Antibody drug conjugates comprising NMT inhibitors and uses thereof

Abstract

The present invention relates to ADCs comprising an NMT inhibitor conjugated to an antibody via a linker, 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 in particular to the use of such ADCs as medicaments, in particular for the treatment or prevention of hyperproliferative disorders such as cancer. Background Art

[0002] Antibody therapy has been demonstrated to be useful for targeted treatment of patients with cancer, immune and angiogenic disorders (Carter, P. (2006) Nature Reviews Immunology 6:343-357). Antibody drug conjugates (ADCs), i.e., immunoconjugates, are used to locally deliver cytotoxic or cytostatic agents, i.e., drugs that kill or inhibit tumor cells in cancer treatment, targeting the drug moiety to the tumor and accumulating within the cells therein, whereas systemic administration of these unconjugated drug formulations may produce 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.Patents15(9):1087-1103; Payne, G. (2003) Cancer Cell 3:207-212; Trail et al. (2003) CancerImmunol.Immunother.52:328-337; Syrigos and Epenetos (1999) Anticancer Research 19:605-614).

[0003] Thus, the maximum efficacy and minimum toxicity are sought. Efforts to design and improve ADCs have focused on the selectivity of monoclonal antibodies (mAbs) as well as drug action mechanisms, drug linkage, drug / antibody ratio (drug 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). The drug moiety can exert cytotoxic and cytostatic effects through mechanisms such as tubulin binding, DNA binding, proteasome and / or topoisomerase inhibition. Some cytotoxic drugs are often inactive or less active when coupled to large antibodies or protein receptor ligands.

[0004] N-myristoyltransferase (NMT) is a monomeric enzyme that is ubiquitous in eukaryotes. NMT catalyzes the irreversible co-translational transfer of myristic acid (a saturated 14-carbon fatty acid) from myristoyl-CoA (myr-CoA) to protein substrates containing an N-terminal glycine, and forms an amide bond (Farazi, TA, G. Waksman and J. I. Gordon, J. Biol. Chem., 2001. 276 (43): p. 39501-39504).

[0005] There are two types of human NMT, human NMT1 (HsNMT1) and human NMT2 (HsNMT2). Inhibition of human NMT has been suggested as a target for treating or preventing various diseases or disorders, such as hyperproliferative disorders (e.g., cancer, such as human colorectal cancer, gallbladder cancer, brain tumors, and lymphomas, such as B-cell lymphomas) (Resh MD. 1993. Biochern. Biophys. Acta 1115, 307-22; Bertiaume LG, Beuachamp E, WO2017011907), and 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 LP, Killington, RA, Rowlands DJ,Tuthill,TJ,2008Journal ofVirology 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,Hopkins-Navratilova I,Wilkinson AJ,Johnston SL,Leatherbarrow RJ,Tuthill TJ,Solari R,Tate EW 2018Nature Chemistry 10(6)599-606),Corbic RamljakI,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. Since NMTs play key 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 multi-protein pathways.Although human NMT inhibitors are expected to inhibit both HsNMT1 and HsNMT2, their therapeutic and / or prophylactic activity is believed to arise primarily from inhibition of HsNMT1. Such properties are believed to be advantageous in reducing the risk of resistance development, for example in the treatment or prevention of microbial infections and hyperproliferative disorders.

[0006] There are two known binding pockets in NMT. ​​One is the myr-CoA binding pocket and the other is the peptide binding pocket. Most of the NMT inhibitors reported so far target the peptide binding pocket.

[0007] Compounds that can be used as NMT inhibitors have been previously disclosed, for example, see WO00 / 37464 (Roche), WO2010 / 026365 (University of Dundee), WO2013 / 083991 (ImperialInnovations 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, etc.). Specific uses of NMT inhibitors have been disclosed, for example, see WO2022 / 090746 (Imperial College Innovations Limited, etc.) and WO2022 / 082306 (Pacylex Pharmaceuticals Inc.).

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

[0009] Surprisingly, the inventors have now found that ADCs containing NMT inhibitors exhibit potent cytotoxic activity with minimal adverse side effects (e.g., weight loss). These properties are expected to make the ADCs of the invention particularly suitable for use as drugs for the treatment or prevention of hyperproliferative diseases such as cancer. Summary of the invention

[0010] The present invention provides an antibody-drug conjugate (ADC), which comprises an NMT inhibitor coupled to an antibody via a linker, or a salt thereof (referred to herein as "the ADC of the present invention").

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

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

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

[0014] The present invention also provides an ADC of the present invention or a pharmaceutically acceptable salt thereof for use in preventing or treating a disease or condition (eg, cancer) in which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect.

[0015] The present invention also provides the use of the ADC of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing or treating a disease or condition (eg, cancer) in which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect.

[0016] The present invention also provides a method for preventing or treating a disease or condition (e.g., cancer) in a subject in which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect, the method comprising administering a therapeutically effective amount of an ADC of the present invention or a pharmaceutically acceptable salt thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 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.

[0018] Figure 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.

[0019] Figure 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.

[0020] Figure 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.

[0021] 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.

[0022] Figure 6 : Shows the effect of treatment with 5 mg / Kg trastuzumab, 5 mg / Kg ADC Example 1, 5 mg / Kg trastuzumab and 5 mg / Kg isotype control antibody on tumor volume in a mouse gastric cancer xenograft model.

[0023] Fig. 7A : Show Biological Example 3 (and Figure 5 , 2.5 mg / kg (mpk)).

[0024] Figure 7B : Show Biological Example 3 (and Figure 6 , 5 mg / kg (mpk)).

[0025] Figure 8-Figure 11 : Shows the conjugate of trastuzumab and NMT inhibitor 1 (ADC Example 1; Figure 8 ), trastuzumab ( Fig. 9 ), trastuzumab ( Fig.10 ) and isotype controls (conjugate of isotype control IgG and NMT inhibitor 1; Fig.11 ) In vitro cytotoxicity evaluation results against HER2-positive breast cancer cell line BT474. Puromycin was used as a positive cytotoxicity control.

[0026] Figure 12-Figure 15 : Shows the conjugate of trastuzumab and NMT inhibitor 1 (ADC Example 1; Fig.12 ), trastuzumab ( Fig.13 ), trastuzumab ( Fig.14 ) and isotype controls (conjugate of isotype control IgG and NMT inhibitor 1; Fig.15 ) In vitro evaluation of cytotoxicity against the HER2-negative breast cancer cell line MCF7. Puromycin was used as a positive cytotoxicity control.

[0027] Fig.16: Shows the in vitro "bystander effect" results of different concentrations (12.5nM, 3.13nM, 0.78nM, 0.2nM and 0.05nM) of trastuzumab conjugate with NMT inhibitor 1 (ADC Example 1), trastuzumab, Reference Example 1 (a conjugate of trastuzumab with monomethyl auristatin E (MMAE)), isotype control and trastuzumab on the HER2-negative breast cancer cell line MCF7.

[0028] Fig.17 : Shows the in vitro "bystander effect" results of different concentrations (12.5nM, 3.13nM, 0.78nM, 0.2nM and 0.05nM) of trastuzumab conjugate with NMT inhibitor 1 (ADC Example 1), trastuzumab, Reference Example 1 (a conjugate of trastuzumab with monomethyl auristatin E (MMAE)), isotype control and trastuzumab on the HER2-positive breast cancer cell line BT474.

[0029] Figure 18-Figure 27 : ADC Examples 1-5 are shown to be effective against a panel of cell lines including BT474 ( Fig.18 )、JIMT 1( Fig.19 )、NCI N87( Fig. 20 )、NCI H292( Fig.21 )、IM95-m( Fig. 22 )、ZR-75-30( Fig.23 )、NCI H2170( Fig.24 )、LNCaP( Fig.25 )、C42( Fig.26 ) and VCaP( Fig. 27 )) in vitro cytotoxicity assessment results, wherein various comparators (e.g., sacituzumab, trastuzumab, ifinatamab, NMT inhibitor 1, and isotype control) are provided in each figure.

[0030] Figure 28-Figure 43 : The results of the effects of the test article isotype control, ADC Example 1, staurosporine, trastuzumab, and detrastuzumab as single agents on gastric cancer organoid cell viability using the CellTiter-Glo (CTG) luminescent cell viability assay are shown as follows: GA0429B ( Fig.28 )、GA6877B( Fig.29 )、GA6894B( Fig.30 )、GA2434B( Fig.31 )、GA3102B( Fig.32 )、GA0119B( Fig.33 )、GA0091B( Fig.34 )、GA6815B( Fig.35 )、GA0098B( Fig.36 )、GA6833B( Fig.37 )、GA0087B( Fig.38 )、GA2109B( Fig.39 )、GA3055B( Fig.40 )、GA6866B( Fig.41 )、GA0060B( Fig.42 ) and GA6891B( Fig.43 ).

[0031] Fig.44a - b: Shows the mean (+ / - SEM) levels of hematological markers in the 20 mg / Kg group described in Biological Example 9, normalized to baseline (pre-dose) values.

[0032] Fig.45 : Shows the weight loss of mice in Biological Example 10.

[0033] Fig.46 : Shows the average values ​​(from left to right) of AST (aspartate aminotransferase), ALT (alanine aminotransferase), ALP (alkaline phosphatase), LDH (lactate dehydrogenase), CK (creatine kinase) and GGT (γ-glutamyl transferase) in the blood of animals for each dose at the end of Biological Example 10.

[0034] Fig.47 : Shows the mean values ​​of LYM, MON, NEU, RBC, HGB and PLT cells in the blood of animals for each dose at the end of Biological Example 10 (from left to right).

[0035] Fig.48 : Shows the effect of treatment with ifenatumomab (5 mg / kg and 10 mg / kg), ifenatumomab-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.

[0036] Fig.49 : Shows the effect of treatment on body weight with ifenatumab (5 mg / kg and 10 mg / kg), ifenatumab-DXd (5 mg / kg and 10 mg / kg), ADC Example 4 (5 mg / kg and 10 mg / kg), and vehicle control in a mouse LNCaP prostate cancer xenograft model.

[0037] Fig.50: Shows the effect of treatment with ifenatumomab (5 mg / kg), ifenatumomab-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.

[0038] Fig.51 : Shows the effect of treatment on body weight with ifenatumab (5 mg / kg), ifenatumab-DXd (5 mg / kg), ADC Example 4 (2.5 mg / kg, 5 mg / kg and 10 mg / kg) and vehicle control in a mouse VCaP prostate cancer xenograft model.

[0039] Fig.52 : Shows the effect of treatment on tumor volume with sacituzumab (5 mg / kg), sacituzumab govitecan (5 mg / kg) (plus ADC Example 3, 5 mg / kg added on study days 27 and 34), and ADC Example 3 (5 mg / kg) and vehicle control in the mouse JIMT-1 breast cancer xenograft model.

[0040] Fig.53 : Shows the effect of treatment on tumor volume with sacizumab (2.5 mg / kg), sacizumab-govitecan (2.5 mg / kg, plus ADC Example 3, 5 mg / kg added on study day 27), and ADC Example 3 (2.5 mg / kg) and vehicle control in the mouse JIMT-1 breast cancer xenograft model.

[0041] Fig.54 : Shows the effect of treatment on body weight in the mouse JIMT-1 breast cancer xenograft model with sacizumab (5 mg / kg), sacizumab-govitecan (5 mg / kg) (plus ADC Example 3, 5 mg / kg added on study days 27 and 34), and ADC Example 3 (5 mg / kg and 10 mg / kg) and vehicle control.

[0042] Fig.55 : Shows the effect of treatment on body weight with sacizumab (2.5 mg / kg), sacizumab-govitecan (2.5 mg / kg, plus ADC Example 3, 5 mg / kg added on study day 27), and ADC Example 3 (2.5 mg / kg) and vehicle control in the mouse JIMT-1 breast cancer xenograft model.

[0043] Fig.56: Shows the results of in vitro evaluation of the cytotoxicity of the conjugate of trastuzumab and NMT inhibitor 1 (ADC Example 8) at different concentrations (50nM, 3.13nM, 0.2nM and 0nM) against the HER2-positive breast cancer cell line BT474. Puromycin was used as a positive cytotoxicity control.

[0044] Fig.57 : Shows the results of in vitro evaluation of the cytotoxicity of the conjugate of trastuzumab and NMT inhibitor 1 (ADC Example 8) at different concentrations (50nM, 3.13nM, 0.2nM and 0nM) against the HER2-negative breast cancer cell line MCF7. Puromycin was used as a positive cytotoxicity control.

[0045] Sequence Listing

[0046] SEQ ID NO: 1 - Amino acid sequence of trastuzumab light chain

[0047] SEQ ID NO:2—Amino acid sequence of trastuzumab heavy chain

[0048] SEQ ID NO: 3—Amino acid sequence of rituximab light chain

[0049] SEQ ID NO: 4—Amino acid sequence of rituximab heavy chain

[0050] SEQ ID NO:5—Amino acid sequence of the ifenatumomab light chain

[0051] SEQ ID NO:6—Amino acid sequence of ifenatumomab heavy chain

[0052] SEQ ID NO:7—Amino acid sequence of the light chain of tadalafil

[0053] SEQ ID NO:8—Amino acid sequence of the heavy chain of tadalafil DETAILED DESCRIPTION

[0054] The term "hydrocarbyl" as used herein is understood to mean a saturated, unsaturated or partially unsaturated hydrocarbon group of any compound, straight or branched. Suitable examples of "hydrocarbyl" groups may include, for example, "alkyl", "alkenyl", "alkynyl" and / or "haloalkyl" groups, each of which is defined below.

[0055] The term "alkyl" as used herein is understood to mean straight and branched saturated hydrocarbon groups. Examples of "alkyl" groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, pentyl and hexyl. Among unbranched alkyl groups, methyl, ethyl, n-propyl, isopropyl, n-butyl are preferred. Among branched alkyl groups, tert-butyl, isobutyl, 1-ethylpropyl and 1-ethylbutyl may be mentioned.

[0056] The term "C m-n " or "(m-nC) group" when used alone or as a prefix refers to any group having m to n carbon atoms.

[0057] The term "alkenyl" as used herein means straight and branched 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.

[0058] The term "alkynyl" as used herein means straight and branched 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.

[0059] As used herein, the term "carbocyclyl" (or "carbocycle") means any 3 to 13 membered carbocyclic ring system, which may be saturated, partially unsaturated or aromatic. The carbocyclic ring system may be monocyclic or may contain more than one ring (e.g., the ring system may be bicyclic). Examples of monocyclic saturated carbocyclic rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl. Examples of bicyclic saturated carbocyclic rings include bicyclooctane, bicyclononane, bicyclodecane (decalin), and bicyclooctane. Another example of a saturated carbocyclic ring is adamantane. Examples of monocyclic unsaturated carbocyclic rings include cyclobutene, cyclopentene, cyclopentadiene, cyclohexene. Examples of aromatic carbocyclic rings include phenyl and naphthyl. Further examples of aromatic carbocyclic rings include tetrahydronaphthyl (tetralin) and indan.

[0060] The term "cycloalkyl" as used herein means a saturated group in a ring system. The cycloalkyl group may be monocyclic or bicyclic. For example, the bicyclic group may be 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.

[0061] The term "halogen" or "halo" as used herein means fluorine, chlorine, bromine or iodine. Fluorine, chlorine and bromine are particularly preferred.

[0062] As used herein, the term "haloalkyl" means an alkyl group having a halogen substituent, and the terms "alkyl" and "halogen" are understood to have the above meanings. Similarly, the term "dihaloalkyl" means an alkyl group having two halogen substituents, and the term "trihaloalkyl" means an alkyl group having three halogen substituents. Examples of haloalkyl groups include fluoromethyl, chloromethyl, bromomethyl, fluoromethyl, fluoropropyl and fluorobutyl; examples of dihaloalkyl groups include difluoromethyl and difluoroethyl; examples of trihaloalkyl groups include trifluoromethyl and trifluoroethyl.

[0063] The term "heterocyclyl" (or heterocycle) as used herein means an aromatic or non-aromatic carbon atom cyclic group in which one to four carbon atoms are replaced by one or more heteroatoms independently selected from nitrogen, oxygen or sulfur. A heterocyclyl (or heterocyclic) group can be, for example, a monocyclic or bicyclic ring. In a bicyclic heterocyclyl (or heterocyclic) group, each ring can have one or more heteroatoms, or heteroatoms can be present in only one of the rings. The heteroatom can be S, O or N, preferably O or N.

[0064] Examples of monocyclic non-aromatic heterocyclic groups (or heterocycles) include aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, and azepanyl.

[0065] Examples of monocyclic aromatic heterocyclic (or heterocyclic) groups include furanyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, triazolyl, triazinyl, tetrazolyl, pyridazinyl, isothiazolyl, isoxazolyl, pyrazinyl, pyrazolyl and pyrimidinyl.

[0066] Examples of bicyclic aromatic heterocyclyls (or heterocycles) include quinoxalinyl, quinazolinyl, pyridopyrazinyl, benzoxazolyl, benzothiophenyl, benzimidazolyl, naphthyridinyl, quinolinyl, benzofuranyl, indolyl, benzothiazolyl, oxazolyl [4,5-b] pyridinyl, pyridopyrimidinyl, isoquinolinyl, and benzodioxazole. Further examples of bicyclic aromatic heterocyclyl groups include those in which one ring is aromatic and the other ring is non-aromatic, such as dihydrobenzofuranyl, indanyl, indolinyl, isoindolinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, and benzazepanyl.

[0067] The term "optionally substituted" refers to substituted as well as unsubstituted groups, structures or molecules. The term "wherein R 1 One or any CH, CH2, CH3 group or heteroatom (i.e. NH) in the group is optionally substituted" suitably means R 1 (Any) one of the hydrogen radicals of a group is replaced by the relevant specified group.

[0068] When optional substituents are selected from "one or more" groups, it is understood that this definition includes all substituents being selected from one of the specified groups, or substituents being selected from two or more of the specified groups.

[0069] NMT inhibitors

[0070] As described above, the ADC of the present invention comprises an NMT inhibitor. In one embodiment, the NMT inhibitor is a compound of formula (I):

[0071]

[0072] or a salt thereof,

[0073] in:

[0074] Y is selected from -CH-, -C(R 2 )- and -N-;

[0075] R 1 is a group of formula –X–L–A;

[0076] X stands for -O-;

[0077] L stands for -(CH2) m -;

[0078] m is 1, 2 or 3;

[0079] A is a 6-10 membered aromatic carbocyclic ring or a 5-10 membered aromatic heterocyclic ring, the aromatic carbocyclic ring or heterocyclic ring is optionally substituted by 1, 2 or 3 substituents, each substituent being independently selected from the group consisting of: -F, -Cl, -Br, -OCH3, -OCF3, -CN,

[0080] Optionally with up to 3 halogen, hydroxyl or -OC 1-4 Alkyl-substituted-C 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)2, -CH2C(O)N(R 9 )2、-CH2C(O)N(R 13 )C 1-4 Alkyl OC 1-4 Alkyl, -CH2C(O)N(C 1-4 Alkyl OC 1-4Alkyl)2, -S(O)2NHC 1-4 Alkyl, -S(O)2N(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, CH2N(R 13 )2、CH2N(R 13 )C(O)C 1-4 Alkyl, CH2N(R 13 )S(O)2C 1-4 Alkyl, -CH2S(O)2C 1-4 Alkyl and CO2H;

[0081] s is 0, 1, 2, or 3;

[0082] Each R 2 independently selected from the group consisting of: -F, -Cl, -Br, -OCH3, -OCF3, -CN, -C optionally substituted with up to 3 halogens 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)2N(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 alkyl;

[0083] q is 0 or 1;

[0084] R 3 is hydrogen or methyl; R 4 is hydrogen or methyl;

[0085] R 5 is hydrogen; R 6 is hydrogen or C optionally substituted by up to 3 -F, -Cl, -Br, -OH, -OCH3, -OCF3 or -CN groups 1-6 alkyl;

[0086] When present, R 10 is hydrogen or methyl;

[0087] When present, R 11 is hydrogen or methyl;

[0088] or R3 Group and R 5 The groups and the atoms therebetween form bonds between the atoms therebetween, or between the atoms and -(CHR a ) r - a 3- to 7-membered non-aromatic heterocyclic ring; or R 10 Group and R 5 The groups and the atoms between them form the atoms between them and -(CHR a ) r -a 3- to 7-membered non-aromatic heterocyclic ring;

[0089] r is 1, 2, 3, 4 or 5; R a is hydrogen or methyl;

[0090] Each R 7 are independently selected from hydrogen, halogen, C 1-4 Alkoxy and optionally substituted by 1, 2 or 3 halogen 1-4 alkyl; and

[0091] R 8 Selected from hydrogen and C 1-4 alkyl;

[0092] Each R 9 independently selected from hydrogen and C 1-4 Alkyl groups, or two R 9 The groups and the N to which they are bound form a 4- to 7-membered non-aromatic heterocyclic ring, the heterocyclic ring optionally containing 1 or 2 further heteroatoms selected from N, O and S;

[0093] Each R 13 independently selected from hydrogen and C 1-4 alkyl; and wherein

[0094] i) E, J and G are each C(R 7 ), K is carbon, Q is N(R 8 ), and M is nitrogen;

[0095] ii) E, J and G are each C(R 7 ), and K, Q and M are each nitrogen; or

[0096] iii) E, J, G and M are each C(R 7 ), and K and Q are each nitrogen.

[0097] Suitably, the compound of formula (I) or a salt thereof is 5 )(R 6 ) group is attached to the linker.

[0098] In one embodiment, the NMT inhibitor is a compound of formula (IA^^):

[0099]

[0100] or a salt thereof,

[0101] in:

[0102] R 1 is a group of formula -XLA;

[0103] A is 4-pyrazolyl, which is optionally substituted by up to 3 groups selected from methyl and -C(O)N(CH3) 2的 Substituent substitution;

[0104] X is -O-;

[0105] L is –(CH2) m -;

[0106] m is 2;

[0107] R 2′ Selected from the group consisting of fluorine or chlorine (preferably fluorine);

[0108] R 2″ selected from the group consisting of hydrogen, fluorine or chlorine;

[0109] q is 0;

[0110] R 3 is hydrogen or methyl;

[0111] R 4 is hydrogen or methyl;

[0112] R 5 is hydrogen;

[0113] R 6 is hydrogen or methyl; or

[0114] R 3 Group and R 6 The groups and the atoms therebetween form a 3- to 7-membered non-aromatic heterocyclic ring consisting of the atoms and bonds therebetween;

[0115] E, J, G, K, Q and M are:

[0116] i) E, J and G are each CH, K is carbon, Q is N (R 8 ), M is nitrogen; and R 8 is hydrogen or methyl; or

[0117] ii) E, J, G and M are each CH, and K and Q are each nitrogen;

[0118] Provided that A is substituted with not more than one -C(O)N(CH3)2 group.

[0119] Suitably, the compound of formula (IA^^) or a salt thereof is 5 )(R 6 ) group is attached to the linker.

[0120] Compounds of formula (I) and formula (IA) are disclosed in WO2017 / 001812, the entire contents of which are incorporated herein by reference to describe the synthesis and activity of NMT inhibitors.

[0121] In one embodiment, the NMT inhibitor is a compound of formula (II):

[0122]

[0123] or a salt thereof,

[0124] in:

[0125] R 1 is H or -CH3; and

[0126] R 2 It is H or F.

[0127] Suitably, the compound of formula (II) or a salt thereof is linked to the linker via an NH2 group.

[0128] 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:

[0129]

[0130] or a salt thereof.

[0131] Suitably, 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 linked to the linker via an NH2 group.

[0132] Compounds 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 herein by reference to describe the synthesis and activity of NMT inhibitors.

[0133] In one embodiment, the NMT inhibitor is a compound of formula (III) or (IV):

[0134]

[0135] or a salt thereof,

[0136] in:

[0137] n1 is 0, 1, 2, 3, 4, 5 or 6;

[0138] Ring A* is an optionally substituted nitrogen-containing aryl group, wherein each substitutable carbon or nitrogen in Ring A* is optionally and independently replaced by one or more R 5A and wherein if ring A* contains an -NH- moiety, the nitrogen may be optionally replaced by C 1-6 alkyl (eg, methyl) substituted; and wherein R 4A and ring A* may form a cyclic group together with the atoms to which they are attached,

[0139] Ring B* is an optionally substituted aryl or heteroaryl group, wherein each substitutable carbon or heteroatom in Ring B* is optionally and independently replaced by one or more R 3A replace;

[0140] One of W and X may not be present, and W and X are independently selected from R 11A , optionally by R 11A Substituted hydrocarbon groups (e.g., C 1-8 alkyl, alkenyl, alkynyl or haloalkyl) and optionally R 12A Substituted -(CH2) k1 -heterocyclyl; k1 is 0, 1, 2, 3, 4, 5 or 6;

[0141] R 1A is hydrogen;

[0142] R 2A , R 3A , R 4A and R 5A are independently selected from hydrogen, R 12A , optionally by R 12A Substituted hydrocarbon groups (e.g., C 1-6 alkyl, alkenyl, alkynyl or haloalkyl) and optionally substituted by one or more R 12A Substituted -(CH2) L1 -heterocyclic group; wherein R 2A Together with W or X, they may form a 12A substituted heterocycle; and wherein R 3A and R 5A One or more of the atoms to which they are attached may form an optionally R 12A Substituted carbocyclic ring, such as heterocyclic ring; L1 is 0, 1, 2, 3, 4, 5 or 6;

[0143] in:

[0144] Each R 11A and R 12A are independently selected from 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 ;

[0145] R 13A and R 14A are each independently selected from hydrogen or R 15A ;

[0146] R 15A Selected from hydrocarbon groups (such as C 1-6 alkyl, alkenyl, alkynyl or haloalkyl), carbocyclic group and -(CH2) m1 -heterocyclic group, and each R 15A Optionally and independently halogen, cyano, amino, hydroxyl, C 1-6 Alkyl or cycloalkyl and C 1-6 one or more substitutions in the alkoxy group;

[0147] m1 is 0, 1, 2, 3, 4, 5 or 6;

[0148] p1 is 0, 1, 2, 3 or 4; R 4A The values ​​of can be the same or different; and

[0149] q1 is 0, 1, 2, 3 or 4; where R5A The values ​​of can be the same or different;

[0150] One or both of Y and Z may be absent and are independently selected from hydrogen, R 16A , optionally by R 16A Substituted hydrocarbon groups (such as C 1-6 alkyl, alkenyl, alkynyl or haloalkyl) and optionally R 16A Substituted -(CH2) r1 -heterocyclic group, wherein each R 16A are independently selected from 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;

[0151] in:

[0152] R 17A and R 18A are each independently selected from hydrogen or R 19A ;

[0153] R 19A Selected from hydrocarbon groups (such as C 1-6 alkyl, alkenyl, alkynyl or haloalkyl), carbocyclic group and -(CH2) s1 -heterocyclic group, and each R 19A Optionally and independently halogen, cyano, amino, hydroxyl, C 1-6 Alkyl and C1-6 one or more substitutions in the alkoxy group; and

[0154] s1 is 0, 1, 2, 3, 4, 5, or 6.

