Combination of MCL-1 inhibitor and anticancer agent

Abstract

METHODS OF TREATING CANCER BY ADMINISTERING MCL-1 INHIBITORS AND ANTI-CANCER AGENTS SOLUTION: The present disclosure relates generally to methods for treating cancer by administering an MCL-1 inhibitor and an anti-cancer agent. As is explained in detail in the present specification and as is specifically demonstrated, according to the present invention, a remarkable effect is achieved, which could not be easily expected by those skilled in the art from the prior art.SELECTED DRAWING: Figure 5

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Application No. 63 / 209,682, filed June 11, 2021, the contents of which are incorporated herein by reference in their entirety.

[0002] FIELD OF THE INVENTION This application relates generally to combination therapy of MCL-1 inhibitors with anti-cancer agents. [Background technology]

[0003] Apoptosis (programmed cell death) is a process for eliminating unwanted or potentially dangerous cells from an organism. Evasion of apoptosis is important for the development and sustained growth of tumors. Myeloid cell leukemia 1 protein (MCL-1; also abbreviated as Mcl-1 or MCL1) is an anti-apoptotic member of the Bcl-2 family of proteins. MCL-1 is overexpressed in many cancers. Overexpression of MCL-1 prevents cancer cells from undergoing apoptosis.

[0004] Studies have shown that MCL-1 inhibitors can be used to treat a variety of cancers. See, for example, "The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models," A. Kotschy et al., Nature, 2016(538):477-482; "Structure Based Design of "Non-Natural Peptidic Macrocyclic Mcl-1 Inhibitors", J.Johannes et al.,ACS Med.Chem.Lett.,2017,8(2):239-244&ACS Med.Chem.Lett.,2017,8(11):1204;"Synergistic action of the MCL-1 inhibitor S63845 with current therapies in preclinical models of triple-negative and HER2-amplified breast cancer", D.Merino et al.,Sci.Transl.Med.,2017 Aug.2,9(401):eaam7049;"Discovery of Mcl-1-specific inhibitor AZD5991 and preclinical activity in multiple myeloma "AMG 176,a Selective MCL1 Inhibitor,Is Effective in Hematologic Cancer Models Alone and in Combination with Established Therapies", S.Caenepeel et al.,Cancer Discov.,2018 Dec 8(12):1582-1597; "Discovery of S64315, a Potent and Selective Mcl-1 Inhibitor", Z. Szlavik at al., J. Med. Chem., 2020, 63(22): 13762-13795. There remains a need to provide more effective methods for the treatment of cancer. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] “The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models”, A. Kotschy et al., Nature, 2016(538):477-482 [Non-patent document 2] "Synergistic action of the MCL-1 inhibitor S63845 with current therapies in preclinical models of triple-negative and HER2-amplified breast cancer", D.Merino et al.,Sci.Transl.Med.,2017 Aug.2,9(401):eaam7049 [Non-patent document 3] "Discovery of Mcl-1-specific inhibitor AZD5991 and preclinical activity in multiple myeloma and acute myeloid leukemia", A. Tron et al., Nature Comm.2018(9):Article No.5341 [Non-patent document 4] "AMG 176,a Selective MCL1 Inhibitor,Is Effective in Hematologic Cancer Models Alone and in Combination with Established Therapies", S.Caenepeel et al.,Cancer Discov.,2018 Dec 8(12):1582-1597 [Non-Patent Document 5] "Discovery of S64315, a Potent and Selective Mcl-1 Inhibitor", Z. Szlavik at al., J. Med. Chem., 2020, 63(22): 13762-13795 Summary of the Invention [Means for solving the problem]

[0006] In some embodiments, provided herein are methods of treating cancer, comprising administering to a human patient in need thereof a therapeutically effective amount of an anti-cancer agent and a therapeutically effective amount of an MCL-1 inhibitor; The MCL-1 inhibitor is of formula (I) or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 1 is a 5-10 membered heteroaryl containing 1-2 heteroatoms; each heteroatom is independently selected from nitrogen, sulfur, and oxygen; R 1 The 5- to 10-membered heteroaryl is halo, hydroxyl, -CN, C 1~6 Alkyl, C 1~6 Haloalkyl, -OR a , and C 3~6 optionally substituted with 1 to 3 substituents independently selected from cycloalkyl; Each R 2 , R 3 , R 4 and R 5 are independently hydrogen or C 1~6 is alkyl; R 6 is hydrogen or halo; R a are independently hydrogen, C 1~6 Alkyl, C 2~6 Alkenyl, and C 3~10 It is cycloalkyl. [Brief explanation of the drawings]

[0007] [Figure 1] Paclitaxel treatment increases FBXW7 protein and decreases MCL1 protein and MCL1-BAK and MCL1-BIM protein dimers in TNBC cells.

[0008] [Figure 2] HCC70 inhibition and 95% CI synergy response surfaces.

[0009] [Figure 3] MDA-MB-468 inhibition and 95% CI synergy response surfaces.

[0010] [Figure 4] HCC1806 inhibition and 95% CI synergy response surfaces.

[0011] [Figure 5] TNBC PDX model CTG-1909 tumor growth.

[0012] [Figure 6] TNBC PDX model CTG-2010 tumor growth. DETAILED DESCRIPTION OF THE INVENTION

[0013] definition Unless the context requires otherwise, throughout this specification and claims, the word "comprise" and variations thereof, e.g., "comprises" or "comprising" are intended to be used in an open and inclusive sense, i.e., "including but not limited to." should be interpreted.

[0014] "C u~v ", i.e. (C u ~C v A prefix such as "C" indicates that the following group has u to v carbon atoms, where u and v are integers. 1~6 "Alkyl" indicates that the alkyl group has from 1 to 6 carbon atoms.

[0015] A dash ("-") that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -C(O)NH2 is attached through the carbon atom. Dashes at the front or end of a chemical group are for convenience, and chemical groups may be depicted with or without one or more dashes without losing their ordinary meaning. Unless chemically or structurally required, no directionality is indicated or implied by the order in which chemical groups are written or named.

[0016] The term "substituted" means that one or more hydrogen atoms on a hydrocarbon have been replaced with one or more atoms or groups other than hydrogen, provided that the normal valence of the designated carbon atom is not exceeded. A "substituent" is an atom or group that replaces a hydrogen atom on a hydrocarbon when "substituted." Unless otherwise specified, if a group is described as optionally substituted, any substituents of the group are themselves unsubstituted.

[0017] Reference herein to "about" a value or parameter includes (and describes) embodiments that relate to the value or parameter itself. In certain embodiments, the term "about" includes the stated amount ±10%. In other embodiments, the term "about" includes the stated amount ±5%. In certain other embodiments, the term "about" includes the stated amount ±1%. Also, for that term, "about X" includes the description of "X." Additionally, the singular forms "a" and "the" include plural references unless the context clearly indicates otherwise. Thus, for example, reference to "a compound" includes a plurality of such compounds, and reference to "an assay" includes reference to one or more assays and equivalents thereof known to those of skill in the art.

[0018] "Alkyl" refers to an unbranched or branched saturated hydrocarbon chain. In the case of alkyl, alkyl has 1 to 20 carbon atoms (i.e., C 1~20 alkyl), having 1 to 12 carbon atoms (i.e., C 1~12alkyl), having 1 to 8 carbon atoms (i.e., C 1~8 alkyl), having 1 to 6 carbon atoms (i.e., C 1~6 alkyl), having 1 to 4 carbon atoms (i.e., C 1~4 alkyl), having 1 to 3 carbon atoms (i.e., C 1~3 alkyl), or having 1 to 2 carbon atoms (i.e., C 1~2 alkyl). Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl group having a specific number of carbons is designated by a chemical name or identified by a molecular formula, all positional isomers having that number of carbons can be included; thus, for example, "butyl" includes n-butyl (i.e., -(CH2)3CH3), sec-butyl (i.e., -CH(CH3)CH2CH3), isobutyl (i.e., -CH2CH(CH3)2), and tert-butyl (i.e., -C(CH3)3), and "propyl" includes n-propyl (i.e., -(CH2)2CH3) and isopropyl (i.e., -CH(CH3)2).

[0019] "Aryl" refers to an aromatic carbocyclic group having a single ring (e.g., monocyclic) or multiple rings (e.g., bicyclic or tricyclic), including fused systems. As used herein, aryl refers to a group having 6 to 20 ring carbon atoms (i.e., C6-20 aryl), 6 to 12 carbon ring atoms (i.e., C6-12 aryl), or 6 to 10 carbon ring atoms (i.e., C6-10 aryl). Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, anthryl, and the like. However, aryl does not in any way encompass or overlap with heteroaryl, as defined below. When one or more aryl groups are fused to a heteroaryl ring, the resulting ring system is heteroaryl.

[0020] "Cycloalkyl" refers to saturated or partially saturated cyclic alkyl groups having single or multiple rings, including fused, bridged, and spirocyclic systems. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0021] As used herein, "halo" or "halogen" refers to fluoro (-F), chloro (-Cl), bromo (-Br), and iodo (-I).

