PNU anthracycline-derived linker-payload, pharmaceutical compositions and uses thereof

Abstract

The present disclosure relates to linker-payloads comprising the structure of Formula (I), and pharmaceutically acceptable salts, solvates, or stereoisomers thereof. The present disclosure also relates to pharmaceutical compositions comprising these compounds, and the use of these compounds, their intermediates, and compositions in the prevention or treatment of cancer and / or tumors. [Formula 1] TIFF2025542248000197.tif76165

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 434,520, filed December 22, 2022, and U.S. Provisional Application No. 63 / 496,986, filed April 19, 2023, the disclosures of which are incorporated herein in their entireties.

[0002] The present disclosure provides linker-payload compounds that contain a PNU anthracycline payload and are useful as components of antibody-drug conjugates. The present disclosure also provides antibody-drug conjugates that contain the linker-payload compounds, compositions thereof, and methods of using them to treat cancer. [Background technology]

[0003] Anthracyclines, a class of drugs used in cancer chemotherapy extracted from the bacterium Streptomyces, are one of the most effective anticancer treatments developed to date, demonstrating efficacy against more cancer types than other classes of chemotherapy agents. These compounds have been used to treat many cancers, including leukemia, lymphoma, breast cancer, gastric cancer, uterine cancer, ovarian cancer, bladder cancer, and lung cancer. However, the usefulness of anthracyclines is limited by their toxicity; therefore, only a small number of anthracyclines or related DNA intercalators are available for clinical use.

[0004] Antibody-drug conjugates represent an innovative therapeutic application that combines the unique high specificity, specificity, and antitumor activity of tumor-specific, but not sufficiently cytotoxic, monoclonal antibodies (mAbs) with the potent cell-killing activity of highly cytotoxic small molecule drugs, such as anthracyclines, which are not suitable for systemic administration alone.

[0005] Antibody-drug conjugates are three-component systems consisting of a cytotoxic payload linked to an antibody via a biodegradable linker. First, the antibody binds to a specific marker (antigen or receptor) on the surface of cancer cells, and then the intact antibody-drug conjugate is internalized into the cancer cells, where the linker is degraded and the payload is released.

[0006] Advances in linker technology are needed to provide improved control over drug pharmacokinetics and improve the delivery and release of cytotoxic payloads in cancer cells. The present disclosure addresses that need. Summary of the Invention

[0007] In one embodiment, the compound of formula (I): [ka] and pharmaceutically acceptable salts thereof, wherein R 2 is H or C1-C6 alkyl, R 1 -XYZR 3 Selected from; R 2 is H or C1-C6 alkyl, or R 1 and R 2 are joined to form a 4- to 6-membered monocyclic heterocycloalkylene group or a 5- to 11-membered bicyclic spirocyclic heterocycloalkylene group, each of which is bonded to a ring carbon atom by -XYZR 3 is replaced by; R 3 teeth, [ka] Selected from; X is -(CH2) n -N(R 4 )-, -R 5 -N(R 4 )-, -(CH2) n -N(R 4 )-C(O)R 5 -, -(CH2) n-N(R 4 )-(C1-C6 alkylene)-N(R 4 )-, 5- or 6-membered monocyclic heterocycloalkylene, -(CH2) n -R 5 -N(R 4 )-, 5-11 membered bicyclic spirocyclic heterocycloalkylene, -(CH2) n -R 5 -, -(CH2) n -R 5 -N(R 4 )C(O)O-(CH2) n R 5 -N(R 4 )-, -(CH2) n -NHC(O)O-(CH2) n -R 5 -N(R 4 )-, -(CH2) n -R 5 -OC(O)-N(R 4 )-(C1-C6 alkylene)-N(R 4 )-, -R 5 -NHC(O)CH2OCH2N(R 4 )C(O)CHN(R 4 )-, -(CH2) n -N(R 4 )C(O)NH-N(R 4 )-, -R 5 -N(R 4 )C(O)CHN(R 4 )-, -(CH2) n -N(R 4 )-(CH2) n -N(R 4 )-, and -R 5 -O-CH2N(R 4 )-selected from; Y is a bond or -C(O)C(R 4 )(R 6 )NH-; Z is -C(O)CH(R 7 )NHC(O)CH(R 8 )NH-; R 4 is H, C 1~6 alkyl, or [ka] and; R 5 is C6~C 10 Arylene, 4-6 membered monocyclic heterocycloalkylene, 5-11 membered bicyclic spirocyclic heterocycloalkylene, C3-C7 monocyclic cycloalkylene, C5-C 11 selected from bridged bicyclic cycloalkylene, -NH-NH-, and -NHC(O)NH-, wherein said 4-6 membered monocyclic heterocycloalkylene may be optionally substituted on a ring carbon atom with an oxo group; R 6 , R 7 and R 8 are each independently selected from H, —CHCHCHNHC(O)NH, benzyl, and a naturally occurring amino acid side chain; R 9 is -CH2NHC(O)-(CH2CH2O) q -CH3; m is 0, 1 or 2; each occurrence of n is independently an integer from 0 to 4; q is selected from 4, 8, 10, and 12 is provided.

[0008] The compound of formula (I) and its pharmaceutically acceptable salts can be useful as components of antibody-drug conjugates useful for the treatment and prevention of cancer. Without being bound by any particular theory, it is believed that the payload portion of the compound of formula (I) acts as an inhibitor of topoisomerase.

[0009] Accordingly, provided herein is an antibody-drug conjugate comprising a compound of formula (I) as a payload / linker moiety. Also provided is a method for treating or preventing cancer in a patient, comprising administering to the patient an effective amount of at least one antibody-drug conjugate comprising a compound of formula (I).

[0010] Further details are provided below in the accompanying detailed description.

[0011] Although any methods and materials similar to those described herein can be used in the practice or testing of compounds of Formula (I) or antibody drug conjugates comprising compounds of Formula (I), illustrative methods and materials are now described. Other embodiments, aspects and features are further described in or will be apparent from the ensuing description, examples and appended claims. DETAILED DESCRIPTION OF THE INVENTION

[0012] The present disclosure relates to a class of PNU anthracycline-derived linker-payload compounds ("linker-payload compounds of the present disclosure") in which the linker structure comprises a maleimide group or a sulfone-substituted pyridyl group attached to a peptide linker, which varies in amino acid sequence and may incorporate PEG units, terminating in a connection to a PNU anthracycline-derived payload. Certain embodiments of the present disclosure relate to linker-payload compounds. Another embodiment of the present disclosure relates to antibody-drug conjugates ("ADCs of the present disclosure") comprising linker-payload compounds of the present disclosure. Yet other embodiments relate to novel linker moieties of linker-payload compounds.

[0013] Terms used herein have their usual meanings, and the meaning of such terms is independent at each occurrence. Nevertheless, unless otherwise specified, the following definitions apply throughout this specification and claims. Chemical names, common names, and chemical structures may be used interchangeably to describe the same structure. When a chemical compound is referred to using both a chemical structure and a chemical name, it should be understood that if there is ambiguity between the structure and the name, the structure shall prevail. Unless otherwise specified, these definitions apply regardless of whether the term is used alone or in combination with other terms. Thus, the definition of "alkyl" applies to "alkyl" as well as the "alkyl" portion of "hydroxyalkyl," "haloalkyl," "-O-alkyl," etc.

[0014] As used herein, and throughout this disclosure, the following terms, unless otherwise indicated, shall be understood to have the meanings indicated below.

[0015] Terms used herein have their usual meanings, and the meaning of such terms is independent at each occurrence. Nevertheless, unless otherwise specified, the following definitions apply throughout this specification and claims. Chemical names, common names, and chemical structures may be used interchangeably to describe the same structure. When a chemical compound is referred to using both a chemical structure and a chemical name, it should be understood that if there is ambiguity between the structure and the name, the structure shall prevail. Unless otherwise specified, these definitions apply regardless of whether the term is used alone or in combination with other terms. Thus, the definition of "alkyl" applies to "alkyl" as well as the "alkyl" portion of "hydroxyalkyl," "haloalkyl," "-O-alkyl," etc.

[0016] As used herein, and throughout this disclosure, the following terms, unless otherwise indicated, shall be understood to have the following meanings:

[0017] The term "anticancer agent" refers to chemical compounds that can be used to treat cancer. This definition also includes antihormonal agents, which act to regulate, reduce, block, or inhibit the effects of hormones that promote cancer growth, often in the form of systemic or holistic therapy. Anticancer agents can be hormones.

[0018] The term "compound(s) of the present disclosure" refers to the chemical compounds disclosed herein. Included within the definition of "compound(s) of the present disclosure" are linker-payload compounds of formulas (I) through (XXII), the linker-payload compounds of Examples 1 through 36, and linker-payload compounds comprising (i) a linker selected from the linkers numbered L-1 through L-12 and (ii) a payload of formula (XVI), all of which are collectively referred to herein as "linker-payload compounds of the present disclosure." Also included within the definition of "compound(s) of the present disclosure" are antibody-drug conjugates disclosed herein, including, but not limited to, compounds of formulas (XXIII) through (LVII), and the antibody-drug conjugates of Examples 37 through 72, collectively referred to herein as "ADCs of the present disclosure."

[0019] A "patient" is a human or non-human mammal. In one embodiment, the patient is a human.

[0020] As used herein, the term "effective amount" refers to an amount of a subject compound and / or additional therapeutic agent, or composition thereof, effective upon administration to produce the desired therapeutic, ameliorative, inhibitory, or preventative effect. For the combination therapies described herein, the effective amount can refer to each individual agent or the combination as a whole; the amounts of all agents administered are effective together, but the component agents of the combination may not be present in individually effective amounts.

[0021] The term "preventing," as used herein with respect to a cell proliferative disorder, refers to reducing the likelihood of the cell proliferative disorder.

[0022] As used herein, the terms "treating" or "treatment" (e.g., of a disease, disorder, or condition or associated symptoms, which may together or individually be referred to as an "indication") include inhibiting the disease, disorder, or condition, i.e., halting or reducing the development of the disease or its biological process or its progression or clinical symptoms; or alleviating the disease, i.e., causing regression of the disease or its biological process or its progression and / or clinical symptoms. As used herein, "treatment" also refers to controlling, ameliorating, or reducing the risk to a subject suffering from a disease, disorder, or condition involving a tumor. As used herein, the terms "preventing" or "prevention" or "prophylaxis" of a disease, disorder, or condition include, for example, impeding the onset or progression of clinical symptoms of the disease, disorder, or condition in a mammal that may be exposed to or predisposed to the disease, disorder, or condition, but that has not yet experienced or exhibited symptoms of the disease.

[0023] As used herein, the term "DAR" or "drug-antibody ratio" refers to the average number of linker / drug moieties attached to an antibody in a composition comprising multiple ADCs of the present disclosure. For a composition comprising an ADC of the present disclosure, the DAR of the composition is the average of the DARs of all individual antibody-drug conjugate molecules present in the composition, with this average expressed as a decimal. Thus, in some embodiments of a composition comprising an ADC of the present disclosure, the DAR of the composition is a decimal number between 0 and 8, 0 and 7, 0 and 6, 0 and 5, 0 and 4, 0 and 3, 0 and 2, and 0 and 1. In additional embodiments, for a composition comprising an ADC of the present disclosure, the DAR of the composition is a decimal number between 1 and 4, 2 and 5, 3 and 6, 4 and 7, 5 and 8, and 6 and 8. In other embodiments, for a composition comprising an ADC of the present disclosure, the DAR of the composition is a decimal number between 1 and 3, 2 and 4, 3 and 5, 4 and 6, 5 and 7, and 6 and 8. In further embodiments, for compositions comprising an ADC of the disclosure, the DAR of the composition is a decimal number between 1 and 2, between 2 and 3, between 3 and 4, between 4 and 5, between 5 and 6, between 6 and 7, and between 7 and 8. The term "composition" as used above is understood to encompass pharmaceutical compositions.

[0024] As used herein, the term "alkyl" refers to an aliphatic hydrocarbon group having one of its hydrogen atoms replaced with a bond. An alkyl group can be straight or branched and can contain from about 1 to about 20 carbon atoms. In one embodiment, an alkyl group contains from about 1 to about 10 carbon atoms. In a different embodiment, an alkyl group can contain from 1 to 10 carbon atoms ("C1-C 10

[0023] Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, neopentyl, isopentyl, n-hexyl, isohexyl, and neohexyl. Alkyl groups can be unsubstituted or substituted with one or more substituents, which can be the same or different, each independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH, -NH(alkyl), -N(alkyl), NH(cycloalkyl), -OC(O)-alkyl, -OC(O)-aryl, -OC(O)-cycloalkyl, -C(O)OH, and -C(O)O-alkyl. In one embodiment, an alkyl group is linear. In another embodiment, an alkyl group is branched. Unless otherwise specified, an alkyl group is unsubstituted.

[0025] As used herein, the term "alkenyl" refers to an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and having one of its hydrogen atoms replaced with a bond. Alkenyl groups can be straight-chained or branched and can contain from about 2 to about 15 carbon atoms. In one embodiment, an alkenyl group contains from about 2 to about 10 carbon atoms. In another embodiment, an alkenyl group contains from about 2 to about 6 carbon atoms. Non-limiting examples of alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, octenyl, and decenyl. An alkenyl group can be unsubstituted or substituted with one or more substituents, which may be the same or different, each substituent independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH, -NH(alkyl), -N(alkyl), -NH(cycloalkyl), -OC(O)-alkyl, -OC(O)-aryl, -OC(O)-cycloalkyl, -C(O)OH and -C(O)O-alkyl. 10 The term "alkenyl" refers to an alkenyl group having from 2 to 10 carbon atoms. Unless otherwise specified, the alkenyl group is unsubstituted.

[0026] The term "alkynyl," as used herein, refers to an aliphatic hydrocarbon group containing at least one carbon-carbon triple bond and having one of its hydrogen atoms replaced with a bond. Alkynyl groups can be straight-chained or branched and can contain from about 2 to about 15 carbon atoms. In one embodiment, an alkynyl group contains from about 2 to about 10 carbon atoms. In another embodiment, an alkynyl group contains from about 2 to about 6 carbon atoms. Non-limiting examples of alkynyl groups include ethynyl, propynyl, 2-butynyl, and 3-methylbutynyl. An alkynyl group can be unsubstituted or substituted with one or more substituents, which may be the same or different, each substituent independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH, -NH(alkyl), -N(alkyl), -NH(cycloalkyl), -OC(O)-alkyl, -OC(O)-aryl, -OC(O)-cycloalkyl, -C(O)OH and -C(O)O-alkyl. 10 The term "alkynyl" refers to an alkynyl group having from 2 to 10 carbon atoms. Unless otherwise specified, the alkynyl group is unsubstituted.

[0027] The term "alkylene," as used herein, refers to an alkyl group, as defined above, where one of the alkyl group's hydrogen atoms has been replaced with a bond. Non-limiting examples of alkylene groups include -CH-, -CHCH-, -CHCHCH-, -CHCHCHCH-, -CH(CH)CHCH-, -CH(CH)-, and -CHCH(CH)CH-. In one embodiment, an alkylene group has 1 to about 10 carbon atoms. In another embodiment, an alkylene group has 1 to about 6 carbon atoms. In another embodiment, an alkylene group is branched. In another embodiment, an alkylene group is linear. In one embodiment, an alkylene group is -CH-. The term "C1-C6 alkylene" refers to an alkylene group having 1 to 6 carbon atoms.

[0028] The term "alkenylene," as used herein, refers to an alkenyl group, as defined above, in which one of the alkenyl group's hydrogen atoms has been replaced with a bond. Non-limiting examples of alkylene groups include -CH=CH-, -CH=CHCH2-, -CH2CH2CH=CH-, and -CH2(CH3)C=CH-. In one embodiment, an alkenylene group has 2 to about 6 carbon atoms. In one embodiment, an alkenylene group has 2 to about 10 carbon atoms. In another embodiment, an alkenylene group is branched. In another embodiment, an alkenylene group is linear. The term "C2-C6 alkenylene" refers to an alkenylene group having 2 to 6 carbon atoms.

[0029] The term "alkynylene," as used herein, refers to an alkynyl group, as defined above, where one of the alkynyl group's hydrogen atoms has been replaced with a bond. Non-limiting examples of alkylene groups include -C≡C-, -C≡CCH2-, and -C≡CCH(CH3)2-. In one embodiment, an alkynylene group has 2 to about 6 carbon atoms. In another embodiment, an alkynylene group has 2 to about 10 carbon atoms. In another embodiment, an alkynylene group is branched. In another embodiment, an alkynylene group is linear. The term "C2-C6 alkynylene" refers to an alkynylene group having 2 to 6 carbon atoms. "C2-C 10 The term "alkynylene" refers to an alkynylene group having 2 to 10 carbon atoms.

[0030] The term "aminoalkyl," as used herein, refers to an alkyl group, as defined above, in which one of the alkyl group's hydrogen atoms has been replaced with -NH, -NH(C-C alkyl), or -N(C-C alkyl). In one embodiment, the aminoalkyl group has 1 to 6 carbon atoms. Non-limiting examples of aminoalkyl groups include -CHNH, -CHN(CH), -CHCHNH, and -CHNH(CH). The term "C-C aminoalkyl" refers to an aminoalkyl group having 1 to 6 carbon atoms.

[0031] As used herein, the term "antibody" is used in the broadest sense and specifically encompasses intact monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments that exhibit the desired biological activity, so long as the antibody fragment has the required number of binding sites for the drug-linker. The native form of an antibody is a tetramer and consists of two identical immunoglobulin chain pairs, each pair having one light chain and one heavy chain. In each pair, a light chain variable region and a heavy chain variable region (V L and V H ) are primarily responsible for binding to antigens. The light chain variable domain and the heavy chain variable domain consist of a framework region interrupted by three hypervariable regions, also called "complementarity-determining regions" or "CDRs." The constant regions are recognized by and can interact with the immune system (see, e.g., Janeway et al., 2001, Immuno. Biology, 5th Ed., Garland Publishing, New York). Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. Antibodies can be derived from any suitable species. In some embodiments, the antibodies are of human or murine origin. Antibodies can be, for example, human, humanized, or chimeric.

[0032] As used herein, the term "aryl" refers to an aromatic monocyclic or polycyclic ring system containing from about 6 to about 14 carbons. In one embodiment, an aryl group contains from about 6 to about 10 carbon atoms ("C6-C 10 Aryl). The aryl group can be optionally substituted with one or more "ring system substituents," which may be the same or different, and are defined herein below. In one embodiment, the aryl group can be optionally fused to a cycloalkyl or cycloalkanoyl group. Non-limiting examples of aryl groups include phenyl and naphthyl. Examples of aryl groups fused to a cycloalkyl ring include: [ka] Examples include:

[0033] In one embodiment, the aryl group is phenyl. In another embodiment, the aryl group is naphthalene. Unless otherwise specified, alkyl groups are unsubstituted.

[0034] The term "arylene," as used herein, refers to an aryl group, as defined above, in which one of the aryl group's hydrogen atoms has been replaced with a bond. Non-limiting examples of arylene groups include: [ka] Examples include:

[0035] In one embodiment, an alkylene group has from 1 to about 10 carbon atoms. In another embodiment, an alkylene group has from 1 to about 6 carbon atoms. In another embodiment, an alkylene group is branched. In another embodiment, an alkylene group is linear. In one embodiment, an alkylene group is -CH2-. The term "C1-C6 alkylene" refers to an alkylene group having from 1 to 6 carbon atoms.

[0036] As used herein, the term "composition" is intended to encompass a product comprising an ADC of the present disclosure, or a pharmaceutically acceptable salt thereof, together with one or more additional specified ingredients in a specified amount, as well as any product resulting directly or indirectly from the combination of the specified ingredients in the specified amounts. This term, with respect to pharmaceutical compositions, is intended to encompass products comprising an ADC of the present disclosure, or a pharmaceutically acceptable salt thereof, optionally together with one or more additional active ingredients, active ingredient(s), and inert ingredient(s) comprising the carrier, as well as any product resulting directly or indirectly from the combination, complexation, or aggregation of any two or more of the ingredients, or from the dissociation of one or more of the ingredients, or from one or more other types of reaction or interaction of the ingredients. Thus, pharmaceutical compositions of the present disclosure encompass any composition prepared by admixing an ADC of the present disclosure, or a pharmaceutically acceptable salt thereof, with a pharmaceutically acceptable carrier. "Pharmaceutically acceptable" means that the carrier, diluent, or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0037] The term "cycloalkyl," as used herein, refers to a non-aromatic monocyclic or polycyclic ring system containing about 3 to about 11 ring carbon atoms. In one embodiment, a cycloalkyl contains about 5 to about 11 ring carbon atoms. In another embodiment, a cycloalkyl is monocyclic and contains about 3 to about 7 ring atoms. In another embodiment, a cycloalkyl is monocyclic and contains about 5 to about 6 ring atoms. In another embodiment, a cycloalkyl is bicyclic and contains about 4 to 10 ring atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Non-limiting examples of polycyclic cycloalkyls include 1-decalinyl, norbornyl, and adamantyl. A cycloalkyl group can be optionally substituted with one or more "ring system substituents," which may be the same or different, and are defined herein below. Unless otherwise specified, a cycloalkyl group is unsubstituted. In one embodiment, a cycloalkyl group is unsubstituted. The term "3- to 7-membered monocyclic cycloalkyl" refers to a monocyclic cycloalkyl group having from 3 to 7 ring carbon atoms. The term "5- to 11-membered bicyclic cycloalkyl group" refers to a bicyclic cycloalkyl group having from 5 to 11 ring carbon atoms.

[0038] Polycyclic cycloalkyl groups can have fused rings, rings connected in a spirocyclic fashion, and bridged rings. In one embodiment, the cycloalkyl group is a spirocyclic cycloalkyl group having from 5 to 11 ring carbon atoms ("C5-C 11 Illustrative examples of such bicyclic cycloalkyl groups include: [ka] Examples include:

[0039] In another embodiment, the cycloalkyl group is a fused bicyclic cycloalkyl group having 5 to 11 ring carbon atoms ("C5-C 11Illustrative examples of such fused bicyclic cycloalkyl groups include: [ka] Examples include:

[0040] In another embodiment, the cycloalkyl group is a bridged bicyclic cycloalkyl group having 5 to 11 ring carbon atoms ("C5-C 11 The group may be a bridged bicyclic cycloalkyl group ("bridged bicyclic cycloalkyl"), or a bridged tricyclic cycloalkyl group having 6 to 14 ring carbon atoms. Illustrative examples of such bridged bicyclic and tricyclic heterocycloalkyl groups include: [ka] Examples include:

[0041] A ring carbon atom of a cycloalkyl group may be functionalized as a carbonyl group. Illustrative examples of such cycloalkyl groups (also referred to herein as "cycloalkanoyl" groups) include, but are not limited to, cyclobutanoyl: [ka] Examples include:

[0042] The term "cycloalkylene," as used herein, refers to a cycloalkyl group, as defined above, in which one of the cycloalkyl group's hydrogen atoms has been replaced with a bond. In one embodiment, the cycloalkylene is monocyclic and contains from about 3 to about 7 ring carbon atoms ("C-C monocyclic cycloalkylene"). In another embodiment, the cycloalkylene is bicyclic and contains from about 5 to 10 ring atoms ("C-C 10 Non-limiting examples of monocyclic cycloalkylenes include: [ka] Examples include:

[0043] A cycloalkylene group can be optionally substituted with one or more "ring system substituents," which may be the same or different, and are defined herein below. Unless otherwise specified, a cycloalkylene group is unsubstituted. In one embodiment, a cycloalkylene group is unsubstituted.

[0044] Polycyclic cycloalkylene groups can have fused rings, rings connected in a spirocyclic fashion, and bridged rings. In one embodiment, the cycloalkylene group can be a bridged bicyclic cycloalkylene group having 5 to 11 ring carbon atoms. Illustrative examples of such bridged bicyclic heterocycloalkyl groups include, but are not limited to: [ka] Examples include:

[0045] A ring carbon atom of a cycloalkylene group may be functionalized as a carbonyl group. Illustrative examples of such cycloalkylene groups include, but are not limited to: [ka] Examples include:

[0046] The term "cycloalkenyl," as used herein, refers to a non-aromatic monocyclic or polycyclic ring system containing about 4 to about 10 ring carbon atoms and containing at least one endocyclic double bond. In one embodiment, a cycloalkenyl contains about 4 to about 7 ring carbon atoms. In another embodiment, a cycloalkenyl contains 5 or 6 ring atoms. Non-limiting examples of monocyclic cycloalkenyls include cyclopentenyl, cyclohexenyl, cyclohepta-1,3-dienyl, and the like. A cycloalkenyl group can be optionally substituted with one or more "ring system substituents," which may be the same or different, and are defined herein below. A ring carbon atom of a cycloalkylene group can be functionalized as a carbonyl group. In one embodiment, a cycloalkenyl group is cyclopentenyl. In another embodiment, a cycloalkenyl group is cyclohexenyl. The term "4- to 6-membered cycloalkenyl" refers to a cycloalkenyl group having 4 to 6 ring carbon atoms.

[0047] The term "halo" as used herein means -F, -Cl, -Br or -I.