[0155] Suitably, the compound of formula (III) or a salt thereof is 1A )(R 2A ) group is connected to the linker. Alternatively, when Z is -N(R 17A )R 18A When the compound of formula (IV) or a salt thereof is suitably linked to the linker via Z, or when Z is absent and Y is -N(R 17A )R 18A When Y is used, it is suitably connected to the linker via Y.

[0156] In one embodiment, the NMT inhibitor is a compound of formula (IIIa):

[0157]

[0158] or a salt thereof,

[0159] in:

[0160] n1 is 0 or 1;

[0161] E 1 is C;

[0162] W is a (1-4C)alkyl, aryl (e.g. phenyl) or heteroaryl group (e.g. pyridyl);

[0163] M is selected from C and N;

[0164] R 3A , R 4A and R 5A are independently selected from hydrogen, R 12A and optionally R 12A substituted (1-3C)alkyl;

[0165] R 12A are independently selected from 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 ;

[0166] R 13A and R 14Aare each independently selected from hydrogen or (1-4C)hydrocarbyl (eg methyl);

[0167] Ring D* is an optionally substituted nitrogen-containing 6- or 7-membered heterocyclic ring, wherein each substitutable carbon or nitrogen in Ring D* is optionally and independently replaced by one or more R 7A replace;

[0168] R 7A independently selected from hydrogen, (1-4C)alkyl, halogen, trifluoromethyl, cyano, thio, nitro or oxo;

[0169] R 8A is hydrogen;

[0170] p1 is 0, 1 or 2, where R 4A The values ​​of can be the same or different;

[0171] q1 is 3, where R 5A The values ​​of can be the same or different; and

[0172] t1 is 0, 1 or 2, where R 7A The values ​​can be the same or different.

[0173] Suitably, the compound of formula (IIIa) or a salt thereof is prepared by NR 8A The group is attached to the linker.

[0174] In one embodiment, the NMT inhibitor is (2,6-dichloro-4-(2-piperazin-1-yl-pyridin-4-yl)-N-(1,3,5-trimethyl-1H-pyrazol-4-yl)-benzenesulfonamide):

[0175]

[0176] or a salt thereof.

[0177] Suitably, (2,6-dichloro-4-(2-piperazin-1-yl-pyridin-4-yl)-N-(1,3,5-trimethyl-1H-pyrazol-4-yl)-benzenesulfonamide) or a salt thereof is attached to the linker via the NH group of the piperazinyl ring.

[0178] In one embodiment, the NMT inhibitor is 2,6-dichloro-N-(5-isobutyl-1,3-dimethyl-1H-pyrazol-4-yl-4-(-4-(2-piperazin-1-yl-pyridin-4-yl)-benzenesulfonamide:

[0179]

[0180] or a salt thereof.

[0181] Suitably, 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 attached to the linker via the NH group of the piperazinyl ring.

[0182] Compounds of formula (III), (IV), (IIIa), (2,6-dichloro-4-(2-piperazin-1-yl-pyridin-4-yl)-N-(1,3,5-trimethyl-1H-pyrazol-4-yl)-benzenesulfonamide) and 2,6-dichloro-N-(5-isobutyl-1,3-dimethyl-1H-pyrazol-4-yl-4-(-4-(2-piperazin-1-yl-pyridin-4-yl)-benzenesulfonamide are disclosed in WO2010 / 026365, the entire contents of which are incorporated herein by reference to describe the synthesis and activity of NMT inhibitors.

[0183] In one embodiment, the NMT inhibitor is a compound of formula (V):

[0184]

[0185] or a salt thereof,

[0186] in:

[0187] n1 is 1 or 2; n2 is 1 or 2;

[0188] X 1 Selected by CR x and N;

[0189] When present, R x is selected from the group consisting of hydrogen, halogen and -C optionally substituted with 1, 2 or 3 substituents. 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3;

[0190] R 1 is selected from the group consisting of: hydrogen; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OCH3 and -OCF3; and -C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from the group consisting of halogen, -CH3, -OCH3, and -OCF3;

[0191] R 2 is selected from the group consisting of: hydrogen; -C optionally substituted with 1, 2 or 3 substituents 1-4Alkyl, each substituent is independently selected from the group consisting of halogen, -OCH3 and -OCF3; and -C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from the group consisting of halogen, -CH3, -OCH3, and -OCF3;

[0192] or R 1 and R 2 are linked together so that, together with the atoms to which they are attached, they form C 3-6 A cycloalkyl group or a 3- to 6-membered non-aromatic heterocyclic group containing 1 heteroatom selected from the group consisting of O and N, wherein the C 3-6 The cycloalkyl or 3- to 6-membered non-aromatic heterocyclic group is optionally substituted with 1 or 2 substituents, each of which is independently selected from the group consisting of halogen, -OH, -CH3, -OCH3, and -OCF3;

[0193] R 3 is selected from the group consisting of: hydrogen; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; and -C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent independently selected from the group consisting of halogen, -OH, -CH3, -OCH3 and -OCF3;

[0194] or R 1 and R 3 are linked together so that together with the atoms to which they are attached they form a 3- to 6-membered non-aromatic heterocyclic group containing 1 N heteroatom, wherein the 3- to 6-membered non-aromatic heterocyclic group is optionally substituted with 1 or 2 substituents, each substituent being independently selected from the group consisting of halogen, -CH3, -OH, -OCH3, and -OCF3;

[0195] X 2 Selected by CR 4 and N;

[0196] When present, R 4 is selected from the group consisting of: hydrogen; halogen; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from halogen, -OH, -OCH3, -OCF3 and -NR a R b The group composed of;

[0197] R 5a and R 5dindependently selected from the group consisting of: hydrogen; halogen; methyl optionally substituted with 1, 2 or 3 substituents, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; and methoxy optionally substituted with 1, 2 or 3 substituents, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3;

[0198] R 5b and R 5c independently selected from the group consisting of: hydrogen; halogen; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; -OCF3 optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; and C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from the group consisting of halogen, -CH3, -OH, -OCH3, and -OCF3;

[0199] or R 5b and R 5c connected together so that they, together with the atoms to which they are connected, form a 6-membered aryl group or a 5- or 6-membered aromatic heterocyclic group comprising 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 heterocyclic group is optionally substituted with 1 or 2 substituents, each of which is independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3;

[0200] R 6 selected from the group consisting of hydrogen and methyl;

[0201] When present, each R 7 is -C optionally substituted by 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3;

[0202] R 8 is selected from the group consisting of: hydrogen; halogen; -OH; -CN; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OH, -CN and methoxy optionally substituted by 1, 2 or 3 halogens; -C 3-6Cycloalkyl, each substituent is independently selected from the group consisting of halogen, -CH3, -OH, -CN and methoxy optionally substituted by 1, 2 or 3 halogens; -C 1-4 alkenyl, each substituent independently selected from the group consisting of halogen, -OH, -CN, and methoxy optionally substituted with 1, 2 or 3 halogens; and -OC optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OH, CN, and methoxy optionally substituted with 1, 2 or 3 halogens;

[0203] R 9 is selected from hydrogen and -C optionally substituted by 1, 2 or 3 substituents 1-4 alkyl, each substituent being independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; or

[0204] R 8 and R 9 are linked together so that they, together with the atoms to which they are attached, form a 6-membered aromatic group, C 5-6 cycloalkyl, or a 5- or 6-membered aromatic heterocyclic group containing 1 or 2 heteroatoms selected from N, O and S, and wherein the 6-membered aromatic group, C 5-6 The cycloalkyl or 5- to 6-membered aromatic heterocyclic group is optionally substituted by 1, 2 or 3 substituents, each of which is independently selected from the group consisting of: halogen; -OH; -CN; -C optionally substituted by 1, 2 or 3 substituents. 1-4 alkyl, each substituent independently selected from the group consisting of halogen, -OH, and methoxy optionally substituted with 1, 2 or 3 halogens; and -OC optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of -halogen, -OH, and methoxy optionally substituted with 1, 2 or 3 halogens;

[0205] p is 0, 1, or 2;

[0206] Z is a 5- to 13-membered non-aromatic heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O and S, wherein at least one heteroatom is N, and wherein the 5- to 13-membered non-aromatic heterocyclic group is optionally substituted with 1, 2, 3 or 4 substituents, each of which is independently selected from the group consisting of: halogen; -C optionally substituted with 1, 2 or 3 substituents 1-6 Alkyl, each substituent is independently selected from halogen, -OH and -OC optionally substituted by 1, 2 or 3 halogens 1-3 alkyl; -OC optionally substituted by 1, 2 or 3 substituents 1-6Alkyl, each substituent is independently selected from halogen, -OH and -OC optionally substituted by 1, 2 or 3 halogens 1-3 Alkyl group; NR c R d and C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 or when two substituents are in adjacent ring positions, they may be linked together so that together with the atoms to which they are attached they form a C 3-6 Cycloalkyl or a 4- to 6-membered non-aromatic heterocyclic group containing 1 heteroatom selected from O and N, wherein the C 3-6 Cycloalkyl or 4 to 6 membered non-aromatic heterocyclic group is optionally substituted by 1 or 2 substituents, each substituent being independently selected from the group consisting of: halogen; -C 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 alkyl; and -OC optionally substituted by 1, 2 or 3 substituents 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 A group consisting of alkyl groups;

[0207] R c is hydrogen;

[0208] R d is selected from the group consisting of: hydrogen; -C optionally substituted with 1, 2 or 3 substituents 1-6 Alkyl, each substituent is independently selected from the group consisting of halogen, -OCH3 and -OCF3; and -C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from the group consisting of halogen, -CH3, -OH, -OCH3, and -OCF3;

[0209] or Z is -NR 10 R 11 ,in

[0210] R 10 is hydrogen; and

[0211] R 11is a 5- to 10-membered non-aromatic heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O and S, wherein at least one heteroatom is N, and wherein the 5- to 10-membered non-aromatic heterocyclic group is optionally substituted by 1, 2, 3 or 4 substituents independently selected from the group consisting of halogen; -OH; -C optionally substituted by 1, 2 or 3 substituents 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 alkyl; and -OC optionally substituted by 1, 2 or 3 substituents 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 alkyl; and

[0212] When present, each R a and R b are independently selected from hydrogen and -C 1-4 Alkyl group.

[0213] Suitably, the compound of formula (V) is linked to the linker via a nitrogen atom present in group Z, for example, via a nitrogen ring atom when group Z is a 5- to 13-membered non-aromatic heterocyclic group, or via NR present in group Z when group Z is a 5- to 13-membered non-aromatic heterocyclic group. c R d group, or through -NR 10 R 11 Connected to the linker. It will be understood by those skilled in the art that in order for the linker to be connected to the nitrogen atom present in the group Z (e.g., in the compounds of formula (V) and elsewhere herein), the nitrogen atom present in the group Z must be connected to at least one hydrogen atom, i.e., the nitrogen atom cannot be part of a tertiary amino group. In the conjugate formed between the compound (e.g., group Z) and the linker, the NH covalent bond in the group Z is replaced by the N-linker covalent bond.

[0214] In one embodiment, the NMT inhibitor is a compound of formula (Vq):

[0215]

[0216] or a salt thereof,

[0217] in:

[0218] R 3a is H or -C optionally substituted by 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OCH3 and -OCF3;

[0219] R 8ais halogen or C optionally substituted by 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OCH3 and -OCF3;

[0220] ra is 0, 1, or 2;

[0221] m a is 1 or 2;

[0222] R 12a is hydrogen; and

[0223] When present, each R 13a independently selected from the group consisting of: C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OCH3 and -OCF3; and C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent independently selected from the group consisting of halogen, -CH3, -OCH3 and -OCF3; or

[0224] When ra is 2 and two R 13a When the groups are located at adjacent ring positions, the two R 13a are linked together so that together with the atoms to which they are attached they form a C optionally substituted with 1 or 2 substituents. 3-6 Cycloalkyl, each substituent independently selected from the group consisting of: halogen; -C 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 Alkyl group.

[0225] Suitably, the compound of formula (Vq) or a salt thereof is prepared by NR 12a The group is attached to the linker.

[0226] 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"):

[0227]

[0228] or a salt thereof.

[0229] 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.

[0230] Compounds of formula (V), formula (Vq) and (S)-1-(5-chloro-2-(2-methylpiperazine-1-yl)pyrimidin-4-yl)-N-(2-(imidazo[1,2-a]pyridin-3-yl)propan-2-yl)azetidine-3-carboxamide are disclosed in WO2022 / 058745, the entire contents of which are incorporated herein by reference to describe the synthesis and activity of NMT inhibitors. Specifically, NMT inhibitor 26 can be prepared as described in Example 129 in WO2022 / 058745.

[0231] In one embodiment, the NMT inhibitor is a compound of formula (VI):

[0232]

[0233] or a salt thereof,

[0234] in:

[0235] R 1 is a group of formula OLA;

[0236] L is -(CHR 12 ) m -;

[0237] Each R 12 are independently H or C 1-4 alkyl;

[0238] m is 1, 2 or 3;

[0239] A is:

[0240]

[0241] v is 0, 1, or 2;

[0242] R 9a H, C 1-4 Alkyl or C 1-4 Haloalkyl;

[0243] R 9b H, C 1-4 Alkyl or C 1-4 Haloalkyl;

[0244] R 9c C 1-4 Alkyl or C 1-4 Haloalkyl;

[0245] R 9d H, C 1-4 Alkyl or C 1-4 Haloalkyl;

[0246] R 10 H, C 1-4 Alkyl or C 1-4 Haloalkyl;

[0247] R 11 H, halide, CN, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy or C 1-4 Haloalkoxy;

[0248] s is 0, 1, 2, or 3;

[0249] Each R 2 are independently F, Cl, Br, C optionally substituted with up to 3 halogen groups 1-4 Alkyl, OCH3 or OCF3;

[0250] Y is CH or C 1-4 alkyl;

[0251] R 3 H or C 1-4 alkyl;

[0252] R 4 H or C 1-4 alkyl;

[0253] R 5 is H;

[0254] R 6 H or C 1-4 alkyl;

[0255] q is 0 or 1;

[0256] R 7 is H or methyl;

[0257] R 8 is H or methyl;

[0258] or R 3 and R 6 and the atoms therebetween form bonds between the atoms therebetween, or between the atoms therebetween and -(CHR a ) r - a 3- to 7-membered non-aromatic heterocyclic ring; or R 7 Group and R 6 The groups and the atoms between them form the atoms between them and -(CHR a) r -a 3- to 7-membered non-aromatic heterocyclic ring;

[0259] r is 1, 2, 3, 4 or 5; and

[0260] R a It is hydrogen or methyl.

[0261] Suitably, the compound of formula (VI) or a salt thereof is 5 )(R 6 ) group is connected to the linker. In one embodiment, the NMT inhibitor is a compound of formula (VIA):

[0262]

[0263] or a salt thereof,

[0264] in:

[0265] R 2a is H or F;

[0266] R 2b is F;

[0267] R 5a is H;

[0268] R 6a is H or methyl;

[0269] R 9ca is methyl, isopropyl or tert-butyl;

[0270] R 9cb is H or methyl;

[0271] R 10a is methyl; and

[0272] R 11a is methyl;

[0273] The condition is that when R 2a When H, R 9cb For H.

[0274] Suitably, the compound of formula (VIA) or a salt thereof is 5a )(R 6a ) group is attached to the linker.

[0275] 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:

[0276]

[0277] or a salt thereof.

[0278] Suitably, 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 linked to the linker via an NH(Me) group.

[0279] 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:

[0280]

[0281] or a salt thereof.

[0282] Suitably, 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 linked to the linker via a NH(Me) group.

[0283] Compounds of formula (VI), formula (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 European Patent Application No. 22194959.7, the entire contents of which are incorporated herein by reference to describe the synthesis and activity of NMT inhibitors. The compounds can also be prepared according to methods known to those skilled in the art, such as the methods disclosed herein for synthesizing NMT inhibitor 1.

[0284] In one embodiment of the 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, a NMT inhibitor is provided.

[0285] Linker

[0286] 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 connect one or more NMT inhibitors and antibodies to form the ADC of the present invention. The linker can be outside the cell, i.e., extracellular, stable, or can be cleaved by enzyme activity, hydrolysis or other metabolic conditions. In one embodiment, the linker is a cleavable linker, such as a linker that can be cleaved by an enzyme (particularly a lysosomal enzyme).

[0287] The ADC of the present invention can be conveniently prepared using a linker with a reactive functional group that binds to the NMT inhibitor and the antibody. A cysteine ​​thiol or amine of the antibody, such as a chain terminus (e.g., N-terminus), or an amino acid side chain (e.g., lysine), can form a bond with a functional group of the linker (e.g., maleimide).

[0288] The linker is preferably stable outside the cell. Before transport or delivery to the cell, the ADC of the present invention is preferably stable and remains intact, i.e., the antibody remains connected to the NMT inhibitor. The linker is stable outside the target cell and can be cleaved at a certain effective rate inside the cell. An effective linker will: (i) maintain the specific binding properties of the antibody; (ii) allow the conjugate or drug moiety to be delivered inside the cell; (iii) remain stable and intact, i.e., not cleaved, until the conjugate is delivered or transported to its target site; and (iv) once cleaved, maintain the cytotoxicity, cell killing or cell inhibition of the NMT inhibitor. The stability of the ADC can be measured by standard analytical techniques, such as mass spectrometry, HPLC, and separation / analysis techniques LC / MS. Derivatization studies (data not shown) lead the applicant to conclude that the ADC of the present invention does not have NMT inhibitory activity, and therefore the NMT inhibitory activity is achieved by cleaving the linker to release the NMT inhibitor.

[0289] Covalent attachment of the antibody and drug moieties requires that the linker has two reactive functional groups, i.e., divalency in the sense of reaction. Bivalent linker reagents are known to be useful for attaching two or more functional or biologically active moieties, such as peptides, nucleic acids, drugs, toxins, antibodies, haptens, and reporter groups, and methods for producing conjugates thereof have been described (Hermanson, GT (1996) Bioconjugate Techniques; Academic Press: New York, pp. 234-242).

[0290] The linker can be substituted with groups that modulate cohesion, solubility, or reactivity. For example, a sulfonate substituent can increase the water solubility of the reagent and facilitate the coupling reaction of the linker reagent to the antibody or drug moiety, or to the linker and NMT inhibitor to the antibody or to the linker and antibody to the NMT inhibitor, depending on the synthetic route used to prepare the ADC.

[0291] In one embodiment, the linker unit has formula (VII):

[0292] -A a -W w -Y y -(Formula (VII))

[0293] in:

[0294] A is a first stretcher unit which, when present, forms a covalent bond with a chain terminus (e.g., N-terminus) or a functional group of an amino acid side chain of an antibody;

[0295] a is 0 or 1;

[0296] Each W is independently an amino acid unit or a glucuronide unit, which, when A and / or Y are absent, forms a covalent bond with a chain terminus (e.g., N-terminus) or a functional group of an amino acid side chain of an antibody and / or a functional group of an NMT inhibitor, respectively;

[0297] When W is an amino acid unit, w is 1 to 12;

[0298] When W is a glucuronide unit, w is 1 or 2;

[0299] Y is a second stretcher unit which, when present, forms a covalent bond with the functional group of the NMT inhibitor; and

[0300] y is either 0 or 1.

[0301] The NMT inhibitor forms a covalent bond with the second stretcher unit (Y) (when present), or when Y is absent, forms a covalent bond with the amino acid unit or the glucuronide unit (W). Suitably, the functional group on the NMT inhibitor is an amino group or an alcohol, such as an amino group. When the NMT inhibitor forms a covalent bond through an amino group, the amino group must have an available hydrogen atom to react with the corresponding functional group (e.g., carbonyl) in the linker, i.e., the amino group cannot be a tertiary amino group.

[0302] The first extension unit

[0303] When present, the extension unit (A) is capable of connecting the antibody to an amino acid unit or a glucuronide unit (W). In this regard, the antibody has a functional group that can form a key with the functional group of the extension unit, such as a functional group of the amino acid side chain of the antibody. Useful functional groups that may exist on the antibody (whether natural or by chemical manipulation) include but are not limited to sulfhydryl (-SH), amino, hydroxyl, carboxyl, anomeric hydroxyl and carboxyl of a carbohydrate. In some embodiments, the antibody functional group is a sulfhydryl and / or amino group, especially a sulfhydryl. The sulfhydryl can be produced by reducing the intramolecular disulfide bonds of the antibody. The sulfhydryl can also be generated by the reaction of the amino group of the antibody lysine moiety using 2-iminothiolane (Traut reagent) or another sulfhydryl generating reagent.

[0304] In one embodiment, the first stretcher forms a bond with a sulfur atom of the antibody. The sulfur atom may be derived from a thiol group of the antibody.

[0305] In one embodiment, the stretcher unit (A) has the formula (A1):

[0306]

[0307] in:

[0308] n is 1 to 6;

[0309] indicates the point of attachment to the chain terminus (e.g., N-terminus) or a functional group of an amino acid side chain of an antibody; Indicates the connection point with W.

[0310] In one embodiment, A is (A2):

[0311]

[0312] In one embodiment, A is (A3):

[0313]

[0314] Amino Acid and Glucuronide Units

[0315] Each W is independently an amino acid unit or a glucuronide unit, which, when A and / or Y are absent, forms a covalent bond with the chain terminus (eg, N-terminus) or a functional group of an amino acid side chain of the antibody and / or a functional group of the NMT inhibitor, respectively.

[0316] In some embodiments, each W is independently an amino acid unit. In such embodiments, w is 1 to 12, such as 1, 2 or 3, especially 2.

[0317] In some embodiments, the amino acid unit can be cleaved by one or more enzymes, such as tumor-associated proteases, thereby promoting release of the NMT inhibitor from the ADC upon exposure to intracellular proteases, such as lysosomal enzymes (Doronina et al. (2003) Nat. Biotechnol. 21:778-784).

[0318] In one embodiment, each W has the formula (WI):

[0319]

[0320] where w is as defined above; and

[0321] R 19 is 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,

[0322]

[0323] in represents the point of attachment to A (when present) or to a chain terminus (e.g., the N-terminus) or an amino acid side chain functional group of the antibody; and

[0324] represents the point of attachment to Y (when present) or to a functional group of the NMT inhibitor.

[0325] In one embodiment, at least one R 19 In one embodiment, at least one R 19 It is (CH2)3NHC(=NH)NH2.

[0326] In one embodiment, (W)w has the formula (WII):

[0327]

[0328] in:

[0329] R 19a is H, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, CH2OH, CH(OH)Me, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, phenyl,

[0330]

[0331] and

[0332] R 19b It 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, (C H2)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.

[0333] In one embodiment, R 19a In one embodiment, R 19b It is (CH2)3NHC(=NH)NH2.

[0334] In one embodiment, each W is a glucuronide unit. In such embodiments, w is 1 or 2, especially 1. The glucuronide unit includes a site cleavable by β-glucuronidase.

[0335] In one embodiment, W (particularly (W)w) has the formula (WIII):

[0336]

[0337] in:

[0338] R is H, halide, CN or NO2;

[0339] m is 0, 1, 2 or 3;

[0340] Su is the sugar part;

[0341] in represents the point of attachment to A (when present) or to a chain terminus (e.g., the N-terminus) or a functional group of an amino acid side chain of the antibody; and

[0342] represents the point of attachment to Y (when present) or to a functional group of the NMT inhibitor.

[0343] 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 NO2.

[0344] 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.

[0345] In one embodiment, the sugar moiety (Su) is a cyclic hexose, such as pyranose, or a cyclic pentose, such as furanose. In one embodiment, the pyranose is a glucuronide or a hexose. Suitably, the sugar moiety is in a β-D conformation. In one embodiment, the pyranose is a β-D-glucuronide moiety (i.e., β-D-glucuronic acid is connected to the remainder of W by a glycosidic bond that can be cleaved 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., substituted by one or more groups (e.g., hydrogen, hydroxyl, halogen, sulfur, nitrogen or C 1-6 alkyl) substituted glucuronic acid).

[0346] In one embodiment, Su has the formula:

[0347]

[0348] The second extension unit

[0349] When present, the second stretching unit (Y) connects W to the NMT inhibitor by forming a covalent bond with a functional group of the NMT inhibitor. For example, Y may comprise 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 react with the corresponding functional group (e.g., carbonyl) of Y, i.e., the amino group cannot be a tertiary amino group. The second stretching unit may be a "self-destructive" or "non-self-destructive" group.

[0350] A "non-self-destructive" stretch unit means that after the ADC is enzymatically (e.g., proteolytically) cleaved, some or all of the stretch unit remains bound to the drug moiety. Examples of non-self-destructive spacer units include, but are not limited to, glycine stretch units and glycine-glycine stretch units. Other combinations of peptide stretch units that are susceptible to sequence-specific enzymatic cleavage are also contemplated. For example, enzymatic cleavage of an ADC containing a glycine-glycine stretch unit by a tumor cell-associated protease will result 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 then undergoes a hydrolysis step in the tumor cell, thereby cleaving the glycine-glycine spacer unit from the NMT inhibitor moiety.

[0351] The “self-destructive” spacer unit allows the release of the NMT inhibitor without the need for a separate hydrolysis step.

[0352] When W is an amino acid unit, y is suitably 1. When W is a glucuronide unit, y is suitably 0.

[0353] In one embodiment, the second stretching unit (Y) has the formula (Y1):

[0354]

[0355] in:

[0356] Each Q is independently a halide, NO2, CN, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 Haloalkoxy;

[0357] m is 0 to 4;

[0358] in represents the point of connection with W; and

[0359] represents the point of attachment to the functional group of the NMT inhibitor.

[0360] In one embodiment, m is not zero and at least one Q is halo. In a second embodiment, m is not zero and at least one Q is NO2. In a third embodiment, m is not zero and at least one Q is CN. In a fourth embodiment, m is not zero and at least one Q is C 1-6 In a fifth embodiment, m is not zero and at least one Q is C 1-6 In a sixth embodiment, m is not zero and at least one Q is C 1-6 In a seventh embodiment, m is not zero and at least one Q is C1-6 Halogenated alkoxy.

[0361] 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.

[0362] Suitable examples of self-destructive stretching units also include, but are not limited to, aromatic compounds electronically similar to p-aminobenzyl alcohol and its derivatives (e.g., formula (Y1) (see, e.g., US2005 / 0256030 Al)), such as 2-aminoimidazole-5-methanol derivatives (Hay et al. (1999) Bioorg. Med. Chem. Lett. 9:2237) and o-aminobenzyl acetal or p-aminobenzyl acetal. Spacers undergo cyclization after amide bond hydrolysis, such as substituted and unsubstituted 4-aminobutyric acid amide (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 (Amsberry 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 useful self-immolative stretching unit in ADCs.

[0363] In one embodiment, the linker has formula (LI):

[0364]

[0365] in represents the point of attachment to a functional group on the chain terminus (e.g., N-terminus) or amino acid side chain of the antibody; and

[0366] represents the point of attachment to the functional group of the NMT inhibitor.

[0367] In one embodiment, the linker has formula (LII):

[0368]

[0369] in represents the point of attachment to a functional group on the chain terminus (e.g., N-terminus) or amino acid side chain of the antibody; and

[0370] represents the point of attachment to the functional group of the NMT inhibitor.