[0022] As used herein, the term "haloalkyl" refers to an alkyl, as defined herein, in which one or more hydrogen atoms of the alkyl are independently replaced with halogen substituents, which may be the same or different. For example, C 1~6 Haloalkyl is C 1~6 alkyl, C 1~6 One or more of the hydrogen atoms of the alkyl is replaced with a halo substituent. Examples of haloalkyl groups include, but are not limited to, fluoromethyl, fluorochloromethyl, difluoromethyl, difluorochloromethyl, trifluoromethyl, 1,1,1-trifluoroethyl, and pentafluoroethyl.

[0023] "Heteroaryl" refers to aromatic groups, including groups having aromatic tautomers or resonance structures, having monocyclic, polycyclic, or multiple fused rings with at least one heteroatom in the ring, i.e., one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur, where the nitrogen or sulfur may be optionally oxidized. Thus, the term includes rings having one or more cyclic O, N, S, S(O), S(O)2, and N-oxide groups. The term includes rings having one or more cyclic C(O) groups. As used herein, heteroaryl refers to heteroaryls having 5 to 20 ring atoms (i.e., 5-20 membered heteroaryls), 5 to 12 ring atoms (i.e., 5-12 membered heteroaryls), or 5 to 10 ring atoms (i.e., 10-12 membered heteroaryls). Heteroaryl groups include aryl, ...

[0024] The terms "heterocyclyl," "heterocycle," or "heterocyclic" refer to a monoradical or diradical saturated or unsaturated group having a single ring or multiple fused rings with one or more heteroatoms selected from nitrogen, sulfur, phosphorus, and / or oxygen within the ring. The heteroatoms within a "heterocyclyl" can be oxidized, e.g., -N(O)-, -S(O)-, -S(O)2-. A heterocyclyl can be monocyclic or polycyclic, and the polycyclic rings can be fused, bridged, or spiro.

[0025] "Isomers" are different compounds that have the same molecular formula. Isomers include stereoisomers, enantiomers, and diastereomers.

[0026] "Stereoisomers" refer to compounds made up of the same atoms connected by the same bonds but with different, not interchangeable, three-dimensional structures. The present disclosure contemplates various stereoisomers and mixtures thereof, and includes "enantiomers," which refer to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0027] "Tautomer" refers to a proton migration from one atom of a molecule to another atom of the same molecule. The present disclosure includes tautomers of any such compounds.

[0028] A "solvate" is formed by the interaction of a solvent and a compound. Solvates of the salts of the compounds described herein are also provided. Hydrates of the compounds described herein are also provided.

[0029] As used herein, the term "prodrug" refers to a biologically inactive derivative of a drug that, upon administration to the human body, is converted into the biologically active parent drug by some chemical or enzymatic pathway.

[0030] "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The symbol "(±)" is used to designate a racemic mixture where appropriate.

[0031] "Diastereomers" are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.

[0032] As used herein, "treatment" or "treating" is an approach for obtaining a beneficial or desired result. For purposes of this disclosure, beneficial or desired results include, but are not limited to, alleviating symptoms and / or reducing the symptoms associated with a disease or condition. In one embodiment, "treatment" or "treating" includes one or more of: a) inhibiting a disease or condition (e.g., reducing one or more symptoms resulting from a disease or condition and / or reducing the severity of the disease or condition); b) delaying or halting the onset of one or more symptoms associated with a disease or condition (e.g., stabilizing the disease or condition, slowing the worsening or progression of the disease or condition); and c) alleviating the disease or condition, e.g., regressing clinical symptoms, improving the disease state, slowing disease progression, increasing quality of life, and / or prolonging survival.

[0033] As used herein, "prevention" or "preventing" refers to a regimen that protects against the onset of a disease or disorder, such that clinical symptoms of the disease or disorder do not develop. Thus, "prevention" relates to the administration of a treatment to a subject before symptoms of the disease are detectable in the subject. The subject may be an individual at risk of developing a disease or disorder, such as an individual with one or more risk factors known to be associated with the onset or development of a disease or disorder.

[0034] As used herein, the term "therapeutically effective amount" or "effective amount" refers to an amount effective to elicit a desired biological or medical response when administered to a subject to treat a disease, including an amount of an agent sufficient to effect such treatment of the disease. The effective amount will vary depending on the particular agent and characteristics of the treated subject, such as age, weight, etc. An effective amount can include a range of amounts. As understood in the art, an effective amount can be one or more doses, i.e., a single dose or multiple doses may be required to achieve a desired therapeutic endpoint. An effective amount may be considered in relation to the administration of one or more therapeutic agents, and a single agent may be considered to be administered in an effective amount when a desired or beneficial result can be or is achieved in combination with one or more other agents. The appropriate dose of any co-administered agent may optionally be reduced due to the combined action (e.g., additive or synergistic) of the agents.

[0035] As used herein, "co-administration" includes administering a unit dose of an agent disclosed herein before or after administering a unit dose of one or more additional therapeutic agents, e.g., administering an agent disclosed herein within seconds, minutes, or hours of administering one or more additional therapeutic agents. For example, in some embodiments, a unit dose of an agent disclosed herein is administered first, followed within seconds or minutes by a unit dose of one or more additional therapeutic agents. Alternatively, in other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed within seconds or minutes by a unit dose of a compound disclosed herein. In some embodiments, a unit dose of a compound disclosed herein is administered first, followed several hours (e.g., 1-12 hours) later by a unit dose of one or more additional therapeutic agents. In other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed several hours (e.g., 1-12 hours) later by a unit dose of a compound disclosed herein.

[0036] Administration "in combination with" one or more further therapeutic agents includes simultaneous (concurrent) and sequential or sequential administration in any order.

[0037] The term "concurrently" is used herein to refer to the administration of two or more therapeutic agents, where at least a portion of the administration overlaps in time, or the administration of one therapeutic agent is within a short period of time relative to the administration of another therapeutic agent, e.g., the two or more therapeutic agents are administered within a certain time interval of no more than a certain number of minutes.

[0038] The term "sequentially" is used herein to refer to the administration of two or more therapeutic agents, where the administration of one or more agents continues after the administration of one or more other agents is discontinued, or where the administration of one or more agents begins before the administration of one or more other agents, e.g., where the two or more therapeutic agents are administered at intervals of more than a certain number of minutes.

[0039] As used herein, "in conjunction with" refers to the administration of one therapeutic modality in addition to another therapeutic modality. Thus, "in conjunction with" refers to the administration of one therapeutic modality before, during, or after the administration of another therapeutic modality to an individual.

[0040] The term "conjugate" or "antibody-drug conjugate" refers to an antibody that is chemically linked to a second chemical moiety, such as a therapeutic or cytotoxic agent. includes a chemical compound, a mixture of chemical compounds, a biological macromolecule, or an extract made from biological materials. In some embodiments, therapeutic or cytotoxic agents include, but are not limited to, pertussis toxin, taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, and analogs or homologs thereof. When used in the context of an immunoassay, the conjugated antibody can be a detectably labeled antibody used as a detection antibody.

[0041] "Intravenous administration" is the administration of a substance into a vein or "into a vein." Compared to other routes of administration, the intravenous (IV) route is a more common method for delivering fluids and medications throughout the body. It is a faster method. Infusion pumps can allow precise control over the flow rate and total amount of medication delivered. However, if changes in flow rate do not have serious consequences or a pump is not available, the infusion is often left to flow by simply placing the bag above the patient's level and adjusting the rate using a clamp. Alternatively, if the patient requires a high flow rate and the IV access device is large enough in diameter to accommodate it, a rapid infuser can be used. This is either an inflatable cuff placed around the fluid bag to force the fluid into the patient, or a similar electrical device that can also heat the infused fluid. If the patient only needs medication at a specific time, an intermittent infusion is used, which does not require additional fluid. This can use the same technique as an intravenous infusion (pump or gravity infusion), but after the full dose of medication is given, the tubing is disconnected from the IV access device. Some medications are also given by IV push or bolus; that is, a syringe is connected to the IV access device and the medication is injected directly (or slowly, if this stimulates the vein or causes a rapid effect). When a medication is injected into the fluid stream of an IV tubing, there must be some means to ensure that it reaches the patient through the tubing. Typically, this is accomplished by allowing the fluid stream to flow normally, thereby carrying the medication into the bloodstream. However, a second fluid injection is sometimes used after the injection as a "washout" to allow the medication to flow more quickly into the bloodstream. Thus, in one embodiment, the agent(s) or combination of agents described herein may be administered by IV administration, either alone or in combination with administration of specific components of the treatment regimen by oral or parenteral routes.

[0042] "Oral administration" means a route of administration in which a substance is taken through the mouth, and includes buccal, sublabial, and sublingual administration, as well as enteral administration and administration through the respiratory tract, unless the drug is in direct contact with any of the oral mucosa, e.g., via a tube. Typical forms for oral administration of therapeutic agents include the use of tablets or capsules. Thus, in one embodiment, a compound(s) or combination of compounds described herein may be administered by the oral route, either alone or in combination with administration of certain components of the treatment regimen by IV or parenteral routes.

[0043] Also provided herein are pharmaceutically acceptable salts, hydrates, solvates, tautomers, polymorphs, and prodrugs of the compounds of Formula (I) described herein. "Pharmaceutically acceptable" or "physiologically acceptable" refers to compounds, salts, compositions, dosage forms, and other materials that are suitable for use in human medicine.