[0048] The term "haloalkyl," as used herein, refers to an alkyl group, as defined above, in which one or more of the alkyl group's hydrogen atoms has been replaced with a halogen. In one embodiment, a haloalkyl group has 1 to 10 carbon atoms. In another embodiment, a haloalkyl group has 1 to 6 carbon atoms. In another embodiment, a haloalkyl group is substituted with 1 to 6 F atoms. Within a class of this embodiment, a haloalkyl group is substituted with 1 to 3 F atoms. Non-limiting examples of haloalkyl groups include -CH2CHF2, -CH2F, -CHF2, -CF3, -CH2Cl, and -CCl3. The term "C1-C6 haloalkyl" refers to a haloalkyl group having 1 to 6 carbon atoms.

[0049] The term "haloalkylene," as used herein, refers to a haloalkyl group, as defined above, in which one or more of the haloalkyl group's hydrogen atoms has been replaced with a halogen. In one embodiment, a haloalkylene group has 1 to 10 carbon atoms. In another embodiment, a haloalkylene group has 1 to 6 carbon atoms. In another embodiment, a haloalkylene group is substituted with 1 to 6 F atoms. Within a class of this embodiment, a haloalkylene group is substituted with 1 to 3 F atoms. Non-limiting examples of haloalkylene groups include -CHCHF, -CHF, -CHF, -CF, -CHCl, and -CCl. The term "C-C haloalkylene" refers to a haloalkylene group having 1 to 6 carbon atoms.

[0050] The term "hydroxyalkyl," as used herein, refers to an alkyl group, as defined above, where one or more of the alkyl group's hydrogen atoms have been replaced with an -OH group. In one embodiment, the hydroxyalkyl group has 1 to 10 carbon atoms. In another embodiment, the hydroxyalkyl group has 1 to 6 carbon atoms. Non-limiting examples of hydroxyalkyl groups include -CHOH, -CHCHOH, -CHCHCHOH, and -CHCH(OH)CH. "C1-C 10 The term "hydroxyalkyl" refers to a hydroxyalkyl group having from 1 to 10 carbon atoms.

[0051] The term "heteroaryl," as used herein, refers to an aromatic monocyclic or polycyclic ring system containing about 5 to about 14 ring atoms, wherein 1 to 4 of the ring atoms are independently O, N, or S, and the remaining ring atoms are carbon atoms. In one embodiment, a heteroaryl group has 5 to 10 ring atoms. In another embodiment, a heteroaryl group is monocyclic and has 5 or 6 ring atoms (a "5- or 6-membered monocyclic heteroaryl"). In another embodiment, a heteroaryl group is bicyclic and has 8 to 10 ring atoms (an "8- to 10-membered bicyclic heteroaryl"). In yet another embodiment, a heteroaryl group is bicyclic and has 9 or 10 ring atoms (a "9- or 10-membered bicyclic heteroaryl"). A heteroaryl group can be optionally substituted with one or more "ring system substituents," which may be the same or different, and are defined herein below. Heteroaryl groups are joined via ring carbon atoms, and any nitrogen atom of a heteroaryl may be optionally oxidized to the corresponding N-oxide. The term "heteroaryl" also encompasses heteroaryl groups as defined above fused to a benzene ring. Non-limiting examples of heteroaryls include pyridyl, pyrazinyl, furanyl, thienyl, pyrimidinyl, pyridone (including N-substituted pyridones), isoxazolyl, isothiazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyrazolyl, furazanyl, pyrrolyl, triazolyl, 1,2,4-thiadiazolyl, pyrazinyl, pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl, imidazo[1,2-a]pi Examples of heteroaryl include lysinyl, imidazo[2,1-b]thiazolyl, benzofurazanyl, indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl, imidazolyl, benzimidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl, pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl, 1,2,4-triazinyl, benzothiazolyl, and the like, and all isomeric forms thereof. The term "heteroaryl" also refers to partially saturated heteroaryl moieties, such as tetrahydroisoquinolyl, tetrahydroquinolyl, and the like. In one embodiment, the heteroaryl group is a 5-membered heteroaryl.In another embodiment, the heteroaryl group is a 6-membered heteroaryl, for example, pyridyl.

[0052] In one embodiment, the 8-10 membered bicyclic heteroaryl group is a fused bicyclic heterocyclic group in which one of the two fused rings is phenyl or a monocyclic heteroaryl, such as [ka] Includes.

[0053] "9-14 membered tricyclic heteroaryl" includes 8-10 membered bicyclic heteroaryl groups in which the third ring is fused to one of the rings of the 8-10 membered bicyclic heteroaryl group. Such a third ring can be a cycloalkyl, heterocycloalkyl, or heteroaryl ring. Examples of 9-14 membered tricyclic heteroaryl groups include: [ka] [ka] The term "heterocycloalkyl," as used herein, refers to a non-aromatic saturated monocyclic or polycyclic ring system containing 3 to about 14 ring atoms, wherein 1 to 4 of the ring atoms are independently O, S, N, or Si, and the remainder of the ring atoms are carbon atoms. A heterocycloalkyl group can be joined through a ring carbon, ring silicon, or ring nitrogen atom. In one embodiment, a heterocycloalkyl group is monocyclic. In one embodiment, a heterocycloalkyl group is monocyclic and has about 3 to about 7 ring atoms (a "3- to 7-membered monocyclic heterocycloalkyl"). In another embodiment, a heterocycloalkyl group is monocyclic and has 5 or 6 ring atoms (a "5- or 6-membered monocyclic heterocycloalkyl"). In one embodiment, a heterocycloalkyl group is bicyclic. In another embodiment, a heterocycloalkyl group is bicyclic and has about 5 to about 11 ring atoms (a "5- to 11-membered bicyclic heterocycloalkyl"). In another embodiment, a heterocycloalkyl group is tricyclic and has about 10 to about 14 ring atoms ("10-14-membered tricyclic heterocycloalkyl"). There are no adjacent oxygen and / or sulfur atoms present in the ring system. Any -NH groups in a heterocycloalkyl ring can be present protected, for example, as an -N(BOC), -N(CBz), -N(Tos) group, etc., and such protected heterocycloalkyl groups are considered part of the present disclosure. A heterocycloalkyl group can be optionally substituted with one or more "ring system substituents," which may be the same or different, and are defined herein below. The nitrogen or sulfur atom of a heterocycloalkyl can be optionally oxidized to the corresponding N-oxide, S-oxide, or S,S-dioxide. Non-limiting examples of monocyclic heterocycloalkyl rings include oxetanyl, piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, delta-lactam, delta-lactone, silacyclopentane, silapyrrolidine, and the like, and all isomeric forms thereof. Non-limiting examples of silyl-containing heterocycloalkyl groups include: [ka] Examples include:

[0054] A ring carbon atom of a heterocycloalkyl group may be functionalized as a carbonyl group. Illustrative examples of such heterocycloalkyl groups include, but are not limited to: [ka] Examples include:

[0055] The ring sulfur atom of a heterocycloalkyl group may also be functionalized as a sulfonyl group. Examples of such heterocycloalkyl groups are: [ka] is.

[0056] In one embodiment, a heterocycloalkyl group is a 5-membered monocyclic heterocycloalkyl. In another embodiment, a heterocycloalkyl group is a 6-membered monocyclic heterocycloalkyl.

[0057] Polycyclic heterocycloalkyl groups can have fused rings, rings connected in a spirocyclic fashion, and bridged rings. In one embodiment, a heterocycloalkyl group can be a bicyclic spirocyclic heteroaryl group having 1 to 11 ring atoms. Illustrative examples of such bicyclic heterocycloalkyl groups include: [ka] Examples include:

[0058] In another embodiment, a heterocycloalkyl group can be a fused bicyclic heterocycloalkyl group having from 5 to 11 ring atoms (a "5- to 11-membered fused bicyclic heterocycloalkyl"). Illustrative examples of such fused bicyclic heterocycloalkyl groups include: [ka] Examples include:

[0059] In another embodiment, the heterocycloalkyl group can be a bridged heterocycloalkyl group having 5 to 11 ring atoms (a "5- to 11-membered bridged bicyclic heterocycloalkyl"). Illustrative examples of such bridged bicyclic heterocycloalkyl groups include: [ka] Examples include:

[0060] The term "heterocycloalkylene," as used herein, refers to a heterocycloalkyl group, as defined above, in which one of the heterocycloalkyl group's hydrogen atoms has been replaced with a bond. A heterocycloalkylene group can be linked via a ring carbon atom or a ring nitrogen atom. In one embodiment, a heterocycloalkylene group has 4 to 6 ring atoms (a "4- to 6-membered heterocycloalkylene"). In another embodiment, a heterocycloalkenyl group is monocyclic and has 5 or 6 ring atoms. In another embodiment, a heterocycloalkenyl group is bicyclic and has 5 to 10 ring atoms (a "5- to 10-membered bicyclic heterocycloalkylene"). A heterocycloalkylene group can be optionally substituted with one or more ring system substituents (where "ring system substituent" is defined above). The nitrogen or sulfur atom of a heterocycloalkylene can be optionally oxidized to the corresponding N-oxide, S-oxide, or S,S-dioxide. A ring carbon atom of a heterocycloalkylene group may be functionalized as a carbonyl group. Non-limiting examples of monocyclic heterocycloalkylene groups include: [ka] Examples include:

[0061] Polycyclic heterocycloalkylene groups can have fused rings, rings linked in a spirocyclic fashion, and bridged rings. In one embodiment, a heterocycloalkylene group can be a bicyclic spirocyclic heterocycloalkylene group having 5 to 11 ring atoms ("5-11 membered bicyclic spirocyclic heterocycloalkylene"). Illustrative examples of such bicyclic spirocyclic heterocycloalkylene groups include: [ka] Examples include:

[0062] In another embodiment, the heterocycloalkylene group can be a fused bicyclic heterocycloalkylene group having from 5 to 11 ring atoms (a "5- to 11-membered fused bicyclic heterocycloalkylene"). Illustrative examples of such fused bicyclic heterocycloalkylene groups include: [ka] Examples include:

[0063] In another embodiment, the heterocycloalkylene group can be a bridged heterocycloalkylene group having 5 to 11 ring atoms (a "5- to 11-membered bridged bicyclic heterocycloalkylene"). Illustrative examples of such bridged bicyclic heterocycloalkylene groups include: [ka] Examples include:

[0064] The term "heterocycloalkenyl," as used herein, refers to a heterocycloalkyl group, as defined above, containing 4 to 10 ring atoms and at least one endocyclic carbon-carbon or carbon-nitrogen double bond. A heterocycloalkenyl group can be linked through a ring carbon or a ring nitrogen atom. In one embodiment, a heterocycloalkenyl group has 4 to 6 ring atoms. In another embodiment, a heterocycloalkenyl group is monocyclic and has 5 or 6 ring atoms. In another embodiment, a heterocycloalkenyl group is bicyclic. A heterocycloalkenyl group can be optionally substituted with one or more ring system substituents ("ring system substituent" is defined above). The nitrogen or sulfur atom of a heterocycloalkenyl can be optionally oxidized to the corresponding N-oxide, S-oxide, or S,S-dioxide. A ring carbon atom of a heterocycloalkenyl group can be functionalized as a carbonyl group. Non-limiting examples of heterocycloalkenyl groups include 1,2,3,4-tetrahydropyridinyl, 1,2-dihydropyridinyl, 1,4-dihydropyridinyl, 1,2,3,6-tetrahydropyridinyl, 1,4,5,6-tetrahydropyrimidinyl, 2-pyrrolinyl, 3-pyrrolinyl, 2-imidazolinyl, 2-pyrazolinyl, dihydroimidazolyl, dihydrooxazolyl, dihydrooxadiazolyl, dihydrothiazolyl, 3,4-dihydro-2H-pyranyl, dihydrofuranyl, fluoro-substituted dihydrofuranyl, 7-oxabicyclo[2.2.1]heptenyl, dihydrothiophenyl, dihydrothiopyranyl, and the like. In one embodiment, a heterocycloalkenyl group is a 5-membered heterocycloalkenyl. In another embodiment, a heterocycloalkenyl group is a 6-membered heterocycloalkenyl. The term "4 to 6-membered heterocycloalkenyl" refers to a heterocycloalkenyl group having 4 to 6 ring atoms.

[0065] The term "substituted" means that one or more hydrogens on the designated atom have been replaced with a selection from the indicated group, provided that the replacement does not exceed the normal valence of the designated atom under the conditions present, and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds. By "stable compound" or "stable structure" is meant a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

[0066] As used herein, the term "in substantially purified form" refers to the physical state of a compound after it has been isolated from a synthetic method (e.g., from a reaction mixture), a natural source, or a combination thereof. The term "in substantially purified form" also refers to the physical state of a compound after it has been obtained from one or more purification methods described herein or known to those of skill in the art (e.g., chromatography, recrystallization, etc.) of sufficient purity that it can be characterized by standard analytical techniques described herein or known to those of skill in the art.

[0067] It should also be noted that any carbon and heteroatom with unsatisfied valences in the text, schemes, examples and tables herein is assumed to have sufficient hydrogen atoms to satisfy the valences.

[0068] When a functional group of a compound is referred to as "protected," this means that the group is modified to prevent undesired side reactions at the protected site when the compound undergoes a reaction. Suitable protecting groups will be recognized by those skilled in the art and by reference to standard texts such as, for example, Greene et al., Protective Groups in Organic Synthesis, Wiley-Interscience, New York, (1999).

[0069] Examples of "ring system substituents" include, but are not limited to, alkyl, alkenyl, alkynyl, aryl, heteroaryl, -alkylene-aryl, -arylene-alkyl, -Alkylene-heteroaryl, -alkenylene-heteroaryl, -alkynylene-heteroaryl, -OH, hydroxyalkyl, haloalkyl, -O-alkyl, -O-haloalkyl, -alkylene-O-alkyl, -O-aryl, -O-alkylene-aryl, acyl, -C(O)-aryl, halo, -NO2, -CN, -SF5, -C(O)OH, -C(O)O-alkyl, -C(O)O-aryl, -C(O)O-alkylene-aryl, -S(O)-alkyl, -S(O)2-alkyl, -S(O)-aryl, -S(O)2-aryl, -S(O)-heteroaryl, -S(O)z-heteroaryl, -S-aryl alkyl, -S-aryl, -S-heteroaryl, -S-alkylene-aryl, -S-alkyleneheteroaryl, -S(O)2-alkylene-aryl, -S(O)2-alkylene-heteroaryl, -Si(alkyl)2, -Si(aryl)2, Si(heteroaryl)2, -Si(alkyl)(aryl), -Si(alkyl)(cycloalkyl), -Si(alkyl)(heteroaryl), cycloalkyl, heterocycloalkyl, -OC(O)-alkyl, -OC(O)-aryl, -OC(O)-cycloalkyl, -C(=N-CN)-NH2, -C(=NH)-NH2, -C(=NH)-NH(alkyl), -N(Y 1 )(Y 2 ), -alkylene-N(Y 1 )(Y 2 ), -C(O)N(Y 1 )(Y 2 ), and -S(O)N(Y 1 )(Y 2 )(where, Y 1 and Y 2 may be the same or different and are independently selected from the group consisting of hydrogen, alkyl, aryl, cycloalkyl, and -alkylene-aryl. A "ring system substituent" can also mean a single moiety that simultaneously replaces two available hydrogens on two adjacent carbon atoms on a ring system (one H on each carbon). Examples of such moieties include, for example, [ka] Examples include methylenedioxy, ethylenedioxy, and -C(CH3)2-, which form moieties such as

[0070] Any substituent or variable (e.g., R 5 , n, etc.) occurs more than one time in any constituent or in formula (I), its definition at each occurrence is independent of its definition at every other occurrence, unless otherwise stated.

[0071] As used herein, the term "composition" is intended to encompass products containing the specified ingredients in the specified amounts, as well as any product resulting from combining the specified ingredients in the specified amounts.

[0072] It is understood that those skilled in the art can incorporate one or more silicon (Si) atoms into the compounds of the present disclosure instead of one or more carbon atoms, to provide compounds that are chemically stable and can be easily synthesized from readily available starting materials by techniques known in the art.Carbon and silicon have different covalent bond radii, and when comparing the bonds of similar C and Si elements, the bond distance and steric configuration are different.These differences result in subtle changes in the size and shape of silicon-containing compounds compared with carbon.Those skilled in the art will understand that the difference in size and shape can result in subtle or dramatic changes in potency, solubility, lack of off-target activity, packaging properties, etc. (Diass, JO et al. Organometallics (2006) 5:1188-1198; Showell, GA et al. Bioorganic & Medicinal Chemistry Letters (2006) 16:2555-2558).

[0073] It is understood that the substituents and substitution patterns on the compounds of the present disclosure can be selected by those skilled in the art to provide compounds that are chemically stable and can be easily synthesized from readily available starting materials using techniques known in the art and the methods described below. When a substituent itself is substituted with two or more groups, it is understood that these multiple groups can be on the same carbon or on different carbons, as long as a stable structure is obtained. The phrase "optionally substituted with one or more substituents" should be understood to mean that the group in question can be unsubstituted or substituted with one or more substituents.

[0074] When optional substitution of a moiety is described (e.g., "optionally substituted"), the term means that, if substituents are present, one or more of the listed substituents for the specified substrate may be present on the substrate at the bonding position normally occupied by the default substituent normally occupying that position. For example, the default substituent on a carbon atom of an alkyl moiety is a hydrogen atom, and an optional substituent can replace the default substituent.

[0075] Pharmaceutically acceptable esters of the present compounds include the following groups: (1) ester groups in which the non-carbonyl moiety of the carboxylic acid moiety of the ester group is a straight or branched chain alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, t-butyl, sec-butyl, or n-butyl), alkoxyalkyl (e.g., methoxymethyl), aralkyl (e.g., benzyl), aryloxyalkyl (e.g., phenoxymethyl), aryl (e.g., halogen, C 1~4 Alkyl, -O-(C 1~4(1) carboxylic acid esters obtained by esterification of the hydroxy group of a hydroxyl compound selected from alkyl (phenyl optionally substituted with alkyl) or amino (phenyl optionally substituted with amino); (2) sulfonic acid esters such as alkyl- or aralkyl sulfonyl (e.g., methanesulfonyl); (3) amino acid esters (e.g., L-valyl or L-isoleucyl); (4) phosphonic acid esters, and (5) mono-, di-, or triphosphate esters. Phosphate esters are, for example, C 1~20 Alcohol or its reactive derivative, or 2,3-di(C 6~24 ) may be further esterified with acylglycerol.

[0076] One or more compounds of the present disclosure can exist in unsolvated and solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and the present disclosure is intended to encompass both solvated and unsolvated forms. "Solvate" refers to a physical association of a compound of the present disclosure with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances, a solvate is capable of isolation, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. "Solvate" encompasses both solution-phase and isolatable solvates. Non-limiting examples of solvates include ethanolates, methanolates, and the like. A "hydrate" is a solvate in which the solvent molecule is water.

[0077] One or more compounds of the present disclosure may optionally be converted into a solvate. The preparation of solvates is generally known. For example, M. Caira et al., J. Pharmaceutical Sci., 93(3), 601-611 (2004) describes the preparation of a solvate of the antifungal drug fluconazole from ethyl acetate and water. Similar preparations of solvates, hemisolvates, hydrates, etc. are described by EC van Tonder et al., AAPS PharmSciTechours., 5(1), article 12 (2004); and A.L. Bingham et al., Chem. Commun., 603-604 (2001). A typical, non-limiting process includes dissolving a compound of the invention in a desired amount of a desired solvent (organic or aqueous, or a mixture thereof) at a temperature above room temperature, cooling the solution at a rate sufficient to form crystals, which are then isolated by standard methods. Analytical techniques such as, for example IR spectroscopy, show the presence of the solvent (or water) in the crystals as a solvate (or hydrate).

[0078] Linker-payload compounds can also form salts within the scope of the present disclosure.As used herein, the term "pharmaceutically acceptable salt" or "salt" refers to a derivative in which the parent compound is modified by making its acid or base salt.Salts in solid form may exist in two or more crystal structures, and may also be in the form of hydrates.

[0079] Representative acid addition salts include acetate, ammonium, ascorbate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphorate, camphorsulfonate, fumarate, hydrochloride, hydrobromide, hydroiodide, lactate, maleate, methanesulfonate (also known as mesylate), naphthalenesulfonate, nitrate, oxalate, phosphate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate (also known as tosylate), etc. Additionally, acids generally considered suitable for forming pharmaceutically useful salts from basic pharmaceutical compounds are described, for example, in P. Stahl et al., Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. 2 nd Revised Ed. (2011) Zurich: Wiley-VCH; S. Berge et al., Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986)-33 201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and The Orange Book (Food & Drug Administration, Washington, DC, on their website), the disclosures of which are incorporated herein by reference. In one embodiment, the acid salt is an ammonium salt or a diammonium salt.

[0080] Representative base salts include ammonium salts, alkali metal salts (such as sodium, lithium, and potassium salts), alkaline earth metal salts (such as calcium and magnesium salts), salts with organic bases (e.g., organic amines) (e.g., dicyclohexylamine, t-butylamine, choline, etc.), and salts with amino acids (arginine, lysine, etc.), etc. Basic nitrogen-containing groups can be quaternized with agents such as lower alkyl halides (e.g., methyl, ethyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, and dibutyl sulfate), long chain halides (e.g., decyl chloride, bromide, and iodide, lauryl, and stearyl), aralkyl halides (e.g., benzyl and phenethyl bromides), and the like.

[0081] All such acid and base salts are intended to be pharmaceutically acceptable salts within the scope of this disclosure, and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of this disclosure.

[0082] The compounds of the present disclosure may contain one or more asymmetric centers and thus can exist as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. Depending on the nature of the various substituents on the molecule, additional asymmetric centers may exist. Each of these asymmetric centers independently produces two optical isomers, and all possible optical isomers and diastereomers in mixtures, as well as pure or partially purified compounds, are intended to be included within the scope of the present disclosure. Unless a specific stereochemistry is indicated, the present disclosure is intended to encompass all such isomeric forms of these compounds.

[0083] The independent syntheses of these diastereomers or their chromatographic separations may be achieved as known in the art by appropriate modification of the methodology disclosed herein. Their absolute stereochemistry may be determined, among other methods, by the X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing an asymmetric center of known absolute configuration.

[0084] Diastereomeric mixtures can be separated into individual diastereomers based on their physical chemical differences by methods well known to those skilled in the art, such as, for example, chromatography and / or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers, and converting the individual diastereomers into the corresponding pure enantiomers (e.g., by hydrolysis). Stereochemically pure compounds can also be prepared using chiral starting materials or salt resolution techniques. Additionally, some linker-payload compounds may be atropisomers (e.g., substituted biaryls) and are considered part of the present disclosure. Enantiomers can also be directly separated using chiral chromatographic techniques.

[0085] It is also possible that linker-payload compounds may exist in different tautomeric forms, and all such forms are encompassed within the scope of the present disclosure. For example, all keto-enol and imine-enamine forms of the compounds are included in the present disclosure.

[0086] All stereoisomers (e.g., geometric isomers, optical isomers, etc.) of the compounds (including salts, solvates, hydrates, esters, and prodrugs of the compounds, and salts, solvates, and esters of the prodrugs), such as those that may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotamer forms, atropisomers, and diastereomeric forms, are contemplated within the scope of the present disclosure. If the linker-payload compound incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are encompassed within the scope of the present disclosure.

[0087] In all cases, the compound name(s) accompany the depicted structure and are intended to capture each of the possible stereochemical arrangements for a given structural isomer based on the synthetic procedures used in its preparation. The listing of an individual stereoisomer indicates that the presented compound (e.g., "Example No.") was isolated as a single stereoisomer, and that the identity of that stereoisomer corresponds to one of the possible configurations listed. The listing of an individual stereoisomer indicates that the presented compound was isolated as a racemic or diastereomeric mixture.

[0088] Specific absolute configurations are indicated by the use of a bold wedge or dashed wedge. Unless a specific absolute configuration is indicated, the present disclosure is meant to encompass all such stereoisomeric forms of these compounds.

[0089] As used herein, when there are multiple oxygen and / or sulfur atoms present in a ring system, there cannot be any adjacent oxygen and / or sulfur atoms present in said ring system.

[0090] Individual stereoisomers of the compounds of the present disclosure may be, for example, substantially free of other isomers, or may be, for example, racemic or mixed with all other or selected stereoisomers. The chiral centers of the present disclosure may have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of terms such as "salts," "solvates," "esters," "prodrugs," and the like is intended to apply equally mutatis mutandis to enantiomers, stereoisomers, rotamers, tautomers, positional isomers, racemates, or prodrug salts, solvates, esters, and prodrugs of the compounds of the present invention.

[0091] In the compounds of formula (I), atoms may be present at their natural isotopic abundance, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number but an atomic mass or mass number different from that predominantly found in nature. The present disclosure is meant to include all suitable isotopic variations of the compounds of general formula I. For example, different isotopic forms of hydrogen (H) include protium ( 1 H) and deuterium ( 2 H). Protium is the predominant hydrogen isotope found in nature. Deuterium enrichment may provide certain therapeutic advantages, such as increased in vivo half-life or reduced dosage requirements, or may provide compounds useful as standards for characterization of biological samples. Isotopically enriched compounds of formula (I) can be prepared without undue experimentation by conventional techniques well known to those skilled in the art, or by methods analogous to those described in the schemes and examples herein using appropriate isotopically enriched reagents and / or intermediates. In one embodiment, the compound of formula (I) has one or more of its hydrogen atoms replaced with deuterium.

