 1 It means that one (any) of the hydrogen radicals of the group is replaced by the associated defined group.

[0033] Where any substituent is selected from "one or more" groups, this definition should be understood to include all substituents selected from one of the specified groups or substituents selected from two or more of the specified groups.

[0034] (NMT inhibitor) As outlined above, the ADCs of the present invention comprise an NMT inhibitor. In one embodiment, the NMT inhibitor is a compound of formula (I): [ka] (In the formula: Y is -CH-, -C(R 2 )-, and -N-; R 1 is a group of formula -XLA; X represents -O-; L is -(CH2) m - represents; m is 1, 2, or 3; A is a 6- to 10-membered aromatic carbocycle or a 5- to 10-membered aromatic heterocycle, and the aromatic carbocycle or heterocycle is not substituted with -F, -Cl, -Br, -OCH3, -OCF3, -CN, up to three halogens, hydroxyl, or -OC 1-4 -C optionally substituted with alkyl groups 1-6 Alkyl, -S(O)C 1-4 Alkyl, -S(O)2C 1-4 Alkyl, -C(O)N(R 9)2, -C(O)N(R 13 )C 1-4 Alkyl OC 1-4 Alkyl, -C(O)N(C 1-4 Alkyl OC 1-4 alkyl), -CHC(O)N(R 9 )2, -CH2C(O)N(R 13 )C 1-4 Alkyl OC 1-4 Alkyl, -CHC(O)N(C 1-4 Alkyl OC 1-4 alkyl)2, -S(O)2NHC 1-4 Alkyl, -S(O)N(C 1-4 alkyl)2, -NHC 1-4 Alkyl, -N(C 1-4 alkyl)2, -NHC(O)C 1-4 Alkyl, -NHC(O)CF3, -NHS(O)2C 1-4 Alkyl, CHN(R 13 )2, CH2N(R 13 )C(O)C 1-4 Alkyl, CHN(R 13 )S(O)2C 1-4 Alkyl, -CH2S(O)2C 1-4 optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of alkyl, and COH; s is 0, 1, 2, or 3; Each R 2 is -F, -Cl, -Br, -OCH3, -OCF3, -CN, -C optionally substituted by up to three halogen or hydroxyl groups 1-4 Alkyl, -S(O)C 1-4 Alkyl, -S(O)2C 1-4 Alkyl, -S(O)2NHC 1-4 Alkyl, -S(O)N(C 1-4 alkyl)2, -NHC 1-4 Alkyl, -N(C 1-4 alkyl)2, -NHC(O)C 1-4 Alkyl, -NHC(O)CF3, and -NHS(O)2C 1-4 independently selected from the group consisting of alkyl; q is 0 or 1; R 3 is hydrogen or methyl; R 4 is hydrogen or methyl; R 5 is hydrogen; R 6 is a C optionally substituted with hydrogen or up to three -F, -Cl, -Br, -OH, -OCH3, -OCF3, or -CN groups 1-6 is alkyl; If present, R 10 is hydrogen or methyl; If present, R 11 is hydrogen or methyl; or R 3 Groups and R 5 The group and the intervening atom may be an intervening atom and a bond, or an intervening atom and -(CHR a ) r -, or R 10 Groups and R 5 The group and the intervening atom are represented by the intervening atom and -(CHR a ) r -, forming a 3- to 7-membered non-aromatic heterocycle; r is 1, 2, 3, 4, or 5; R a is hydrogen or methyl; Each R 7 is hydrogen, halogen, C 1-4 C optionally substituted with alkoxy and one, two, or three halogens 1-4 independently selected from the group consisting of alkyl; and R 8 is hydrogen and C 1-4 alkyl; Each R 9 is hydrogen and C 1-4 alkyl, or two R 9 the groups and the N to which they are attached form a 4- to 7-membered non-aromatic heterocycle, which optionally contains one or two additional heteroatoms selected from N, O, and S; Each R 13 is hydrogen and C 1-4alkyl; and wherein i) E, J, and G are each C(R 7 ), K is carbon, and Q is N(R 8 ) and M is nitrogen; ii) E, J, and G are each C(R 7 ) and K, Q, and M are each nitrogen; or iii) E, J, G, and M are each C(R 7 ) and K and Q are each nitrogen.

[0035] Suitably, the compound of formula (I) or a salt thereof is N(R 5 )(R 6 ) group to the linker.

[0036] In one embodiment, the NMT inhibitor is a compound of formula (IA^^): or a salt thereof [ka] (In the formula: R 1 is a group of formula -XLA; A is 4-pyrazolyl, wherein the pyrazolyl is optionally substituted with up to three substituents selected from methyl and —C(O)N(CH) ; X is -O-; L is -(CH2) m - and; m is 2; R 2' is selected from the group consisting of fluorine or chlorine (preferably fluorine); R 2'' is selected from the group consisting of hydrogen, fluorine, or chlorine; q is 0; R 3 is hydrogen or methyl; R 4 is hydrogen or methyl; R 5 is hydrogen; R 6is hydrogen or methyl; or R 3 Groups and R 6 The group and the intervening atom(s) form a 3- to 7-membered non-aromatic heterocycle consisting of the intervening atom(s) and the bond; E, J, G, K, Q, and M are: i) E, J, and G are each CH, K is carbon, and Q is N(R 8 ), M is nitrogen; and R 8 is hydrogen or methyl; or ii) E, J, G, and M are each CH and K and Q are each nitrogen; provided that A is substituted with up to one -C(O)N(CH3)2 group).

[0037] Preferably, the compound of formula (IA^^) or a salt thereof is N(R 5 )(R 6 ) group to the linker.

[0038] Compounds of formula (I) and (IA^^) are disclosed in WO2017 / 001812, the entire contents of which are incorporated by reference for purposes of describing the synthesis and activity of NMT inhibitors.

[0039] In one embodiment, the NMT inhibitor is a compound of formula (II): [ka] (In the formula: R 1 is H or -CH3; and R 2 is H or F).

[0040] Preferably, the compound of formula (II) or a salt thereof is attached to the linker via the NH2 group.

[0041] In one embodiment, the NMT inhibitor is 4-(2-{2-[3-(2-aminoethyl)imidazo[1,2-a]pyridin-6-yl]-5-chlorophenoxy}ethyl)-N,N,1,5-tetramethyl-1H-pyrazole-3-carboxamide: [ka] or a salt thereof.

[0042] Preferably, the compound 4-(2-{2-[3-(2-aminoethyl)imidazo[1,2-a]pyridin-6-yl]-5-chlorophenoxy}ethyl)-N,N,1,5-tetramethyl-1H-pyrazole-3-carboxamide or a salt thereof is attached to the linker via the NH group.

[0043] The compound of formula (II) and 4-(2-{2-[3-(2-aminoethyl)imidazo[1,2-a]pyridin-6-yl]-5-chlorophenoxy}ethyl)-N,N,1,5-tetramethyl-1H-pyrazole-3-carboxamide are disclosed in WO2020 / 128473, the entire contents of which are incorporated by reference for purposes of describing the synthesis and activity of NMT inhibitors.

[0044] In one embodiment, the NMT inhibitor is a compound of formula (III) or (IV): [ka] (In the formula: n1 is 0, 1, 2, 3, 4, 5, or 6; Ring A * is an optionally substituted nitrogen-containing aryl group, where ring A * Each substitutable carbon or nitrogen in 5A and wherein ring A is optionally and independently substituted by * contains an -NH- moiety, the nitrogen is C 1-6 optionally substituted with alkyl (e.g., methyl); and wherein R 4A and Ring A *may together with the atoms to which they are attached form a cyclic group; Ring B * is an optionally substituted aryl or heteroaryl group, wherein ring B * Each substitutable carbon or heteroatom in 3A optionally and independently substituted by; One of W and X may be absent. 11A , R 11A optionally substituted hydrocarbyl (e.g., C 1-8 alkyl, alkenyl, alkynyl, or haloalkyl), and R 12A optionally substituted with -(CH2) k1 -heterocyclyl; k1 is 0, 1, 2, 3, 4, 5, or 6; R 1A is hydrogen; R 2A , R 3A , R 4A , and R 5A is hydrogen, R 12A , R 12A optionally substituted hydrocarbyl (e.g., C 1-6 alkyl, alkenyl, alkynyl, or haloalkyl), and one or more R 12A optionally substituted with -(CH2) L1 -heterocyclyl; where R 2A together with W or X, one or more R 12A may form an optionally substituted heterocycle; and wherein R 3A and R 5A One or more of, together with the atom to which they are attached, R 12A may form an optionally substituted carbocycle, e.g., heterocyclyl; L1 is 0, 1, 2, 3, 4, 5, or 6; where: Each R 11A and R 12A is halogen, trifluoromethyl, cyano, thio, nitro, oxo, =NR 13A , -OR 13A , -SR 13A, -C(O)R 13A , -C(O)OR 13A , -OC(O)R 13A , -NR 13A COR 14A , -NR 13A CON(R 13A )2, -NR 13a COR 14a , -NR 13a CO2R 14A , -S(O)R 13A , -S(O)2R 13A , -SON(R 13A )2, -NR 13A S(O)2R 14A ;-CSR 13A , -N(R 13A )R 14A , -C(O)N(R 13A )R 14A , -SO2N(R 13A )R 14A , and R 15A are independently selected from; R 13A and R 14A is hydrogen or R 15A are each independently selected from R 15A is a hydrocarbyl (e.g., C 1-6 alkyl, alkenyl, alkynyl, or haloalkyl), carbocyclyl, and -(CH) m1 -heterocyclyl, and each R 15A is halogen, cyano, amino, hydroxy, C 1-6 Alkyl or cycloalkyl, and C 1-6 optionally and independently substituted with one or more of alkoxy; m1 is 0, 1, 2, 3, 4, 5, or 6; p1 is 0, 1, 2, 3, or 4; R 4A The values ​​of may be the same or different; and q1 is 0, 1, 2, 3, or 4; where R 5A The values ​​of may be the same or different; Y and Z, one or both of which may be absent, are hydrogen, R 16A, R 16A optionally substituted hydrocarbyl (e.g., C 1-6 alkyl, alkenyl, alkynyl, or haloalkyl), and R 16A optionally substituted with -(CH2) r1 -heterocyclyl, wherein each R 16A is halogen, trifluoromethyl, cyano, thio, nitro, oxo, =NR 17A , -OR 17A , -SR 17A , -C(O)R 17A , -C(O)OR 17A , -OC(O)R 17A , -NR 17A COR 18A , -NR 17A CON(R 18A )2, -NR 17A COR 18A , -NR 17A CO2R 18A , -S(O)R 17A , -S(O)2R 17A , -SON(R 17A )2, -NR 17A S(O)2R 18A ;-CSR 17A , -N(R 17A )R 18A , -C(O)N(R 17A )R 18A , -SO2N(R 17A )R 18A , and R 19A r1 is 0, 1, 2, 3, 4, 5, or 6; where: R 17A and R 18A is hydrogen or R 19A are each independently selected from R 19A is a hydrocarbyl (e.g., C 1-6 alkyl, alkenyl, alkynyl, or haloalkyl), carbocyclyl, and -(CH) s1 -heterocyclyl, and each R 19A is halogen, cyano, amino, hydroxy, C 1-6 Alkyl, and C1-6 Optionally and independently substituted with one or more of alkoxy; and s1 is 0, 1, 2, 3, 4, 5, or 6).

[0045] Preferably, the compound of formula (III) or a salt thereof is N(R 1A )(R 2A Alternatively, the compound of formula (IV) or a salt thereof is preferably such that Z is —N(R 17A )R 18A If Z is absent and Y is -N(R 17A )R 18A is bonded to the linker via Y.

[0046] In one embodiment, the NMT inhibitor is a compound of Formula (IIIa): [ka] (In the formula: n1 is 0 or 1; E 1 is C; W is a (1-4C)hydrocarbyl, aryl (e.g., phenyl), or heteroaryl group (e.g., pyridinyl); M is selected from C and N; R 3A , R 4A , and R 5A is hydrogen, R 12A , and R 12A independently selected from optionally substituted (1-3C)hydrocarbyl; R 12A is halogen, trifluoromethyl, cyano, thio, nitro, oxo, -OR 13A , -SR 13A , -C(O)R 13A , -C(O)OR 13A , -OC(O)R 13A , -NR 13A COR 14A , and R 15A are independently selected from; R13A and R 14A are each independently selected from hydrogen or (1-4C)hydrocarbyl (e.g., methyl); Ring D * is an optionally substituted nitrogen-containing 6- or 7-membered heterocycle, wherein ring D * Each substitutable carbon or nitrogen in 7A optionally and independently substituted by; R 7A are independently selected from hydrogen, (1-4C)hydrocarbyl, halogen, trifluoromethyl, cyano, thio, nitro, or oxo; R 8A is hydrogen; p1 is 0, 1, or 2, where R 4A The values ​​of may be the same or different; q1 is 3, where R 5A The values ​​of may be the same or different; and t1 is 0, 1, or 2, where R 7A The values ​​of may be the same or different).

[0047] Preferably, the compound of formula (IIIa) or a salt thereof is NR 8A The linker is attached via a group.

[0048] In one embodiment, the NMT inhibitor is (2,6-dichloro-4-(2-piperazin-1-yl-pyridin-4-yl)-N-(1,3,5-trimethyl-1H-pyraxol-4-yl)-benzenesulfonamide): [ka] or a salt thereof.

[0049] Preferably, (2,6-dichloro-4-(2-piperazin-1-yl-pyridin-4-yl)-N-(1,3,5-trimethyl-1H-pyraxol-4-yl)-benzenesulfonamide) or a salt thereof is bound to the linker via the NH group of the piperazinyl ring.

[0050] In one embodiment, the NMT inhibitor is 2,6-dichloro-N-(5-isobutyl-1,3-dimethyl-1H-pyrazol-4-yl(-4-(2-piperazin-1-yl-pyridin-4-yl)-benzenesulfonamide: [ka] or a salt thereof.

[0051] Preferably, 2,6-dichloro-N-(5-isobutyl-1,3-dimethyl-1H-pyrazol-4-yl)-4-(2-piperazin-1-yl-pyridin-4-yl)-benzenesulfonamide or a salt thereof is bonded to the linker via the NH group of the piperazinyl ring.

[0052] The compounds of formula (III), (IV), (IIIa), (2,6-dichloro-4-(2-piperazin-1-yl-pyridin-4-yl)-N-(1,3,5-trimethyl-1H-pyraxol-4-yl)-benzenesulfonamide), and 2,6-dichloro-N-(5-isobutyl-1,3-dimethyl-1H-pyrazol-4-yl)-4-(2-piperazin-1-yl-pyridin-4-yl)-benzenesulfonamide, are disclosed in WO 2010 / 026365, the entire contents of which are incorporated by reference for purposes of describing the synthesis and activity of NMT inhibitors.

[0053] In one embodiment, the NMT inhibitor is a compound of formula (V): [ka] (In the formula: n1 is 1 or 2; n2 is 1 or 2; X 1 is CR x and N; If present, R xis optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of hydrogen, halogen, —OH, —OCH3, and —OCF3; 1-4 selected from the group consisting of alkyl; R 1 is hydrogen; -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -OCH3, and -OCF3; 1-4 alkyl; and -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -CH3, -OCH3, and -OCF3. 3-6 cycloalkyl; R 2 is hydrogen; -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -OCH3, and -OCF3; 1-4 alkyl; and -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -CH3, -OCH3, and -OCF3. 3-6 is selected from the group consisting of cycloalkyl; or R 1 and R 2 together with the atoms to which they are bonded, they are C 3-6 are linked to form a cycloalkyl group or a 3- to 6-membered non-aromatic heterocyclyl group containing one heteroatom selected from the group consisting of O and N, wherein the C 3-6 The cycloalkyl group or 3- to 6-membered non-aromatic heterocyclyl group is optionally substituted with one or two substituents, each substituent independently selected from the group consisting of halogen, —OH, —CH3, —OCH3, and —OCF3; R 3 is hydrogen; -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -OH, -OCH3, and -OCF3; 1-4alkyl; and -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -OH, -CH3, -OCH3, and -OCF3. 3-6- is selected from the group consisting of cycloalkyl; or R 1 and R 3 are linked together with the atoms to which they are attached such that they form a 3- to 6-membered non-aromatic heterocyclyl group containing one N heteroatom, wherein the 3- to 6-membered non-aromatic heterocyclyl group is optionally substituted with one or two substituents, each substituent independently selected from the group consisting of halogen, —CH3, —OH, —OCH3, and —OCF3; X 2 is CR 4 and N; If present, R 4 is hydrogen; halogen; each of the substituents is halogen, -OH, -OCH3, -OCF3, and -NR a R b -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 selected from the group consisting of alkyl; R 5a and R 5d are independently selected from the group consisting of hydrogen; halogen; methyl optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -OH, -OCH3, and -OCF3; and methoxy optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -OH, -OCH3, and -OCF3; R 5b and R 5c is hydrogen; halogen; —C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, —OH, —OCH3, and —OCF3. 1-4alkyl; -OC optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -OH, -OCH3, and -OCF3; 1-4 alkyl; and C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, —CH3, —OH, —OCH3, and —OCF3. 3-6 independently selected from the group consisting of cycloalkyl; or R 5b and R 5c are linked together with the atoms to which they are attached such that they form a 6-membered aryl group or a 5- or 6-membered aromatic heterocyclyl group containing 1 or 2 heteroatoms selected from the group consisting of S, O, and N, wherein the 6-membered aryl group or the 5- or 6-membered aromatic heterocyclyl group is optionally substituted with 1 or 2 substituents, each substituent independently selected from the group consisting of halogen, —OH, —OCH3, and —OCF3; R 6 is selected from the group consisting of hydrogen and methyl; If present, each R 7 is —C optionally substituted with one, two, or three substituents, each substituent being independently selected from the group consisting of halogen, —OH, —OCH3, and —OCF3; 1-4 is alkyl; R 8 is hydrogen; halogen; -OH; -CN; -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -OH, -CN, and methoxy optionally substituted with one, two, or three halogens. 1-4 alkyl; -C optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -CH3, -OH, -CN, and methoxy optionally substituted with one, two, or three halogens; 3-6cycloalkyl; -C optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -OH, -CN, and methoxy optionally substituted with one, two, or three halogens; 1-4 alkenyl; and -OC optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -OH, -CN, and methoxy optionally substituted with one, two, or three halogens. 1-4 selected from the group consisting of alkyl; R 9 is —C optionally substituted with hydrogen and one, two, or three substituents, each of which is independently selected from the group consisting of halogen, —OH, —OCH3, and —OCF3; 1-4 is selected from the group consisting of alkyl; or R 8 and R 9 together with the atoms to which they are attached, they form a six-membered aryl group, C 5-6 are linked to form a cycloalkyl group or a 5- or 6-membered aromatic heterocyclyl group containing 1 or 2 heteroatoms selected from N, O, and S, wherein the 6-membered aryl group, C 5-6 The cycloalkyl group or the 5- to 6-membered aromatic heterocyclyl group is optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen; -OH; -CN; and methoxy, each of which is optionally substituted with one, two, or three halogens. 1-4 alkyl; and -OC optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of -halogen, -OH, and methoxy optionally substituted with one, two, or three halogens. 1-4 optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of alkyl; p is 0, 1, or 2; Z is a 5- to 13-membered non-aromatic heterocyclyl group containing 1, 2, or 3 heteroatoms selected from N, O, and S, where at least one of the heteroatoms is N, and where the 5- to 13-membered non-aromatic heterocyclyl group is selected from the group consisting of: each substituent being a halogen; each substituent being a halogen, —OH, and —OC optionally substituted with 1, 2, or 3 halogens. 1-3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 Alkyl; each substituent is halogen, -OH, and -OC optionally substituted with one, two, or three halogens. 1-3 -OC optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 Alkyl;NR c R d and -OC, each of which is optionally substituted with halogen, -OH, and one, two, or three halogens. 1-3 C optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 3-6 cycloalkyl; or when two substituents are at adjacent ring positions, they, together with the atoms to which they are attached, form a ring. 3-6 They may be linked to form a cycloalkyl group or a 4- to 6-membered non-aromatic heterocyclyl group containing one heteroatom selected from the group consisting of O and N, wherein the C 3-6 The cycloalkyl group or the 4- to 6-membered non-aromatic heterocyclyl group is optionally substituted with one or two substituents, each of which is a halogen; each of which is a halogen, —OH, and —OC optionally substituted with one, two, or three halogens. 1-3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6alkyl; and —OC, each of which is optionally substituted with halogen, —OH, and one, two, or three halogens. 1-3 -OC optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 independently selected from the group consisting of alkyl; R c is hydrogen; R d is hydrogen; -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -OCH3, and -OCF3; 1-6 alkyl; and C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, —CH3, —OH, —OCH3, and —OCF3. 3-6 is selected from the group consisting of cycloalkyl; or Z is -NR 10 R 11 where: R 10 is hydrogen; and R 11 is a 5- to 10-membered non-aromatic heterocyclyl group containing 1, 2, or 3 heteroatoms selected from N, O, and S, where at least one of the heteroatoms is N, and where the 5- to 10-membered non-aromatic heterocyclyl group is selected from the group consisting of halogen; —OH; —OC, each of which is optionally substituted with halogen, —OH, and 1, 2, or 3 halogens. 1-3 -C optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of alkyl 1-6 alkyl; and —OC, each of the substituents being optionally substituted with halogen, —OH, and 1, 2, or 3 halogens. 1-3 -OC optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 optionally substituted with 1, 2, 3, or 4 substituents independently selected from the group consisting of alkyl; and If present, each R a and Rb is hydrogen and -C 1-4 alkyl).

[0054] Suitably, the compound of formula (V) is a nitrogen atom present in group Z, for example a nitrogen ring atom (when group Z is a 5- to 13-membered non-aromatic heterocyclyl group), an NR c R d group (where the group Z is a 5- to 13-membered non-aromatic heterocyclyl group), or —NR 10 R 11 is attached to the linker via a linker linker group Z. Those skilled in the art will understand that in order for the linker to be attached to a nitrogen atom present in group Z (e.g., in compounds of formula (V) and elsewhere herein), the nitrogen atom present in group Z must be attached to at least one hydrogen atom, i.e., the nitrogen atom cannot be part of a tertiary amino group. The N-H covalent bond in group Z is replaced with an N-linker covalent bond in the conjugate formed between the compound (e.g., group Z) and the linker.

[0055] In one embodiment, the NMT inhibitor is a compound of Formula (Vq): or a salt thereof [ka] (In the formula: R 3a is H or C optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -OCH3, and -OCF3; 1-4 is alkyl; R 8a is optionally substituted with halogen or one, two, or three substituents, each substituent being independently selected from the group consisting of halogen, -OCH3, and -OCF3; 1-4 is alkyl; ra is 0, 1, or 2; m a is 1 or 2; R 12a is hydrogen; and If present, each R 13a is optionally substituted with one, two, or three substituents, each substituent being independently selected from the group consisting of halogen, -OCH3, and -OCF3; 1-4 alkyl; and C optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -CH3, -OCH3, and -OCF3. 3-6 independently selected from the group consisting of cycloalkyl; or ra is 2 and two R 13a When groups are located at adjacent ring positions, the two R 13a are each a halogen, -OH, and -OC optionally substituted with 1, 2, or 3 halogens, together with the atoms to which they are attached; 1-3 -C optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of alkyl 1-6 C optionally substituted with one or two substituents independently selected from the group consisting of alkyl 3-6 linked to form a cycloalkyl group).

[0056] Preferably, the compound of formula (Vq) or a salt thereof is NR 12a The linker is attached via a group.

[0057] In one embodiment, the NMT inhibitor is (S)-1-(5-chloro-2-(2-methylpiperazin-1-yl)pyrimidin-4-yl)-N-(2-(imidazo[1,2-a]pyridin-3-yl)propan-2-yl)azetidine-3-carboxamide (referred to herein as “NMT inhibitor 26”): [ka] or a salt thereof.

[0058] Suitably, (S)-1-(5-chloro-2-(2-methylpiperazin-1-yl)pyrimidin-4-yl)-N-(2-(imidazo[1,2-a]pyridin-3-yl)propan-2-yl)azetidine-3-carboxamide is attached to the linker via the NH group of the piperazinyl ring.

[0059] Compounds of formula (V), (Vq), and (S)-1-(5-chloro-2-(2-methylpiperazin-1-yl)pyrimidin-4-yl)-N-(2-(imidazo[1,2-a]pyridin-3-yl)propan-2-yl)azetidine-3-carboxamide are disclosed in WO 2022 / 058745, the entire contents of which are incorporated by reference for purposes of describing the synthesis and activity of NMT inhibitors. In particular, NMT inhibitor 26 can be prepared as described in Example 129 of WO 2022 / 058745.

[0060] In one embodiment, the NMT inhibitor is a compound of formula (VI): [ka] (In the formula: R 1 is a group of formula OLA; L is -(CHR 12 ) m - and; Each R 12 are independently H or C 1-4 is alkyl; m is 1, 2, or 3; A is, [ka] : and v is 0, 1, or 2; R 9a is H, C 1-4 Alkyl or C 1-4 is haloalkyl; R 9b is H, C 1-4 Alkyl or C 1-4 is haloalkyl; R 9c is C 1-4 Alkyl or C 1-4 is haloalkyl; R 9d is H, C 1-4 Alkyl or C 1-4 is haloalkyl; R 10 is H, C 1-4 Alkyl or C 1-4 is haloalkyl; R 11 H, halo, CN, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy or C 1-4 haloalkoxy; s is 0, 1, 2, or 3; Each R 2 are independently F, Cl, Br, C optionally substituted with up to three halogen groups 1-4 alkyl, OCH3, or OCF3; Y is CH or C 1-4 is alkyl; R 3 is H or C 1-4 is alkyl; R 4 is H or C 1-4 is alkyl; R 5 is H; R 6 is H or C 1-4 is alkyl; q is 0 or 1; R 7 is H or methyl; R 8 is H or methyl; or R 3 and R 6 and an intervening atom, the intervening atom and a bond, or the intervening atom and -(CHR a ) r -, or R 7 Groups and R 6The group and the intervening atom are a ) r -, forming a 3- to 7-membered non-aromatic heterocycle; r is 1, 2, 3, 4, or 5; and R a is hydrogen or methyl).

[0061] Suitably, the compound of formula (VI) or a salt thereof is N(R 5 )(R 6 ) group to the linker.

[0062] In one embodiment, the NMT inhibitor is a compound of formula (VIA): [ka] (In the formula: R 2a is H or F; R 2b is F; R 5a is H; R 6a is H or methyl; R 9ca is methyl, iso-propyl, or tert-butyl; R 9cb is H or methyl; R 10a is methyl; and R 11a is methyl; However, R 2a When is H, R 9cb is H).

[0063] Suitably, the compound of formula (VIA) or a salt thereof is N(R 5a )(R 6a ) group to the linker.

[0064] In one embodiment, the NMT inhibitor is 1-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}-2,2-dimethylpropan-1-ol: [ka] or a salt thereof.

[0065] Preferably, 1-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}-2,2-dimethylpropan-1-ol or a salt thereof is bound to the linker via the NH(Me) group.

[0066] In one embodiment, the NMT inhibitor is 2-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}propan-2-ol: [ka] or a salt thereof.

[0067] Preferably, 2-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}propan-2-ol or a salt thereof is bound to the linker via the NH(Me) group.

[0068] The compounds of formula (VI), (VIA), 1-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}-2,2-dimethylpropan-1-ol, and 2-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}propan-2-ol, are disclosed in EP Patent Application No. 22194959.7, the entire contents of which are incorporated by reference for purposes of describing the synthesis and activity of NMT inhibitors. The compounds may also be prepared according to methods known to those skilled in the art, for example, the methods disclosed herein for the synthesis of NMT inhibitor 1.

[0069] In one embodiment of the present invention, the NMT inhibitor is provided in the form of a salt. In one embodiment, the NMT inhibitor is provided in the form of a pharmaceutically acceptable salt. In one embodiment, an NMT inhibitor is provided.

[0070] (Linker) The linker connects the antibody to the NMT inhibitor via a covalent bond. The linker is a bifunctional or multifunctional moiety that can be used to link one or more NMT inhibitors with the antibody to form the ADC of the present invention. The linker can be stable outside the cell, i.e., extracellularly, or it can be cleavable by enzymatic activity, hydrolysis, or other metabolic conditions. In one embodiment, the linker is a cleavable linker, for example, cleavable by an enzyme (particularly a lysosomal enzyme).

[0071] The ADCs of the invention can be conveniently prepared using a linker having a reactive functional group for coupling to an NMT inhibitor and an antibody: A cysteine ​​thiol or amine, e.g., a chain terminus such as the N-terminus, or an amino acid side chain such as lysine, of the antibody can form a bond with a functional group, e.g., maleimide, of the linker.