[0044] The compounds of formula (I) described herein may be prepared and / or formulated as pharmaceutically acceptable salts. Pharmaceutically acceptable salts are non-toxic salts of the free base form of the compounds that possess the desired pharmacological activity of the free base. These salts may be prepared from inorganic or organic acids or from inorganic or organic acids. Pharmaceutically acceptable salts may be derived from inorganic bases. For example, compounds containing a basic nitrogen can be prepared as pharmaceutically acceptable salts by contacting the compound with an inorganic or organic acid. Non-limiting examples of pharmaceutically acceptable salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyl phosphate ... Examples of suitable pharmaceutically acceptable salts include benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, methylsulfonate, propylsulfonate, besylate, xylenesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate, and mandelate. A list of other suitable pharmaceutically acceptable salts can be found in Remington: The Science and Practice of Pharmacy, 21 st Edition, Lippincott Williams and Wilkins, Philadelphia, Pa., 2006.

[0045] Non-limiting examples of "pharmaceutically acceptable salts" of the compounds of formula (I) disclosed herein also include alkali metal (e.g., sodium, potassium), alkaline earth metal (e.g., magnesium), ammonium, and NX4 salts. + wherein X is C1-C4 alkyl. Base addition salts such as sodium or potassium salts are also included. MCL-1 inhibitors compound

[0046] In some embodiments, provided herein are methods of treating cancer, comprising administering to a human patient in need thereof a therapeutically effective amount of an anti-cancer agent and a therapeutically effective amount of an MCL-1 inhibitor; The MCL-1 inhibitor is of formula (I) or a pharmaceutically acceptable salt thereof: [ka] In the formula, R 1 is a 5-10 membered heteroaryl containing 1-2 heteroatoms; each heteroatom is independently selected from nitrogen, sulfur, and oxygen; R 1 The 5- to 10-membered heteroaryl is halo, hydroxyl, -CN, C 1~6 Alkyl, C 1~6 Haloalkyl, -OR 7 , and C 3~6 optionally substituted with 1 to 3 substituents independently selected from cycloalkyl; Each R 2 , R 3 , R 4 and R 5 are independently hydrogen or C 1~6 is alkyl; R 6 is hydrogen or halo; R 7 are independently hydrogen, C 1~6 Alkyl, C 2~6 Alkenyl, and C 3~10 It is cycloalkyl.

[0047] In some embodiments of the methods described herein, the MCL-1 inhibitor is a compound of formula (II), or a pharmaceutically acceptable salt thereof: [ka] Each R 1 , R 2 , R 3 , R 4 , R 5 , and R 6is provided herein as a method as defined above or elsewhere in this disclosure.

[0048] In some embodiments, the MCL-1 inhibitor is a compound of formula (III): [ka] or a pharmaceutically acceptable salt thereof:

[0049] In some embodiments, the MCL-1 inhibitor is a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, wherein R 2 is hydrogen. In some embodiments, R 2 is C 1~3 R is alkyl. 2 is methyl.

[0050] In some embodiments, the MCL-1 inhibitor is a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, wherein R 3 is C 1~3 In some embodiments, R 3 is methyl.

[0051] In some embodiments, the MCL-1 inhibitor is a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, wherein R 4 is hydrogen. In some embodiments, R 5 is C 1~3 In some embodiments, R 5 is methyl. In some embodiments, R 6 is Cl.

[0052] In some embodiments, the MCL-1 inhibitor is a compound of Formula (I), Formula (II), or Formula (III), or a pharmaceutically acceptable salt thereof, wherein R 1 is C 1~4 Alkyl and C 1~4 Optionally substituted with alkoxyl [ka] In some embodiments, R 1 is optionally substituted with -CH3 and -OCH3 [ka] In some embodiments, R 1 is substituted with -CH3 and -OCH3 [ka] In some embodiments, R 1 teeth [ka] is.

[0053] In some embodiments, the MCL-1 inhibitor is compound A, N-[(4S,7aR,9aR,10S,11E,14S)-6-chloro-10-methoxy-14-methyl-16-oxide-18-oxo-3′,4′,7,7a,8,9,9a,10,13,14,15,18-dodecahydro-2′H-spiro[1,19-(ethanediylidene)-16λ] 4 -cyclobuta[i][1,4]oxazepino[3,4-f][1,2,7]thiadiazacyclohexadecin-4,1′-naphthalen]-16-yl]-3-methoxy-1-methyl-1H-pyrazole-4-carboxamide, having the following structure: [ka] Compound A is described in USPN 10,703,733 and Example 154 of WO2019 / 222112, which are incorporated herein by reference.

[0054] In some embodiments, MCL-1 inhibitors that can be administered include those disclosed in U.S. Pat. No. 10,703,733 (Gilead Sciences), AMG-397, AMG-176, PRT-1419, S64315, AZD59991, ABBV-467, WO 2019222112 (Gilead Sciences), WO 2021096860 (Gilead Sciences), and others. 2017147410 (Amgen), 2019046150 (Amgen), 2019036575 (Amgen), 2021021259 (Amgen), 2019173181 (Amgen), 2018183418 (Amgen), 2016033486 (Amgen), 2018178226 (AstraZeneca), 201718 No. 2625 (AstraZeneca), No. 2018178227 (AstraZeneca), No. 2020099470 (AstraZeneca), No. 2019211721 (AstraZene) ca), No. 2020097577 (Prelude), No. 2020123994 (Prelude), No. 2008104386 (AbbVie), No. 2008104385 (AbbVie), No. 2 No. 008131000 (AbbVie), No. 2008130970 (AbbVie), No. 2019035911 (AbbVie), No. 2019 No. 035927 (AbbVie), No. 2019035899 (AbbVie), No. 2010049816 (Servier), No. 202016 No. 0157 (Servier), No. 2020115183 (Servier), No. 2020099542 (Servier), No. 2015097123 (Servier), No. 2018078064 (Servier), No. 2020254299 (Servier), No. 201812 7575 (Servier), 2018234433 (Servier), 018015526 (Servier), 2016207225 (Servier), 2020078875 (Servier), 2017125224 (Servier), 2020236817 (Servier), 2016207226 (Servier), 2016207217 (Servier), 2016207216 (Servier), and 2007147613 (Novartis).

[0055] In some embodiments, the MCL-1 inhibitor is selected from AMG-397, AMG-176, PRT-1419, and S64315. In some embodiments, the MCL-1 inhibitor is AMG-176. In some embodiments, the MCL-1 inhibitor is AMG-397. In some embodiments, the MCL-1 inhibitor is PRT-1419. In some embodiments, the MCL-1 inhibitor is S64315.

[0056] The compounds disclosed herein may contain one or more asymmetric centers and may therefore give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)-. The present disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-) or (R)- and (S)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high-pressure liquid chromatography (HPLC). Likewise, all tautomeric forms are also intended to be included. formulation

[0057] In the methods provided herein, the MCL-1 inhibitor can be administered as a pharmaceutical composition. In certain embodiments, the pharmaceutical composition comprises a compound of Formula (I), (II), (III) or Compound A, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises one or more additional therapeutic agents, as described more fully below.

[0058] Pharmaceutical compositions comprising the MCL-1 inhibitors disclosed herein, or pharmaceutically acceptable salts thereof, may be prepared using one or more pharmaceutically acceptable excipients, which may be selected according to ordinary practice. A "pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, filler, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier approved by the U.S. Food and Drug Administration as acceptable for use in humans or veterinary animals.

[0059] In certain embodiments, the pharmaceutical composition is provided as a solid dosage form, including a solid oral dosage form such as a tablet. Tablets may contain excipients including lubricants, fillers, binders, etc. Aqueous Compositions The compositions may be prepared in sterile form and, if intended for delivery by other than oral administration, may generally be isotonic. All compositions are prepared in accordance with the methods set forth in Rowe et al., Handbook of Pharmaceutical Excipients, 6 th The composition may also contain excipients such as those described in the American Pharmacists Association, 2009 edition. Excipients may include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextrin, hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid, and the like.

[0060] The pharmaceutical compositions disclosed herein include those suitable for various routes of administration, including oral administration. The compositions may be presented in unit dosage form and may be prepared by any of the methods known in the art of pharmacy. Such methods include the step of bringing into association the active ingredient (e.g., a compound of the present disclosure or a pharmaceutical salt thereof) with one or more pharmaceutically acceptable excipients. The compositions may be prepared by uniformly and intimately bringing into association the active ingredient with liquid excipients or finely divided solid excipients, or both, and then, if necessary, shaping the product. Techniques and formulations are generally described in Remington: The Science and Practice of Pharmacy, 21 st Edition, Lippincott Williams and Wilkins, Philadelphia, Pa., 2006.

[0061] Compositions described herein suitable for oral administration may be presented as discrete units (unit dosage forms) including, but not limited to, capsules, cachets, or tablets, each containing a predetermined amount of the active ingredient. In one embodiment, the pharmaceutical composition is a tablet.