[0092] Polymorphic forms of the linker-payload compounds, and of the salts, solvates, hydrates, esters and prodrugs of the linker-payload compounds, are intended to be included in the present disclosure.

[0093] For all embodiments described herein, any variable not explicitly defined in the embodiment is as defined in formula (I). For each embodiment described herein, each variable is selected independently of the others, unless otherwise specified.

[0094] The following abbreviations are used below and have the following meanings: [Table 1] TIFF2025542248000029.tif59166Compounds of the present disclosure Novel linker-payload compounds (compounds of formula (I)) containing a PNU anthracycline payload are described. The compounds of formula (I) are useful as components of antibody-drug conjugates useful for the treatment and prevention of cancer. Antibody-drug conjugates comprising compounds of formula (I), compositions comprising such antibody-drug conjugates, and the use of such antibody-drug conjugates for treating or preventing cancer are also described.

[0095] Linker-payload compounds of the present disclosure In one aspect, the present disclosure provides linker-payload compounds comprising a PNU anthracycline payload and a cleavable linker, according to Formula (I) and pharmaceutically acceptable salts thereof: [ka] (In the formula, R 1 and R 2 is defined above).

[0096] In one embodiment, R 1 -XYZR 3 is selected from.

[0097] In one embodiment, R 2 is H.

[0098] In another embodiment, R 2 is a C1-C6 alkyl.

[0099] In certain embodiments, R 2 is H.

[0100] In certain embodiments, R 2 is methyl.

[0101] In one embodiment, R 1 -XYZR 3 and R 2 is H or C1-C6 alkyl.

[0102] In one embodiment, R 1 and R 2 are linked to form a 4- to 6-membered monocyclic heterocycloalkylene group, which is bonded to a ring carbon atom by -XYZR 3 is replaced by .

[0103] In another embodiment, R 1 and R 2 are joined to form a 5- to 11-membered bicyclic heterocycloalkylene group, which is bonded to a ring carbon atom by -XYZR 3 is replaced by .

[0104] In certain embodiments, R 1 and R 2 are combined, [ka] Form.

[0105] In one embodiment, R 3 but [ka] is.

[0106] In another embodiment, R 3 but [ka] is.

[0107] In another embodiment, R 3 but [ka] is.

[0108] In yet another embodiment, R 3 but [ka] is.

[0109] In certain embodiments, R 3 but [ka] is.

[0110] In another particular embodiment, R 3 but [ka] is.

[0111] In another particular embodiment, R 3 but [ka] is.

[0112] In another particular embodiment, R 3 but [ka] is.

[0113] In yet another particular embodiment, R 3 but [ka] is.

[0114] In yet another particular embodiment, R 3 but [ka] is.

[0115] In another particular embodiment, R 3 but [ka] is.

[0116] In another particular embodiment, R 3 but [ka] is.

[0117] In one embodiment, X is -(CH2) n -N(R 4 )-.

[0118] In another embodiment, X is -(CH2) n -NH-, and n is 1 or 2.

[0119] In another embodiment, X is -(CH2) n -N(CH3)-, where n is 1 or 2.

[0120] In one embodiment, X is -NH-.

[0121] In another embodiment, X is -R 5 -NH-.

[0122] In one embodiment, X is -(CH2) n -N(R 4 )-C(O)R 5 -It is.

[0123] In another embodiment, X is -(CH2) n -N(R 4 )-(C1-C6 alkylene)-NH(R 4 )-.

[0124] In another embodiment, X is -(CH2) n -N(R 4 )-C(O)O(CH2) n -R 5 -N(R 4 )-.

[0125] In another embodiment, X is —(CH 2 ) 3 —NH—(CH 2 ) 3 —NH—.

[0126] In another embodiment, X is a 5- or 6-membered monocyclic heterocycloalkylene.

[0127] In certain embodiments, X is [ka] is.

[0128] In another embodiment, X is -(CH2) n -R 5 -It is.

[0129] In certain embodiments, X is —CH—R 5 -It is.

[0130] In another particular embodiment, X is [ka] is.

[0131] In yet another embodiment, X is -(CH2) n -R 5 -NH(R 4 )-.

[0132] In another embodiment, X is a 5-11 membered bicyclic spirocyclic heterocycloalkylene.

[0133] In certain embodiments, X is [ka] is.

[0134] In yet another embodiment, X is -(CH2) n -R 5 -NHC(O)OR 5 NH(R 4 )-.

[0135] In a further embodiment, X is -(CH2) n -NHC(O)O-(CH2) n -R 5 -NH(R 4 )

[0136] In another embodiment, X is -(CH2) n -R 5 -OC(O)-NH(R 4 )-(C1-C6 alkylene)-NH(R 4 )-.

[0137] In one embodiment, X is —R 5 -NHC(O)CH2OCH2N(R 4 )C(O)CHN(R 4 )-.

[0138] In certain embodiments, X is [ka] is.

[0139] In another embodiment, X is -(CH2) n -N(R 4 )C(O)NH-N(R 4 )-.

[0140] In certain embodiments, X is —CH 2 CH 2 NHC(O)NHNH—.

[0141] In another embodiment, X is -R 5 -N(R 4 )C(O)CHN(R 4 )-.

[0142] In certain embodiments, X is [ka] is.

[0143] In yet another embodiment, X is -(CH2) n -N(R 4 )-(CH2) n -N(R 4 )-.

[0144] In certain embodiments, X is —CH 2 CH 2 NH—CH 2 CH 2 NH—.

[0145] In a further embodiment, -R 5 -O-CH2N(R 4 )-.

[0146] In certain embodiments, X is [ka] is.

[0147] In one embodiment, Y is a bond.

[0148] In another embodiment, Y is —C(O)C(R 4 )(R 6 )NH-.

[0149] In another embodiment, Y is —C(O)C(R 4 )(R 6 )NH- and R 6 is H or isopropyl, and R 4 is H or methyl.

[0150] In one embodiment, Z is —C(O)CH(R 7 )NHC(O)CH(R 8 )NH- and R 7 and R 8are each independently selected from H, methyl, isopropyl, and —CH 2 CH 2 CH 2 NHC(O)NH 2 .

[0151] In one embodiment, R 5 C6~C 10 It is aryl.

[0152] In another embodiment, R 5 is a C3-C7 monocyclic cycloalkylene.

[0153] In another embodiment, R 5 C5~C 11 It is a bicyclic cycloalkylene.

[0154] In another embodiment, R 5 is -NHC(O)NH-.

[0155] In another embodiment, R 5 is -NH-NH-.

[0156] In certain embodiments, R 5 -NH-NH-, -NHC(O)NH-, [ka] is selected from.

[0157] In one embodiment, X is -(CH) n -N(R 4 )-, -R 5 -N(R 4 )-, -(CH2) n -N(R 4 )-C(O)R 5 -, -(CH2) n -N(R 4 )-(C1-C6 alkylene)-N(R 4 )-, -(CH2) n -R 5 -N(R 4 )-, -(CH2) n -R 5 -, -(CH2) n -R5 -NHC(O)O-(CH2) n R 5 -N(R 4 )-, -(CH2) n -NHC(O)O-(CH2) n -R 5 -N(R 4 )-, -(CH2) n -R 5 -OC(O)-N(R 4 )-(C1-C6 alkylene)-N(R 4 )-, -R 5 -NHC(O)CH2OCH2N(R 4 )C(O)CHN(R 4 )-, -R 5 -N(R 4 )C(O)CHN(R 4 )-, -(CH2) n -N(R 4 )-CH2-N(R 4 )-, and -R 5 -O-CH2N(R 4 )-selected from; R 4 each occurrence of is independently H, methyl, or Fmoc; R 5 each occurrence of is -NH-NH-, -NHC(O)NH, [ka] are independently selected from

[0158] In one embodiment, -XYZR 3 The base is [ka] TIFF2025542248000053.tif230165 TIFF2025542248000054.tif214166 Selected from TIFF2025542248000055.tif173165.

[0159] In one embodiment, the compound of formula (I) is a compound of formula (I'): [ka] and pharmaceutically acceptable salts thereof, wherein R 2 is H or C1-C6 alkyl, R 1 -XYZR 3 Selected from; R 2 is H or C1-C6 alkyl, or R 1 and R 2 are joined to form a 4- to 6-membered monocyclic heterocycloalkylene group or a 5- to 11-membered bicyclic spirocyclic heterocycloalkylene group, each of which is bonded to a ring carbon atom by -XYZR 3 is replaced by; R 3 teeth, [ka] Selected from; X is -(CH2) n -N(R 4 )-, -R 5 -N(R 4 )-, -(CH2) n -N(R 4 )-C(O)R 5 -, -(CH2) n -N(R 4 )-(C1-C6 alkylene)-N(R 4 )-, 5- or 6-membered monocyclic heterocycloalkylene, -(CH2) n -R 5 -N(R 4 )-, 5-11 membered bicyclic spirocyclic heterocycloalkylene, -(CH2) n -R 5 -NHC(O)O-(CH2) n R 5 -N(R 4 )-, -(CH2) n -NHC(O)O-(CH2) n -R 5 -N(R 4 )-, and -(CH2) n -R 5-OC(O)-N(R 4 )-(C1-C6 alkylene)-N(R 4 )-selected from; Y is a bond or -C(O)C(R 4 )(R 6 )NH-; Z is -C(O)CH(R 7 )NHC(O)CH(R 8 )NH-; R 4 is H, C 1~6 alkyl, or [ka] and; R 5 is C6~C 10 Arylene, 4-6 membered monocyclic heterocycloalkylene, 5-11 membered bicyclic spirocyclic heterocycloalkylene, C3-C7 monocyclic cycloalkylene, C5-C 11 bicyclic cycloalkylene, and -NHC(O)NH-, wherein said 4-6 membered monocyclic heterocycloalkylene may be optionally substituted on a ring carbon atom with an oxo group; R 6 , R 7 and R 8 are each independently selected from H, —CHCHCHNHC(O)NH, and a naturally occurring amino acid side chain; m is 0, 1 or 2; each occurrence of n is independently an integer from 0 to 4. is.

[0160] In one embodiment, the compound of formula (I) is of formula (II): [ka] (In the formula, R 2 , X, Y, Z, and m are as described herein for compounds of formula (I). It is expressed as:

[0161] In another embodiment, the compound of formula (I) is of formula (III): [ka] wherein X, Y, Z, and m are as described herein; and A is a 4- or 6-membered monocyclic heterocycloalkylene group. It is expressed as:

[0162] In another embodiment, the compound of formula (I) is of formula (IV): [ka] (In the formula, R 2 , X, Y, Z, and m are as described herein. It is expressed as:

[0163] In yet another embodiment, the compound of formula (I) has formula (V): [ka] wherein X, Y, Z, and m are as described herein; and A is a 4- or 6-membered monocyclic heterocycloalkylene group. It is expressed as:

[0164] In a further embodiment, the compound of formula (I) is of formula (VI): [ka] (In the formula, R 2 , X, Y, Z, and m are as described herein. It is expressed as:

[0165] In another embodiment, the compound of formula (I) is of formula (VII): [ka] wherein X, Y, Z, and m are as described herein; and A is a 4- or 6-membered monocyclic heterocycloalkylene group. It is expressed as:

[0166] In another embodiment, the compound of formula (I) is of formula (VIII): [ka] (In the formula, R 2 , R 9 , X, Y, Z, and m are as described herein. It is expressed as:

[0167] In another embodiment, the compound of formula (I) is of formula (IX): [ka] (In the formula, R 9 , X, Y, Z, and m are as described herein; and A is a 4- or 6-membered monocyclic heterocycloalkylene group. It is expressed as:

[0168] In another embodiment, it may be desirable to synthesize the linker prior to conjugation to the drug or targeting moiety to provide an antibody-drug conjugate. In such an embodiment, the linker compound may act as an intermediate compound. Exemplary linkers of the present disclosure include, but are not limited to, the following linker fragments (L-1 to L-12), where the attachment point of the payload to the linker is: [ka] Represented by: [Table 2] The linker fragments L-1 to L-12 can be linked to payloads using the following reactants L-1' to L-12', respectively, where Q is -OH or -Cl: [Table 3] TIFF2025542248000071.tif95165 In one aspect, the present disclosure provides linker-payload compounds having structural formulas (X)-(XXI), where D is an anticancer drug: [Table 4] TIFF2025542248000073.tif166166In one embodiment, the disclosure provides a linker-payload compound comprising a linker moiety selected from L-1, L-2, L-1, L-4, L-5, L-6, L-7, L-8, L-9, L-10, L-11, and L-12 conjugated to an anticancer drug having a reactive -OH group.

[0169] In one embodiment, the present disclosure provides a linker-payload compound comprising a linker moiety selected from L-1, L-2, L-1, L-4, L-5, L-6, L-7, L-8, L-9, L-10, L-11, and L-12 conjugated to an anticancer drug that is a PNU anthracycline.

[0170] In certain embodiments, the present disclosure provides a compound comprising: (i) a linker (L) selected from L-1, L-2, L-1, L-4, L-5, L-6, L-7, L-8, L-9, L-10, L-11, and L-12; and (ii) a PNU anthracycline compound of formula (XXII): [ka] or a pharmaceutically acceptable salt thereof, R 1’ is H or C 1~6 is alkyl; R 2’ is a linker selected from linkers L-1 to L-12; Alternatively, R 1 and R 2taken together with the common nitrogen atom to which they are each bonded, combine to form (i) a 3- to 7-membered monocyclic heterocycloalkyl group, (ii) a 5- to 11-membered bridged bicyclic heterocycloalkyl group, (iii) a 5- to 11-membered fused bicyclic heterocycloalkyl group, or (iv) a 5- to 11-membered spirocyclic heterocycloalkyl group, each of which may optionally and independently be selected from the group consisting of one or more R A a linker selected from linkers L-1 to L-12, which may be substituted with a group, attached to a ring atom of (i), (ii), (iii), or (iv); R A Each occurrence of C 1~6 Alkyl, halo, -CN, -OR 3 , -N(R 3 )2, C1-C6 aminoalkyl, -(C1-C6 alkylene)-NHC(O)-(C1-C6 aminoalkyl), -O-(C 1~6 and -(C-C alkylene)-(3- to 7-membered monocyclic heterocycloalkyl), wherein the 3- to 7-membered monocyclic heterocycloalkyl group is optionally and independently selected from one or more R B optionally substituted with a group; R B Each occurrence of C 1~6 Alkyl, halo, -CN, -OR 3 , -N(R 3 )2, C1-C6 aminoalkyl, -O-(C 1~6 alkyl), -NHC(O)-(C1-C6 aminoalkyl), -(C1-C6 alkylene)-NHC(O)-(C1-C6 aminoalkyl), C 1~6 Alkyl, halo, -CN, -OR 3 , -N(R 3 )2, -O-(C 1~6 and a payload that is independently selected from —NHC(O)CHOH).

[0171] The payload of formula (XVI) can be linked to linkers of formula L-1, L-2, L-1, L-4, L-5, L-6, L-7, L-8, L-9, L-10, L-11, and L-12 using methods described herein or known in synthetic organic chemistry.

[0172] In one embodiment, the linker-payload compounds of the present disclosure are in substantially purified form.

[0173] Non-limiting examples of linker-payload compounds of Formula (I) include Examples 1-36, and pharmaceutically acceptable salts thereof. [Table 5] TIFF2025542248000076.tif194166 TIFF2025542248000077.tif187165 TIFF2025542248000078.tif203166 TIFF2025542248000079.tif233165 TIFF2025542248000080.tif230166 TIFF2025542248000081.tif219165 TIFF2025542248000082.tif239166 TIFF2025542248000083.tif224165 TIFF2025542248000084.tif246165 TIFF2025542248000085.tif63165ADC disclosed in this document The linker-payload compounds of the present disclosure have utility for conjugation to antibodies or other targeting moieties to generate antibody-drug conjugates or other targeting ligand conjugates for oncology indications. Thus, in one aspect, the present disclosure provides antibody-drug conjugates ("ADCs of the present disclosure") comprising a linker-payload of the present disclosure linked to a ligand (e.g., an antibody).

[0174] Accordingly, certain embodiments of the present disclosure provide compounds of structural formula (XXIII): [ka] (In the formula, R 2 , R 9 , X, Y, and Z are described herein above for compounds of formula (I); p is an integer from 1 to 8; L is a ligand, e.g., an antibody or other targeting moiety; R 3’ teeth, [ka] (wherein ** is R 3’ (indicating the point of attachment of to L) (selected from The antibody-drug conjugate is represented by the formula:

[0175] In one embodiment, L is an antibody and R 3’ is attached to the sulfur atom of a cysteine ​​residue of the antibody.

[0176] Another embodiment of the present disclosure is a compound of structural formula (XXIV): [ka] wherein X, Y, and Z are as described herein; A is a 4- or 6-membered monocyclic heterocycloalkylene group; p is an integer from 1 to 8; L is a ligand, e.g., an antibody or other targeting moiety; R 3’ teeth, [ka] (wherein ** is R 3’ (indicating the point of attachment of to L) (selected from The antibody-drug conjugate is represented by the formula:

[0177] In one embodiment, L is an antibody and R 3’ is attached to the sulfur atom of a cysteine ​​residue of the antibody.

[0178] Another embodiment of the present disclosure is a compound of structural formula (XXV): [ka] (In the formula, R 2 , X, Y, Z, and m are as described herein; p is an integer from 1 to 8; and L is a ligand, e.g., an antibody or other targeting moiety. The antibody-drug conjugate is represented by the formula:

[0179] Yet another embodiment of the present disclosure is a compound of structural formula (XXVI): [ka] wherein X, Y, Z, and m are as described herein; A is a 4- or 6-membered monocyclic heterocycloalkylene group; p is an integer from 1 to 8; and L is a ligand, e.g., an antibody or other targeting moiety. The antibody-drug conjugate is represented by the formula:

[0180] Further embodiments of the present disclosure include compounds of structural formula (XXVII): [ka] (In the formula, R 2 , X, Y, Z, and m are as described herein; p is an integer from 1 to 8; and L is a ligand, e.g., an antibody or other targeting moiety. The antibody-drug conjugate is represented by the formula:

[0181] Another embodiment of the present disclosure is a compound of structural formula (XXVIII): [ka] wherein X, Y, Z, and m are as described herein; A is a 4- or 6-membered monocyclic heterocycloalkylene group; p is an integer from 1 to 8; and L is a ligand, e.g., an antibody or other targeting moiety. The antibody-drug conjugate is represented by the formula:

[0182] Yet another embodiment of the present disclosure is a compound of structural formula (XXIX): [ka] (In the formula, R 2 , X, Y, Z, and m are as described herein; p is an integer from 1 to 8; and L is a ligand, e.g., an antibody or other targeting moiety. The antibody-drug conjugate is represented by the formula:

[0183] Another embodiment of the present disclosure is a compound of structural formula (XXX): [ka] wherein X, Y, Z, and m are as described herein; A is a 4- or 6-membered monocyclic heterocycloalkylene group; p is an integer from 1 to 8; and L is a ligand, e.g., an antibody or other targeting moiety. The antibody-drug conjugate is represented by the formula:

[0184] Another embodiment of the present disclosure is a compound of structural formula (XXXI): [ka] (In the formula, R 2 , R 9 , X, Y, and Z are as described herein; p is an integer from 1 to 8; and L is a ligand, e.g., an antibody or other targeting moiety. The antibody-drug conjugate is represented by the formula:

[0185] Another embodiment of the present disclosure is a compound of structural formula (XXXII): [ka] (In the formula, R 9 , X, Y, and Z are as described herein; A is a 4- or 6-membered monocyclic heterocycloalkylene group; p is an integer from 1 to 8; and L is a ligand, e.g., an antibody or other targeting moiety. The antibody-drug conjugate is represented by the formula:

[0186] In one embodiment, for compounds of Formulae (XXIII)-(XXXII), L is an antibody and the linker is attached to the sulfur atom of a cysteine ​​group of said antibody.

[0187] In one embodiment, the antibody-drug conjugate of the present disclosure comprises the following compounds having structural formulas (XXXIII)-(LVII): [Table 6] TIFF2025542248000099.tif244166 TIFF2025542248000100.tif233165 TIFF2025542248000101.tif247166 TIFF2025542248000102.tif223165 TIFF2025542248000103.tif186165 TIFF2025542248000104.tif206166 TIFF2025542248000105.tif219165 TIFF2025542248000106.tif160166 (wherein L is a ligand and p is an integer from 0 to 8).

[0188] In one embodiment, the antibody drug conjugates of the present disclosure are in substantially purified form.

[0189] Non-limiting examples of antibody-drug conjugates of the present disclosure include Examples 37-72 and pharmaceutically acceptable salts thereof (wherein L is sacituzumab (S365C) ​​and p is an integer from 1 to 8). [Table 7] TIFF2025542248000108.tif202165 TIFF2025542248000109.tif201166 TIFF2025542248000110.tif223165 TIFF2025542248000111.tif201165 TIFF2025542248000112.tif228166 TIFF2025542248000113.tif201165 TIFF2025542248000114.tif188165 TIFF2025542248000115.tif198165 TIFF2025542248000116.tif242165 TIFF2025542248000117.tif244165 TIFF2025542248000118.tif77165Other embodiments include: (a) A composition comprising a mixture of ADCs of the disclosure, wherein the DAR of the composition is a decimal number between 0 and 8.

[0190] (b) A pharmaceutical composition comprising an effective amount of an ADC of the disclosure and a pharmaceutically acceptable carrier.

[0191] (c) The pharmaceutical composition of (a), further comprising a second therapeutic agent selected from the group consisting of anticancer agents.

[0192] (d) The pharmaceutical composition of (b), wherein the anticancer agent is an anti-human PD-1 antibody (or an antigen-binding fragment thereof).

[0193] (e) A pharmaceutical combination comprising (i) an ADC of the present disclosure and (ii) a second therapeutic agent selected from the group consisting of anticancer agents, wherein the ADC of the present disclosure and the second therapeutic agent are each used in an amount that renders the combination effective to inhibit cancer cell replication or treat cancer and / or reduce the likelihood of cancer or the severity of its symptoms.

[0194] (f) The combination of (e), wherein the second therapeutic agent is an anti-human PD-1 antibody (or an antigen-binding fragment thereof).

[0195] (g) A method of inhibiting cancer cell replication in a subject in need thereof, comprising the step of administering to the subject an effective amount of an ADC of the disclosure.

[0196] (h) A method of treating cancer and / or reducing the likelihood of or the severity of symptoms of cancer in a subject in need thereof, comprising administering to the subject an effective amount of an ADC of the disclosure.

[0197] (i) The method of (h), wherein an ADC of the present disclosure is administered in combination with an effective amount of at least one second therapeutic agent selected from the group consisting of anticancer agents.

[0198] (j) The method of (i), wherein the second therapeutic agent is an anti-human PD-1 antibody (or an antigen-binding fragment thereof).

[0199] (k) A method of inhibiting cancer cell replication in a subject in need thereof, comprising administering to the subject a composition of (a); a pharmaceutical composition of (b), (c) or (d), or a combination of (e) or (f).

[0200] (l) A method of treating cancer and / or reducing the likelihood or severity of symptoms of cancer in a subject in need thereof, comprising administering to the subject: (a) a composition; (b), (c) or (d) a pharmaceutical composition; or (e) or (f) a combination.

[0201] Also described herein are ADCs of the present disclosure for use in (i), (ii) as a medicament for, or (iii) for preparing a medicament for (a) a medicine; (b) inhibiting cancer cell replication; or (c) treating cancer and / or reducing the likelihood or severity of symptoms of cancer. In these uses, the ADCs of the present disclosure may optionally be used in combination with one or more additional therapeutic agents selected from anti-cancer agents.

[0202] It should further be understood that the composition and method embodiments provided as (a) through (k) above are understood to include all embodiments of the compounds, including such embodiments as a result of combinations of embodiments.

[0203] Ligand In one embodiment, the linker-payload compounds of the present disclosure can be conjugated to a ligand, such as an antibody, to provide an antibody-drug conjugate (ADC of the present disclosure), in which the ligand is linked to the linker via a bond formed between a moiety on the ligand and either a maleimide group or a sulfone-substituted pyridyl group on the linker.

[0204] The ligand can be any moiety with a free sulfur atom, including, but not limited to, an antibody, a protein, a peptide, a polypeptide, or an engineered antibody modified to provide a free cysteine. This embodiment is realized when the ligand is an antibody, preferably an intact antibody. The ligand acts to target and present drugs to a specific target cell population with which the ligand interacts. Suitable ligands include, for example, antibodies, including full-length antibodies and their antigen-binding fragments, interferons, lymphokines, hormones, growth factors and colony-stimulating factors, vitamins, nutrient transport molecules (such as, but not limited to, transferrin), or any other cell-binding molecule or substance, including small molecules and peptides. The ligand can also be, for example, a non-antibody protein targeting agent.