[0072] The linker is preferably stable outside the cell. Prior to transport or delivery into the cell, the ADC of the present invention preferably remains stable and intact, i.e., the antibody remains linked to the NMT inhibitor. The linker is stable outside the target cell and can be cleaved at an effective rate inside the cell. An effective linker: (i) maintains the specific binding properties of the antibody; (ii) enables intracellular delivery of the conjugate or drug moiety; (iii) remains stable and intact, i.e., is not cleaved, until the conjugate has been delivered or transported to its target site; and (iv) once cleaved, maintains the cytotoxic cell-killing or cytostatic effect of the NMT inhibitor. The stability of the ADC can be measured by standard analytical techniques, such as mass spectrometry, HPLC, and LC / MS separation / analysis techniques. From derivatization studies (data not shown), applicants have concluded that the ADCs of the present invention do not have NMT inhibitory activity and therefore NMT inhibitory activity is realized upon cleavage of the linker, releasing the NMT inhibitor.

[0073] Covalent attachment of an antibody to a drug moiety requires that the linker have two reactive functional groups, i.e., bivalency in the sense of reactivity. Bivalent linker reagents useful for attaching two or more functional or biologically active moieties, such as peptides, nucleic acids, drugs, toxins, antibodies, haptens, and reporter groups, are known, and methods for obtaining the resulting conjugates have been reported (Hermanson, GT (1996) Bioconjugate Techniques; Academic Press: New York, pp. 234-242).

[0074] The linker may be substituted with groups that modify aggregation, solubility, or reactivity. For example, sulfonate substituents can increase the water solubility of the reagent and, depending on the synthetic route utilized to prepare the ADC, facilitate the coupling reaction between the linker reagent and an antibody or drug moiety, or between the linker and NMT inhibitor and the antibody or linker, and between the antibody and the NMT inhibitor.

[0075] In one embodiment, the linker unit has formula (VII): -A a -W w -Y y -(Formula (VII)) (In the formula: A, if present, is a First Stretcher unit that forms a covalent bond with a chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of an antibody; a is 0 or 1; each W is independently an amino acid unit or a glucuronide unit that, when A and / or Y are absent, forms a covalent bond with a chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of the antibody and / or with a functional group of an NMT inhibitor, respectively; When W is an amino acid unit, w is 1 to 12; When W is a glucuronide unit, w is 1 or 2; Y, when present, is a second Stretcher unit that forms a covalent bond with a functional group of the NMT inhibitor; and y is 0 or 1) It has.

[0076] The NMT inhibitor forms a covalent bond with the second Stretcher unit (Y), if present, or with the amino acid unit or glucuronide unit (W), if Y is absent. Preferably, the functional group on the NMT inhibitor is amino or alcohol, e.g., amino. When the NMT inhibitor forms a covalent bond via an amino group, the amino group must have an available hydrogen atom to allow reaction with the corresponding functional group (e.g., carbonyl group) in the linker; i.e., the amino group cannot be tertiary.

[0077] (First Stretcher Unit) The Stretcher unit (A), when present, can link the antibody to the amino acid unit or glucuronide unit (W). In this regard, the antibody has a functional group that can form a bond with a functional group of the Stretcher unit, e.g., a functional group of an amino side chain of the antibody. Useful functional groups that can be present on the antibody naturally or through chemical manipulation include, but are not limited to, sulfhydryl (-SH), amino, hydroxyl, carboxy, anomeric hydroxyl groups of carbohydrates, and carboxyl. In some embodiments, the antibody functional group is sulfhydryl and / or amino, particularly sulfhydryl. Sulfhydryl groups can be generated by reduction of intramolecular disulfide bonds of the antibody. Sulfhydryl groups can also be generated by reaction of amino groups of lysine moieties of the antibody with 2-iminothiolane (Traut's reagent) or another sulfhydryl-generating reagent.

[0078] In one embodiment, the first Stretcher unit forms a bond with a sulfur atom of the antibody, which can be derived from a sulfhydryl group of the antibody.

[0079] In one embodiment, the stretcher unit (A) has the formula (A1): [ka] (In the formula: n is 1 to 6; [ka] represents the chain terminus (e.g., N-terminus) or point of attachment to a functional group of an amino acid side chain of an antibody; and [ka] represents the point of attachment to W) It has.

[0080] In one embodiment, A is (A2): [ka] is.

[0081] In one embodiment, A is (A3): [ka] is.

[0082] (amino acid and glucuronide units) Each W is independently an amino acid unit or a glucuronide unit that, when A and / or Y are absent, forms a covalent bond with the chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of the antibody and / or with a functional group of the NMT inhibitor, respectively.

[0083] In some embodiments, each W is independently an amino acid unit. In such embodiments, w is 1 to 12, e.g., 1, 2, or 3, particularly 2.

[0084] In some embodiments, the amino acid unit can be cleaved by one or more enzymes, e.g., tumor-associated proteases, thereby facilitating the release of the NMT inhibitor from the ADC upon exposure to intracellular proteases, e.g., lysosomal enzymes (Doronina et al. (2003) Nat. Biotechnol. 21:778-784).

[0085] In one embodiment, each W has the formula (WI): [ka] wherein w is as defined above; and R 19is H, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, CH2OH, CH(OH)Me, CH2CH2SMe, CH2C(O)SMe, CH2C(O)NH2, CH2C(O)OH, CH2CH2C(O)NH2, CH2CH2C(O)OH, (CH2)3NHC(=NH)NH2, (CH2)3NH2, (CH2)3NHC(O)Me, (CH2)3NHCHO, (CH2)4NHC(=NH)NH2, (CH2)4NH2, (CH2)4NHC(O)Me, (CH2)4NHCHO, (CH2)3NHC(O)NH2, (CH2)4NHC(O)NH2, CH2CH2CH(OH)CH2NH2, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, phenyl, [ka] and; where: [ka] represents the point of attachment to A (if present) or to the chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of the antibody; and [ka] represents the point of attachment to Y (if present) or to a functional group on the NMT inhibitor It has.

[0086] In one embodiment, at least one R 19 is iso-propyl. In one embodiment, at least one R 19 is (CH2)3NHC(=NH)NH2.

[0087] In one embodiment, (W)w is a group represented by formula (WII): [ka] (In the formula: R 19ais H, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, CH2OH, CH(OH)Me, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, phenyl, [ka] and R 19b is CH2CH2SMe, CH2C(O)SMe, CH2C(O)NH2, CH2C(O)OH, CH2CH2C(O)NH2, CH2CH2C(O)OH, (CH2)3NHC(=NH)NH2, (CH2)3NH2, (CH2)3NHC(O)Me, (CH2) 3NHCHO, (CH2)4NHC(=NH)NH2, (CH2)4NH2, (CH2)4NHC(O)Me, (CH2)4NHCHO, (CH2)3NHC(O)NH2, (CH2)4NHC(O)NH2, or CH2CH2CH(OH)CH2NH2) It has.

[0088] In one embodiment, R 19a is iso-propyl. In one embodiment, R 19b is (CH2)3NHC(=NH)NH2.

[0089] In one embodiment, each W is a glucuronide unit. In such an embodiment, w is 1 or 2, particularly 1. The glucuronide unit contains a site that can be cleaved by a β-glucuronidase enzyme.

[0090] In one embodiment, W (particularly (W)w) is of formula (WIII): [ka] (In the formula: R is H, halo, CN, or NO; m is 0, 1, 2, or 3; Su is a sugar moiety; where: [ka] represents the point of attachment to A (if present) or to the chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of the antibody; and [ka] represents the point of attachment to Y (if present) or to a functional group on the NMT inhibitor It has.

[0091] In one embodiment, R is H. In a second embodiment, R is halo. In a third embodiment, R is CN. In a fourth embodiment, R is NO.

[0092] In one embodiment, m is 0. In a second embodiment, m is 1. In a third embodiment, m is 2. In a fourth embodiment, m is 3.

[0093] In one embodiment, the sugar moiety (Su) is a cyclic hexose, e.g., a pyranose, or a cyclic pentose, e.g., a furanose. In one embodiment, the pyranose is a glucuronide or a hexose. Suitably, the sugar moiety is in the β-D form. In one embodiment, the pyranose is a β-D-glucuronide moiety (i.e., a β-D-glucuronic acid linked to the remainder of W via a glycosidic bond that is cleavable by β-glucuronidase). In one embodiment, the sugar moiety is unsubstituted (e.g., a naturally occurring cyclic hexose or cyclic pentose). In one embodiment, the sugar moiety is a substituted β-D-glucuronide (e.g., a cyclic hexose or cyclic pentose containing one or more groups such as hydrogen, hydroxyl, halogen, sulfur, nitrogen, or C). 1-6 alkyl-substituted glucuronic acid).

[0094] In one embodiment, Su has the formula: [ka] It has.

[0095] (Second Stretcher Unit) The second stretcher unit (Y), when present, links W to the NMT inhibitor by forming a covalent bond with a functional group of the NMT inhibitor. For example, Y may contain a carbonyl group that can form a covalent bond with an amino functional group of the NMT inhibitor. When the NMT inhibitor forms a covalent bond through an amino group, the amino group must have an available hydrogen atom to allow reaction with the corresponding functional group (e.g., carbonyl group) of Y; i.e., the amino group cannot be tertiary. The second stretcher unit may be a "self-immolative" or "non-self-immolative" group.

[0096] A "non-self-immolative" Stretcher unit is one in which some or all of the Stretcher unit remains attached to the drug moiety upon enzymatic (e.g., proteolytic) cleavage of the ADC. Examples of non-self-immolative spacer units include, but are not limited to, a glycine stretcher unit and a glycine-glycine stretcher unit. Other combinations of peptidic Stretcher units that are susceptible to sequence-specific enzymatic cleavage are also contemplated. For example, enzymatic cleavage of an ADC containing a glycine-glycine stretcher unit by a tumor cell-associated protease results in the release of the glycine-glycine-NMT inhibitor moiety from the remainder of the ADC. In one such embodiment, the glycine-glycine-NMT inhibitor moiety subsequently undergoes a separate hydrolysis step within the tumor cell, resulting in cleavage of the glycine-glycine spacer unit from the NMT inhibitor moiety.

[0097] The "self-immolative" spacer unit allows for the release of the NMT inhibitor without a separate hydrolysis step.

[0098] When W is an amino acid unit, preferably y is 1. When W is a glucuronide unit, preferably y is 0.

[0099] In one embodiment, the second stretcher unit (Y) has the formula (Y1): [ka] (In the formula: Each Q is independently halo, NO2, CN, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 haloalkoxy; m is 0 to 4; where: [ka] represents the point of attachment to W; and [ka] represents the point of attachment to the functional group of the NMT inhibitor) It has.

[0100] In one embodiment, m is not 0 and at least one Q is halo. In a second embodiment, m is not 0 and at least one Q is NO. In a third embodiment, m is not 0 and at least one Q is CN. In a fourth embodiment, m is not 0 and at least one Q is C. 1-6 In a fifth embodiment, m is not 0 and at least one Q is C 1-6 In a sixth embodiment, m is not 0 and at least one Q is C 1-6 In a seventh embodiment, m is not 0 and at least one Q is C 1-6 It is haloalkoxy.

[0101] In one embodiment, m is 0. In a second embodiment, m is 1. In a third embodiment, m is 2. In a fourth embodiment, m is 3. In a fifth embodiment, m is 4.

[0102] Suitable examples of self-immolative Stretcher units further include, but are not limited to, p-aminobenzyl alcohol and its derivatives, such as formula (Y1) (see, e.g., US 2005 / 0256030 A1), such as 2-aminoimidazole-5-methanol derivatives (Hay et al. (1999) Bioorg. Med. Chem. Lett. 9:2237), and aromatic compounds electronically similar to ortho- or para-aminobenzyl acetals. Spacers that undergo cyclization upon amide bond hydrolysis, such as substituted and unsubstituted 4-aminobutyric acid amides (Rodrigues et al., Chemistry Biology, 1995, 2, 223); appropriately substituted bicyclo[2.2.1] and bicyclo[2.2.2] ring systems (Storm et al., J. Amer. Chem. Soc., 1972, 94, 5815); and 2-aminophenylpropionic acid amide (Amsbury et al., J. Org. Chem., 1990, 55, 5867). Elimination of amine-containing drugs substituted at the a-position of glycine (Kingsbury et al., J. Med. Chem., 1984, 27, 1447) is also an example of a self-immolative stretcher unit useful in ADCs.

[0103] In one embodiment, the linker has the formula (LI): [ka] (In the formula, [ka] represents the point of attachment to the chain terminus (e.g., N-terminus) or functional group on an amino acid side chain of the antibody; and [ka] represents the point of attachment to the functional group of the NMT inhibitor) It has.

[0104] In one embodiment, the linker has formula (LII): [ka] (In the formula, [ka] represents the point of attachment to the chain terminus (e.g., N-terminus) or functional group on an amino acid side chain of the antibody; and [ka] represents the point of attachment to the functional group of the NMT inhibitor) It has.

[0105] In one embodiment, the linker has formula (LIII): [ka] (In the formula, [ka] represents the point of attachment to the chain terminus (e.g., N-terminus) or functional group on an amino acid side chain of the antibody; and [ka] represents the point of attachment to the functional group of the NMT inhibitor) It has.

[0106] In one embodiment, the linker has the formula (LIV): [ka] (In the formula, [ka] represents the point of attachment to the chain terminus (e.g., N-terminus) or functional group on an amino acid side chain of the antibody; and [ka] represents the point of attachment to the functional group of the NMT inhibitor) It has.

[0107] (antibody) The term "antibody" as used herein refers to monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), intact antibodies, and antibody fragments, so long as they exhibit the desired biological activity. Antibodies may be murine, human, humanized, chimeric, or derived from other species. Antibodies are proteins produced by the immune system that can recognize and bind to specific antigens (Janeway, C., Travers, P., Walport, M., Shlomchik (2001) Immuno Biology, 5th ed., Garland Publishing, New York). A target antigen usually has multiple binding sites, also called epitopes, recognized by CDRs on multiple antibodies. Each antibody that specifically binds to a different epitope has a different structure. Therefore, one antigen can have multiple corresponding antibodies. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds to an antigen or portion thereof of a target of interest, including, but not limited to, cancer cells or cells that produce autoimmune antibodies associated with autoimmune disease. The immunoglobulin can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) or subclass, or allotype (e.g., human G1m1, G1m2, G1m3, non-G1m1 [i.e., any allotype other than G1m1], G1m17, G2m23, G3m21, G3m28, G3m11, G3m5, G3m13, G3m14, G3m10, G3m15, G3m16, G3m6, G3m24, G3m26, G3m27, A2m1, A2m2, Km1, Km2, and Km3) of immunoglobulin molecule. The immunoglobulin can be from any species, including human, murine, or rabbit origin.

[0108] "Antibody fragments" comprise portions of a full-length antibody, typically the antigen-binding or variable region thereof. Examples of antibody fragments include Fab, Fab', F(ab'), and scFv fragments; diabodies; linear antibodies; fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies, CDRs (complementarity-determining regions), and epitope-binding fragments of any of the above that immunospecifically bind to cancer cell, viral, or microbial antigens, single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0109] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible natural mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations which include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous in that they may be synthesized uncontaminated by other antibodies. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies used in accordance with the present invention may be produced by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, or by recombinant DNA methods (see U.S. Pat. No. 4,816,567). Monoclonal antibodies may also be isolated from phage antibody libraries using the techniques described in Clackson et al. (1991) Nature 352:624-628; Marks et al. (1991) J. Mol. Biol. 222:581-597, or from transgenic mice carrying a fully human immunoglobulin system (Lonberg (2008) Curr. Opinion 20(4):450-459).

[0110] The monoclonal antibodies herein specifically include "chimeric" antibodies in which portions of the heavy and / or light chains are identical to or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of these chains are identical to or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al. (1984) Proc. Natl. Acad. Sci. USA, 81:6851-6855). Chimeric antibodies include "primatized" antibodies containing variable domain antigen-binding sequences derived from a non-human primate (e.g., an Old World monkey or ape) and human constant region sequences.

[0111] An "intact antibody" herein comprises a VL domain and a VH domain, as well as a light chain constant domain (CL) and heavy chain constant domains CH1, CH2, and CH3. The constant domains may be native-sequence constant domains (e.g., human native-sequence constant domains) or amino acid sequence variants thereof. An intact antibody may have one or more "effector functions," which refer to biological activities attributable to the Fc region (a native-sequence Fc region or amino acid sequence variant Fc region) of an antibody. Examples of antibody effector functions include C1q binding; complement-dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; and down-regulation of cell surface receptors such as B cell receptors and BCRs.

[0112] Depending on the amino acid sequence of the constant domain of their heavy chain, intact antibodies can be assigned to different "classes." There are five major classes of intact antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into "subclasses" (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The heavy-chain constant domains that correspond to the different classes of antibodies are called α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[0113] The desired biological activity of the antibody preferably includes binding to a cancer-associated antigen, which binding is preferably specific, i.e., the antibody binds preferentially to the cancer-associated antigen and does not significantly bind to other antigens or proteins (e.g., BSA).

[0114] As used herein, a cancer-associated antigen is an antigen expressed by a cancer cell. The cancer cell may be a tumor cell or a cell of a blood cancer, such as a B cell or a T cell. Thus, an antibody targeting an antigen expressed by a cancer cell binds to the cancer cell and (without being limited by theory) mediates cell death through the action of an NMT inhibitor and / or antibody-mediated cell-mediated cytotoxicity (ADCC). Although cancer-associated antigens can also be expressed by non-cancerous cells, they are preferably preferentially expressed by cancerous cells. An example of a cancer-associated antigen is HER2. HER2 is particularly expressed on some breast cancer cells. Another example of a cancer-associated antigen is CD20. CD20 is expressed on some B cells, including some cells of B-cell lymphomas.

[0115] As used herein, "binds to HER2" means that the antibody binds to HER2 with higher affinity than a non-specific partner, such as bovine serum albumin (BSA, GenBank Accession No. CAA76847, Version No. CAA76847.1 GI:3336842, Archive Last Updated: January 7, 2011, 2:30 PM). In some embodiments, the antibody binds to HER2 with an association constant (Ka) that is at least 2, 3, 4, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, 10, 10, or 10 times higher than the association constant of the antibody for BSA, when measured under physiological conditions. The antibodies of the present invention can bind to HER2 with high affinity. For example, in ...0, 1000, 2000, 5000, 10, 10, or 10 times higher than the association constant of the antibody for BSA. -6 M or less, e.g., 1×10 -6 , 10 -7 , 10 -8 , 10 -9 ,10 -10 , 10-11 , 10 -12 , 10 -13 , or 10 -14 It can bind to HER2 with a KD of

[0116] As used herein, HER2 refers to human epidermal growth factor receptor 2. In one embodiment, the HER2 polypeptide corresponds to GenBank Accession No. AAA75493, Version No. AAA75493.1 GI:306840, Record Last Modified: June 23, 2010, 8:47 AM. In one embodiment, the nucleic acid encoding the HER2 polypeptide corresponds to GenBank Accession No. M11730, Version No. Version No. M11730.1 GL183986, Record Last Modified: June 23, 2010, 8:47 AM.

[0117] (Antibody Modification) The antibodies disclosed herein can be modified, for example, to make them less immunogenic to human subjects. This can be accomplished using any of a number of techniques well known to those of skill in the art. Some of these techniques are described in more detail below.

[0118] (humanized) Techniques for reducing the in vivo immunogenicity of non-human antibodies or antibody fragments include those referred to as "humanization." A "humanized antibody" refers to a polypeptide comprising at least a portion of a modified variable region of a human antibody, in which a portion of the variable region, preferably substantially less than an intact human variable domain, has been replaced by the corresponding sequence from a non-human species, and the modified variable region is linked to at least another portion of another protein, preferably a constant region of a human antibody. The term "humanized antibody" includes human antibodies in which one or more complementarity-determining region ("CDR") amino acid residues and / or one or more framework region ("FW" or "FR") amino acid residues have been replaced by amino acid residues from analogous sites in rodent or other non-human antibodies. The term "humanized antibody" also includes immunoglobulin amino acid sequence variants or fragments thereof comprising FRs having substantially the amino acid sequence of a human immunoglobulin and CDRs having substantially the amino acid sequence of a non-human immunoglobulin.

[0119] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. Or, viewed another way, a humanized antibody is a human antibody that also contains selected sequences of non-human (e.g., murine) antibodies in place of human sequences. A humanized antibody can include conservative amino acid substitutions or non-naturally occurring residues from the same or different species that do not significantly alter its binding and / or biological activity. Such antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin.

[0120] There are various humanization techniques, including "CDR grafting", "guide selection", "deimmunization", "resurfacing" (also known as "veneering"), "composite antibodies", "human string content optimization", and framework shuffling.

[0121] (CDR port) In this technique, humanized antibodies are human immunoglobulins (recipient antibodies) in which residues from the complementarity-determining regions (CDRs) of the recipient antibody are replaced by residues from a CDR (donor antibody) of a non-human species such as mouse, rat, camel, cow, goat, or rabbit having the desired properties (in effect, the non-human CDRs are "grafted" onto a human framework). In some instances, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues (this can occur, for example, when particular FR residues have a significant effect on antigen binding).

[0122] Furthermore, humanized antibodies can comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications are made to further refine and maximize antibody performance. Thus, in general, a humanized antibody will comprise at least one, and in one aspect all two, variable domains, in which all or all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin sequence. The humanized antibody optionally also will comprise at least a portion of an immunoglobulin constant region (Fc), or at least a portion of a human immunoglobulin constant region (Fc).

[0123] (Guide Selection) This method involves combining the VH or VL domain of a given non-human antibody specific for a particular epitope with a human VH or VL library, and selecting specific human V domains against the antigen of interest. This selected human VH is then combined with a VL library to generate fully human VH x VL combinations. This method is described in Nature Biotechnology (NY) 12, (1994) 899-903.

[0124] (complex antibody) In this method, two or more segments of human antibody amino acid sequence are combined in the final antibody molecule. These are constructed by combining multiple human VH and VL sequence segments in combinations that limit or avoid human T cell epitopes in the V regions of the final composite antibody. Where necessary, T cell epitopes are limited or avoided by replacing V region segments that contribute to or encode T cell epitopes with alternative segments that avoid the T cell epitopes. This method is described in US 2008 / 0206239 A1.

[0125] (Deimmunization) This method involves removing human (or other second species) T cell epitopes from the V region of a therapeutic antibody (or other molecule). The V region sequence of the therapeutic antibody is analyzed for the presence of MHC class II binding motifs, for example, by comparison with a database of MHC binding motifs (e.g., the "Motif" database hosted at www.wehi.edu.au). Alternatively, MHC class II binding motifs can be identified using computer-threaded methods such as those devised by Altuvia et al. (J. Mol. Biol. 249 244-250 (1995)); these methods test consecutive overlapping peptides derived from the V region sequence for their binding energy to MHC class II proteins. This data can then be combined with information on other sequence features associated with successfully presented peptides, such as amphipathicity, Rothbard motifs, and cleavage sites for cathepsin B and other processing enzymes.

[0126] Once a potential second species (e.g., human) T-cell epitope has been identified, it is eliminated by modifying one or more amino acids. The modified amino acids are usually within the T-cell epitope itself, but may also be adjacent to the epitope in terms of the primary or secondary structure of the protein (and therefore may not be adjacent in the primary structure). Most typically, the modification is by substitution, although in some circumstances, the addition or deletion of amino acids may be more appropriate.

[0127] All modifications can be achieved by recombinant DNA technology, so that the final molecule can be prepared by expression from a recombinant host using well-established methods such as site-directed mutagenesis, although the use of protein chemistry or any other means of molecular modification is also possible.

[0128] (Resurfacing) The method includes: (a) determining the three-dimensional structure of a variable region (or fragment thereof) of a non-human (e.g., rodent) antibody by constructing a three-dimensional model of the variable region of the non-human antibody; (b) generating a sequence alignment using relative accessibility distributions obtained from x-ray crystal structures of heavy and light chains of a sufficient number of non-human and human antibody variable regions to obtain a set of heavy and light chain framework positions whose aligned positions are identical in 98% of the non-human antibody heavy and light chains; (c) defining a set of surface-exposed amino acid residues in the heavy and light chains for the non-human antibody to be humanized using the set of framework positions generated in step (b); (d) identifying from a human antibody amino acid sequence a set of surface-exposed amino acid residues of the heavy and light chains that are most closely identical to the set of surface-exposed amino acid residues defined in step (c), wherein the heavy and light chains from the human antibody are naturally paired or not naturally paired; (e) replacing the set of surface-exposed amino acid residues of the heavy and light chains defined in step (c) with the set of surface-exposed amino acid residues of the heavy and light chains identified in step (d) in the amino acid sequence of the non-human antibody to be humanized; (f) constructing a three-dimensional model of the variable region of the non-human antibody resulting from the substitutions specified in step (e); (g) comparing the three-dimensional models constructed in steps (a) and (f), identifying any amino acid residue from the set identified in step (c) or (d) that is within 5 angstroms of any atom of any residue in the complementarity determining region of the non-human antibody to be humanized; and (h) changing any residues identified in step (g) from a human amino acid residue to the original non-human amino acid residue, thereby defining a non-human antibody humanized set of surface-exposed amino acid residues; provided that step (a) does not have to be performed first, but must be performed before step (g).

[0129] (Super Humanized) This method compares non-human sequences with functional human germline gene repertoires. Human genes encoding the same or closely related canonical structures as the non-human sequences are selected. The selected human genes with the highest homology within the CDRs are selected as FR donors. Finally, non-human CDRs are grafted onto these human FRs. This method is described in WO 2005 / 079479 A2.

[0130] (Human string content optimization) This method compares a non-human (e.g., mouse) sequence with a repertoire of human germline genes and scores the differences as a human string content (HSC), which quantifies the sequence at the level of potential MHC / T cell epitopes. The target sequence is then humanized by maximizing its HSC, rather than using a global identity measure, to generate a large number of diverse humanized variants (described in Molecular Immunology, 44, (2007) 1986-1998).

[0131] (Framework Shuffling) The CDRs of a non-human antibody are fused in-frame to a cDNA pool encompassing all known human heavy and light chain germline gene frameworks. Humanized antibodies are then selected, for example, by panning a phage-display antibody library. This is described in Methods 36, 43-60 (2005).

[0132] In one embodiment, known antibodies for the treatment or prevention of cancer are used in accordance with the present invention. Examples of antibodies that can be used to treat cancer include, but are not limited to, trastuzumab, a humanized anti-HER2 monoclonal antibody for the treatment of patients with metastatic breast cancer (Stebbing, J., Copson, E., and O'Reilly, S., "Herceptin (trastuzumab) in advanced breast cancer," Cancer Treat Rev (2000). 26, 287-90; Miller et al. (2003) Journal of Immunology 170, 4854-4861), and rituximab, a chimeric anti-CD20 monoclonal antibody for the treatment of patients with non-Hodgkin's lymphoma.Further examples include oregovomab (OvaRex, AltaRex Corporation, MA), a murine antibody for the treatment of ovarian cancer; Panorex (Glaxo Wellcome, NC), a murine IgG2a antibody for the treatment of colorectal cancer; BEC2 (hnClone Systems, NY), a murine IgG antibody for the treatment of lung cancer; IMC-C225 (Imclone Systems, NY), a chimeric IgG antibody for the treatment of head and neck cancer; Vitaxin (MethnMune, MD), a humanized antibody for the treatment of sarcoma; Campas1 / H (Leukosite, MA), a humanized IgGi antibody for the treatment of chronic lymphocytic leukemia (CLL); Smart MI95 (Protein Design Labs, CA), a humanized IgG antibody for the treatment of acute myeloid leukemia (AML); LymphoCide (Immunomedics, NJ), a humanized IgG antibody for the treatment of non-Hodgkin's lymphoma; Smart ID 10 (Protein Design Labs, CA), a humanized antibody for the treatment of non-Hodgkin's lymphoma. Design Labs, CA); Oncolym (Techniclone, CA), a murine antibody for the treatment of non-Hodgkin's lymphoma; Allomune (BioTransplant, CA), a humanized anti-CD2 mAh for the treatment of Hodgkin's disease or non-Hodgkin's lymphoma; anti-VEGF (Genentech, CA), a humanized antibody for the treatment of lung and colorectal cancer; CEAcide (Immunomedics, NJ), a humanized anti-CEA antibody for the treatment of colorectal cancer; IMC-1C11 (ImClone Systems, NJ), an anti-KDR chimeric antibody for the treatment of colorectal cancer, lung cancer, and melanoma; and cetuximab (ImClone, NJ), an anti-EGFR chimeric antibody for the treatment of epidermal growth factor-positive cancers. Further examples of antibodies that bind to CD20 include ocrelizumab, obinutuzumab, ofatumumab, ibritumomab tiuxetan, tositumomab, and ublituximab.