[0371] In one embodiment, the linker has formula (LIII):

[0372]

[0373] in represents the point of attachment to a functional group on the chain terminus (e.g., N-terminus) or amino acid side chain of the antibody; and

[0374] represents the point of attachment to the functional group of the NMT inhibitor.

[0375] In one embodiment, the linker has formula (LIV):

[0376]

[0377] in represents the point of attachment to a functional group on the chain terminus (e.g., N-terminus) or amino acid side chain of the antibody; and

[0378] represents the point of attachment to the functional group of the NMT inhibitor.

[0379] Antibody

[0380] The term "antibody" as used herein refers to monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), complete antibodies, and antibody fragments, as long as they exhibit the desired biological activity. Antibodies can 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 edition, Garland Publishing, New York). Target antigens typically have many binding sites, also referred to as epitopes, recognized by CDRs on a variety of antibodies. Each antibody that specifically binds to a different epitope has a different structure. Therefore, an antigen may have more than one corresponding antibody. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules that include antigen binding sites or portions thereof that immunospecifically bind to target antigens of interest, such targets include but are not limited to cancer cells or cells that produce autoimmune antibodies associated with autoimmune diseases. The immunoglobulin can be an immunoglobulin molecule of any type (e.g., IgG, IgE, IgM, IgD, and IgA), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass, or any 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). The immunoglobulin can be from any species, including human, mouse, or rabbit origin.

[0381] "Antibody fragments" include a portion of a full-length antibody, typically its antigen binding or variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, and scFv fragments; diabodies; linear antibodies; fragments produced by Fab expression libraries, anti-idiotypic (anti-ld) antibodies, CDRs (complementarity determining regions), and epitope binding fragments of any of the above that immunospecifically bind to cancer cell antigens, viral antigens, or microbial antigens, single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0382] The term "monoclonal antibody" as used herein refers to an antibody obtained from a substantially homogeneous antibody population, that is, each antibody comprising the population is identical, except for possible naturally occurring mutations that may be present in trace amounts. Monoclonal antibodies have a high degree of specificity (for a single antigenic site). In addition, in contrast to polyclonal antibody preparations comprising different antibodies for different determinants (epitopes), each monoclonal antibody is for a single determinant on an antigen. In addition to its specificity, the advantage of monoclonal antibodies is that they can be synthesized but not contaminated by other antibodies. The modifier "monoclonal" indicates the characteristics of the antibody obtained from a substantially homogeneous antibody population and should not be construed as requiring the antibody to be produced by any ad hoc method. For example, the monoclonal antibody used according to the present invention can be prepared by the hybridoma method first described by the people such as Kohler (1975) Nature 256:495, or can be prepared by a recombinant DNA method (see US 4816567). Monoclonal antibodies can also be isolated from phage antibody libraries using the techniques described by Clackson et al. (1991) Nature, 352:624-628; Marks et al. (1991) J. Mol. Biol., 222:581-597, or from transgenic mice carrying a complete human immunoglobulin system (Lonberg (2008) Curr. Opinion 20(4):450-459).

[0383] The monoclonal antibodies herein specifically include "chimeric" antibodies, in which a portion of the heavy chain and / or light chain is identical or homologous to the corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, and the remainder of the chain is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, as long as they exhibit the desired biological activity (US 4816567; and Morrison et al. (1984) Proc. Natl. Acad. Sci. USA, 81: 6851-6855). Chimeric antibodies include "primatized" antibodies, which include variable domain antigen binding sequences derived from non-human primates (e.g., old world monkeys or apes) and human constant region sequences.

[0384] "Complete antibodies" herein are antibodies comprising VL and VH domains, 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. A complete antibody may have one or more "effector functions," which refer to those biological activities attributable to the Fc region of an antibody (a native sequence Fc region or an amino acid sequence variant Fc region). Examples of antibody effector functions include C1q binding; complement dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis and downregulation of cell surface receptors such as B cell receptors and BCRs.

[0385] Depending on the amino acid sequence of its heavy chain constant domain, whole antibodies can be assigned to different "classes". There are five major classes of whole 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 corresponding to different antibody classes are called α, δ, ε, γ and μ, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[0386] The desired biological activity of the antibody suitably comprises binding to a cancer associated antigen. The binding is suitably specific, i.e. the antibody preferentially binds to the cancer associated antigen and does not significantly bind to other antigens or proteins (eg BSA).

[0387] 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 hematologic cancer cell, such as a B cell or a T cell. Therefore, antibodies directed against antigens expressed by cancer cells bind to the cancer cells and (without limitation by theory) mediate cell death through the action of NMT inhibitors and / or antibody-mediated cytotoxicity (ADCC). Cancer-associated antigens may sometimes be expressed by non-cancerous cells, but are preferably preferentially expressed by cancer cells. An example of a cancer-associated antigen is HER2. HER2 is particularly expressed on certain breast cancer cells. Another example of a cancer-associated antigen is CD20. CD20 is expressed on certain B cells, including certain B cell lymphoma cells.

[0388] As used herein, "binding to HER2" is used to indicate that the binding affinity of the antibody to HER2 is higher than that of a non-specific partner, such as bovine serum albumin (BSA, Genbank accession number CAA76847, version number CAA76847.1 GI: 3336842, record update date: January 7, 2011 at 02:30 p.m.). In some embodiments, when measured under physiological conditions, the association constant (Ka) of the antibody binding to HER2 is at least 2, 3, 4, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, 104, 105 or 106 times the association constant of the antibody to BSA. The antibodies of the present invention can bind to HER2 with high affinity. For example, in some embodiments, the antibodies can bind to HER2 with an affinity equal to or less than about 10 -6 M (e.g. 1x 10 -6 , 10 -7 , 10 -8 , 10 -9 , 10 -10 , 10 -11 , 10 -12 , 10 -13 or 10 -14 ) with a KD of 1.5 Å to HER2.

[0389] 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. AAA75493.1GI:306840, Record Update Date: June 23, 2010 08:47 AM. In one embodiment, the nucleic acid encoding the HER2 polypeptide corresponds to Genbank Accession No. M11730, Version No. M11730.1GL183986, Record Update Date: June 23, 2010 08:47 AM.

[0390] Antibody modification

[0391] The antibodies disclosed herein can be modified, for example, to make them less immunogenic to human subjects. This can be achieved by a variety of techniques familiar to those skilled in the art. Some of these techniques will be described in more detail below.

[0392] Humanization

[0393] The technology of reducing the immunogenicity of non-human antibodies or antibody fragments in vivo includes so-called "humanization" technology. "Humanized antibody" refers to a polypeptide of at least a portion of the variable region comprising the modification of human antibodies, wherein a portion of the variable region (preferably, significantly less than the portion of the complete human variable domain) has been replaced by the corresponding sequence from non-human species, and wherein the variable region modified is connected to at least another portion of another protein (preferably, the constant region of human antibodies). The statement "humanized antibody" includes human antibody source antibodies, wherein one or more complementary determining regions (" CDR ") amino acid residues and / or one or more framework regions (" FW " or " FR ") amino acid residues are replaced by amino acid residues from rodents or other non-human similar sites. The statement "humanized antibody" also includes immunoglobulin amino acid sequence variants or fragments thereof, which include FRs substantially having human immunoglobulin amino acid sequences and CDRs substantially having non-human immunoglobulin amino acid sequences.

[0394] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies containing minimal sequences derived from non-human immunoglobulins. Alternatively, from another perspective, a humanized antibody is a human antibody that also contains selected sequences from non-human (e.g., murine) antibodies to replace human sequences. Humanized antibodies may include conservative amino acid substitutions or non-natural residues from the same or different species, but will not significantly change their binding and / or biological activity. Such antibodies are chimeric antibodies containing minimal sequences from non-human immunoglobulins.

[0395] There are many humanization techniques, including "CDR grafting", "guide selection", "deimmunization", "surface reshaping" (also known as "veneer"), "complex antibodies", "human string content optimization" and framework shuffling.

[0396] CDR transplantation

[0397] In this technique, humanized antibodies are human immunoglobulins (recipient antibodies) in which residues in the complementary determining regions (CDRs) of the recipient antibody are replaced by residues from CDRs of a non-human species (donor antibody) (e.g., mouse, rat, camel, cow, goat, or rabbit) with the desired properties (in effect, the non-human CDRs are "grafted" onto a human framework). In some cases, framework region (FR) residues of human immunoglobulins are replaced by corresponding non-human residues (for example, when a FR residue has a significant effect on antigen binding).

[0398] In addition, humanized antibodies can include residues that are neither present in the receptor antibody nor in the CDR or framework sequences of the input. These modifications are made in order to further improve and maximize antibody performance. Therefore, usually humanized antibodies will include at least one and all of two variable domains on the one hand, wherein all or all of the hypervariable loops correspond to those hypervariable loops of non-human immunoglobulins and all or substantially all of the FR regions are those FR regions of human immunoglobulin sequences. The humanized antibodies will optionally also include at least a portion of an immunoglobulin constant region (Fc), or at least a portion of an immunoglobulin constant region (Fc) of a human immunoglobulin.

[0399] Guide selection

[0400] The method consists of combining the VH or VL domain of a given non-human antibody specific for a specific epitope with a human VH or VL library, and selecting a specific human V domain for the target antigen. The selected human VH is then combined with the VL library to generate a fully human VHxVL combination. The method is described in Nature Biotechnology (NY) 12, (1994) 899-903.

[0401] Compound Antibody

[0402] In this method, two or more amino acid sequence fragments of human antibodies are combined in the final antibody molecule. They are constructed by combining multiple human VH and VL sequence fragments, and the combination limits or avoids human T cell epitopes in the final composite antibody V region. When necessary, T cell epitopes can be limited or avoided by exchanging the V region fragments that will contribute to or encode T cell epitopes with alternative fragments that avoid T cell epitopes. This method is described in US2008 / 0206239A1.

[0403] Deimmunization

[0404] This method involves removing T cell epitopes of people (or other second species) from the V region of therapeutic antibodies (or other molecules). By comparing, for example, with an MHC binding motif database (e.g., a "motif" database hosted on www.wehi.edu.au), the presence of MHC class II binding motifs in the V region sequence of therapeutic antibodies is analyzed. Alternatively, computational threading methods (such as those designed by Altuvia et al. (J.Mol.Biol.249 244-250 (1995)) can be used to identify MHC class II binding motifs; in these methods, continuous overlapping peptides from V region sequences are tested for binding energy to MHC class II proteins. These data can then be combined with information on other sequence features of the successfully presented peptide, such as amphipathicity, Rothbard motifs, and cleavage sites of protease B and other processing enzymes.

[0405] Once a potential second species (e.g., human) T cell epitope has been identified, it is eliminated by changing 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 depending on the primary or secondary structure of the protein (and therefore, may not be adjacent in the primary structure). The most typical change is by substitution, but in some cases, the addition or deletion of amino acids may be more appropriate.

[0406] All changes can be made by recombinant DNA technology, so the final molecule can be prepared by expression from a recombinant host using well-established methods such as site-directed mutagenesis. However, it is also possible to use protein chemistry or any other method of molecular alteration.

[0407] Surface reshaping

[0408] This method includes:

[0409] (a) determining the conformational structure of a variable region of a non-human (e.g., rodent) antibody (or fragment thereof) by constructing a three-dimensional model of the variable region of the non-human antibody;

[0410] (b) using the relative accessibility distribution, generating a sequence alignment from a sufficient number of X-ray crystal structures of non-human and human antibody variable region heavy and light chains to provide a set of heavy and light chain framework positions, wherein the aligned positions are 98% identical in the sufficient number of non-human antibody heavy and light chains;

[0411] (c) defining a set of heavy chain and light chain surface exposed amino acid residues for the non-human antibody to be humanized using the set of framework positions generated in step (b);

[0412] (d) identifying a group of heavy chain and light chain surface exposed amino acid residues that are most identical to the surface exposed amino acid residues defined in step (c) from a human antibody amino acid sequence, wherein the heavy chain and light chain from the human antibody are or are not naturally paired;

[0413] (e) replacing the heavy chain and light chain surface exposed amino acid residue groups defined in step (c) with the heavy chain and light chain surface exposed amino acid residue groups identified in step (d) in the amino acid sequence of the non-human antibody to be humanized;

[0414] (f) constructing a three-dimensional model of the variable region of the non-human antibody produced by the substitutions specified in step (e);

[0415] (g) identifying, by comparing the three-dimensional models constructed in steps (a) and (f), any amino acid residue in 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

[0416] (h) changing any residue identified in step (g) from a human amino acid residue to an original non-human amino acid residue, thereby defining a set of surface exposed amino acid residues for humanization of the non-human antibody; provided that step (a) does not have to be performed first, but must be performed before step (g).

[0417] Superhumanization

[0418] This method compares the non-human sequence with a functional human germline gene library. Human genes that encode the same or closely related typical structures as the non-human sequence are selected. Human genes with the highest homology within the CDR are selected as FR donors. Finally, the non-human CDRs are transplanted onto these human FRs. This method is described in patent WO 2005 / 079479 A2.

[0419] Human Presenting Peptide Optimization

[0420] This method compares non-human (e.g., mouse) sequences to the human germline gene library and scores the differences as human presented peptides (HSC), thereby quantifying the sequence at the level of potential MHC / T cell epitopes. Multiple different humanized variants are then generated by maximizing their HSC rather than using a global identity measure (described in Molecular Immunology, 44, (2007) 1986-1998), thereby humanizing the target sequence.

[0421] Framework Reshuffle

[0422] The CDRs of the non-human antibody are fused in-frame to a cDNA pool containing all known heavy and light chain human germline gene frameworks. Humanized antibodies are then selected, for example, by screening a phage display antibody library. This is described in Methods 36, 43-60 (2005).

[0423] In one embodiment, antibodies known to be used for the treatment or prevention of cancer are used according to 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 used to treat patients with metastatic breast cancer (Stebbing, J., Copson, E. and O'Reilly, S. "Herceptin (trastuzamab) 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 used to treat 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 Inc., NY), a murine IgG antibody for the treatment of lung cancer; IMC-C225 (Imclone Systems Inc., NY), a chimeric IgG antibody for the treatment of head and neck cancer; Vitaxin (Medhnmune, Inc., MD), a humanized antibody for the treatment of sarcoma; Campath 1 / H (Leukosite, MA), a humanized IgGi antibody for the treatment of chronic lymphocytic leukemia (CLL); Smart MI95 (Protein Design Labs, Inc., CA), a humanized IgG antibody for the treatment of acute myeloid leukemia (AML); LymphoCide (Immunomedics, Inc., NJ), a humanized IgG antibody for the treatment of non-Hodgkin's lymphoma; Smart ID 10 (Protein Design Labs, Inc., CA), a humanized antibody for the treatment of non-Hodgkin's lymphoma; Oncolym (Techniclone, Inc., CA), a mouse antibody for the treatment of non-Hodgkin's lymphoma; Allomune (BioTransplant, CA), a humanized anti-CD2 mAb for the treatment of Hodgkin's disease or non-Hodgkin's lymphoma; anti-VEGF (Genentech, Inc., CA), a humanized antibody for the treatment of lung cancer 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 used to treat colorectal cancer, lung cancer and melanoma; cetuximab (ImClone, NJ), an anti-EGFR chimeric antibody used to treat epidermal growth factor-positive cancers. More examples of antibodies that bind to CD20 include ocrelizumab, obinutuzumab, ofatumumab, ibritumomab tiuxetan, tositumomab and ublituximab.

[0424] Other antibodies that can be used to treat cancer include, but are not limited to, antibodies against the following cancer-associated antigens: CA125 (ovarian), CA15-3 (cancer), CAI9-9 (cancer), L6 (cancer), Lewis Y (cancer), Lewis X (cancer), alpha-fetoprotein (cancer), CA 242 (colorectal), placental alkaline phosphatase (cancer), prostate-specific antigen (prostate), prostatic acid phosphatase (prostate), epidermal growth factor (cancer), MAGE-1 (cancer), MAGE-2 (cancer), MAGE-3 (cancer), MAGE-4 (cancer), anti-transferrin receptor (cancer), p97 (melanoma), MUC1-KLH (breast cancer), CEA (colorectal), gplOO (melanoma), MARTI (melanoma), 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 (cancer), CD38 (multiple myeloma), CD40 (lymphoma), mucin (cancer), P21 (cancer), MPG (melanoma), and Neu oncogene products (cancer). Some specific useful antibodies include, but are not limited to, BR96 mAb (Trail, PA, Willner, D., Lasch, SJ, Henderson, AJ, Hofstead, SJ, Casazza, AM, Firestone, RA, Hellstrom, I., Hellstrom, KE, "Cure of Xenografited 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, Casaz Za, AM, Braslawsky, GR, Brown, JP, 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 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-CD-40 antibody, SGN-14" Cancer Res. 2000, 60, 3225-3231), mAbs against CD70 antigen, such as 1F6 mAb, and mAbs against 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 also 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: acoming of age" Semin Oncol. 2000, 27, 64-70; Breitling, F., and Dubel, S., Recombinant Antibodies, John Wiley, and Sons, New York, 1998).

[0425] Other antibodies that can be used to treat 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 herein by reference for the purpose of defining antibodies.

[0426] In one embodiment, the antibody is selected from the group consisting of gemtuzumab, brentuximab, ado-trastuzumab, fam-trastuzumab, inotuzumab, polatuzumab, acituzumab, sacituzumab, belantamab, ioncastuximab, tisotumab, indatuximab, naratuximab, and depatuxizumab.

[0427] In another embodiment, the antibody is taciturnumab. Alternatively, the antibody is ifenatumomab.

[0428] Suitably, the antibody is selected from the group consisting of trastuzumab, rituximab, taciturnumab and ifenatumomab.

[0429] In one embodiment, the antibody is a humanized antibody, a chimeric antibody, a human antibody, or an antibody fragment.

[0430] In one embodiment, the antibody is combined with 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. Suitably, the antibody is trastuzumab. In one embodiment, the antibody is an antibody with 6 CDRs of trastuzumab. Trastuzumab comprises a heavy chain of SEQ ID NO:2 and a light chain of SEQ ID NO:1.

[0431] In one embodiment, the antibody binds to CD20. Suitably, the antibody is rituximab. In one embodiment, the antibody is an antibody having 6 CDRs of rituximab. Rituximab comprises a heavy chain of SEQ ID NO: 4 and a light chain of SEQ ID NO: 3.

[0432] 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.

[0433] In one embodiment, the antibody binds to a member of Claudin 18, in particular to Claudin 18.2, and, for example, an ADC comprising the antibody can be used to treat gastric cancer. In one embodiment, the cancer expresses one or more members of Claudin 18, in particular Claudin 18.2.

[0434] In one embodiment, the antibody binds to the folate receptor, and, for example, an ADC comprising the antibody can be used to treat ovarian cancer and endometrial cancer. In this embodiment, a suitable antibody is mirvetuximab. In one embodiment, the cancer expresses the folate receptor.

[0435] 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 can be used to treat various types of epithelial cell cancers. In one embodiment, the cancer expresses an EGF receptor.

[0436] In one embodiment, the antibody binds to Nectin 4, and, for example, an ADC comprising the antibody can be used to treat urothelial carcinoma, bladder cancer, pancreatic cancer, triple-negative breast cancer, non-small cell lung cancer, gastric cancer, esophageal cancer, or ovarian cancer. In this embodiment, suitably, the antibody is enfortumab. In one embodiment, the cancer expresses Nectin 4.

[0437] In one embodiment, the antibody binds to CD22, and, for example, an ADC comprising the antibody can be used to treat non-Hodgkin lymphoma. In one embodiment, the cancer expresses CD22.

[0438] In one embodiment, the antibody binds to c-MET, and, for example, an ADC comprising the antibody can be used to treat non-small cell lung cancer (NSCLC). In one embodiment, the cancer expresses c-MET.

[0439] In one embodiment, the antibody binds to Trop-2, and, for example, an ADC comprising the antibody can be used to treat metastatic triple-negative breast cancer and metastatic urothelial carcinoma. In this embodiment, a suitable antibody is taciturnin. In one embodiment, the cancer expresses Trop-2. Taciturnin comprises a heavy chain of SEQ ID NO: 8 and a light chain of SEQ ID NO: 7.

[0440] In one embodiment, the antibody binds to tissue factor, and, for example, an ADC comprising the antibody can be used to treat previously treated recurrent or metastatic cervical cancer. In this embodiment, suitably, the antibody is tisomonab. In one embodiment, the cancer expresses tissue factor.

[0441] In one embodiment, the antibody binds to CD276 (B7-H3), and, for example, an ADC comprising the antibody can be used to treat prostate cancer. In this embodiment, suitably, the antibody is ifenatumab. In one embodiment, the cancer expresses CD276 (B7-H3). Ifenatumab comprises a heavy chain of SEQ ID NO: 6 and a light chain of SEQ ID NO: 5.

[0442] In one embodiment, the antibody binds to receptor tyrosine kinase-like orphan receptor 1 (ROR1), and, for example, ADCs comprising the antibody can be used to treat various hematological and solid malignancies. In one embodiment, the cancer expresses tyrosine kinase-like orphan receptor 1 (ROR1).

[0443] In one embodiment, the antibody binds to BCMA, and, for example, an ADC comprising the antibody can be used to treat multiple myeloma. In one embodiment, the cancer expresses BCMA.

[0444] 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.

[0445] In one embodiment, the antibody binds to CEACAM5, and, for example, an ADC comprising the antibody can be used to treat advanced non-small cell lung cancer (NSCLC). In one embodiment, the cancer expresses CEACAM5.

[0446] In one embodiment, the antibody binds to mesothelin, and, for example, an ADC comprising the antibody can be used to treat mesothelioma, ovarian cancer, pancreatic cancer, gastric cancer, and non-small cell lung tumors. In one embodiment, the cancer expresses mesothelin.

[0447] In one embodiment, the antibody binds to AXL, and, for example, ADCs comprising the antibody can be used to treat a variety of solid tumors, including non-small cell lung cancer (NSCLC) and triple negative breast cancer (TNBC). In this embodiment, suitably, the antibody is enapotamab. In one embodiment, the cancer expresses AXL.

[0448] Drug loading

[0449] The drug loading (referred to as the variable "p") is the average number of NMT inhibitors per antibody. When the compounds of the invention are conjugated to cysteine ​​residues, the drug loading may range from 1 to 10 NMT inhibitors per antibody, i.e., wherein 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 NMT inhibitors are covalently attached to the antibody. The composition of the conjugate includes a collection of antibodies conjugated to 1 to 10 NMT inhibitors.

[0450] In the process of preparing ADC by coupling reaction, the average number of NMT inhibitors corresponding to each antibody can be characterized by conventional methods, such as UV, reverse phase HPLC, HIC, mass spectrometry, ELISA assay and electrophoresis. The quantitative distribution of ADC can also be determined according to p. By ELISA, the average value of p in a specific ADC preparation can be determined (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 limitations of antibody-antigen binding and ELISA, the distribution of p (NMT inhibitor) values ​​cannot be distinguished. In addition, the ELISA assay used to detect ADC cannot determine the connection position of the drug part to the antibody, such as heavy chain or light chain fragments, or specific amino acid residues. In some cases, the separation, purification and characterization of uniform ADCs (where p is a certain value of ADCs with other NMT inhibitor drug loadings) can be achieved by methods such as reverse phase HPLC or electrophoresis. These techniques are also applicable to other types of conjugates.

[0451] For some ADCs, p may be limited by the number of attachment sites on the antibody. For example, an antibody may have only one or a few cysteine ​​thiol groups, or may have only one or a few sufficiently reactive thiol groups through which a linker may be attached. Higher drug loading (e.g., p>5) may result in aggregation, insolubility, toxicity, or loss of cell permeability for some ADCs.

[0452] Typically, the amount of NMT inhibitor coupled to the antibody during the coupling reaction is less than the theoretical maximum. The antibody may contain, for example, many lysine residues that do not react with the NMT inhibitor-linker intermediate or linker reagent. Only the most reactive lysine groups can react with amine-reactive linker reagents. In addition, only the most reactive cysteine ​​thiol groups (e.g., sulfhydryl groups) can react with thiol-reactive linker reagents. In general, antibodies do not contain many (if any) free and reactive cysteine ​​thiol groups that may be connected to the NMT inhibitor. Most of the cysteine ​​thiol residues in the compound antibody exist in the form of disulfide bonds and must be reduced with a reducing agent (such as dithiothreitol (DTT) or TCEP) under partial or complete reducing conditions. The load of the ADC can be controlled in several different ways, including: (i) limiting the molar excess of the NMT inhibitor-linker intermediate or linker reagent relative to the antibody, (ii) limiting the coupling reaction time or temperature, and (iii) partial or limited reducing conditions for cysteine ​​thiol modification.

[0453] Some antibodies have reducible interchain disulfide bonds, i.e. cysteine ​​bridges. Antibodies can be made reactive to be coupled to linker reagents by treating with reducing agents such as DTT (dithiothreitol). In theory, each cysteine ​​bridge will therefore form two reactive thiol nucleophiles. Additional nucleophilic groups can be introduced into antibodies by reacting lysine with 2-iminothiolane (Traut reagent), resulting in amine conversion to thiol. Reactive thiol groups can be introduced into antibodies by engineering one, two, three, four or more cysteine ​​residues (e.g., preparing mutant antibodies comprising one or more non-natural cysteine ​​amino acid residues). US 7521541 teaches engineering antibodies by introducing reactive cysteine ​​amino acids. Cysteine ​​amino acids can be engineered at reactive sites of antibodies that do not form intrachain or intermolecular disulfide bonds (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 or drug linker reagent having a thiol-reactive electrophilic group of the present invention (e.g., maleimide or α-haloamide) to form an ADC having a cysteine ​​engineered antibody and a NMT inhibitor portion. The position of the NMT inhibitor portion can thus be designed, controlled, and understood. Since the engineered cysteine ​​thiol group typically reacts with a thiol-reactive linker reagent or drug linker reagent in high yield, the NMT inhibitor loading can be controlled. The IgG antibody was engineered to introduce a cysteine ​​amino acid by substitution at a single site in the heavy or light chain, resulting in two new cysteines on the symmetric antibody. NMT inhibitor loadings close to 2 and near-homogeneous conjugated product ADCs can be achieved.

[0454] Alternatively, site-specific coupling can be achieved by engineering the antibody to contain unnatural amino acids in its heavy and / or light chains, as described by Axup et al. ((2012), Proc Natl Acad Sci USA. 109(40): 16101-16116). Unnatural amino acids offer the additional advantage that orthogonal chemical reactions can be designed to connect linker reagents and NMT inhibitors.

[0455] When more than one nucleophilic or electrophilic group of an antibody reacts with an NMT inhibitor-linker intermediate or linker reagent and subsequently an NMT inhibitor, the resulting product is a mixture of ADC compounds, wherein the distribution of NMT inhibitors attached to the antibody is 1, 2, 3, etc. Liquid chromatography methods such as polymeric reverse phase (PLRP) and hydrophobic interaction (HIC) can separate the individual compounds in the mixture by NMT inhibitor loading value. ADC preparations with a single NMT inhibitor loading value (p) can be isolated, however, these single loading value ADCs may still be heterogeneous mixtures because the NMT inhibitor can be attached to different sites of the antibody via the linker.