[0205] Where the conjugate comprises a non-immunoreactive protein, polypeptide, or peptide ligand instead of an antibody, useful non-immunoreactive protein, polypeptide, or peptide ligands include, but are not limited to, transferrin, epidermal growth factor ("EGF"), bombesin, gastrin, gastrin-releasing peptide, platelet-derived growth factor, IL-2, IL-6, transforming growth factors ("TGFs"), such as TGF-α and TGF-β, vaccinia growth factor ("VGF"), insulin and insulin-like growth factors I and II, somatostatin, lectins, and apoproteins derived from low-density lipoproteins.

[0206] Particularly preferred ligands are antibodies, including intact antibodies. Indeed, in any of the embodiments described herein, the ligand can be an antibody. Useful polyclonal antibodies are heterogeneous populations of antibody molecules derived from the serum of immunized animals. Useful monoclonal antibodies are homogeneous populations of antibodies against a specific antigenic determinant (e.g., cancer cell antigens, viral antigens, microbial antigens, proteins, peptides, carbohydrates, chemicals, nucleic acids, or fragments thereof). Monoclonal antibodies (mAbs) against an antigen of interest can be prepared by using any technique known in the art that provides for the production of antibody molecules by continuous cell lines in culture. In one embodiment, the ligand is an antibody and is linked to a linker via a cysteine ​​group.

[0207] Additionally, recombinant antibodies, such as chimeric antibodies and humanized monoclonal antibodies, containing both human and non-human portions, are useful antibodies and can be produced using standard recombinant DNA techniques. Chimeric antibodies are molecules in which different portions are derived from different animal species, such as those having a variable region derived from a mouse monoclonal and a human immunoglobulin constant region (see, e.g., U.S. Pat. Nos. 4,816,567 and 4,816,397, the entire contents of which are incorporated herein by reference). Humanized antibodies are antibody molecules derived from species other than human, having one or more complementarity-determining regions (CDRs) from a non-human species and a framework region derived from a human immunoglobulin molecule (see, e.g., U.S. Pat. No. 5,585,089, the entire contents of which are incorporated herein by reference). Such chimeric antibodies and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art, for example, using the methods described in WO 87 / 02671 and EP 0184187, the entire contents of which are incorporated herein by reference.

[0208] Completely human antibodies are particularly desirable and can be produced using transgenic mice that are incapable of expressing endogenous immunoglobulin heavy and light chain genes, but which can express human heavy and light chain genes.

[0209] Antibodies include analogs and derivatives modified by the covalent attachment of any type of molecule, so long as the covalent attachment allows the antibody to retain its antigen-binding immunospecificity. For example, but not limited to, antibody derivatives and analogs include those further modified by, for example, glycosylation, acetylation, PEGylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, conjugation to cellular antibodies or other proteins, etc. Any of a number of chemical modifications can be performed by known techniques, including, but not limited to, specific chemical cleavage, acetylation, formylation, metabolic synthesis in the presence of tunicamycin, etc. Furthermore, analogs or derivatives can contain one or more unnatural amino acids.

[0210] In certain embodiments, known antibodies for treating cancer can be used.Antibodies immunospecific for cancer cell antigens can be obtained commercially or can be produced by any method known to those skilled in the art, such as recombinant expression technology.The nucleotide sequence encoding antibodies immunospecific for cancer cell antigens can be obtained, for example, from GenBank database or similar databases, literature publications, or by routine cloning and sequencing.

[0211] In another specific embodiment, antibodies for treating autoimmune diseases are used in accordance with the compositions and methods of the present disclosure. Antibodies immunospecific for antigens of cells responsible for the production of autoimmune antibodies can be obtained from any organization (e.g., university scientists or companies) or can be produced by any method known to those skilled in the art, such as, for example, chemical synthesis or recombinant expression techniques.

[0212] In other embodiments, it may be desirable to conjugate a component of the linker to a ligand (e.g., an antibody) before attaching the payload component of the antibody-drug conjugate. For example, in embodiments in which a thiol-containing substituent, such as cysteine, is used to attach the payload component, it may be desirable to conjugate a component of the linker to a ligand (e.g., an antibody) before attaching the payload component of the antibody-drug conjugate.

[0213] Uses of the Compounds of the Disclosure As an intermediate for synthesizing ADCs The linker-payload compounds of the present disclosure are useful as components of antibody-drug conjugates. In one embodiment, the linker-payload compounds of the present disclosure can be conjugated to a ligand (i.e., an antibody or antibody fragment) to produce an ADC of the present disclosure. The maleimide group or sulfone-substituted pyridyl group of the linker-payload compounds of the present disclosure can serve as a conjugation handle and attachment point for the ligand to the linker-payload compounds of the present disclosure. In certain embodiments, an antibody is linked to the linker-payload compounds of the present disclosure via the sulfur atom of a cysteine ​​residue on the antibody.

[0214] In certain embodiments, one or more engineered cysteine ​​groups on an antibody are reduced to provide free thiol group(s), which can then undergo a conjugation reaction with a maleimide group or a sulfone-substituted pyridyl group on a linker-payload compound of the present disclosure, thereby attaching the antibody to the linker moiety of the linker-payload compound of the present disclosure and forming an ADC of the present disclosure.

[0215] To treat and prevent cancer As noted above, additional embodiments of the present disclosure each relate to methods of treating a disease, disorder, or condition, or one or more symptoms thereof ("indications"), comprising administering a therapeutically effective amount of an ADC of the present disclosure, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising said compound or salt thereof, to a subject in need of such treatment.

[0216] One such embodiment is a method for treating breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, stomach cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), colon cancer, rectal cancer, colorectal cancer, leukemia (e.g., acute lymphocytic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, and the like) in a subject in need thereof.

[0013] In one embodiment, the present invention provides a method for treating or preventing a cancer selected from: myeloid leukemia, bone cancer, skin cancer, thyroid cancer, pancreatic cancer, and lymphoma (e.g., Hodgkin's lymphoma, non-Hodgkin's lymphoma, or recurrent anaplastic large cell lymphoma), comprising administering to a subject in need of such treatment a therapeutically effective amount of an ADC of the disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising said compound, salt or solvate thereof. In one such embodiment, the subject is human.

[0217] Another aspect of the present disclosure relates to a method for treating and / or preventing tumors, comprising administering to a patient in need thereof a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising a compound according to the present disclosure.

[0218] Combination therapy The present method also contemplates combination with additional therapeutic agents. For example, the combination of the ADC of the present disclosure with a PPAR-γ (i.e., PPAR-gamma) agonist and a PPAR-δ (i.e., PPAR-delta) agonist is useful in the treatment of certain malignant tumors. PPAR-γ and PPAR-δ are nuclear peroxisome proliferator-activated receptors γ and δ. PPAR-γ agonists have been shown to inhibit the angiogenic response to VEGF in vitro; both troglitazone and rosiglitazone maleate inhibit the development of retinal neovascularization in mice (Arch. Ophthamol. 2001;119:709-717). Examples of PPAR-γ agonists and PPAR-γ / α agonists include thiazolidinediones (such as DRF2725, CS-011, troglitazone, rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate, GW2570, SB219994, AR-H039242, JTT-501, MCC-555, GW2331, GW409544, NN2344, KRP297, NP0110, DRF4158, NN622, GI262570, PNU182716, and DRF5529. 26, 2-[(5,7-dipropyl-3-trifluoromethyl-1,2-benzisoxazol-6-yl)oxy]-2-methylpropionic acid (disclosed in U.S. Patent Application No. 09 / 782,856) and 2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchroman-2-carboxylic acid (disclosed in U.S. Patent Application Nos. 60 / 235,708 and 60 / 244,697), or a pharmaceutically acceptable salt thereof.

[0219] Another embodiment of the present disclosure is the use of the ADC of the present disclosure in combination with gene therapy to treat cancer. For a review of genetic strategies for cancer treatment, see Hall et al. (Am.J.Hum.Genet.61:785-789,1997) and Kufe et al. (Cancer Medicine, 5th Ed, pp 876-889, BC Decker, Hamilton 2000). Gene therapy can be used to deliver any tumor suppressor gene. Examples of such genes include, but are not limited to, p53, which can be delivered via recombinant virus-mediated gene transfer (see, e.g., U.S. Patent No. 6,069,134), uPA / uPAR antagonists (“Adenovirus-Mediated Delivery of a uPA / uPAR Antagonist Suppresses Angiogenesis-Dependent Tumor Growth and Dissemination in Mice,” Gene Therapy, August 1998;5(8):1105-13), and interferon gamma (J. Immunol. 2000;164:217-222).

[0220] The ADCs of the disclosure may also be administered in combination with inhibitors of intrinsic multidrug resistance (MDR), particularly MDR associated with high levels of transporter protein expression. Such MDR inhibitors include inhibitors of P-glycoprotein (P-gp), such as LY335979, XR9576, OC144-093, R101922, VX853, and PSC833 (valspodar), or pharmaceutically acceptable salts thereof.

[0221] The ADCs of the present disclosure may also be administered together with immune enhancing drugs such as levamisole, isoprinosine and Zadaxin, or pharmaceutically acceptable salts thereof.

[0222] The ADCs of the present disclosure may also be useful to treat or prevent cancer in combination with P450 inhibitors, including xenobiotics, quinidine, tyramine, ketoconazole, testosterone, quinine, metyrapone, caffeine, phenelzine, doxorubicin, troleandomycin, cyclobenzaprine, erythromycin, cocaine, furafylline, cimetidine, dextromethorphan, ritonavir, indinavir, amprenavir, diltiazem, terfenadine, verapamil, cortisol, itraconazole, mibefradil, nefazodone, and nelfinavir, or pharmaceutically acceptable salts thereof.

[0223] The ADCs of the present disclosure also include cyclosporine A, PSC833, GF120918, cremophorEL, fumitremorgin C, Ko132, Ko134, Iressa, imatinib mesylate, EKI-785, Cl1033, novobiocin, diethylstilbestrol, tamoxifen, resperpin, VX-710, tryprostatin A, flavonoids, ritonavir, saquinavir, nelfinavir, omeprazole, quinidine, verapamil, thiazolinone ... and cyclosporine, terfenadine, ketoconazole, nifidepine, FK506, amiodarone, XR9576, indinavir, amprenavir, cortisol, testosterone, LY335979, OC144-093, erythromycin, vincristine, digoxin, and talinolol, or a pharmaceutically acceptable salt thereof, may be useful for treating or preventing cancer.

[0224] The ADCs of the present disclosure may also be useful in combination with bisphosphonates, including, but not limited to, etidronate (Didronel), pamidronate (Aredia), alendronate (Fosamax), risedronate (Actonel), zoledronate (Zometa), ibandronate (Boniva), incadronate or cimadronate, clodronate, EB-1053, minodronate, neridronate, piridronate, and tiludronate (including any and all pharmaceutically acceptable salts, derivatives, hydrates, and mixtures thereof), to treat or prevent cancer, including bone cancer.

[0225] The ADCs of the present disclosure may also be useful in treating or preventing breast cancer in combination with aromatase inhibitors, including, but not limited to, anastrozole, letrozole, and exemestane, or pharmaceutically acceptable salts thereof.

[0226] The ADCs of the present disclosure may also be useful in treating or preventing cancer in combination with siRNA therapeutics.

[0227] The ADCs of the present disclosure may also be administered in combination with a gamma-secretase inhibitor and / or an inhibitor of NOTCH signaling, such as those described in WO 01 / 90084, WO 02 / 30912, WO 01 / 70677, WO 03 / 013506, WO 02 / 36555, WO 03 / 093252, WO 03 / 093264, WO 03 / 093251, WO 03 / 093253, WO 2004 / 039800, WO 2004 / 039370, WO 2005 / 030731, WO 2005 / 014553, U.S. Patent Application No. and WO 02 / 47671 (including LY-450139), or a pharmaceutically acceptable salt thereof.

[0228] In one embodiment, specific anti-cancer agents useful in the present combination therapy include pembrolizumab (Keytruda®); abarelix (Plenaxis depot®); aldesleukin (Prokine®); aldesleukin (Proleukin®); alemtuzumab (Campath®); alitretinoin (Panretin®); allopurinol (Zyloprim®); altretamine (Hexalen®); amifostine (Ethyol®); anastrozole (Arimidex®); arsenic trioxide (Trisenox®); asparaginase (Elspar®); azacitidine (Vidaza®); Registered trademark); bevacizumab (Avastin®); bexarotene capsules (Targretin®); bexarotene gel (Targretin®); bleomycin (Blenoxane®); bortezomib (Velcade®); busulfan intravenous (Busulfex®); busulfan oral (Myleran®); calsterone (Methosarb®); capecitabine (Xeloda®); carboplatin (Paraplatin®); carmustine (BCNU®, BiCNU®); carmustine (Gliadel®); carmustine implant (Gliadel®) Wafer®; celecoxib (Celebrex®); cetuximab (Erbitux®); chlorambucil (Leukeran®); cisplatin (Platinol®); cladribine (Leustatin®, 2-CdA®); clofarabine (Clolar®); cyclophosphamide (Cytoxan®, Neosar®); cyclophosphamide (Cytoxan Injection®); cyclophosphamide (Cytoxan Tablet®); cytarabine (Cytosar-U®); liposomal cytarabine (DepoCyt®);Dacarbazine (DTIC-Dome®); Dactinomycin, Actinomycin D (Cosmegen®); Darbepoetin alfa (Aranesp®); Liposomal daunorubicin (DanuoXome®); Daunorubicin, daunomycin (Daunorubicin®); Daunorubicin, daunomycin (Cerubidine®); Denileukin diftitox (Ontak®); Dexrazoxane (Zinecard®); Docetaxel (Taxotere®); Doxorubicin (Adriamycin PFS®); Doxorubicin (Adriamycin®, Rubex®); Doxorubicin (Adriamycin PFS®) Injection®); liposomal doxorubicin (Doxil®); dromostanolone propionate (Dromostanolone®); dromostanolone propionate (Masterone injection®); Elliott's B solution (Elliott's B Solution®); epirubicin (Ellence®); epoetin alfa (epogen®); erlotinib (Tarceva®); estramustine (Emcyt®); etoposide phosphate (Etopophos®); etoposide, VP-16 (Vepesid®); exemestane (Aromasin®); filgrastim (Neupogen®); floxuridine (intra-arterial) (FUDR®); fludarabine (Fludara®); fluorouracil, 5-FU (Adrucil®); fulvestrant (Faslodex®); gefitinib (Iressa®); gemcitabine (Gemzar®); gemtuzumab ozogamicin (Mylotarg®); goserelin acetate (Zoladex®) Implant®); goserelin acetate (Zoladex®); histrelin acetate (Histrelin implant®); hydroxyurea (Hydrea®);Ibritumomab tiuxetan (Zevalin®); idarubicin (Idamycin®); ifosfamide (IFEX®); imatinib mesylate (Gleevec®); interferon alpha-2a (Roferon A®); interferon alpha-2b (Intron®) A®); irinotecan (Camptosar®); lenalidomide (Revlimid®); letrozole (Femara®); leucovorin (Wellcovorin®, Leucovorin®); leuprolide acetate (Eligard®); levamisole (Ergamisol®); lomustine CCNU (CeeBU®); mechlorethamine, nitrogen mustard (Mustargen®); megestrol acetate (Megace®); melphalan, L-PAM (Alkeran®); mercaptopurine, 6-MP (Purinethol®); mesna (Mesnex®); mesna (Mesnex®) tabs®); methotrexate (Methotrexate®); methoxsalen (Uvadex®); mitomycin C (Mutamycin®); mitotane (Lysodren®); mitoxantrone (Novantrone®); nandrolone phenylpropionate (Durabolin-50®); nelarabine (Arranon®); nofetumomab (Verluma®); oprelvekin (Neumega®); oxaliplatin (Eloxatin®); paclitaxel (Paxene®); paclitaxel (Taxol®); paclitaxel protein-bound particles (Abraxane®); palifermin (Kepivance®); pamidronate (Aredia®); pegademase (Adagen®) Bovine®); pegaspargase (Oncaspar®); pegfilgrastim (Neulasta®);Pemetrexed disodium (Alimta®); pentostatin (Nipent®); pipobroman (Vercyte®); plicamycin, mithramycin (Mithracin®); porfimer sodium (Photofrin®); procarbazine (Matulane®); quinacrine (Atabrine®); rasburicase (Elitek®); rituximab (Rituxan®); ridaforolimus; sargramostim (Leukine®); sargramostim (Prokine®); sorafenib (Nexavar®); streptozocin (Zanosar®); sunitinib maleate (Suten t®); talc (Sclerosol®); tamoxifen (Nolvadex®); temozolomide (Temodar®); teniposide, VM-26 (Vumon®); testolactone (Teslac®); thioguanine, 6-TG (Thioguanine®); thiotepa (Thioplex®); topotecan (Hycamtin®); toremifene (Fareston®); tositumomab (Bexxar®); tositumomab / I-131 tositumomab (Bexxar®); trastuzumab (Herceptin®); tretinoin, ATRA (Vesanoid®); uracil mustard (Uracil Mustard Capsules®); valrubicin (Valstar®); vinblastine (Velban®); vincristine (Oncovin®); vinorelbine (Navelbine®); olaparib (Lynparza®); vorinostat (Zolinza®), and zoledronate (Zometa®), or a pharmaceutically acceptable salt thereof.

[0229] Accordingly, the scope of the present disclosure encompasses the use of an ADC of this disclosure in combination with a second compound selected from estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic / cytostatic agents, antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, angiogenesis inhibitors, PPAR-γ agonists, PPAR-δ agonists, inhibitors of intrinsic multidrug resistance, antiemetic agents, agents useful for the treatment of anemia, agents useful for the treatment of neutropenia, immune enhancing agents, inhibitors of cell proliferation and survival signaling, bisphosphonates, aromatase inhibitors, siRNA therapeutics, γ-secretase and / or NOTCH inhibitors, agents that interfere with receptor tyrosine kinases (RTKs), agents that interfere with cell cycle checkpoints, and any of the therapeutic agents listed above.

[0230] Yet another example of the present disclosure is a method of treating cancer comprising administering a therapeutically effective amount of an ADC of the present disclosure in combination with paclitaxel or trastuzumab.

[0231] The therapeutic combinations disclosed herein may be used in combination with one or more other active agents, including, but not limited to, other anti-cancer agents used to prevent, treat, control, ameliorate, or reduce the risk of a particular disease or condition (e.g., a cell proliferation disorder). In one embodiment, an ADC of the present disclosure is combined with one or more other anti-cancer agents for use in preventing, treating, controlling, ameliorating, or reducing the risk of a particular disease or condition for which an ADC of the present disclosure is useful. Such other active agents may be administered before, simultaneously with, or sequentially with an ADC of the present disclosure by a route and in an amount commonly used.

[0232] The present disclosure also includes pharmaceutical compositions useful for treating or preventing cancer, comprising a therapeutically effective amount of an ADC of the present disclosure and a second compound selected from an estrogen receptor modulator, an androgen receptor modulator, a retinoid receptor modulator, a cytotoxic / cytostatic agent, an antiproliferative agent, a prenyl-protein transferase inhibitor, an HMG-CoA reductase inhibitor, an HIV protease inhibitor, a reverse transcriptase inhibitor, an angiogenesis inhibitor, a PPAR-γ agonist, a PPAR-δ agonist, an inhibitor of cell proliferation and survival signaling, a bisphosphonate, an aromatase inhibitor, an siRNA therapeutic, a γ-secretase and / or NOTCH inhibitor, an agent that interferes with a receptor tyrosine kinase (RTK), an agent that interferes with a cell cycle checkpoint, and any of the therapeutic agents listed above.

[0233] The present disclosure further relates to a method of treating cancer in a human patient, comprising administering to the patient a PD-1 antagonist. The compound of the disclosure and the PD-1 antagonist may be administered simultaneously or sequentially.

[0234] In certain embodiments, the PD-1 antagonist is an anti-PD-1 antibody or an antigen-binding fragment thereof. In alternative embodiments, the PD-1 antagonist is an anti-PD-L1 antibody or an antigen-binding fragment thereof. In some embodiments, the PD-1 antagonist is an anti-PD-1 antibody independently selected from pembrolizumab, nivolumab, cemiplimab, sintilimab, tislelizumab, atezolizumab (MPDL3280A), camrelizumab, and toripalimab. In other embodiments, the PD-L1 antagonist is an anti-PD-L1 antibody independently selected from atezolizumab, durvalumab, and avelumab.

[0235] In one embodiment, the PD-1 antagonist is pembrolizumab. In a particular subembodiment, the method comprises administering 200 mg of pembrolizumab to the patient about every three weeks. In another subembodiment, the method comprises administering 400 mg of pembrolizumab to the patient about every six weeks.

[0236] In a further subembodiment, the method comprises administering 2 mg / kg of pembrolizumab to the patient about every 3 weeks. In a particular subembodiment, the patient is a pediatric patient.

[0237] In some embodiments, the PD-1 antagonist is nivolumab. In certain subembodiments, the method comprises administering 240 mg of nivolumab to the patient about every two weeks. In other subembodiments, the method comprises administering 480 mg of nivolumab to the patient about every four weeks.

[0238] In some embodiments, the PD-1 antagonist is cemiplimab. In certain embodiments, the method comprises administering 350 mg of cemiplimab to the patient about every three weeks.

[0239] In some embodiments, the PD-1 antagonist is atezolizumab. In particular subembodiments, the method comprises administering 1200 mg of atezolizumab to the patient about every three weeks.

[0240] In some embodiments, the PD-1 antagonist is durvalumab. In particular subembodiments, the method comprises administering 10 mg / kg of durvalumab to the patient about every two weeks.

[0241] In some embodiments, the PD-1 antagonist is avelumab. In certain subembodiments, the method comprises administering 800 mg of avelumab to the patient about every two weeks.

[0242] When an ADC of the present disclosure is administered in combination with an anti-human PD-1 antibody (or antigen-binding fragment thereof), the anti-human PD-1 antibody (or antigen-binding fragment thereof) can be administered simultaneously with, before, or after the ADC of the present disclosure. The anti-human PD-1 antibody (or antigen-binding fragment thereof) and / or the ADC of the present disclosure or a pharmaceutically acceptable salt thereof can be administered separately, by the same or different administration routes, or together with one or more other agents in the same pharmaceutical composition. The weight ratio of the anti-human PD-1 antibody (or antigen-binding fragment thereof) to the ADC of the present disclosure can vary and depends on the therapeutically effective dose of each agent. Generally, a therapeutically effective dose of each is used. Combinations comprising at least one anti-human PD-1 antibody (or antigen-binding fragment thereof), an ADC of the present disclosure, and optionally other active agents generally contain a therapeutically effective dose of each active agent. In such combinations, the anti-human PD-1 antibody (or antigen-binding fragment thereof), the ADC of the present disclosure, and other active agents can be administered separately or together. In addition, the administration of one element may be prior to, concurrent to, or subsequent to the administration of other agent(s).

[0243] In one embodiment, the disclosure provides an anti-human PD-1 antibody (or antigen-binding fragment thereof), and / or a compound of Formula IV, and at least one other active agent as a combined preparation for simultaneous, separate, or sequential use in the treatment of cancer.

[0244] The present disclosure also provides for the use of an ADC of the present disclosure to treat cancer, wherein the patient has previously (e.g., within 24 hours) been treated with an anti-human PD-1 antibody (or antigen-binding fragment thereof).The present disclosure also provides for the use of an anti-human PD-1 antibody (or antigen-binding fragment thereof) to treat a cell proliferative disorder, wherein the patient has previously (e.g., within 24 hours) been treated with an antibody-linker-payload compound (antibody drug conjugate), an ADC of the present disclosure.

[0245] The present disclosure further relates to a method of treating cancer, comprising administering to a subject in need thereof a combination therapy comprising (a) an ADC of the present disclosure and (b) an anti-human PD-1 antibody (or antigen-binding fragment thereof), wherein the anti-human PD-1 antibody (or antigen-binding fragment thereof) is administered once every 21 days.

[0246] Furthermore, the present disclosure relates to a method of treating cancer, the method comprising administering to a subject in need thereof a combination therapy comprising (a) an ADC of the present disclosure and (b) an anti-human PD-1 antibody (or antigen-binding fragment thereof). In certain embodiments, the cancer arises as one or more solid tumors or lymphomas. In further particular embodiments, the cancer is selected from the group consisting of advanced or metastatic solid tumors and lymphomas. In even further particular embodiments, the cancer is selected from the group consisting of malignant melanoma, head and neck squamous cell carcinoma, MSI-H cancer, MMR-deficient cancer, non-small cell lung cancer, urothelial carcinoma, gastric or gastroesophageal junction adenocarcinoma, breast adenocarcinoma, and lymphoma. In additional embodiments, the lymphoma is selected from the group consisting of diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, small lymphocytic lymphoma, mediastinal large B-cell lymphoma, splenic marginal zone B-cell lymphoma, extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (malt), nodal marginal zone B-cell lymphoma, lymphoplasmacytic lymphoma, primary effusion lymphoma, Burkitt's lymphoma, anaplastic large cell lymphoma (primary cutaneous type), anaplastic large cell lymphoma (systemic type), peripheral T-cell lymphoma, angioimmunoblastic T-cell lymphoma, adult T-cell lymphoma / leukemia, extranodal nasal NK / T-cell lymphoma, enteropathic T-cell lymphoma, gamma / delta hepatosplenic T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, mycosis fungoides, and Hodgkin's lymphoma. In certain embodiments, the cell proliferative disorder is metastatic cancer, for example, liver metastasis from colon cancer. In additional embodiments, the cell proliferative disorder is cancer classified as stage III cancer or stage IV cancer. In exemplary of these embodiments, the cancer is not surgically resectable.