[0133] Other antibodies useful in treating cancer include antibodies against the following cancer-associated antigens: CA125 (ovarian), CA15-3 (epithelial cancer), CAI9-9 (epithelial cancer), L6 (epithelial cancer), Lewis Y (epithelial cancer), Lewis X (epithelial cancer), alpha-fetoprotein (epithelial cancer), CA 242 (colorectal), placental alkaline phosphatase (epithelial cancer), prostate-specific antigen (prostate), prostatic acid phosphatase (prostate), epidermal growth factor (epithelial cancer), MAGE-1 (epithelial cancer), MAGE-2 (epithelial cancer), MAGE-3 (epithelial cancer), MAGE-4 (epithelial cancer), anti-transferrin receptor (epithelial cancer), p97 (melanoma), MUC1-KLH (breast cancer), CEA (colorectal), gplOO (melanoma), MARTI (melanoma), and PSA. (prostate), IL-2 receptor (T-cell leukemia and lymphoma), CD19 (B-cell lymphoma), CD20 (non-Hodgkin's lymphoma), CD52 (leukemia), CD33 (leukemia), CD22 (lymphoma), human chorionic gonadotropin (epithelial cancer), CD38 (multiple myeloma), CD40 (lymphoma), mucin (epithelial cancer), P21 (epithelial cancer), MPG (melanoma), and Neu oncogene product (epithelial cancer). Some specific and useful antibodies include BR96 mAb (Trail, PA, Willner, D., Lasch, SJ, Henderson, AJ, Hofstead, SJ, Casazza, AM, Firestone, RA, Hellstrom, I., Hellstrom, KE, "Cure of Xenografted Human Carcinomas by BR96-Doxorubicin Immunoconjugates," Science 1993, 261, 212-215), BR64 (Trail, PA, Willner, D, Knipe, J., Henderson, AJ, Lasch, SJ, Zoeckler, ME, Trailsmith, MD, Doyle, TW, King, HD, Casazza, AM, Braslawsky, GR, Brown, JP), and others., Hofstead, SJ, Greenfield, RS, Firestone, RA, Mosure, K., Kadow, DF, Yang, MB, Hellstrom, KE, and Hellstrom, I., "Effect of Linker Variation on the Stability, Potency, and Efficacy of Carcinoma-reactive BR64-Doxorubicin Immunoconjugates," Cancer Research 1997, 57, 100-105; mAbs against the CD40 antigen, such as S2C6 mAb (Francisco, JA, Donaldson, KL, Chace, D., Siegall, CB, and Wahl, AF, "Agonistic properties and in vivo antitumor activity of the anti-CD40 antibody SGN-14"); Examples of internalizing antibodies that bind to tumor-associated antigens include, but are not limited to, mAbs against the CD70 antigen, such as the anti-CD40 antibody, SGN-14, Cancer Res. 2000, 60, 3225-3231), mAbs against the CD70 antigen, such as 1F6 mAb, and mAbs against the CD30 antigen, such as AGIO (Bowen, MA, Olsen, KJ, Cheng, L., Avila, D., and Podack, ER, "Functional effects of CD30 on a large granular lymphoma cell line YT," J. Immunol., 151, 5896-5906, 1993). Many other internalizing antibodies that bind to tumor-associated antigens can be used in the present invention and have been reviewed (Franke, AE, Sievers, EL, and Scheinberg, DA"Cell surface receptor-targeted therapy of acute myeloid leukemia: a review," Cancer Biother Radiopharm. 2000, 15, 459-76; Murray, JL, "Monoclonal antibody treatment of solid tumors: a coming of age," Semin Oncol. 2000, 27, 64-70; Breitling, F. and Dubel, S., Recombinant Antibodies, John Wiley and Sons, New York, 1998).

[0134] Other antibodies that may be useful in treating cancer include those disclosed in Tong et al., Molecules 2021, 26, 5847, https: / / doi.org / 10.3390 / molecules26195847, and Coats et al., Clin Cancer Res 2019;25:5441-8, the entire contents of which are incorporated by reference for purposes of defining antibodies.

[0135] In one embodiment, the antibody is selected from the group consisting of gemtuzumab, brentuximab, ado-trastuzumab, fam-trastuzumab, inotuzumab, polatuzumab, acituzumabab, sacituzumab, belantamab, ioncastuximab, tisotumab, indatuximab, naratuximab, and depatuximab.

[0136] In another embodiment, the antibody is sacituzumab. Alternatively, the antibody is ifinatamab.

[0137] Preferably, the antibody is selected from the group consisting of trastuzumab, rituximab, sacituzumab, and ifinatamab.

[0138] In one embodiment, the antibody is a humanized antibody, a chimeric antibody, a human antibody, or an antibody fragment.

[0139] In one embodiment, the antibody binds to HER2. In one embodiment, the antibody is trastuzumab, pertuzumab, margetuximab, ertumaxomab, MM-111, HER2Bi-aATCs, MCLA-128, ZW25, MDX-210, ado-trastuzumab, and fam-trastuzumab. Preferably, the antibody is trastuzumab. In one embodiment, the antibody has six CDRs of trastuzumab. Trastuzumab comprises a heavy chain of SEQ ID NO: 2 and a light chain of SEQ ID NO: 1.

[0140] In one embodiment, the antibody binds to CD20. Preferably, the antibody is rituximab. In one embodiment, the antibody has six CDRs of rituximab. Rituximab comprises a heavy chain of SEQ ID NO: 4 and a light chain of SEQ ID NO: 3.

[0141] In one embodiment, the antibody binds to an antigen selected from the group consisting of: CLDN18, FOLR1, EGFR, Nectin-4, CD22, c-MET, CD19, CEACAM5, mesothelin, PSMA, ROR1, TF, TNF-BCAM, TROP2, VTCN1, 5T4 carcinoembryonic antigen, AXL, CD276, CD30, CD38, IL3RA, NaPi2b, BRAF, Cadherin-6, CD37, CD70, DLK1, ENPP3, EpCAM, HER3, LRRC15, PD-L1, PTK7, STING, TEM1, TOP1, and VEGF.

[0142] In one embodiment, the antibody binds to a member of claudin 18, specifically claudin 18.2, and for example, an ADC comprising the antibody may be used in the treatment of gastric cancer. In one embodiment, the cancer expresses one or more members of claudin 18, specifically claudin 18.2.

[0143] In one embodiment, the antibody binds to a folate receptor, and for example, an ADC comprising the antibody may be used to treat ovarian cancer and endometrial cancer. In this embodiment, the antibody is preferably mirvetuximab. In one embodiment, the cancer expresses a folate receptor.

[0144] In one embodiment, the antibody binds to an EGF receptor or a member of the EGF receptor family, and for example, an ADC comprising the antibody may be for use in treating multiple types of epithelial cell cancer. In one embodiment, the cancer expresses an EGF receptor.

[0145] In one embodiment, the antibody binds to Nectin 4, and for example, an ADC comprising the antibody may be for use in treating urothelial cancer, bladder cancer, pancreatic cancer, triple-negative breast cancer, non-small cell lung cancer, gastric cancer, esophageal cancer, or ovarian cancer. In this embodiment, the antibody is preferably enfortumab. In one embodiment, the cancer expresses Nectin 4.

[0146] In one embodiment, the antibody binds to CD22, and for example, an ADC comprising the antibody can be for use in treating non-Hodgkin's lymphoma. In one embodiment, the cancer expresses CD22.

[0147] In one embodiment, the antibody binds to c-MET, e.g., an ADC comprising the antibody may be for use in treating non-small cell lung cancer (NSCLC). In one embodiment, the cancer expresses c-MET.

[0148] In one embodiment, the antibody binds to Trop-2, and for example, an ADC comprising the antibody may be used to treat metastatic triple-negative breast cancer and metastatic urothelial carcinoma. In this embodiment, the antibody is preferably sacituzumab. In one embodiment, the cancer expresses Trop-2. Sacituzumab comprises a heavy chain of SEQ ID NO: 8 and a light chain of SEQ ID NO: 7.

[0149] In one embodiment, the antibody binds to tissue factor, and for example, an ADC comprising the antibody may be for use in treating previously treated recurrent or metastatic cervical cancer. In this embodiment, preferably, the antibody is tisotumab. In one embodiment, the cancer expresses tissue factor.

[0150] In one embodiment, the antibody binds to CD276 (B7-H3), and for example, an ADC comprising the antibody may be used in the treatment of prostate cancer. In this embodiment, the antibody is preferably ifinatamab. In one embodiment, the cancer expresses CD276 (B7-H3). Ifinatamab comprises a heavy chain of SEQ ID NO: 6 and a light chain of SEQ ID NO: 5.

[0151] In one embodiment, the antibody binds to receptor tyrosine kinase-like orphan receptor 1 (ROR1), e.g., an ADC comprising the antibody may be for use in treating various hematological tumors and solid malignancies. In one embodiment, the cancer expresses tyrosine kinase-like orphan receptor 1 (ROR1).

[0152] In one embodiment, the antibody binds to BCMA, for example, an ADC comprising the antibody may be for use in treating multiple myeloma. In one embodiment, the cancer expresses BCMA.

[0153] In one embodiment, the antibody binds to PSMA, and for example, an ADC comprising the antibody can be used to treat prostate cancer. In one embodiment, the cancer expresses PSMA.

[0154] In one embodiment, the antibody binds to CEACAM5, and for example, an ADC comprising the antibody can be used to treat advanced non-squamous non-small cell lung cancer (NSCLC). In one embodiment, the cancer expresses CEACAM5.

[0155] In one embodiment, the antibody binds to mesothelin, and for example, an ADC comprising the antibody can be for use in treating mesothelioma, ovarian tumor, pancreatic tumor, gastric tumor, and non-small cell lung tumor. In one embodiment, the cancer expresses mesothelin.

[0156] In one embodiment, the antibody binds to AXL, and for example, an ADC comprising the antibody can be used to treat multiple solid tumors, including non-small cell lung cancer (NSCLC) and triple-negative breast cancer (TNBC). In this embodiment, preferably, the antibody is enapotamab. In one embodiment, the cancer expresses AXL.

[0157] (drug load) Drug loading (referred to as the variable "p") is the average number of NMT inhibitors per antibody. When the compounds of the present invention are conjugated to cysteine ​​residues, drug loading can range from 1 to 10 NMT inhibitors per antibody, i.e., in this case, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 NMT inhibitors are covalently attached to the antibody. Conjugate compositions include a collection of antibodies conjugated with 1 to 10 different NMT inhibitors.

[0158] The average number of NMT inhibitors per antibody in preparations of ADCs from conjugation reactions can be characterized by conventional means, such as UV, reverse-phase HPLC, HIC, mass spectrometry, ELISA assays, and electrophoresis. The quantitative distribution of ADCs with respect to p can also be determined. ELISA can determine the average p value for a particular preparation of ADCs (Hamblett et al. (2004) Clin. Cancer Res. 10:7063-7070; Sanderson et al. (2005) Clin. Cancer Res. 11:843-852). However, due to the detection limits of antibody-antigen binding and ELISA, the distribution of p(NMT inhibitor) values ​​cannot be discerned. Furthermore, ELISA assays for detecting ADCs do not determine where the drug moiety is attached to the antibody, e.g., the heavy or light chain fragment, or the specific amino acid residue. In some cases, separation, purification, and characterization of a homogeneous ADC with a specific p value from ADCs loaded with other NMT inhibitors can be achieved by means such as reverse-phase HPLC or electrophoresis. Such techniques are also applicable to other types of conjugates.

[0159] For some ADCs, p may be limited by the number of binding sites on the antibody. For example, an antibody may have only one or a few cysteine ​​thiol groups, or only one or a few sufficiently reactive thiol groups to which a linker can be attached. Higher drug loading, e.g., p>5, may result in aggregation, insolubility, toxicity, or loss of cell permeability of certain ADCs.

[0160] Typically, less than the theoretical maximum amount of NMT inhibitor is conjugated to an antibody during the conjugation reaction. Antibodies may contain many lysine residues that do not react with, for example, an NMT inhibitor-linker intermediate or linker reagent. Only the most reactive lysine groups can react with an amine-reactive linker reagent. Also, only the most reactive cysteine ​​thiol groups (e.g., sulfhydryls) can react with a thiol-reactive linker reagent. Typically, antibodies do not contain many, if any, free and reactive cysteine ​​thiol groups that can be linked to an NMT inhibitor. Most cysteine ​​thiol residues in a compound's antibody exist as disulfide bridges and must be reduced with a reducing agent such as dithiothreitol (DTT) or TCEP under partial or complete reducing conditions. The loading of the ADC can be controlled in several different ways, including: (i) limiting the molar excess of the antibody-reactive NMT inhibitor-linker intermediate or linker reagent; (ii) limiting the time or temperature of the conjugation reaction; and (iii) partial or limited reduction conditions of the cysteine ​​thiol modification.

[0161] Certain antibodies have reducible interchain disulfides, i.e., cysteine ​​bridges. Antibodies can be made reactive for conjugation with linker reagents by treatment with a reducing agent such as DTT (dithiothreitol). Thus, each cysteine ​​bridge theoretically forms two reactive thiol nucleophiles. Additional nucleophilic groups can be introduced into antibodies by reacting lysines with 2-iminothiolane (Traut's reagent), resulting in the conversion of amines to thiols. Reactive thiol groups can be introduced into antibodies (or fragments thereof) by modifying one, two, three, four, or more cysteine ​​residues (e.g., preparing mutant antibodies containing one or more non-natural cysteine ​​amino acid residues). US 7,521,541 teaches the modification of antibodies by introducing reactive cysteine ​​amino acids. Cysteine ​​amino acids can be engineered into antibodies at reactive sites that do not form intrachain or intermolecular disulfide bridges (Junutula et al., 2008, Nature Biotech., 26(8):925-932; Dornan et al., (2009) Blood 114(13):2721-2729; US 7521541; US ​​7723485; WO2009 / 052249). The engineered cysteine ​​thiol can react with a linker reagent bearing a thiol-reactive electrophilic group, such as a maleimide or an α-haloamide, or a drug-linker reagent of the present invention to form an ADC bearing a cysteine-engineered antibody and an NMT inhibitor moiety. In this way, the location of the NMT inhibitor moiety can be designed, controlled, and identified. The engineered cysteine ​​thiol group typically reacts with thiol-reactive linker reagents or drug-linker reagents in high yield, allowing for controlled NMT inhibitor loading. When an IgG antibody is engineered to incorporate a cysteine ​​amino acid by substitution at a single site on the heavy or light chain, resulting in two new cysteines on a symmetric antibody, an NMT inhibitor loading approaching 2 can be achieved with near-uniformity in the resulting ADC.

[0162] Alternatively, site-specific conjugation can be achieved by modifying an antibody to contain unnatural amino acids in its heavy and / or light chains, as described in Axup et al. ((2012), Proc Natl Acad Sci USA. 109(40):16101-16116). Unnatural amino acids offer the added advantage that orthogonal chemistries can be designed to couple linker reagents and NMT inhibitors.

[0163] When multiple nucleophilic or electrophilic groups on an antibody are reacted with an NMT inhibitor-linker intermediate, or a linker reagent followed by an NMT inhibitor, the resulting product is a mixture of ADC compounds with a distribution of NMT inhibitors attached to the antibody, e.g., 1, 2, 3, etc. Liquid chromatography methods such as polymer reversed-phase (PLRP) and hydrophobic interaction (HIC) can separate compounds in a mixture by NMT inhibitor loading value. Although preparations of ADCs with a single NMT inhibitor loading value (p) can be isolated, these single-loading value ADCs may still be heterogeneous mixtures because the NMT inhibitors may be attached via linkers at different sites on the antibody.

[0164] Thus, the ADCs of the invention include mixtures of ADC compounds in which the antibody bears one or more NMT inhibitor moieties and in which the NMT inhibitor moieties may be attached to the antibody at different amino acid residues.

[0165] In one embodiment, the drug loading (p) of NMT inhibitors on the antibody is 1 to 10 NMT inhibitors per antibody. Preferably, the drug loading (p) is 2 to 6, 4 to 6, 8 to 10, or 6 to 8 NMT inhibitors per antibody.

[0166] In one embodiment, the ADC of the invention has the following formula: [ka] (wherein Ab is an antibody as defined herein, and p is as defined herein). Preferably, the ADC of the invention or a salt thereof is linked to the antibody via a sulfhydryl group in the side chain of a cysteine ​​amino acid on the antibody. Preferably, the antibody is trastuzumab or rituximab, particularly trastuzumab. Alternatively, the antibody is sacituzumab. Alternatively, the antibody is ifinatamab. Preferably, p is 1 to 10, for example, p is 2 to 6, 4 to 6, 8 to 10, or 6 to 8. Most preferably, p is about 5, for example, 5.

[0167] In one embodiment, the ADC of the invention has the following formula: [ka] (wherein Ab is an antibody as defined herein, and p is as defined herein). Preferably, the ADC of the invention or a salt thereof is linked to the antibody via a sulfhydryl group in the side chain of a cysteine ​​amino acid on the antibody. Preferably, the antibody is trastuzumab or rituximab, particularly trastuzumab. Alternatively, the antibody is sacituzumab. Alternatively, the antibody is ifinatamab. Preferably, p is 1 to 10, for example, p is 2 to 6, 4 to 6, 8 to 10, or 6 to 8. Most preferably, p is about 5, for example, 5.

[0168] In one embodiment, the ADC of the invention has the following formula: [ka] (wherein Ab is an antibody as defined herein, and p is as defined herein). Preferably, the ADC of the invention or a salt thereof is linked to the antibody via a sulfhydryl group in the side chain of a cysteine ​​amino acid on the antibody. Preferably, the antibody is trastuzumab or rituximab, particularly trastuzumab. Preferably, p is 1 to 10, for example, p is 2 to 6, 4 to 6, 8 to 10, or 6 to 8. Most preferably, p is about 5, for example, 5.

[0169] In one embodiment, the ADC of the invention has the following formula: [ka] (wherein Ab is an antibody as defined herein, and p is as defined herein). Preferably, the ADC of the invention or a salt thereof is linked to the antibody via a sulfhydryl group in the side chain of a cysteine ​​amino acid on the antibody. Preferably, the antibody is trastuzumab or rituximab, particularly trastuzumab. Alternatively, the antibody is sacituzumab. Alternatively, the antibody is ifinatamab. Preferably, p is 1 to 10, for example, p is 2 to 6, 4 to 6, 8 to 10, or 6 to 8. Most preferably, p is about 5, for example, 5.

[0170] In one embodiment, the ADC of the invention has the following formula: [ka] (wherein Ab is an antibody as defined herein, and p is as defined herein). Preferably, the ADC of the invention or a salt thereof is linked to the antibody via a sulfhydryl group in the side chain of a cysteine ​​amino acid on the antibody. Preferably, the antibody is trastuzumab or rituximab, particularly trastuzumab. Alternatively, the antibody is sacituzumab. Alternatively, the antibody is ifinatamab. Preferably, p is 1 to 10, for example, p is 2 to 6, 4 to 6, 8 to 10, or 6 to 8. Most preferably, p is about 5, for example, 5.

[0171] In one embodiment, the ADC of the invention has the following formula: [ka] (wherein Ab is an antibody as defined herein, and p is as defined herein). NMT is an NMT inhibitor. Preferably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Preferably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof. Preferably, the ADC of the present invention or a salt thereof is linked to the antibody via a sulfhydryl group in the side chain of a cysteine ​​amino acid on the antibody. Preferably, the antibody is trastuzumab or rituximab, particularly trastuzumab. Alternatively, the antibody is sacituzumab. Alternatively, the antibody is ifinatamab. Preferably, p is 1 to 10, for example, p is 2 to 6, 4 to 6, 8 to 10, or 6 to 8. Most preferably, p is about 5, for example, 5.

[0172] (drug conjugates) The ADCs of the invention can be prepared using a drug conjugate, or a salt and / or solvate thereof, which is then covalently bound to an antibody. Thus, in one embodiment, a drug conjugate, or a salt and / or solvate thereof, is provided that includes a group capable of forming a covalent bond with a chain terminus (e.g., N-terminus) or a functional group, e.g., a sulfhydryl group, on an amino acid side chain of an antibody.

[0173] In one embodiment, the drug conjugate has the formula: A a -W w -Y y -NMT(Formula (X)) (In the formula: A, if present, is a First Stretcher unit that contains a group that can form a covalent bond with a chain terminus (e.g., N-terminus) or a functional group, e.g., a sulfhydryl group, on an amino acid side chain of an antibody; NMT is an NMT inhibitor as defined above; and a, W, w, Y, and y are as defined above. or a salt and / or solvate thereof.

[0174] It will be understood that the preferences and embodiments disclosed herein with respect to the ADC of formula (VII) apply equally to the drug conjugate of formula (X).

[0175] The NMT inhibitor forms a covalent bond with the second Stretcher unit (Y), if present, or with the amino acid unit or glucuronide unit (W), if Y is absent. Preferably, the functional group on the NMT inhibitor is amino or alcohol, e.g., amino. When the NMT inhibitor forms a covalent bond via an amino group, the amino group must have an available hydrogen atom to allow reaction with the corresponding functional group (e.g., carbonyl group) in the linker, if present, specifically in the second Stretcher unit (Y), or with the amino acid unit or glucuronide unit (W), if Y is absent; i.e., the amino group cannot be tertiary.

[0176] In one embodiment, the group capable of forming a covalent bond with the chain terminus (eg, N-terminus) or functional group on an amino acid side chain of an antibody is a maleimide moiety.

[0177] In one embodiment, A is [ka] (where, [ka] represents the point of attachment to W) :It is.

[0178] In one embodiment, the drug conjugate is a compound of formula (DC-1): [ka] or a salt and / or solvate thereof, wherein: [ka] is an NMT inhibitor. Preferably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Preferably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof.

[0179] As used herein, " [ka] It will be understood by one of ordinary skill in the art that the phrase "is an NMT inhibitor" refers to the moiety that remains after an NMT inhibitor, e.g., an NMT inhibitor that includes a suitable functional group for attachment to a linker, such as an amino group (which includes a hydrogen atom) or an alcohol (-OH), reacts with a suitable functional group on the linker, e.g., a carbonyl group, thereby forming a linker-NMT inhibitor covalent bond.

[0180] When the NMT inhibitor is a compound of formula (VI), [ka] teeth, [ka] (wherein all variables are as defined above; wherein one group among R5 and R6 is absent so that the N atom can form a covalent bond with the linker. : means

[0181] Similarly, when the NMT inhibitor is a compound of formula (VIA), [ka] teeth, [ka] (wherein all variables are as defined above; wherein one group among R5 and R6 is absent so that the N atom can form a covalent bond with the linker. : means

[0182] In one embodiment, the drug conjugate has the formula (DC-2): [ka] or a salt and / or solvate thereof, wherein: [ka] is an NMT inhibitor. Preferably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Preferably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof.

[0183] In one embodiment, the drug conjugate is a compound of formula (DC-5): [ka] or a salt and / or solvate thereof, wherein: [ka] is an NMT inhibitor. Preferably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Preferably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof.

[0184] In one embodiment, the drug conjugate is a compound of formula (DC-6): [ka] or a salt and / or solvate thereof, wherein: [ka] is an NMT inhibitor. Preferably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Preferably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof.

[0185] In one embodiment, the drug conjugate is (1S,2R,3S,4R,5R)-5-(4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-[3-(3-{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy]ethoxy}propanamido)propanamido]phenoxy)-3,4-dihydroxy-2-methylcyclohexane-1-carboxylic acid: [ka] or a salt and / or solvate thereof.

[0186] In one embodiment, the drug conjugate is {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido]-3-methylbutanamido]pentanamido]phenyl}methyl N-{[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}-N-methylcarbamate: [ka] or a salt and / or solvate thereof.

[0187] In one embodiment, the drug conjugate is (1S,2R,3S,4R,5R)-5-(4-{[({[6-(3,4-difluoro-2-{2-[3-(2-hydroxypropan-2-yl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-[3-(3-{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy]ethoxy}propanamido)propanamido]phenoxy)-3,4-dihydroxy-2-methylcyclohexane-1-carboxylic acid: [ka] or a salt and / or solvate thereof.

[0188] In one embodiment, the drug conjugate is N-[({[(1-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)methyl]carbamoyl}-2-phenylethyl)carbamoyl]methyl}carbamoyl)methyl]-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide: [ka] or a salt and / or solvate thereof.

[0189] The drug conjugate may be provided in the form of a salt. The drug conjugate may be provided in the form of a solvate. The drug conjugate may be provided in the form of a solvate of a salt. Preferably, the drug conjugate is provided.

[0190] The present invention also provides intermediate compounds that are useful in the preparation of drug conjugates.

[0191] In one embodiment, a compound of formula (ADC-I): [ka] (wherein NMT is an NMT inhibitor) or a salt and / or solvate thereof is provided. Preferably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Preferably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof. Preferably, the NMT inhibitor is 1-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}-2,2-dimethylpropan-1-ol or a salt thereof.

[0192] In one embodiment, a compound of Formula (ADC-II): [ka] or a salt and / or solvate thereof, wherein NMT is an NMT inhibitor. Preferably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Preferably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof. Preferably, the NMT inhibitor is 1-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}-2,2-dimethylpropan-1-ol or a salt thereof.

[0193] In one embodiment, the present invention provides {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide]-3- methylbutanamido]pentanamido]phenyl}methyl 4-nitrophenyl carbonate; (1S,2R,3S,4R,5R)-5-[2-(3-{[(9H-fluoren-9-yloxy)carbonyl]amino}propanamido)-4-({[(4-nitrophenoxy)carbonyl]oxy}methyl)phenoxy]-2,3,4-trihydroxycyclohexane-1-carboxylic acid; (1S,2R,3S,4R,5R)-5-(4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-(3-{[(9H-fluoren-9-yloxy)carbonyl]amino}propanamido)phenoxy)-2,3,4-trihydroxycyclohexane-1-carboxylic acid; and (1S,2R,3S,4R,5R)-5-[2-(3-aminopropanamido)-4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}phenoxy]-2,3,4-trihydroxycyclohexane-1-carboxylic acid or a salt and / or solvate of any one thereof.

[0194] It is understood that for use in medicine, salts of the ADCs of the invention should be pharmaceutically acceptable. Non-pharmaceutically acceptable salts of the ADCs of the invention may be useful in other contexts, such as during the preparation of the ADCs of the invention. Suitable pharmaceutically acceptable salts will be apparent to those skilled in the art. Pharmaceutically acceptable salts include those described in Berge et al. (1977). Such pharmaceutically acceptable salts include acid and base addition salts. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, or phosphoric acid, and organic acids, such as succinic acid, maleic acid, acetic acid, fumaric acid, citric acid, tartaric acid, benzoic acid, p-toluenesulfonic acid, methanesulfonic acid, or naphthalenesulfonic acid. Other salts, such as oxalates or formates, may be used, for example, in the isolation of the ADCs of the invention, and are within the scope of the invention.

[0195] Some of the ADCs of the invention may form acid addition salts with one or more equivalents of the acid, and the present invention includes within its scope all possible stoichiometric and non-stoichiometric forms.

[0196] It should be understood that the present invention encompasses all isomers of the ADCs of the invention, including all geometric, tautomeric, and optical forms, and mixtures thereof (e.g., racemic mixtures). If additional chiral centers are present in the ADCs of the invention, the invention includes within its scope all possible diastereoisomers, including mixtures thereof. The different isomeric forms may be separated or resolved one from the other by conventional methods, or any given isomer may be obtained by conventional synthetic methods or by stereospecific or asymmetric syntheses.

[0197] The present disclosure includes all isotopic forms of the ADCs of the invention provided herein, whether (i) in a form in which all atoms of a given atomic number have a mass number (or mixture of mass numbers) that is predominant in nature (referred to herein as a "natural isotopic form"), or (ii) in a form in which one or more atoms have been replaced by an atom having the same atomic number but a mass number that differs from the mass number of the atom that is predominant in nature (referred to herein as a "non-natural variant isotopic form"). It is understood that atoms may naturally exist as a mixture of mass numbers. The term "non-natural variant isotopic form" also includes embodiments in which the proportion of atoms of a given atomic number that have mass numbers that are less commonly found in nature (referred to herein as "uncommon isotopes") is increased relative to the proportion occurring in nature, e.g., to a level of >20%, >50%, >75%, >90%, >95%, or >99% of the number of atoms of that atomic number (the latter embodiments are referred to as "isotopically enriched variant forms"). The term "non-naturally occurring variant isotopic form" also includes embodiments in which the proportion of an uncommon isotope is reduced compared to the proportion occurring in nature. Isotopic forms can include radioactive forms (i.e., those incorporating a radioactive isotope) and non-radioactive forms. Radioactive forms are usually isotopically enriched variant forms.