[0062] In some embodiments, the tablets contain Compound A in strengths of 5 mg and 25 mg. In some embodiments, the tablets contain copovidone, lactose monohydrate, microcrystalline cellulose, crospovidone, magnesium stearate, polyvinyl alcohol, titanium dioxide, polyethylene glycol, and talc. Antibody-drug conjugates (ADCs)

[0063] In some embodiments, the methods of treating cancer disclosed herein comprise administering to a human patient in need thereof a therapeutically effective amount of an anti-cancer agent, a therapeutically effective amount of an MCL-1 inhibitor, and a therapeutically effective amount of an antibody-drug conjugate. In some embodiments, the antibody-drug conjugate comprises an anti-Trop-2 antibody and an anti-cancer agent.

[0064] In some embodiments, the antibody-drug conjugate is sacituzumab govitecan, which is disclosed in U.S. Patent No. 7,999,083. In some embodiments, the ADC comprises an antibody-drug conjugate disclosed in U.S. Patent No. 7,999,083, which is incorporated herein by reference. In some embodiments, the sacituzumab govitecan is sacituzumab govitecan-hziy.

[0065] In some embodiments, the anti-Trop-2 antibody-drug conjugate is datopotamab deruxtecan. In some embodiments, anti-Trop-2 antibody-drug conjugates that can be administered include, but are not limited to, those disclosed in U.S. Patent Nos. 9,850,312, 9,850,312, WO 20240467, and WO 18036438.

[0066] In some embodiments, the antibody portion of the ADC is an IgG antibody or antigen-binding antibody fragment. The antibody can be of various isotypes, preferably human IgG1, IgG2, IgG3, or IgG4, more preferably containing human IgG1 hinge and constant region sequences. The antibody or fragment thereof can be human-mouse chimeric, human-primate chimeric, human The antibodies can be monoclonal (human framework and mouse hypervariable (CDR) regions), or fully human anti-IgG4 antibodies, as well as variants thereof, such as half-IgG4 antibodies (called "unibodies") (van der Neut Kolfschoten et al. (Science 2007;317:1554-1557). More preferably, the antibody or fragment thereof may be designed or selected to contain a human constant region 65 sequence belonging to a particular allotype, which may result in reduced immunogenicity when the antibody or ADC is administered to a human subject. Preferred allotypes for administration include non-Glml allotypes (nGlml), such as Glm3, Glm3,1, Glm3,2, or Glm3,1,2. More preferably, the allotype is selected from the group consisting of nGlml, Glm3, nGlml,2, and Km3 allotypes.

[0067] In some embodiments, the antibody moiety of the ADC is an anti-Trop-2 antibody, such as TROP2-XPAT (Amunix), BAT-8003 (Bio-Thera Solutions), TROP-2-IR700 (Chiome Bioscience), datopotamab deruxtecan (Daiichi Sankyo, AstraZeneca), GQ-1003 (Genequantum Healthcare, Samsung BioLogics), DAC-002 (Hangzhou DAC Biotech, Shanghai Junshi Biosciences), sacituzumab govitecan (Gilead Sciences), E1-3s (Immunomedics / Gilead, IBC Pharmaceuticals), TROP2-TRACTr (Janux Therapeutics), or LIV-2008 (LivTech / Chiome, Yakult Honsha, Shanghai Henlius) BioTech), LIV-2008b (LivTech / Chiome), anti-TROP-2a (Oncoxx), anti-TROP-2b (Oncoxx), OXG-64 (Oncoxx), OXS-55 (Oncoxx), humanized anti-Trop2-SN38 antibody conjugate (Shanghai Escugen Biotechnology, TOT Biopharma), anti-Trop2 antibody-CLB-SN-38 conjugate (Shanghai Fudan-Zhangjiang Bio-Pharmaceutical), SKB-264 (Sichuan Kelun Pharmaceutical / Klus Pharma), TROP2-Ab8 (Abmart), Trop2-IgG (Nanjing Medical University (NMU)), 90Y-DTPA-AF650 (Peking University First Hospital), hRS7-CM (SynAffix), 89Zr-DFO-AF650 (University of Wisconsin-Madison), anti-Trop2 antibody (Mediterranea Theranostic, LegoChem Biosciences), and KD-065 (Nanjing KAEDI Biotech).

[0068] Further examples of anti-TROP-2 therapeutics include, but are not limited to, E1.BB.3z-92MI (Immunomedics / Gilead), anti-Trop-2 CAR-T (Gilead), Trop-2CAR-T (Hangzhou Lonzyme Biological Technology), ARB-001 (Arbele), and MT-103 (Myeloid Therapeutics).

[0069] Examples of anti-TROP-2 antibodies include those disclosed in International Publication Nos. 2020016662 (Abmart), 2020249063 (Bio-Thera Solutions), U.S. Patent Application Publication No. 20190048095 (Bio-Thera Solutions), WO 2013077458 (LivTech / Chiome), EP 20110783675 (Chiome), WO 2015098099 (Daiichi Sankyo), and WO 2017002776 (Daiichi Sankyo). ), US Patent Application Publication No. 2020130125 (Daiichi Sankyo), US Patent Application Publication No. 2020240467 (Daiichi Sankyo), US Patent Application Publication No. 2021093730 (Daiichi Sankyo), US Patent No. 9850312 (Daiichi Sankyo), Chinese Patent No. 112321715 (Biosion), U.S. Patent Application Publication No. 2006193865 (Immunomedics / Gilead), International Publication No. 2011068845 (Immunomedics / Gilead), U.S. Patent Application Publication No. 2016296633 (Immunomedics / Gilead), International Publication No. 2017021017 (Immunomedics / Gilead), International Publication No. 2017209594 No. (Immunomedics / Gilead), No. 2017274093 (Immunomedics / Gilead), No. 2018110772 (Immunomedics / Gilead), No. 2018185351 (Immunomedics / Gilead), No. 2018271992 (Immunomedics / Gilead), International Publication No. 2018217227 (Immunomedics / Gilead), US Chinese Patent Application Publication No. 2019248917 (Immunomedics / Gilead), Chinese Patent No. 111534585 (Immunomedics / Gilead), US Patent Application Publication No. 2021093730 (Immunomedics / Gilead), US Patent No. 2021069343 (Immunomedics / Gilead), US Patent No. 8435539 (Immunomedics / Gilead), US Patent No. 8435529 (Immunomedics / Gilead), Immunomedics / Gilead), US Patent No. 9492566 (Immunomedics / Gilead), International Publication No. 2003074566 (Gilead), International Publication No. 2020257648 (Gilead), US Patent Application Publication No. 2013039861 (Gilead), International Publication No. 2014163684 (Gilead), US Patent No. 9427464 (LivTech / Chiome), US Patent No. 10501555 (Abruzzo Theranostic / Oncoxx), WO 2018036428 (Sichuan Kelun Pharma), WO 2013068946 (Pfizer), WO 2007095749 (Roche), and WO 2020094670 (SynAffix).

[0070] Further examples of anti-TROP-2 therapeutic agents include, but are not limited to, those described in WO 2016201300 (Gilead) and CN 108440674 (Hangzhou Lonzyme Biological Technology).

[0071] In some embodiments, the anti-Trop-2 antibody is selected from hRS7, Trop-2-XPAT, and BAT-8003.

[0072] In some embodiments, the anti-Trop-2 antibody is hRS7, as disclosed in U.S. Patent Nos. 7,238,785, 7,517,964, and 8,084,583, which are incorporated herein by reference.

[0073] In some embodiments, the antibody-drug conjugate comprises an anti-Trop-2 antibody and an anti-cancer drug joined by a linker. In some embodiments, the linker includes a linker disclosed in U.S. Patent No. 7,999,083. In some embodiments, the linker is CL2A.

[0074] In some embodiments, the drug moiety of the antibody-drug conjugate is a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is doxorubicin (DOX), Epirubicin, morpholinodoxorubicin (morpholino-DOX), cyanomorpholino- Selected from doxorubicin (cyanomorpholino-DOX), 2-pyrrolino-doxorubicin (2-PDOX), CPT, 10-hydroxycamptothecin, SN-38, topotecan, lutotecan, 9-aminocamptothecin, 9-nitrocamptothecin, taxanes, geldanmycin, ansamycins, and epothilones. In some embodiments, the chemotherapeutic moiety is SN-38. formulation

[0075] Suitable routes of administration of ADCs include, but are not limited to, oral, parenteral, subcutaneous, rectal, transmucosal, intestinal administration, intramuscular, intramedullary, intrathecal, direct intracerebroventricular, intravenous, intravitreal, intraperitoneal, intranasal, or intraocular injection. Alternatively, the compound may be administered locally rather than systemically, for example, by injecting the compound directly into a solid tumor.