[0247] In embodiments of the methods disclosed herein, the anti-human PD-1 antibody (or antigen-binding fragment thereof) is administered by intravenous infusion or subcutaneous injection.

[0248] In one embodiment, the disclosure provides a composition comprising an ADC of the disclosure, a pharmaceutically acceptable carrier, and an anti-human PD-1 antibody (or antigen-binding fragment thereof).

[0249] In another embodiment, the disclosure provides a composition comprising a compound of formula IV, a pharmaceutically acceptable carrier, and pembrolizumab.

[0250] In one embodiment, the disclosure provides a composition comprising a compound of Formula IV, a pharmaceutically acceptable carrier, and two additional therapeutic agents, one of which is an anti-human PD-1 antibody (or antigen-binding fragment thereof) and the other of which is an anti-cancer agent.

[0251] The ADCs of the present disclosure can be used in conjunction with antiemetics to treat nausea or vomiting, including acute, delayed, late, and anticipatory emesis, which may result from the use of the ADCs of the present disclosure alone or in conjunction with radiation therapy. To prevent or treat emesis, the ADCs of the present disclosure can be used in conjunction with other antiemetics, particularly neurokinin-1 receptor antagonists, 5HT3 receptor antagonists such as ondansetron, granisetron, tropisetron, and zatisetron, GABAB receptor agonists such as baclofen, corticosteroids such as Decadron (dexamethasone), Kenalog, Aristocort, Nasalide, Preferid, Benecorten, or other antiemetics, such as those described in U.S. Pat. Nos. 2,789,118, 2,999,999, and 3,000,000. and 3,749,712. Other antidopaminergic agents, such as phenothiazines (e.g., prochlorperazine, fluphenazine, thioridazine, and mesoridazine), metoclopramide, aprepitant, fosaprepitant, or dronabinol, as disclosed in U.S. Pat. Nos. 0,401, 3,048,581, 3,126,375, 3,929,768, 3,996,359, 3,928,326, and 3,749,712, may be used in combination with anti-dopaminergic agents, such as phenothiazines (e.g., prochlorperazine, fluphenazine, thioridazine, and mesoridazine), metoclopramide, aprepitant, fosaprepitant, or dronabinol. In another example, combination therapy with an antiemetic selected from a neurokinin-1 receptor antagonist, a 5HT3 receptor antagonist, and a corticosteroid is disclosed for treating or preventing emesis that may occur upon administration of an ADC of the present disclosure.

[0252] The ADCs of the disclosure can also be administered with agents useful in the treatment of anemia, such as sustained erythropoiesis receptor activators (e.g., epoetin alfa).

[0253] The ADCs of the present disclosure may also be administered together with an agent useful for treating neutropenia. Such neutropenia therapeutic agents are hematopoietic growth factors that regulate the production and function of neutrophils, such as human granulocyte colony-stimulating factor (G-CSF). An example of G-CSF is filgrastim.

[0254] The ADC of the present disclosure can be useful when co-administered with other treatment modalities, including but not limited to radiation therapy, surgery, and gene therapy.Therefore, in one embodiment, the method for treating cancer described herein can optionally include the administration of an effective amount of radiation therapy, unless otherwise specified.In the case of radiation therapy, gamma rays are preferred.

[0255] The methods of treating cancer described herein may optionally include the administration of an effective amount of radiation (i.e., the methods of treating cancer described herein may optionally include the administration of radiation therapy).

[0256] Methods of treating cancer described herein include methods of treating cancer comprising administering a therapeutically effective amount of an ADC of the present disclosure in combination with radiation therapy and / or in combination with a second compound selected from an estrogen receptor modulator, an androgen receptor modulator, a retinoid receptor modulator, a cytotoxic / cytostatic agent, an antiproliferative agent, a prenyl-protein transferase inhibitor, an HMG-CoA reductase inhibitor, an HIV protease inhibitor, a reverse transcriptase inhibitor, an angiogenesis inhibitor, a PPAR-γ agonist, a PPAR-δ agonist, an inhibitor of intrinsic multidrug resistance, an antiemetic agent, an agent useful for treating anemia, an agent useful for treating neutropenia, an immune enhancing agent, an inhibitor of cell proliferation and survival signaling, a bisphosphonate, an aromatase inhibitor, an siRNA therapeutic, a γ-secretase and / or NOTCH inhibitor, an agent that interferes with a receptor tyrosine kinase (RTK), an agent that interferes with a cell cycle checkpoint, and any of the additional therapeutic agents listed herein.

[0257] Additional embodiments of the present disclosure include the pharmaceutical compositions, combinations, uses, and methods set forth above, and it should be understood that each embodiment may be combined with one or more other embodiments, so long as such combinations are consistent with the description of the embodiment. It should further be understood that the embodiments provided above are understood to include all embodiments, including such embodiments as a result of combinations of embodiments.

[0258] kit In one aspect, a kit is provided that comprises a therapeutically effective amount of an ADC of the present disclosure or a pharmaceutically acceptable salt, solvate, or ester of said compound and a pharmaceutically acceptable carrier, vehicle, or diluent.

[0259] In another aspect, a kit is provided that includes an amount of the ADC of the present disclosure and an amount of at least one additional therapeutic agent listed above, wherein the amounts of the two or more active ingredients produce a desired therapeutic effect.In one embodiment, the compound of the present disclosure and one or more additional therapeutic agents are provided in the same container.In one embodiment, the compound of the present disclosure and one or more additional therapeutic agents are provided in separate containers.

[0260] Composition and Administration Certain aspects of the disclosure pertain to pharmaceutical compositions comprising a therapeutically effective amount of an ADC of the disclosure, or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0261] Another embodiment of the present disclosure pertains to a composition comprising an ADC of the present disclosure, or a pharmaceutically acceptable salt thereof, having a DAR that is a decimal number between 0 and 8. In one embodiment, the composition is a pharmaceutical composition and includes one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0262] Another aspect of the present disclosure pertains to pharmaceutical compositions comprising a therapeutically effective amount of an ADC of the present disclosure, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, as described herein, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0263] Another aspect of the present disclosure pertains to an ADC of the present disclosure described herein, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, for use as a drug or drug moiety.

[0264] Another aspect of the present disclosure pertains to an ADC of the present disclosure described herein, or a tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, in the preparation of a medicament for treating or preventing a tumor.

[0265] In another embodiment, compounds of the present disclosure include those identified herein as examples in the tables below, and pharmaceutically acceptable salts thereof.

[0266] In another embodiment, the disclosure relates to a method of manufacturing a medicament for use in a subject, the method comprising combining an ADC of the disclosure, or a pharmaceutically acceptable salt thereof, with a pharmaceutical carrier or diluent.

[0267] The ADCs of the disclosure can be administered by oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, ICV, intracisternal injection or infusion, subcutaneous injection, or implant), inhalation spray, nasal, vaginal, rectal, sublingual, buccal, or topical routes of administration, and can be formulated, alone or together, into suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants, and vehicles appropriate for each route of administration. The compounds of the disclosure are effective for use in humans, in addition to treating warm-blooded animals.

[0268] The pharmaceutical composition for administering the compound of the present disclosure can be conveniently provided in dosage unit form, and can be prepared by any method well known in the field of pharmacy.All methods include the step of associating active ingredient with carrier, which constitutes one or more accessory ingredients.Generally, pharmaceutical composition is prepared by uniformly and intimately associating active ingredient with liquid carrier or finely divided solid carrier or both, and then, if necessary, shaping the product into desired formulation.In pharmaceutical composition, active compound is contained in an amount sufficient to produce the desired effect on disease process or condition.

[0269] Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, such as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, solutions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient mixed with non-toxic pharmaceutically acceptable excipients suitable for tablet manufacture. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents such as cornstarch or alginic acid; binders such as starch, gelatin, or acacia; and lubricants such as magnesium stearate, stearic acid, or talc. Tablets may be uncoated or may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained effect over a longer period of time.For example, time-delay materials such as glyceryl monostearate or glyceryl distearate may be used.They may also be coated by the techniques described in U.S. Patent Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotic therapeutic tablets for controlled release.Oral tablets may also be formulated for immediate release, such as fast melt tablets or wafers, fast-dissolving tablets, or fast-dissolving films.

[0270] Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin, or olive oil.

[0271] Aqueous suspensions contain the active material in admixture with excipients suitable for the manufacture of aqueous suspensions, such as suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and gum acacia; dispersing or wetting agents, for example, natural phosphatides, such as lecithin, or condensation products of alkylene oxides with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long-chain aliphatic alcohols, for example, heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, for example, polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example, polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0272] Oily suspension can be prepared by suspending active ingredient in vegetable oil, such as peanut oil, olive oil, sesame oil or coconut oil, or mineral oil such as liquid paraffin.Oily suspension can contain thickening agent, such as beeswax, solid paraffin or acetyl alcohol.Sweetener, such as the sweetener described above, and flavoring agent can be added to provide a pleasant oral preparation.These compositions can be preserved by adding antioxidants such as ascorbic acid.

[0273] Dispersible powders and granules suitable for preparation of an aqueous suspension by adding water provide the active ingredient in admixture with a dispersing or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, such as sweeteners, flavorings, and coloring agents, may also be present.

[0274] The pharmaceutical composition of the present disclosure may be in the form of an oil-in-water emulsion.The oily phase can be vegetable oil, such as olive oil or peanut oil, or mineral oil, such as liquid paraffin, or their mixture.Suitable emulsifiers can be naturally occurring gums, such as acacia gum or tragacanth gum, naturally occurring phosphatides, such as soybean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and the condensation products of the partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate.Emulsions can also contain sweeteners and flavoring agents.

[0275] Syrups and elixirs may be formulated with sweetening agents, for example, glycerol, propylene glycol, sorbitol, or sucrose. Such formulations may also contain a demulcent, a preservative, and flavoring and coloring agents.

[0276] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension. This suspension may be formulated according to known techniques using the appropriate dispersing or wetting agent and suspending agent mentioned above. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile fixed oils are conventionally used as solvents or suspending media. For this purpose, any non-irritating fixed oil may be used, including synthetic monoglycerides or diglycerides. Additionally, fatty acids such as oleic acid are used in the preparation of injectables.

[0277] The ADCs of the present disclosure can also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore melts in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycol.

[0278] For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the ADCs of the disclosure are used. Similarly, transdermal patches may be used for topical administration.

[0279] The pharmaceutical compositions and methods of the present disclosure may further comprise other therapeutically active compounds as described herein that are typically applied in the treatment of the above-mentioned pathological conditions.

[0280] For the treatment, prevention, control, amelioration, or reduction of risk of the conditions disclosed herein, suitable dosage levels of the compounds of the present disclosure will generally be about 0.01 to 500 mg / kg of patient body weight / day, which can be administered in single or multiple doses. Suitable dosage levels may be about 0.01 to 250 mg / kg / day, about 0.05 to 100 mg / kg / day, or about 0.1 to 50 mg / kg / day. Within this range, dosages may be 0.05 to 0.5, 0.5 to 5, or 5 to 50 mg / kg / day. For oral administration, the composition may be provided in the form of tablets containing 1.0 to 1000 milligrams of active ingredient, particularly 1.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of active ingredient, for symptomatic adjustment of dosage to the patient being treated. The compound may be administered on a regimen of 1 to 4 times per day, or once or twice daily.

[0281] However, the specific dose level and frequency of administration for a particular patient may vary and will depend upon a variety of factors including the activity of the specific compound used, the metabolic stability and length of action of that compound, age, body weight, general health, sex, diet, mode and time of administration, excretion rate, drug combination, severity of the particular condition, and the host being treated.

[0282] Methods for preparing the compounds of this disclosure are illustrated in the following schemes and examples. Starting materials are made according to procedures known in the art or as illustrated herein.

[0283] Preparation Examples The compounds of the present disclosure can be prepared according to the following schemes and specific examples or modifications thereof using readily available starting materials, reagents and conventional synthesis procedures. Variations not specifically mentioned, which are themselves known to those skilled in the art, can also be used. The general procedures for making the compounds claimed in this disclosure can be readily understood by those skilled in the art from viewing the following schemes and descriptions.

[0284] General Experiment Information: All reactions were magnetically stirred unless otherwise noted. Unless otherwise noted, all reagents and solvents were purchased from commercial sources and used as received. Reaction progress and analysis of synthetic intermediates were assessed by LCMS (with UV detection by ESI, APCI, or other mass detection) using MeCN / water gradients containing either TFA, formic acid, or NH4HCO3 modifiers, where applicable. Silica gel and reversed-phase flash column chromatography were performed using commercially available prepacked columns. Reverse-phase preparative HPLC purifications were performed on a preparative HPLC system with UV and MS detection using MeCN / water gradients containing either TFA, formic acid, or NH4OH modifiers. 1 H NMR spectra were collected at room temperature, chemical shifts are reported in ppm relative to the residual protein-solvent signal, and multiplicities, coupling constants (where applicable), and signal integrals are listed in parentheses. Unless otherwise noted, all ECs presented in the tables 50 Data refer to the cytotoxicity assay described in the biological assay section.

[0285] Synthetic Schemes, Intermediates, and Examples The compounds of the present disclosure can be prepared by methods known in the art of organic synthesis, as illustrated in part by the following general synthetic schemes and specific preparative examples. Starting materials are commercially available or can be prepared by known methods.

[0286] Preparation of intermediate compounds Preparation of intermediate compound i [ka] Step A - Synthesis of compound ii To a stirred mixture of (9H-fluoren-9-yl)methylhydrazinecarboxylate (i, 3.00 g, 11.8 mmol) in DMF (50.0 mL) was added bis(2,5-dioxopyrrolidin-1-yl)carbonate (3.02 g, 11.8 mmol), and the resulting reaction was allowed to stir at 25 °C for 1 hour. The reaction was monitored using TLC (DCM:CH3OD = 10:1). The reaction mixture containing 1-((9H-fluoren-9-yl)methyl)2-(2,5-dioxopyrrolidin-1-yl)hydrazine-1,2-dicarboxylate (ii, 4.66 g, 11.8 mmol) in DMF was used without further purification.

[0287] Step B - Synthesis of compound iii To a stirred mixture of ii (4.66 g, 11.8 mmol) in DMF (50.0 ml), tert-butyl (2-aminoethyl)carbamate (1.89 g, 11.8 mmol) was added, and the resulting reaction was allowed to stir at 25 °C for 16 hours. Water (50.0 mL) was added, and the mixture was extracted with ethyl acetate (2 × 40.0 mL). The combined organic extracts were washed with brine (50.0 mL), dried over NaSO, filtered, and concentrated in vacuo. The resulting residue was purified using flash silica gel chromatography (5% MeOH / DCM) to afford (9H-fluoren-9-yl)methyl 11,11-dimethyl-4,9-dioxo-10-oxa-2,3,5,8-tetraazadodecanoate (iii, 3.7 g, 8.40 mmol) as a solid. LCMS: MS (ESI) m / z: 441.3 [M+H]+ .

[0288] Step C - Synthesis of compound iv To a stirred mixture of iii (3.0 g, 6.8 mmol) in DMF (10 mL) was added piperidine (1.0 mL, 10 mmol), and the resulting reaction was allowed to stir at 25 °C for 30 min. The reaction mixture was monitored using TLC (DCM:CH3OD = 10:1). The reaction mixture was concentrated in vacuo, and the resulting residue was purified using silica gel column flash chromatography (eluted with 0-10% MeOH / DCM) to afford tert-butyl (2-(hydrazinecarboxamido)ethyl)carbamate iv (1.10 g, 5.04 mmol) as an oil. 1 H NMR (500 MHz, CD3OD) δ = 3.28-3.19 (m, 2H), 3.18-3.11 (m, 2H), 1.50-1.36 (m, 9H). No exchangeable protons reported.

[0289] Preparation of intermediate compound vi [ka] Step A - Synthesis of compound vi To a solution of tert-butyl (4-hydroxybenzyl)carbamate (v, 300 mg, 1.344 mmol) in DCM (10 mL) at 0° C. was added DIEA (0.235 mL, 1.34 mmol) and trichloromethyl chloroformate (0.380 g, 1.92 mmol). The resulting reaction was allowed to stir at 0° C. for 30 minutes and at 20° C. for 2 hours. The reaction mixture was concentrated in vacuo to provide 4-(((tert-butoxycarbonyl)amino)methyl)phenyl carbonochloridate vi (300 mg, 1.050 mmol), which was used without further purification.

[0290] Preparation of intermediate compound x [ka] Step A - Synthesis of compound viii To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-proline (vii, 3.4 g, 10 mmol) and HATU (3.83 g, 10.1 mmol) in DMF (30 mL) was added a solution of tert-butyl(2-aminoethyl)carbamate (1.61 g, 10.1 mmol) and DIEA (5.28 mL, 30.2 mmol) at 0° C. The mixture was allowed to stir at 0° C. for 4 hours and then at 25° C. for an additional 17 hours. The crude (9H-fluoren-9-yl)methyl(S)-2-((2-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)pyrrolidine-1-carboxylate (viii, 5 g, 5.73 mmol) was used directly without further purification.

[0291] Step B - Synthesis of compound ix To a solution of viii (4.0 g, 8.3 mmol) in DMF (10 mL) was added piperidine (1.0 mL, 8.3 mmol), and the resulting mixture was allowed to stir for 1 h at 20° C. The mixture was concentrated in vacuo, diluted with DCM (10 mL), filtered, and lyophilized to afford crude tert-butyl (S)-(2-(pyrrolidine-2-carboxamido)ethyl)carbamate (ix, 2.4 g, 6.5 mmol) as an oil, which was used without further purification.

[0292] Step C - Synthesis of Compound x To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)glycylglycine (2.70 g, 7.62 mmol) and HATU (4.35 g, 11.4 mmol) in DMF (10 mL) was added ix (1.96 g, 7.62 mmol), followed by DIEA (3.99 mL, 22.9 mmol). The resulting reaction was allowed to stir at 0° C. for 4 h and then at room temperature for 17 h, then concentrated in vacuo to provide compound x, which was used without further purification. LCMS (ESI) m / z: 616.3 [M+Na+].

[0293] Preparation of intermediate compound xiv [ka] Step A - Synthesis of compound xiii To a stirred mixture of benzyl azetidin-3-ylcarbamate (xi, 500 mg, 2.42 mmol) in DCM (20 mL) was added triethylamine (736 mg, 7.27 mmol), sodium triacetoxyborohydride (771 mg, 3.64 mmol), and tert-butyl 3-formylazetidine-1-carboxylate (xii, 539 mg, 2.91 mmol) at room temperature under an argon atmosphere. The resulting mixture was allowed to stir for 16 h. The reaction mixture was concentrated in vacuo, and the resulting residue was purified using flash silica gel column chromatography (0–100% EtOAc / petroleum ether) to afford tert-butyl 3-((3-(((benzyloxy)carbonyl)amino)azetidin-1-yl)methyl)azetidine-1-carboxylate xiii as an oil. MS: m / z = 376.25 [M+H] + .

[0294] Step B - Synthesis of compound xiv To a stirred mixture of xiii (490 mg, 1.31 mmol) in MeOH (10 mL) was added 10% Pd / C (139 mg, 0.131 mmol) at room temperature under a hydrogen atmosphere. The resulting mixture was allowed to stir for 1 hour, and then the reaction mixture was filtered and washed with MeOH (20 mL). The filtrate was concentrated in vacuo to give tert-butyl 3-((3-aminoazetidin-1-yl)methyl)azetidine-1-carboxylate xiv as a solid. MS: m / z=242.25 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ 3.83(s,2H)3.46-3.42(m,4H),2.59-2.56(m,2H),1.84(s,4H),1.36(s,9H). Preparation of intermediate compound xviii [ka] Step A - Synthesis of compound xvi To a solution of (S)-2-amino-2,3-dimethylbutanoic acid (xv, 400 mg, 3.05 mmol) in THF (10 mL) was added a solution of potassium carbonate (843 mg, 6.10 mmol) in water (5 mL), followed by (9H-fluoren-9-yl)methyl(2,5-dioxopyrrolidin-1-yl)carbonate (1.54 g, 4.57 mmol). The resulting reaction was allowed to stir at 20° C. for 17 hours, and then the reaction mixture was concentrated in vacuo and acidified to pH=2 to form a suspension, which was then filtered to obtain compound (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-2,3-dimethylbutanoic acid xvi as a solid, which was used without further purification. LCMS (ESI) m / z: 354.3 [M+H]+.

[0295] Step B - Synthesis of compound xvii To a solution of xvi (800 mg, 1.36 mmol) in DMF (5 mL) was added HATU (775 mg, 2.04 mmol), followed by tert-butyl (2-(methylamino)ethyl)carbamate (237 mg, 1.36 mmol) and DIEA (0.712 mL, 4.07 mmol). The reaction mixture was allowed to stir at 20 °C for 1 h and then concentrated in vacuo. The resulting residue was purified using silica gel column chromatography (eluting with 0-50% EtOAc / petroleum ether) to afford (9H-fluoren-9-yl)methyl (S)-(1-((2-((tert-butoxycarbonyl)amino)ethyl)(methyl)amino)-2,3-dimethyl-1-oxobutan-2-yl)carbamate xvii as a solid. LCMS (ESI) m / z: 532.3 [M+Na] + .

[0296] Step C - Synthesis of compound xviii To a solution of xvii (400 mg, 0.785 mmol) in DMF (5 mL) was added piperidine (0.50 mL, 0.79 mmol), and the reaction mixture was allowed to stir at 20° C. for 1 hour. The crude mixture was concentrated in vacuo and purified using HPLC (Boston Green ODS 150×30 mm×5 μm, eluted with 15%-35% MeCN / water (containing 0.5% TFA as a modifier)) to give compound tert-butyl (S)-(2-(2-amino-N,2,3-trimethylbutanamido)ethyl)carbamate xviii as a solid. LCMS (ESI) m / z: 288.2 [M+H] + .

[0297] Preparation of intermediate compound xxiv [ka] Step A - Synthesis of Compound xx To a stirred mixture of 3-aminopyrrolidin-2-one (xix, 50.0 mg, 0.499 mmol) and BocO (0.174 mL, 0.749 mmol) in MeOH (2 mL), TEA (0.209 mL, 1.50 mmol) was added, and the resulting reaction was allowed to stir at 70 °C for 16 h. The reaction was monitored using TLC (DCM:CHOD = 10:1). The solvent was removed in vacuo, and the resulting residue was purified using flash silica gel chromatography (0-100% EtOAc / petroleum ether) to give tert-butyl (2-oxopyrrolidin-3-yl)carbamate xx. MS (ESI) m / z: 201.3 [M+H] + Step B - Synthesis of compound xxi To a stirred suspension of NaH (24 mg, 0.60 mmol) in THF (1 mL) was added xx (100 mg, 0.499 mmol) at 0 °C, and the resulting reaction was allowed to stir at 25 °C for 30 min under a N atmosphere. A solution of 2-bromoacetonitrile (71.9 mg, 0.599 mmol) and TBAI (36.9 mg, 0.100 mmol) in THF (1 mL) was added, and the resulting reaction was allowed to stir at 25 °C for 3 h. The reaction was monitored using TLC (EtOAc). Water (30 mL) was added, and the resulting solution was extracted with ethyl acetate (100 mL × 2). The combined organic extracts were washed with brine (100 mL × 2), dried over Na SO , filtered, and concentrated in vacuo. The resulting residue was purified using flash silica gel chromatography (0-100% EtOAc / petroleum) to afford tert-butyl (1-(cyanomethyl)-2-oxopyrrolidin-3-yl)carbamate xxi.

[0298] Step C - Synthesis of compound xxii To a stirred mixture of xxi (1.3 g, 5.4 mmol) in EtOH (15 mL) was added platinum(IV) oxide (0.247 g, 1.09 mmol), and the resulting reaction was allowed to stir at 25 °C under an atmosphere of H for 16 h. The reaction was monitored using TLC (EtOAc). The mixture was filtered, and the filter cake was washed with ethanol (10 mL). The filtrate was concentrated in vacuo, and the resulting residue was purified using preparative HPLC (Welch Xtimate C18 150 × 25 mm × 5 um, 8–100% ACN:water (with TFA as a modifier)), and the solution was lyophilized to give tert-butyl (1-(2-aminoethyl)-2-oxopyrrolidin-3-yl)carbamate xxii. MS (ESI) m / z: 244.2 [M+H] + .

[0299] Step D - Synthesis of compound xxiii To a solution of xxii (400 mg, 1.64 mmol) in DCM (5 mL) was added TFA (1.0 mL, 13 mmol) and the resulting reaction was allowed to stir at 0° C. for 1 hour. The reaction mixture was filtered, concentrated in vacuo, and the resulting residue was lyophilized to give 3-amino-1-(2-aminoethyl)pyrrolidin-2-one xxiii. LCMS (ESI) m / z: 144.1 [M+H] + .