[0456] Thus, the ADCs of the invention include mixtures of ADC compounds wherein the antibody bears one or more NMT inhibitor moieties, and wherein the NMT inhibitor moieties may be attached to the antibody at various amino acid residues.

[0457] In one embodiment, the drug loading (p) of the NMT inhibitor / antibody is between 1 and 10 NMT inhibitor / antibody. Suitably, the drug loading (p) is between 2 and 6, 4 and 6, 8 and 10, or 6 and 8 NMT inhibitor / antibody.

[0458] In one embodiment, the ADC of the present invention comprises the formula:

[0459]

[0460] Wherein Ab is an antibody as defined herein and p is as defined herein. Suitably, the ADC of the present invention or a salt thereof binds to the antibody via a sulfhydryl group on a cysteine ​​amino acid side chain on the antibody. Suitably, the antibody is trastuzumab or rituximab, in particular trastuzumab. Alternatively, the antibody is saccharotuzumab. Alternatively, the antibody is ifenatumomab. Suitably, p is between 1 and 10, for example, p is between 2 and 6, between 4 and 6, between 8 and 10, or between 6 and 8. Most suitably, p is, for example, about 5.

[0461] In one embodiment, the ADC of the present invention comprises the formula:

[0462]

[0463] Wherein Ab is an antibody as defined herein and p is as defined herein. Suitably, the ADC of the present invention or a salt thereof binds to the antibody via a sulfhydryl group on a cysteine ​​amino acid side chain on the antibody. Suitably, the antibody is trastuzumab or rituximab, in particular trastuzumab. Alternatively, the antibody is tactuzumab. Alternatively, the antibody is ifenatumomab. Suitably, p is between 1 and 10, for example, p is between 2 and 6, between 4 and 6, between 8 and 10, or between 6 and 8. Most suitably, p is, for example, about 5.

[0464] In one embodiment, the ADC of the present invention comprises the formula:

[0465]

[0466] wherein Ab is an antibody as defined herein and p is as defined herein. Suitably, the ADC of the invention or a salt thereof binds to the antibody via a sulfhydryl group on a cysteine ​​amino acid side chain on the antibody. Suitably, the antibody is trastuzumab or rituximab, in particular trastuzumab. Suitably, p is between 1 and 10, for example, p is between 2 and 6, between 4 and 6, between 8 and 10, or between 6 and 8. Most suitably, p is, for example, about 5.

[0467] In one embodiment, the ADC of the present invention comprises the formula:

[0468]

[0469] Wherein Ab is an antibody as defined herein and p is as defined herein. Suitably, the ADC of the present invention or a salt thereof binds to the antibody via a sulfhydryl group on a cysteine ​​amino acid side chain on the antibody. Suitably, the antibody is trastuzumab or rituximab, in particular trastuzumab. Alternatively, the antibody is saccharotuzumab. Alternatively, the antibody is ifenatumomab. Suitably, p is between 1 and 10, for example, p is between 2 and 6, between 4 and 6, between 8 and 10, or between 6 and 8. Most suitably, p is, for example, about 5.

[0470] In one embodiment, the ADC of the present invention comprises the formula:

[0471]

[0472] Wherein Ab is an antibody as defined herein and p is as defined herein. Suitably, the ADC of the present invention or a salt thereof binds to the antibody via a sulfhydryl group on a cysteine ​​amino acid side chain on the antibody. Suitably, the antibody is trastuzumab or rituximab, in particular trastuzumab. Alternatively, the antibody is saccharotuzumab. Alternatively, the antibody is ifenatumomab. Suitably, p is between 1 and 10, for example, p is between 2 and 6, between 4 and 6, between 8 and 10, or between 6 and 8. Most suitably, p is, for example, about 5.

[0473] In one embodiment, the ADC of the present invention comprises the formula:

[0474]

[0475] Wherein Ab is an antibody as defined herein and p is as defined herein. NMT is an NMT inhibitor. Suitably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Suitably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof. Suitably, the ADC of the present invention or a salt thereof binds to the antibody via a sulfhydryl group on the side chain of a cysteine ​​amino acid on the antibody. Suitably, the antibody is trastuzumab or rituximab, in particular trastuzumab. Alternatively, the antibody is saccharotuzumab. Alternatively, the antibody is ifenatumomab. Suitably, p is between 1 and 10, for example, p is between 2 and 6, between 4 and 6, between 8 and 10, or between 6 and 8. Most suitably, p is, for example, about 5.

[0476] Drug conjugates

[0477] The ADC of the present invention can be prepared using a drug conjugate or a salt and / or solvate thereof which is subsequently covalently bound to an antibody. Thus, in one embodiment, a drug conjugate or a salt and / or solvate thereof is provided which comprises a group capable of forming a covalent bond with a functional group such as a sulfhydryl group on the chain end (e.g., N-terminus) or amino acid side chain of an antibody.

[0478] In one embodiment, the drug conjugate has the formula:

[0479] A a -W w -Y y -NMT(Formula (X))

[0480] or a salt and / or solvate thereof,

[0481] in:

[0482] A is a first stretching unit, which, when present, comprises a group capable of forming a covalent bond with a functional group, such as a thiol group, on the chain end (such as the N-terminus) or the amino acid side chain of the antibody;

[0483] NMT is an NMT inhibitor as defined above; and

[0484] a, W, w, Y and y are as defined above.

[0485] It should be understood that the preferences and embodiments disclosed herein with respect to the ADC of formula (VII) are equally applicable to the drug conjugate of formula (X).

[0486] The NMT inhibitor forms a covalent bond with the second stretcher unit (Y) (when present), or when Y is absent, with an amino acid unit or a glucuronide unit (W). Suitably, the functional group on the NMT inhibitor is an amino group or an alcohol, such as an amino group. When the NMT inhibitor forms a covalent bond via an amino group, the amino group must have an available hydrogen atom to react with a corresponding functional group (e.g., a carbonyl group) in the linker, specifically in the second stretcher unit (Y), or when Y is absent, with an amino acid unit or a glucuronide unit (W), i.e., the amino group cannot be a tertiary amino group.

[0487] In one embodiment, the group capable of forming a covalent bond with a functional group on the chain terminus (eg, N-terminus) or amino acid side chain of an antibody is a maleimide moiety.

[0488] In one embodiment, A is:

[0489]

[0490] in Indicates the connection point with W.

[0491] In one embodiment, the drug conjugate is a compound of formula (DC-1):

[0492]

[0493] or its salts and / or solvates, wherein is an NMT inhibitor. Suitably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Suitably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof.

[0494] It should be understood by those skilled in the art that the phrase " "NMT inhibitor" refers to the portion remaining after an NMT inhibitor (e.g., an NMT inhibitor comprising a suitable functional group for attachment to a linker (e.g., an amino group (which comprises a hydrogen atom) or an alcohol (-OH))) reacts with a suitable functional group (e.g., a carbonyl group) on a linker to form a linker-NMT inhibitor covalent bond.

[0495] When the NMT inhibitor is a compound of formula (VI), any of the embodiments disclosed herein express:

[0496]

[0497] Where all variables are defined as above;

[0498] One of R5 and R6 is absent, so that the N atom can form a covalent bond with the linker.

[0499] Similarly, when the NMT inhibitor is a compound of formula (VIA), any of the embodiments disclosed herein express:

[0500]

[0501] Where all variables are defined as above;

[0502] One of R5 and R6 is absent, so that the N atom can form a covalent bond with the linker.

[0503] In one embodiment, the drug conjugate has the formula (DC-2):

[0504]

[0505] or its salts and / or solvates, wherein is an NMT inhibitor. Suitably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Suitably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof.

[0506] In one embodiment, the drug conjugate is a compound of formula (DC-5):

[0507]

[0508] or its salts and / or solvates, wherein is an NMT inhibitor. Suitably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Suitably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof.

[0509] In one embodiment, the drug conjugate is a compound of formula (DC-6):

[0510]

[0511] or its salts and / or solvates, wherein is an NMT inhibitor. Suitably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Suitably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof.

[0512] 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-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-[3-(3-{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy]ethoxy}propionylamino)propionylamino]phenoxy)-3,4-dihydroxy-2-methylcyclohexane-1-carboxylic acid:

[0513]

[0514] or a salt and / or solvate thereof.

[0515] In one embodiment, the drug conjugate is 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-methylcarbamic acid {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamino]-3-methylbutanoylamino]pentanoylamino]phenyl}methyl ester:

[0516]

[0517] or a salt and / or solvate thereof.

[0518] 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}propionylamino)propionylamino]phenoxy)-3,4-dihydroxy-2-methylcyclohexane-1-carboxylic acid:

[0519]

[0520] or a salt and / or solvate thereof.

[0521] In one embodiment, the drug conjugate is N-[({[(1-{[({[6-(3,4-difluoro-2-{2-[3-(1-hydroxy-

[0522] [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:

[0523]

[0524] or a salt and / or solvate thereof.

[0525] The drug conjugate may be provided in the form of a salt. The drug conjugate is provided in the form of a solvate. The drug conjugate may be provided in the form of a solvate of a salt. Suitably, a drug conjugate is provided.

[0526] The present invention also provides intermediate compounds that can be used to prepare drug conjugates.

[0527] In one embodiment, a compound of formula (ADC-I) is provided:

[0528]

[0529] Or a salt and / or solvate thereof, wherein NMT is an NMT inhibitor. Suitably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Suitably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof. Suitably, 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.

[0530] In one embodiment, a compound of formula (ADC-II) is provided:

[0531]

[0532] Or a salt and / or solvate thereof, wherein NMT is an NMT inhibitor. Suitably, the NMT inhibitor is a compound of formula (VI) or a salt thereof. Suitably, the NMT inhibitor is a compound of formula (VIA) or a salt thereof. Suitably, 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.

[0533] In one embodiment, the present invention provides a compound selected from the group consisting of:

[0534] {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamino]-3-methylbutanoylamino]pentanoylamino]phenyl}methyl 4-nitrophenyl carbonate;

[0535] (1S,2R,3S,4R,5R)-5-[2-(3-{[(9H-fluoren-9-yloxy)carbonyl]amino}propionamide)-4-({[(4-nitrophenoxy)carbonyl]oxy}methyl)phenoxy]-2,3,4-trihydroxycyclohexane-1-carboxylic acid;

[0536] (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}propionylamino)phenoxy)-2,3,4-trihydroxycyclohexane-1-carboxylic acid; and

[0537] (1S,2R,3S,4R,5R)-5-[2-(3-aminopropionylamino)-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,

[0538] or a salt and / or solvate thereof.

[0539] It should be understood that, in order to be used in medical treatment, the salt of the ADC of the present invention should be pharmaceutically acceptable. Non-pharmaceutically acceptable salts of the ADC of the present invention can be used in other situations, such as during the preparation of the ADC of the present invention. Suitable pharmaceutically acceptable salts will be clearly understood by those skilled in the art. Pharmaceutically acceptable salts include those described by 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 (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid) and organic acids (e.g., succinic acid, maleic acid, acetic acid, fumaric acid, citric acid, tartaric acid, benzoic acid, p-toluenesulfonic acid, methanesulfonic acid or naphthalenesulfonic acid). For example, other salts, such as oxalates or formates, can be used in the separation of the ADC of the present invention, and are included within the scope of the present invention.

[0540] Certain ADCs of the present invention can form acid addition salts with one or more equivalents of an acid. All possible stoichiometric and non-stoichiometric forms are included within the scope of the present invention.

[0541] It should be understood that the present invention encompasses all isomers of the ADC of the present invention, including all geometric, tautomeric and optical forms, and mixtures thereof (e.g., racemic mixtures). When additional chiral centers are present in the ADC of the present invention, all possible diastereomers, including mixtures thereof, are included within the scope of the present invention. Different isomeric forms can be separated or resolved from each other by conventional methods, or any given isomer can be obtained by conventional synthetic methods or by stereospecific or asymmetric synthesis.

[0542] The present disclosure includes all isotopic forms of the ADCs of the present invention provided herein, whether in the form of (i) 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) one or more atoms are substituted with atoms having the same atomic number as the predominant atoms in nature but a mass number different from the mass number of the latter (referred to herein as a "non-natural variant isotopic form"). It is understood that atoms may naturally exist in the form of a mixture of different mass numbers. The term "non-natural variant isotopic form" also includes embodiments in which the proportion of atoms of a given atomic number having a mass number that is less common in nature (referred to herein as "uncommon isotopes") is increased relative to the naturally occurring isotope, for example, to a level of >20%, >50%, >75%, >90%, >95% or >99% based on the number of atoms of that atomic number (the latter embodiment is referred to as an "isotopically enriched variant form"). The term "non-natural variant isotopic form" also includes embodiments in which the ratio of the uncommon isotope is reduced relative to the ratio of the naturally occurring isotope. Isotopic forms may include radioactive forms (i.e., they contain radioactive isotopes) and non-radioactive forms. Radioactive forms are typically isotopically enriched variant forms.

[0543] Therefore, the non-natural variant isotopic forms of the ADC of the present invention may contain one or more artificial or rare isotopes in one or more atoms, such as 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( 125 I), or may contain said isotope in one or more atoms in an increased proportion compared to the proportion predominant in nature.

[0544] For example, non-natural variant isotopic forms containing radioactive isotopes are useful in drug and / or substrate tissue distribution studies. 3H) and carbon-14 (i.e. 14 C) is particularly suitable for this purpose because it is easy to incorporate and can be easily detected. 2 Non-natural variant isotopic forms of H or D may offer certain therapeutic advantages due to greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, which may be preferred in some circumstances. In addition, non-natural variant isotopic forms can be prepared that contain positron emitting isotopes, such as 11 C. 18 F. 15 O and 13 N, which can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0545] In one embodiment, the ADC or salt thereof of the present invention is provided in a natural isotopic form.

[0546] In one embodiment, the ADC of the present invention or its salt is provided in a non-natural variant isotopic form. In a specific embodiment, the non-natural variant isotopic form refers to the incorporation of deuterium (i.e., 2 In one embodiment, the ADC of the present invention or its salt is in a non-radioactive isotopic form. In one embodiment, one or more atoms of the ADC of the present invention or its salt are in a radioactive isotopic form. Suitably, the radioisotope is a stable isotope. Suitably, the non-natural variant isotopic form is a pharmaceutically acceptable form.

[0547] In one embodiment, an ADC or a salt thereof of the present invention is provided, wherein a single atom of the compound exists in a non-natural variant isotopic form. In another embodiment, an ADC or a salt thereof of the present invention is provided, wherein two or more atoms exist in a non-natural variant isotopic form.

[0548] Non-natural isotopic variant forms can generally be prepared by conventional techniques known to those skilled in the art or by methods described herein (e.g., methods similar to the methods described in the accompanying Examples for preparing natural isotopic forms). Therefore, non-natural isotopic variant forms can be prepared by using suitable isotopic variant (or labeling) reagents instead of the normal reagents used in the Examples. Since the ADCs of the present invention are intended for use in pharmaceutical compositions, it is readily understood that each of them is preferably provided in a substantially pure form, such as at least 60% pure, more suitably at least 75% pure, preferably at least 85% pure, and especially at least 98% pure (% is based on weight). Impure compound preparations can be used to prepare purer forms used in pharmaceutical compositions.

[0549] In general, the ADC of the present invention can be prepared according to organic synthesis techniques known to those skilled in the art, as well as the representative methods listed below, the methods in the Examples, and their modified forms.

[0550] Uses of the ADC of the present invention

[0551] As used herein, the terms "prevent" or "prevention" mean to provide in advance and thus may involve preventing symptoms of a disease or disorder in a subject or preventing the recurrence of symptoms of a disease or disorder in a afflicted subject, and are not limited to preventing the disorder entirely.

[0552] As used herein, the term "treating" includes controlling, alleviating, reducing or regulating the disease state or symptoms thereof.

[0553] Inhibition of human NMT

[0554] As described above, inhibiting human NMT has been considered to be a goal for treating or preventing various diseases or conditions. The present invention provides ADCs comprising human NMT inhibitors. The term "human NMT inhibitor" used herein is intended to cover any portion that binds to human NMT. ​​Suitably, human NMT is HsNMT1. The inhibitor may act as a competitive inhibitor, or a partial competitive inhibitor. The inhibitor may bind to human NMT in the myr-CoA binding pocket or the peptide binding pocket (or inhibit human NMT by another mechanism). Since the ADC of the present invention or a pharmaceutically acceptable salt thereof contains an NMT inhibitor that is a human NMT inhibitor, it is expected that after the NMT inhibitor is released intracellularly from the ADC of the present invention, the NMT inhibitor will suitably bind to and inhibit human NMT through the peptide binding pocket.

[0555] Therefore, the ADC of the present invention or a pharmaceutically acceptable salt thereof is expected to be used for treating or preventing diseases or conditions associated with human NMT activity, or for treating or preventing diseases or conditions produced by targeting human NMT activity, such as hyperproliferative diseases such as cancer. Therefore, the present invention provides the ADC of the present invention or a pharmaceutically acceptable salt thereof for use as a medicament.

[0556] Also provided is an ADC of the invention, or a pharmaceutically acceptable salt thereof, for use in treating or preventing a disease or condition in which inhibition of human NMT provides a therapeutic or prophylactic effect. In one embodiment, an ADC of the invention, or a pharmaceutically acceptable salt thereof, is provided for use in treating a disease or condition in which inhibition of human NMT provides a therapeutic effect. In one embodiment, an ADC of the invention, or a pharmaceutically acceptable salt thereof, is provided for use in preventing a disease or condition in which inhibition of human NMT provides a prophylactic effect.

[0557] The present invention also provides a method for treating or preventing a disease or condition in a subject, wherein inhibiting human NMT can provide a therapeutic or preventive effect in a subject (e.g., a mammal, such as a human), the method 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 a method for treating a disease or condition in a subject, wherein inhibiting human NMT can provide a therapeutic effect in a subject (e.g., a mammal, such as a human), the method 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 a method for preventing a disease or condition in a subject, wherein inhibiting human NMT can provide a preventive effect in a subject (e.g., a mammal, such as a human), the method 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.

[0558] The present invention also provides the use of an ADC of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating or preventing a disease or condition in which inhibition of human NMT provides a therapeutic or preventive effect. The present invention also provides the use of an ADC of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating a disease or condition in which inhibition of human NMT provides a therapeutic effect. The present invention also provides the use of an ADC of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing a disease or condition in which inhibition of human NMT provides a preventive effect.

[0559] Hyperproliferative disorders

[0560] Since the ADC of the present invention has cytotoxic activity, it is expected that the ADC of the present invention or a pharmaceutically acceptable salt thereof can be used to treat or prevent hyperproliferative disorders.

[0561] Therefore, the present invention provides an ADC of the present invention or a pharmaceutically acceptable salt thereof for use in treating or preventing a hyperproliferative disorder. In a particularly suitable embodiment, the ADC of the present invention or a pharmaceutically acceptable salt thereof is used to treat a hyperproliferative disorder.

[0562] In one embodiment, the present invention provides the use of the ADC of the present invention or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating or preventing a hyperproliferative disorder. In a particularly suitable embodiment, the present invention provides the use of the ADC of the present invention or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a hyperproliferative disorder.

[0563] In one embodiment, the present invention provides a method for treating or preventing a hyperproliferative disorder in a subject, the method comprising administering a therapeutically effective amount of an ADC of the present invention or a pharmaceutically acceptable salt thereof. In a particularly suitable embodiment, the present invention provides a method for treating a hyperproliferative disorder in a subject, the method comprising administering a therapeutically effective amount of an ADC of the present invention or a pharmaceutically acceptable salt thereof.

[0564] In one embodiment, the hyperproliferative disorder is cancer.

[0565] In one embodiment, the cancer is a hematological malignancy selected from the group consisting of lymphoma (e.g., B-cell lymphoma, in particular 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 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 lentiginous melanoma).

[0566] Cancer can be additionally or alternatively 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 cancer (e.g., colon cancer), gallbladder cancer, kidney cancer, and liver cancer. For example, cancer can be ovarian serous cystadenocarcinoma, esophageal cancer, lung squamous cell carcinoma, lung adenocarcinoma, bladder urothelial carcinoma, uterine carcinosarcoma, gastric adenocarcinoma, breast invasive cancer, or hepatocellular carcinoma. In a suitable embodiment, cancer is breast cancer, such as triple-negative breast cancer or breast invasive cancer. In a suitable embodiment, cancer is brain cancer, breast cancer, prostate cancer, colon cancer, gallbladder cancer, or kidney cancer. In certain embodiments, cancer is breast cancer, colon cancer, or gallbladder cancer.

[0567] The cancer may additionally or alternatively be a blastoma, in particular a neuroblastoma, such as retinoblastoma, glioblastoma, small cell lung cancer or an astrocytoma.

[0568] In particularly suitable embodiments, the cancer may be selected from the group consisting of: a hematological malignancy (e.g., a lymphoma, in particular 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)), a solid tumor (e.g., brain, lung, breast (e.g., triple-negative breast cancer or invasive breast carcinoma), prostate, ovary, colorectal (e.g., colon), gallbladder, kidney, or liver cancer, or a neuroblastoma (e.g., retinoblastoma, glioblastoma, small cell lung cancer, or astrocytoma)), and a melanoma (e.g., superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, amelanotic melanoma, and acral lentiginous melanoma).

[0569] In suitable embodiments, cancer can be selected from the group consisting of diffuse large B cell lymphoma, Burkitt's lymphoma, multiple myeloma, neuroblastoma, AML and B acute lymphoblastic leukemia. In suitable embodiments, cancer can be selected from the group consisting of diffuse large B cell lymphoma, Burkitt's lymphoma, neuroblastoma, AML, B acute lymphoblastic leukemia and breast cancer. In suitable embodiments, cancer can be selected from the group consisting of diffuse large B cell lymphoma, neuroblastoma, B acute lymphoblastic leukemia and triple negative breast cancer. In suitable embodiments, cancer can be selected from the group consisting of diffuse large B cell lymphoma, Burkitt's lymphoma, multiple myeloma, neuroblastoma, AML, B acute lymphoblastic leukemia and triple negative breast cancer. In suitable embodiments, cancer can be selected from the group consisting of diffuse large B cell lymphoma, Burkitt's lymphoma, multiple myeloma, neuroblastoma, AML, B acute lymphoblastic leukemia and triple negative breast cancer. In suitable embodiments, cancer can be selected from the group consisting of multiple myeloma, neuroblastoma, AML, B acute lymphoblastic leukemia and triple negative breast cancer. In suitable embodiments, the cancer may be selected from the group consisting of multiple myeloma, neuroblastoma, and triple negative breast cancer.

[0570] In a suitable embodiment, the cancer expresses HER2 protein, i.e., the cancer contains cells with HER2 protein on the cell surface. HER2 protein is overexpressed in a variety of tumors and can be assessed using methods commonly used in the art, such as using immunohistochemistry staining (IHC) to assess overexpression of HER2 protein, or using fluorescent in situ hybridization (FISH) to assess the amplification of the HER2 gene.

[0571] Suitably, the cancer expressing the 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 tumors, 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, renal cancer, vulvar cancer, thyroid cancer and penile cancer, e.g. breast cancer.

[0572] In one embodiment, the cancer is HER2-positive breast cancer.

[0573] In a suitable embodiment, cancer expresses CD20. The CD20 expression of cancer cells can be assessed using methods commonly used in the art, such as immunohistochemical staining (IHC). Suitably, the cancer expressing CD20 is a lymphoma (e.g., high-grade mantle zone lymphoma, follicular lymphoma, plasmablastic lymphoma, diffuse large B-cell lymphoma, or Burkitt's lymphoma), a myeloma (e.g., multiple myeloma), or a leukemia (e.g., chronic lymphocytic leukemia, acute myeloid leukemia (AML), or B acute lymphocytic leukemia).

[0574] In a suitable embodiment, the cancer is a MYC-addicted cancer as described in WO2020 / 128475, the entire contents of which are incorporated herein by reference for the purpose of defining MYC-addicted cancer.

[0575] Combination therapy

[0576] Although the ADC of the present invention or its pharmaceutically acceptable salt can be used as the only active ingredient in a drug, the ADC of the present invention or its pharmaceutically acceptable salt can also be used in combination with one or more other therapeutic agents. Therefore, the present invention also provides the ADC of the present invention or its pharmaceutically acceptable salt in combination with another therapeutic agent. The other therapeutic component can be administered simultaneously, sequentially or separately. The present invention also provides a kit consisting of multiple parts, the multiple parts comprising: (a) a first pharmaceutical composition comprising the ADC of the present invention or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier; and (b) a second pharmaceutical composition comprising another therapeutic agent and a pharmaceutically acceptable carrier. Such additional therapeutic agents can be the ADC of the present invention or its pharmaceutically acceptable salt.

[0577] The ADC of the present invention or a pharmaceutically acceptable salt thereof can be used in combination with one or more additional therapeutic agents that can be used to treat or prevent hyperproliferative disorders, such as cancer or another disease or disorder in which inhibition of human NMT provides a therapeutic or preventive effect. The components of such a combination can be administered separately at different times during the course of treatment or simultaneously in separate or single combinations. The present invention should therefore be understood to include all such regimens of simultaneous or alternating treatment, and the term "administering" should be interpreted accordingly. It should be understood that the scope of the combination of the ADC of the present invention or a pharmaceutically acceptable salt thereof with other therapeutic agents for the treatment or prevention of a disease or disorder (in which inhibition of human NMT provides a therapeutic or preventive effect) in principle includes any combination with any pharmaceutical composition for the treatment or prevention of a disease or disorder (in which inhibition of human NMT provides a therapeutic or preventive effect).

[0578] When used in combination with the ADC of the present invention or a pharmaceutically acceptable salt thereof, the additional therapeutic agent may be used in an amount indicated for the agent in, for example, the Physicians' Desk Reference (PDR), or in an amount otherwise determined by a person of ordinary skill in the art. When the ADC of the present invention or a pharmaceutically acceptable salt thereof is used in combination with one or more additional therapeutic agents, whether used simultaneously or sequentially, the following combination ratios and dosage ranges are suitable: When used in combination with another therapeutic agent, the weight ratio of the ADC of the present invention or a pharmaceutically acceptable salt thereof to the other therapeutic agent used may be, for example, in the range of about 10:1 to about 1:10.

[0579] In one embodiment, when the ADC of the present invention or a pharmaceutically acceptable salt thereof is used to treat or prevent cancer, the ADC of the present invention or a pharmaceutically acceptable salt thereof can be used in combination with one or more additional therapeutic agents simultaneously or sequentially for treating or preventing cancer. More suitably, when the ADC of the present invention or a pharmaceutically acceptable salt thereof is used to treat cancer, the ADC of the present invention or a pharmaceutically acceptable salt thereof can be used in combination with one or more additional therapeutic agents simultaneously or sequentially for treating cancer.

[0580] 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.

[0581] Suitably, the chemotherapeutic agent is selected from the group consisting of fluorouracil (5-FU), pertuzumab, paclitaxel, carboplatin, cisplatin, gemcitabine, capecitabine, irinotecan (CPT-11), paclitaxel, docetaxel, pemetrexed, sorafenib, vinblastine, vinorelbine, everolimus, tanespimycin, bevacizumab, oxaliplatin, lapatinib, ado-trastuzumab emtansine (TDM1) or a drug described in International Publication No. WO2003 / 038043, an LH-RH analog (e.g. leuprolide or goserelin), estramustine phosphate, an estrogen antagonist (e.g. tamoxifen or raloxifene) and an aromatase inhibitor (e.g. anastrozole, letrozole or exemestane).