[0076] The ADC can be formulated according to known methods for preparing pharmaceutically useful compositions, whereby the ADC is combined in a mixture with a pharmaceutically suitable excipient. The ADC can be formulated for intravenous administration, for example, by bolus injection, slow infusion, or continuous infusion. In some embodiments, the antibody is infused over a period of less than about 4 hours. In some embodiments, the antibody is infused over a period of less than about 3 hours. For example, the first 25-50 mg can be infused within 30 minutes or 15 minutes, with the remainder infused over the next 2-3 hours. Injectable formulations can be provided in unit dosage form, for example, in ampoules or in multi-dose containers, with preservatives added. The compositions can take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending agents, stabilizers, and / or dispersing agents. Alternatively, the active ingredient can be in powder form for constitution with a suitable vehicle, for example, pyrogen-free distilled water, before use. Treatment method

[0077] In some embodiments, the present disclosure provides a combination of MCL-1 and an anti-cancer agent for treating cancer. In some embodiments, the MCL-1 inhibitor is Compound A.

[0078] In some embodiments, the cancer is a Trop-2-expressing cancer.

[0079] In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a solid malignant tumor. In some embodiments, the cancer is an advanced solid malignant tumor.

[0080] In some embodiments, the cancer is selected from breast cancer, cervical cancer, colorectal cancer, endometrial cancer, epithelial ovarian cancer, esophageal cancer, follicular thyroid cancer, gastric cancer or gastroesophageal junction adenocarcinoma, head and neck cancer, lung cancer, hepatocellular carcinoma, non-small cell lung cancer, ovarian cancer, prostate cancer, renal cell carcinoma, small cell lung cancer, urothelial cancer, and urinary tract cancer.

[0081] In some embodiments, the cancer is selected from triple-negative breast cancer (TNBC), HR+ / HER2- breast cancer, urothelial cancer, non-squamous non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), head and neck squamous cell carcinoma (HNSCC), and muscle-invasive bladder cancer (MIBC).

[0082] In some embodiments, the cancer is metastatic. In some embodiments, the cancer is refractory.

[0083] In some embodiments, the cancer is selected from the group consisting of metastatic non-squamous non-small cell lung cancer (mNSCLC), metastatic triple-negative breast cancer (mTNBC), and metastatic non-small cell lung cancer (mTNBC) with non-specific histology. Selected from soft tissue sarcomas.

[0084] In some embodiments, the cancer is metastatic non-squamous non-small cell lung cancer (mNSCLC). In some embodiments, the cancer is metastatic triple-negative breast cancer (mTNBC). In some embodiments, the cancer is metastatic soft tissue sarcoma with non-specific histology.

[0085] In some embodiments, the human patient has received at least one other therapy prior to treatment with the combination therapy of an MCL-1 inhibitor and an anticancer conjugate. In some embodiments, the human patient has failed another therapy prior to treatment disclosed herein. In some embodiments, the human patient has failed one chemotherapy.

[0086] In some embodiments, the human patient has failed therapy with an anti-PD1 agent or an anti-PDL1 agent prior to treatment with the combination therapy of an MCL-1 inhibitor and an anti-cancer agent.

[0087] In some embodiments, the MCL-1 inhibitor and the anti-cancer agent are administered simultaneously or separately.

[0088] In some embodiments, the MCL-1 inhibitor is Compound A. In general, the dosage of Compound A administered to a human will vary depending on factors such as the patient's age, weight, height, sex, general medical condition, and medical history. It may be desirable to provide the recipient with a dose of antibody-conjugate ranging from about 1 mg / kg to 24 mg / kg as a single intravenous infusion, although lower or higher doses may be administered depending on the circumstances. For example, a dose of 1 to 20 mg / kg for a 70 kg patient would be 70 to 1,400 mg. Dosages may be repeated as needed, for example, once weekly for 4 to 10 weeks, once weekly for 8 weeks, or once weekly for 4 weeks. Maintenance therapy may also be administered less frequently, for example, every other week for several months, or monthly or quarterly for many months. In some embodiments, the dosages include 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 22 mg / kg, 24 mg / kg, 26 mg / kg, 28 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 55 mg / kg Doses include, but are not limited to, 65 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 120 mg / kg, 140 mg / kg, 150 mg / kg, 160 mg / kg, 180 mg / kg, 200 mg / kg, 220 mg / kg, 240 mg / kg, 250 mg / kg, 260 mg / kg, 280 mg / kg, 300 mg / kg, 350 mg / kg, 400 mg / kg, 450 mg / kg, 550 mg / kg, 600 mg / kg, 650 mg / kg, 700 mg / kg, 750 mg / kg, and 800 mg / kg. Any amount in the range of 1 to 300 mg / kg may be used. Any amount in the range of 1 to 100 mg / kg may be used. In some embodiments, the dose is administered multiple times, once or twice weekly.In some embodiments, dosage may utilize a minimum dosing schedule of 4 weeks, 8 weeks, 16 weeks, or longer. The dosing schedule may include once or twice weekly administration in a cycle selected from the group consisting of: (i) weekly, (ii) every other week, (iii) 1 week on treatment followed by 2, 3, or 4 weeks rest, (iv) 2 weeks on treatment followed by 1, 2, 3, or 4 weeks rest, (v) 3 weeks on treatment followed by 1, 2, 3, 4, or 5 weeks rest, (vi) 4 weeks on treatment followed by 1, 2, 3, 4, or 5 weeks rest, (vii) 5 weeks on treatment followed by 1, 2, 3, 4, or 5 weeks rest; and (viii) monthly. The cycle may be repeated 4, 6, 8, 10, 12, 16, 20, or more times. good.

[0089] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally.

[0090] In some embodiments, the amount of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered at a dosage of about 5 mg / kg, 15 mg / kg, or 50 mg / kg.

[0091] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered at a dosage of about 5 mg / kg.

[0092] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in a 21 day cycle with 2 days on medication followed by 5 days off medication.

[0093] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered on days 1, 2, 8, 9, 15, and 16 of each 21-day cycle for up to 105 weeks.

[0094] In some embodiments, the anti-cancer drug is administered as an intravenous infusion.

[0095] In general, the dosage of an anticancer drug administered to a human will vary depending on factors such as the patient's age, weight, height, sex, general medical condition, and medical history. It may be desirable to provide the recipient with a dose of antibody-conjugate ranging from about 1 mg / kg to 24 mg / kg as a single intravenous infusion, although lower or higher dosages may be administered depending on the circumstances. For example, a dosage of 1 to 20 mg / kg for a 70 kg patient would be 70 to 1,400 mg. Dosages can be repeated as needed, for example, once weekly for 4 to 10 weeks, once weekly for 8 weeks, or once weekly for 4 weeks. Maintenance therapy may also be administered less frequently, for example, every other week for several months, or monthly or quarterly for many months. In some embodiments, dosages include, but are not limited to, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 22 mg / kg, and 24 mg / kg. Any amount in the range of 1 to 24 mg / kg may be used. In some embodiments, dosages are administered multiple times, once or twice weekly. Minimum dosing schedules of 4 weeks, 8 weeks, 16 weeks, or longer may be used. The dosing schedule may comprise administration once or twice weekly in a cycle selected from the group consisting of: (i) weekly, (ii) every other week, (iii) 1 week on treatment followed by 2, 3, or 4 weeks rest, (iv) 2 weeks on treatment followed by 1, 2, 3, or 4 weeks rest, (v) 3 weeks on treatment followed by 1, 2, 3, 4, or 5 weeks rest, (vi) 4 weeks on treatment followed by 1, 2, 3, 4, or 5 weeks rest, (vii) 5 weeks on treatment followed by 1, 2, 3, 4, or 5 weeks rest; and (viii) monthly. The cycle may be repeated 4, 6, 8, 10, 12, 16, 20, or more times.

[0096] In some embodiments, the anticancer drug may be administered once every two or three weeks for a total of at least three doses, or twice weekly for four to six weeks. 2 If the dose can be reduced to as low as 340 mg for a 1.7 m patient, or 4.9 mg / kg for a 70 kg patient, it may be administered once or even twice weekly for 4 to 10 weeks. In some embodiments, the dosing schedule may be shortened, i.e., every 2 or 3 weeks for 2 to 3 months. However, higher doses, such as 2 mg / kg once weekly or once every 2 to 3 weeks, may be administered gradually in repeated dosing cycles. It has been determined that administration can be by slow intravenous infusion. The administration schedule can optionally be repeated at other intervals, and the dosage may be administered by various parenteral routes, with appropriate adjustments to the dose and schedule. In some embodiments, the dosage is administered on days 1 and 8 of each 21-day cycle.

[0097] In some embodiments, the methods disclosed herein further comprise administering an antibody-drug conjugate at a dose of about 4 mg / kg to about 12 mg / kg. In some embodiments, the antibody-drug conjugate is administered at a dose of about 8 mg / kg to about 12 mg / kg. In some embodiments, the antibody-drug conjugate is administered at a dose of about 8 mg / kg, about 10 mg / kg, or about 12 mg / kg. In some embodiments, the antibody-drug conjugate is sacituzumab govitecan.

[0098] In some embodiments, the method further comprises one or more additional treatment modalities selected from an antibody, a conjugate, a gene therapy, chemotherapy, radiation therapy, surgery, a BTK inhibitor, and a checkpoint inhibitor.

[0099] In some embodiments, the method further comprises radiation therapy.

[0100] In some embodiments, the anti-cancer agent is selected from a chemotherapeutic agent, a checkpoint inhibitor, a FLT3 agonist, and a BTK inhibitor.