[0300] Step E - Synthesis of compound xxiv To a solution of xxiii (200 mg, 1.40 mmol) in DCM (10 mL) was added TEA (0.389 mL, 2.79 mmol) followed by BocO (0.259 mL, 1.12 mmol) in DCM (10 mL) at 0° C., and the mixture was then allowed to stir at 0° C. for 4 hours. The crude mixture was concentrated in vacuo to give crude compound tert-butyl (2-(3-amino-2-oxopyrrolidin-1-yl)ethyl)carbamate xxiv, which was used in the next step without further purification. LCMS (ESI) m / z: 244.4 [M+H] + .

[0301] Preparation of intermediate compound xxvii [ka] Step A - Synthesis of compound xxvi To a solution of (S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamido)-5-ureidopentanoic acid (xxv, 500 mg, 1.01 mmol) in DMF (5 mL) was added tert-butyl (4-aminobenzyl)carbamate (224 mg, 1.01 mmol), TCFH (339 mg, 1.21 mmol), and 1-methylimidazole (248 mg, 3.02 mmol). The resulting reaction was allowed to stir at 20° C. for 18 hours, and then the reaction mixture was concentrated in vacuo. The resulting residue was purified using preparative HPLC (Boston Green ODS 150 x 30 mm x 5 um, eluting with 48% to 78% acetonitrile / water (containing 0.1% TFA as a modifier)) to give (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-((4-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (xxvi). LCMS (ESI) m / z: 701.4 [M+H] + .

[0302] Step B - Synthesis of compound xxvii To a solution of (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-((4-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (xxvi, 260 mg, 0.371 mmol) in DMF (4 mL) was added piperidine (0.4 mL, 0.4 mmol). The resulting reaction was allowed to stir at 20° C. for 1 hour, then the reaction mixture was concentrated in vacuo to provide compound xxvii, which was used without purification. LCMS (ESI) m / z: 479.3 [M+H + ].

[0303] Preparation of intermediate compound xxxi [ka] Step A - Synthesis of compound xxix To a solution of ((benzyloxy)carbonyl)-L-valyl-L-alanine (xxviii, 1.0 g, 3.1 mmol) in DMF (5 mL) was added HATU (1.30 g, 3.41 mmol). The mixture was stirred at room temperature for 5 minutes, and tert-butyl (4-aminobenzyl)carbamate (0.690 g, 3.10 mmol) was added. After stirring at room temperature for an additional 5 minutes, DIEA (1.63 mL, 9.31 mmol) was added, and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was purified using preparative HPLC (Boston Uni C18 150 x 40 mm x 5 um, eluting with 42% to 72% acetonitrile / water (with 0.1% TFA as a modifier)) to give benzyl ((S)-1-(((S)-1-((4-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (xxix). LCMS (ESI) m / z: 527.3 [M+H] + .

[0304] Step B - Synthesis of Compound xxx To a stirred mixture of xxix (550 mg, 1.044 mmol) in trifluoroethanol (15 mL) was added Pd—C (300 mg, 10% Pd), and the resulting reaction was allowed to stir under a hydrogen atmosphere at 25° C. for 18 hours. The mixture was concentrated in vacuo to give tert-butyl (4-((S)-2-((S)-2-amino-3-methylbutanamido)propanamido)benzyl)carbamate (xxx). LCMS (ESI) m / z: 393.4 [M+H] + Step C - Synthesis of compound xxxi To a solution of 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoic acid (164 mg, 0.968 mmol) in DMF (4 mL) was added HATU (405 mg, 1.07 mmol). After stirring for 5 minutes, xxx (380 mg, 0.968 mmol) and DIEA (0.507 mL, 2.90 mmol) were added, and the resulting reaction was allowed to stir at 20° C. for 2 hours. The resulting residue was purified using preparative HPLC (Boston Green ODS 150 x 30 mm x 5 um, eluted with 30% to 60% acetonitrile / water (containing 0.1% TFA as a modifier)) to give tert-butyl (4-((S)-2-((S)-2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)-3-methylbutanamido)propanamido)benzyl)carbamate (xxxi). LCMS (ESI) m / z: 544.3 [M+H] + .

[0305] The following intermediate compounds of the present disclosure were prepared using the methods described above and substituting the appropriate reactants and / or reagents: [Table 8] Preparation of intermediate compound xli [ka] Step A - Synthesis of compound xxxiv Chromic anhydride (19.7 g, 197 mmol) was added portionwise over 2 hours to a solution of 2-chloro-6-methylnicotinonitrile (xxxiii, 10 g, 66 mmol) in sulfuric acid (70.0 mL, 1310 mmol) while maintaining the reaction mixture at 0° C. After stirring at room temperature for 18 hours, the mixture was poured into ice water and the resulting solid was collected by filtration to give crude 6-chloro-5-(aminocarbonyl)-2-picolinic acid xxxiv.

[0306] Step B - Synthesis of Compound xxxv The mixture of xxxiv was dissolved in POCl3 (50 mL, 536 mmol), and the reaction mixture was heated to 115 °C and allowed to stir at this temperature for 30 minutes. The mixture was then concentrated in vacuo, and the resulting residue was diluted with ice water. The resulting solution was filtered, and the collected solid was dried to give 6-chloro-5-cyanopicolinic acid xxxv as a solid. LC / MS: MS(ESI) m / z: 183.2 [M+H] + .

[0307] Step C - Synthesis of compound xxxvi To a stirred mixture of xxxv (5.60 g, 30.7 mmol) in DMF (50 mL) was added sodium methanethiol (2.15 g, 30.7 mmol). The mixture was allowed to stir at 20 °C for 15 h. The mixture was concentrated in vacuo and purified using flash silica gel column chromatography (0-50% MeOH / DCM) to give 5-cyano-6-(methylthio)picolinic acid xxxvi as a solid. LCMS (ESI) m / z: 195.2 [M+H] + .

[0308] Step D - Synthesis of compound xxxvii To a stirred mixture of xxxvi (200 mg, 1.03 mmol) in THF (5 mL) was added HATU (470 mg, 1.24 mmol), DIPEA (0.540 mL, 3.09 mmol), and tert-butyl 3-aminopropanoate (150 mg, 1.03 mmol). The mixture was allowed to stir at 20 °C for 15 h. The mixture was concentrated in vacuo and purified using flash silica gel column chromatography (0-100% petroleum ether / EtOAc) to afford tert-butyl 3-(5-cyano-6-(methylthio)picolinamido)propanoate xxxvii as an oil. LCMS (ESI): 266.2 [M-56+H] + .

[0309] Step E - Synthesis of compound xxxviii To a stirred mixture of xxxvii (218 mg, 0.678 mmol) in DCM (5 mL) was added m-CPBA (468 mg, 2.71 mmol). The mixture was allowed to stir at 20 °C for 18 h. The mixture was concentrated in vacuo and purified using flash silica gel column chromatography (0-100% petroleum ether / EtOAc) to afford tert-butyl 3-(5-cyano-6-(methylsulfonyl)picolinamido)propanoate xxxviii as a solid. LCMS (ESI): 298.2 [M-56+H] + .

[0310] Step F - Synthesis of compound xxxix A solution of xxxviii (1.2 g, 3.4 mmol) in DCM (18 mL) and TFA (6 mL) was allowed to stir at room temperature for 1 h. The reaction mixture was concentrated in vacuo to give 3-(5-cyano-6-(methylsulfonyl)picolinamido)propanoic acid xxxix as a solid, which was used without further purification. LCMS (ESI) m / z: 298.2 [M+H] + .

[0311] Step G - Synthesis of compound xl To a stirred mixture of xxxix (300 mg, 1 mmol) in DCM (6 mL) was added EDC (232 mg, 1.21 mmol) and N-hydroxysuccinamide (151 mg, 1.31 mmol). The mixture was allowed to stir at 20° C. for 5 hours. The solution was concentrated to give 2,5-dioxopyrrolidin-1-yl 3-(5-cyano-6-(methylsulfonyl)picolinamido)propanoate xl as a solid, which was used without further purification. LCMS (ESI) m / z: 395.0 [M+H] + .

[0312] Step H - Synthesis of compounds xli To a solution of xl (400 mg, 1 mmol) in DMF (10 mL) was added L-alanyl-L-alanine (179 mg, 1.12 mmol) and TEA (0.566 mL, 4.06 mmol). The resulting reaction was allowed to stir at 25 °C for 2 hours, and then the reaction mixture was concentrated in vacuo to give (methyl (3-(5-cyano-6-(methylsulfonyl)picolinamido)propanoyl)-L-alanyl-L-alaninate xli as a solid, which was used without purification. LCMS (ESI) m / z: 440.0 [M+H] + .

[0313] Preparation of intermediate compound xlviii [ka] Step A - Synthesis of compound xliii A solution of 5-bromo-6-hydroxypyridine-3-carboxylic acid (xlii, 600 g, 2.59 mol) and cuprous cyanide (231 g, 2.59 mol) in DMF (3.6 L) was stirred at 125° C. for 12 hours under a N atmosphere. The reaction mixture was cooled to room temperature and quenched with water (5.0 L). The resulting mixture was filtered, and the filter cake was dried under reduced pressure to give 5-cyano-6-hydroxypyridine-3-carboxylic acid (xlii). LCMS: (ESI, m / z): [M−H] - =163 Step B - Synthesis of compound xliv A solution of xliii (600 g, 3.37 mol) and phosphorus oxychloride (1.8 L) was allowed to stir at 110 °C for 1 hour. The reaction was monitored using LCMS and then concentrated in vacuo. The crude product was quenched with water pre-cooled to 10 °C, and the mixture was extracted with EtOAc (3 x 500 mL). The organic layer was washed with brine (500 mL) and dried over sodium sulfate. The mixture was filtered and concentrated in vacuo, and the resulting residue was purified using reverse-phase flash chromatography (silica gel, 25% to 100% petroleum ether / EtOAc) to give 6-chloro-5-cyanopyridine-3-carboxylic acid (xliv). LCMS: (ESI, m / z): [MH] - =181 Step C - Synthesis of compound xlv To a 500 mL three-necked bottle, dimethylformamide (115.5 mL) and xliv (7.7 g, 42 mmol) were added at 25 °C. The reaction was cooled to 10 °C, and (methylsulfanyl)sodium (7.39 g, 105 mmol) was added in several portions. The resulting reaction was allowed to stir at 25 °C for 8 hours, and then the reaction mixture was slowly transferred to HO (1200 mL) and then extracted with ethyl acetate (1 × 700 mL). The pH value of the aqueous layer was adjusted to 2-3 with 1 M HCl, and the resulting solution was filtered to give 5-cyano-6-(methylsulfanyl)pyridine-3-carboxylic acid (xliv) as a solid. LCMS: (ESI, m / z): [MH] - =193 Step D - Synthesis of compound xlvi To a solution of xlv (7.4 g, 38 mmol) in DCM (185 mL), m-CPBA (26.3 g, 152 mmol) was added, and the resulting reaction was stirred at 45° C. under a nitrogen atmosphere for 24 hours. The reaction was quenched with saturated sodium bisulfite at 0° C. and concentrated in vacuo. To the resulting residue, 2-methyltetrahydrofuran was added, and the mixture was filtered and concentrated in vacuo. The resulting residue was purified using silica gel column chromatography (eluting with DCM / MeOH) to give compound xlv. LCMS: (ESI, m / z): [M+H] + =227.05.

[0314] Step E - Synthesis of compound xlvii Intermediate xlvi (0.57 g, 2.5 mmol) and HATU (1.0 g, 2.7 mmol) were dissolved in 10 mL of DMF, and the resulting solution was stirred at 25 °C for 30 minutes. tert-Butyl 3-(5-cyano-6-(methylsulfonyl)nicotinamido)propanoate (0.65 g, 1.8 mmol) was added, and the resulting reaction was cooled to 10 °C. DIPEA (0.873 mL, 5.00 mmol) was added dropwise to the reaction mixture at 10 °C, and the reaction was allowed to stir at 25 °C for 2 hours. It was then diluted with water and extracted with EtOAc (3 × 10 mL). The combined organic extracts were concentrated in vacuo, and the resulting residue was purified using silica gel column chromatography (eluting with 2:1 to 1:1 hexanes:EtOAc) to give compound xlvii. 1 H NMR(500MHz,CD3OD)δ 8.49(d,J=2.0Hz,1H),8.07(d,J=2.0Hz,1H),4.12(s,3H),2.92(t,J=6.9Hz,2H),1.88(t,J=6.9Hz,2H),0.74(s,9H). Step F - Synthesis of compound xlviii Compound xlvii (80 g, 0.226 mmol) was dissolved in 1,4-dioxane (160 mL), and to the resulting mixture was added 4 M HCl solution in 1,4-dioxane (480 mL). The resulting reaction was allowed to stir at 25° C. for 12 hours, and then the reaction mixture was filtered and concentrated in vacuo to provide compound xlviii, which was used without further purification. 1 H NMR(500MHz,CD3OD)δ 9.22(d,J=1.9Hz,1H),8.80(d,J=1.9Hz,1H),3.66(t,J=5.8Hz,2H),3.44(s,3H),2.67(t,J=6.8Hz,2H). Preparation of intermediate compound l [ka] Step A - Synthesis of compound xlix To a stirred mixture of 3-(5-cyano-6-(methylsulfonyl)picolinamido)propanoic acid (xxxix, 1 g, 3 mmol) in THF (5 mL) was added HATU (1.54 g, 4.04 mmol), DIPEA (1.76 mL, 10.1 mmol), and tert-butyl glycylglycinate (0.633 g, 3.36 mmol). The reaction was allowed to stir at 20 °C for 15 h. The reaction mixture was then concentrated in vacuo, and the resulting residue was purified using flash silica gel column chromatography (0-100% petroleum ether / EtOAc) to afford tert-butyl (3-(5-cyano-6-(methylsulfonyl)picolinamido)propanoyl)glycylglycinate xlix as a solid. LCMS (ESI): 468.2 [M+H] + Step B - Synthesis of Compound l A solution of xlix (1.2 g, 2.6 mmol) in DCM (18 mL) and TFA (6 mL) was stirred at room temperature for 1 hour. The reaction mixture was concentrated in vacuo to give (3-(5-cyano-6-(methylsulfonyl)picolinamido)propanoyl)glycylglycine l as a solid, which was used without further purification. LCMS (ESI) m / z: 412.1 [M+H] + .

[0315] Preparation of intermediate compound liv [ka] Step A - Synthesis of compound lii To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanyl-L-alanine (1.5 g, 3.9 mmol) in DMF (30 mL) was added HATU (1.49 g, 3.92 mmol) and the resulting solution was allowed to stir at room temperature for 10 minutes. tert-Butyl 3-(aminomethyl)azetidine-1-carboxylate (0.80 g, 4.3 mmol) was added and the reaction was allowed to stir for an additional 10 minutes. DIPEA (2.06 mL, 11.8 mmol) was added and the reaction was allowed to stir at 20° C. for an additional 3 hours. The reaction mixture was concentrated in vacuo, and the resulting residue was purified using preparative HPLC (Boston Uni C18 40 x 150 x 5 um, eluting with 40% to 70% acetonitrile in acetonitrile / water (with 0.1% TFA as a modifier)) to give tert-butyl 3-((5S,8S)-1-(9H-fluoren-9-yl)-5,8-dimethyl-3,6,9-trioxo-2-oxa-4,7,10-triazaundecan-11-yl)azetidine-1-carboxylate (lii). LCMS (ESI) m / z: 551.2 [M+H] + .

[0316] Step B - Synthesis of Compound liii To a solution of lii (2.37 g, 4.30 mmol) in DMF (15 mL) was added piperidine (1.5 mL, 15.15 mmol), and the resulting reaction was allowed to stir at 25° C. for 3 hours. The reaction mixture was dried using lyophilization to provide tert-butyl 3-(((S)-2-((S)-2-aminopropanamido)propanamido)methyl)azetidine-1-carboxylate (liii), which was used without further purification. LCMS (ESI): 329.2 [M+H]+.

[0317] Step C - Synthesis of compound liv To a solution of 3-(5-cyano-6-(methylsulfonyl)picolinamido)propanoic acid (xxxix, 400 mg, 1.35 mmol) in DMF (1.5 mL) was added HATU (767 mg, 2.02 mmol) and DIPEA (0.470 mL, 2.69 mmol), and the reaction was allowed to stir at room temperature for 10 minutes. Compound liii (803 mg, 1.35 mmol) (55%) was added, and the reaction was allowed to stir at 20° C. for an additional 30 minutes. The reaction mixture was concentrated in vacuo, and the resulting residue was purified using preparative HPLC (Boston Uni C18 40 x 150 x 5 um, eluting with 15% to 45% acetonitrile in acetonitrile / water (with 0.1% TFA as a modifier)) to give tert-butyl 3-((4S,7S)-13-(5-cyano-6-(methylsulfonyl)pyridin-2-yl)-4,7-dimethyl-3,6,9,13-tetraoxo-2,5,8,12-tetraazatridecyl)azetidine-1-carboxylate (liv). LCMS (ESI): 608.2 [M+H]+.

[0318] The following intermediate compounds of the present disclosure were prepared using the methods described above and substituting the appropriate reactants and / or reagents: [Table 9] Preparation of intermediate compound lxv [ka] Step A - Synthesis of compound lxiii To a solution of (3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl)-L-alanyl-L-alanine (lxii, 700 mg, 2.25 mmol) in DCM (10 mL) and MeOH (10 mL) was added (4-aminophenyl)methanol (332 mg, 2.70 mmol), followed by EEDQ (834 mg, 3.37 mmol). The resulting reaction was cooled to 20° C. and allowed to stir at this temperature for 18 hours. The reaction mixture was concentrated in vacuo and the resulting residue was purified using silica gel chromatography (0–10% MeOH in DCM) to afford 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)amino-1-oxopropan-2-yl)propanamide (lxiii). LCMS (ESI) m / z: 439.3 [M+Na]+.

[0319] Step B - Synthesis of compound lxiv To a solution of lxiii (200 mg, 0.48 mmol) in DMF (5 mL) was added bis(4-nitrophenyl)carbonate (161 mg, 0.528 mmol) and DIEA (0.109 mL, 0.624 mmol), and the resulting reaction was cooled to 20 °C and allowed to stir at this temperature for 18 h. The reaction mixture was diluted with water (30 mL), and the resulting solution was concentrated using lyophilization to give 4-((S)-2-((S)-2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)propanamido)benzyl(4-nitrophenyl)carbonate (lxiv). LCMS (ESI) m / z: 582.3 [M+H] + .

[0320] Step C - Synthesis of compound lxv To a solution of lxiv (200 mg, 0.344 mmol) in DMF (5 mL) was added tert-butyl 3-(aminomethyl)azetidine-1-carboxylate (64.1 mg, 0.344 mmol), and the resulting reaction was allowed to stir at 20° C. for 1 hour. The reaction mixture was directly purified using preparative HPLC (Boston Green ODS 150×30 mm×5 μm, eluted with 40%-60% MeCN / water (containing 0.01% TFA as a modifier)) to provide tert-butyl 3-(((((4-((S)-2-((S)-2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)propanamido)propanamido)benzyl)oxy)carbonyl)amino)methyl)azetidine-1-carboxylate (lxv). LCMS(ESI)m / z:529.3[M+H + -Boc].

[0321] Preparation of intermediate compound lxxi [ka] Step A - Synthesis of compound lxvii To a solution of (4-aminophenyl)methanol (lxvi, 6.44 g, 52.3 mmol) in DCM / MeOH (2:1) (60 mL) was added a stirred mixture of EEDQ (4.85 g, 19.6 mmol) and (((9H-fluoren-9-yl)methoxy)carbonyl)-L-alanyl-L-alanine (5.0 g, 13 mmol) at room temperature. The resulting reaction was allowed to stir at room temperature for 18 hours, and then the reaction mixture was concentrated in vacuo. The resulting residue was suspended in TBME (80 mL) and stirred for 30 minutes, then the mixture was filtered and the filter cake was washed with TBME (20 mL). The combined filtrate and washings were concentrated in vacuo to give (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-((4-(hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)carbamate (lxvii).

[0322] Step B - Synthesis of compound lxviii DIPEA (0.143 mL, 0.820 mmol) was added to a stirred mixture of bis(4-nitrophenyl)carbonate (0.150 g, 0.492 mmol) and lxvii (0.2 g, 0.410 mmol) in DMF (2 mL) at room temperature, and the resulting reaction was allowed to stir at room temperature for 2 hours. The reaction mixture was poured into HO (40 mL), and the resulting mixture was filtered. The filter cake was washed with water (10 mL) and then dried under vacuum with toluene to give (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-((4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)carbamate (lxviii). LCMS(ESI)m / z:675.3[M+H] + .

[0323] Step C - Synthesis of compound lxix A solution of lxviii (2 g, 3.06 mmol), tert-butyl (2-(hydrazinecarboxamido)ethyl)carbamate (0.803 g, 3.68 mmol), and HOBt (0.141 g, 0.919 mmol) in DMF (20 mL) was stirred at room temperature for 5 minutes. Pyridine (0.496 mL, 6.13 mmol) was added, and the resulting reaction was stirred at room temperature for 18 hours. The reaction mixture was then concentrated in vacuo. The resulting residue was purified using silica gel column flash chromatography (DCM / MeOH 0-30%) to give 4-((S)-2-((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)propanamido)benzyl 11,11-dimethyl-4,9-dioxo-10-oxa-2,3,5,8-tetraazadodecanoate (lxix). LCMS(ESI)m / z:732.4[M+H] + .

[0324] Step D - Synthesis of compounds lxx Diethylamine (0.50 mL, 4.9 mmol) was added to a stirred mixture of lxix (1.1 g, 1.5 mmol) in DMF (10 mL) at room temperature, and the resulting reaction was allowed to stir for 2 hours. The reaction mixture was then concentrated in vacuo to give 4-((S)-2-((S)-2-aminopropanamido)propanamido)benzyl 11,11-dimethyl-4,9-dioxo-10-oxa-2,3,5,8-tetraazadodecanoate (lxx), which was used without further purification.

[0325] Step E - Synthesis of compound lxxi DIPEA (0.679 mL, 3.89 mmol) was added to a stirred mixture of 2,5-dioxopyrrolidin-1-yl 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoate (414 mg, 1.55 mmol) and lxx (660 mg, 1.30 mmol) in DMF (8 mL) at room temperature, and the resulting reaction was allowed to stir for 2 hours. The reaction mixture was directly purified using preparative HPLC (Boston Uni C18 50 x 40 mm x 5 um, eluting with 19%-49% acetonitrile / water (with 0.1% TFA as a modifier)) to give 4-((S)-2-((S)-2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)propanamido)benzyl 11,11-dimethyl-4,9-dioxo-10-oxa-2,3,5,8-tetraazadodecanoate (lxxi). LCMS (ESI) m / z: 661.3 [M+H] + .

[0326] Preparation of intermediate compound lxxv [ka] Step A - Synthesis of compound lxxiii To a solution of (3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl)glycylglycylglycine (1xxii, 0.50 g, 1.5 mmol) in DMF (5 mL) was added HATU (0.670 g, 1.76 mmol). The resulting solution was allowed to stir at room temperature for 10 minutes, and then a solution of tert-butyl methyl (2-(methylamino)ethyl)carbamate (0.304 g, 1.616 mmol) in DMF (0.1 mL) was added, and the resulting reaction was allowed to stir at room temperature for 10 minutes. DIEA (0.770 mL, 4.41 mmol) was added, and the reaction was allowed to stir at room temperature for 1 hour. The reaction mixture was directly purified using reverse-phase HPLC (Boston Green ODS 150 x 30 mm x 5 um, 23% to 53% acetonitrile / water (with 0.1% TFA as a modifier)) and concentrated in vacuo to give tert-butyl (15-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-methyl-4,7,10,13-tetraoxo-3,6,9,12-tetraazapentadecyl)(methyl)carbamate (lxxiii, 0.5 g, 0.979 mmol).

[0327] Step B - Synthesis of compound lxxiv Compound lxxiii (200 mg, 0.392 mmol) was dissolved in a 3:1 mixture of DCM:TFA (4 mL), and the resulting solution was allowed to stir for 1 hour at 20° C. The reaction mixture was concentrated in vacuo to provide 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-(5-methyl-6,9,12-trioxo-2,5,8,11-tetraazatridecan-13-yl)propanamide (lxxiv), which was used without further purification.

[0328] Step C - Synthesis of compound lxxv To a solution of lxxiv (100 mg, 0.244 mmol) in DCM (10 mL) was added DIEA (0.032 mL, 0.184 mmol) and 4-(((tert-butoxycarbonyl)amino)methyl)phenyl carbonochloridate (35 mg, 0.12 mmol). The resulting reaction was cooled to 0° C. and allowed to stir at this temperature for 30 minutes. The reaction mixture was allowed to warm to room temperature and then stirred for an additional hour. The reaction mixture was concentrated in vacuo, and the resulting residue was purified using reverse-phase HPLC (Boston Green ODS 150 x 30 mm x 5 um, 23% to 53% acetonitrile / water (with 0.1% TFA as a modifier)) and concentrated to give 4-(((tert-butoxycarbonyl)amino)methyl)phenyl (15-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-3-methyl-4,7,10,13-tetraoxo-3,6,9,12-tetraazapentadecyl)(methyl)carbamate (lxxv). LCMS (ESI) m / z: 660.3 [M+H] + .