[0582] Such combined therapy can be achieved by administering the ADC of the present invention or its pharmaceutically acceptable salt and one or more other therapeutic agents simultaneously, sequentially or separately. Such combination products can be used in any suitable dosage range, such as the ADC of the present invention or its pharmaceutically acceptable salt in the dosage range described herein, and other pharmaceutically active agents can be in their approved dosage range.

[0583] Dosage and preparation

[0584] It will be appreciated by those skilled in the art that the appropriate dose of the ADC of the present invention or its pharmaceutically acceptable salt and the appropriate dose of the composition comprising the ADC of the present invention or its pharmaceutically acceptable salt may vary from patient to patient. Determining the optimal dose will generally involve balancing the level of therapeutic benefit and any risk or harmful side effects. The selected dose level will depend on a variety of factors, including but not limited to the activity of the specific ADC, the route of administration, the time of administration, the excretion rate of the ADC, the duration of treatment, other drugs, ADCs and / or materials used in combination, the severity of the condition, and the species, sex, age, weight, condition, general health and previous medical history of the patient. The amount and route of administration of the ADC will ultimately be determined by a physician, veterinarian or clinician, but the selected dose will generally obtain a local concentration at the site of action, achieving the desired effect without causing substantially harmful or toxic side effects.

[0585] Administration can be achieved in a single dose, continuously or intermittently (e.g., at appropriate time intervals, in divided doses) throughout the course of treatment. Methods for determining the most effective mode of administration and dosage 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 performed according to the dosage level and pattern selected by the treating physician, veterinarian, or clinician.

[0586] Typically, a suitable dose of ADC is in the range of about 100 ng to about 25 mg (more typically about 1 μg to about 10 mg) per kilogram of subject body weight per day. When the ADC is a salt, ester, amide, prodrug, etc., the dosage is calculated based on the parent ADC, so the actual weight to be used is increased proportionally.

[0587] In one embodiment, the ADC of the present invention or a pharmaceutically acceptable salt thereof is administered to a human patient at a dosage regimen of about 100 mg, 3 times a day. In one embodiment, the ADC of the present invention or a pharmaceutically acceptable salt thereof is administered to a human patient at a dosage regimen of about 150 mg, 2 times a day. In one embodiment, the ADC of the present invention or a pharmaceutically acceptable salt thereof is administered to a human patient at a dosage regimen of about 200 mg, 2 times a day. In one embodiment, the ADC of the present invention or a pharmaceutically acceptable salt thereof is administered to a human patient at a dosage regimen of about 50 or about 75 mg, 3 or 4 times a day. In one embodiment, the ADC of the present invention or a pharmaceutically acceptable salt thereof is administered to a human patient at a dosage regimen of about 100 or about 125 mg, 2 times a day.

[0588] The above dosages may apply to the ADC of the present invention or a pharmaceutically acceptable salt thereof (including an NMT inhibitor and an antibody linker) or to an effective amount of the NMT inhibitor provided, such as the amount of the NMT inhibitor that can be released after cleavage of the linker.

[0589] For the prevention or treatment of disease, the appropriate dose of the ADC of the present invention will depend on the type of disease to be treated, the severity and course of the disease, whether the molecule is administered for the purpose of prevention or treatment, previous therapy, the patient's clinical history and response to antibodies, and the judgment of the attending physician. The ADC of the present invention or its pharmaceutically acceptable salt is suitable for administration to the patient once or through a series of treatments. Depending on the type and severity of the disease, about 1 μg / kg to 15 mg / kg (e.g., 0.1-20 mg / kg) of the ADC of the present invention or its pharmaceutically acceptable salt is the initial candidate dose for administration to the patient, for example, whether or not by one or more separate administrations or by continuous infusion. The typical daily dose range can be about 1 μg / kg to 100 mg / kg or more, depending on the factors mentioned above. An exemplary dose of the ADC of the present invention or its pharmaceutically acceptable salt administered to a patient is in the range of about 0.1 to about 10 mg / kg of the patient's body weight. For repeated administration over several days or longer (depending on the condition), the treatment is maintained until the desired inhibition of disease symptoms occurs. An exemplary dosing regimen includes administering an initial loading dose of about 4 mg / kg, followed by an additional dose of the ADC of the present invention or a pharmaceutically acceptable salt thereof, administered weekly, biweekly, or every three weeks. Other dosing regimens may be used. The progress of this treatment is easily monitored by conventional techniques and assays.

[0590] Although the active ingredient can be administered alone, it is preferably present in a pharmaceutical preparation or composition. Therefore, the present invention provides a pharmaceutical preparation or composition comprising the ADC of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent, excipient or carrier (collectively referred to herein as "carrier" material). The pharmaceutical composition of the present invention can be in the form of a pharmaceutical preparation.

[0591] Therefore, in one embodiment, the present invention provides a pharmaceutical composition comprising the ADC of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0592] In one embodiment, a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) is provided for treating or preventing a disease or condition as described herein. In one embodiment, a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) is provided for treating a disease or condition as described herein. In one embodiment, a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) is provided for preventing a disease or condition as described herein.

[0593] In a further embodiment, a method for treating or preventing a disease or condition as described herein is provided, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt). In a further embodiment, a method for treating a disease or condition as described herein is provided, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt). In a further embodiment, a method for preventing a disease or condition as described herein is provided, comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt). The pharmaceutical composition of the present invention may be in the form of a pharmaceutical preparation as described below.

[0594] The present invention also provides the use of a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) in the manufacture of a medicament for treating or preventing a disease or condition described herein. The present invention also provides the use of a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) in the manufacture of a medicament for treating a disease or condition described herein. The present invention also provides the use of a pharmaceutical composition comprising an ADC of the present invention or a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt) in the manufacture of a medicament for preventing a disease or condition described herein.

[0595] Suitably, the disease or disorder is a hyperproliferative disorder, such as cancer. In a suitable embodiment, the cancer is a cancer expressing the HER2 protein.

[0596] The ADC of the present invention or its pharmaceutically acceptable salt can be administered by intravenous, cutaneous, intramuscular or subcutaneous injection or infusion. The active ingredient will be present in the form of an aqueous solution acceptable to the parenteral, the solution being pyrogen-free and having a suitable pH value, isotonicity and stability. Those skilled in the art are fully capable of preparing suitable solutions using, for example, isotonic vehicles (e.g., sodium chloride injection, Ringer's injection, lactated Ringer's injection), preservatives, stabilizers, buffers, antioxidants and / or other additives as needed.

[0597] In one embodiment, the pharmaceutical composition further comprises other active agents, such as other therapeutic or prophylactic agents.

[0598] Suitable carriers, diluents, excipients, etc. can be found in standard pharmaceutical texts. See, for example, Handbook of Pharmaceutical Additives, 2nd edition (M. Ash and I. Ash, eds.), 2001 (Synapse Information Resources, Inc., Endicott, New York, USA), Remington's Pharmaceutical Sciences, 20th edition, Lippincott, Williams & Wilkins, 2000; and Handbook of Pharmaceutical Excipients, 2nd edition, 1994.

[0599] Another aspect of the present invention relates to a method for preparing a pharmaceutical composition, comprising mixing at least one [11C]-radiolabeled ADC of the present invention or a pharmaceutically acceptable salt thereof with one or more other pharmaceutically acceptable ingredients known to those skilled in the art (e.g., carriers, diluents, excipients, etc.). If formulated as discrete units (e.g., tablets, etc.), each unit contains a predetermined amount (dose) of the active compound.

[0600] The term "pharmaceutically acceptable" as used herein relates to compounds, ingredients, materials, compositions, dosage forms, etc. that are suitable for use in contact with the tissues of relevant subjects (e.g., humans) within the scope of sound medical judgment without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio. Each carrier, diluent, excipient, etc. must also be "acceptable" from the perspective of compatibility with the other ingredients of the formulation.

[0601] The preparation can be prepared by any method known in the pharmaceutical field. These methods include the step of combining the ADC of the present invention or its pharmaceutically acceptable salt with a carrier constituting one or more auxiliary components. Generally, the preparation is prepared by uniformly and closely combining the ADC of the present invention or its pharmaceutically acceptable salt with a carrier (e.g., a liquid carrier, a subdivided solid carrier, etc.), and then molding the product (if necessary).

[0602] Preparations 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 also contain other pharmaceutically acceptable ingredients, such as antioxidants, buffers, preservatives, stabilizers, antibacterial agents, suspending agents, thickeners, and solutes, so that the preparation is isotonic with the blood (or other relevant body fluids) of the intended recipient. Examples of excipients include, for example, water, alcohol, polyols, glycerol, vegetable oils, etc. Examples of suitable isotonic carriers for use in such preparations include sodium chloride injection, Ringer's solution, or lactated Ringer's injection. Typically, the concentration of the ADC of the present invention or its pharmaceutically acceptable salt in the liquid is from about 1 ng / ml to about 10 μg / ml, for example, 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 a sterile liquid carrier, such as water for injection, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.

[0603] The formulations may be prepared to provide fast or slow release; immediate, delayed, timed or sustained release; or a combination thereof.

[0604] In some embodiments, the ADC of the present invention or its pharmaceutically acceptable salt can be administered orally. The pharmaceutical composition for oral administration can be in tablet, capsule, powder or liquid form. The tablet can contain a solid carrier or adjuvant. The liquid pharmaceutical composition usually contains a liquid carrier, such as water, petroleum, animal or vegetable oil, mineral oil or synthetic oil. It can include physiological saline solution, glucose or other sugar solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. The capsule can contain a solid carrier, such as gelatin.

[0605] In a suitable embodiment, administration is by infusion or injection, particularly bolus injection.

[0606] The ADC of the present invention or a pharmaceutically acceptable salt thereof is expected to exhibit one or more of the following advantageous properties:

[0607] - in vivo cytotoxic activity, such as demonstrated in the mouse xenograft model of Biological Example 2; and

[0608] - in vivo tolerance, such as demonstrated in the mouse xenograft models of Biological Examples 2, 3, 10, 11, 12 and 13 and the monkey model of Biological Example 9;

[0609] - in vitro cytotoxicity against various cancer cell lines, such as shown in Biological Examples 4, 6, 7 and 14;

[0610] - Improved efficacy on gastric cancer organoids compared to known controls as shown in Biological Example 8; and

[0611] - As shown in Biological Example 6, the bystander effect is improved.

[0612] Such properties are expected to make the ADCs of the invention, or pharmaceutically acceptable salts thereof, useful in treating or preventing (eg, treating) hyperproliferative disorders, such as cancer or other diseases or disorders in which inhibition of human NMT provides a therapeutic or preventive effect.

[0613] Example

[0614] Abbreviations

[0615] ALT Alanine aminotransferase

[0616] AST Aspartate aminotransferase

[0617] ALP alkaline phosphatase

[0618] GGT γ-glutamyltransferase

[0619] CK Creatine kinase

[0620] LDH Lactate dehydrogenase

[0621] TP Total Protein

[0622] ALB albumin

[0623] GLO Globulin

[0624] A / G Albumin / Globulin Ratio

[0625] TBIL Total bilirubin

[0626] BU blood urea nitrogen

[0627] CRE Creatinine

[0628] BUN / C blood urea nitrogen / creatinine ratio

[0629] GLU Glucose

[0630] CHO Cholesterol

[0631] TG triglyceride

[0632] Na Sodium

[0633] K Potassium

[0634] Cl Chloride

[0635] Ca Calcium

[0636] P Phosphate

[0637] WBC

[0638] ABNEUT Neutrophils

[0639] ABLYMP lymphocytes

[0640] ABMONOmonocytes

[0641] ABBASO basophils

[0642] ABEOS Eosinophils

[0643] PLT Platelets

[0644] MPV Mean platelet volume

[0645] RBC Red blood cells

[0646] HCT Hematocrit

[0647] HGB Hemoglobin

[0648] MCV mean corpuscular volume

[0649] MCH mean corpuscular hemoglobin

[0650] MCHC mean corpuscular hemoglobin concentration

[0651] ABRETIC Reticulocyte

[0652] QW Once a week

[0653] Synthesis of Example Compounds

[0654] General experimental details

[0655] LC-MS

[0656] Compounds requiring purification under basic conditions were typically purified on an LC-MS system equipped with a YMC Actus Triart C18 5 μm (20 x 250 mm) column or a Gemini NX 5 μm C18 (100 x 30 mm) column using a gradient elution of acetonitrile in water containing 20 mM ammonium bicarbonate (10-45% in 30 min, then 95% acetonitrile for 2 min).

[0657] HPLC

[0658] The purity of some compounds was determined by TyeEclipse Extend or XDB 5μm C18 (150x 4.6mm), Xbridge 5μm C18 (100x 4.6mm), Zorbax Extend 5μm C18 (150x 4.6mm) or Shimadzu L column 2ODS 5μm C18 (150x4.6mm) column using a gradient elution of acetonitrile in water containing 10mM ammonium acetate for 15 minutes (HPLCB), 17 minutes (B1) and 18 minutes (B3).

[0659] The purity of some compounds was determined by analytical HPLC using a Poroshell 120 2.7 μm EC18 (100 x 4.6 mm), Luna Omega Polar 3 μm C18 (100 x 4.6 mm), Xbridge 5 μm C18 (150 x 4.6 mm) or Sunfire 5 μm C18 (100 x 4.6 mm) 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).

[0660] The purity of some compounds was determined by analytical HPLC using a Gemini NX 3 μm C18 (100 x 4.6 mm) column with a gradient of acetonitrile in water containing 0.05% formic acid over 16 minutes (A6).

[0661] NMR

[0662] 1 H NMR and 13 C spectra were recorded at room temperature on 400 MHz and 101 MHz instruments, respectively, and were referenced to the residual solvent signal unless otherwise stated. Data are presented as follows: chemical shift (ppm), integration, multiplicity (br = broad, s = singlet, d = doublet, t = triplet, q = quartet, p = quintet, m = multiplet) and coupling constant (Hz).

[0663] ADC Test Methods

[0664] SEC-HPLC

[0665] Column: TOSOH TSKgel G3000SWXL 7.8mm x 30cm 5μm particles (MERCK808541) combined with a safety guard column (MERCK 822858) equipped with a GFC3000 4x3mm column core (Phenomenex); Buffer: 0.2M phosphate, 0.25M KCl, 10% IPA; Gradient: isocratic at 25°C, 0.5ml / min. The sample load was approximately 10μg, and the monomer and concentration were determined by the 214nm signal. Monomers are reported based on peak integration, and [ADC] mg / mL is based on the antibody calibration curve.

[0666] RP-HPLC for residual NMT inhibitors

[0667] Chromatographic column: 2.6μm C8 LC column 50 x 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:

[0668] time ％B 0.00 5 8.00 95 8.10 100 9.00 100 9.10 5 10.00 5

[0669] 50μl sample (ADC or PBS / PS20 matrix) + 2μl 5M NaCl + 150μl cold MeOH (from -20C freezer). Incubate at -20°C for 30 minutes. Centrifuge at 21,000g for 30 minutes at 4°C. Extract 125μl supernatant and mix with 125μl WFI. Inject 100μl of it onto the Kinetex column. Analyze the data at 214nm, and estimate the residual NMT inhibitor in the sample based on the external calibration curve of the relevant NMT inhibitor-linker. The results are expressed as the percentage of free relative to free and bound, and the amount of bound NMT inhibitor is determined using the ADC concentration and the calculated DAR.

[0670] HIC-HPLC for mean DAR (drug-to-antibody ratio) calculation

[0671] This method can be used as an alternative to the PLRP-HPLC method to determine the average DAR.

[0672] Column: TOSOH Butyl-NPR 4.6mm x 3.5cm, 2.5μm particle size (Merck822855); Mobile phase A: 1.5M (NH4)2SO4, 25mM NaPi, pH 6.95±0.05; Mobile phase B: 25mM NaH2PO4 pH 6.95±0.05+25% IPA; Gradient: 25℃ 0.8ml / min:

[0673] time ％B 0 0 12 100 12.1 0 18 0

[0674] Load cartridge. 10 μg and report results / analyze at 214 nm.

[0675] RP-HPLC for average DAR calculation

[0676] Chromatographic column: PLRP-S 2.1 mm x 5 cm, 5 μm (Agilent PL1912-1502); Mobile phase A: 0.1% TFA in water; Mobile phase B: 0.1% TFA in acetonitrile; Gradient: 80°C, 1 mL / min:

[0677] time ％B 0 22.5 2 22.5 21.5 49.5 22.5 90.0 26.5 90.0 27.5 22.5 32.0 22.5

[0678] About 10ug sample (ADC) + 5μl 0.1M DTT was supplemented to 50μL with 0.5M Tris, pH 8.0 and incubated at 37°C for 15 minutes. The sample was then diluted 1:1 (+50μL) with 49% water, 49% acetonitrile, 2% formic acid. 20uL of this solution was then injected onto the RP-HPLC column. Data was analyzed at 214nm and the average DAR was calculated.

[0679] Endotoxin kinetic chromogenic assay

[0680] Endotoxin was determined by kinetic chromogenic LAL assay using the Endosafe PTS Endotoxin System. ADC was diluted 10-fold with LAL reagent water. All samples were analyzed on 0.01–1 EU / mL cartridges. EU / mL values ​​were divided by ADC[P] mg / mL to convert to EU / mg.

[0681] 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

[0682]

[0683] Step 1-Intermediate (2): 1,5-dimethyl-1H-pyrazole-3-carboxylic acid

[0684] 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) at room temperature was added ethanol (0.4 ml) and LiOH.H2O (9.985 g, 237.968 mmol). The resulting mixture was stirred at room temperature for 16 hours. TLC / LCMS showed that SM was completely consumed. The reaction mixture was acidified with 3N HCl solution (pH about 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 light yellow solid (16 g, 99%). 1 H NMR(400MHz, DMSO)d 12.42(s,1H),6.45(s,1H),3.77(s,3H),2.25(s,3H).

[0685] Step 2-Intermediate (3): N-methoxy-N,1,5-trimethyl-1H-pyrazole-3-carboxamide

[0686] 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 room temperature for 16 hours. TLC was checked and showed product formation. The reactant 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 product was purified by combiflash using 5% MeOH in DCM to give N-methoxy-N,1,5-trimethyl-1H-pyrazole-3-carboxamide (3) as a light yellow solid (15.0 g, 69.05%). 1 H NMR (400 MHz, DMSO) d 6.41 (s, 1H), 3.76 (s, 3H), 3.67 (s, 3H), 3.32 (s, 3H), 2.26 (s, 3H); LCMS (NH4Oac:CAN): M+H = 184 Rt = 2.17 min, run time 5 min.

[0687] Step 3-Intermediate (4): 1-(1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one

[0688] 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 tert-butyl lithium (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 was checked to show product formation and the reaction mixture was quenched with saturated NH4Cl solution. The 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 product was purified by combiflash chromatography using 5% MeOH in DCM to afford 1-(1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one (4) as a light yellow solid (6.0 g, 40.63%). 1 H NMR (400 MHz, DMSO) d 6.45 (s, 1H), 3.79 (s, 3H), 2.25 (s, 3H), 1.31 (s, 9H); LCMS (HCOOH: A N ): M+H = 181, Rt = 1.86 min, run time 3 min.

[0689] Step 4-Intermediate (5): 1-(4-bromo-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one

[0690] 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) in portions under ice-cold conditions. The resulting reaction mixture was stirred at RT for 16 hours. TLC and LCMS were checked to show product formation. 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 in vacuo 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. 1 H NMR (400 MHz, DMSO) d 3.86 (s, 3H), 2.25 (s, 3H), 1.30 (s, 9H); LCMS (NH4Oac:CAN): M+H = 259, Rt = 3.59 min, running time 5 min.

[0691] Step 5-Intermediate (6): 1-(1,5-dimethyl-4-vinyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one

[0692] 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 tributyl vinyl tin (17.2 ml, 54.257 mmol) at room temperature. The reaction mixture was then purged with argon 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 was checked, showing that the starting material had been consumed and the desired product was formed. The reaction mixture was then diluted with ethyl acetate and washed with potassium fluoride solution, the precipitate was filtered through a filter cartridge and washed with water and brine, dried over sodium sulfate and concentrated. The crude product 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%). 1 H NMR (400 MHz, DMSO) d 6.96-6.89 (m, 1H), 5.29-5.20 (m, 2H), 3.82 (s, 3H), 2.31 (s, 3H), 1.30 (s, 9H); LCMS (HCOOH: A N ): M+H = 207, Rt = 2.19 min, run time 3 min.

[0693] Step 6-Intermediate (7): 2-(1,5-dimethyl-3-pivaloyl-1H-pyrazol-4-yl)acetaldehyde

[0694] 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 used in the next step without purification. 1HNMR (400 MHz, DMSO) d 9.49 (s, 1H), 3.82 (s, 3H), 3.65 (s, 2H), 2.17 (s, 3H), 1.30 (s, 9H); LCMS (NH4Oac:CAN): M+H = 223, Rt = 1.86 min, running time 3 min.

[0695] Step 7-Intermediate (8): 1-(4-(2-hydroxyethyl)-1,5-dimethyl-1H-pyrazol-3-yl)-2,2-dimethylpropan-1-one

[0696] 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) in portions under ice-cold 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 and diluted with ethyl acetate, washed with water, brine and concentrated in vacuo to give the crude product. This batch was purified by combi flash 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. 1 H NMR (400 MHz, DMSO) d 4.46 (t, 1H), 3.77 (s, 3H) 3.36 (t, 2H), 2.69 (t, 2H), 2.17 (s, 3H), 1.30 (s, 9H); LCMS (HCOOH: CA N): M+H = 225, Rt = 1.81 min, run time 3 min.

[0697] 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

[0698] 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]pyridin-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 showed product formation 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 product 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 viscous gum (3.0 g, 49.05%). 1 H NMR (400 MHz, DMSO) d 8.50-8.42 (brs, 1H), 7.62 (s, 1H), 7.53 (d, 1H), 7.30-7.15 (m, 3H), 4.76 (s, 2H), 3.92 (t, 2H), 3.67 (s, 3H), 2.81 (t, 2H), 2.67 (s, 3H), 1.86 (s, 3H), 1.32 (s, 9H), 1.19 (s, 9H); LCMS (HCOOH: CA N): M+H=596, Rt=1.75 min, run time 5 min.

[0699] 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

[0700] 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]pyridin-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, 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%). 1 H NMR (400 MHz, DMSO)d 8.55-8.45 (brs, 1H), 7.63 (s, 1H), 7.60 (d, 1H), 7.38 (d, 1H), 7.35-7.25 (brs, 2H), 4.78 (s, 2H), 4.60 (d, 1H), 4.07 (d, 1H), 3.92-3.88 (m, 1H), 3.84-3.78 (m, 1H), 3.52 (s, 3H), 2.80-2.70 (m, 1H), 2.68 (s, 3H), 2.60-2.52 (brs, 1H), 1.78 (s, 3H), 1.34 (s, 9H), 0.74 (s, 9H); LCMS (NH4Oac:CAN): M+H=598, Rt=3.75 minutes, running for 5 minutes.

[0701] 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

[0702] 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 2M HCl in diethyl ether (40.0 ml) at 0°C. The reaction mixture was stirred at room temperature for 3 hours. TLC and LCMS showed that the starting material had been consumed. The reaction mixture was evaporated under reduced pressure to obtain the crude product. The crude product was triturated with 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 light yellow solid (HCl salt) (1.04 g, 96.93 mmol, 49%). 1 H NMR (400 MHz, DMSO) d 10.02-9.96 (brs, 2H), 9.26 (s, 1H), 8.44 (s, 1H) 8.08 (d, 1H), 7.98 (d, 1H), 7.64 (t, 1H), 7.46-7.40 (m, 1H), 4.75 (s, 3H), 4.16 (s, 1H), 3.97 (t, 2H), 3.71 (s, 3H), 2.87-2.80 (m, 1H), 2.72-2.65 (m, 1H), 2.60 (s, 3H), 2.09 (s, 3H), 0.76 (s, 9H); LCMS (HCOOH: CA N): M+H=498, Rt=2.54 min, run 5 min; HPLC RT (B3) 8.739 min.

[0703] The following compounds were prepared using methods analogous to those described for NMT inhibitor 1:

[0704] Table 1: NMT inhibitors

[0705]

[0706]

[0707]

[0708]

[0709]

[0710]

[0711]

[0712]

[0713] NMT inhibitors 1 to 25 can be prepared according to the methods disclosed in European Patent Application No. 22194959.7, the entire contents of which are incorporated herein by reference to describe the synthesis and activities of NMT inhibitors.

[0714] Synthesis of drug conjugate 1: 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-methylcarbamic acid {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamino]-3-methylbutanoylamino]pentanoylamino]phenyl}methyl ester:

[0715]

[0716] To {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido]-3-methylbutanamido]pentanamido]phenyl}methyl carbonate 4-nitrophenyl ester hydrate (ADC intermediate 1, such as Synlett, 2009, No. 18, pp. 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, 12 mg, 16 mmol) in anhydrous DMF (2 mL) using standard carbonate formation conditions as 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 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-methylcarbamic acid {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamino]-3-methylbutanoylamino]pentanoylamino]phenyl}methyl ester (drug conjugate 1), a white solid after lyophilization (10.5 mg). Drug conjugate 1 can be used to prepare ADC by reacting with an antibody using methods known to those skilled in the art and described herein (e.g., methods described for ADC Example 1 or ADC Example 2).

[0717] 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-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}--2-[3-(3-{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy]ethoxy}propionamido)propionamido]phenoxy)-3,4-dihydroxy-2-methylcyclohexane-1-carboxylic acid (hydrate)

[0718]

[0719] Step 1: To a solution of (1S,2R,3S,4R,5R)-5-[2-(3-{[(9H-fluoren-9-yloxy)carbonyl]amino}propionylamino)-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 reactant was stirred at room temperature (22°C). After 16 h, the mixture was purified directly 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}propionylamino)phenoxy)-2,3,4-trihydroxycyclohexane-1-carboxylic acid (ADC intermediate 3) as a white solid (107 mg) after lyophilization.

[0720] 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}propionylamino)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 solution, 0.5 mL) was added dropwise at room temperature. The mixture was stirred at room temperature for 8 hours. HCl (4N dihydrate) was added. The mixture was purified by RP-HPLC to give (1S,2R,3S,4R,5R)-5-[2-(3-aminopropionylamino)-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 after lyophilization (TFA salt, 42 mg).

[0721] Step 3: To a solution of (1S,2R,3S,4R,5R)-5-[2-(3-aminopropionylamino)-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 hour 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}propionylamino)phenoxy)-2,3,4-trihydroxycyclohexane-1-carboxylic acid hydrate (drug conjugate 2) as a white solid (36 mg) after lyophilization.

[0722] Synthesis of drug conjugate 3

[0723]

[0724] 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 hours, the mixture was directly purified by RP-HPLC to obtain ADC intermediate 5, which was a white solid after lyophilization.