[0198] Thus, non-naturally occurring variant isotopic forms of the ADCs of the invention may be deuterium ( 2 H or D), carbon-11 ( 11 C), carbon-13( 13 C), carbon-14( 14 C), nitrogen-13( 13 N), nitrogen-15( 15 N), oxygen-15( 15 O), oxygen-17( 17 O), oxygen-18( 18 O), phosphorus-32( 32 P), sulfur-35( 35 S), chlorine-36( 36 Cl), chlorine-37( 37 Cl), fluorine-18( 18 F), iodine-123( 123 I), iodine-125( 125It may contain one or more artificial or uncommon isotopes, such as I), in one or more atoms, or may contain an increased proportion of such isotopes compared to the proportion that predominates in nature.

[0199] Non-natural variant isotopic forms containing radioactive isotopes can be used, for example, for drug and / or substrate tissue distribution studies. The radioactive isotope tritium, i.e. 3 H, and carbon-14, i.e., 14 C are particularly useful for this purpose given their ease of incorporation and ready means of detection. 2 Non-naturally occurring variant isotopic forms incorporating H or D may confer certain therapeutic advantages resulting from greater metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some situations. 11 C. 18 F, 15 O, and 13 Non-natural variant isotopic forms can be prepared incorporating positron emitting isotopes such as N, which are useful in positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0200] In one embodiment, an ADC of the invention or a salt thereof is provided in natural isotopic form.

[0201] In one embodiment, an ADC of the invention or a salt thereof is provided in a non-naturally occurring variant isotopic form. In a specific embodiment, a non-naturally occurring variant isotopic form has a deuterium (i.e., 2 In one embodiment, the atoms of an ADC of the invention or a salt thereof are in an isotopic form that is not radioactive. In one embodiment, one or more atoms of an ADC of the invention or a salt thereof are in an isotopic form that is radioactive. Preferably, the radioisotope is a stable isotope. Preferably, the non-natural variant isotopic form is a pharmaceutically acceptable form.

[0202] In one embodiment, an ADC of the invention or a salt thereof is provided in which a single atom of the compound exists in a non-naturally occurring variant isotopic form. In another embodiment, an ADC of the invention or a salt thereof is provided in which two or more atoms exist in a non-naturally occurring variant isotopic form.

[0203] Non-natural isotopic variant forms can generally be prepared by conventional techniques known to those of skill in the art or by processes similar to those described herein, e.g., those described in the accompanying Examples for preparing natural isotopic forms. Thus, non-natural isotopic variant forms can be prepared by substituting appropriate isotopic variant (or labeling) reagents for the conventional reagents utilized in the Examples. Because the ADCs of the invention are intended for use in pharmaceutical compositions, it will be readily understood that they are each preferably provided in substantially pure form, e.g., at least 60% pure, more suitably at least 75% pure, and preferably at least 85%, particularly at least 98% pure (percentages are on a weight-to-weight basis). Impure preparations of the compounds can be used to prepare purer forms for use in pharmaceutical compositions.

[0204] In general, the ADCs of the present invention can be made according to organic synthesis techniques known to those skilled in the art, as well as by the representative methods shown below, methods in the Examples, and modifications thereof.

[0205] (Uses of the ADC of the present invention) The terms "prophylaxis" or "prevention" are used herein to mean proactive measures and can therefore include preventing symptoms of a disease or disorder in a subject or preventing the recurrence of symptoms of a disease or disorder in an affected subject, and are not limited to complete prevention of affliction.

[0206] The term "treatment" or "treating" as used herein includes controlling, alleviating, reducing or modulating a disease state or its symptoms.

[0207] (Inhibition of human NMT) Inhibition of human NMT has been proposed as a target for treating or preventing various diseases or disorders such as those described above. The present invention proposes an ADC comprising a human NMT inhibitor. As used herein, the term "human NMT inhibitor" is intended to cover any moiety that binds to human NMT. ​​The human NMT is preferably HsNMT1. The inhibitor can act as a competitive inhibitor or a partially competitive inhibitor. The inhibitor can bind to human NMT at the myr-CoA binding pocket or the peptide-binding pocket (or inhibit human NMT via another mechanism). Since the ADC or a pharmaceutically acceptable salt thereof of the present invention comprises an NMT inhibitor that is a human NMT inhibitor, it is believed that after intracellular release of the NMT inhibitor from the ADC of the present invention, the NMT inhibitor will preferably bind to and inhibit human NMT via the peptide-binding pocket.

[0208] Thus, the ADC of the present invention or a pharmaceutically acceptable salt thereof is believed to be useful in the treatment or prevention of a disease or disorder associated with human NMT activity, or in the treatment or prevention of a disease or disorder by targeting human NMT activity, e.g., a hyperproliferative disease such as cancer. Thus, the present invention provides the ADC of the present invention or a pharmaceutically acceptable salt thereof for use as a pharmaceutical.

[0209] Also provided is an ADC of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of a disease or disorder in which inhibition of human NMT provides a therapeutic or preventive effect. In one embodiment, an ADC of the present invention or a pharmaceutically acceptable salt thereof is provided for use in the treatment of a disease or disorder in which inhibition of human NMT provides a therapeutic effect. In one embodiment, an ADC of the present invention or a pharmaceutically acceptable salt thereof is provided for use in the prevention of a disease or disorder in which inhibition of human NMT provides a preventive effect.

[0210] The present invention also provides methods for treating or preventing a disease or disorder in a subject (e.g., a mammal, e.g., a human) in which inhibition of human NMT results in a therapeutic or prophylactic effect in the subject, comprising administering to the subject a therapeutically effective amount of an ADC of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The present invention also provides methods for treating a disease or disorder in a subject (e.g., a mammal, e.g., a human) in which inhibition of human NMT results in a therapeutic effect in the subject, comprising administering to the subject a therapeutically effective amount of an ADC of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The present invention also provides methods for preventing a disease or disorder in a subject (e.g., a mammal, e.g., a human) in which inhibition of human NMT results in a prophylactic effect in the subject, comprising administering to the subject a therapeutically effective amount of an ADC of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0211] The present invention also provides use of an ADC of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment or prevention of a disease or disorder for which inhibition of human NMT provides a therapeutic or preventive effect.The present invention also provides use of an ADC of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a disease or disorder for which inhibition of human NMT provides a therapeutic effect.The present invention also provides use of an ADC of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention of a disease or disorder for which inhibition of human NMT provides a preventive effect.

[0212] (hyperproliferative disorder) Since the ADC of the present invention has cytotoxic activity, the ADC of the present invention or a pharmaceutically acceptable salt thereof is believed to be useful in the treatment or prevention of hyperproliferative disorders.

[0213] Thus, the present invention provides an ADC of the invention or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of a hyperproliferative disorder. In one particularly preferred embodiment, the ADC of the invention or a pharmaceutically acceptable salt thereof is for use in the treatment of a hyperproliferative disorder.

[0214] In one embodiment, the invention provides use of an ADC of the invention, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment or prevention of a hyperproliferative disorder. In one particularly preferred embodiment, the invention provides use of an ADC of the invention, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a hyperproliferative disorder.

[0215] In one embodiment, the invention provides a method of treating or preventing a hyperproliferative disorder in a subject, the method comprising administering a therapeutically effective amount of an ADC of the invention or a pharmaceutically acceptable salt thereof. In one particularly preferred embodiment, the invention provides a method of treating a hyperproliferative disorder in a subject, the method comprising administering a therapeutically effective amount of an ADC of the invention or a pharmaceutically acceptable salt thereof.

[0216] In one embodiment, the hyperproliferative disorder is cancer.

[0217] In one embodiment, the cancer is a hematological malignancy selected from the group consisting of lymphoma (e.g., B-cell lymphoma, particularly a lymphoma selected from the group consisting of high-grade mantle zone lymphoma, follicular lymphoma, plasmablastic lymphoma, diffuse large B-cell lymphoma, and Burkitt's lymphoma), myeloma (e.g., multiple myeloma), leukemia (e.g., a leukemia selected from the group consisting of chronic lymphocytic leukemia, AML, and B-acute lymphocytic leukemia), and melanoma (e.g., a melanoma selected from the group consisting of superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, amelanotic melanoma, and acral lentigo melanoma).

[0218] The cancer may also or alternatively be a solid tumor selected from the group consisting of brain cancer, lung cancer, breast cancer (e.g., triple-negative breast cancer or invasive breast cancer), prostate cancer, ovarian cancer, colorectal (e.g., colon) cancer, gallbladder cancer, kidney cancer, and liver cancer. For example, the cancer may be ovarian serous cystadenocarcinoma, esophageal cancer, lung squamous cell carcinoma, lung adenocarcinoma, bladder urothelial carcinoma, uterine carcinosarcoma, gastric adenocarcinoma, invasive breast cancer, or hepatocellular carcinoma of the liver. In one preferred embodiment, the cancer is breast cancer, for example, triple-negative breast cancer or invasive breast cancer. In one preferred embodiment, the cancer is brain cancer, breast cancer, prostate cancer, colon cancer, gallbladder cancer, or kidney cancer. In some embodiments, the cancer is breast cancer, colon cancer, or gallbladder cancer.

[0219] The cancer may also or alternatively be a blastoma, particularly a neuroblastoma, such as a retinoblastoma, a glioblastoma, a small cell lung cancer, or an astrocytoma.

[0220] In particularly preferred embodiments, the cancer is a hematological malignancy (e.g., lymphoma, particularly B-cell lymphoma (e.g., high-grade mantle zone lymphoma, follicular lymphoma, plasmablastic lymphoma, diffuse large B-cell lymphoma, and Burkitt's lymphoma), myeloma (e.g., multiple myeloma), or leukemia (e.g., chronic lymphocytic leukemia, AML, and B-acute lymphocytic leukemia)), solid tumor (e.g., brain cancer, The cancer may be selected from the group consisting of lung cancer, breast cancer (e.g., triple-negative breast cancer or invasive breast cancer), prostate cancer, ovarian cancer, colorectal (e.g., colon) cancer, gallbladder cancer, kidney cancer, or liver cancer, or neuroblastoma (e.g., retinoblastoma, glioblastoma, small cell lung carcinoma, or astrocytoma)), and melanoma (e.g., superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, amelanotic melanoma, or acral lentigo melanoma).

[0221] In a preferred embodiment, the cancer can be selected from the group consisting of diffuse large B-cell lymphoma, Burkitt's lymphoma, multiple myeloma, neuroblastoma, AML, and B-acute lymphocytic leukemia. In a preferred embodiment, the cancer can be selected from the group consisting of diffuse large B-cell lymphoma, Burkitt's lymphoma, neuroblastoma, AML, B-acute lymphocytic leukemia, and breast cancer. In a preferred embodiment, the cancer can be selected from the group consisting of diffuse large B-cell lymphoma, neuroblastoma, B-acute lymphocytic leukemia, and triple-negative breast cancer. In a preferred embodiment, the cancer can be selected from the group consisting of diffuse large B-cell lymphoma, Burkitt's lymphoma, multiple myeloma, neuroblastoma, AML, B-acute lymphocytic leukemia, and triple-negative breast cancer. In a preferred embodiment, the cancer can be selected from the group consisting of multiple myeloma, neuroblastoma, AML, B-acute lymphocytic leukemia, and triple-negative breast cancer. In a preferred embodiment, the cancer can be selected from the group consisting of multiple myeloma, neuroblastoma, and triple-negative breast cancer.

[0222] In one preferred embodiment, the cancer expresses HER2 protein, i.e., the cancer contains cells with HER2 protein on their cell surface. HER2 protein is overexpressed in various tumors and can be assessed using methods commonly practiced in the art, such as immunohistochemical staining (IHC) to assess HER2 protein overexpression or fluorescence in situ hybridization (FISH) to assess HER2 gene amplification.

[0223] Suitably, the cancer that expresses HER2 protein is selected from the group consisting of lung cancer, urothelial cancer, colorectal cancer, prostate cancer, ovarian cancer, pancreatic cancer, breast cancer, bladder cancer, gastric cancer (e.g., gastric adenocarcinoma), gastrointestinal stromal tumor, cervical cancer, esophageal cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, and penile cancer, e.g., breast cancer.

[0224] In one embodiment, the cancer is HER2-positive breast cancer.

[0225] In one preferred embodiment, the cancer expresses CD20. The expression of CD20 by cancer cells is evaluated using methods commonly practiced in the art, for example, immunohistochemical staining (IHC). Preferably, the cancer that expresses CD20 is lymphoma, for example, high-grade mantle zone lymphoma, follicular lymphoma, plasmablastic lymphoma, diffuse large B-cell lymphoma, or Burkitt's lymphoma), myeloma (for example, multiple myeloma), or leukemia, for example, chronic lymphocytic leukemia, acute myeloid leukemia (AML), or B-acute lymphocytic leukemia.

[0226] In one preferred embodiment, the cancer is a MYC-dependent cancer described in WO2020 / 128475, the entire contents of which are incorporated by reference for purposes of defining MYC-dependent cancers.

[0227] (Combination Therapy) While the ADC of the invention or a pharmaceutically acceptable salt thereof can be used as the sole active ingredient in a pharmaceutical, it is also possible for the ADC of the invention or a pharmaceutically acceptable salt thereof to be used in combination with one or more additional therapeutic agents. Accordingly, the present invention also provides an ADC of the invention or a pharmaceutically acceptable salt thereof together with an additional therapeutic agent. The additional therapeutic components may be for simultaneous, sequential, or separate administration. The present invention also provides a kit of parts comprising: (a) a first pharmaceutical composition comprising an ADC of the invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier; and (b) a second pharmaceutical composition comprising an additional therapeutic agent and a pharmaceutically acceptable carrier. Such additional therapeutic agent may be an ADC of the invention or a pharmaceutically acceptable salt thereof.

[0228] The ADCs of the present invention, or pharmaceutically acceptable salts thereof, can be used in combination with one or more additional therapeutic agents useful for the treatment or prevention of hyperproliferative disorders, such as cancer, or another disease or disorder for which inhibition of human NMT provides a therapeutic or prophylactic effect. The individual components of such combinations can be administered separately at different times during the course of treatment or simultaneously in divided or single combination forms. The present invention should therefore be understood to encompass all such regimes of simultaneous or alternating treatment, and the term "administering" should be interpreted accordingly. It will be understood that the scope of combinations of the ADCs of the present invention, or pharmaceutically acceptable salts thereof, with other therapeutic agents useful for the treatment or prevention of diseases or disorders for which inhibition of human NMT provides a therapeutic or prophylactic effect includes, in principle, any combination with any pharmaceutical composition useful for the treatment or prevention of diseases or disorders for which inhibition of human NMT provides a therapeutic or prophylactic effect.

[0229] When an additional therapeutic agent is utilized in combination with an ADC of the invention or a pharmaceutically acceptable salt thereof, it can be used, for example, in the amount indicated for that agent in the Physicians' Desk Reference (PDR) or as otherwise determined by one of skill in the art. When an ADC of the invention or a pharmaceutically acceptable salt thereof is utilized in combination with one or more additional therapeutic agents, either simultaneously or sequentially, the following combination ratios and dosage ranges are suitable: When combined with an additional therapeutic agent, an ADC of the invention or a pharmaceutically acceptable salt thereof can be utilized, for example, in a weight ratio within the range of about 10:1 to about 1:10 relative to the additional therapeutic agent.

[0230] In one embodiment, when an ADC of the invention or a pharmaceutically acceptable salt thereof is for the treatment or prevention of cancer, the ADC of the invention or a pharmaceutically acceptable salt thereof can be used in combination with one or more additional therapeutic agents, either simultaneously or sequentially, for the treatment or prevention of cancer. More preferably, when an ADC of the invention or a pharmaceutically acceptable salt thereof is for the treatment of cancer, the ADC of the invention or a pharmaceutically acceptable salt thereof can be used in combination with one or more additional therapeutic agents, either simultaneously or sequentially, for the treatment of cancer.

[0231] Suitable, but non-limiting, examples of other therapeutic agents that may be administered in combination with an NMT inhibitor include one or more other chemotherapeutic agents.

[0232] Preferably, the chemotherapeutic agent is fluorouracil (5-FU), pertuzumab, paclitaxel, carboplatin, cisplatin, gemcitabine, capecitabine, irinotecan (CPT-11), paclitaxel, docetaxel, pemetrexed, sorafenib, vinblastine, vinorelbine, everolims, tanespimycin, bevacizumab, oxaliplatin, lapatinib, ado-trastuzumab emtansine (TDM1), or any of the chemotherapeutic agents described in WO 2004 / 022106. The drug is selected from the group consisting of drugs described in US Pat. No. 2003 / 038043, LH-RH analogues (e.g., leuprorelin or goserelin), estramustine phosphate, estrogen antagonists (e.g., tamoxifen or raloxifene), and aromatase inhibitors (e.g., anastrozole, letrozole, or exemestane).

[0233] Such conjoint treatment can be achieved by simultaneous, sequential, or separate administration of an ADC of the invention, or a pharmaceutically acceptable salt thereof, and one or more other therapeutic agents of the treatment. Such combination products can utilize the ADC of the invention, or a pharmaceutically acceptable salt thereof, within any suitable dosage range, e.g., the dosage ranges described herein, and the other pharmaceutically active agent can be within its approved dosage range.

[0234] (Dosage and Formulation) Those skilled in the art will understand that appropriate dosages of the ADCs of the present invention or pharmaceutically acceptable salts thereof, and compositions comprising the ADCs of the present invention or pharmaceutically acceptable salts thereof, may vary from patient to patient. Determining the optimal dosage typically involves balancing the level of therapeutic benefit with any risk or deleterious side effects. The selected dosage level will depend on a variety of factors, including, but not limited to, the activity of the particular ADC, the route of administration, the time of administration, the rate of excretion of the ADC, the duration of treatment, other drugs, ADCs, and / or materials used in combination, the severity of the condition, and the patient's species, sex, age, weight, condition, general health, and medical history. The amount and route of administration of the ADC are ultimately at the discretion of the physician, veterinarian, or clinician, but typically, the dosage will be selected to achieve a local concentration at the site of action that achieves the desired effect without causing significant, harmful, or deleterious side effects.

[0235] Administration can be achieved in one dose, continuously, or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Methods of determining the most effective means and dosage of administration are well known to those skilled in the art and will vary with the formulation used for treatment, the purpose of the treatment, the target cell being treated, and the subject being treated. Single or multiple administrations can be carried out, with the dose level and pattern being selected by the treating physician, veterinarian, or clinician.

[0236] In general, a suitable dose of an ADC ranges from about 100 ng to about 25 mg (more typically, from about 1 μg to about 10 mg) per kilogram of subject body weight per day. When the ADC is a salt, ester, amide, prodrug, or the like, the amount administered is calculated based on the original ADC, and therefore the actual weight used will be increased proportionately.

[0237] In one embodiment, an ADC of the invention or a pharmaceutically acceptable salt thereof is administered to a human patient according to the following dosing regimen: about 100 mg, three times daily. In one embodiment, an ADC of the invention or a pharmaceutically acceptable salt thereof is administered to a human patient according to the following dosing regimen: about 150 mg, twice daily. In one embodiment, an ADC of the invention or a pharmaceutically acceptable salt thereof is administered to a human patient according to the following dosing regimen: about 200 mg, twice daily. In one embodiment, an ADC of the invention or a pharmaceutically acceptable salt thereof is administered to a human patient according to the following dosing regimen: about 50 or about 75 mg, three or four times daily. In one embodiment, an ADC of the invention or a pharmaceutically acceptable salt thereof is administered to a human patient according to the following dosing regimen: about 100 or about 125 mg, twice daily.

[0238] The above dosage amounts can be applied to an effective amount of the ADC of the present invention or a pharmaceutically acceptable salt thereof (comprising an NMT inhibitor and a linker to the antibody) or the NMT inhibitor provided, for example, the amount of the NMT inhibitor that can be released after cleavage of the linker.

[0239] For the prevention or treatment of disease, the appropriate dosage of the ADC of the invention depends on the type of disease being treated, the severity and course of the disease, whether the molecule is administered for prophylactic or therapeutic purposes, as defined above, previous treatments, the patient's clinical history and response to the antibody, and the discretion of the attending physician. The ADC of the invention or a pharmaceutically acceptable salt thereof is preferably administered to a patient at one time or over a series of treatments. For example, an initial candidate dosage for administration to a patient, whether by one or more individual administrations or by continuous infusion, ranges from about 1 μg / kg to 15 mg / kg (e.g., 0.1 to 20 mg / kg) of the ADC of the invention or a pharmaceutically acceptable salt thereof, depending on the type and severity of the disease. Typical daily dosages can range from about 1 μg / kg to 100 mg / kg or more, depending on the factors described above. Exemplary dosages of the ADC of the invention or a pharmaceutically acceptable salt thereof administered to a patient range from about 0.1 mg / kg to about 10 mg / kg of the patient's body weight. For repeated administrations over several days or longer, depending on the condition, treatment is sustained until a desired suppression of disease symptoms occurs. An exemplary dosing regimen involves administering an initial loading dose of about 4 mg / kg, followed by administration of additional doses of the ADC of the invention or a pharmaceutically acceptable salt thereof every one, two, or three weeks. Other dosage regimens may also be useful. The progress of this therapy is easily monitored by conventional techniques and assays.

[0240] While it is possible for the active ingredient to be administered alone, it is preferable for it to be present in a pharmaceutical formulation or composition. Accordingly, the invention provides pharmaceutical formulations or compositions comprising an ADC of the invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent, excipient, or carrier (collectively referred to herein as "carrier" materials). Pharmaceutical compositions of the invention can be in the form of a pharmaceutical formulation.

[0241] Thus, in one embodiment, the invention provides a pharmaceutical composition comprising an ADC of the invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0242] In one embodiment, a pharmaceutical composition is provided comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) for use in the treatment or prevention of a disease or disorder described herein. In one embodiment, a pharmaceutical composition is provided comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) for use in the treatment of a disease or disorder described herein. In one embodiment, a pharmaceutical composition is provided comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) for use in the prevention of a disease or disorder described herein.

[0243] In a further embodiment, methods are provided for the treatment or prevention of a disease or disorder described herein, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt). In a further embodiment, methods are provided for the treatment of a disease or disorder described herein, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt). In a further embodiment, methods are provided for the prevention of a disease or disorder described herein, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt). The pharmaceutical compositions of the invention can take the form of pharmaceutical formulations as described below.

[0244] The invention also provides the use of a pharmaceutical composition comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) in the manufacture of a medicament for the treatment or prevention of a disease or disorder described herein.The invention also provides the use of a pharmaceutical composition comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) in the manufacture of a medicament for the treatment of a disease or disorder described herein.The invention also provides the use of a pharmaceutical composition comprising an ADC of the invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) in the manufacture of a medicament for the prevention of a disease or disorder described herein.

[0245] Preferably, the disease or disorder is a hyperproliferative disorder, such as cancer. In one preferred embodiment, the cancer is a cancer that expresses the HER2 protein.

[0246] The ADCs of the present invention or pharmaceutically acceptable salts thereof can be administered by intravenous, cutaneous, intramuscular, or subcutaneous injection or infusion. The active ingredient is in the form of a parenterally acceptable aqueous solution that is pyrogen-free and has suitable pH, isotonicity, and stability. Those skilled in the art will be able to prepare suitable solutions using, for example, isotonic vehicles such as sodium chloride injection, Ringer's injection, lactated Ringer's injection, preservatives, stabilizers, buffers, antioxidants, and / or other additives, which may be included as needed.

[0247] In one embodiment, the pharmaceutical composition further comprises another active agent, eg, another therapeutic or prophylactic agent.

[0248] Suitable carriers, diluents, excipients, etc. can be found in standard pharmaceutical textbooks. See, for example, Handbook of Pharmaceutical Additives, 2nd Edition (M. Ash and I. Ash, eds.), 2001 (Synapse Information Resources, Endicott, New York, USA), Remington's Pharmaceutical Sciences, 20th Edition, Lippincott, Williams & Wilkins Publishing, 2000; and Handbook of Pharmaceutical Excipients, 2nd Edition, 1994.

[0249] Another aspect of the invention relates to a method of making a pharmaceutical composition, comprising mixing at least one [C]-radiolabelled ADC of the invention, or a pharmaceutically acceptable salt thereof, together with one or more other pharmaceutically acceptable ingredients known to those of skill in the art, such as carriers, diluents, excipients, etc. When formulated as discrete units (e.g., tablets, etc.), each unit contains a predetermined amount (dosage) of active compound.

[0250] As used herein, the term "pharmaceutically acceptable" refers to compounds, ingredients, materials, compositions, dosage forms, etc., that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of the subject without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Each carrier, diluent, excipient, etc. must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

[0251] The formulations may be prepared by any method well known in the art of pharmacy. Such methods include the step of bringing into association an ADC of the invention, or a pharmaceutically acceptable salt thereof, with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association an ADC of the invention, or a pharmaceutically acceptable salt thereof, with the carrier (e.g., liquid carrier, finely divided solid carrier, etc.), and then, if necessary, shaping the product.

[0252] Formulations suitable for parenteral administration (e.g., by injection or infusion) include aqueous or non-aqueous, isotonic, pyrogen-free, sterile liquids (e.g., solutions, suspensions) in which the active ingredient is dissolved, suspended, or otherwise provided (e.g., in liposomes or other microparticles). Such liquids may further contain other pharmaceutically acceptable ingredients, such as antioxidants, buffers, preservatives, stabilizers, bacteriostats, suspending agents, thickeners, and solutes that render the formulation isotonic with the blood (or other relevant body fluids) of the intended recipient. Examples of excipients include, for example, water, alcohols, polyols, glycerol, vegetable oils, and the like. Examples of suitable isotonic carriers for use in such formulations include sodium chloride injection, Ringer's solution, or lactated Ringer's injection. Typically, the concentration of the ADC of the invention or a pharmaceutically acceptable salt thereof in the liquid is from about 1 ng / ml to about 10 μg / ml, e.g., from about 10 ng / ml to about 1 μg / ml. The formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.

[0253] The formulations can be prepared to provide fast or slow release; immediate, delayed, timed, or sustained release; or a combination thereof.

[0254] In some embodiments, the ADC of the present invention or a pharmaceutically acceptable salt thereof can be administered orally. Pharmaceutical compositions for oral administration may be in tablet, capsule, powder, or liquid form. Tablets may contain a solid carrier or adjuvant. Liquid pharmaceutical compositions typically contain a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil, or synthetic oil. Physiological saline solution, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol, or polyethylene glycol may also be included. Capsules may contain a solid carrier such as gelatin.

[0255] In one preferred embodiment, administration is by infusion or injection, particularly by bolus injection.

[0256] The ADCs of the invention, or pharmaceutically acceptable salts thereof, are believed to exhibit one or more of the following advantageous properties: in vivo cytotoxic activity, as demonstrated, for example, in the mouse xenograft model of Biological Example 2; and - in vivo tolerability, as shown, for example, in the mouse xenograft models of Biological Examples 2, 3, 10, 11, 12, and 13 and the monkey model of Biological Example 9; - in vitro cytotoxicity against various cancer cell lines, as shown, for example, in Biological Examples 4, 6, 7, and 14; - Improved efficacy against gastric cancer organoids compared to known controls, for example, as shown in Biological Example 8; and - Improved bystander effect, as shown in Biological Example 6.

[0257] Such properties are believed to make the ADCs of the invention, or pharmaceutically acceptable salts thereof, suitable for use in the treatment or prevention (e.g., treatment) of hyperproliferative disorders such as cancer or other diseases or disorders in which inhibition of human NMTs provides a therapeutic or preventative effect. [Example]

[0258] (Example) (abbreviation) [Table 1] TIFF2025530213000075.tif177170

[0259] (Synthesis of Example Compounds) (General experimental details) (LC-MS) Compounds requiring purification under basic conditions were typically purified on an LC-MS system equipped with a YMC Actus Triart C18 5 μm (20 × 250 mm) column or a Gemini NX 5 μm C18 (100 × 30 mm) column using a gradient elution of acetonitrile in water containing 20 mM ammonium bicarbonate (10–45% over 30 min, followed by 95% acetonitrile for 2 min).

[0260] (HPLC) The purity of a compound was determined on a TyeEclipse Extend or XDB 5 μm C18 (150 × 4.6 mm), Xbridge 5 μm C18 (100 × 4.6 mm), Zorbax Extend 5 μm C18 (150 × 4.6 mm), or Shimadzu L Column 2 ODS 5 μm C18 (150 × 4.6 mm) column using a gradient elution of acetonitrile in water containing 10 mM ammonium acetate over 15 min (HPLC B), 17 min (B1), and 18 min (B3).