[0101] In some embodiments, the FLT3 inhibitor is GS-3583. FLT3 agonists also include CDX-301 and the agents disclosed in PCT Publication No. 2020 / 263830.

[0102] In some embodiments, the FLT3 agonist is an Fc fusion protein disclosed in WO 2022 / 031876.

[0103] In some embodiments, the disclosure provides a method for treating cancer. The method includes administering an MCL-1 inhibitor and a chemotherapeutic agent for the treatment of cancer; the method further includes administering one or more additional therapeutic agents, with the proviso that the additional therapeutic agents are not FLT3 agonists. In some embodiments, the additional therapeutic agent is not a FLT3-Fc fusion protein. In some embodiments, the antibody-drug conjugate is sacituzumab govitecan, the MCL-1 inhibitor is Compound A, and the additional therapeutic agent is not a FLT3 agonist. In some embodiments, the additional therapeutic agent is not a FLT3 agonist disclosed in WO 2020 / 263830. In some embodiments, the additional therapeutic agent is not a fusion protein comprising the amino acid sequence of SEQ ID NO: 14 of U.S. Patent No. 11 / 124,582.

[0104] In some embodiments, the checkpoint inhibitor is selected from an anti-PD-1 agent, an anti-PD-L1 agent, an anti-PD-1 / PD-L1 interaction inhibitor, an anti-CTLA4 agent, and an anti-TIGIT agent.

[0105] In some embodiments, the checkpoint inhibitor is selected from an anti-PD-1 agent, an anti-PD-L1 agent, an anti-PD-1 / PD-L1 interaction inhibitor, an anti-CTLA4 agent, and an anti-TIGIT agent. In some embodiments, the checkpoint inhibitor is selected from nivolumab, pembrolizumab, atezolizumab, zinbelimab, and pidilizumab. In some embodiments, the checkpoint inhibitor is selected from ipilimumab, lambrolizumab, tremelimumab, durvalumab, avelumab, donbanalimab, and tiragolumab.

[0106] Examples of CTLA4 inhibitors that can be co-administered include ipilimumab, tremelimumab, BMS-986218, AGEN1181, AGEN1884, BMS-986249, MK-1308, REGN-4659, ADU-1604, CS-1002, BCD-145, APL-509, JS-007, BA-3071, ONC-392, AGEN-2041, JHL-1155, KN-044, CG-0161, ATOR-1144, and PBI -5D3H5, BPI-002, and the multispecific inhibitors FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-019 (PD-1 / CTLA4), KN-046 (PD-1 / CTLA4), MEDI-5752 (CTLA4 / PD-1), XmAb-20717 (PD-1 / CTLA4), and AK-104 (CTLA4 / PD-1).

[0107] Examples of PD-L1 (CD274) or PD-1 (PDCD1) inhibitors that can be co-administered include pembrolizumab, nivolumab, cemiplimab, pidilizumab, AMP-224, MEDI0680 (AMP-514), spartalizumab, atezolizumab, avelumab, durvalumab, ALN-PDL, BMS-936559, CK-301, PF-06801591, BGB-108, BGB-A317 (tislelizumab), GLS-010 (WBP-3055), AK-103 (HX-008), GB-226, AK-105, CS-1003, HLX-10, MGA-012, BI-754091, PDR-001, AGEN-2034, JS-001 (toripalimab), JNJ-63723283, genolimuzumab (CBT-501), LZM-009, BCD-100, LY-3300054, SHR-1201, SHR-1210 (camrelizumab), Sym-021, ABBV-181, PD1-PIK, BAT-1306, RO-6084 (PD-L1 antisense oligonucleotide), STI-1110, GX-P2, RG-7446, mDX-400, (MSB0010718C) , CX-072, CBT-502, TSR-042 (dostallimab), MSB-2311, JTX-4014, BGB-A333, SHR-1316, CS-1001 (WBP-3155), MEDI-0680, embafolimab (KN-035), KD-033, KY-1003, IBI-308 (sintilimab), HLX-20, KL-A167, STI-A1014, STI-A1015 (IMC-001), BCD-135, FAZ-053, TQB-2450, MDX1105-01, MSB-0010718C, GS-4224, GS-4 416, INCB086550, MAX10181, and multispecific inhibitors FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-013 (PD-1 / LAG-3), FS-118 (LAG-3 / PD-L1), MGD-019(PD-1 / CTLA4), KN-046(PD-1 / CTLA4), MEDI-5752(CTLA4 / PD-1), RO-7121661(PD-1 / TIM-3), XmAb-20717(PD-1 / CTLA4), AK-104(CTLA4 / PD-1),These include, but are not limited to, M7824 (PD-L1 / TGFβ-EC domain), CA-170 (PD-L1 / VISTA), CDX-527 (CD27 / PD-L1), LY-3415244 (TIM3 / PDL1), GNS-1480 (epidermal growth factor receptor antagonist; programmed cell death ligand 1 inhibitor), M-7824 (PD L1 / TGFβ bifunctional fusion protein), and INBRX-105 (4-1BB / PDL1).

[0108] Examples of PD-1 inhibitors include those disclosed in WO 2017112730 (Incyte Corp), WO 2017087777 (Incyte Corp), WO 2017017624, WO 2014151634 (BristolMyers Squibb Co), WO 201317322 (BristolMyers Squibb Co), WO 2018119286 (Incyte Corp), WO 2018119266 (Incyte Corp), and WO 2018119263 (Incyte Corp). (Same as No. 2018119236 (Incyte Corp), Same as No. 2018119221 (Incyte Corp), Same as No. 2018118848 (BristolMyers Squibb Co), Same as No. 20161266460 (BristolMyers Squibb Co), Same as No. 2017087678 (BristolMyers Squibb Co), Same as No. 2016149351 (BristolMyers Squibb Co), Same as No. 2015033299 (Aurigene Discovery Technologies Ltd), Same as No. 2015179615 (Eisai Co Ltd; Eisai Research Institute), Same as No. 2017066227 (BristolMyers Squibb Co), Same as No. 2016142886 (Aurigene Discovery Technologies) Ltd), Same No. 2016142852 (Aurigene Discovery Technologies Ltd), Same No. 2016142835 (Aurigene Discovery Technologies Ltd; Individual), Same No. 2016142833 (Aurigene Discovery Technologies Ltd., same as No. 2018085750 (BristolMyers Squibb Co.), same as No. 2015033303 (Aurigene Discovery Technologies Ltd.), same as No. 2017205464 (Incyte Corp.), same as No. 2016019232 (3M Co.;Individual;Texas A&M University System), same as No. 2015160641 (BristolMyers Squibb Co.), same as No. 2017079669 (Incyte Corp.). Corp), No. 2015033301 (Aurigene Discovery Technologies Ltd), No. 2015034820 (BristolMyers Squibb Co), No. 2018073754 (Aurigene Discovery Technologies Ltd), No. 2016077518 (BristolMyers Squibb Co), No. 2016057624 (BristolMyers Squibb Co), No. 2018044783 (Incyte Corp), No. 2016100608 (BristolMyers Squibb Co), No. 2016100285 (BristolMyers Squibb Co), No. 2016039749 (BristolMyers Squibb Co), No. 2015019284 (Cambridge Enterprise Ltd), No. 2016142894 (Aurigene Discovery Technologies Ltd), No. 2015134605 (BristolMyers Squibb Co), No. 2018051255 (Aurigene Discovery Technologies Ltd), No. 2018051254 (Aurigene Discovery Technologies Ltd), No. 2017222976 (Incyte Corp), No. 2017070089 (Incyte Corp), No. 2018044963 (BristolMyers Squibb Co), No. 2013144704 (Aurigene Discovery Technologies Ltd), No. 2018013789 (Incyte Corp), No. 2017176608 (BristolMyers Squibb Co), No. 2018009505 (BristolMyers Squibb Co), No. 2011161699 (Aurigene Discovery Technologies Ltd), No. 2015119944 (Incyte Corp;Merck Sharp & Dohme Corp), Merck No. 2017192961 (Incyte Corp), Merck No. 2017106634 (Incyte Corp), Merck No. 2013132317 (Aurigene Discovery Technologies Ltd), Merck No. 2012168944 (Aurigene Discovery Technologies Ltd), Merck No. 2015036927 (Aurigene; Discovery Technologies Ltd), No. 2015044900 (Aurigene Discovery Technologies Ltd), and compounds described in Arising International, No. 2018026971.

[0109] The PD-1 / PD-L1 inhibitor can be administered in any suitable amount known to one of skill in the art. In some embodiments, the compound of Formula I is administered to a subject in an amount of 0.1 to 1000 mg. Representative amounts of the PD-1 / PD-L1 inhibitor administered to a subject include, but are not limited to, 0.1 to 500 mg, 1 to 100 mg, 1 to 50 mg, or 10 to 50 mg. Other amounts of the PD-1 / PD-L1 inhibitor administered to a subject include, but are not limited to, about 1 mg, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or about 100 mg.

[0110] In some embodiments, the methods described herein further comprise administering an anti-TIGIT antibody, such as BMS-986207, RG-6058, or AGEN-1307.