[0725] Step 2: Dissolve ADC intermediate 5 in acetonitrile / water (6 / 4, v / v), and add NaOH (1N, aqueous solution) dropwise at room temperature. Stir the mixture at room temperature for 8 hours. Add HCl (4N dihydrate) The mixture was purified by RP-HPLC and lyophilized to give a white solid (ADC intermediate 6).

[0726] Step 3: Mal-PEG2-OSu was added to a solution of ADC intermediate 6 in acetonitrile / water (6 / 4, v / v), followed by DIEA. The reaction mixture was stirred at room temperature for 1 hour, directly purified by RP-HPLC, and lyophilized to obtain drug conjugate 3 as a white solid.

[0727] ADC Example 1 - Preparation of Trastuzumab-NMT Inhibitor ADC (DAR 5)

[0728] Trastuzumab was purchased and reconstituted into a 25 mg / mL solution. Prior to reduction and coupling, 5% v / v of 500 mM Tris, 25 mM EDTA, pH 8.5 was added to adjust the pH. 2.5 molar equivalents of TCEP (tris(2-carboxyethyl)phosphine) relative to the antibody were added from a stock solution in 10 mM water, and the antibody was reduced for 90 minutes. 8 molar equivalents of drug conjugate 2 were added from a stock solution in 10 mM DMA (dimethylacetamide), and the reduced antibody was coupled for 60 minutes. 8 molar equivalents of NAC (N-acetylcysteine) were added from a stock solution in 10 mM 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. Fractions containing protein were pooled and subjected to a final filtration through an appropriately sized 0.2 μm PES filter (Direct Chromatography / FIL-S-PES-022-13-100-S) under laminar flow at stage A. The final product was sampled for QC testing - monomer and [ADC] mg / ml by SEC-HPLC, mean DAR by PLRP, residual NMT inhibitor 1 by RP-HPLC, and endotoxin by Endosafe kinetic chromogenic assay.

[0729] ADC Example 2 - Preparation of Rituximab-NMT Inhibitor ADC (DAR 5)

[0730] A 10 mg / mL solution of rituximab was purchased. Prior to reduction and coupling, 5% v / v of 500 mM Tris, 25 mM EDTA, pH 8.5 was added to adjust the pH. 2.7 molar equivalents of TCEP (tris(2-carboxyethyl)phosphine) relative to the antibody were added from a stock solution in 10 mM water, and the antibody was reduced for 120 minutes. 8 molar equivalents of drug conjugate 2 were added from a stock solution in 10 mM DMA (dimethylacetamide), and the reduced antibody was coupled for 60 minutes. 8 molar equivalents of NAC (N-acetylcysteine) were added from a stock solution in 10 mM 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 30 mM histidine, 0.25 M sucrose 100 mM NaCl. Fractions containing protein were pooled and subjected to a final filtration through appropriately sized 0.2 μm PES filters (Direct Chromatography / FIL-S-PES-022-13-100-S) under laminar flow at stage A. The final product was sampled for QC testing - monomer and [ADC] mg / ml by SEC-HPLC, mean DAR by PLRP, residual NMT inhibitor 1 by RP-HPLC, and endotoxin by Endosafe kinetic chromogenic assay.

[0731] ADC Example 3 was prepared using the same method as described in ADC Example 1, except that saccharotuzumab was used instead of trastuzumab as the antibody.

[0732] ADC Example 4 was prepared using the same method as described in ADC Example 1, except that ifenatumab was used instead of trastuzumab as the antibody.

[0733] ADC Example 5 was prepared using the same method as ADC Example 1, except that drug conjugate 3 was used instead of drug conjugate 2.

[0734] Preparation of ADC Example 6

[0735] ADC Example 6 was prepared using drug conjugate 4. The preparation method of drug conjugate 4 was the same as that of drug conjugate 2, except that NMT inhibitor 21 was used instead of NMT inhibitor 1:

[0736]

[0737] Wherein steps 1 to 3 are as described for drug conjugate 2.

[0738] Herceptin (trastuzumab) was purchased and reconstituted to a 25.6 mg / mL solution. Prior to reduction and coupling, 5% v / v of 500 mM Tris, 25 mM EDTA (pH 8.5) was added to adjust the pH. 2.55 molar equivalents of TCEP (tris(2-carboxyethyl)phosphine) relative to the antibody was added from a 5 mM stock solution in water, and the antibody was left to reduce for 120 minutes. Prior to coupling, the reduced mAb was diluted 1 / 3 with PBS. 8 molar equivalents of drug conjugate 4 were added from a 10 mM stock solution in DMA (dimethylacetamide), and the reduced antibody was coupled for 90 minutes. 8 molar equivalents of NAC (N-acetylcysteine) were added from a 100 mM stock solution in water to quench unreacted drug conjugate 4, and allowed to react for 20 minutes. The conjugate was buffer exchanged into PBS using G25 resin (NAP25 column), then activated carbon was added in the ratio of 1mg carbon: 1mg ADC, and incubated overnight on a drum mixer at 10rpm at room temperature. The conjugate was then rotated 15 minutes at 4000xG to granulate the carbon, then the supernatant (ADC) was removed and filtered through a 0.2uM PES filter. The Amicon15 device was then used to further purify and concentrate the conjugate by diafiltration, wherein the PBS (pH 7.4) of 6x dialysis volumes (DV) was used in buffer exchange. The conjugate was then finally filtered through a 13mm 0.2μm PES filter (direct chromatography / FIL-S-PES-022-13-100-S) under a grade A laminar flow, and then 0.02% PS80 was prepared. The final product was sampled for QC testing - monomer and [ADC] mg / ml by SEC HPLC, mean DAR by PLRP, residual toxins by RP-HPLC, and endotoxins by Endosafe.

[0739] Preparation of ADC Example 7

[0740] ADC Example 7 was prepared using the same method as described in ADC Example 1, except that drug conjugate 5 was used, wherein the linker used was GGFG.

[0741] Structure of drug conjugate 5:

[0742]

[0743] It can be prepared using the same method as drug conjugate 2.

[0744] Preparation of ADC Example 8

[0745] Herceptin (trastuzumab) was purchased and reconstituted to a 25.6 mg / mL solution. Prior to reduction and coupling, 5% v / v of 500 mM Tris, 25 mM EDTA (pH 8.5) was added to adjust the pH. 2.55 molar equivalents of TCEP (tris(2-carboxyethyl)phosphine) relative to the antibody was added from a 5 mM stock solution in water, and the antibody was left to reduce for 120 minutes. Prior to coupling, the reduced mAb was diluted 1 / 3 with PBS. 8 molar equivalents of drug conjugate 1 were added from a 10 mM stock solution in DMA (dimethylacetamide), and the reduced antibody was coupled for 90 minutes. 8 molar equivalents of NAC (N-acetylcysteine) were added from a 100 mM stock solution in water to quench unreacted drug conjugate 1, and allowed to react for 20 minutes. The conjugate was buffer exchanged into PBS using G25 resin (NAP25 column), activated carbon was then added in the ratio of 1mg carbon: 1mg ADC, and incubated overnight on a drum mixer at 10rpm at room temperature. The conjugate was then rotated at 4000xG for 15 minutes to granulate the carbon, then the supernatant (ADC) was removed and filtered through a 0.2uM PES filter. The Amicon15 device was then used to further purify and concentrate the conjugate by diafiltration, wherein 6x dialysis volumes (DV) of PBS (pH 7.4) were used in buffer exchange. The conjugate was then finally filtered through a 13mm 0.2μm PES filter (direct chromatography / FIL-S-PES-022-13-100-S) under a grade A laminar flow, and then formulated into 0.02% PS80. The final product was sampled for QC testing - monomer and [ADC] mg / ml by SEC HPLC, mean DAR by HIC, residual toxins by RP-HPLC, and endotoxins by Endosafe.

[0746] Biological Example 1: HsNMT1 and SU-DHL-10 Assay

[0747] HSNMT1 IC 50

[0748] IC of some NMT inhibitors 50 The values ​​were measured using a sensitive fluorescence-based assay based on detection of CoA by 7-diethylamino-3-(4-maleimidophenyl)-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.

[0749] Cytotoxicity in SU-DHL-10 cell line

[0750] Certain NMT inhibitors were tested in the SU-DHL-10 cell line (human B-cell lymphoma). Compounds that show efficacy in this assay are expected to be useful as agents for treating or preventing hyperproliferative disorders such as cancer.

[0751] Cells were seeded in 96-well microplates and treated with compounds or cisplatin (as a positive control) at nine increasing concentrations in triplicate. The IC values ​​of test compounds and cisplatin were determined after 72 h of treatment in each cell line. 50 .

[0752] 1. On day 1, 90 μL of various cell suspensions (cell numbers ranging from 5,000 to 8,000 cells / well) were seeded into the wells of a 96-well plate (Corning). The number of cells to be seeded has been determined in advance.

[0753] 2. Place all 96-well plates containing cells in a 37°C, 5% CO2 incubator overnight.

[0754] 3. On day 2, observe the cells under a microscope to ensure that the cells treated with vehicle control are in good condition.

[0755] 4. Prepare a dilution series of test compounds and cisplatin at 10 times the desired final concentration. Add 10 μL / well of 10X compound solution to the corresponding plates. The final volume for all plates is 100 μL / well. The final DMSO concentration is 0.1%.

[0756] 5. On day 5 (after 72 hours of incubation), add 50 μL of CTG reagent to each well.

[0757] 6. Mix the contents on an orbital shaker for 5 minutes to promote cell lysis.

[0758] 7. Incubate the plate at room temperature for 10 minutes to stabilize the luminescent signal.

[0759] Luminescence was recorded using an EnVision multi-label microplate reader and data were analyzed using GraphPad Prism 8.0.

[0760] To calculate IC 50 , concentration response curves were generated using a nonlinear regression model with a sigmoidal concentration response. The formula used to calculate the % of viable cells is shown below, IC 50 Automatically generated by GraphPad Prism 8.0.

[0761]

[0762] (Lum Test Article = luminescence in test article treated wells; Lum Untreated = luminescence in vehicle treated wells; Lum Medium Control = luminescence in wells containing medium only without cells; Lum Untreated - Lum Medium Control was set as 100%).

[0763] result

[0764] The results of the above HsNMT1 and SU-DHL-10 experiments are shown in Table 2 below:

[0765] Table 2: Results of Biological Example 1

[0766] NMT inhibitors <![CDATA[HsNMT1 IC 50 (nM)]]> <![CDATA[SU-DHL-10IC 50 (nM)]]> 1 2.1 0.6 2 2.2 15.3 3 - - 4 - 9.4 5 - 12 6 2.2 5.9 7 - 5.0 8 9 - 9 - 0.7 10 - 0.6 11 - 2.0 12 - 0.9 13 - 0.7 14 - 0.4 15 - 1.2 16 - 0.2 17 2.0 1.0 18 1.9 5.3 19 - - 20 2.0 12.0 21 - 0.4 22 3.0 9 23 - - 24 - 0.7 25 2.0 1.0

[0767] The results of Biological Example 1 indicate that the tested NMT inhibitor compounds are inhibitors of HsNMT1 and exhibit potent in vitro cytotoxic activity.

[0768] Biological Example 2: Orthotopic Breast Cancer Xenograft Model

[0769] One day before tumor inoculation, NOD / SCID mice were implanted with an estrogen pellet (17β-estradiol, 60-day release, 0.36 mg) subcutaneously in the right flank. Then, on day -8, each mouse was inoculated with 1×10 7 Live BT474 breast cancer cells were suspended in 0.2 mL of a mixture of phosphate buffered saline and matrigel (1:1). The average tumor volume on day 0 of the study was 149.78 mm 3 At the time of treatment, the mice were assigned to the treatment group. Dosing began on the second day, and all animals were intravenously injected with trastuzumab, NMT inhibitor 1 or ADC Example 1. The study was terminated on the 35th day of the study. Mice were dosed once a week for four consecutive weeks, using vehicle alone (Group 1), 2.5mg / Kg trastuzumab (Group 2), 5mg / Kg trastuzumab (Group 3), 2.5mg / Kg ADC Example 1 (Group 4) or 5mg / Kg ADC Example 1 (Group 5), or 2mg / Kg NMT inhibitor 1, for 2 consecutive days, followed by a 5-day rest (Group 6). Each group consisted of 10 mice. The tumor volume of the mice was measured three times a week and the formula 0.5(LxW 2 ) to calculate tumor volume. Figure 1 and Figure 2The mean tumor volume (+SEM) for each study group at each measurement is shown, plotted as the last observation carried forward. Statistical analysis of tumor readings for Groups 1, 2, 3, 4, and 5 was performed using two-way ANOVA up to study day 23 (after which more than 50% of the animals were lost in one of the study groups; Group 2), or a Mixed-Effects model (PRISM GraphPad Software Inc.) was fitted when values ​​were missing in a group. Statistical analysis of Group 6 was performed until Day 13 (at which time the study was terminated due to significant weight loss observed). One mouse in Group 5 was determined to be an outlier (at the 5% confidence level) at all time points using ROUT outlier analysis in GraphPad Prism and was therefore excluded from the analysis.

[0770] ΔTGI% = ((C) mean - (C0) mean)) - ((T) mean - (T0) mean)) / ((C) mean - (C0) mean) x 100%, where T is the mean tumor volume of the treatment group on the measurement day, T0 is the mean tumor volume on study day 0. C is the mean tumor volume of the control group 1 mice on the measurement day, and C0 is the mean tumor volume on study day 0.

[0771] During the study, body weights of all animals were measured three times per week. Animals were given Diet Gel throughout the study. Figure 3 and Figure 4 Mean body weights for each group during the dosing period are presented.

[0772] result

[0773] The results of the treatment with trastuzumab alone or ADC Example 1 on tumor size are shown in Table 3 as a percentage of tumor growth inhibition. Figure 1 and Figure 2 Tumor volume (mm 3 )express. Figure 1 and Figure 2 Tumor volumes in mice treated with NMT inhibitor 1 alone were also depicted, but this study was terminated early due to significant weight loss observed in the treatment group.

[0774] Table 3: Tumor Growth Inhibition

[0775]

[0776] Mice treated with ADC Example 12.5 mg / kg (Group 4) had significantly reduced tumor volume compared to animals treated with trastuzumab 2.5 mg / kg (Group 2; p<0.0001) ( Figure 1In addition, the tumor volume of animals treated with ADC Example 1 5.0 mg / kg (Group 5) was significantly reduced compared to animals treated with trastuzumab 5 mg / kg (Group 3; p<0.0001) ( Figure 2 ). The tumor volume of mice treated with ADC Example 1 5.0 mg / kg (Group 5) was significantly reduced compared to animals treated with ADC Example 1 2.5 mg / kg (Group 4; p<0.0001) (see Table 3). The tumor volume of mice treated with ADC Example 1 5.0 mg / kg (Group 5) was significantly reduced compared to mice treated with 2.0 mg / Kg NMT inhibitor 1. The tumor size of mice treated with NMT inhibitor 1 2.0 mg / kg (Group 6) was reduced (see Table 3). Figure 1 and 2 ), but the study was stopped on day 13 due to significant weight loss observed in this study group. ADC Example 1 at a dose of 2.5 mg / kg was compared with NMT inhibitor 1 (Group 4, see Figure 1 ) have roughly the same effect, but the ADC Example 1 with a dose of 5.0 mg / kg has a better effect ( Figure 2 ). Administration of ADC Example 1 at a dose of 2.5 mg / kg delivered a dose of NMT Inhibitor 1 that was approximately 100-fold lower than NMT Inhibitor 1 administered alone, which means that ADC Example 1 was approximately 100-fold more potent than NMT Inhibitor 1 in vivo.

[0777] Figure 3 and Figure 4 The effect of treatment with trastuzumab alone, NMT inhibitor 1 alone, or ADC Example 1 on mouse body weight is shown. Mice treated with trastuzumab or ADC Example 1 were not significantly different from vehicle controls, while mice in Group 6 (NMT inhibitor 1) had a significant decrease in body weight ( Figure 3 and Figure 4 ), and the study was terminated early due to significant weight loss.

[0778] Biological Example 3: Gastric Cancer Xenograft Model

[0779] The purpose of this study was to preclinically evaluate the in vivo therapeutic efficacy of an antibody drug conjugate (ADC Example 1) in treating the subcutaneous NCI-N87 human gastric xenograft model in female BALB / c nude mice.

[0780] In this study, 143 mice were inoculated subcutaneously on the right flank with 1×10 7Live NCI-N87 tumor cells were suspended in 0.1 mL of a 1:1 mixture of PBS and Matrigel to promote tumor development. On day 0 of the study, 102 mice were assigned to nine treatment groups, and the average tumor volume was 168.08 mm 3 . Dosing began the next day, and animals were intravenously injected with vehicle control, trastuzumab, ADC Example 1, de trastuzumab, and isotype control antibody (isotype control) coupled to NMT inhibitor 1. The study was terminated on day 28 of the study. Mice were dosed once a week for two weeks, and were given 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 de trastuzumab (Group 6), 5 mg / Kg de trastuzumab (Group 7), or 5 mg / Kg isotype control antibody (Group 8).

[0781] Table 4: Summary of dosing regimens

[0782]

[0783]

[0784] The tumor volume of mice was measured three times a week and calculated using the formula 0.5(LxW2). Figure 5 and Figure 6 The mean tumor volume (+SEM) for each study group at each time of measurement is shown.

[0785] ΔTGI% = ((C) mean - (C0) mean)) - ((T) mean - (T0) mean)) / ((C) mean - (C0) mean) x 100%, where T is the mean tumor volume of the treatment group on the measurement day, T0 is the mean tumor volume on study day 0. C is the mean tumor volume of the control group 1 mice on the measurement day, and C0 is the mean tumor volume on study day 0.

[0786] During the study, body weights of all animals were measured three times per week. Animals were given Diet Gel throughout the study. Fig. 7A The mean body weights for each group during the dosing period are presented in Tables 7A (2.5 mpk study) and 7B (5 mpk study).

[0787] result

[0788] In Group 1 (vehicle control), 3 / 10 animals experienced significant weight loss (>10%); in Group 2 (2.5 mg / Kg trastuzumab), 1 animal experienced significant weight loss; in Group 3 (5 mg / Kg trastuzumab), 1 animal experienced significant weight loss; all mice regained weight at the next measurement. No significant weight loss was observed in any other group.

[0789] Compared with vehicle alone (Group 1), the tumor volume of mice treated with trastuzumab, ADC Example 1 and de trastuzumab at all concentrations (2.5 mg / kg or 5 mg / kg) was significantly reduced (p<0.0001). There was no significant difference in the isotype control ADC (Group 8; p=0.7935) compared with vehicle alone (Group 1).

[0790] Mice treated with trastuzumab 5 mg / kg (Group 3; p<0.0001) and ADC Example 1 5 mg / kg (Group 5; p<0.0001) had significantly reduced tumor volume compared to mice treated with trastuzumab 2.5 mg / kg (Group 2). Mice treated with de trastuzumab 2.5 mg / kg (Group 6; p<0.0001) and isotype control-ADC (Group 8; p<0.0001) had significantly higher tumor volume compared to Group 2 (trastuzumab 2.5 mg / kg). There was no significant difference between ADC Example 1 2.5 mg / kg (Group 4; p=0.8757) and de trastuzumab 5 mg / kg (Group 7; p=0.9965) compared to trastuzumab 2.5 mg / kg (Group 2).

[0791] Mice treated with ADC Example 1 5 mg / kg (Group 5; p<0.0001) had significantly reduced tumor volume compared to mice treated with trastuzumab 5 mg / kg (Group 3). There was no significant difference between Group 3 (Trastuzumab 5 mg / Kg) and Group 4 (ADC Example 1 2.5 mg / kg). In addition, the tumor volume of all other groups (Groups 6-8) was significantly larger than that of Group 3 (Trastuzumab 5 mg / kg).

[0792] Mice treated with ADC Example 1 5 mg / kg (Group 5; p<0.0001) had significantly reduced tumor volume compared to mice treated with ADC Example 1 2.5 mg / kg (Group 4). There was no significant difference between Group 4 and Group 7 (detrastuzumab 5 mg / kg, p=0.9932). In addition, the tumor volume of all other groups (Groups 6 and 8) was significantly higher than that of Group 4 (ADC Example 1 2.5 mg / kg).

[0793] The tumor volume of mice in Group 5 treated with ADC Example 1 5 mg / kg was significantly lower compared to all other groups (p<0.0001).

[0794] The tumor volume of mice treated with 5 mg / kg of trastuzumab (Group 7, p<0.0001) was significantly reduced compared to 2.5 mg / kg of trastuzumab (Group 6). The tumor volume of Group 8 was significantly larger compared to Group 6 (p<0.0001).

[0795] The tumor volume of mice treated with isotype control-ADC (Group 8; p<0.0001) was significantly higher than that of Group 7 (de-trastuzumab 5 mg / kg).

[0796] Compared to the vehicle group (Group 1), tumor growth inhibition (TGI) was significantly increased in all treatment groups except Group 8. When comparing tumor growth inhibition in all treatment groups, treatment with 5.0 mg / kg ADC Example 1 was the most effective (Group 5; TGI = 224.01%).

[0797] TGI% = ((C) mean - (C0) mean)) - ((T) mean - (T0) mean)) / ((C) mean - (C0) mean) x 100%, where T is the mean tumor volume of the treatment group on the measurement day, T0 is the mean tumor volume on study day 0. C is the mean tumor volume of the control group 1 mice on the measurement day, and C0 is the mean tumor volume on study day 0.

[0798] Biological Example 4: In vitro evaluation of trastuzumab coupled to NMT inhibitor 1 against HER2-positive breast cancer cells Cytotoxicity of cell line BT474

[0799] Trastuzumab is coupled to NMT inhibitor 1 (ADC Example 1) and tested in vitro for its cytotoxic activity against HER2-positive breast cancer cell line BT474. Cells were seeded in 96-well plates with a fusion rate of 10% (8000 for BT474). The next day, cells were treated with 50nM, 3.3nM and 0.2nM ADC Example 1 and controls in a medium containing 250nM Sytox Green. The plates were imaged every four hours in IncuCyte S3 for ten days. Images of % phase (fusion) and green (Sytox Green) were collected and analyzed using IncuCyte software 2022A. When cells were in logarithmic phase proliferation, data were expressed as phase area (μm) over time. 2 ) is the normalized green area.

[0800] Figures 8 to 11It was shown that ADC Example 1 effectively induced cell killing of BT474 cells at a concentration of 0.2 nM, while naked trastuzumab and isotype control IgG conjugated to NMT inhibitor 1 were unable to kill BT474 cells under these conditions. Trastuzumab was also effective in killing BT474 cells, but its effect was not as good as ADC Example 1 under these conditions. Puromycin was included as a positive cytotoxicity control.

[0801] Biological Example 5: In vitro evaluation of trastuzumab coupled to NMT inhibitor 1 against HER2-negative breast cancer cells Cytotoxicity of cell line MCF7

[0802] Trastuzumab is coupled to NMT inhibitor 1 (ADC Example 1) and tested in vitro for its cytotoxic activity against HER2-negative breast cancer cell line MCF7. Cells were seeded in 96-well plates with a fusion rate of 10% (MCF7 is 2000). The next day, cells were treated with 50nM, 3.3nM and 0.2nM ADC Example 1 and controls in a medium containing 250nM Sytox Green. The plates were imaged every four hours in IncuCyte S3 for ten days. Images of % phase (fusion) and green (Sytox Green) were collected and analyzed using IncuCyte software 2022A. When cells were in logarithmic phase proliferation, data were expressed as phase area (μm) over time. 2 ) is the normalized green area.

[0803] Figures 12 to 15 It was shown that ADC Example 1, isotype control IgG coupled to NMT inhibitor 1, trastuzumab or de-trastuzumab were unable 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.

[0804] Biological Example 6: In vitro "bystander effect" of trastuzumab coupled to NMT inhibitor 1

[0805] HER2 negative MCF7 cells were stably transduced with pHIV eGFP (Addgene plasmid 21373) and sorted by flow cytometry to obtain a cell population of 100% GFP positive (green fluorescent protein). After two passages, eGFP positive MCF7 cells were seeded in 96-well plates with a ratio of 1:1 with wild-type MCF7 cells (1000 cells of each kind) or HER2 positive BT474 cells. In culture medium (50% MCF7 and 50% BT474 cell culture medium), cells were treated with ADC Example 1, Reference Example 1 (trastuzumab-monomethyl ruiocetine E (MMAE)), isotype control (defined as described above), detrastuzumab or naked trastuzumab (concentration of 12.5nM, 3.13nM, 0.78nM, 0.2nM and 0.05nM) and control. Cells were imaged once every four hours in IncuCyte S3 for ten days. Images of % green (eGFP+MCF7) were collected and analyzed using IncuCyte software 2022A. Data are presented as % green area (μm 2 ) is indicated as a measure of eGFP+MCF7 confluency at day 9.

[0806] Fig.16 The results showed that when eGFP+MCF7 cells were co-cultured with wild-type MCF7 cells, no significant cytotoxicity occurred when incubated with ADC Example 1 for 9 days, nor did significant cytotoxicity occur with any control or reference standard. This is expected because trastuzumab targets HER2-positive cells. However, when eGFP+MCF7 cells were co-cultured with HER2-positive BT474 cells ( Fig.17 ), GFP signal was significantly lost, which corresponds to bystander killing of eGFP+MCF7 cells. Among the three ADCs tested (ADC Example 1, Reference Example 1, and trastuzumab), ADC Example 1 was the most effective in inducing bystander killing, ADC Reference Example 1 was the second most effective, and trastuzumab was the least effective. Naked trastuzumab and isotype control IgG coupled to NMT inhibitor 1 were used as controls, and neither induced any bystander killing.

[0807] Biological Example 7: In vitro evaluation of cytotoxicity of antibodies conjugated to NMT inhibitor 1

[0808] NMT inhibitor 1 was conjugated to trastuzumab (ADC Example 1) and to tacitrullumab (ADC Example 3) as described. NMT inhibitor 26 was conjugated to trastuzumab (ADC Example 5). ADC Examples 1, 3, and 5 were tested for 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 to 24 ).

[0809] NMT inhibitor 1 was conjugated with ifenatumomab as described (ADC Example 4) and tested for its cytotoxic activity against a panel of prostate cancer cell lines including LNCaP, C42 and VCaP ( Figures 25 to 27 ). The cells were seeded in a suitable growth medium in a 96-well flat-bottom plate at a set initial density. After 24 hours, serial dilutions of each test compound were added, with concentrations ranging from 10nM to 0.001nM of NMT inhibitor 1, or 50nM to 0.005nM of various ADCs. After 144 hours of exposure, cell proliferation was measured by CellTiter-Glo2.0 solution cell viability assay. The percentage inhibition of viability relative to the DMSO vehicle control at the endpoint was calculated using the following formula (after blank correction, time zero correction, and logarithmic transformation of compound concentration):

[0810] Viability % = (luminescence of treated cells on day 6 - blank) - (luminescence on day 0 - blank) / (average luminescence of vehicle-treated cells on day 6 - blank) - (luminescence on day 0 - blank)

[0811] result

[0812] Table 5 shows the inhibition of cell viability quantified as described above, defined as either absolute IC50 (the concentration of inhibitor at which the response is reduced by half) or relative IC50 (the concentration required to bring the curve down to a position midway between the top and bottom plateaus of the curve). Figures 18 to 27 The complete concentration response curves (mean + / - SEM) for each cell line are shown.