[0261] The purity of certain compounds was determined by analytical HPLC using a Poroshell 120 2.7 μm EC18 (100 × 4.6 mm), a Luna Omega Polar 3 μm C18 (100 × 4.6 mm), an Xbridge 5 μm C18 (150 × 4.6 mm), or a Sunfire 5 μm C18 (100 × 4.6 mm) column, using a gradient elution of acetonitrile in water containing 0.05% trifluoroacetic acid over 12 min (HPLC A), 14 min (A1), or 17 min (A2), and 16 min (A4).

[0262] The purity of certain compounds was determined by analytical HPLC using a Gemini NX 3 μm C18 (100×4.6 mm) column with gradient elution of acetonitrile in water containing 0.05% formic acid over 16 min (A6).

[0263] (NMR) 1 H NMR and 13 C spectra were recorded at room temperature on 400 MHz and 101 MHz instruments, respectively, and referenced to the residual solvent signal unless otherwise specified. Data are presented as follows: chemical shift in ppm, integral, multiplicity (br = broad, s = singlet, d = doublet, t = triplet, q = quartet, p = quintet, m = multiplet), and coupling constants (in Hz).

[0264] (ADC test method) (SEC-HPLC) Column: TOSOH TSKgel G3000SWXL 7.8 mm x 30 cm 5 μm particles (MERCK 808541) coupled to a security guard column (MERCK 822858) with a GFC3000 4 x 3 mm cartridge (Phenomenex); buffer: 0.2 M phosphate, 0.25 M KCl, 10% IPA; gradient: isocratic 0.5 ml / min at 25 °C. Sample load was approximately 10 μg, and monomer and concentration were determined from the 214 nm signal. Monomer was reported based on peak integration, and [ADC] mg / mL was based on the antibody calibration curve.

[0265] (RP-HPLC for residual NMT inhibitors) Column: Kinetex® 2.6 μm C8 100 Å, LC column 50×4.6 mm (Phenomex 00B-4497-E0); Mobile phase A 0.05% TFA in water; Mobile phase B 0.05% TFA in CAN; Gradient, 2 ml / min at 60° C.: [Table 2]

[0266] 50 μl sample (ADC or PBS / PS20 matrix) + 2 μl 5 M NaCl + 150 μl cold MeOH (from -20°C freezer). Incubated at -20°C for 30 minutes. Centrifuged at 21,000 g for 30 minutes at 4°C. 125 μl of supernatant was extracted and mixed with 125 μl of WFI. 100 μl of this was injected onto the Kinetex column. Data were analyzed at 214 nm, and residual NMT inhibitor in the sample was estimated from an external calibration curve of the relevant NMT inhibitor-linker. Results are expressed as a percentage of free versus bound, using the ADC concentration and calculated DAR to determine the amount of bound NMT inhibitor.

[0267] (HIC-HPLC for calculating the average DAR (drug-antibody ratio)) This method can be used as an alternative to the PLRP-HPLC method for determining the mean DAR.

[0268] Column: TOSOH Butyl-NPR 4.6 mm x 3.5 cm, 2.5 μm particle size (Merck 822855); Mobile phase A: 1.5 M (NH4)2SO4, 25 mM NaPi, pH 6.95 ± 0.05; Mobile phase B: 25 mM NaH2PO4 pH 6.95 ± 0.05 + 25% IPA; Gradient, 0.8 ml / min at 25 °C: [Table 3] Cartridge load: 10 μg and reported results analyzed at 214 nm.

[0269] (RP-HPLC for average DAR calculation) Column—PLRP-S 2.1 mm × 5 cm, 5 μm (Agilent PL1912-1502); Mobile phase A: 0.1% TFA in water; Mobile phase B: 0.1% TFA in acetonitrile; Gradient, 1 mL / min at 80 °C: [Table 4]

[0270] ~10 μg of sample (ADC) + 5 μl of 0.1 M DTT was adjusted to 50 μL with 0.5 M Tris, pH 8.0, and incubated at 37°C for 15 minutes. The sample was then diluted 1:1 with 49% water, 49% acetonitrile, 2% formic acid (+ 50 μL). 20 μL of this solution was then injected onto the RP-HPLC column. Data was analyzed at 214 nm, and the average DAR was calculated.

[0271] (Endotoxin dynamic chromogenic assay) Endotoxin was determined by a kinetic chromogenic LAL assay using the Endosafe PTS endotoxin system. ADC was diluted 10-fold in LAL reagent water. All samples were analyzed on 0.01-1 EU / mL cartridges. EU / mL values ​​were converted to EU / mg by dividing by ADC[P]mg / mL.

[0272] (Preparation of NMT inhibitor 1: 1-(4-(2-(2,3-difluoro-6-(3-((methylamino)methyl)imidazo[1,2-a]pyridin-6-yl)phenoxy)ethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-ol) [ka] (Step 1 - Intermediate (2): 1,5-dimethyl-1H-pyrazole-3-carboxylic acid) Procedure: To a solution of ethyl 1,5-dimethyl-1H-pyrazole-3-carboxylate (intermediate (1)) (20.0 g, 118.984 mmol) in THF:water (4:1) (280 ml, 70 ml) was added ethanol (0.4 ml) and LiOH.HO (9.985 g, 237.968 mmol) at room temperature. The resulting mixture was stirred at room temperature for 16 hours. TLC / LCMS showed complete consumption of SM. The reaction mixture was acidified with 3N HCl solution (pH approx. 2) at 0°C and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated to give 1,5-dimethyl-1H-pyrazole-3-carboxylic acid (2) as a pale yellow solid (16 g, 99%). [ka]

[0273] (Step 2 - Intermediate (3): N-Methoxy-N,1,5-trimethyl-1H-pyrazole-3-carboxamide) Procedure: To a stirred solution of 1,5-dimethyl-1H-pyrazole-3-carboxylic acid (intermediate (2)) (16.6 g, 118.571 mmol) in tetrahydrofuran (350.0 ml) was added N,O-dimethylhydroxylamine hydrochloride (17.34 g, 177.857 mmol). Triethylamine (82.633 ml, 592.857 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (34.095 g, 177.857 mmol), and 1-hydroxybenzotriazole (24.032 g, 177.857 mmol) were added and the reaction mixture was stirred at RT for 16 hours. TLC showed the formation of the product. The reaction was washed with sodium bicarbonate solution and extracted with ethyl acetate. The organic layer was washed with water, brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude was purified by combiflash with 5% MeOH in DCM to give N-methoxy-N,1,5-trimethyl-1H-pyrazole-3-carboxamide (3) as a pale yellow solid (15.0 g, 69.05%). [ka] LCMS (NH4Oac:CAN): M+H = 184, Rt = 2.17 min, 5 min run.

[0274] (Step 3 - Intermediate (4): 1-(1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one) Procedure: A stirred solution of N-methoxy-N,1,5-trimethyl-1H-pyrazole-3-carboxamide (intermediate (3)) (15.0 g, 81.922 mmol) in tetrahydrofuran (150.0 ml) was cooled to −50° C., and t-butyllithium (1.7 M in pentane) (96.379 ml, 163.844 mmol) was added at −50° C. The reaction mixture was then stirred at −50° C. for 2 hours. TLC showed the formation of the product, and the reaction mixture was quenched with saturated NH4Cl solution. The reaction mixture was diluted with ethyl acetate and washed with water and brine solution. The organic layer was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude was purified by combiflash chromatography using 5% MeOH in DCM to give 1-(1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (4) as a pale yellow solid (6.0 g, 40.63%). [ka] LCMS(HCOOH:CAN): M+H=181, Rt=1.86 minutes, run for 3 minutes.

[0275] (Step 4-Intermediate (5): 1-(4-bromo-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one) Procedure: To a solution of 1-(1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (intermediate (4)) (6.0 g, 33.309 mmol) in acetonitrile (100.0 ml) was added N-bromosuccinimide (6.191 g, 34.975 mmol) portionwise under ice-cooled conditions. The resulting reaction mixture was stirred at RT for 16 hours. TLC and LCMS showed the formation of the product. The reaction mixture was then diluted with ethyl acetate and washed with saturated NaHCO3 solution, water, and brine solution. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to give 1-(4-bromo-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (5) as a yellow solid compound (8.0 g, 92.68%), which was used in the next step without purification. [ka] LCMS (NH4Oac:CAN): M+H = 259, Rt = 3.59 min, 5 min run.

[0276] (Step 5-Intermediate (6): 1-(1,5-dimethyl-4-vinyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one) Procedure: To a solution of 1-(1,5-dimethyl-4-vinyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (intermediate (5)) (7.0 g, 27.129 mmol) in anhydrous N,N-dimethylformamide (100.0 ml) was added tributylvinyltin (17.2 ml, 54.257 mmol) at room temperature. Argon was then purged through the reaction mixture for 15 minutes, and Pd(PPh3)4 (3.133 g, 2.713 mmol) was added. The reaction mixture was stirred at 110°C for 16 hours. TLC showed the consumption of the starting material and the formation of the desired product. The reaction mixture was then diluted with ethyl acetate, washed with potassium fluoride solution, and the precipitate was filtered through a cinnamon, washed with water and brine, dried over sodium sulfate, and concentrated. The crude material was purified by column chromatography (100-200) in 10% ethyl acetate-hexane to give 1-(1,5-dimethyl-4-vinyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (6) (5.0 g, 89.35%). [ka] LCMS(HCOOH:CAN): M+H=207, Rt=2.19 minutes, run for 3 minutes.

[0277] (Step 6 - Intermediate (7): 2-(1,5-dimethyl-3-pivaloyl-1H-pyrazol-4-yl)acetaldehyde) Procedure: To a solution of 1-(1,5-dimethyl-4-vinyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (intermediate (6)) (4.1 g, 19.903 mmol) and (diacetoxyiodo)benzene (6.729 g, 20.898 mmol) in acetonitrile (60.0 ml) was added 5% sulfuric acid (3.525 ml) dropwise at −30° C. The mixture was stirred at −30° C. for 1 hour. After completion of the reaction, the residue was treated with ethyl acetate and washed with saturated sodium bicarbonate solution, water, and brine solution. The aqueous layer was back-extracted with ethyl acetate, and the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give 2-(1,5-dimethyl-3-pivaloyl-1H-pyrazol-4-yl)acetaldehyde (7) (2.7 g, 61.03%). This fraction was then used in the next step without purification. [ka] LCMS (NH4Oac:CAN): M+H = 223, Rt = 1.86 min, 3 min run.

[0278] (Step 7-Intermediate (8): 1-(4-(2-hydroxyethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one) Procedure: To a solution of 2-(1,5-dimethyl-3-pivaloyl-1H-pyrazol-4-yl)acetaldehyde (intermediate (7)) (2.7 g, 12.162 mmol) in ethanol (60.0 ml) was added sodium borohydride (0.460 g, 12.162 mmol) portionwise under ice-cooled conditions. The reaction mixture was stirred at 0° C. for 30 minutes. After completion of the reaction, the mixture was quenched with sodium bicarbonate solution, diluted with ethyl acetate, washed with water, brine, and concentrated in vacuo to give the crude product. The batch was purified by combiflash using 2% MeOH in DCM to give 1-(4-(2-hydroxyethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (8) (2.1 g, 76.98%) as a colorless oil. [ka] LCMS(HCOOH:CAN): M+H=225, Rt=1.81 minutes, run for 3 minutes.

[0279] (Step 8 - Intermediate (9): tert-butyl ((6-(2-(2-(1,5-dimethyl-3-pivaloyl-1H-pyrazol-4-yl)ethoxy)-3,4-difluorophenyl)imidazo[1,2-a]pyridin-3-yl)methyl)(methyl)carbamate) Procedure: To a stirred solution of 1-(4-(2-hydroxyethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (intermediate (8)) (2.3 g, 10.268 mmol) and tert-butyl ((6-(3,4-difluoro-2-hydroxyphenyl)imidazo[1,2-a]pyridyl-3-yl)methyl)(methyl)carbamate (3.994 g, 10.268 mmol) in toluene (40.0 ml) was added CMBP (5.382 ml, 20.536 mmol) at room temperature, and the reaction mixture was stirred at 110° C. for 16 hours. TLC and LCMS indicated the formation of the product, and the reaction mixture was diluted with ethyl acetate, washed with water, brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude material was purified by combiflash with 5% MeOH-DCM to give tert-butyl ((6-(2-(2-(1,5-dimethyl-3-pivaloyl-1H-pyrazol-4-yl)ethoxy)-3,4-difluorophenyl)imidazo[1,2-a]pyridin-3-yl)methyl)(methyl)carbamate (9) as a brown sticky gum (3.0 g, 49.05%). [ka] LCMS(HCOOH:CAN): M+H=596, Rt=1.75 minutes, run for 5 minutes.

[0280] (Step 9 - Intermediate (10): tert-butyl((6-(3,4-difluoro-2-(2-(3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl)ethoxy)phenyl)imidazo[1,2-a]pyridin-3-yl)methyl)(methyl)carbamate) Procedure: To a solution of tert-butyl ((6-(2-(2-(1,5-dimethyl-3-pivaloyl-1H-pyrazol-4-yl)ethoxy)-3,4-difluorophenyl)imidazo[1,2-a]pyridyl-3-yl)methyl)(methyl)carbamate (intermediate (9)) (2.5 g, 4.202 mmol) in methanol (25.0 ml) was added lithium borohydride (0.458 g, 21.008 mmol). The mixture was stirred at ambient temperature for 5 hours. After completion of the reaction, the solvent was evaporated, diluted with DCM, and washed with sodium bicarbonate solution, water, and brine. The organic layer was dried over sodium sulfate and concentrated to give the crude product. The crude product was purified by preparative TLC using 5% MeOH in DCM to give tert-butyl ((6-(3,4-difluoro-2-(2-(3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl)ethoxy)phenyl)imidazo[1,2-a]pyridin-3-yl)methyl)(methyl)carbamate (10) (1.9 g, 75.66%). [ka] LCMS (NH4Oac:CAN): M+H = 598, Rt = 3.75 min, run for 5 min.

[0281] (Step 10—NMT Inhibitor 1: 1-(4-(2-(2,3-difluoro-6-(3-((methylamino)methyl)imidazo[1,2-a]pyridin-6-yl)phenoxy)ethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-ol) Procedure: To a solution of tert-butyl ((6-(3,4-difluoro-2-(2-(3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl)ethoxy)phenyl)imidazo[1,2-a]pyridin-3-yl)methyl)(methyl)carbamate (10) (1.2 g, 2.009 mmol) in diethyl ether (10.0 ml) was added 2 M HCl in diethyl ether (40.0 ml) at 0° C. The reaction mixture was stirred at rt for 3 h. TLC and LCMS showed that the starting material had been consumed. The reaction mixture was evaporated under reduced pressure to give the crude product. The crude material was triturated with diethyl ether and lyophilized to give 1-(4-(2-(2,3-difluoro-6-(3-((methylamino)methyl)imidazo[1,2-a]pyridin-6-yl)phenoxy)ethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-ol as a pale yellow solid (HCl salt) (1.04 g, 96.93 mmol, 49%). [ka] LCMS(HCOOH:CAN): M+H=498, Rt=2.54 min, 5 min run; HPLC RT(B3) 8.739 min.

[0282] Using methods similar to those described for NMT inhibitor 1, the following compounds were made: Table 1: NMT inhibitors [Table 5] TIFF2025530213000091.tif249170TIFF2025530213000092.tif218170TIFF2025530213000093.tif234170TIFF20255302130 00094.tif248170TIFF2025530213000095.tif249170TIFF2025530213000096.tif249170TIFF2025530213000097.tif114170

[0283] NMT inhibitors 1-25 were prepared according to the methods disclosed in EP Patent Application No. 22194959.7, the entire contents of which are incorporated by reference for purposes of describing the synthesis and activity of NMT inhibitors.

[0284] (Synthesis of Drug Conjugate 1: {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido]-3-methylbutanamido]pentanamido]phenyl}methyl N-{[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}-N-methylcarbamate:) [ka] {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido]-3-methylbutanamido]pentanamido]phenyl}methyl 4-nitrophenyl carbonate hydrate (Synlett, 2009, No. 18, pp A solution of 12 mg of ADC intermediate 1 (prepared from commercially available N-[(1S)-1-{[(1S)-4-(carbamoylamino)-1-{[4-(hydroxymethyl)phenyl]carbamoyl}butyl]carbamoyl}-2-methylpropyl]-6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamide) in 2 mL of anhydrous DMF was prepared using standard carbonate formation conditions described in 3050-3051. NMT inhibitor 1 (8 mg, 16 mmol) was added, followed by DIEA (8.4 mL) and HOAt (1 mg), and the reaction was stirred at room temperature (22 °C). After 16 h, the mixture was directly purified by RP-HPLC to give {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido]-3-methylbutanamido]pentanamido]phenyl}methyl N-{[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}-N-methylcarbamate (Drug Conjugate 1) was obtained as a white solid (10.5 mg) after lyophilization. Drug Conjugate 1 can be used to prepare an ADC by reaction with an antibody using methods known to those of skill in the art and described herein, for example, the methods described for ADC Example 1 or ADC Example 2.

[0285] Synthesis of Drug Conjugate 2: (1S,2R,3S,4R,5R)-5-(4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-[3-(3-{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy]ethoxy}propanamido)propanamido]phenoxy)-3,4-dihydroxy-2-methylcyclohexane-1-carboxylic acid (as the hydrate) [ka] Step 1: To a solution of (1S,2R,3S,4R,5R)-5-[2-(3-{[(9H-fluoren-9-yloxy)carbonyl]amino}propanamide)-4-({[(4-nitrophenoxy)carbonyl]oxy}methyl)phenoxy]-2,3,4-trihydroxycyclohexane-1-carboxylic acid (ADC intermediate 2, 100 mg, 0.11 mmol) (Bioconjugate Chem., 2006, 17, 831-840) in anhydrous DMF (2 mL) was added NMT inhibitor 1 (50 mg), followed by DIEA (40 mL) and HOAt (3 mg), and the reaction was stirred at room temperature (22 °C). After 16 h, the mixture was directly purified by RP-HPLC to give (1S,2R,3S,4R,5R)-5-(4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-(3-{[(9H-fluoren-9-yloxy)carbonyl]amino}propanamido)phenoxy)-2,3,4-trihydroxycyclohexane-1-carboxylic acid (ADC intermediate 3) as a white solid (107 mg) after lyophilization.

[0286] Step 2: (1S,2R,3S,4R,5R)-5-(4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-(3-{[(9H-fluoren-9-yloxy)carbonyl]amino}propanamido)phenoxy)-2,3,4-trihydroxycyclohexane-1-carboxylic acid (ADC intermediate 3, 105 mg) was dissolved in acetonitrile / water (6 / 4, v / v, 4 mL), and NaOH (1N, aqueous, 0.5 mL) was added dropwise at room temperature. The mixture was stirred at room temperature for 8 hours. HCl (4N in dioxane, 0.1 mL) was added and the mixture was purified by RP-HPLC to give (1S,2R,3S,4R,5R)-5-[2-(3-aminopropanamido)-4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}phenoxy]-2,3,4-trihydroxycyclohexane-1-carboxylic acid (ADC intermediate 4) as a white solid (TFA salt, 42 mg) after lyophilization.

[0287] Step 3: To a solution of (1S,2R,3S,4R,5R)-5-[2-(3-aminopropanamido)-4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}phenoxy]-2,3,4-trihydroxycyclohexane-1-carboxylic acid (ADC intermediate 4, 40 mg) in acetonitrile / water (6 / 4, v / v, 2 mL) was added Mal-PEG2-OSu (15 mg) followed by DIEA (14 mL). The reaction mixture was stirred at room temperature for 1 h and directly purified by RP-HPLC to give (1S,2R,3S,4R,5R)-5-(4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-(3-{[(9H-fluoren-9-yloxy)carbonyl]amino}propanamido)phenoxy)-2,3,4-trihydroxycyclohexane-1-carboxylic acid hydrate (drug conjugate 2) as a white solid (36 mg) after lyophilization.

[0288] (Synthesis of Drug Conjugate 3) [ka] Step 1: To a solution of ADC intermediate 2 (Bioconjugate Chem., 2006, 17, 831-840) in anhydrous DMF, NMT inhibitor 26 was added, followed by DIEA and HOAt, and the reaction was stirred at room temperature (22 °C). After 16 h, the mixture was directly purified by RP-HPLC to give ADC intermediate 5 as a white solid after lyophilization.

[0289] Step 2: ADC intermediate 5 was dissolved in acetonitrile / water (6 / 4, v / v), and NaOH (1N, aqueous) was added dropwise at room temperature. The mixture was stirred at room temperature for 8 hours. HCl (4N in dioxane) was added, and the mixture was purified by RP-HPLC to give (ADC intermediate 6) as a white solid after lyophilization.

[0290] Step 3: To a solution of ADC intermediate 6 in acetonitrile / water (6 / 4, v / v) was added Mal-PEG2-Osu followed by DIEA. The reaction mixture was stirred at room temperature for 1 h and directly purified by RP-HPLC to give drug conjugate 3 as a white solid after lyophilization.

[0291] ADC Example 1 - Preparation of Trastuzumab-NMT Inhibitor ADC (DAR 5) Trastuzumab was purchased and reconstituted to give a 25 mg / mL solution. 5% v / v of 500 mM Tris, 25 mM EDTA, pH 8.5 was added to adjust the pH before reduction and conjugation. 2.5 molar equivalents of TCEP (tris(2-carboxyethyl)phosphine) relative to antibody were added from a 10 mM stock in water, and the antibody was allowed to reduce for 90 minutes. Eight molar equivalents of drug conjugate 2 were added from a 10 mM stock in DMA (dimethylacetamide), and the reduced antibody was allowed to conjugate for 60 minutes. Eight molar equivalents of NAC (N-acetylcysteine) were added from a 10 mM stock in water to quench unreacted drug conjugate 2 and allowed to react for 20 minutes. The conjugate was purified by preparative SEC (size exclusion chromatography) using a Superdex 200PG column equilibrated in PBS. Protein-containing fractions were pooled and finally filtered through a suitably sized 0.2 μm PES filter (chromatography direct / FIL-S-PES-022-13-100-S) under laminar Grade A flow. The final product was sampled for QC tests—monomer and [ADC] mg / ml by SEC-HPLC, mean DAR by PLRP, residual NMT inhibitor 1 by RP-HPLC, and endotoxin by the Endosafe dynamic colorimetric method.

[0292] ADC Example 2—Preparation of Rituximab-NMT Inhibitor ADC (DAR 5) Rituximab was purchased as a 10 mg / mL solution. 5% v / v of 500 mM Tris, 25 mM EDTA, pH 8.5 was added to adjust the pH prior to reduction and conjugation. 2.7 molar equivalents of TCEP (tris(2-carboxyethyl)phosphine) relative to antibody were added from a 10 mM stock in water, and the antibody was allowed to reduce for 120 minutes. 8 molar equivalents of drug conjugate 2 were added from a 10 mM stock in DMA (dimethylacetamide), and the reduced antibody was allowed to conjugate for 60 minutes. 8 molar equivalents of NAC (N-acetylcysteine) were added from a 10 mM stock in water to quench unreacted drug conjugate 2 and allowed to react for 20 minutes. The conjugate was purified by preparative size-exclusion chromatography (SEC) using a Superdex 200PG column equilibrated in 30 mM histidine, 0.25 M sucrose, and 100 mM NaCl. Protein-containing fractions were pooled and finally filtered through a suitably sized 0.2 μm PES filter (chromatography direct / FIL-S-PES-022-13-100-S) under grade A laminar flow. The final product was sampled for QC testing—monomer and [ADC] mg / ml by SEC-HPLC, average DAR by PLRP, residual NMT inhibitor 1 by RP-HPLC, and endotoxin by the Endosafe kinetic chromogenic assay.

[0293] ADC Example 3 was prepared using the same method as described for ADC Example 1, except that sacituzumab was used as the antibody instead of trastuzumab.

[0294] ADC Example 4 was prepared using the same method as described for ADC Example 1, except that ifinatamab was used as the antibody instead of trastuzumab.

[0295] ADC Example 5 was prepared using the same method as described for ADC Example 1, except that Drug Conjugate 3 was used instead of Drug Conjugate 2.

[0296] Preparation of ADC Example 6 ADC Example 6 was prepared using Drug Conjugate 4. Drug Conjugate 4 was prepared using the same method as described for Drug Conjugate 2, except that NMT Inhibitor 21 was used instead of NMT Inhibitor 1: [ka] (where steps 1-3 are as described for Drug Conjugate 2).

[0297] Herceptin (trastuzumab) was purchased and reconstituted to yield a 25.6 mg / mL solution. 5% v / v of 500 mM Tris, 25 mM EDTA, pH 8.5 was added to adjust the pH before reduction and conjugation. 2.55 molar equivalents of TCEP (tris(2-carboxyethyl)phosphine) relative to antibody were added from a 5 mM stock in water, and the antibody was allowed to reduce for 120 minutes. The reduced mAb was diluted 1 / 3 with PBS before conjugation. Eight molar equivalents of drug conjugate 4 were added from a 10 mM stock in DMA (dimethylacetamide), and the reduced antibody was allowed to conjugate for 90 minutes. Eight molar equivalents of NAC (N-acetylcysteine) were added from a 100 mM stock in water to quench unreacted drug conjugate 4 and allowed to react for 20 minutes. The conjugate was further buffer exchanged into PBS using G25 resin (NAP25 column), after which activated charcoal was added at a ratio of 1 mg carbon:1 mg ADC and incubated overnight at room temperature on a roller mixer at 10 rpm. The conjugate was then spun down at 4000 x G for 15 minutes to pellet the carbon, and the supernatant (ADC) was removed and filtered through a 0.2 μM PES filter. The conjugate was then further purified and concentrated by diafiltration using an Amicon 15 device, using 6x diafiltration volume (DV) of PBS, pH 7.4 for buffer exchange. The conjugate was then finally filtered through a 13 mm 0.2 μm PES filter (chromatography direct / FIL-S-PES-022-13-100-S) under grade A laminar flow and subsequently formulated to 0.02% PS80. The final product was sampled for QC tests - monomer and [ADC] mg / ml by SEC-HPLC, average DAR by PLRP, residual toxins by RP-HPLC, and endotoxin by Endosafe.

[0298] Preparation of ADC Example 7 ADC Example 7 was prepared using the same method as described for ADC Example 1, except that drug conjugate 5 was used (in this case, the linker used was GGFG). Structure of drug conjugate 5: [ka] (This can be prepared using the same method as drug conjugate 2).

[0299] Preparation of ADC Example 8 Herceptin (trastuzumab) was purchased and reconstituted to yield a 25.6 mg / mL solution. 5% v / v of 500 mM Tris, 25 mM EDTA, pH 8.5 was added to adjust the pH before reduction and conjugation. 2.55 molar equivalents of TCEP (tris(2-carboxyethyl)phosphine) relative to antibody were added from a 5 mM stock in water, and the antibody was allowed to reduce for 120 minutes. The reduced mAb was diluted 1 / 3 with PBS before conjugation. Eight molar equivalents of drug conjugate 1 were added from a 10 mM stock in DMA (dimethylacetamide), and the reduced antibody was allowed to conjugate for 90 minutes. Eight molar equivalents of NAC (N-acetylcysteine) were added from a 100 mM stock in water to quench unreacted drug conjugate 1 and allowed to react for 20 minutes. The conjugate was further buffer exchanged into PBS using G25 resin (NAP25 column), after which activated charcoal was added at a ratio of 1 mg carbon:1 mg ADC and incubated overnight at room temperature on a roller mixer at 10 rpm. The conjugate was then spun down at 4000 x G for 15 minutes to pellet the carbon, and the supernatant (ADC) was removed and filtered through a 0.2 μM PES filter. The conjugate was then further purified and concentrated by diafiltration using an Amicon 15 device, using 6x diafiltration volume (DV) of PBS, pH 7.4 for buffer exchange. The conjugate was finally filtered through a 13 mm 0.2 μm PES filter (chromatography direct / FIL-S-PES-022-13-100-S) under grade A laminar flow and then formulated to 0.02% PS80. The final product was sampled for QC tests - monomer and [ADC] mg / ml by SEC-HPLC, mean DAR by HIC, residual toxins by RP-HPLC, and endotoxin by Endosafe.

[0300] Biological Example 1: HsNMT1 and SU-DHL-10 Assays (HsNMT1 IC 50 ) IC of specific NMT inhibitors 50Values ​​were measured using a sensitive fluorescence-based assay based on the detection of CoA with 7-diethylamino-3-(4-maleimido-phenyl)-4-methylcoumarin as described in Goncalves, V. et al., Analytical Biochemistry, 2012, 421, 342-344 and Goncalves, V. et al., J. Med. Chem, 2012, 55, 3578.

[0301] (Cytotoxicity in SU-DHL-10 cell line) Certain NMT inhibitors were tested in the SU-DHL-10 cell line (human B-cell lymphoma). Compounds that demonstrate efficacy in this assay are believed to be useful as drugs for treating or preventing hyperproliferative disorders such as cancer.