[0111] In some embodiments, the methods described herein further comprise administering a BTK (Bruton's tyrosine kinase) inhibitor. Examples of such BTK inhibitors are the compounds disclosed in U.S. Patent No. 7,405,295. Additional examples of BTK inhibitors include, but are not limited to, (S)-6-amino-9-(1-(but-2-ynoyl)pyrrolidin-3-yl)-7-(4-phenoxyphenyl)-7H-purin-8(9H)-one, acalabrutinib (ACP-196), BGB-3111, HM71224, ibrutinib, M-2951, tirabrutinib (ONO-4059), PRN-1008, spebrutinib (CC-292), and TAK-020. In some embodiments, the BTK inhibitor is selected from acalabrutinib, tirabrutinib, zanubrutinib, and PCI-32765.

[0112] In some embodiments, the anti-cancer agent is a chemotherapeutic agent.

[0113] In some embodiments, the anti-cancer agent is selected from the agents of Example 3 listed in Table 21. In some embodiments, the chemotherapeutic agent is docetaxel. In some embodiments, the chemotherapeutic agent is gemcitabine. In some embodiments, the chemotherapeutic agent is paclitaxel.

[0114] In some embodiments, the anticancer drug is doxorubicin (DOX), Pyrubicin, morpholinodoxorubicin (morpholino-DOX), cyanomorpholino-doxorubicin (cyanomorpholino-DOX), 2-pyrrolino-doxorubicin (2-PDOX), CPT, 10-hydroxycamptothecin, SN- 38, topotecan, lutotecan, 9-aminocamptothecin, 9-nitrocamptothecin, taxanes, geldanmycin, ansamycins, and epothilones.

[0115] In some embodiments, the methods disclosed herein further comprise administering an antibody-drug conjugate. In some embodiments, the antibody-drug conjugate comprises an anti-Trop-2 antibody. In some embodiments, the antibody-drug conjugate is sacituzumab govitecan. In some embodiments, the antibody-drug conjugate is datopotamab deruxtecan.

[0116] In some embodiments, when the agents of the present disclosure are combined with one or more additional therapeutic agents described herein, the components of the composition are administered simultaneously or as a sequential regimen. When administered sequentially, the combination may be administered in two or more doses. In some embodiments, the agents of the present disclosure When an agent is combined with one or more additional therapeutic agents described herein, the components of the composition are administered simultaneously or as a sequential regimen. When administered sequentially, the combination may be administered in two or more doses. In certain embodiments, when an agent of the present disclosure is combined with one or more additional therapeutic agents described herein, the components of the composition are administered simultaneously or as a sequential regimen. When administered sequentially, the combination may be administered in two or more doses.

[0117] Co-administration of an agent disclosed herein with one or more additional therapeutic agents generally refers to the simultaneous or sequential administration of an agent disclosed herein with one or more additional therapeutic agents such that a therapeutically effective amount of each agent is present in the patient's body.

[0118] Co-administration includes administration of a unit dose of an agent disclosed herein before or after administration of a unit dose of one or more additional therapeutic agents. The agents disclosed herein can be administered within seconds, minutes, or hours of administration of the one or more additional therapeutic agents. For example, in some embodiments, a unit dose of an agent disclosed herein is administered first, followed within seconds or minutes by a unit dose of one or more additional therapeutic agents. Alternatively, in other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed within seconds or minutes by a unit dose of an agent disclosed herein. In some embodiments, a unit dose of an agent disclosed herein is administered first, followed several hours (e.g., 1-12 hours) later by a unit dose of one or more additional therapeutic agents. In other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed several hours (e.g., 1-12 hours) later by a unit dose of an agent disclosed herein.

[0119] In some embodiments, the disclosure provides methods for treating or preventing cancer. In certain embodiments, the disclosure provides methods for treating or preventing cancer, comprising administering to an individual a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer is a hematological cancer. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is selected from the group consisting of breast cancer, colorectal cancer, skin cancer, melanoma, ovarian cancer, renal cancer, small cell lung cancer, non-small cell lung cancer, lymphoma, and leukemia. [Example]

[0120] Example 1: In vitro synergy of MCL-1 inhibitors with SN-38 in TNBC and NSCLC cell lines To test the potential for combination between Compound A and SN-38 (a topoisomerase inhibitor), in vitro studies were conducted in a panel of triple-negative breast cancer (TNBC, n=3) and non-small cell lung cancer (NSCLC, n=2) cell lines using the Bliss independence model of synergy. Cells were exposed to a dose-titrated matrix of each compound alone and in combination for 72 hours, and cell viability was then determined by Cell Titer Glo reagent. Strong Bliss synergy scores (>100) were observed in all cell lines tested. Materials and Methods Cell culture and reagents

[0121] HCC70 (ATCC® CRL-2315), HCC1806 (ATCC® CRL-2335), HCC1187 (ATCC® CRL-2322), NCI-H522 (ATCC® CRL-5810), and H820 (ATCC® HTB-181) cell lines were thawed from liquid nitrogen storage and maintained in RPMI-1640 (Gibco-12633) + 10% HI-FBI (Gibco-16140) + Pen / Strep (100x Gibco-15140) according to ATCC guidelines. Cells were passaged using 0.25% trypsin / EDTA (1x GIBCO-25200) according to ATCC guidelines.

[0122] SN-38 and Compound A stocks (provided by the Gilead sample bank) were dispensed directly into treatment wells using a D300e Digital Dispenser (vendor) with DMSO (Sigma-D2438) to 0.1% v / v as a vehicle control.

[0123] Viability assessment was performed using Cell Titer Glo™ (Promega #G9241) according to the manufacturer's microwell plate protocol and read for luminescence on a Biotek Synergy Neo2 plate reader. Cell viability combination assay

[0124] For the synergy matrix assay, cell lines were seeded at 5,000 cells per well in clear-bottom white 96-well plates (Corning #3909) in 100 μL of the recommended cell culture medium. Treatment maps consisted of a single-agent dose response for Compound A (seven 3-fold dilutions + no-treatment control) or SN-38 (nine 3-fold dilutions + no-treatment control) and a checkerboard matrix of 63 different combinations. Concentration ranges were selected based on the relative sensitivity of each cell line to the compounds. Five plates were used for each combination to generate sufficient replicates to calculate synergy scores with 95% confidence intervals (95% CI).

[0125] Compounds and DMSO vehicle were applied to cells using an HP D300 dispenser and dispensed directly into the medium according to a checkerboard matrix, and viability was measured by Cell Titer Glo after 72 hours of incubation at 37°C / 5% CO2 / 100% relative humidity. Data analysis

[0126] Combination viability data were evaluated for synergy using the Excel template described by Prichard and Shipman {Prichard 1990}. Specifically, the single-component dose curves for SN-38 and Compound A were normalized to the percent viability on each plate and averaged across five technical replicates to calculate the theoretical additive kill of the combination according to the Bliss principle of independence. Calculated values ​​were compared with experimental results generated using a 63-concentration checkerboard. Synergy or antagonism scores were generated depending on whether the observed growth inhibition was greater or less than the calculated value, respectively.

[0127] For example, if two compounds (B) and (C) at given concentrations each produce 60% inhibition, their theoretical additive inhibition will be 84% according to the Bliss independence formula: 60% B +60% C * (100%-60% B )=84% B+C If the experimental result is greater than the calculated value (e.g., 90% inhibition), the difference [6%] is added to the synergy score. If the result is less (e.g., 78% inhibition), the difference [6%] is added to the antagonism score.

[0128] These differences were summed across the entire checkerboard (63 wells) to obtain cumulative synergy and antagonism scores in μM 2 The scores were given in %, reflecting a 2D surface of the dose-response. A 95% confidence interval adjustment was applied to the synergy and antagonism scores, and each sum was compared to the original scale: a score above 50 was considered moderate synergy, and a score above 100 was considered strong synergy, likely indicating a combined effect in vivo. {Prichard 1990}.

[0129] Data from combination assays are presented in three formats. Synergy scores with 95% confidence intervals were averaged from n=2 assays. Exemplary tabular and graphical formats for each cell line. Percent inhibition matrix of Example tabular and graphical synergy matrices with 95% confidence intervals for each cell line.

[0130] To test the potential for combination between Compound A and SN-38 (a topoisomerase inhibitor), in vitro studies were conducted in a panel of TNBC (n=3) and NSCLC (n=2) cell lines using the Bliss independence model of synergy. Cells were exposed to a dose-titrated matrix of each compound alone and in combination for 72 hours, and cell viability was then determined using Cell Titer Glo reagent. Strong Bliss synergy scores (>100) were observed in all cell lines tested. [Table 1] [Table 2] [Table 3] [Table 4] [Table 5] [Table 6] [Table 7] [Table 8] [Table 9] [Table 10] [Table 11] [Table 12] * The SN-38 dose response was shifted to a maximum concentration of 1.0 μM, capturing a large potential range of synergistic activity. Example 2: In vitro synergy of MCL-1 inhibitors with paclitaxel in TNBC cells Materials and Methods Cell culture and reagents

[0131] HCC70 (ATCC® CRL-2315) and HCC1806 (ATCC® CRL-2335) were thawed from liquid nitrogen storage and maintained in RPMI-1640 (Gibco-12633) + 10% HI-FBI (Gibco-16140) + Pen / Strep (100x Gibco-15140) according to ATCC guidelines. MDA-MB-468 (ATCC® HTB-132) cells were thawed and maintained in DMEM (Gibco-11995) + 10% HI-FBS + Pen / Strep. Cells were passaged according to ATCC guidelines using 0.25% trypsin / EDTA (1x GIBCO-25200).