[0329] Preparation of intermediate compound lxxxiv [ka] Step A - Synthesis of compound lxxvii To a solution of lxxvi (10 g, 28 mmol) in THF (100 mL) and toluene (40 mL) was added lead tetraacetate (17.5 g, 39.5 mmol), and the resulting reaction was heated to 85 °C and allowed to stir at this temperature for 18 h. The reaction mixture was concentrated in vacuo, and the resulting residue was purified using silica gel flash chromatography (EtOAc / petroleum ether 30-50%) to afford lxxvii (11.2 g, 21.3 mmol) as a solid. LCMS (ESI) m / z: 391.2 [M+Na] + .

[0330] Step B - Synthesis of compound lxxviii To a solution of lxxvii (2.5 g, 6.8 mmol) in DCM (30 mL) was added TFA (1.05 mL, 13.6 mmol) and tert-butyl 2-hydroxyacetate (4.48 g, 33.9 mmol). The resulting reaction was allowed to stir at 25 °C for 18 h. The reaction mixture was then diluted with saturated aqueous NaHCO (10 mL) and extracted with DCM (3 × 30 mL). The combined organic extracts were dried over sodium sulfate, filtered, and concentrated in vacuo, and the resulting residue was purified using silica gel column chromatography (EtOAc / petroleum ether 5-9%) to afford lxxviii (1.02 g, 1.852 mmol) as a solid. LCMS (ESI) m / z: 463.1 [M+Na] + .

[0331] Step C - Synthesis of compound lxxix To a solution of lxxviii (1.5 g, 3.4 mmol) in DCM (9 mL) was added TFA (3.0 mL, 39 mmol), and the resulting reaction was allowed to stir at room temperature for 1 hour. The reaction mixture was concentrated in vacuo to give lxxix (1.2 g, 2.2 mmol) as an oil, which was used without further purification. LCMS (ESI) m / z: 407.1 [M+H + ].

[0332] Step D - Synthesis of compounds lxxx To a solution of lxxix (1.0 g, 2.6 mmol) in DMF (10 mL) was added HATU (1.19 g, 3.12 mmol), and the resulting solution was allowed to stir at room temperature for 5 minutes. tert-Butyl (3-aminobicyclo[1.1.1]pentan-1-yl)carbamate (0.671 g, 3.38 mmol) and DIEA (1.36 mL, 7.80 mmol) were added, and the reaction was allowed to stir at 20 °C for 2 hours. The reaction mixture was concentrated in vacuo, and the resulting residue was purified using preparative HPLC (Boston Uni C18 150 mm x 40 mm x 5 μm, eluting with acetonitrile / water (0.1% TFA) at a flow rate of 60 mL / min, eluting at 37% to 67%) to give lxxx (680 mg, 1.148 mmol) as a solid. LCMS (ESI) m / z: 565.4 [M+H]+ .

[0333] Step E - Synthesis of compounds lxxxi To a solution of lxxx (370 mg, 0.655 mmol) in DMF (4 mL) was added piperidine (0.40 mL, 0.66 mmol). The resulting reaction was allowed to stir at 20° C. for 1 hour, and then the reaction mixture was concentrated in vacuo to give lxxxi (360 mg, 0.526 mmol) as a solid, which was used without further purification. LCMS (ESI) m / z: 343.2 [M+H] + .

[0334] Step F - Synthesis of compound lxxxii To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-phenylalanine (407 mg, 1.05 mmol) in DMF (5 mL) was added HATU (300 mg, 0.789 mmol), and the resulting solution was allowed to stir at room temperature for 5 minutes. Compound lxxvi (360 mg, 0.526 mmol) and DIEA (0.275 mL, 1.58 mmol) were added, and the resulting reaction was allowed to stir at 20 °C for 1 hour. The reaction mixture was directly purified using preparative HPLC (Boston Uni C18 150 mm x 40 mm x 5 μm, 45%-75% MeCN / water (0.1% TFA), flow rate 60 mL / min) to afford lxxxii (180 mg, 0.253 mmol) as a solid. LCMS (ESI) m / z: 712.3 [M+H] + .

[0335] Step F - Synthesis of compound lxxxiii To a solution of lxxxii (180 mg, 0.253 mmol) in DMF (2 mL) was added piperidine (0.20 mL, 0.25 mmol). The resulting reaction was allowed to stir at 20° C. for 30 minutes, and then the reaction mixture was concentrated in vacuo to give lxxxiii (150 mg, 0.184 mmol) as a solid, which was used without further purification. LCMS (ESI) m / z: 490.2 [M+H] + .

[0336] Step G - Synthesis of compound lxxxiv To a solution of (3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl)glycylglycine (226 mg, 0.797 mmol) in DMF (2 mL) was added HATU (202 mg, 0.531 mmol), and the resulting reaction was allowed to stir at room temperature for 5 minutes. Compound lxxxiii (130 mg, 0.266 mmol) and DIEA (0.139 mL, 0.797 mmol) were added, and the resulting reaction was allowed to stir at 20°C for 2 hours. The reaction mixture was then concentrated in vacuo, and the resulting residue was purified using preparative HPLC (Boston Prime C18 150 mm x 40 mm x 5 μm, eluting with MeCN / water (0.1% TFA) 20% to 40% at a flow rate of 25 mL / min) to afford lxxxiv (80 mg, 0.079 mmol) as a solid. LCMS(ESI)m / z:755.3[M+H] + .

[0337] Preparation of intermediate compound lxxxvii [ka] Step A - Synthesis of compounds lxxxvi To a solution of lxxxv (420 mg, 1.02 mmol) in DMF (7 mL) was added HATU (466 mg, 1.23 mmol), and the resulting solution was allowed to stir at room temperature for 5 minutes. tert-Butyl (3-aminobicyclo[1.1.1]pentan-1-yl)carbamate (202 mg, 1.02 mmol) and DIEA (0.535 mL, 3.06 mmol) were added, and the resulting reaction was allowed to stir at 20° C. for 2 hours. The reaction mixture was concentrated in vacuo to give lxxxvi (600 mg, 0.608 mmol) as a solid, which was used without further purification. LCMS (ESI) m / z: 592.3 [M+H] + .

[0338] Step B - Synthesis of Compound lxxxvii To a solution of lxxxvi (600 mg, 1.01 mmol) in DMF (6 mL) was added piperidine (0.60 mL, 1.0 mmol), and the resulting reaction was allowed to stir at 20° C. for 1 hour. The reaction mixture was concentrated in vacuo, and the resulting residue was purified using preparative HPLC (Boston Prime C18 150 mm x 40 mm x 5 um, eluting with MeCN / water (0.1% TFA) 5% to 35% at a flow rate of 25 mL / min) to afford lxxxvii (400 mg, 0.974 mmol) as a solid. LCMS (ESI) m / z: 370.3 [M+H] + .

[0339] Preparation of intermediate compound xciii [ka] Step A - Synthesis of Compound lxxxix To a solution of lxxxviii (3.5 g, 17 mmol) in AcOH (30 mL) was added furan-2,5-dione (1.68 g, 17.1 mmol). The resulting reaction was allowed to stir at 20° C. for 18 hours, and then the reaction mixture was concentrated in vacuo to give lxxxix (5 g, 16.50 mmol) as a solid, which was used without further purification. LCMS (ESI) m / z: 203.2 [M+H-Boc] + .

[0340] Step B - Synthesis of compound xc To a solution of lxxxix (5.0 g, 16 mmol) in toluene (40 mL) was added dimethylacetamide (2 mL) and triethylamine (6.92 mL, 49.6 mmol). The resulting reaction was heated to 110° C. and allowed to stir at this temperature for 18 hours. The reaction mixture was then cooled to room temperature and concentrated in vacuo. The resulting residue was purified using preparative HPLC (Boston Green ODS 150 mm x 30 mm x 5 μm, eluting with MeCN / water (0.1% TFA) 20%-40% at a flow rate of 25 mL / min) to afford xc (3.08 g, 7.32 mmol) as a solid. LCMS (ESI) m / z: 229.1 [M+H-56] + .

[0341] Step C - Synthesis of Compound xci To a solution of xc (3.08 g, 10.8 mmol) in DCM (15 mL) was added TFA (5.0 mL, 65 mmol) and the resulting reaction was allowed to stir at 25° C. for 3 h. The reaction mixture was concentrated in vacuo to give xci (2.4 g) as an oil, which was used without further purification. LCMS (ESI) m / z: 185.2 [M+H] + .

[0342] Step D - Synthesis of compound xcii To a solution of xci (0.542 g, 2.94 mmol) in DMF (10 mL) was added 2,5-dioxopyrrolidin-1-yl 2,5,8,11,14,17,20,23-octaoxahexacosane-26-oate (1.5 g, 2.9 mmol), followed by NMM (0.596 g, 5.89 mmol), and the resulting reaction was allowed to stir at 20 °C for 18 h. The reaction mixture was directly purified using preparative HPLC (Boston Uni C18 150 mm x 40 mm x 5 um, flow rate 60 mL / min, eluting with MeCN / water (0.1% TFA) 5%-35%) to afford xcii (730 mg, 1.26 mmol) as an oil. LCMS (ESI) m / z: 579.3 [M+H] + .

[0343] Step E - Synthesis of compound xciii To a solution of xcii (0.20 g, 0.35 mmol) and lxxxvii (0.153 g, 0.415 mmol) in DMF (2 mL) was added NMM (0.105 g, 1.04 mmol) at 0 °C, followed by slow addition of HATU (0.131 g, 0.346 mmol). The resulting reaction was allowed to stir at 0 °C for 3 hours, and then the reaction mixture was concentrated in vacuo. The resulting residue was purified using preparative HPLC (Boston Prime C18 150 mm x 40 mm x 5 μm, eluting with MeCN / water (0.1% TFA) 15% to 45% at a flow rate of 25 mL / min) to afford xciii (130 mg, 0.119 mmol) as a powder. LCMS (ESI) m / z: 930.7 [M+H] + .

[0344] Preparation of intermediate compound xcviii [ka] Step A - Synthesis of compound xciv To a solution of ((benzyloxy)carbonyl)glycylglycine (1.0 g, 3.8 mmol) and N-hydroxysuccinimide (0.52 g, 4.5 mmol) in dioxane (7 mL) was slowly added a solution of DCC (0.930 g, 4.51 mmol) in anhydrous dioxane (7 mL). The reaction mixture was allowed to stir at 20° C. for 12 hours, and then the reaction mixture was concentrated in vacuo to give xciv (1.3 g, 2.147 mmol) as a solid, which was used without further purification. LCMS (ESI) m / z: 364.2 [M+H] + .

[0345] Step B - Synthesis of compound xcv A stirred mixture of xcv (1.3 g, 3.6 mmol) and sodium bicarbonate (0.361 g, 4.29 mmol) in water (10 mL) was added to a solution of L-phenylalanylglycine (0.954 g, 4.29 mmol) in dioxane (20 mL). The resulting reaction was allowed to stir at 20 °C for 18 hours, and then the reaction mixture was concentrated in vacuo. The resulting residue was purified using preparative HPLC (Boston Uni C18 150 mm x 40 mm x 5 um, eluting with 15%-45% acetonitrile / water (0.1% TFA) at a flow rate of 60 mL / min) to afford xcv (1.3 g, 2.76 mmol) as a solid. LCMS (ESI) m / z: 471.3 [M+H] + .

[0346] Step C - Synthesis of compound xcvi To a solution of xcv (500 mg, 1.06 mmol) in DMF (7 mL) was added HATU (445 mg, 1.17 mmol), and the resulting solution was allowed to stir at room temperature for 5 minutes. tert-Butyl (3-aminobicyclo[1.1.1]pentan-1-yl)carbamate (211 mg, 1.06 mmol) and Hunig's base (0.557 mL, 3.19 mmol) were added, and the resulting reaction was allowed to stir at 20 °C for 2 hours. The reaction mixture was concentrated in vacuo, and the resulting residue was purified using preparative HPLC (Boston Green ODS 150 mm x 30 mm x 5 μm, eluting with 30%-60% acetonitrile / water (0.1% TFA) at a flow rate of 25 mL / min) to afford xcvi (645 mg, 0.991 mmol) as a solid. LCMS (ESI) m / z: 651.3 [M+H] + .

[0347] Step D - Synthesis of compound xcvii To a mixture of xcvi (645 mg, 0.991 mmol) in trifluoroethanol (15 mL) was added 10% Pd / C (300 mg), and the mixture was stirred under 1 atmosphere of hydrogen at 25° C. for 18 hours. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to give xcvii (544 mg, 0.769 mmol) as an oil. LCMS (ESI) m / z: 517.3 [M+H] + .

[0348] Step E - Synthesis of compound xcviii To a solution of 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoic acid (49.1 mg, 0.290 mmol) in DMF (2 mL) was added HATU (121 mg, 0.319 mmol), and the resulting solution was stirred at room temperature for 5 minutes. Compound xcvii (150 mg, 0.290 mmol) and Hunig's base (0.152 mL, 0.871 mmol) were added, and the resulting reaction was stirred at 20 °C for 2 hours. The reaction mixture was then concentrated in vacuo. The resulting residue was purified using preparative HPLC (Boston Green ODS 150 mm × 30 mm × 5 μm, eluting with 23%–53% acetonitrile / water (0.1% TFA) at a flow rate of 25 mL / min) to afford compound xcviii (70 mg, 0.101 mmol) as a solid. LCMS(ESI)m / z:668.3[M+H] + .

[0349] Preparation of intermediate compound xcx [ka] To a solution of (8S,10S)-6,8,11-trihydroxy-8-(2-hydroxyacetyl)-1-methoxy-10-(((1S,3R,4aS,9S,9aR,10aS)-9-methoxy-1-methyloctahydro-1H-pyrano[4',3':4,5]oxazolo[2,3-c][1,4]oxazin-3-yl)oxy)-7,8,9,10-tetrahydrotetracene-5,12-dione (xcix, 200 mg, 0.312 mmol) in methanol (5 mL) and water (5 mL) was added a solution of sodium periodate (80 mg, 0.37 mmol) in water (1 mL), and the resulting reaction was allowed to stir at room temperature for 30 minutes. The solvent was slowly removed from the reaction mixture in vacuo at 25 °C over 18 hours, and the resulting residue xcix was used without further purification. LCMS(ESI)m / z:628.6[M+H] + .

[0350] Example 1 Preparation of linker-payload 1 [ka] Step A - Synthesis of Compound I-1b To a stirred solution of L-alanyl-L-alanine (I-1a, 1.00 g, 6.24 mmol) and TEA (1.74 mL, 12.5 mmol) in DCM (2 mL) was added 2,5-dioxopyrrolidin-1-yl 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoate (2.09 g, 7.87 mmol) at 25° C. The mixture was allowed to stir at 25° C. for 18 hours. The mixture was concentrated in vacuo and purified using preparative HPLC (YMC-Triart Prep C18 150 x 40 mm x 7 um, eluting with 7% to 37% acetonitrile / water (with 0.1% TFA as a modifier)) to give (3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl)-L-alanyl-L-alanine as solid I-1b. LCMS (ESI) m / z: 312.1 [M+H] + .

[0351] Step B - Synthesis of Compound I-1c To a stirred mixture of I-1b (300 mg, 0.964 mmol) in DMF (4 mL) was added HATU (733 mg, 1.93 mmol), and the resulting reaction was allowed to stir for 10 min at 25° C. Then, tert-butyl (2-aminoethyl)carbamate (154 mg, 0.964 mmol) in DMF (1 mL), and DIPEA (0.505 mL, 2.89 mmol) were added to the above mixture, and the mixture was allowed to stir at 25° C. for 25 min. The solution was purified using preparative HPLC (Boston Uni C18 40 x 150 x 5 um, 8% to 100% acetonitrile / water (with 0.1% TFA as a modifier)) and lyophilized to give tert-butyl (2-((S)-2-((S)-2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)propanamido)propanamido)ethyl)(methyl)carbamate I-1c as an oil. LCMS (ESI) m / z: 490.0 [M+Na] + .

[0352] Step C - Synthesis of Compound I-1d A solution of I-1c (300 mg, 0.642 mmol) and TFA (2 mL, 0.642 mmol) in DCM (6 mL) was allowed to stir at 0° C. for 1 hour. The reaction was monitored using LCMS, and the resulting reaction was concentrated in vacuo at 0° C. to give compound 3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-N-((S)-1-(((S)-1-((2-(methylamino)ethyl)amino)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)propanamide I-1d as an oil, which was used in the next step without further purification. LCMS (ESI) m / z: 368.0 [M+H] + .

[0353] Step D - Synthesis of Compound 1 To a solution of xcx (150 mg, 0.239 mmol) in DMF (2 mL) was added HATU (182 mg, 0.478 mmol). After stirring the reaction mixture for 10 min, I-1d (88 mg, 0.24 mmol) in DMF (0.5 mL) was added and the resulting reaction was stirred for 10 min. DIPEA (0.125 mL, 0.717 mmol) was added and the reaction mixture was stirred for an additional 1 h at 20 °C. The crude mixture was purified by preparative HPLC (YMC-Triart Prep C18 Purification using a 150 x 40 mm x 7 um column, eluting with 25%-55% acetonitrile / water (7 mM ammonium formate as modifier) ​​yielded (2S,4S)-N-(2-((S)-2-((S)-2-(3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamido)propanamido)ethyl)-2,5,12-trihydro- The hydroxy-7-methoxy-4-(((1S,3R,4aS,9S,9aR,10aS)-9-methoxy-1-methyloctahydro-1H-pyrano[4',3':4,5]oxazolo[2,3-c][1,4]oxazin-3-yl)oxy)-N-methyl-6,11-dioxo-1,2,3,4,6,11-hexahydrotetracene-2-carboxamide 1 was obtained as a solid. 1H NMR(400MHz,CDCl3)δ 13.94(br s,1H),13.37(br s,1H),8.03(d,J=7.58Hz,1H),7.78(t,J=8.07Hz,1H),7.39(d,J=8.31Hz,1H),7.00-7.12(m,1H),6.77(br d,J=7.34Hz,1H),6.69(s,2H),6.17-6.33(m,1H),5.53(br s,1H),5.27-5.40(m,1H),4.70(s,1H),4.47(s,1H),4.21-4.44(m,2H),3.99-4.13(m,5H),3.78-3.96(m,3H),3.54- 3.72(m,2H),3.36-3.50(m,8H),2.93-3.11(m,2H),2.67-2.85(m,2H),2.30-2.65(m,4H),1.66-1.84(m,7H),1.38(br d,J=6.36Hz,6H),LCMS(ESI)m / z:977.3[M+H] + . The following exemplary linker-payload compounds of the present disclosure were prepared using the methods described in Example 1 and substituting appropriate intermediates, and other reactants and / or reagents: [Table 10] TIFF2025542248000143.tif196165 TIFF2025542248000144.tif193166 TIFF2025542248000145.tif198165 The following illustrative examples of the present disclosure were prepared using the methods described in Example 1 (Steps B, C, and D) and substituting the appropriate intermediates, reactants, and / or reagents: [Table 11] The following illustrative examples of the present disclosure were prepared using the methods described in Example 1 (Steps C and D) and substituting the appropriate intermediates, reactants and / or reagents: [Table 12] TIFF2025542248000150.tif215165 TIFF2025542248000151.tif201166 TIFF2025542248000152.tif63165 Example 2 Preparation of linker-payload compound 34 [ka] To a solution of compound 19 (80 mg, 0.058 mmol) in DMF (0.3 mL) was added piperidine (0.010 mL, 0.058 mmol), and the resulting reaction was allowed to stir at 0 °C for 10 minutes. The reaction mixture was purified using preparative HPLC (C18-1 150 x 30 mm x 5 um, eluted with 25% to 56% acetonitrile / water (with 8 mM ammonium formate as a modifier)) to give 5-cyano-N-((12S,15S)-12,15-dimethyl-1,11,14,17-tetraoxo-1-((2S,4S)-2,5,12-trihydroxy-7-methoxy-4-(((1S,3R,4aS,9S,9aR,10aS) )-9-Methoxy-1-methyloctahydro-1H-pyrano[4',3':4,5]oxazolo[2,3-c][1,4]oxazin-3-yl)oxy)-6,11-dioxo-1,2,3,4,6,11-hexahydrotetracen-2-yl)-2,6,10,13,16-pentaazanonadecan-19-yl)-6-(methylsulfonyl)picolinamide 34 was obtained as a solid. 1H NMR(400MHz,CD3OD)δ=8.50(dd,J=8.1,10.3Hz,1H),8.39(s,2H),8.29(dd,J=3. 8,8.1Hz,1H),7.88(dd,J=2.1,7.4Hz,1H),7.81-7.73(m,1H),7.50(d,J=8.6Hz,1 H),5.31(q,J=5.0Hz,1H),5.14-5.06(m,1H),4.59(s,1H),4.31(s,1H),4.16-4. 03(m,3H),3.96(s,4H),3.77(td,J=6.1,11.9Hz,1H),3.63-3.55(m,2H),3.47(br d,J=11.7Hz,1H),3.42-3.36(m,5H),3.33(s,3H),3.31-3.25(m,2H),3.10-2.98(m,3H),2.95(br d,J=6.9Hz,4H),2.65(br s,2H),2.57-2.46(m,2H),2.28(br s,2H),1.93-1.77(m,5H),1.77-1.58(m,2H),1.31-1.20(m,9H).LCMS(ESI)m / z:1162.3[M+H] + . The following exemplary linker-payload compounds of the present disclosure were prepared using the methods described in the examples above and substituting the appropriate intermediates, reactants and / or reagents: [Table 13] Example 3 Preparation of linker-payload compound 36 [ka] Step A - Synthesis of Compound I-36b To a solution of (2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)acetamido)methyl acetate (669 mg, 1.82 mmol)) in DCM (10 mL) was added TFA (0.466 mL, 6.05 mmol) and 3-aminobicyclo[1.1.1]pentan-1-ol (I-36a, 120 mg, 1.21 mmol). The resulting reaction was allowed to stir at 25° C. for 18 hours, then the reaction mixture was concentrated in vacuo and the resulting residue was purified using preparative HPLC (Boston Prime C18 150 mm x 40 mm x 5 um, eluting with 15%-45% acetonitrile / water (0.1% TFA) at a flow rate of 25 mL / min) to afford I-36b (230 mg, 0.524 mmol) as a solid. LCMS (ESI) m / z: 408.3 [M+H] + .

[0354] Step B—Synthesis of Compound I-36c To a solution of xcx (200 mg, 0.319 mmol) in DMF (4 mL) was added HATU (145 mg, 0.382 mmol), and the resulting solution was allowed to stir at room temperature for 10 minutes. A solution of compound I-36b (130 mg, 0.319 mmol) in DMF (0.4 mL) was added, and the resulting solution was allowed to stir at room temperature for 10 minutes. Hunig's base (124 mg, 0.956 mmol) was added, and the resulting reaction was allowed to stir at room temperature for 20 minutes. The reaction mixture containing compound I-36c (300 mg, 0.147 mmol) in DMF was used without further purification.

[0355] Step C - Synthesis of Compound I-36d To a solution of I-36c (300 mg, 0.147 mmol) in DMF (4.4 mL) was added piperidine (0.4 mL, 4.04 mmol), and the resulting reaction was allowed to stir at 20 °C for 10 min. The reaction mixture was directly purified using preparative HPLC (Phenomenex Gemini-NX 150 mm × 30 mm × 5 μm; 23% to 53% acetonitrile / water (7 mM HCOONH) at a flow rate of 25 mL / min) to afford I-36d (50 mg, 0.057 mmol) as a solid. LCMS (ESI) m / z: 795.5 [M+H]+ .