[0302] Cells were seeded in 96-well microplates and treated in technical triplicates with nine increasing concentrations of compounds or cisplatin (as a positive control). IC values ​​of tested compounds and cisplatin were 50 was determined for each cell line after 72 hours of treatment.

[0303] 1. On day 1, 90 μL of various cell suspensions with cell numbers ranging from 5,000 to 8,000 cells / well were seeded into wells of a 96-well plate (Corning). The cell numbers to be seeded were determined previously. 2. All 96-well plates with cells were placed in an incubator at 37°C with 5% CO2 overnight. 3. On the second day, the cells were observed under a microscope to confirm that the cells treated with the vehicle control were in good condition. 4. A dilution series of test compounds and cisplatin was prepared at 10x the required final concentration. 10 μL / well of the 10x compound solution was added to the corresponding plate. The final volume was 100 μL / well for all plates. The final DMSO concentration was 0.1%. 5. On the fifth day (72 hours after incubation), 50 μL of CTG reagent was added to each well. 6. The contents were mixed on an orbital shaker for 5 minutes to facilitate cell lysis. 7. The plate was allowed to incubate at room temperature for 10 minutes to allow the luminescent signal to stabilize.

[0304] Luminescence was recorded using an EnVision Multi Label Reader, and data analysis was performed using GraphPad Prism 8.0.

[0305] I C 50 Concentration response curves were generated using a nonlinear regression model for a sigmoidal concentration response to calculate the IC. The formula for calculating the % of surviving cells is shown below. 50 is automatically generated by GraphPad Prism 8.0.

number

[0306] (result) The results of the above HsNMT1 and SU-DHL-10 experiments are shown in Table 2 below: Table 2: Results of Biological Example 1 [Table 6]

[0307] The results of Biological Example 1 demonstrate that the tested NMT inhibitor compounds are inhibitors of HsNMT1 and exhibit potent in vitro cytotoxic activity.

[0308] Biological Example 2: Orthotopic Breast Cancer Xenograft Model The day before tumor inoculation, NOD / SCID mice were subcutaneously implanted with an estrogen pellet (17β-estradiol, 60-day release, 0.36 mg) in the right flank. Then, on day -8, each mouse received 1 × 10 Estrogen pellets resuspended in 0.2 mL of phosphate-buffered saline (PBS) mixed with Matrigel (1:1) in the right mammary fat pad. 7 Each mouse was inoculated with live BT474 breast cancer cells. The tumor volume averaged 149.78 mm on day 0 of the study. 3 When the tumor volume reached 1000 mg / kg, the mice were assigned to a treatment group. The next day, dosing began, and all animals received intravenous administration of trastuzumab, NMT inhibitor 1, or ADC Example 1. The study was terminated on study day 35. Mice were administered vehicle alone (Group 1), 2.5 mg / kg trastuzumab (Group 2), 5 mg / kg trastuzumab (Group 3), 2.5 mg / kg ADC Example 1 (Group 4), or 5 mg / kg ADC Example 1 (Group 5) once a week for 4 weeks, or 2 mg / kg NMT inhibitor 1 was administered for 2 days followed by a 5-day rest period (Group 6). Each group consisted of 10 mice. Tumor volumes in the mice were measured 3 times a week and calculated using the formula 0.5(L×W 2 Tumor volumes were calculated using the Statistical Model (SEM). The mean tumor volumes (+SEM) for each test group at each measurement time are plotted with the last observation carried forward in Figures 1 and 2. Statistical analysis was performed on tumor readings for Groups 1, 2, 3, 4, and 5 through study day 23 (after which time point, >50% of animals in one of the test groups died; Group 2) using a two-way ANOVA, or, if values ​​were missing from a group, a mixed-effects model was fitted (PRISM GraphPad Software). Statistical analysis for Group 6 was performed through day 13 (at which point the study was terminated due to a significant loss in body weight observed). Using ROUT outlier analysis in GraphPad Prism, one mouse in Group 5 was identified as an outlier (at a 5% confidence level) across all time points and was therefore excluded from the analysis.

[0309] ΔTGI%=((mean(C)-mean(C0))-(mean(T)-mean(T0))) / (mean(C)-mean(C0))×100%, where T is the mean tumor volume of the treatment group on the day of measurement, and T0 is the mean tumor volume on day 0 of the study. C is the mean tumor volume of the control Group 1 mice on the day of measurement, and C0 is the mean tumor volume on day 0 of the study.

[0310] All animals were weighed three times a week during the study. Animals were fed Diet Gel throughout the study. The average body weights of each group during the treatment period are presented in Figures 3 and 4.

[0311] (result) The results of treatment with trastuzumab alone or ADC Example 1 on tumor size are shown in Table 3 as percent tumor growth inhibition and in Figures 1 and 2 as tumor volume (mm 3 ) Figures 1 and 2 also show tumor volumes in mice treated with NMT inhibitor 1 alone, but the study was terminated early due to significant weight loss observed in the treatment group. Table 3: Tumor growth inhibition [Table 7]

[0312] Mice treated with 2.5 mg / kg ADC Example 1 (Group 4) had significantly reduced tumor volume compared to animals treated with 2.5 mg / kg trastuzumab (Group 2; p<0.0001) (Figure 1). Furthermore, animals treated with 5.0 mg / kg ADC Example 1 (Group 5) had significantly reduced tumor volume compared to animals treated with 5 mg / kg trastuzumab (Group 3; p<0.0001) (Figure 2). Mice treated with 5.0 mg / kg ADC Example 1 (Group 5) had significantly reduced tumor volume compared to animals treated with 2.5 mg / kg ADC Example 1 (Group 4; p<0.0001) (see Table 3). Mice treated with ADC Example 1 at 5.0 mg / kg (Group 5) had significantly reduced tumor volume compared to animals treated with NMT inhibitor 1 at 2.0 mg / kg. Mice treated with NMT inhibitor 1 at 2.0 mg / kg (Group 6) had reduced tumor size (see Figures 1 and 2), but the study was stopped on Day 13 due to a significant decrease in observed body weight in this study group. ADC Example 1 administered at 2.5 mg / kg was approximately equally effective at that time compared to NMT inhibitor 1 (Group 4, see Figure 1), while ADC Example 1 administered at 5.0 mg / kg was more effective (Figure 2). The dose of NMT inhibitor 1 delivered when administered as ADC Example 1 administered at 2.5 mg / kg was approximately 100-fold less than NMT inhibitor 1 administered alone, meaning that ADC Example 1 is approximately 100-fold more potent in vivo than NMT inhibitor 1.

[0313] The effect of treatment with trastuzumab alone, NMT inhibitor 1 alone, or ADC Example 1 on mouse body weight is shown in Figures 3 and 4. Mice treated with trastuzumab or ADC Example 1 were not significantly different from vehicle controls, but mice in Group 6 (NMT inhibitor 1) had a significant decrease in body weight (Figures 3 and 4), and the study was subsequently terminated early due to a marked decrease in observed body weight.

[0314] Biological Example 3: Gastric Cancer Xenograft Model The objective of this study was to preclinically evaluate the in vivo therapeutic efficacy of an antibody-drug conjugate (ADC Example 1) in treating a subcutaneous NCI-N87 human gastric xenograft model in female BALB / c nude mice.

[0315] In this study, 143 mice were injected with 1 × 10 cells resuspended in 0.1 mL of PBS (1:1) mixed with Matrigel into the right flank for tumor development. 7 102 mice were inoculated subcutaneously with live NCI-N87 tumor cells. Tumor volumes averaged 168.08 mm on day 0 of the study. 3 When the mice reached the maximum dose, they were assigned to one of nine treatment groups. The next day, dosing began, and animals were intravenously administered vehicle control, trastuzumab, ADC Example 1, trastuzumab deruxtecan, and an isotype control antibody conjugated to NMT inhibitor 1 (isotype control). The study was terminated on study day 28. Mice were administered once weekly for two weeks with either vehicle control (Group 1), 2.5 mg / kg trastuzumab (Group 2), 5 mg / kg trastuzumab (Group 3), 2.5 mg / kg ADC Example 1 (Group 4), 5 mg / kg ADC Example 1 (Group 5), 2.5 mg / kg trastuzumab deruxtecan (Group 6), 5 mg / kg trastuzumab deruxtecan (Group 7), or 5 mg / kg isotype control antibody (Group 8). Table 4: Summary of dosing regimens [Table 8]

[0316] Tumor volumes were measured in mice three times a week, and tumor volume was calculated using the formula 0.5(L×W2). The mean tumor volume (+SEM) for each test group at each measurement is shown in Figures 5 and 6.

[0317] ΔTGI%=((mean(C)-mean(C0))-(mean(T)-mean(T0))) / (mean(C)-mean(C0))×100%, where T is the mean tumor volume of the treatment group on the day of measurement, and T0 is the mean tumor volume on day 0 of the study. C is the mean tumor volume of the control Group 1 mice on the day of measurement, and C0 is the mean tumor volume on day 0 of the study.

[0318] During the study, all animals were weighed three times a week. Animals were given Diet Gel throughout the study. The average body weights of each group during the treatment period are shown in Figures 7A (2.5 mpk study) and 7B (5 mpk study).

[0319] (result) Significant weight loss (>10%) was observed in 3 of 10 animals in Group 1 (vehicle control), 1 animal in Group 2 (2.5 mg / Kg trastuzumab), and 1 animal in Group 3 (5 mg / Kg trastuzumab); all mice regained weight by the next measurement. No significant weight loss was observed in any of the other groups.

[0320] A significant reduction (p<0.0001) in tumor volume was observed in mice treated with trastuzumab, ADC Example 1, and trastuzumab deruxtecan at all concentrations (2.5 mg / kg or 5 mg / kg) when compared to vehicle alone (Group 1). The isotype control ADC (Group 8; p=0.7935) showed no significant difference compared to vehicle alone (Group 1).

[0321] Mice treated with 5 mg / kg trastuzumab (Group 3; p<0.0001) and 5 mg / kg ADC Example 1 (Group 5; p<0.0001) showed significantly reduced tumor volume compared to mice treated with 2.5 mg / kg trastuzumab (Group 2). Mice treated with 2.5 mg / kg trastuzumab deruxtecan (Group 6; p<0.0001) and isotype control-ADC (Group 8; p<0.0001) showed significantly larger tumor volumes compared to Group 2 (2.5 mg / kg trastuzumab). There was no significant difference between ADC Example 1 at 2.5 mg / kg (Group 4; p=0.8757) and trastuzumab deruxtecan at 5 mg / kg (Group 7; p=0.9965) compared to trastuzumab at 2.5 mg / kg (Group 2).

[0322] Mice treated with 5 mg / kg ADC Example 1 (Group 5; p<0.0001) showed a significant reduction in tumor volume compared to 5 mg / kg trastuzumab (Group 3). There was no significant difference between Group 3 (5 mg / kg trastuzumab) and Group 4 (2.5 mg / kg ADC Example 1). Otherwise, all other groups (Groups 6-8) had significantly larger tumor volumes compared to Group 3 (5 mg / kg trastuzumab).

[0323] Mice treated with 5 mg / kg ADC Example 1 (Group 5; p<0.0001) showed a significant reduction in tumor volume compared to 2.5 mg / kg ADC Example 1 (Group 4). There was no significant difference between Groups 4 and 7 (5 mg / kg trastuzumab deruxtecan, p=0.9932). Otherwise, all other groups (Groups 6 and 8) had significantly larger tumor volumes compared to Group 4 (2.5 mg / kg ADC Example 1).

[0324] Mice in Group 5, treated with 5 mg / kg ADC Example 1, had significantly smaller tumor volumes compared to all other groups (p<0.0001).

[0325] Mice treated with 5 mg / kg trastuzumab deruxtecan (Group 7, p<0.0001) showed a significant reduction in tumor volume compared to 2.5 mg / kg trastuzumab deruxtecan (Group 6). Group 8 showed significantly larger tumor volumes compared to Group 6 (p<0.0001).

[0326] Mice treated with isotype control-ADC (Group 8; p<0.0001) had significantly larger tumor volumes compared to Group 7 (5 mg / kg trastuzumab deruxtecan).

[0327] Increased tumor growth inhibition (TGI) compared to the vehicle group (Group 1) was evident in all treatment groups except Group 8. Treatment with 5.0 mg / kg ADC Example 1 was the most effective when comparing tumor growth inhibition of all treatment groups (Group 5; TGI = 224.01%).

[0328] ΔTGI%=((mean(C)-mean(C0))-(mean(T)-mean(T0))) / (mean(C)-mean(C0))×100%, where T is the mean tumor volume of the treatment group on the day of measurement, and T0 is the mean tumor volume on day 0 of the study. C is the mean tumor volume of the control Group 1 mice on the day of measurement, and C0 is the mean tumor volume on day 0 of the study.

[0329] Biological Example 4: In vitro evaluation of the cytotoxicity of trastuzumab conjugated to NMT inhibitor 1 against the HER2-positive breast cancer cell line BT474 Trastuzumab was conjugated to NMT inhibitor 1 (ADC Example 1) and tested in an in vitro assay for its cytotoxic activity against the HER2-positive breast cancer cell line BT474. Cells were plated in 96-well plates at 10% confluency (8000 cells for BT474). The following day, cells were treated with 50 nM, 3.3 nM, and 0.2 nM of ADC Example 1 and controls in medium containing 250 nM Sytox Green. Plates were imaged every 4 hours for 10 days on an IncuCyte S3. Images for % phase (confluence) and green (Sytox Green) were collected and analyzed using IncuCyte software 2022A. Data were expressed as green area (μm ) normalized to phase area over time while cells were in log-phase growth. 2 ) is presented as

[0330] Figures 8-11 show that ADC Example 1 effectively induced cell killing of BT474 cells at a concentration of 0.2 nM, whereas naked trastuzumab and isotype control IgG conjugated to NMT inhibitor 1 were ineffective at killing BT474 cells under these conditions. Trastuzumab-deruxtecan also effectively killed BT474 cells, but was less effective than ADC Example 1 under these conditions. Puromycin was included as a positive cytotoxicity control.

[0331] Biological Example 5: In vitro evaluation of the cytotoxicity of trastuzumab conjugated to NMT inhibitor 1 against the HER2-negative breast cancer cell line MCF7 Trastuzumab was conjugated to NMT inhibitor 1 (ADC Example 1) and tested in an in vitro assay for its cytotoxic activity against the HER2-negative breast cancer cell line MCF7. Cells were plated in 96-well plates at 10% confluency (2000 cells for MCF7). The next day, cells were treated with 50 nM, 3.3 nM, and 0.2 nM of ADC Example 1 and controls in medium containing 250 nM Sytox Green. Plates were imaged every 4 hours for 10 days on an IncuCyte S3. Images for % phase (confluence) and green (Sytox Green) were collected and analyzed using IncuCyte software 2022A. Data were expressed as green area (μm ) normalized to phase area over time while cells were in log-phase growth. 2 ) is presented as

[0332] Figures 12-15 show that ADC Example 1, isotype control IgG conjugated to NMT inhibitor 1, trastuzumab, or trastuzumab-deruxtecan failed to induce cell killing of HER2-negative MCF7 cells at concentrations up to 50 nM under these conditions. This was expected because trastuzumab targets HER2-positive cells. Puromycin was included as a positive cytotoxicity control.

[0333] Biological Example 6: In vitro "Bystander Effect" of Trastuzumab Conjugated to NMT Inhibitor 1 HER2-negative MCF7 cells were stably transduced with pHIV eGFP (Addgene plasmid 21373) and sorted by flow cytometry to obtain a cell population that was 100% GFP-positive (green fluorescent protein). After two passages, eGFP-positive MCF7 cells were plated in a 1:1 ratio with wild-type MCF7 cells (1000 cells each) or with HER2-positive BT474 cells in 96-well plates. Cells were treated with 12.5 nM, 3.13 nM, 0.78 nM, 0.2 nM, and 0.05 nM of ADC Example 1, Reference Example 1 (trastuzumab-monomethylauristatin E (MMAE)), isotype control (defined above), trastuzumab deruxtecan, or naked trastuzumab, and controls in culture medium (50% MCF7 and 50% BT474 cell culture medium). Cells were imaged every 4 hours for 10 days on an IncuCyte S3. % Green (eGFP+MCF7) images were collected and analyzed using IncuCyte software 2022A. Data are expressed as % Green Area (μm) as a measure of eGFP+MCF7 confluence on day 9. 2 ) is presented as

[0334] Figure 16 shows that eGFP+ MCF7 cells, when cocultured with wild-type MCF7, did not suffer significant cytotoxicity when incubated with ADC Example 1 for 9 days, nor with any control or reference standard. This was expected because trastuzumab targets HER2-positive cells. However, when eGFP+ MCF7 cells were cocultured with HER2-positive BT474 cells (Figure 17), a significant loss of GFP signal was observed, corresponding to bystander killing of eGFP+ MCF7 cells. Of the three ADCs tested (ADC Example 1, Reference Example 1, and trastuzumab deruxtecan), ADC Example 1 was the most effective at inducing bystander killing, ADC Reference Example 1 was second most effective, and trastuzumab deruxtecan was the least effective. Naked trastuzumab and isotype control IgG conjugated to NMT inhibitor 1 were used as controls, and neither induced bystander killing.

[0335] Biological Example 7: In vitro evaluation of cytotoxicity of antibodies conjugated to NMT inhibitor 1 NMT inhibitor 1 was conjugated to trastuzumab (ADC Example 1) and sacituzumab (ADC Example 3) as described. NMT inhibitor 26 was conjugated to trastuzumab (ADC Example 5). ADC Examples 1, 3, and 5 were tested for their cytotoxic activity against a panel of cell lines including BT474, JIMT 1, NCI N87, NCI H292, IM95-m, ZR-75-30, and NCI H2170 (Figures 18-24).

[0336] NMT inhibitor 1 was conjugated to ifinatamab (ADC Example 4) as described and tested for its cytotoxic activity against a panel of prostate cancer cell lines, including LNCaP, C42, and VCaP (Figures 25-27). Cells were plated at a defined initial density in 96-well flat-bottom plates in the appropriate growth medium. After 24 hours, dosing solutions were added with serial dilutions of each test compound spanning a concentration range of either 10 nM to 0.001 nM for NMT inhibitor 1 or 50 nM to 0.005 nM for various ADCs. Cell proliferation was measured by the CellTiter-Glo 2.0 solution cell viability assay after 144 hours of exposure. Percent inhibition of viability at the endpoint (after blank correction, time zero correction, and logarithmic transformation of compound concentrations) relative to the DMSO vehicle control was calculated using the following equation: % survival = (day 6 luminescence of tested cells - blank) - (day 0 luminescence - blank) / (average of day 6 luminescence of vehicle-treated cells - blank) - (day 0 luminescence - blank)

[0337] (result) Table 5 shows the inhibition of cell viability, quantified as above and defined as the concentration of inhibitor that reduces the absolute IC50 response by half or the concentration required to reduce the relative IC50 curve to the midpoint between the upper and lower plateaus of the curve. The complete concentration-response curves for each cell line are shown in Figures 18-27 (mean + / - SEM). Table 5: Inhibition of cell viability [Table 9]

[0338] Biological Example 8: Evaluation of the effect of test articles as single agents on cell viability of gastric cancer organoid lines using the CellTiter-Glo (CTG) luminescent cell viability assay (method) Day -1: Shear patient-derived gastric cancer organoids (PDXO) to a uniform size and combine the required number of organoids 1:1 with 50% Matrigel to produce organoids of appropriate size for performing screening.

[0339] Day 0: Organoid seeding 1. Harvest organoids from each well of a 6-well plate containing 2 ml of organoid medium by adding 20 μl of 100× dispase solution to each well. 2. Return the plate to the 37°C incubator for 30 minutes. 3. Collect organoids from all wells and pipette them through a pre-wetted 100 μm filter into a 50 ml plastic tube. 4. Once all wells have been filtered through the 100 μm filter, filter the flow-through through a pre-wetted 20 μm filter. 5. Invert the 20 μm filter and allow the organoids to recover in a new 50 ml tube. 6. Collect and resuspend the organoids in the corresponding culture medium. Count the organoids to obtain the concentration. 7. Adjust the cell concentration to the appropriate concentration with culture medium. 8. Add Matrigel to a final concentration of 5% v / v and keep the organoid suspension on ice. 9. Add 40 μL of cell suspension with a Multidrop dispenser to a 384-well plate with the corresponding culture medium at the following seeding densities: [Table 10] 10. Place screening plates back into the incubator before adding compound and taking the day 0 read. Two duplicate plates were prepared: one for the day 0 read (TO) and one for the endpoint read.

[0340] Day 0: Compound treatment and baseline CTG (cell titer glo) reading on day 0 11. Add test articles according to the drug dilution scheme and plate map on the Tecan D300e. 12. Return the screening plate to the incubator. 13. For the day 0 reading plate, add 40 μL of CTG 3D per well and read the luminescent signal on an Envision plate reader.

[0341] Day 6: End-point CTG reading 1. Read the assay plate for luminescent CTG signal at the end of the assay: add 40 μL of CTG 3D per well with the Multidrop dispenser, mix the contents on a plate shaker for 5 minutes, and incubate the plate in the dark at room temperature for 30 minutes. Read the luminescent signal on the Envision plate reader.

[0342] Data were graphed using GraphPad Prism. To calculate absolute IC50s, concentration-response curves were fitted using a nonlinear regression model with a sigmoidal dose-response with variable slope. The formula for calculating survival rates is shown below. Absolute IC50s were calculated according to the dose-response curves generated by GraphPad Prism. Viability (%) = (Lum test article - Lum medium control) / (Lum vehicle control - Lum medium control) x 100%.

[0343] Gastric cancer organoids were incubated with a range of concentrations of (1) isotype control ADC, (2) ADC Example 1, (3) staurosporine as positive control, (4) trastuzumab, or (5) trastuzumab-deruxtecan (trastuzumab-DXd) for 5 or 6 days.Table 6 shows IC50 (μM), maximum inhibition (%) and area under the curve. Table 6: Results of organoid studies [Table 11] TIFF2025530213000110.tif247170TIFF2025530213000111.tif247170TIFF2025530213000112.tif40170

[0344] Concentration response curves of 16 gastric cancer organoids are shown in Figures 28 to 43. Data show organoid survival (mean + / - SEM) after treatment with isotype control, ADC Example 1, trastuzumab-deruxtecan (DXd), and trastuzumab conjugated to NMT inhibitor 1.

[0345] These results show that in the majority of gastric cancer organoids, ADC Example 1 is the most effective, as shown by its IC50 value, compared with control.In certain organoids, ADC Example 1 has the same level of activity as trastuzumab-DXd, as shown by IC50 value.

[0346] Biological Example 9: Maximum Tolerated Dose Study of ADC Example 1 After a Single Intravenous Injection in Male Cynomolgus Monkeys Six previously used male cynomolgus monkeys were used in this MTD study. The service provider laboratory conducting this study held certification from the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC). The study's animal care and use application was submitted to the service provider's Institutional Animal Care and Use Committee (IACUC) for approval. The IACUC reviewed the protocol for consistency with TSP IACUC policies and procedures. The monkeys were quarantined at the vendor for at least two weeks before shipping to the service provider's facility. Animals were screened for active infections with tuberculosis, herpes B, simian acquired immunodeficiency virus (SIV), simian retrovirus (SRV), simian T-cell lymphotropic virus (STLV), Shigella flexneri, and Salmonella typhimurium, and were evaluated for normal hematology and clinical chemistry parameters. Evidence of parasites was determined by serology or fecal analysis.

[0347] Animals were randomized into three groups of two animals per group and administered the test article, ADC Example 1, by slow intravenous infusion (5 minutes) according to Table 7. Table 7: Dosing regimen for Biological Example 9 [Table 12]

[0348] Animals were assessed at life stages according to Table 8. Table 8: Evaluation Protocol for Biological Example 9 [Table 13]

[0349] Blood samples were collected from each test animal on the following days: the day before dosing (Day 1), study Day 2, and at the time of termination (Day 7 or Day 11). Animals were fasted overnight before sample collection for clinical pathology. Blood samples were collected by peripheral vein puncture. Blood samples were processed and analyzed for hematology, clinical biochemistry, and coagulation. Each of the 5, 10, and 20 mg / kg doses was well tolerated by the respective groups, and no significant changes were observed in hematological and clinical chemistry parameters. In particular, the 5 and 10 mg / kg groups showed no changes. Figures 44a and 44b show representative data for the 20 mg / kg group. Specifically, mean (+ / - SEM) levels of hematological markers were plotted as normalized to baseline (pre-dose) values. No changes were observed on Day 7, but relatively small, but not significant, changes were observed on Day 11.

[0350] Biological Example 10: In vivo tolerability study of ADC Example 1 in non-tumor-bearing female CD1 mice The objective of this study was to evaluate the in vivo tolerability of ADC Example 1 in female CD1 mice. Body weights were measured daily, and terminal whole blood and serum were collected from all animals at the time of sacrifice for complete blood counts and blood chemistries.

[0351] Twenty-four mice were randomly assigned to four groups, and dosing began the following day. All animals received a single intravenous dose of either PBS (Group 1: vehicle control) or ADC Example 1 (Groups 2, 3, and 4; 25, 50, or 100 mg / kg). The study was terminated on Day 7. Daily food intake was measured, and complete necropsies were performed on all animals and two untreated animals at the end of the study.

[0352] Significant weight loss (>10%) was observed for three animals in Group 4 (100 mg / kg ADC Example 1) on study day 5, and Group 4 was terminated on day 6 (Figure 45). No significant weight loss was observed in any of the other groups (Figure 45), and no clinical findings were observed for any of the animals being tested.

[0353] Whole blood for complete blood counts (CBC) and serum for blood chemistry were collected from all animals being studied at the time of slaughter. Figure 46 shows the mean values ​​of AST (aspartate transaminase), ALT (alanine transaminase), ALP (alkaline phosphatase), LDH (lactate dehydrogenase), CK (creatine kinase), and GGT (gamma-glutamyltransferase) in the blood of animals at the time of slaughter. Significant changes were observed only in Group 4 (100 mg / Kg). Figure 47 shows the mean values ​​of cells: LYM (lymphocytes), MON (monocytes), NEU (neutrophils), RBC (red blood cells), HGB (hemoglobin), and PLT (platelets) in the blood of animals at the time of slaughter. Significant changes were observed only in Group 4 (100 mg / Kg ADC Example 1).

[0354] In conclusion, ADC Example 1 is well tolerated in mice after a single IV dose of up to 50 mg / Kg, 10-fold higher than the effective dose observed for Biological Examples 2 and 3.

[0355] Biological Example 11: LNCaP Prostate Cancer Xenograft Model The purpose of this study was to evaluate the efficacy of ADC Example 4 in male NOD SCID mice bearing LNCaP tumors.

[0356] A total of 84 male NOD SCID mice, 5-8 weeks old and weighing 25-30 g, were used in the study. 1 × 10 mice were cultured at 78% survival rate and approximately 70-80% confluency. 7 LNCaP tumor cells were implanted subcutaneously into the flanks of male NOD SCID mice. Tumors grew to approximately 80–100 mm. 3 When tumor volume reached 1000 mg / kg, the animals were assigned to treatment groups shown in Table 9 below, with 10 mice per group assigned to each group with similar tumor volume means and distributions. Mice were treated with vehicle only, unconjugated ifinatamab, ifinatamab-deruxtecan (ifinatamab-DXd), or ADC Example 4. Table 9: Dosing regimen for Biological Example 11 [Table 14] Observation period: 35 days Dosage volume: 5 mL / kg for all IV doses

[0357] No adverse responses were observed to any dose during the course of the study, and the mean body weights of each group remained within 10% of pre-treatment levels (Figure 49).

[0358] Individual cases of weight loss >10% were observed at various time points throughout the study. Three weeks after the first animals entered treatment, all mice were offered DietGel to ameliorate weight loss. No animals were euthanized early due to weight loss, and these instances of weight loss were likely related to tumor burden.

[0359] Animals receiving three Q7D doses of 10 mg / kg ADC Example 4 exhibited significantly higher mean body weights than animals receiving vehicle control treatment on Study Day 28 (Dunnett's one-way ANOVA, p=0.0046). No other treatment groups were significantly different from vehicle in terms of body weight at this time point.

[0360] For welfare reasons, two animals were euthanized early. One was euthanized on day 26 of treatment with 10 mg / kg ifinatamab, while the second was euthanized on day 33 of treatment with 5 mg / kg ifinatamab-DXd. Both animals were euthanized primarily due to gasping for breath. At necropsy for each, large spontaneous thymic tumors were noted.