[0813] Table 5: Inhibition of cell viability

[0814]

[0815] Biological Example 8: Evaluation of test samples using the CellTiter-Glo (CTG) luminescent cell viability assay as Effects of single agents on the viability of gastric cancer organoid cells

[0816] method

[0817] Day 1: Patient-derived gastric cancer organoids (PDXO) were sheared into uniform size and the desired number of organoids were mixed with 50% Matrigel at a 1:1 ratio to generate organoids of appropriate size for screening.

[0818] Day 0: Organoid seeding

[0819] 1. Collect organoids from each well of a 6-well plate by adding 20 μl of 100x Dispase solution to each well of the plate containing 2 ml of organoid medium.

[0820] 2. Return the plate to the incubator and incubate at 37°C for 30 minutes.

[0821] 3. Collect organoids from all wells and pipette through a pre-wetted 100 μm filter into a 50 ml plastic tube.

[0822] 4. After all wells have been filtered through a 100 μm filter, filter the flow-through through a pre-wet 20 μm filter.

[0823] 5. Invert the 20 μm filter and recover the organoids in a new 50 ml tube.

[0824] 6. Collect the organoids and resuspend them in the appropriate culture medium. Count the organoids to obtain the concentration.

[0825] 7. Use culture medium to adjust the cell concentration to an appropriate concentration.

[0826] 8. Add Matrigel to a final concentration of 5% v / v and place the organoid suspension on ice.

[0827] 9. Use a Multidrop dispenser to add 40 μL of cell suspension and corresponding culture medium to a 384-well plate at the following seeding densities:

[0828]

[0829]

[0830] 10. Return the screening plate to the incubator before adding compounds and taking the Day 0 reading. Two replicate plates are set up. One for the Day 0 reading (T0) and the other for the endpoint reading.

[0831] Day 0: Compound treatment and day 0 baseline CTG (cell titre glo) readings

[0832] 11. Add test samples according to the drug dilution scheme and plate map through Tecan D300e

[0833] 12. Return the screening plate to the incubator.

[0834] 13. For Day 0 read plates, add 40 μL CTG 3D per well and read luminescent signal on Envision plate reader.

[0835] Day 6: Endpoint CTG reading.

[0836] 1. Read the luminescent CTG signal on the assay plate at the end of the assay: add 40 μL CTG 3D to each well via a Multidrop dispenser, mix the contents on a plate shaker for 5 minutes, and then incubate the plate for 30 minutes at room temperature in the dark. Read the luminescent signal on an Envision plate reader.

[0837] Data were displayed graphically using GraphPad Prism. To calculate absolute IC50, concentration-response curves were fitted using a nonlinear regression model with a sigmoidal dose response with variable slope. The formula for calculating survival is shown below, and absolute IC50 was calculated from the dose response curve generated by GraphPad Prism.

[0838] Survival rate (%) = (Lum test article - Lum medium control) / (Lum vehicle control - Lum medium control) x 100%.

[0839] Gastric cancer organoids were incubated for 5 or 6 days with a range of concentrations of (1) isotype control ADC, (2) ADC Example 1, (3) staurosporine as a positive control, (4) trastuzumab or (5) de-trastuzumab (trastuzumab-DXd). Table 6 shows the IC50 (μM), maximum inhibition (%) and area under the curve.

[0840] Table 6: Results of organoid studies

[0841]

[0842]

[0843]

[0844] Concentration response curves of 16 gastric cancer organoids Figures 28 to 43 Data show organoid viability (mean + / - SEM) after treatment with isotype control, ADC Example 1, de-trastuzumab (DXd), and trastuzumab conjugated to NMT inhibitor 1.

[0845] The results showed that in most gastric cancer organoids, ADC Example 1 was the most effective, as shown by its IC50 value compared to the control. In some organoids, ADC Example 1 had comparable activity to trastuzumab-DXd, as shown by the IC50 value.

[0846] Biological Example 9: Maximum Tolerated Dose Study Following a Single Intravenous Injection of ADC Example 1 in Male Cynomolgus Monkeys

[0847] Six routine, non-naive male cynomolgus macaques were used in this MTD study. The service provider laboratory that conducted this study has been certified by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC). The animal care and use application for this study was submitted to the service provider's Institutional Animal Care and Use Committee (IACUC) for approval. The IACUC reviewed this protocol for compliance with its TSPIACUC policies and procedures. Monkeys were quarantined at the supplier for at least 2 weeks before being shipped to the service provider's facility. Animals were screened for active infection with tuberculosis, herpes B, simian acquired immunodeficiency virus (SIV), simian retrovirus (SRV), simian T-cell lymphotropic virus (STLV), Shigella, and Salmonella, and their normal hematology and clinical chemistry parameters were assessed. Serology or stool analysis was performed to look for evidence of parasites.

[0848] The animals were randomly divided into 3 groups with 2 animals in each group and the test article ADC Example 1 was administered by slow intravenous injection (5 minutes) according to Table 7.

[0849] Table 7: Dosage regimen for Biological Example 9

[0850]

[0851]

[0852] Animals were evaluated during survival according to Table 8.

[0853] Table 8: Evaluation scheme for Biological Example 9

[0854]

[0855] Blood samples were collected from each study animal on the following days: one day before dosing (Day 1), on the second day of the study, and at termination (Day 7 or Day 11). Animals were fasted overnight before samples were collected for clinical pathology analysis. Blood samples were collected by puncturing peripheral blood vessels. Blood samples were processed and analyzed for hematology, clinical biochemistry, and coagulation. The 5, 10, and 20 mg / Kg doses were well tolerated and no significant changes occurred in hematology or clinical chemistry parameters. In particular, the 5 and 10 mg / Kg groups did not show any changes. Fig.44a and 44b Representative data for the 20 mg / Kg group are shown. Specifically, the mean (+ / -SEM) levels of hematological markers are plotted as normalized values ​​for baseline (pre-dose) values. No changes were observed on Day 7, while the changes observed on Day 11 were relatively small and not significant.

[0856] Biological Example 10: In vivo tolerance study of ADC Example 1 in non-tumor bearing female CD1 mice

[0857] The purpose of this study was to evaluate the in vivo tolerability of ADC Example 1 in female CD1 mice. Body weight was measured daily, and terminal whole blood and serum were collected from each animal at the end to measure complete blood counts and blood biochemistry parameters.

[0858] 24 mice were randomly divided into 4 groups and dosing began on the second day. All animals were injected intravenously with a single dose of 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 a complete autopsy was performed on all animals and 2 naive animals at the end of the study.

[0859] Significant weight loss (>10%) was observed in 3 animals of Group 4 (ADC Example 1 100 mg / kg) on ​​study day 5, and Group 4 was terminated on study day 6 ( Fig.45 No significant weight loss was observed in any other group ( Fig.45 ), and no clinical observations were observed in any of the animals in the study.

[0860] At the end of the study, whole blood was collected from all animals for complete blood count (CBC), and serum was collected for blood biochemistry testing. Fig.46 The mean values ​​of AST (aspartate aminotransferase), ALT (alanine aminotransferase), ALP (alkaline phosphatase), LDH (lactate dehydrogenase), CK (creatine kinase) and GGT (γ-glutamyl transferase) in the blood of animals at the end of the experiment are shown. Only group 4 (100 mg / Kg) showed significant changes. Fig.47 The mean values ​​of cells in the blood of animals at termination are shown: LYM (lymphocytes), MON (monocytes), NEU (neutrophils), RBC (red blood cells), HGB (hemoglobin) and PLT (platelets). Significant changes were observed only in Group 4 (ADC Example 1 100 mg / Kg).

[0861] In conclusion, ADC Example 1 was well tolerated in mice after a single IV dose of up to 50 mg / Kg, which is 10 times the effective dose observed in Biological Examples 2 and 3.

[0862] Biological Example 11: LNCaP prostate cancer xenograft model.

[0863] The purpose of this study was to evaluate the efficacy of ADC Example 4 in male NOD SCID mice bearing LNCaP tumors.

[0864] A total of 84 male NOD SCID mice, 5-8 weeks old and weighing 25-30 g, were used for the study. 7LNCaP tumor cells (viability 78%, confluence approximately 70-80%) were implanted subcutaneously into the flank of male NOD SCID mice. 3 At 4 pm, animals were assigned to treatment groups as shown in Table 9 below, with 10 mice assigned to each group, each group having similar tumor volume means and distribution. Mice were treated with vehicle alone, unconjugated ifinatamab, ifinatamab-deruxtecan (ifinatamab-DXd), or ADC Example 4.

[0865] Table 9: Dosing regimen for Biological Example 11

[0866]

[0867] Observation period: 35 days.

[0868] Dosage: All IV doses are 5 mL / kg

[0869] During the study, no adverse reactions were observed at any dose, and the mean body weight in each group remained within 10% of the pre-treatment level ( Fig.49 ).

[0870] Individual cases of weight loss >10% were observed at various time points in the study. Three weeks after the first animal was treated, all mice were given DietGel to mitigate weight loss. No animals were euthanized prematurely due to weight loss, and these instances of weight loss were likely related to tumor burden.

[0871] On study day 28, animals receiving three Q7D doses of 10 mg / kg of ADC Example 4 exhibited significantly higher mean body weights than animals receiving vehicle control treatment (one-way ANOVA, Dunnett's p=0.0046). Body weights of the other treatment groups were not significantly different from the vehicle group at this time.

[0872] Two animals were euthanized prematurely due to welfare concerns. The first animal was euthanized on day 26 of treatment with 10 mg / kg ifenatumab, while the second animal was euthanized on day 33 of treatment with 5 mg / kg ifenatumab-DXd. Both animals were euthanized primarily due to respiratory dysmotility. Large spontaneous thymic tumors were documented at necropsy in each animal.

[0873] During the study, tumors in the vehicle-treated group grew steadily, reaching a mean volume of 752 ± 89.4 mm on study day 28. 3 .

[0874] On day 28, treatment with 10 mg / kg or 5 mg / kg of ifinatumomab had no significant effect on LNCaP tumor volume, and the tumor growth curves of animals receiving this therapy were generally similar to those of animals treated with vehicle alone ( Fig.48 , Table 10).

[0875] Treatment with 10 mg / kg ifenatumab-DXd significantly reduced the mean volume of LNCaP tumors by day 28 compared with vehicle controls. Tumor growth rates were mostly lower in animals receiving this therapy than in control animals. Ifenatumab-DXd at a dose of 5 mg / kg slowed the growth of LNCaP tumors to a lesser extent ( Fig.48 , Table 10).

[0876] All animals dosed with 10 mg / kg ADC Example 4 showed tumor regression within three weeks after the start of treatment ( Fig.48 , Table 10), the therapy significantly reduced mean tumor volume (TV) compared to vehicle starting on day 7 (Mann-Whitney test). By day 28, each tumor had regressed to ≤25% of its volume at the start of treatment.

[0877] Likewise, animals receiving 5 mg / kg ADC Example 4 had significantly reduced tumor volumes from day 7 to day 28 compared to the control group (Mann-Whitney test). By day 28, tumor volumes in all but one animal were below those recorded at the start of treatment.

[0878] Table 10. Comparison of tumor volumes among treatment groups.

[0879] Adjusted p values ​​relative to vehicle control were calculated by Kruskal-Wallis test and Dunn's multiple comparisons. TGI% = ((C) mean - (C0) mean)) - ((T) mean - (T0) mean)) / ((C) mean - (C0) mean) x 100%, where T is the mean tumor volume of the treatment group on the measurement day, and T0 is the mean tumor volume on study day 0. C is the mean tumor volume of mice in control group 1 on the measurement day, and C0 is the mean tumor volume on study day 0.

[0880]

[0881] Biological Example 12: VCaP prostate cancer xenograft model.

[0882] The purpose of this study was to preclinically evaluate the in vivo therapeutic efficacy of ADC Example 4 in treating the subcutaneous VCaP human prostate cancer xenograft model in intact male CB17 / SCID mice.

[0883] In this study, 144 mice were inoculated subcutaneously in the right anterior flank area with 1x10 7 The live VCaP tumor cells were suspended in 0.1 mL of PBS and Matrigel (1:1) mixture for tumor development. The average tumor volume on day 0 of the study was about 162.16 mm 3 80 mice were assigned to 8 treatment groups. Dosing began the next day, and all animals were intravenously administered ADC Example 4, ifenatumab-Drunotecan (ifenatumab-Dxd) or uncoupled ifenatumab. All mice received two doses of test agents, respectively on the first day of the study and the eighth day of the study. The study was terminated on the 30th day of the study.

[0884] The 8 groups are allocated as follows:

[0885] Group 1 Vehicle Control

[0886] Group 2 Unconjugated ifenatumomab 5 mpk

[0887] Group 3 Unconjugated ifenatumomab 2.5 mpk

[0888] Group 4: Ifenatumomab-Drugtec 5 mpk

[0889] Group 5: Ifenatumomab-Drunotecan 2.5 mpk

[0890] Group 6 ADC Example 4 10mpk

[0891] Group 7 ADC Example 4 5mpk

[0892] Group 8 ADC Example 4 2.5mpk

[0893] No significant weight loss was observed in any of the animals studied ( Fig.51 ).

[0894] When compared to vehicle alone (Group 1), 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) had significantly reduced tumor volume (p<0.0001). Ifinacomab-Dxd 5 mg / kg (Group 4; p=0.0078) showed significantly larger tumor volume compared to vehicle alone (Group 1), while Ifinacomab-Dxd 2.5 mg / kg (Group 5; p=0.8127) and both concentrations of unconjugated Ifinacomab (Group 2, p=0.1104; and Group 3, p=0.6703) were not significantly different from vehicle alone (Group 1), see Fig.50 .

[0895] 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) had significantly reduced tumor volume compared to all other treatment groups (Groups 2-5; p<0.0001). There was also evidence of a dose response, with Group 6 (10 mg / kg) having the greatest reduction in tumor volume, followed by Group 7 (5 mg / kg), and then Group 8 (2.5 mg / kg); all groups were significantly different (p<0.0001), see Fig.50 .

[0896] The tumor volume of mice treated with ifenatumomab-Dxd 2.5 mg / kg (Group 5) was not significantly different from that of mice in Groups 2, 3, and 4 (unconjugated ifenatumomab 5 mg / kg, unconjugated ifenatumomab 2.5 mg / kg, ifenatumomab-Dxd 5 mg / kg, respectively). The tumor volume of mice treated with ifenatumomab-Dxd 5 mg / kg (Group 4) was significantly higher than that of mice treated with unconjugated ifenatumomab 5 mg / kg (Group 2, p=0.0002) and unconjugated ifenatumomab 2.5 mg / kg (Group 3, p=0.0216), see Fig.50 .

[0897] Compared with the vehicle group (Group 1), all treatment groups had significantly increased tumor growth inhibition (ΔTGI). When comparing tumor growth inhibition of all treatment groups, treatment with 10 mg / kg and 5 mg / kg ADC Example 4 was the most effective (Group 6, ΔTGI = 114.03%; and Group 7, ΔTGI = 114.69%), see Table 11.

[0898] Table 11. Dosage regimen and results of Biological Example 12

[0899]

[0900] Biological Example 13: JIMT-1 breast cancer xenograft model.

[0901] The purpose of this study was to preclinically evaluate the in vivo therapeutic efficacy of ADC Example 3 in the subcutaneous JIMT-1 human breast xenograft model in female NOD / SCID mice.

[0902] In this study, 128 mice were inoculated subcutaneously in the right anterior flank area with 5x10 6 The live JIMT-1 tumor cells were suspended in 0.1 mL PBS to promote tumor development. The average tumor volume on day 0 of the study was approximately 160.66 mm 3At 14:00, 80 mice were assigned to 8 treatment groups. Dosing began on the second day, and all animals were intravenously administered ADC Example 3, saxetuzumab-govitecan or unconjugated saxetuzumab. The study was terminated on the 60th day of the study. The 8 groups were assigned as follows:

[0903] Group 1 Vehicle Control

[0904] Group 2 Unconjugated tacizumab 5 mpk

[0905] Group 3 Unconjugated taciturnumab 2.5 mpk

[0906] Group 4 Sacilotuzumab 5 mpk (ADC Example 3, 5 mg / kg added on days 27 and 34)

[0907] Group 5 Sacilotuzumab 2.5 mpk (ADC Example 3, 5 mg / kg added on day 27)

[0908] Group 6 ADC Example 3 10mpk

[0909] Group 7 ADC Example 3 5mpk

[0910] Group 8 ADC Example 3 2.5mpk

[0911] Significant weight loss (>10%) was observed in one animal in Group 7. No significant weight loss was observed in any other group studied, see Fig.54 and 55 On study day 10, one animal in Group 2 was found dead.

[0912] 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) had significantly reduced tumor volume (p<0.0001) when compared to vehicle alone (Group 1). Both concentrations of taciturnin (Group 4, 5 mg / kg, p<0.0001; Group 5, 2.5 mg / kg, p=0.0348) resulted in significantly reduced tumor volume when compared to vehicle alone (Group 1). When compared to vehicle alone (Group 1), tumor volume was significantly reduced after the use of unconjugated taciturnin in Group 2 (5 mg / kg, p=0.0028), but there was no significant difference in Group 3 (2.5 mg / kg, p=0.0586). The data for the 5 mg / kg and 10 mg / kg groups are shown in Figure 2. Fig.52 The data of the 2.5 mg / kg group are shown in Fig.53 shown.

[0913] Compared with all other treatment groups (Groups 2-5; p<0.0001), the tumor volume of 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) was significantly reduced. There was also evidence of a dose response, with Groups 6 (10 mg / kg) and 7 (5 mg / kg) showing the greatest tumor volume reduction compared to Group 8 (2.5 mg / kg) (p<0.0001).

[0914] Although Group 4 Sacitotuzumab-Govitecan 5 mg / kg showed a significant reduction in tumor volume compared to unconjugated Sacitotuzumab Group 2 (5 mg / kg, p=0.0097) and Group 3 (2.5 mg / kg, p=0.0499), Group 5 (Sacitotuzumab-Govitecan, 2.5 mg / kg) was not significantly different from Group 2 (p=0.8424) and Group 3 (p=0.9995). There was also no significant difference between the two concentrations of unconjugated Sacitotuzumab in Group 2 (5 mg / kg) and Group 3 (2.5 mg / kg), p=0.9837. However, there was a significant difference between the two concentrations of Sacitotuzumab-Govitecan in Group 4 (5 mg / kg) and Group 5 (2.5 mg / kg), p=0.3802.

[0915] Compared with the vehicle group (Group 1), all treatment groups had significantly increased tumor growth inhibition (ΔTGI). When comparing tumor growth inhibition of all treatment groups, treatment with 10 mg / kg and 5 mg / kg ADC Example 3 was the most effective (Group 6, ΔTGI = 121.55%; and Group 7, ΔTGI = 122.04%).

[0916] Since treatment with sacizumab-govitecan was only partially effective in Group 4, additional doses of ADC Example 3 were administered at 5 mpk IV on study days 27 and 34. This resulted in a significant reduction in tumor volume compared to vehicle controls (see Fig.52 Group 5 also received one dose of ADC Example 3 (IV, 5 mpk) on study day 27 (see Fig.53 ), although no significant response was observed.

[0917] Table 12. Dosage regimen and results of Biological Example 13

[0918]

[0919]

[0920] Biological Example 14: In vitro evaluation of trastuzumab coupled to NMT inhibitor 1 against HER2-positive breast cancer cells Cytotoxicity of cell line BT474

[0921] Trastuzumab is coupled to NMT inhibitor 1 (ADC Example 8), and its cytotoxic activity against HER2-positive breast cancer cell line BT474 is tested in an in vitro assay. Cells are seeded in 96-well plates with a fusion rate of 10% (8000 for BT474). The next day, cells were treated with ADC Example 8 and controls at 50nM, 3.13nM and 0.2nM in a medium containing 250nM Sytox Green. The plates were imaged every four hours in IncuCyte S3 for ten days. Images of % phase (fusion) and green (Sytox Green) were collected and analyzed using IncuCyte software 2022A. When cells are in logarithmic phase proliferation, data are expressed as phase area (μm) over time. 2 ) is the normalized green area.

[0922] Fig.56 It was shown that ADC Example 8 effectively induced cell killing of BT474 cells at a concentration of 0.2 nM, while naked trastuzumab and isotype control IgG conjugated with NMT inhibitor 1 were unable to kill BT474 cells under these conditions (see Fig. 9 and 11 Trastuzumab was also effective in killing BT474 cells, but was less effective than ADC Example 8 under these conditions (see Fig.10 ). Puromycin was included as a positive cytotoxicity control.

[0923] Biological Example 15: In vitro evaluation of trastuzumab coupled to NMT inhibitor 1 against HER2-negative breast cancer cells Cytotoxicity of cell line MCF7

[0924] Trastuzumab is coupled to NMT inhibitor 1 (ADC Example 8), and its cytotoxic activity against HER2-negative breast cancer cell line MCF7 is tested in an in vitro assay. Cells are seeded in 96-well plates with a fusion rate of 10% (MCF7 is 2000). The next day, cells were treated with ADC Example 8 and controls at 50nM, 3.13nM and 0.2nM in a culture medium containing 250nM Sytox Green. The plates were imaged every four hours in IncuCyte S3 for ten days. Images of % phase (fusion) and green (Sytox Green) were collected and analyzed using IncuCyte software 2022A. When cells were in logarithmic phase proliferation, data were expressed as phase area (μm) over time. 2 ) is the normalized green area.

[0925] Fig.13 , 14, 15 and 57 show that ADC Example 8, isotype control IgG coupled to NMT inhibitor 1, trastuzumab or de-trastuzumab were unable to induce cell killing of HER2-negative MCF7 cells at concentrations up to 50 nM under these conditions. This is expected because trastuzumab targets HER2-positive cells. Puromycin was included as a positive cytotoxicity control.

[0926] in conclusion

[0927] The results of Biological Example 1 indicate that the tested NMT inhibitor compounds are inhibitors of HsNMT1 and exhibit potent in vitro cytotoxic activity.

[0928] The results of Biological Example 2 indicate that the ADC tested in the present invention exhibited potent in vivo cytotoxic activity and had improved in vivo tolerability when compared to NMT inhibitor 1.

[0929] The results of Biological Example 3 demonstrated that the test ADC of the present invention, i.e., ADC Example 1, exhibited better efficacy than other treatment groups in inhibiting tumor growth in a gastric cancer xenograft mouse model (see Table 4), and no significant weight loss was observed.

[0930] The results of Biological Example 4 show that the test ADC of the present invention, ADC Example 1, effectively induces cell killing of the HER2-positive breast cancer cell line BT474. Compared with other test compounds: trastuzumab, de-trastuzumab and isotype control, the ADC of the present invention is more effective in inducing cell killing.

[0931] The results of Biological Example 6 showed that in eGFP+MCF7 cells co-cultured with HER2-positive BT474 cells, ADC Example 1 was the most effective in inducing bystander killing compared to other ADCs and controls.

[0932] The results of Biological Example 7 demonstrate that various ADCs of the invention (particularly ADC Examples 1 and 3-5) have cytotoxic activity against a range of cancer cell lines, as measured by cell viability assays. Figures 18 to 27 As shown in the data, various ADCs of the present invention (particularly ADC Examples 1 and 3-5) have improved cytotoxicity compared to the corresponding controls.

[0933] 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 to the control. In some organoids, ADC Example 1 has activity comparable to that of trastuzumab-DXd, as shown by the IC50 value.

[0934] The results of Biological Example 9 showed that ADC Example 1 was well tolerated in the monkey model, with no significant changes in hematological or clinical chemistry parameters even at the highest dose of 20 mg / kg / day.

[0935] The results of Biological Example 10 show that ADC Example 1 was well tolerated after a single IV administration to mice at doses up to 50 mg / Kg, which is 10 times the effective dose observed in Biological Examples 2 and 3.

[0936] The results of Biological Example 11 show that in the LNCaP prostate cancer xenograft model, mice have good tolerance to both 5 mg / kg and 10 mg / kg doses of ADC Example 4. Animals receiving these doses of ADC Example 4 showed a significant reduction in tumor volume, while treatment with ifenatumab alone had no significant effect on tumor volume. ADC Example 4 also performed better than ifenatumab-DXd at both doses.

[0937] The results of Biological Example 12 show that in the VCaP human prostate cancer xenograft model, mice have good tolerance to ADC Example 4 at 2.5mg / kg, 5mg / kg and 10mg / kg doses. ADC Example 4 reduces tumor volume more than ifinatumomab-Dxd and unconjugated ifinatumomab. Indeed, there is no significant difference in the use of unconjugated ifinatumomab compared with the use of vehicle alone. Compared with the control, ADC Example 4 causes the most tumor growth inhibition, see Table 11.

[0938] The results of Biological Example 13 show that in the JIMT-1 human breast cancer xenograft model, mice were well tolerated at 2.5 mg / kg, 5 mg / kg and 10 mg / kg doses of ADC Example 3. Compared with all other treatment groups, the tumor volume of mice treated with all three concentrations of ADC Example 3 was significantly reduced, as shown by the ΔTGI results in Table 12, where ADC Example 3 caused the most tumor growth inhibition.

[0939] The results of Biological Example 14 show that the test ADC of the present invention, ADC Example 8, effectively induces cell killing of the HER2-positive breast cancer cell line BT474. Compared with other test compounds: trastuzumab, de-trastuzumab and isotype control, the ADC of the present invention is more effective in inducing cell killing.

[0940] Therefore, the ADCs of the present invention are promising for use as drugs in general, and in particular in the treatment of hyperproliferative disorders such as cancer.

[0941] Throughout the 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 stated 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.

[0942] All patents and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

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

2. A salt of the ADC according to claim 1. The salt according to claim 1 or 2, which is a pharmaceutically acceptable salt.