[0356] Step D - Synthesis of Compound 36 To a solution of (3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl)glycylglycine (27 mg, 0.094 mmol) in DMF (0.2 mL) was added HATU (47.8 mg, 0.126 mmol), and the resulting solution was allowed to stir at room temperature for 10 min. Compound I-36d (50 mg, 0.063 mmol) was added, and the mixture was stirred at room temperature for 10 min. Hunig's base (0.033 mL, 0.189 mmol) was then added, and the resulting reaction was allowed to stir at 25 °C for 30 min. The reaction mixture was directly purified using preparative HPLC (Phenomenex Gemini-NX 150 mm × 30 mm × 5 μm; 23%–53% acetonitrile / water (7 mM HCOONH) at a flow rate of 25 mL / min) to give 36 (14.7 mg, 0.014 mmol) as a solid. 1 H NMR(400MHz,DMSO-d6)δ=8.49-8.38(m,2H),8.20(br s,1H),8.06(br s,2H),7.86(br s,2H),7.60(br s,1H),6.93(s,2H),5.31(br s,1H),5.19(br s,1H),4.94(br s,1H),4.51(br s,3H),4.16(br s,1H),4.05(br s,1H),3.93(br s,4H),3.71-3.50(m,10H),3.44(m,2H),2.97-2.77(m,2H),2.61(br s,1H),2.35(br s,4H),2.28-2.18(m,3H),2.06(br s,6H),1.60(br s,2H),1.14(m,3H).LCMS(ESI)m / z:1060.6[M+H + ]. Example 4 Antibody Conjugation Protocols for Preparing Antibody Drug Conjugates Examples 37-68 Exemplary linker-payload compounds of the present disclosure were conjugated to anti-TROP2 antibodies using the following conjugation protocol: The antibody (humanized x[TACSTD2_H]mAb (sacituzumab (S375C)) IgG1 / kappa) (20 mg) was used as received in 9% sucrose with Hist (10 mM pH 6.5). The antibody was diluted to approximately 7 mg / ml in 90% buffer / 10% DMSO. A 10 mM solution of linker-payload compound (3.1 equivalents) in DMSO was added, and the resulting conjugation reaction was allowed to stir at room temperature for 2 hours. The reaction mixture was then purified by exchange into 10 mM histidine pH 6.5 using a desalting column on an AKTA chromatography system, followed by the addition of 9% sucrose. Using this methodology, the following antibody-drug conjugates of the present disclosure were produced: [Table 14] TIFF2025542248000157.tif102166 Example 5 Antibody Conjugation Protocol for Preparing Antibody Drug Conjugates Examples 69-70 Exemplary linker-payload compounds of the present disclosure were conjugated to anti-TROP2 antibodies using the following conjugation protocols to yield antibody drug conjugate Examples 69-71, respectively: The antibody (humanized x[TACSTD2_H] mAb (sacituzumab (S375C)) IgG1 / kappa) (20 mg) was exchanged into 40 mM Tris-acetate, 1 mM EDTA, pH 8.3. The antibody was diluted to approximately 10 mg / ml in 90% buffer / 10% DMF. A 10 mM solution of an exemplary linker-payload compound (5.5 equivalents) in DMF was added, and the resulting conjugation reaction was allowed to stir overnight at room temperature. The reaction mixture was then exchanged into 10 mM histidine pH 6.5 using a desalting column on an AKTA chromatography system, followed by purification using the addition of 9% sucrose.

[0357] Using this methodology, the following antibody drug conjugates of the present disclosure were made: [Table 15] Example 6 TROP2 BxPC-3 Cytotoxicity Assay Protocols for Examples 37-41, 43-52, 57-58, and 60-70 Exemplary TROP2 antibody drug conjugates of the present disclosure (Examples 37-41, 43-53, 57-58, 60-68) were subjected to a cell-based cytotoxicity assay (BxPC-3 cells, CellTiter-Glo® 2.0 cytotoxicity assay) utilizing the following protocol: BxPC-3 cells were cultured in a T75 flask in RPMI 1640 medium (Gibco™ 72400-047) supplemented with 10% FB (Gibco™ 26140-079). The cells were washed once with PBS (calcium- and magnesium-free), and then 2 mL of 0.25% trypsin-EDTA (Thermo Fisher™ catalog number 25200056) was added. The flask was incubated at 37°C for approximately 3 minutes. Next, 10 mL of cell culture medium was added, and the cells were dissociated by pipetting up and down several times. The cell mixture was transferred to a 15 mL conical tube and centrifuged at 300 g for 5 minutes. The cell pellet was resuspended in 2 mL of cell culture medium, and the cells were counted using a Vi-CELL. Cells were seeded into 96-well plates (Corning™ Catalog No. 3904) in 90 μl of cell culture medium per well (3,000 cells per well). After 24 hours, serial dilutions of small molecules and antibody-drug conjugates in cell culture medium were prepared, and 10 μL of the dilutions were added to each well (total volume of 100 μL per well). Only the inner 60 wells on the plate were used for drug treatment and untreated controls. The plate was incubated in a cell culture incubator for 96 hours and then equilibrated to room temperature for approximately 30 minutes. CellTiter-Glo™ buffer was thawed and allowed to equilibrate to room temperature. The appropriate volume of CellTiter-Glo™ buffer was transferred to the amber bottle containing CellTiter-Glo™ substrate to reconstitute the lyophilized enzyme / substrate mix (Promega™ Catalog No. G7573). Next, 100 μl of CellTiter-Glo™ reagent was added to each well and the contents were mixed on an orbital shaker for 2 minutes to induce cell lysis. The plate was incubated at room temperature for 10 minutes to stabilize the luminescence signal. Luminescence was recorded on a PerkinElmer Multimode Plate Reader EnVision™. GraphPad Prism 8 [4-parameter nonlinear regression, y = Bottom + (Top - Bottom) / (1 + (IC 50 / x) HillSlope )] was used to analyze the data.

[0358] Exemplary examples of the present disclosure were tested and the results are provided below: [Table 16] Example 7 TROP2 BxPC-3 Cytotoxicity Assay Protocol for Examples 42, 54, and 59 Exemplary TROP2 antibody drug conjugates of the present disclosure (Examples 31, 41, and 46) were subjected to a cell-based cytotoxicity assay (BxPC-3 cells, CellTiter-Glo® 2.0 cytotoxicity assay) utilizing the following protocol: Step 1: Seed a 384-well assay plate on day 0 (45 μL / well) BxPC-3 cells (in a sample vial) were rapidly thawed in a cryovial by incubating in a 37°C water bath for less than 1 minute until only a small amount of ice remained in the sample vial. The vial was removed from the water bath and wiped with 70% ethanol. The cells were transferred from the vial to a sterile centrifuge tube containing 8 mL of prewarmed cell culture medium (RPMI-1640 (catalog no. 30-2001) + 10% FBS + 1% P / S). An additional 1 mL of medium was added to the vial to ensure complete transfer of the cells to the centrifuge tube. The cells were then centrifuged (150 x g) for 5 minutes. The supernatant was aspirated, and the cell pellet was resuspended in 10–20 mL of cell culture medium (RPMI-1640 (catalog no. 30-2001) + 10% FBS + 1% P / S). The cells were counted using Vi-cell and 1500 cells / well were prepared in 45 μL of medium. Then, 45 μL / well of cells were placed into a Corning® 384-well low-flange white flat-bottom polystyrene TC-treated microplate (Corning, Cat. No. 3570) using a Standard Cassette Combi. The plate was spun down in a centrifuge (150×g) for 30 seconds.

[0359] Step 2: Add antibody-drug conjugate on day 1 The centrifuge tubes containing the exemplary antibody-drug conjugates and reference stocks were removed and allowed to thaw at room temperature. The tubes were then centrifuged (2000 × g) for 30 seconds. A 10x intermediate assay plate (Waters plate, catalog number 186002632) was prepared using a Bravo liquid handler, with serial dilutions performed using the appropriate buffer (10 mM pH 6.5 histidine 9% sucrose buffer). Media (without cells) was used for Max_E. 5 μL of the 10x stock from the intermediate plate was then added to the assay plate using a Bravo liquid handler (using a very slow speed to avoid disturbing the cell monolayer). The plate was then spun down in a centrifuge (150 × g) for 30 seconds.

[0360] Step 3: Perform CellTiter-Glo 2.0 assay (Promega, catalog number G9242) (TROP2) on day 5 The CellTiter-Glo™ 2.0 Reagent was thawed overnight at 4°C (during this time, the reagent was not exposed to temperatures above 25°C). The kit was allowed to equilibrate to room temperature for 30 minutes, and then 20 μl of CellTiter-Glo™ 2.0 Reagent was added to the 50 μl of media containing the cells using a Standard Cassette Combi. The contents were mixed on an orbital shaker for 2-3 minutes to induce cell lysis, and the plate was spun down (150 x g) for 30 seconds. The plate was then incubated at room temperature for 5 minutes to stabilize the luminescence signal. Luminescence was recorded and EC was determined using an integration time of 0.25-1 second per well as a guideline. 50 values ​​were calculated.

[0361] Exemplary examples of the present disclosure were tested and the results are provided below: [Table 17]

Claims

1. Compounds having structural formula I: 【Chemistry 1】 and pharmaceutically acceptable salts thereof, wherein R ２ is H or C １ ~C ６ When R is alkyl, １ is -X-Y-Z-R ３ Selected from: R ２ is H or C １ ~C ６ alkyl or R １ and R ２ are joined to form a 4- to 6-membered monocyclic heterocycloalkylene group or a 5- to 11-membered bicyclic spirocyclic heterocycloalkylene group, each of which is bonded to a ring carbon atom by -X-Y-Z-R ３ is substituted with; R ３ teeth, 【Chemistry 2】 Selected from: X is -(CH ２ ) ｎ -N(R ４ ) -, -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-C(O)R ５ -, -(CH ２ ) ｎ -N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ )-, 5- or 6-membered monocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -N(R ４ )-, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -, -(CH ２ ) ｎ -R ５ -N(R ４ )C(O)O-(CH ２ ) ｎ R ５ -N(R ４ ) -, -(CH ２ ) ｎ -NHC(O)O-(CH ２ ) ｎ -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -R ５ -O-C(O)-N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ ) -, -R ５ -NHC(O)CH ２ OCH ２ N (R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )C(O)NH-N(R ４ ) -, -R ５ -N(R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-(CH ２ ) ｎ -N(R ４ )- and -R ５ -O-CH ２ N (R ４ )—selected from; Y is a bond or —C(O)C(R ４ ) (R ６ ) NH—; Z is -C(O)CH(R ７ )NHC(O)CH(R ８ ) NH—; R ４ is H, C １～６ alkyl, or 【Transformation 3】 and R ５ is C ６ ~C １０ arylene, 4- to 6-membered monocyclic heterocycloalkylene, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, C ３ ~C ７ Monocyclic cycloalkylene, C ５ ~C １１ selected from bridged bicyclic cycloalkylene, -NH-NH-, and -NHC(O)NH-, wherein said 4-6 membered monocyclic heterocycloalkylene may optionally be substituted on a ring carbon atom with an oxo group; R ６ , R ７ and R ８ is H, -CH ２ CH ２ CH ２ NHC(O)NH ２ , benzyl, and a naturally occurring amino acid side chain; R ９ Ha-CH ２ NHC(O)-(CH ２ CH ２ O) ｑ -CH ３ and m is 0, 1 or 2; each occurrence of n is independently an integer from 0 to 4; q is selected from 4, 8, 10, and 12).

2. R ３ but 【Chemistry 4】 2. The compound of claim 1, wherein:

3. R ３ but, 【Transformation 5】 2. The compound of claim 1, wherein:

4. 4. The compound of claim 2 or 3, wherein m is 1.

5. R １ Ga-X-Y-Z-R ３ and R ２ is H or C １ ~C ６ 5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R is alkyl.

6. R １ and R ２ are joined to form a 4- to 6-membered monocyclic heterocycloalkylene group or a 5- to 11-membered bicyclic spirocyclic heterocycloalkylene group, each of which is bonded to a ring carbon atom by -X-Y-Z-R ３ 5. The compound of claim 1, 2 or a pharmaceutically acceptable salt thereof, wherein R is substituted with R.

7. R ２ 7. The compound of claim 1, wherein R is H, or a pharmaceutically acceptable salt thereof.

8. 8. The compound of any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein X is a 5- or 6-membered monocyclic heterocycloalkylene or a 5- to 11-membered bicyclic spirocyclic heterocycloalkylene.

9. Xが、-(CH ２ ) ｎ -N(R ４ )-、-R ５ -N(R ４ )-、-(EH ２ ) ｎ -N(R ４ )-C(O)R ５ -、-(EH ２ ) ｎ -N(R ４ )-(C １ ~C ６ アルキレン)-N(R ４ )-、-(EH ２ ) ｎ -R ５ -N(R ４ )-、-(EH ２ ) ｎ -R ５ -、-(EH ２ ) ｎ -R ５ -NHC(O)O-(CH ２ ) ｎ R ５ -N(R ４ )-、-(EH ２ ) ｎ -NHC(O)O-(CH ２ ) ｎ -R ５ -N(R ４ )-、-(EH ２ ) ｎ -R ５ -O-C(O)-N(R ４ )-(C １ ~C ６ アルキレン)-N(R ４ )-、R ５ -NHC(O)H ２ OCH ２ N(R) ４ )C(O)CH ２ N(R) ４ )-、-(EH ２ ) ｎ -N(R ４ )C(O)NH-N(R ４ )-、-R ５ -N(R ４ )C(O)CH ２ N(R) ４ )-、-(EH ２ ) ｎ -N(R ４ )-CH ２ -N(R ４ )- and -R ５ -O-CH ２ N (R ４ )-; R ４ each occurrence is independently H, methyl, or Fmoc; R ５ each occurrence of is -NH-NH-, -NHC(O)NH, 【Transformation 6】 9. The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, independently selected from:

10. 10. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein Y is a bond.

11. Y is -C(O)C(R ４ ) (R ６ ) NH—, and R ６ is H, benzyl, or isopropyl, and R ４ 10. The compound of claim 1, wherein is H or methyl, or a pharmaceutically acceptable salt thereof.

12. Z is -C(O)CH(R ７ )NHC(O)CH(R ８ ) NH—, and R ７ and R ８ is H, methyl, isopropyl, and —CH ２ CH ２ CH ２ NHC(O)NH ２ 12. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, each independently selected from:

13. The -X-Y-Z-R ３ The base is 【Transformation 7】 【change】 【change】 【change】 13. The compound of any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, selected from:

14. A compound, or a pharmaceutically acceptable salt thereof, selected from: Table 1

15. An antibody-drug conjugate having structural formula (XXIII): 【Transformation 8】 or a pharmaceutically acceptable salt thereof, L is an antibody; R ２ is H or C １ ~C ６ is alkyl; R ３’ teeth, 【Chemistry 9】 (Wherein, ** represents R ３’ to L) Selected from: X is -(CH ２ ) ｎ -N(R ４ ) -, -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-C(O)R ５ -, -(CH ２ ) ｎ -N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ )-, 5- or 6-membered monocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -N(R ４ )-, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -, -(CH ２ ) ｎ -R ５ -N(R ４ )C(O)O-(CH ２ ) ｎ R ５ -N(R ４ ) -, -(CH ２ ) ｎ -NHC(O)O-(CH ２ ) ｎ -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -R ５ -O-C(O)-N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ ) -, -R ５ -NHC(O)CH ２ OCH ２ N (R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )C(O)NH-N(R ４ ) -, -R ５ -N(R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-CH ２ -N(R ４ )- and -R ５ -O-CH ２ N (R ４ )—selected from; Y is a bond or —C(O)C(R ４ ) (R ６ ) NH—; Z is -C(O)CH(R ７ )NHC(O)CH(R ８ ) NH—; R ４ is H, C １～６ alkyl, or 【Chemistry 10】 and R ５ is C ６ ~C １０ arylene, 4- to 6-membered monocyclic heterocycloalkylene, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, C ３ ~C ７ Monocyclic cycloalkylene, C ５ ~C １１ selected from bridged bicyclic cycloalkylene, -NH-NH-, and -NHC(O)NH-, wherein said 4-6 membered monocyclic heterocycloalkylene may optionally be substituted on a ring carbon atom with an oxo group; R ６ , R ７ and R ８ is H, -CH ２ CH ２ CH ２ NHC(O)NH ２ , benzyl, and a naturally occurring amino acid side chain; R ９ Ha-CH ２ NHC(O)-(CH ２ CH ２ O) ｑ -CH ３ and m is 0, 1 or 2; each occurrence of n is independently an integer from 0 to 4; q is selected from 4, 8, 10, and 12).

16. An antibody-drug conjugate having structural formula (XXIV): 【Chemistry 11】 or a pharmaceutically acceptable salt thereof, A is a 4- to 6-membered monocyclic heterocycloalkylene or a 5- to 11-membered bicyclic spirocyclic heterocycloalkylene; L is an antibody; R ３’ teeth, 【Chemistry 12】 (Wherein, ** represents R ３’ to L) Selected from: X is -(CH ２ ) ｎ -N(R ４ ) -, -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-C(O)R ５ -, -(CH ２ ) ｎ -N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ )-, 5- or 6-membered monocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -N(R ４ )-, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -, -(CH ２ ) ｎ -R ５ -N(R ４ )C(O)O-(CH ２ ) ｎ R ５ -N(R ４ ) -, -(CH ２ ) ｎ -NHC(O)O-(CH ２ ) ｎ -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -R ５ -O-C(O)-N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ ) -, -R ５ -NHC(O)CH ２ OCH ２ N (R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )C(O)NH-N(R ４ ) -, -R ５ -N(R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-CH ２ -N(R ４ )- and -R ５ -O-CH ２ N (R ４ )—selected from; Y is a bond or —C(O)C(R ４ ) (R ６ ) NH—; Z is -C(O)CH(R ７ )NHC(O)CH(R ８ ) NH—; R ４ is H, C １～６ alkyl, or 【Chemistry 13】 and R ５ is C ６ ~C １０ arylene, 4- to 6-membered monocyclic heterocycloalkylene, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, C ３ ~C ７ Monocyclic cycloalkylene, C ５ ~C １１ selected from bridged bicyclic cycloalkylene, -NH-NH-, and -NHC(O)NH-, wherein said 4-6 membered monocyclic heterocycloalkylene may optionally be substituted on a ring carbon atom with an oxo group; R ６ , R ７ and R ８ is H, -CH ２ CH ２ CH ２ NHC(O)NH ２ , benzyl, and a naturally occurring amino acid side chain; R ９ Ha-CH ２ NHC(O)-(CH ２ CH ２ O) ｑ -CH ３ and m is 0, 1 or 2; each occurrence of n is independently an integer from 0 to 4; q is selected from 4, 8, 10, and 12).

17. 16. The antibody-drug conjugate of claim 15, selected from: Table 2 or a pharmaceutically acceptable salt thereof, X is -(CH ２ ) ｎ -N(R ４ ) -, -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-C(O)R ５ -, -(CH ２ ) ｎ -N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ )-, 5- or 6-membered monocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -N(R ４ )-, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -, -(CH ２ ) ｎ -R ５ -N(R ４ )C(O)O-(CH ２ ) ｎ R ５ -N(R ４ ) -, -(CH ２ ) ｎ -NHC(O)O-(CH ２ ) ｎ -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -R ５ -O-C(O)-N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ ) -, -R ５ -NHC(O)CH ２ OCH ２ N (R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )C(O)NH-N(R ４ ) -, -R ５ -N(R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-CH ２ -N(R ４ )- and -R ５ -O-CH ２ N (R ４ )—selected from; Y is a bond or —C(O)C(R ４ ) (R ６ ) NH—; Z is -C(O)CH(R ７ )NHC(O)CH(R ８ ) NH—; R ４ is H, C １～６ alkyl, or 【Chemistry 14】 and R ５ is C ６ ~C １０ arylene, 4- to 6-membered monocyclic heterocycloalkylene, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, C ３ ~C ７ Monocyclic cycloalkylene, C ５ ~C １１ selected from bridged bicyclic cycloalkylene, -NH-NH-, and -NHC(O)NH-, wherein said 4-6 membered monocyclic heterocycloalkylene may optionally be substituted on a ring carbon atom with an oxo group; R ６ , R ７ and R ８ is H, -CH ２ CH ２ CH ２ NHC(O)NH ２ , benzyl, and a naturally occurring amino acid side chain; R ９ Ha-CH ２ NHC(O)-(CH ２ CH ２ O) ｑ -CH ３ and m is 0, 1 or 2; each occurrence of n is independently an integer from 0 to 4; q is selected from 4, 8, 10, and 12).

18. 17. The antibody-drug conjugate of claim 16, selected from the following: Table 3 or a pharmaceutically acceptable salt thereof, A is a 4- to 6-membered monocyclic heterocycloalkylene or a 5- to 11-membered bicyclic spirocyclic heterocycloalkylene; X is -(CH ２ ) ｎ -N(R ４ ) -, -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-C(O)R ５ -, -(CH ２ ) ｎ -N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ )-, 5- or 6-membered monocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -N(R ４ )-, 5- to 11-membered bicyclic spirocyclic heterocycloalkylene, -(CH ２ ) ｎ -R ５ -, (CH ２ ) ｎ -R ５ -N(R ４ )C(O)O-(CH ２ ) ｎ R ５ -N(R ４ ) -, -(CH ２ ) ｎ -NHC(O)O-(CH ２ ) ｎ -R ５ -N(R ４ ) -, -(CH ２ ) ｎ -R ５ -O-C(O)-N(R ４ )-(C １ ~C ６ alkylene)-N(R ４ ) -, -R ５ -NHC(O)CH ２ OCH ２ N (R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )C(O)NH-N(R ４ ) -, -R ５ -N(R ４ )C(O)CH ２ N (R ４ ) -, -(CH ２ ) ｎ -N(R ４ )-CH ２ -N(R ４ )- and -R ５ -O-CH ２ N (R ４ )—selected from; Y is a bond or —C(O)CH(R ７ ) NH—; Z is -C(O)CH(R ６ )NHC(O)CH(R ７ ) NH—; R ２ is H or C １ ~C ６ is alkyl; R ４ is H or C １～６ is alkyl; R ５ is C ６ ~C １０ arylene, 4- to 6-membered monocyclic heterocycloalkylene, C ３ ~C ７ Monocyclic cycloalkylene, C ５ ~C １１ selected from bridged bicyclic cycloalkylene, —NH—NH—, and —NHC(O)NH—; R ６ , R ７ and R ８ is H, -CH ２ CH ２ CH ２ NHC(O)NH ２ , benzyl, and a naturally occurring amino acid side chain; R ９ Ha-CH ２ NHC(O)-(CH ２ CH ２ O) ｑ -CH ３ and m is 0, 1 or 2; each occurrence of n is independently an integer from 0 to 4; q is selected from 4, 8, 10, and 12).

19. 19. The antibody-drug conjugate of claim 15, wherein L is an antibody.

20. (i) a compound of formula (XXII) 【Chemistry 15】 or a pharmaceutically acceptable salt thereof, R １’ is H or C １～６ is alkyl; R ２’ is a linker selected from linkers L-1 to L-12; Or, R １ and R ２ taken together with the common nitrogen atom to which they are each bonded, combine to form (i) a 3- to 7-membered monocyclic heterocycloalkyl group, (ii) a 5- to 11-membered bridged bicyclic heterocycloalkyl group, (iii) a 5- to 11-membered fused bicyclic heterocycloalkyl group, or (iv) a 5- to 11-membered spirocyclic heterocycloalkyl group, wherein said 3- to 7-membered monocyclic heterocycloalkyl group, said 5- to 11-membered bicyclic heterocycloalkyl group, said 5- to 11-membered fused heterocycloalkyl group, and said 5- to 11-membered spirocyclic heterocycloalkyl group are each optionally and independently selected from one or more R Ａ a linker selected from linkers L-1 through L-12, optionally substituted with a group, attached to ring atom (i), (ii), (iii), or (iv); R Ａ Each occurrence of C １～６ Alkyl, halo, -CN, -OR ３ , -N(R ３ ) ２ , C １ ~C ６ Aminoalkyl, -(C １ ~C ６ alkylene)-NHC(O)-(C １ ~C ６ aminoalkyl), —O—(C １～６ alkyl), —NHC(O)CH ２ OH, and -(C １ ~C ６ and (3- to 7-membered monocyclic heterocycloalkyl)-(3- to 7-membered alkylene)-(3- to 7-membered monocyclic heterocycloalkyl), wherein the 3- to 7-membered monocyclic heterocycloalkyl group is optionally and independently selected from one or more R Ｂ optionally substituted with a group; R Ｂ Each occurrence of C １～６ Alkyl, halo, -CN, -OR ３ , -N(R ３ ) ２ , C １ ~C ６ aminoalkyl, —O—(C １～６ alkyl), -NHC(O)-(C １ ~C ６ aminoalkyl), -(C １ ~C ６ alkylene)-NHC(O)-(C １ ~C ６ aminoalkyl), C １～６ Alkyl, halo, -CN, -OR ３ , -N(R ３ ) ２ , —O—(C １～６ alkyl), and —NHC(O)CH ２ OH); and (ii) a linker selected from: Table 4 【change】 and a linker-payload compound comprising: The linker is a linker that is a linker of the R ２ and the point of attachment on the linker is 【Chemistry 16】 A linker-payload compound represented by the formula:

21. A linker-payload compound having a formula selected from the following: Table 5 or a pharmaceutically acceptable salt thereof, wherein D is a payload selected from anti-cancer drugs.

22. 20. A pharmaceutical composition comprising: (a) the antibody-drug conjugate of any one of claims 15 to 19, or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier.

23. Use of (a) an antibody-drug conjugate of any one of claims 15 to 19, or a pharmaceutically acceptable salt thereof, or (b) a pharmaceutical composition of claim 22, for the manufacture of a medicament for treating or preventing cancer or tumors.

24. In those in need thereof, the following cancers are covered: breast cancer, ovarian cancer, cervical cancer, uterine cancer, prostate cancer, kidney cancer, urethral cancer, bladder cancer, liver cancer, stomach cancer, endometrial cancer, salivary gland cancer, esophageal cancer, melanoma, glioma, neuroblastoma, sarcoma, lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), colon cancer, rectal cancer, colorectal cancer, leukemia (e.g., acute lymphocytic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia), bone marrow cancer, 22. A method for treating or preventing a cancer selected from cancer, skin cancer, thyroid cancer, pancreatic cancer, and lymphoma (e.g., Hodgkin's lymphoma, non-Hodgkin's lymphoma, or recurrent anaplastic large cell lymphoma), comprising the step of administering a therapeutically effective amount of (a) the antibody-drug conjugate of any one of claims 15 to 19, or a pharmaceutically acceptable salt thereof, or (b) the pharmaceutical composition of claim 22, to a subject in need of such treatment.

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