[0361] Tumors in the vehicle-treated group grew steadily over the course of the study, reaching a size of 752 ± 89.4 mm by study day 28. 3 reached an average volume of

[0362] Treatment with either 10 mg / kg or 5 mg / kg ifinatamab had no significant effect on LNCaP tumor volume at day 28, and animals receiving this treatment exhibited tumor growth curves similar to those of animals treated with vehicle alone (Figure 48, Table 10).

[0363] Treatment with 10 mg / kg ifinatamab-DXd significantly reduced the mean volume of LNCaP tumors by day 28 compared to vehicle controls. Animals receiving this treatment mostly exhibited a slower tumor growth rate than control animals. A dosage of 5 mg / kg ifinatamab-DXd did not significantly slow LNCaP tumor growth (Figure 48, Table 10).

[0364] All animals receiving 10 mg / kg ADC Example 4 showed tumor regression within 3 weeks of initiating treatment (Figure 48, Table 10), and this therapy resulted in a significant reduction in mean tumor volume (TV) compared to vehicle (Mann-Whitney) from day 7 onwards. By day 28, each tumor had regressed to ≤25% of its volume at the start of treatment.

[0365] Similarly, animals receiving 5 mg / kg of ADC Example 4 showed a significant reduction in tumor volume (Mann-Whitney) compared to the control group from day 7 to day 28. By day 28, all but one animal exhibited tumor volumes lower than those recorded at the start of treatment. Table 10. Tumor volume comparison of treatment groups. Adjusted p-value calculated by Kruskal-Wallis test and Dunn's multiple comparisons against vehicle control. TGI% = ((mean (C) - mean (C0)) - (mean (T) - mean (T0))) / (mean (C) - mean (C0)) × 100%, where T is the mean tumor volume of the treatment group on the day of measurement, and T0 is the mean tumor volume on day 0 of the study. C is the mean tumor volume of mice in the control group 1 on the day of measurement, and C0 is the mean tumor volume on day 0 of the study. [Table 15]

[0366] Biological Example 12: VCaP Prostate Cancer Xenograft Model The objective of this study was to preclinically evaluate the in vivo therapeutic efficacy of ADC Example 4 in treating a subcutaneous VCaP human prostate cancer xenograft model in intact male CB17 / SCID mice.

[0367] In this study, 144 mice were injected into the right anterior flank with 1 × 10 mice resuspended in 0.1 mL of PBS (1:1) mixed with Matrigel for tumor development. 7 Live VCaP tumor cells were inoculated subcutaneously on day -20 of the study. The tumor volume averaged approximately 162.16 mm on day 0 of the study. 3 When the study population reached 100, 80 mice were assigned to eight treatment groups. The following day, dosing began, and all animals received intravenous administration of ADC Example 4, ifinatamab-deruxtecan (ifinatamab-Dxd), or unconjugated ifinatamab. All mice received two doses of study drug, on Study Day 1 and Study Day 8. The study ended on Study Day 30.

[0368] The eight groups were assigned as follows: Group 1: Vehicle control Arm 2: 5mpk unconjugated ifinatamab Group 3: 2.5mpk unconjugated ifinatamab Group 4: 5mpk ifinatamab-deruxtecan Group 5: 2.5mpk ifinatamab-deruxtecan Group 6 10mpk ADC Example 4 Group 7 5mpk ADC Example 4 Group 8 2.5mpk ADC Example 4

[0369] No significant weight loss was observed in any of the animals tested (Figure 51).

[0370] A significant reduction (p<0.0001) in tumor volume was observed in mice treated with all concentrations of ADC Example 4 (Group 8, 2.5 mg / kg; Group 7, 5 mg / kg; and Group 6, 10 mg / kg) compared to vehicle alone (Group 1). Ifinatamab-Dxd at 5 mg / kg (Group 4; p=0.0078) exhibited significantly higher tumor volumes compared to vehicle alone (Group 1), whereas ifinatamab-Dxd at 2.5 mg / kg (Group 5; p=0.8127) and both concentrations of unconjugated ifinatamab (Group 2; p=0.1104 and Group 3; p=0.6703) did not exhibit significant differences compared to vehicle alone (Group 1). See Figure 50.

[0371] Mice treated with all three concentrations of ADC Example 4 (Group 6, 10 mg / kg; Group 7, 5 mg / kg, and Group 8, 2.5 mg / kg) showed significant reductions in tumor volume compared to all other treatment groups (Groups 2-5; p<0.0001). There was also evidence of a dose response, with the greatest tumor volume reductions in Group 6 (10 mg / kg), followed by Group 7 (5 mg / kg), and then Group 8 (2.5 mg / kg); significant differences were observed between each group (p<0.0001). See Figure 50.

[0372] No significant differences in tumor volume were observed between mice treated with 2.5 mg / kg ifinatamab-Dxd (Group 5) and mice treated with Groups 2, 3, and 4 (5 mg / kg unconjugated ifinatamab, 2.5 mg / kg unconjugated ifinatamab, and 5 mg / kg ifinatamab-Dxd, respectively). Mice treated with 5 mg / kg ifinatamab-Dxd (Group 4) had significantly larger tumor volumes compared to mice treated with 5 mg / kg unconjugated ifinatamab (Group 2, p=0.0002) and 2.5 mg / kg unconjugated ifinatamab (Group 3, p=0.0216). See Figure 50.

[0373] An increase in tumor growth inhibition (ΔTGI) compared to the vehicle group (Group 1) was evident in all treatment groups. When comparing tumor growth inhibition across all treatment groups, treatment with 10 mg / kg and 5 mg / kg ADC Example 4 was most effective (Group 6; ΔTGI = 114.03% and Group 7; ΔTGI = 114.69%). See Table 11. Table 11. Dosing regimen and results for Biological Example 12 [Table 16]

[0374] Biological Example 13: JIMT-1 Breast Cancer Xenograft Model The objective of this study was to preclinically evaluate the in vivo therapeutic efficacy of ADC Example 3 in treating a subcutaneous JIMT-1 human breast xenograft model in female NOD / SCID mice.

[0375] In this study, 128 mice were injected with 5 × 10 , resuspended in 0.1 mL of PBS, into the right anterior flank for tumor development. 6 Live JIMT-1 tumor cells were subcutaneously inoculated on day 15 of the study. The tumor volume was approximately 160.66 mm on average on day 0 of the study. 3 When the mice reached 60 days old, 80 mice were assigned to eight treatment groups. The next day, dosing began and all animals received ADC Example 3, sacituzumab govitecan, or unconjugated sacituzumab intravenously. The study ended on Study Day 60. The eight groups were assigned as follows: Group 1: Vehicle control Arm 2: 5mpk unconjugated sacituzumab Arm 3: 2.5mpk unconjugated sacituzumab Group 4 5 mg / kg sacituzumab govitecan (ADC Example 3, 5 mg / kg added on days 27 and 34) Group 5: 2.5 mg / kg sacituzumab govitecan (ADC Example 3, 5 mg / kg added on day 27) Group 6 10mpk ADC Example 3 Group 7 5mpk ADC Example 3 Group 8 2.5mpk ADC Example 3

[0376] Significant weight loss (>10%) was observed in one animal in Group 7. No significant weight loss was observed in any of the other groups tested (see Figures 54 and 55). On day 10 of the study, one animal in Group 2 was found dead.

[0377] When compared with vehicle alone (Group 1), mice treated with all concentrations of ADC Example 3 (Group 8, 2.5 mg / kg; Group 7, 5 mg / kg, and Group 6, 10 mg / kg) showed significant reductions in tumor volume (p<0.0001). Both concentrations of sacituzumab govitecan (Group 4, 5 mg / kg, p<0.0001; Group 5, 2.5 mg / kg, p=0.0348) showed significant reductions in tumor volume compared with vehicle alone (Group 1). When compared with vehicle alone (Group 1), unconjugated sacituzumab in Group 2 (5 mg / kg, p=0.0028) showed significant reductions in tumor volume, but no significant difference was seen in Group 3 (2.5 mg / kg, p=0.0586). Data for the 5 mg / kg and 10 mg / kg groups are shown in Figure 52. Data for the 2.5 mg / kg group are shown in Figure 53.

[0378] Mice treated with all three concentrations of ADC Example 3 (Group 6, 10 mg / kg; Group 7, 5 mg / kg, and Group 8, 2.5 mg / kg) showed significant reductions in tumor volume compared to all other treatment groups (Groups 2-5; p<0.0001). There was also evidence of a dose response, with Groups 6 (10 mg / kg) and 7 (5 mg / kg) showing the greatest tumor volume reductions (p<0.0001) compared to Group 8 (2.5 mg / kg).

[0379] Sacituzumab govitecan at 5 mg / kg in Group 4 demonstrated significant reductions in tumor volume compared with unconjugated sacituzumab in Groups 2 (5 mg / kg, p = 0.0097) and 3 (2.5 mg / kg, p = 0.0499), but no significant differences were observed between Group 5 (2.5 mg / kg sacituzumab govitecan) and Groups 2 (p = 0.8424) or 3 (p = 0.9995). There was also no significant difference between the two concentrations of unconjugated sacituzumab in Groups 2 (5 mg / kg) and 3 (2.5 mg / kg) (p = 0.9837). However, there was a significant difference between the two concentrations of sacituzumab govitecan in Groups 4 (5 mg / kg) and 5 (2.5 mg / kg) (p = 0.3802).

[0380] An increase in tumor growth inhibition (ΔTGI) compared to the vehicle group (Group 1) was evident in all treatment groups. When comparing tumor growth inhibition across all treatment groups, treatment with 10 mg / kg and 5 mg / kg ADC Example 3 was most effective (Group 6: ΔTGI=121.55% and Group 7: ΔTGI=122.04%).

[0381] Because treatment with sacituzumab govitecan was only partially effective in Group 4, additional doses of ADC Example 3 were administered IV at 5 mpk on Study Days 27 and 34. This resulted in a significant reduction in tumor volume compared to vehicle controls (see Figure 52). Group 5 also received ADC Example 3 (5 mpk IV) on Study Day 27 (see Figure 53), but no significant response was observed. Table 12. Dosing regimens and results for Biological Example 13 [Table 17]

[0382] Biological Example 14: In vitro evaluation of the cytotoxicity of trastuzumab conjugated to NMT inhibitor 1 against the HER2-positive breast cancer cell line BT474 Trastuzumab was conjugated to NMT inhibitor 1 (ADC Example 8) and tested in an in vitro assay for its cytotoxic activity against the HER2-positive breast cancer cell line BT474. Cells were plated in 96-well plates at 10% confluency (8000 cells for BT474). The next day, cells were treated with 50 nM, 3.13 nM, and 0.2 nM of ADC Example 8, as well as controls, in medium containing 250 nM Sytox Green. Plates were imaged every 4 hours for 10 days on an IncuCyte S3. Images for % phase (confluence) and green (Sytox Green) were collected and analyzed using IncuCyte software 2022A. Data were expressed as green area (μm ) normalized to phase area over time while cells were in logarithmic phase growth. 2 ) is presented as

[0383] Figure 56 shows that ADC Example 8 effectively induced cell killing of BT474 cells at a concentration of 0.2 nM, whereas naked trastuzumab and isotype control IgG conjugated to NMT inhibitor 1 were ineffective at killing BT474 cells under these conditions (see Figures 9 and 11). Trastuzumab-deruxtecan also effectively killed BT474 cells, but was less effective than ADC Example 8 under these conditions (see Figure 10). Puromycin was included as a positive cytotoxicity control.

[0384] Biological Example 15: In vitro evaluation of the cytotoxicity of trastuzumab conjugated to NMT inhibitor 1 against the HER2-negative breast cancer cell line MCF7 Trastuzumab was conjugated to NMT inhibitor 1 (ADC Example 8) and tested in an in vitro assay for its cytotoxic activity against the HER2-negative breast cancer cell line MCF7. Cells were plated in 96-well plates at 10% confluency (2000 cells for MCF7). The next day, cells were treated with 50 nM, 3.13 nM, and 0.2 nM of ADC Example 8, as well as controls, in medium containing 250 nM Sytox Green. Plates were imaged every 4 hours for 10 days on an IncuCyte S3. Images for % phase (confluence) and green (Sytox Green) were collected and analyzed using IncuCyte software 2022A. Data were expressed as green area (μm ) normalized to phase area over time while cells were in log-phase growth. 2 ) is presented as

[0385] Figures 13, 14, 15, and 57 show that ADC Example 8, isotype control IgG conjugated to NMT inhibitor 1, trastuzumab, or trastuzumab-deruxtecan failed to induce cell killing of HER2-negative MCF7 cells at concentrations up to 50 nM under these conditions. This was expected because trastuzumab targets HER2-positive cells. Puromycin was included as a positive cytotoxicity control.

[0386] (Conclusion) The results of Biological Example 1 demonstrate that the tested NMT inhibitor compounds are inhibitors of HsNMT1 and exhibit potent in vitro cytotoxic activity.

[0387] The results of Biological Example 2 demonstrate that the tested ADCs of the invention exhibit potent in vivo cytotoxic activity and have improved in vivo tolerability when compared to NMT inhibitor 1.

[0388] The results of Biological Example 3 show that the ADC of the invention tested, ADC Example 1, exhibits improved efficacy compared to other treatment groups (see Table 4) in inhibiting tumor growth in a gastric cancer xenograft mouse model, with no significant weight loss observed.

[0389] The results of Biological Example 4 demonstrate that the ADC of the invention tested, ADC Example 1, effectively induces cell killing of the HER2-positive breast cancer cell line BT474. The ADC of the invention was more effective at inducing cell killing compared to the other compounds tested: trastuzumab, trastuzumab deruxtecan, and an isotype control.

[0390] The results of Biological Example 6 show that ADC Example 1 was most effective in inducing bystander killing in eGFP+ MCF7 cells co-cultured with HER2-positive BT474 cells compared to other ADCs and controls.

[0391] The results of Biological Example 7 demonstrate that various ADCs of the invention (specifically, ADC Examples 1 and 3-5) have cytotoxic activity against various cancer cell lines as measured in cell viability assays. Various ADCs of the invention (specifically, ADC Examples 1 and 3-5) had improved cytotoxicity compared to their respective controls, as shown by the data in Figures 18-27.

[0392] The results of Biological Example 8 show that in most gastric cancer organoids, ADC Example 1 is the most effective, as shown by its IC50 value compared with control.In certain organoids, ADC Example 1 has the same activity as trastuzumab-DXd, as shown by IC50 value.

[0393] The results of Biological Example 9 show that ADC Example 1 was well tolerated in a monkey model, even at a maximum dose of 20 mg / kg / day, with no significant changes in hematological or clinical chemistry parameters.

[0394] The results of Biological Example 10 show that ADC Example 1 is well tolerated in mice following a single IV dose of up to 50 mg / Kg, 10-fold higher than the effective dose observed in Biological Examples 2 and 3.

[0395] The results of Biological Example 11 show that ADC Example 4 was well tolerated in mice at both 5 mg / kg and 10 mg / kg doses in the LNCaP prostate cancer xenograft model. Animals receiving these doses of ADC Example 4 showed significant reductions in tumor volume, unlike treatment with ifinatamab alone, which had no significant effect on tumor volume. ADC Example 4 also performed better than ifinatamab-DXd at both doses.

[0396] The results of Biological Example 12 show that ADC Example 4 was well tolerated in mice at doses of 2.5 mg / kg, 5 mg / kg, and 10 mg / kg in a VCaP human prostate cancer xenograft model. ADC Example 4 reduced tumor volume to a greater extent than infinatamab-Dxd and unconjugated infinatamab. In fact, the use of unconjugated infinatamab showed no significant difference compared to vehicle alone. ADC Example 4 produced the greatest increase in tumor growth inhibition compared to the control (see Table 11).

[0397] The results of Biological Example 13 show that ADC Example 3 was well tolerated in mice at doses of 2.5 mg / kg, 5 mg / kg, and 10 mg / kg in the JIMT-1 human breast xenograft model. Mice treated with all three concentrations of ADC Example 3 showed significant reductions in tumor volume compared to all other treatment groups, as also shown by the ΔTGI results in Table 12, where ADC Example 3 resulted in the greatest tumor growth inhibition.

[0398] The results of Biological Example 14 demonstrate that the ADC of the invention tested, ADC Example 8, effectively induces cell killing of the HER2-positive breast cancer cell line BT474. The ADC of the invention was more effective at inducing cell killing compared to the other compounds tested: trastuzumab, trastuzumab deruxtecan, and an isotype control.

[0399] Overall, therefore, the ADCs of the invention are believed to be useful as pharmaceuticals, particularly in the treatment of hyperproliferative disorders such as cancer.

[0400] Throughout this specification and the claims that follow, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" will be understood to imply the inclusion of a specified integer, step, group of integers, or group of steps, but not the exclusion of any other integer, step, group of integers, or group of steps.

[0401] All patents and patent applications referred to herein are incorporated by reference in their entirety.

Claims

1. An antibody drug conjugate (ADC), or a salt thereof, comprising an NMT inhibitor conjugated to an antibody via a linker.

2. A salt of the ADC of claim 1.

3. 3. The salt according to claim 1 or 2, which is a pharmaceutically acceptable salt.

4. The ADC of claim 1.

5. The ADC or salt thereof according to any one of claims 1 to 4, wherein the NMT inhibitor is a compound of formula (I): 【Chemical 1】 (In the formula: Y is -CH-, -C(R 2 )-, and -N-; R 1 is a group of formula -XLA; X represents -O-; L is -(CH 2 ) m - represents; m is 1, 2, or 3; A is a 6- to 10-membered aromatic carbocyclic ring or a 5- to 10-membered aromatic heterocyclic ring, and the aromatic carbocyclic ring or heterocyclic ring is selected from the group consisting of -F, -Cl, -Br, -OCH 3 , -OCF 3 , -CN, up to three halogens, hydroxyl, or -OC 1-4 -C optionally substituted with alkyl groups 1-6 Alkyl, -S(O)C 1-4 Alkyl, -S(O) 2 C 1-4 Alkyl, -C(O)N(R 9 ) 2 , -C(O)N(R 13 )C 1-4 Alkyl OC 1-4 Alkyl, -C(O)N(C 1-4 Alkyl OC 1-4 alkyl) 2 , -CH 2 C(O)N(R 9 ) 2 , -CH 2 C(O)N(R 13 )C 1-4 Alkyl OC 1-4 Alkyl, -CH 2 C(O)N(C 1-4 Alkyl OC 1-4 alkyl) 2 , -S(O) 2 NHC 1-4 Alkyl, -S(O) 2 N(C 1-4 alkyl) 2 , -NHC 1-4 Alkyl, -N(C 1-4 alkyl) 2 , -NHC(O)C 1-4 Alkyl, -NHC(O)CF 3 , -NHS(O) 2 C 1-4 Alkyl, CH 2 N(R 13 ) 2 , C.H. 2 N(R 13 )C(O)C 1-4 Alkyl, CH 2 N(R 13 )S(O) 2 C 1-4 Alkyl, -CH 2 S(O) 2 C 1-4 Alkyl, and CO 2 optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of H; s is 0, 1, 2, or 3; Each R 2 -F, -Cl, -Br, -OCH 3 , -OCF 3 , -CN, -C optionally substituted by up to three halogen or hydroxyl groups 1-4 Alkyl, -S(O)C 1-4 Alkyl, -S(O) 2 C 1-4 Alkyl, -S(O) 2 NHC 1-4 Alkyl, -S(O) 2 N(C 1-4 alkyl) 2 , -NHC 1-4 Alkyl, -N(C 1-4 alkyl) 2 , -NHC(O)C 1-4 Alkyl, -NHC(O)CF 3 , and -NHS(O) 2 C 1-4 independently selected from the group consisting of alkyl; q is 0 or 1; R 3 is hydrogen or methyl; R 4 is hydrogen or methyl; R 5 is hydrogen; R 6 is hydrogen or up to three of -F, -Cl, -Br, -OH, -OCH 3 , -OCF 3 or C optionally substituted with a -CN group 1-6 is alkyl; If present, R 10 is hydrogen or methyl; If present, R 11 is hydrogen or methyl; or R 3 Groups and R 5 The group and the intervening atom may be the intervening atom and a bond, or the intervening atom and -(CHR a ) r -, or R 10 group and the R 5 The group and the intervening atom are separated by a -(CHR a ) r -, forming a 3- to 7-membered non-aromatic heterocycle consisting of; r is 1, 2, 3, 4, or 5; R a is hydrogen or methyl; Each R 7 is hydrogen, halogen, C 1-4 C optionally substituted with alkoxy and one, two, or three halogens 1-4 independently selected from the group consisting of alkyl; and R 8 is hydrogen and C 1-4 alkyl; Each R 9 is hydrogen and C 1-4 alkyl, or two R 9 the groups and the N to which they are attached form a 4- to 7-membered non-aromatic heterocycle, which optionally contains one or two additional heteroatoms selected from N, O, and S; Each R 13 is hydrogen and C 1-4 alkyl; and wherein: i) E, J, and G are each C(R 7 ), K is carbon, and Q is N(R 8 ) and M is nitrogen; ii) E, J, and G are each C(R 7 ) and K, Q, and M are each nitrogen; or iii) E, J, G, and M are each C(R 7 ) and K and Q are each nitrogen.

6. 6. The ADC of claim 5, wherein the NMT inhibitor is a compound of formula (IA^^): or a salt thereof. 【Chemistry 2】 (In the formula: R 1 is a group of formula -XLA; A is 4-pyrazolyl, the pyrazolyl being methyl and —C(O)N(CH 3 ) 2 optionally substituted with up to three substituents selected from: X is -O-; L is -(CH 2 ) m - and; m is 2; R 2' is selected from the group consisting of fluorine or chlorine (preferably fluorine); R 2'' is selected from the group consisting of hydrogen, fluorine, or chlorine; q is 0; R 3 is hydrogen or methyl; R 4 is hydrogen or methyl; R 5 is hydrogen; R 6 is hydrogen or methyl; or R 3 Groups and R 6 the group and the intervening atom(s) form a 3- to 7-membered non-aromatic heterocyclic ring consisting of the intervening atom(s) and the bond; E, J, G, K, Q, and M are: i) E, J, and G are each CH, K is carbon, and Q is N(R 8 ), M is nitrogen; and R 8 is hydrogen or methyl; or ii) E, J, G, and M are each CH and K and Q are each nitrogen; However, A is 1 or less -C(O)N(CH 3 ) 2 (substituted with a group).

7. The ADC or salt thereof according to any one of claims 1 to 6, wherein the NMT inhibitor is a compound of formula (II): 【Chemistry 3】 (In the formula: R 1 is H or -CH 3 and R 2 is H or F).

8. The NMT inhibitor is 4-(2-{2-[3-(2-aminoethyl)imidazo[1,2-a]pyridyl-6-yl]-5-chlorophenoxy}ethyl)-N,N,1,5-tetramethyl-1H-pyrazole-3-carboxamide: 【Chemistry 4】 or a salt thereof.

9. The ADC or salt thereof according to any one of claims 1 to 4, wherein the NMT inhibitor is a compound of formula (III) or (IV): 【Chemistry 5】 (In the formula: n 1 is 0, 1, 2, 3, 4, 5, or 6; Ring A * is an optionally substituted nitrogen-containing aryl group, where ring A * Each substitutable carbon or nitrogen in 5A and wherein ring A is optionally and independently substituted by * contains an -NH- moiety, the nitrogen is C 1-6 optionally substituted with alkyl (e.g., methyl); and wherein R 4A and Ring A * may together with the atoms to which they are attached form a cyclic group; Ring B * is an optionally substituted aryl or heteroaryl group, wherein ring B * Each substitutable carbon or heteroatom in 3A optionally and independently substituted by; One of W and X may be absent. 11A , R 11A optionally substituted hydrocarbyl (e.g., C 1-8 alkyl, alkenyl, alkynyl, or haloalkyl), and R 12A optionally substituted with -(CH 2 ) k1 -heterocyclyl; k 1 is 0, 1, 2, 3, 4, 5, or 6; R 1A is hydrogen; R 2A , R 3A , R 4A , and R 5A is hydrogen, R 12A , R 12A optionally substituted hydrocarbyl (e.g., C 1-6 alkyl, alkenyl, alkynyl, or haloalkyl), and one or more R 12A optionally substituted with -(CH 2 ) L1 -heterocyclyl; where R 2A together with W or X, one or more R 12A may form an optionally substituted heterocycle; and wherein R 3A and R 5A One or more of, together with the atom to which they are attached, R 12A may form an optionally substituted carbocycle, e.g., heterocyclyl; L 1 is 0, 1, 2, 3, 4, 5, or 6; where: Each R 11A and R 12A is halogen, trifluoromethyl, cyano, thio, nitro, oxo, =NR 13A , -OR 13A , -SR 13A , -C(O)R 13A , -C(O)OR 13A , -OC(O)R 13A , -NR 13A COR 14A , -NR 13A CON(R 13A ) 2 , -NR 13a COR 14a , -NR 13a CO 2 R 14A , -S(O)R 13A , -S(O) 2 R 13A , -SON(R 13A ) 2 , -NR 13A S(O) 2 R 14A ;-CSR 13A , -N(R 13A )R 14A , -C(O)N(R 13A )R 14A , -SO 2 N(R 13A )R 14A , and R 15A are independently selected from; R 13A and R 14A is hydrogen or R 15A are each independently selected from R 15A is a hydrocarbyl (e.g., C 1-6 alkyl, alkenyl, alkynyl, or haloalkyl), carbocyclyl, and -(CH 2 ) m1 -heterocyclyl, and each R 15A is halogen, cyano, amino, hydroxy, C 1-6 Alkyl or cycloalkyl, and C 1-6 optionally and independently substituted with one or more of alkoxy; m 1 is 0, 1, 2, 3, 4, 5, or 6; p 1 is 0, 1, 2, 3, or 4; R 4A The values ​​of may be the same or different; and q 1 is 0, 1, 2, 3, or 4; where R 5A The values ​​of may be the same or different; Y and Z, one or both of which may be absent, are hydrogen, R 16A , R 16A optionally substituted hydrocarbyl (e.g., C 1-6 alkyl, alkenyl, alkynyl, or haloalkyl), and R 16A optionally substituted with -(CH 2 ) r1 -heterocyclyl, wherein each R 16A is halogen, trifluoromethyl, cyano, thio, nitro, oxo, =NR 17A , -OR 17A , -SR 17A , -C(O)R 17A , -C(O)OR 17A , -OC(O)R 17A , -NR 17A COR 18A , -NR 17A CON(R 18A ) 2 , -NR 17A COR 18A , -NR 17A CO 2 R 18A , -S(O)R 17A , -S(O) 2 R 17A , -SON(R 17A ) 2 , -NR 17A S(O) 2 R 18A ;-CSR 17A , -N(R 17A )R 18A , -C(O)N(R 17A )R 18A , -SO 2 N(R 17A )R 18A , and R 19A are independently selected from; 1 is 0, 1, 2, 3, 4, 5, or 6; where: R 17A and R 18A is hydrogen or R 19A are each independently selected from R 19A is a hydrocarbyl (e.g., C 1-6 alkyl, alkenyl, alkynyl, or haloalkyl), carbocyclyl, and -(CH) s1 -heterocyclyl, and each R 19A is halogen, cyano, amino, hydroxy, C 1-6 Alkyl, and C 1-6 Optionally and independently substituted with one or more of alkoxy; and s 1 is 0, 1, 2, 3, 4, 5, or 6).

10. 10. The ADC of claim 9, wherein the NMT inhibitor is a compound of formula (IIIa): or a salt thereof 【Chemistry 6】 (In the formula: n 1 is 0 or 1; E 1 is C; W is a (1-4C)hydrocarbyl, aryl (e.g., phenyl), or heteroaryl group (e.g., pyridinyl); M is selected from C and N; R 3A , R 4A , and R 5A is hydrogen, R 12A , and R 12A independently selected from optionally substituted (1-3C)hydrocarbyl; R 12A is halogen, trifluoromethyl, cyano, thio, nitro, oxo, -OR 13A , -SR 13A , -C(O)R 13A , -C(O)OR 13A , -OC(O)R 13A , -NR 13A COR 14A , and R 15A are independently selected from; R 13A and R 14A are each independently selected from hydrogen or (1-4C)hydrocarbyl (e.g., methyl); Ring D * is an optionally substituted nitrogen-containing 6- or 7-membered heterocycle, wherein ring D * Each substitutable carbon or nitrogen in 7A optionally and independently substituted by; R 7A are independently selected from hydrogen, (1-4C)hydrocarbyl, halogen, trifluoromethyl, cyano, thio, nitro, or oxo; R 8A is hydrogen; p 1 is 0, 1, or 2, where R 4A The values ​​of may be the same or different; q 1 is 3, where R 5A The values ​​of may be the same or different; and t 1 is 0, 1, or 2, where R 7A The values ​​of may be the same or different).