4. The ADC according to 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): or a salt thereof, in: Y is selected from -CH-, -C(R 2 )- and -N-; R 1 is a group of formula –X–L–A; X stands for -O-; L stands for -(CH2) m -; m is 1, 2 or 3; A is a 6-10 membered aromatic carbocyclic ring or a 5-10 membered aromatic heterocyclic ring, the aromatic carbocyclic ring or heterocyclic ring is optionally substituted by 1, 2 or 3 substituents, each substituent is independently selected from the group consisting of: -F, -Cl, -Br, -OCH3, -OCF3, -CN, optionally substituted by up to 3 halogens, hydroxyl or -OC 1-4 Alkyl-substituted-C 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)2, -CH2C(O)N(R 9 )2、-CH2C(O)N(R 13 )C 1-4 Alkyl OC 1-4 Alkyl, -CH2C(O)N(C 1-4 Alkyl OC 1-4 Alkyl)2, -S(O)2NHC 1-4 Alkyl, -S(O)2N(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, CH2N(R 13 )2、CH2N(R 13 )C(O)C 1-4 Alkyl, CH2N(R 13 )S(O)2C 1-4 Alkyl, -CH2S(O)2C 1-4 Alkyl and CO2H; s is 0, 1, 2, or 3; Each R 2 independently selected from the group consisting of: -F, -Cl, -Br, -OCH3, -OCF3, -CN, -C optionally substituted with up to 3 halogens 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)2N(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 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 C optionally substituted by up to 3 -F, -Cl, -Br, -OH, -OCH3, -OCF3 or -CN groups 1-6 alkyl; When present, R 10 is hydrogen or methyl; When present, R 11 is hydrogen or methyl; or R 3 Group and R 5 The groups and the atoms between them form bonds between the atoms between them, or between the atoms and -(CHR a ) r - a 3- to 7-membered non-aromatic heterocyclic ring; or R 10 Group and R 5 The groups and the atoms between them form a a ) r - a 3- to 7-membered non-aromatic heterocyclic ring; r is 1, 2, 3, 4 or 5; R a is hydrogen or methyl; Each R 7 are independently selected from hydrogen, halogen, C 1-4 Alkoxy and optionally substituted by 1, 2 or 3 halogen 1-4 alkyl; and R 8 Selected from hydrogen and C 1-4 alkyl; Each R 9 independently selected from hydrogen and C 1-4 Alkyl groups, or two R 9 The groups and the N to which they are bound form a 4- to 7-membered non-aromatic heterocyclic ring, the heterocyclic ring optionally containing 1 or 2 further heteroatoms selected from N, O and S; Each R 13 independently selected from hydrogen and C 1-4 alkyl; and wherein: i) E, J and G are each C(R 7 ), K is carbon, 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. The ADC or salt thereof according to claim 5, wherein the NMT inhibitor is a compound of formula (IA^^): or a salt thereof, in: R 1 is a group of formula -XLA; A is 4-pyrazolyl, which is optionally substituted with up to 3 substituents selected from methyl and -C(O)N(CH3)2; X is -O-; L is –(CH2) m -; m is 2; R 2′ Selected from the group consisting of fluorine or chlorine (preferably fluorine); R 2″ 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 The R 3 The group and the R 6 The groups and the atoms therebetween form a 3- to 7-membered non-aromatic heterocyclic ring consisting of the atoms and bonds therebetween; E, J, G, K, Q and M are: i) E, J and G are each CH, K is carbon, 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 not more than one -C(O)N(CH3)2 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): or a salt thereof, in: R 1 is H or -CH3; and R 2 It is H or F.

8. The ADC or a salt thereof according to claim 7, wherein 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: 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): or a salt thereof, in: n1 is 0, 1, 2, 3, 4, 5 or 6; Ring A* is an optionally substituted nitrogen-containing aryl group, wherein each substitutable carbon or nitrogen in Ring A* is optionally and independently replaced by one or more R 5A substituted, and wherein if ring A* contains a -NH- moiety, the nitrogen may be optionally replaced by C 1-6 alkyl (eg, methyl) substituted; and wherein R 4A and ring A* may form a cyclic group together with the atoms to which they are attached, Ring B* is an optionally substituted aryl or heteroaryl group, wherein each substitutable carbon or heteroatom in Ring B* is optionally and independently replaced by one or more R 3A replace; One of W and X may not be present, and W and X are independently selected from R 11A , optionally by R 11A Substituted hydrocarbon groups (e.g., C 1-8 alkyl, alkenyl, alkynyl or haloalkyl) and optionally R 12A Substituted -(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 are independently selected from hydrogen, R 12A , optionally by R 12A Substituted hydrocarbon groups (e.g., C 1-6 alkyl, alkenyl, alkynyl or haloalkyl) and optionally substituted by one or more R 12A Substituted -(CH2) L1 -heterocyclic group; wherein R 2A Together with W or X, they may form a 12A substituted heterocycle; and wherein R 3A and R 5A One or more of the atoms to which they are attached may form an optionally R 12A Substituted carbocyclic ring, such as heterocyclic ring; L1 is 0, 1, 2, 3, 4, 5 or 6; in: Each R 11A and R 12A are independently selected from 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 ; R 13A and R 14A are each independently selected from hydrogen or R 15A ; R 15A Selected from hydrocarbon groups (such as C 1-6 alkyl, alkenyl, alkynyl or haloalkyl), carbocyclic group and -(CH2) m1 -heterocyclic group, and each R 15A Optionally and independently halogen, cyano, amino, hydroxyl, C 1-6 Alkyl or cycloalkyl and C 1-6 one or more substitutions in the alkoxy group; m1 is 0, 1, 2, 3, 4, 5 or 6; p1 is 0, 1, 2, 3 or 4; R 4A The values ​​of can be the same or different; and q1 is 0, 1, 2, 3 or 4; where R 5A The values ​​of can be the same or different; One or both of Y and Z may be absent and are independently selected from hydrogen, R 16A , optionally by R 16A Substituted hydrocarbon groups (such as C 1-6 alkyl, alkenyl, alkynyl or haloalkyl) and optionally R 16A Substituted -(CH2) r1 A heterocyclic group, wherein each R 16A are independently selected from 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; in: R 17A and R 18A are each independently selected from hydrogen or R 19A ; R 19A Selected from hydrocarbon groups (such as C 1-6 alkyl, alkenyl, alkynyl or haloalkyl), carbocyclic group and -(CH2) s1 -heterocyclic group, and each R 19A Optionally and independently halogen, cyano, amino, hydroxyl, C 1-6 Alkyl and C 1-6 one or more substitutions in the alkoxy group; and s1 is 0, 1, 2, 3, 4, 5, or 6.

10. The ADC or salt thereof according to claim 9, wherein the NMT inhibitor is a compound of formula (IIIa): or a salt thereof, in: n1 is 0 or 1; E 1 is C; W is a (1-4C)alkyl, aryl (e.g. phenyl) or heteroaryl group (e.g. pyridyl); M is selected from C and N; R 3A , R 4A and R 5A are independently selected from hydrogen, R 12A and optionally R 12A substituted (1-3C)alkyl; R 12A are independently selected from 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 ; R 13A and R 14A are each independently selected from hydrogen or (1-4C)hydrocarbyl (eg methyl); Ring D* is an optionally substituted nitrogen-containing 6- or 7-membered heterocyclic ring, wherein each substitutable carbon or nitrogen in Ring D* is optionally and independently replaced by one or more R 7A replace; R 7A independently selected from hydrogen, (1-4C)alkyl, halogen, trifluoromethyl, cyano, thio, nitro or oxo; R 8A is hydrogen; p1 is 0, 1 or 2, where R 4A The values ​​of can be the same or different; q1 is 3, where R 5A The values ​​of can be the same or different; and t1 is 0, 1 or 2, where R 7A The values ​​can be the same or different.

11. The ADC or a salt thereof according to claim 10, wherein the NMT inhibitor is (2,6-dichloro-4-(2-piperazin-1-yl-pyridin-4-yl)-N-(1,3,5-trimethyl-1H-pyrazol-4-yl)-benzenesulfonamide): or a salt thereof.

12. The ADC or a salt thereof according to claim 10, wherein the NMT inhibitor is 2,6-dichloro-N-(5-isobutyl-1,3-dimethyl-1H-pyrazol-4-yl-4-(-4-(2-piperazin-1-yl-pyridin-4-yl)-benzenesulfonamide: 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): or a salt thereof, in: n1 is 1 or 2; n2 is 1 or 2; X 1 Selected by CR x and N; When present, R x is selected from the group consisting of hydrogen, halogen and -C optionally substituted with 1, 2 or 3 substituents. 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; R 1 is selected from the group consisting of: hydrogen; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OCH3 and -OCF3; and -C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from the group consisting of halogen, -CH3, -OCH3 and -OCF3; R 2 is selected from the group consisting of: hydrogen; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OCH3 and -OCF3; and -C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from the group consisting of halogen, -CH3, -OCH3 and -OCF3; or R 1 and R 2 are linked together so that, together with the atoms to which they are attached, they form C 3-6 A cycloalkyl group or a 3- to 6-membered non-aromatic heterocyclic group containing 1 heteroatom selected from the group consisting of O and N, wherein the C 3-6 The cycloalkyl or 3- to 6-membered non-aromatic heterocyclic group is optionally substituted with 1 or 2 substituents, each of which is independently selected from the group consisting of halogen, -OH, -CH3, -OCH3, and -OCF3; R 3 is selected from the group consisting of: hydrogen; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; and -C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent independently selected from the group consisting of halogen, -OH, -CH3, -OCH3 and -OCF3; or R 1 and R 3 are linked together so that together with the atoms to which they are attached they form a 3- to 6-membered non-aromatic heterocyclic group containing 1 N heteroatom, wherein the 3- to 6-membered non-aromatic heterocyclic group is optionally substituted with 1 or 2 substituents, each substituent being independently selected from the group consisting of halogen, -CH3, -OH, -OCH3, and -OCF3; X 2 Selected by CR 4 and N; When present, R 4 is selected from the group consisting of: hydrogen; halogen; and -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from halogen, -OH, -OCH3, -OCF3 and -NR a R b The group composed of; R 5a and R 5d independently selected from the group consisting of: hydrogen; halogen; methyl optionally substituted with 1, 2 or 3 substituents, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; and methoxy optionally substituted with 1, 2 or 3 substituents, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; R 5b and R 5c independently selected from the group consisting of: hydrogen; halogen; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; -OCF3 optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; and C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from the group consisting of halogen, -CH3, -OH, -OCH3, and -OCF3; or R 5b and R 5c connected together so that they, together with the atoms to which they are connected, form a 6-membered aryl group or a 5- or 6-membered aromatic heterocyclic group comprising 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 heterocyclic group is optionally substituted with 1 or 2 substituents, each of which is independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; R 6 selected from the group consisting of hydrogen and methyl; When present, each R 7 is -C optionally substituted by 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; R 8 is selected from the group consisting of: hydrogen; halogen; -OH; -CN; -C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OH, -CN and methoxy optionally substituted by 1, 2 or 3 halogens; -C 3-6 Cycloalkyl, each substituent is independently selected from the group consisting of halogen, -CH3, -OH, -CN and methoxy optionally substituted by 1, 2 or 3 halogens; -C 1-4 alkenyl, each substituent independently selected from the group consisting of halogen, -OH, -CN, and methoxy optionally substituted with 1, 2 or 3 halogens; and -OC optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OH, CN, and methoxy optionally substituted with 1, 2 or 3 halogens; R 9 is selected from hydrogen and -C optionally substituted by 1, 2 or 3 substituents 1-4 alkyl, each substituent being independently selected from the group consisting of halogen, -OH, -OCH3 and -OCF3; or R 8 and R 9 are linked together so that they, together with the atoms to which they are attached, form a 6-membered aromatic group, C 5-6 cycloalkyl, or a 5- or 6-membered aromatic heterocyclic group containing 1 or 2 heteroatoms selected from N, O and S, and wherein the 6-membered aromatic group, C 5-6 The cycloalkyl or 5- to 6-membered aromatic heterocyclic group is optionally substituted by 1, 2 or 3 substituents, each of which is independently selected from the group consisting of: halogen; -OH; -CN; -C optionally substituted by 1, 2 or 3 substituents. 1-4 alkyl, each substituent independently selected from the group consisting of halogen, -OH, and methoxy optionally substituted with 1, 2 or 3 halogens; and -OC optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of -halogen, -OH, and methoxy optionally substituted with 1, 2 or 3 halogens; p is 0, 1, or 2; Z is a 5- to 13-membered non-aromatic heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O and S, wherein at least one heteroatom is N, and wherein the 5- to 13-membered non-aromatic heterocyclic group is optionally substituted with 1, 2, 3 or 4 substituents, each of which is independently selected from the group consisting of: halogen; -C optionally substituted with 1, 2 or 3 substituents 1-6 Alkyl, each substituent is independently selected from halogen, -OH and -OC optionally substituted by 1, 2 or 3 halogens 1-3 alkyl; -OC optionally substituted by 1, 2 or 3 substituents 1-6 Alkyl, each substituent is independently selected from halogen, -OH and -OC optionally substituted by 1, 2 or 3 halogens 1-3 Alkyl group; NR c R d and C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 or when two substituents are in adjacent ring positions, they may be linked together so that together with the atoms to which they are attached they form a C 3-6 Cycloalkyl or a 4- to 6-membered non-aromatic heterocyclic group containing 1 heteroatom selected from O and N, wherein the C 3-6 Cycloalkyl or 4 to 6 membered non-aromatic heterocyclic group is optionally substituted by 1 or 2 substituents, each substituent being independently selected from the group consisting of: halogen; -C 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 alkyl; and -OC optionally substituted by 1, 2 or 3 substituents 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 A group consisting of alkyl groups; R c is hydrogen; R d is selected from the group consisting of: hydrogen; -C optionally substituted with 1, 2 or 3 substituents 1-6 Alkyl, each substituent is independently selected from the group consisting of halogen, -OCH3 and -OCF3; and -C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent being independently selected from the group consisting of halogen, -CH3, -OH, -OCH3, and -OCF3; or Z is -NR 10 R 11 ,in R 10 is hydrogen; and R 11 is a 5- to 10-membered non-aromatic heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O and S, wherein at least one heteroatom is N, and wherein the 5- to 10-membered non-aromatic heterocyclic group is optionally substituted by 1, 2, 3 or 4 substituents independently selected from the group consisting of halogen; -OH; -C optionally substituted by 1, 2 or 3 substituents 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 alkyl; and -OC optionally substituted by 1, 2 or 3 substituents 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 alkyl; and When present, each R a and R b are independently selected from hydrogen and -C 1-4 Alkyl group.

14. The ADC or salt thereof according to claim 13, wherein the NMT inhibitor is a compound of formula (Vq): or a salt thereof, in: R 3a is H or -C optionally substituted by 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OCH3 and -OCF3; R 8a is halogen or C optionally substituted by 1, 2 or 3 substituents 1-4 Alkyl, each substituent independently selected from the group consisting of halogen, -OCH3 and -OCF3; ra is 0, 1, or 2; m a is 1 or 2; R 12a is hydrogen; and When present, each R 13a independently selected from the group consisting of: C optionally substituted with 1, 2 or 3 substituents 1-4 Alkyl, each substituent is independently selected from the group consisting of halogen, -OCH3 and -OCF3; and C optionally substituted by 1, 2 or 3 substituents 3-6 Cycloalkyl, each substituent independently selected from the group consisting of halogen, -CH3, -OCH3 and -OCF3; or When ra is 2 and two R 13a When the groups are located at adjacent ring positions, the two R 13a are linked together so that they, together with the atoms to which they are attached, form a C optionally substituted with 1 or 2 substituents. 3-6 Cycloalkyl, each substituent independently selected from the group consisting of: halogen; -C 1-6 Alkyl, each substituent independently selected from halogen, -OH and -OC optionally substituted with 1, 2 or 3 halogens 1-3 Alkyl group.

15. The ADC or salt thereof according to claim 13 or 14, wherein 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: 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): or a salt thereof, in: R 1 is a group of formula OLA; L is -(CHR 12 ) m -; Each R 12 independently H or C 1-4 alkyl; m is 1, 2 or 3; A is: v is 0, 1, or 2; R 9a H, C 1-4 Alkyl or C 1-4 Haloalkyl; R 9b H, C 1-4 Alkyl or C 1-4 Haloalkyl; R 9c C 1-4 Alkyl or C 1-4 Haloalkyl; R 9d H, C 1-4 Alkyl or C 1-4 Haloalkyl; R 10 H, C 1-4 Alkyl or C 1-4 Haloalkyl; R 11 H, halide, 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 3 halogen groups 1-4 Alkyl, OCH3 or OCF3; Y is CH or C 1-4 alkyl; R 3 H or C 1-4 alkyl; R 4 H or C 1-4 alkyl; R 5 is H; R 6 H or C 1-4 alkyl; q is 0 or 1; R 7 is H or methyl; R 8 is H or methyl; or R 3 With R 6 The atoms between them form bonds between them, or between them and -(CHR a ) r - a 3- to 7-membered non-aromatic heterocyclic ring; or R 7 Group and R 6 The groups and the atoms between them form a a ) r - a 3- to 7-membered non-aromatic heterocyclic ring; r is 1, 2, 3, 4 or 5; and R a It is hydrogen or methyl.

17. The ADC or salt thereof according to claim 16, wherein the NMT inhibitor is a compound of formula (VIA): or a salt thereof, in: R 2a is H or F; R 2b is F; R 5a is H; R 6a is H or methyl; R 9ca is methyl, isopropyl or tert-butyl; R 9cb is H or methyl; R 10a is methyl; and R 11a is methyl; The condition is that when R 2a When H, R 9cb For H.

18. The ADC or salt thereof according to claim 16 or 17, wherein 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.

19. The ADC or salt thereof according to claim 16 or 17, which 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: or a salt thereof.

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

21. The ADC or salt thereof according to claim 20, wherein the linker is cleavable by an enzyme.

22. The ADC or salt thereof according to any one of claims 1 to 21, wherein the linker has the following formula (VII): -A a -W w -Y y -(Formula (VII)) in: A is a first stretcher unit, which, when present, forms a covalent bond with a chain terminus (such as the N-terminus) or a 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, and when A and / or Y are absent, it forms a covalent bond with the chain terminus (e.g., N-terminus) or a functional group of an amino acid side chain of the antibody and / or 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 is a second stretcher unit which, when present, forms a covalent bond with the functional group of the NMT inhibitor; and y is either 0 or 1.

23. The ADC or salt thereof according to claim 22, wherein A has formula (A1): in: n is 1 to 6; represents the point of attachment to the chain terminus (e.g., N-terminus) or a functional group of an amino acid side chain of the antibody; and Indicates the connection point with W.

24. The ADC or salt thereof according to claim 23, wherein A is 25. The ADC or salt thereof according to claim 22, wherein A is 26. The ADC or salt thereof according to any one of claims 22 to 25, wherein each W is an amino acid.

27. The ADC or salt thereof according to claim 26, wherein each W has formula (WI): where w is as defined above; and R 19 is 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, in represents the point of attachment to A (when present) or to a chain terminus (e.g., the N-terminus) or a functional group of an amino acid side chain of the antibody; and represents the point of attachment to Y (when present) or to a functional group of the NMT inhibitor.

28. The ADC or salt thereof according to claim 27, wherein at least one R 19 It is isopropyl.

29. The ADC or salt thereof according to claim 27, wherein at least one R 19 It is (CH2)3NHC(=NH)NH2.

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

31. The ADC or salt thereof according to any one of claims 22 to 30, wherein (W)w has the formula (WII): in: R 19a is H, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, CH2OH, CH(OH)Me, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, phenyl, 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.

32. The ADC or salt thereof according to claim 31, wherein R 19a It is isopropyl.

33. The ADC or salt thereof according to claim 31 or 32, wherein R 19b It is (CH2)3NHC(=NH)NH2.

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

35. The ADC or salt thereof according to claim 34, wherein (W)w has the formula (WIII): in: R is H, halide, CN or NO2; m is 0, 1, 2 or 3; Su is the sugar part; in represents the point of attachment to A (when present) or to a chain terminus (e.g., the N-terminus) or a functional group of an amino acid side chain of the antibody; and represents the point of attachment to Y (when present) or to a functional group of the NMT inhibitor.

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

37. The ADC or salt thereof according to claim 35 or 36, wherein Su has the following formula:

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

39. The ADC or salt thereof according to any one of claims 22 to 33, wherein y is 1 and Y has the formula (Y1): in: Each Q is independently a halide, NO2, CN, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 Haloalkoxy; m is 0 to 4; in represents the point of connection with W; and represents the point of attachment to the functional group of the NMT inhibitor.

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

41. The ADC or salt thereof according to claim 22, wherein the linker has formula (LI): in represents the point of attachment to a functional group on the chain terminus (e.g., N-terminus) or amino acid side chain of the antibody; and represents the point of attachment to the functional group of the NMT inhibitor.

42. The ADC of claim 22, wherein the linker has formula (LII): in represents the point of attachment to a functional group on the chain terminus (e.g., N-terminus) or amino acid side chain of the antibody; and represents the point of attachment to the functional group of the NMT inhibitor.

43. The ADC of claim 20 or 21, wherein the linker has formula (LIII): in represents the point of attachment to a functional group on the chain terminus (e.g., N-terminus) or amino acid side chain of the antibody; and represents the point of attachment to the functional group of the NMT inhibitor.

44. The ADC of claim 20 or 21, wherein the linker has formula (LIV): in represents the point of attachment to a functional group on the chain terminus (e.g., N-terminus) or amino acid side chain of the antibody; and represents the point of attachment to the functional group of the NMT inhibitor.

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

46. ​​The ADC or salt thereof according to any one of claims 22 to 44, wherein the functional group on the NMT inhibitor is an amino group or an alcohol, such as an amino group.

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

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

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

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

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

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

54. The ADC or salt thereof according to claim 53, wherein the antibody is taciturnumab.

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

56. The ADC or salt thereof according to claim 55, wherein the antibody is ifenatumomab.

57. The ADC or salt thereof according to any one of claims 1 to 56, wherein the drug loading (p) of the NMT inhibitor / antibody is 1 to 10 NMT inhibitors / antibody, such as 2 to 6, 4 to 6, 8 to 10 or 6 to 8 NMT inhibitors / antibody.

58. The ADC or salt thereof according to any one of claims 1 to 57, wherein the ADC or salt thereof comprises the formula: or a salt thereof, wherein Ab is an antibody as defined in any one of claims 1 to 56, for example, the antibody is trastuzumab, and p is the drug loading of the NMT inhibitor as defined in claim 57.

59. The ADC or salt thereof according to any one of claims 1 to 57, wherein the ADC or salt thereof comprises the formula: or a salt thereof, wherein Ab is an antibody as defined in any one of claims 1 to 56, for example, the antibody is trastuzumab, and p is the drug loading of the NMT inhibitor as defined in claim 57.

60. The ADC or salt thereof according to any one of claims 1 to 57, wherein the ADC or salt thereof comprises the formula: or a salt thereof, wherein Ab is saccharotuzumab, and p is the drug loading of the NMT inhibitor as defined in claim 57.

61. The ADC or salt thereof according to any one of claims 1 to 57, wherein the ADC or salt thereof comprises the formula: or a salt thereof, wherein Ab is ifenatumomab, and p is the drug loading of the NMT inhibitor as defined in claim 57.

62. The ADC or salt thereof according to any one of claims 1 to 57, wherein the ADC or salt thereof comprises the formula: wherein Ab is an antibody as defined in any one of claims 1 to 56, for example, the antibody is trastuzumab, and p is the drug loading of the NMT inhibitor as defined in claim 57.

63. The ADC or salt thereof according to any one of claims 1 to 57, wherein the ADC or salt thereof comprises the formula: wherein Ab is an antibody as defined in any one of claims 1 to 56, for example, the antibody is trastuzumab, and p is the drug loading of the NMT inhibitor as defined in claim 57.

64. The ADC or salt thereof according to any one of claims 1 to 57, wherein the ADC or salt thereof comprises the formula: wherein Ab is an antibody as defined in any one of claims 1 to 56, for example, the antibody is trastuzumab, and p is the drug loading of the NMT inhibitor as defined in claim 57.

65. The ADC or salt thereof according to any one of claims 1 to 57, wherein the ADC or salt thereof comprises the formula: wherein Ab is an antibody as defined in any one of claims 1 to 56, for example, the antibody is trastuzumab, and p is the drug loading of the NMT inhibitor as defined in claim 57.

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

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

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

69. The ADC according to claim 68, or a pharmaceutically acceptable salt thereof, for use in preventing or treating a disease or condition in which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect.

70. Use of an ADC according to any one of claims 3 to 66, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for preventing or treating a disease or condition in which inhibition of N-myristoyltransferase provides a therapeutic or preventive effect.

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

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

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

74. The ADC for use according to claim 72, or a pharmaceutically acceptable salt thereof, use or method, wherein the cancer is a hematological malignancy (e.g., a lymphoma, in particular 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 lymphoblastic 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. The ADC for use according to claim 72, or a pharmaceutically acceptable salt thereof, use or method, wherein the cancer expresses HER2 protein, such as 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 tumors, 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.

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 functional group, such as a thiol group, on the chain end (e.g., N-terminus) or amino acid side chain of an antibody.

77. The drug conjugate, salt and / or solvate thereof according to claim 76, having the following formula: A a -W w -Y y -NMT (Formula (X)) or a salt and / or solvate thereof, in: A is a first stretching unit, which, when present, comprises a group capable of forming a covalent bond with a functional group, such as a thiol group, on the chain end (such as the N-terminus) or the amino acid side chain of the antibody; NMT is an NMT inhibitor as defined in 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.

78. The drug conjugate, salt and / or solvate thereof according to claim 77, wherein A is in Indicates the connection point with W.

79. The drug conjugate, salt and / or solvate thereof according to any one of claims 76 to 78, wherein the drug conjugate has formula (DC-2): or a salt and / or solvate thereof, wherein NMT is an NMT inhibitor as defined in any one of claims 1 to 19, in particular an NMT inhibitor as defined in any one of claims 16 to 19.

80. The drug conjugate, salt and / or solvate thereof according to any one of claims 76 to 79, which 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-3-yl]methyl}(methyl)carbamoyl)oxy]methyl}-2-[3-(3-{2-[2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy]ethoxy}propionamido)propionamido]phenoxy)-3,4-dihydroxy-2-methylcyclohexane-1-carboxylic acid: or a salt and / or solvate thereof.

81. The drug conjugate, salt and / or solvate thereof according to any one of claims 76 to 79, which is: or a salt and / or solvate thereof.

82. The drug conjugate, salt and / or solvate thereof according to any one of claims 76 to 77, wherein the drug conjugate has formula (DC-1): or a salt and / or solvate thereof, wherein NMT is an NMT inhibitor as defined in any one of claims 1 to 19, in particular an NMT inhibitor as defined in any one of claims 16 to 19.

83. The drug conjugate according to claim 76, 77 or 82, its salt and / or solvate, which is 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-methylcarbamic acid {4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamino]-3-methylbutanoylamino]pentanoylamino]phenyl}methyl ester: or a salt and / or solvate thereof.

84. The drug conjugate, salt and / or solvate thereof according to any one of claims 76 to 79, which is: or a salt and / or solvate thereof.

85. The drug conjugate, salt and / or solvate thereof according to any one of claims 76 to 77, wherein the drug conjugate has formula (DC-5): or a salt and / or solvate thereof, wherein NMT is an NMT inhibitor as defined in any one of claims 1 to 19, in particular an NMT inhibitor as defined in any one of claims 16 to 19.

86. The drug conjugate, salt and / or solvate thereof according to claim 85, which is: or a salt and / or solvate thereof.

87. The drug conjugate, salt and / or solvate thereof according to any one of claims 76 to 77, wherein the drug conjugate has formula (DC-6): or a salt and / or solvate thereof, wherein NMT is an NMT inhibitor as defined in any one of claims 1 to 19, in particular an NMT inhibitor as defined in any one of claims 16 to 19.

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

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

90. A solvate of a salt of a drug conjugate according to any one of claims 76 to 87.

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

92. A compound selected from the group consisting of: -Compound of formula (ADC-I): and -Compound of formula (ADC-II): or a salt and / or solvate thereof, Wherein NMT is an NMT inhibitor as defined in any one of claims 1 to 19, in particular an NMT inhibitor as defined in any one of claims 16 to 19.

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