11. The NMT inhibitor is (2,6-dichloro-4-(2-piperazin-1-yl-pyridin-4-yl)-N-(1,3,5-trimethyl-1H-pyraxol-4-yl)-benzenesulfonamide): 【Chemistry 7】 or a salt thereof.

12. The NMT inhibitor is 2,6-dichloro-N-(5-isobutyl-1,3-dimethyl-1H-pyrazol-4-yl(-4-(2-piperazin-1-yl-pyridin-4-yl)-benzenesulfonamide: 【Chemistry 8】 or a salt thereof.

13. The ADC or salt thereof according to any one of claims 1 to 4, wherein the NMT inhibitor is a compound of formula (V): 【Chemistry 9】 (In the formula: n 1 is 1 or 2; n 2 is 1 or 2; X 1 is CR x and N; If present, R x is hydrogen, halogen, and each substituent is halogen, -OH, -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 selected from the group consisting of alkyl; R 1 is hydrogen; each substituent is halogen, -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 alkyl; and each substituent is halogen, -CH 3 , -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 3-6 cycloalkyl; R 2 is hydrogen; each substituent is halogen, -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 alkyl; and each substituent is halogen, -CH 3 , -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 3-6 is selected from the group consisting of cycloalkyl; or R 1 and R 2 together with the atoms to which they are bonded, they are C 3-6 are linked to form a cycloalkyl group or a 3- to 6-membered non-aromatic heterocyclyl group containing one heteroatom selected from the group consisting of O and N, wherein the C 3-6 The cycloalkyl group or the 3- to 6-membered non-aromatic heterocyclyl group may each be substituted with halogen, -OH, -CH 3 , -OCH 3 , and -OCF 3 optionally substituted with one or two substituents independently selected from the group consisting of: R 3 is hydrogen; each substituent is halogen, -OH, -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 Alkyl; and each of the substituents is halogen, -OH, -CH 3 , -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 3-6- is selected from the group consisting of cycloalkyl; or R 1 and R 3 are linked together with the atoms to which they are attached such that they form a 3- to 6-membered non-aromatic heterocyclyl group containing one N heteroatom, wherein each substituent is selected from the group consisting of halogen, -CH 3 , -OH, -OCH 3 , and -OCF 3 optionally substituted with one or two substituents independently selected from the group consisting of: X 2 is CR 4 and N; If present, R 4 is hydrogen; halogen; each substituent is halogen, -OH, -OCH 3 , -OCF 3 , and -NR a R b -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 selected from the group consisting of alkyl; R 5a and R 5d is hydrogen; halogen; each substituent is halogen, -OH, -OCH 3 , and -OCF 3 Methyl optionally substituted with one, two, or three substituents independently selected from the group consisting of: 3 , and -OCF 3 methoxy optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of: R 5b and R 5c is hydrogen; halogen; each substituent is halogen, -OH, -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 Alkyl; each substituent is halogen, -OH, -OCH 3 , and -OCF 3 -OC optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 alkyl; and each substituent is halogen, -CH 3 , -OH, -OCH 3 , and -OCF 3 C optionally substituted with one, two, or three substituents independently selected from the group consisting of 3-6 independently selected from the group consisting of cycloalkyl; or R 5b and R 5c are linked together with the atoms to which they are attached such that they form a 6-membered aryl group or a 5- or 6-membered aromatic heterocyclyl group containing 1 or 2 heteroatoms selected from the group consisting of S, O, and N, wherein the 6-membered aryl group or the 5- or 6-membered aromatic heterocyclyl group is selected from the group consisting of halogen, —OH, —OCH 3 , and -OCF 3 optionally substituted with one or two substituents independently selected from the group consisting of: R 6 is selected from the group consisting of hydrogen and methyl; If present, each R 7 is a group in which each substituent is halogen, -OH, -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 is alkyl; R 8 is hydrogen; halogen; -OH; -CN; -C optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen, -OH, -CN, and methoxy optionally substituted with one, two, or three halogens. 1-4 Alkyl; each substituent is halogen, -CH 3 -C optionally substituted by one, two, or three substituents independently selected from the group consisting of -OH, -CN, and methoxy optionally substituted by one, two, or three halogens. 3-6 cycloalkyl; -C optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -OH, -CN, and methoxy optionally substituted with one, two, or three halogens; 1-4 alkenyl; and -OC optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of halogen, -OH, -CN, and methoxy optionally substituted with one, two, or three halogens. 1-4 selected from the group consisting of alkyl; R 9 is hydrogen and each substituent is halogen, -OH, -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 alkyl; or R 8 and R 9 together with the atoms to which they are attached, they form a six-membered aryl group, C 5-6 are linked to form a cycloalkyl group or a 5- or 6-membered aromatic heterocyclyl group containing 1 or 2 heteroatoms selected from N, O, and S, wherein the 6-membered aryl group, C 5-6 The cycloalkyl group or the 5- to 6-membered aromatic heterocyclyl group is optionally substituted with one, two, or three substituents, each of which is independently selected from the group consisting of halogen; -OH; -CN; and methoxy, each of which is optionally substituted with one, two, or three halogens. 1-4 alkyl; and -OC optionally substituted with one, two, or three substituents, each substituent independently selected from the group consisting of -halogen, -OH, and methoxy optionally substituted with one, two, or three halogens. 1-4 optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of alkyl; p is 0, 1, or 2; Z is a 5- to 13-membered non-aromatic heterocyclyl group containing 1, 2, or 3 heteroatoms selected from N, O, and S, where at least one of the heteroatoms is N, and where the 5- to 13-membered non-aromatic heterocyclyl group is selected from the group consisting of: each substituent being halogen; each substituent being halogen, —OH, and —OC optionally substituted with 1, 2, or 3 halogens. 1-3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 Alkyl; each substituent is halogen, -OH, and -OC optionally substituted with one, two, or three halogens. 1-3 -OC optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 Alkyl;NR c R d and -OC, each of which is optionally substituted with halogen, -OH, and one, two, or three halogens. 1-3 C optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 3-6 cycloalkyl; or when two substituents are at adjacent ring positions, they, together with the atoms to which they are attached, form a ring. 3-6 may be linked to form a cycloalkyl group or a 4- to 6-membered non-aromatic heterocyclyl group containing one heteroatom selected from the group consisting of O and N, wherein the C 3-6 The cycloalkyl group or the 4- to 6-membered non-aromatic heterocyclyl group is optionally substituted with one or two substituents, each of which is a halogen; each of which is a halogen, —OH, and —OC optionally substituted with one, two, or three halogens. 1-3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 alkyl; and -OC, each of which is optionally substituted with halogen, -OH, and one, two, or three halogens. 1-3 -OC optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 independently selected from the group consisting of alkyl; R c is hydrogen; R d is hydrogen; each substituent is halogen, -OCH 3 , and -OCF 3 -C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-6 alkyl; and each substituent is halogen, -CH 3 , -OH, -OCH 3 , and -OCF 3 C optionally substituted with one, two, or three substituents independently selected from the group consisting of 3-6 is selected from the group consisting of cycloalkyl; or Z is -NR 10 R 11 where: R 10 is hydrogen; and R 11 is a 5- to 10-membered non-aromatic heterocyclyl group containing 1, 2, or 3 heteroatoms selected from N, O, and S, where at least one of the heteroatoms is N, and where the 5- to 10-membered non-aromatic heterocyclyl group is selected from the group consisting of halogen; —OH; —OC, each of which is optionally substituted with halogen, —OH, and 1, 2, or 3 halogens. 1-3 -C optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of alkyl 1-6 alkyl; and —OC, each of the substituents being optionally substituted with halogen, —OH, and 1, 2, or 3 halogens. 1-3 -OC optionally substituted with one, two, or three substituents independently selected from the group consisting of alkyl 1-6 optionally substituted with 1, 2, 3, or 4 substituents independently selected from the group consisting of alkyl; and If present, each R a and R b is hydrogen and -C 1-4 alkyl).

14. 14. The ADC of claim 13, wherein the NMT inhibitor is a compound of formula (Vq): or a salt thereof. 【Chemistry 10】 (In the formula: R 3a is H or each substituent is halogen, -OCH 3 , and -OCF 3 C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 is alkyl; R 8a is halogen or each substituent is halogen, -OCH 3 , and -OCF 3 C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 is alkyl; ra is 0, 1, or 2; m a is 1 or 2; R 12a is hydrogen; and If present, each R 13a is a group in which each substituent is a halogen, -OCH 3 , and -OCF 3 C optionally substituted with one, two, or three substituents independently selected from the group consisting of 1-4 alkyl; and each substituent is halogen, -CH 3 , -OCH 3 , and -OCF 3 C optionally substituted with one, two, or three substituents independently selected from the group consisting of 3-6 independently selected from the group consisting of cycloalkyl; or ra is 2 and two R 13a When groups are located at adjacent ring positions, the two R 13a are each a halogen, -OH, and -OC optionally substituted with 1, 2, or 3 halogens, together with the atoms to which they are attached; 1-3 -C optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of alkyl 1-6 C optionally substituted with one or two substituents independently selected from the group consisting of alkyl 3-6 linked to form a cycloalkyl group).

15. The NMT inhibitor is (S)-1-(5-chloro-2-(2-methylpiperazin-1-yl)pyrimidin-4-yl)-N-(2-(imidazo[1,2-a]pyridin-3-yl)propan-2-yl)azetidine-3-carboxamide: 【Chemistry 11】 or a salt thereof.

16. The ADC or salt thereof according to any one of claims 1 to 4, wherein the NMT inhibitor is a compound of formula (VI): 【Chemistry 12】 (In the formula: R 1 is a group of formula OLA; L is -(CHR 12 ) m - and; Each R 12 are independently H or C 1-4 is alkyl; m is 1, 2, or 3; A is, 【Chemistry 13】 : and v is 0, 1, or 2; R 9a is H, C 1-4 Alkyl, or C 1-4 is haloalkyl; R 9b is H, C 1-4 Alkyl, or C 1-4 is haloalkyl; R 9c is C 1-4 Alkyl or C 1-4 is haloalkyl; R 9d is H, C 1-4 Alkyl, or C 1-4 is haloalkyl; R 10 is H, C 1-4 Alkyl, or C 1-4 is haloalkyl; R 11 H, halo, CN, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy or C 1-4 haloalkoxy; s is 0, 1, 2, or 3; Each R 2 are independently F, Cl, Br, C optionally substituted with up to three halogen groups 1-4 Alkyl, OCH 3 , or OCF 3 and; Y is CH or C 1-4 is alkyl; R 3 is H or C 1-4 is alkyl; R 4 is H or C 1-4 is alkyl; R 5 is H; R 6 is H or C 1-4 is alkyl; q is 0 or 1; R 7 is H or methyl; R 8 is H or methyl; or R 3 and R 6 and an intervening atom, the intervening atom and a bond, or the intervening atom and -(CHR a ) r -, or R 7 group and the R 6 The group and the intervening atom are separated by a -(CHR a ) r -, forming a 3- to 7-membered non-aromatic heterocycle consisting of; r is 1, 2, 3, 4, or 5; and R a is hydrogen or methyl).

17. 17. The ADC of claim 16, wherein the NMT inhibitor is a compound of formula (VIA): or a salt thereof 【Chemistry 14】 (In the formula: R 2a is H or F; R 2b is F; R 5a is H; R 6a is H or methyl; R 9ca is methyl, iso-propyl, or tert-butyl; R 9cb is H or methyl; R 10a is methyl; and R 11a is methyl; However, R 2a When is H, R 9cb is H).

18. The NMT inhibitor is 1-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}-2,2-dimethylpropan-1-ol: 【Chemistry 15】 or a salt thereof.

19. 2-{4-[2-(2,3-difluoro-6-{3-[(methylamino)methyl]imidazo[1,2-a]pyridin-6-yl}phenoxy)ethyl]-1,5-dimethyl-1H-pyrazol-3-yl}propan-2-ol: 【Chemistry 16】 or a salt thereof.

20. The ADC or salt thereof of any one of claims 1 to 19, wherein the linker is a cleavable linker.

21. 21. The ADC or salt thereof of claim 20, wherein the linker is cleaved by an enzyme.

22. The linker has the following formula (VII): -A a -W w -Y y -(Formula (VII)) (In the formula: A, if present, is a First Stretcher unit that forms a covalent bond with a chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of the antibody; a is 0 or 1; each W is independently an amino acid unit or a glucuronide unit that, when A and / or Y are absent, forms a covalent bond with a chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of the antibody and / or with a functional group of the NMT inhibitor, respectively; When W is an amino acid, w is 1 to 12; When W is a glucuronide unit, w is 1 or 2; Y, when present, is a second Stretcher unit that forms a covalent bond with a functional group of the NMT inhibitor; and y is 0 or 1) The ADC or salt thereof of any one of claims 1 to 21, having the following structure:

23. A is formula (A1): 【Chemistry 17】 (In the formula: n is 1 to 6; 【Chemistry 18】 represents the chain terminus (e.g., N-terminus) or point of attachment to a functional group of an amino acid side chain of the antibody; and 【Chemistry 19】 represents the point of attachment to W) 23. The ADC or salt thereof of claim 22, having the following structure:

24. A, 【Chemistry 20】 24. The ADC or salt thereof of claim 23, wherein:

25. A, 【Chemical 21】 23. The ADC or salt thereof of claim 22, wherein:

26. The ADC or salt thereof of any one of claims 22 to 25, wherein each W is an amino acid.

27. Each W has the formula (WI): 【Chemical 22】 wherein w is as defined above; and R 19 is H, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, CH 2 OH, CH(OH)Me, CH 2 CH 2 Sme, C.H. 2 C(O)Sme, CH 2 C(O)NH 2 , C.H. 2 C(O)OH, CH 2 CH 2 C(O)NH 2 , C.H. 2 CH 2 C(O)OH, (CH 2 ) 3 NHC(=NH)NH 2 , (CH 2 ) 3 NH 2 , (CH 2 ) 3 NHC(O)Me, (CH 2 ) 3 NHCHO, (CH 2 ) 4 NHC(=NH)NH 2 , (CH 2 ) 4 NH 2 , (CH 2 ) 4 NHC(O)Me, (CH 2 ) 4 NHCHO, (CH 2 ) 3 NHC(O)NH 2 , (CH 2 ) 4 NHC(O)NH 2 , C.H. 2 CH 2 CH(OH)CH 2 NH 2 , 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, phenyl, 【Chemical 23】 and; where: 【Chemistry 24】 represents the point of attachment to A (if present) or to the chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of the antibody; and 【Chemistry 25】 represents the point of attachment to Y (if present) or to a functional group of said NMT inhibitor 27. The ADC or salt thereof of claim 26,

28. At least one R 19 28. The ADC or salt thereof of claim 27, wherein is iso-propyl.

29. At least one R 19 (CH 2 ) 3 NHC(=NH)NH 2 28. The ADC or salt thereof of claim 27, wherein:

30. 30. The ADC or salt thereof of any one of claims 22 to 29, wherein w is 2.

31. (W)w is formula (WII): 【Chemical Formula 26】 (In the formula: R 19a is H, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, CH 2 OH, CH(OH)Me, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, phenyl, 【Chemical 27】 and R 19b is CH 2 CH 2 Sme, CH 2 C(O)Sme, CH 2 C(O)NH 2 , CH 2 C(O)OH, CH 2 CH 2 C(O)NH 2 , CH 2 CH 2 C(O)OH, (CH 2 ) 3 NHC(=NH)NH 2 , (CH 2 ) 3 NH 2 , (CH 2 ) 3 NHC(O)Me, (CH 2 ) 3 NHCHO, (CH 2 ) 4 NHC(=NH)NH 2 , (CH 2 ) 4 NH 2 , (CH 2 ) 4 NHC(O)Me, (CH 2 ) 4 NHCHO, (CH 2 ) 3 NHC(O)NH 2 , (CH 2 ) 4 NHC(O)NH 2 , or CH 2 CH 2 CH(OH)CH 2 NH<9000597>)(where) The ADC or salt thereof of any one of claims 22 to 30, having the following structure:

32. R 19a 32. The ADC or salt thereof of claim 31, wherein is iso-propyl.

33. R 19b (CH 2 ) 3 NHC(=NH)NH 2 33. The ADC or salt thereof of claim 31 or 32,

34. 23. The ADC or salt thereof of claim 22, wherein each W is a glucuronide unit.

35. (W)w is formula (WIII): 【Chemical Formula 28】 (In the formula: R is H, halo, CN, or NO 2 and; m is 0, 1, 2, or 3; Su is a sugar moiety; where: 【Chemical 29】 represents the point of attachment to A (if present) or to the chain terminus (e.g., N-terminus) or functional group of an amino acid side chain of the antibody; and 【Chemistry 30】 represents the point of attachment to Y (if present) or to a functional group of said NMT inhibitor 35. The ADC or salt thereof of claim 34, having the following structure:

36. 36. The ADC or salt thereof of claim 35, wherein m is 0.

37. Su has the following formula: 【Chemical 31】 37. The ADC or salt thereof of claim 35 or 36,

38. The ADC or salt thereof of any one of claims 34 to 37, wherein y is 0.

39. y is 1 and Y is a group of the formula (Y1): 【Chemical formula 32】 (In the formula: Each Q is independently halo, NO 2 , C.N., C. 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 haloalkoxy; m is 0 to 4; where: 【Chemical 33】 represents the point of attachment to W; and 【Chemical 34】 represents the point of attachment to the functional group of the NMT inhibitor. The ADC or salt thereof of any one of claims 22 to 33, having the following structure:

40. 40. The ADC or salt thereof of claim 39, wherein m is 0.

41. The linker has the formula (LI): 【Chemistry 35】 (In the formula, 【Chemical 36】 represents the point of attachment to a chain terminus (e.g., N-terminus) or functional group on an amino acid side chain of the antibody; and 【Chemical 37】 represents the point of attachment to the functional group of the NMT inhibitor.

23. The ADC or salt thereof of claim 22, having the following structure:

42. The linker has formula (LII): 【Chemical Formula 38】 (In the formula, 【Chemical 39】 represents the point of attachment to a chain terminus (e.g., N-terminus) or functional group on an amino acid side chain of the antibody; and 【Chemistry 40】 represents the point of attachment to the functional group of the NMT inhibitor.

23. The ADC of claim 22, comprising:

43. The linker has formula (LIII): 【Chemistry 41】 (In the formula, 【Chemistry 42】 represents the point of attachment to a chain terminus (e.g., N-terminus) or functional group on an amino acid side chain of the antibody; and 【Chemistry 43】 represents the point of attachment to the functional group of the NMT inhibitor.

22. The ADC of claim 20 or 21,

44. The linker has formula (LIV): 【Chemical 44】 (In the formula, 【Chemistry 45】 represents the point of attachment to a chain terminus (e.g., N-terminus) or functional group on an amino acid side chain of the antibody; and 【Chemistry 46】 represents the point of attachment to the functional group of the NMT inhibitor.

22. The ADC of claim 20 or 21,

45. The ADC or salt thereof of any one of claims 22 to 44, wherein the functional group on an amino acid side chain of the antibody is sulfhydryl.

46. 45. The ADC or salt thereof of any one of claims 22 to 44, wherein the functional group on the NMT inhibitor is amino or alcohol, e.g., amino.

47. The ADC or salt thereof according to any one of claims 1 to 46, wherein the antibody is a humanized antibody, a chimeric antibody, a human antibody, or an antibody fragment.

48. 48. The ADC or salt thereof of claim 47, wherein the antibody binds to HER2.

49. 49. The ADC or salt thereof of claim 48, wherein the antibody is trastuzumab, pertuzumab, margetuximab, ertumaxomab, MM-111, HER2Bi-aATCs, MCLA-128, ZW25, MDX-210, ado-trastuzumab, or fam-trastuzumab.

50. 50. The ADC or salt thereof of claim 49, wherein the antibody is trastuzumab.

51. 48. The ADC or salt thereof of claim 47, wherein the antibody binds to CD20.

52. 52. The ADC or salt thereof of claim 51, wherein the antibody is rituximab.

53. 48. The ADC or salt thereof of claim 47, wherein the antibody binds to Trop-2.

54. 54. The ADC or salt thereof of claim 53, wherein the antibody is sacituzumab.

55. 48. The ADC or salt thereof of claim 47, wherein the antibody binds to B7-H3.

56. 56. The ADC or salt thereof of claim 55, wherein the antibody is ifinatamab.

57. 57. The ADC or salt thereof of any one of claims 1 to 56, wherein the drug loading (p) of the antibody with NMT inhibitors is 1 to 10 NMT inhibitors per antibody, for example, 2 to 6, 4 to 6, 8 to 10, or 6 to 8 NMT inhibitors per antibody.

58. The ADC or a salt thereof has the following formula: 【Chemistry 47】 or a salt thereof, wherein Ab is the antibody of any one of claims 1 to 56, e.g., the antibody is trastuzumab, and p is the drug load of the NMT inhibitor of claim 57.

59. The ADC or a salt thereof has the following formula: 【Chemistry 48】 or a salt thereof, wherein Ab is the antibody of any one of claims 1 to 56, e.g., the antibody is trastuzumab, and p is the drug load of the NMT inhibitor of claim 57.

60. The ADC or a salt thereof has the following formula: 【Chemistry 49】 or a salt thereof, wherein Ab is sacituzumab and p is the drug load of the NMT inhibitor of claim 57.

61. The ADC or a salt thereof has the following formula: 【Chemistry 50】 or a salt thereof, wherein Ab is ifinatamab and p is the drug load of the NMT inhibitor of claim 57.

62. The ADC or a salt thereof has the following formula: 【Chemistry 51】 wherein Ab is the antibody of any one of claims 1 to 56, e.g., the antibody is trastuzumab, and p is the drug load of the NMT inhibitor of claim 57, or a salt thereof.

63. The ADC or a salt thereof has the following formula: 【Chemistry 52】 wherein Ab is the antibody of any one of claims 1 to 56, e.g., the antibody is trastuzumab, and p is the drug load of the NMT inhibitor of claim 57, or a salt thereof.

64. The ADC or a salt thereof has the following formula: 【Chemistry 53】 wherein Ab is the antibody of any one of claims 1 to 56, e.g., the antibody is trastuzumab, and p is the drug load of the NMT inhibitor of claim 57, or a salt thereof.

65. The ADC or a salt thereof has the following formula: 【Chemical 54】 wherein Ab is the antibody of any one of claims 1 to 56, e.g., the antibody is trastuzumab, and p is the drug load of the NMT inhibitor of claim 57, or a salt thereof.

66. 66. The ADC or salt thereof of any one of claims 58 to 65, wherein the ADC or salt thereof is attached to the antibody via a sulfhydryl group on the side chain of an amino acid on the antibody.

67. 67. A pharmaceutical composition comprising the ADC of any one of claims 3 to 66, or a pharmaceutically acceptable salt thereof.

68. 67. The ADC or a pharmaceutically acceptable salt thereof of any one of claims 3 to 66, for use as a pharmaceutical.

69. 69. The ADC or a pharmaceutically acceptable salt thereof of claim 68, for use in the prevention or treatment of a disease or disorder for which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect.

70. 67. Use of the ADC or a pharmaceutically acceptable salt thereof according to any one of claims 3 to 66 in the manufacture of a medicament for the prevention or treatment of a disease or disorder for which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect.

71. 67. A method for preventing or treating a disease or disorder in a subject for which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect, the method comprising administering a therapeutically effective amount of the ADC of any one of claims 3 to 66, or a pharmaceutically acceptable salt thereof.

72. 72. The ADC or a pharmaceutically acceptable salt thereof for use according to claim 69, the use according to claim 70, or the method according to claim 71, wherein the disease or disorder is a hyperproliferative disorder, and the hyperproliferative disorder is cancer.

73. 73. The ADC for use, or a pharmaceutically acceptable salt thereof, use, or method according to claim 72, wherein the cancer is colorectal cancer, gallbladder cancer, brain tumor, lymphoma (e.g., B-cell lymphoma or diffuse large B-cell lymphoma), leukemia (e.g., AML), or neuroblastoma.

74. 73. The ADC or a pharmaceutically acceptable salt thereof for use, use, or method according to claim 72, wherein the cancer is a hematological malignancy (e.g., a lymphoma, particularly a B-cell lymphoma (e.g., high-grade mantle zone lymphoma, follicular lymphoma, plasmablastic lymphoma, diffuse large B-cell lymphoma, and Burkitt's lymphoma), a myeloma (e.g., multiple myeloma), or a leukemia (e.g., chronic lymphocytic leukemia, AML, and B-acute lymphocytic leukemia)), or a solid tumor (e.g., brain cancer, lung cancer, breast cancer, prostate cancer, ovarian cancer, colorectal cancer, gallbladder cancer, kidney cancer, or liver cancer, or a blastoma (e.g., neuroblastoma, retinoblastoma, or glioblastoma)).

75. 73. The ADC for use or a pharmaceutically acceptable salt thereof, use, or method according to claim 72, wherein the cancer is selected from the group consisting of lung cancer, urothelial cancer, colorectal cancer, prostate cancer, ovarian cancer, pancreatic cancer, breast cancer, bladder cancer, gastric cancer, gastrointestinal stromal tumor, cervical cancer, esophageal cancer, squamous cell carcinoma, peritoneal cancer, liver cancer, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, vulvar cancer, thyroid cancer, and penile cancer, and expresses HER2 protein.

76. A drug conjugate, or a salt and / or solvate thereof, comprising an NMT inhibitor and a linker, wherein the linker comprises a group capable of forming a covalent bond with a chain terminus (e.g., N-terminus) or a functional group, e.g., a sulfhydryl group, on an amino acid side chain of an antibody.

77. formula: A a -W w -Y y -NMT (Formula (X)) (In the formula: A, if present, is a First Stretcher unit that contains a group that can form a covalent bond with a chain terminus (e.g., N-terminus) or a functional group, e.g., a sulfhydryl group, on an amino acid side chain of an antibody; The NMT is an NMT inhibitor according to any one of claims 1 to 19; and a, W, w, Y, and y are as defined in any one of claims 22 to 59. or a salt and / or solvate thereof.

78. A, 【Chemistry 55】 (where, 【Chemical 56】 represents the point of attachment to W) 78. The drug conjugate of claim 77, its salt and / or solvate,

79. The drug conjugate has the formula (DC-2): 【Chemical 57】 wherein NMT is an NMT inhibitor according to any one of claims 1 to 19, in particular an NMT inhibitor according to any one of claims 16 to 19. or a salt and / or solvate thereof.

80. (1S,2R,3S,4R,5R)-5-(4-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-[3-(3-{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy]ethoxy}propanamido)propanamido]phenoxy)-3,4-dihydroxy-2-methylcyclohexane-1-carboxylic acid: 【Chemistry 58】 80. The drug conjugate according to any one of claims 76 to 79, or a salt and / or solvate thereof.

81. 【Catalog 59】 80. The drug conjugate according to any one of claims 76 to 79, or a salt and / or solvate thereof.

82. The drug conjugate has the formula (DC-1): 【Chemistry 60】 wherein NMT is an NMT inhibitor according to any one of claims 1 to 19, in particular an NMT inhibitor according to any one of claims 16 to 19. or a salt and / or solvate thereof.

83. {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido]-3-methylbutanamido]pentanamido]phenyl}methyl N-{[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-2,2-dimethylpropyl)-1,5-dimethyl-1H-pyrazol-4-yl]ethoxy}phenyl)imidazo[1,2-a]pyridin-3-yl]methyl}-N-methylcarbamate: 【Hua 61】 83. The drug conjugate, salt thereof and / or solvate thereof according to claim 76, 77, or 82, wherein 【Request 84】 【Chemical 62】 80. The drug conjugate according to any one of claims 76 to 79, or a salt and / or solvate thereof.

85. The drug conjugate has the formula (DC-5): 【Chemistry 63】 wherein NMT is an NMT inhibitor according to any one of claims 1 to 19, in particular an NMT inhibitor according to any one of claims 16 to 19. or a salt and / or solvate thereof. 【Request 86】 【Chemical 64】 or a salt and / or solvate thereof.

87. The drug conjugate has the formula (DC-6): 【Chemistry 65】 wherein NMT is an NMT inhibitor according to any one of claims 1 to 19, in particular an NMT inhibitor according to any one of claims 16 to 19. or a salt and / or solvate thereof.

88. 88. A salt of the drug conjugate of any one of claims 76 to 87.

89. A solvate of the drug conjugate of any one of claims 76 to 87.

90. 88. A solvate of the salt of the drug conjugate of any one of claims 76 to 87.

91. The drug conjugate of any one of claims 76 to 87.

92. Compounds of formula (ADC-I): 【Hua 66】 and Compound of formula (ADC-II): 【Chemical 67】 wherein NMT is an NMT inhibitor according to any one of claims 1 to 19, in particular an NMT inhibitor according to any one of claims 16 to 19.

10. A compound selected from the group consisting of: or a salt and / or solvate of any one of these.

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