[0132] Paclitaxel) and Compound A stock (provided by the Gilead sample bank) were dispensed directly into treatment wells using a D300e Digital Dispenser (vendor) with DMSO (Sigma-D2438) to 0.1% v / v as a vehicle control.

[0133] Viability assessment was performed using Cell Titer Glo™ (Promega #G9241) according to the manufacturer's microwell plate protocol and analyzed using Synergy Luminescence was read on a Neo2 plate reader.

[0134] Cell lysates for the MSD assay were generated using 1x lysis buffer (10x Cell Signaling CST-9803), 100x protease inhibitors, phosphatase inhibitor I, phosphatase inhibitor II (Meso Scale Discovery Inhibitor Pack R70AA-1), and PMSF (SIGMA catalog no. 7626).

[0135] MCL-BAK and MCL1-BIM dimer assays and total MCL1 assays were developed by MSD Custom Assay Services and performed using the MSD U-PLEX Development Pack (K15227N) and protocol revision "2018Mar rev 2." GAPDH was determined by MSD using standard assay K151PWD. All plates were read on an MSD SECTOR Imager 2400 using MSD Read Buffer T (R92TC).

[0136] Protein Simple Reagents: EZ Standard Pack 1 (PS-ST01EZ: Biotinylated Ladder, FL Standard, and DTT), Peroxide (044-379), Luminal-S (043-311), Antibody Dilution Buffer (042-203), Streptavidin-HRP (042-414), Secondary Antibodies: Goat Anti-Rabbit (042-206) and Goat Anti-Mouse (042-205), Separation Matrix (042-512), Stacking Matrix (042-513), 10X Sample Buffer (042-195), Wash Buffer (042-520), Upper Running Buffer (043-163), Lower Running Buffer (043-162), 384-well Plate (040-663), Sizing Capillary (55700), Protein Simple Instruments Peggy Sue™ and Sally Sue(trademark). Primary antibodies: MCL1 (CST-94296), FBXW7 (Abcam 109617 and Abcam 171961). Cell viability combination assay

[0137] For the synergy matrix assay, TNBC cell lines were seeded at 10,000 cells per well in clear-bottom white 96-well plates (Corning #3909) in 100 μL of the recommended cell culture medium. Plates were incubated at 37°C and 100% RH for 20 h before compound exposure. Treatment maps consisted of single-agent dose responses of Compound A (seven 3-fold dilutions from 3 μM to 4 nM + no-treatment control) or paclitaxel (nine 3-fold dilutions from 3 μM to 0.5 nM + no-treatment control) and a checkerboard matrix of 63 different combinations. Five plates were used for each combination to generate sufficient replicates to calculate synergy scores with 95% confidence intervals (95% CI).

[0138] Paclitaxel and DMSO vehicle were first applied to cells using an HP D300 dispenser and aliquoted directly into the medium according to a checkerboard matrix. Paclitaxel was incubated for 4 hours, then washed off by removing the medium, washing twice with 200 μL of prewarmed complete medium, and finally replacing with 100 μL of prewarmed complete medium. Cells were then exposed to Compound A in the same checkerboard matrix using the D300 dispenser and incubated for 48 hours before measuring viability by Cell Titer Glo. MSD assay

[0139] For the MSD assay, TNBC cell lines were seeded at 25,000 cells per well in clear-bottom white 96-well plates (Corning #3909) in the recommended cell culture medium. Plates were incubated at 37°C and 100% RH for 20 hours before compound exposure. Paclitaxel and DMSO vehicle were first applied to the cells using an HP D300 dispenser and aliquoted directly into the medium. Paclitaxel was incubated for 4 hours, then washed away by removing the medium, washing twice with 150 μL of prewarmed complete medium, and finally replacing with 100 μL of prewarmed complete medium. After an additional 20 hours, samples were collected by aspirating the supernatant, and 125 μL of 1× lysis buffer was added to each well. Plates were briefly placed on ice and transferred to a rocking platform at 4°C for 20 minutes. Plates were placed on dry ice, flash-frozen for 10 minutes, and then stored at -80°C until testing.

[0140] MCL1 and MCL1-BAK and MCL1-BIM dimer assays were performed using materials and protocols provided by MSD Custom Assay Services based on their U-Plex technology. Plates were first prepared using the standard U-PLEX capture antibody coating protocol and then washed three times with 150 μL of MSD wash buffer. 25 μl of sample or standard was added directly to the plate, which was then sealed and incubated at room temperature for 1 hour with shaking. The plate was washed three times with 150 μL of wash buffer per well, and 50 μL of antibody detection solution was added to each well of the MSD plate. The plate was sealed and incubated at room temperature for 1 hour with shaking. The plate was again washed three times with 150 μL of wash buffer per well. 150 μL of 2x read buffer was added to each well, and the plate was read on an MSD SECTOR Imager 2400.

[0141] The GAPDH assay kit (MSD) was run according to the manufacturer's protocol, using 25 μL of lysate per sample added directly to the plate. The plate was sealed and incubated at room temperature with shaking for 1 hour, then washed three times with 150 μL of wash buffer per well. 25 μL of antibody detection solution was added to each well, the plate was sealed and incubated at room temperature with shaking for 1 hour, then washed three times with 150 μL of wash buffer per well. 150 μL of 2× read buffer was then added to each well, and the plate was measured for electrochemiluminescence (ECL) on an MSD SECTOR Imager 2400. Simple Western

[0142] Simple Western immunoassays are performed in capillaries. Samples and reagents are loaded into an assay plate and placed in the Protein Simple Instrument. Cell lysates are automatically loaded into the capillaries and separated by size as they migrate through a stacking and separation matrix. Separated proteins are then immobilized to the capillary wall via a proprietary light-activated capture chemistry. Target proteins are identified using a primary antibody and immunoprobed with an HRP-conjugated secondary antibody and a chemiluminescent substrate. The resulting chemiluminescent signal is detected and quantified.

[0143] FBXW7 expression was measured in cell lines after 4 hours of treatment with 1 μM paclitaxel, followed by washing and overnight incubation using Simple Western. Lysates were prepared and diluted to 0.5 μg / ml in 1× lysis buffer. The Simple Western platform is run on a 384-well plate.

[0144] The marker, internal ladder, and DTT are provided in lyophilized form by Simple Western. Resuspend the reagents as described in the protocol. Add 20 μL of water to the marker. Add 40 μL of water to the DTT, and mix 20 μL of 10X sample buffer and 20 μL of DTT. This is referred to as Z buffer. Load the marker into well 1A. Add 5 μL of lysate to a 1.7 mL Eppendorf tube. Add 1.2 μL of Z reagent to each sample. Heat the samples at 100°C for 5 minutes, cool, and then spin in a microcentrifuge for 30 seconds. Load the samples into wells A2-12. Dilute the primary antibody 1:50 in antibody dilution buffer (6 μL + 294 μL dilution buffer). Load 20 μL of each antibody into lanes 2-12. Each row can contain up to eight different antibodies. Actin diluted 1:300 was used as a loading control. Further loading of either goat anti-rabbit or goat anti-mouse secondary antibody is performed as needed. The plate is rotated at 2.6k for 10 minutes at room temperature, loaded onto the instrument, and run overnight. Target proteins are identified using primary antibodies and immunoprobed using HRP-conjugated secondary antibodies and a chemiluminescent substrate. The resulting chemiluminescent signal is detected and quantified. Data analysis About Bliss Synergy

[0145] Combination viability data were evaluated for synergy using the Excel template described by Prichard and Shipman {Prichard 1990}. Specifically, the single-component dose curves for paclitaxel and Compound A were normalized to the percent viability on each plate and averaged across five technical replicates to calculate the theoretical additive kill of the combination according to the Bliss principle of independence. Calculated values ​​were compared to experimental results generated with a 63-concentration checkerboard. Synergy or antagonism scores were generated depending on whether the observed growth inhibition was greater or less than the calculated value, respectively.

[0146] For example, if two compounds (B) and (C) at given concentrations each produce 60% inhibition, their theoretical additive inhibition will be 84% according to the Bliss independence formula: 60% B +60% C * (100%-60% B )=84% B+C

[0147] If the experimental result is greater than the calculated value (e.g., 90% inhibition), the difference [6%] is added to the synergy score. If the result is less (e.g., 78% inhibition), the difference [6%] is added to the antagonism score.

[0148] These differences were summed across the entire checkerboard (63 wells) to obtain cumulative synergy and antagonism scores in μM 2 The scores were given in %, reflecting a 2D surface of the dose-response. A 95% confidence interval adjustment was applied to the synergy and antagonism scores, and each sum was compared to the original scale: a score above 50 was considered moderate synergy, and a score above 100 was considered strong synergy, likely indicating a combined effect in vivo. {Prichard 1990}.

[0149] Data from combination assays are presented in three formats: Synergy scores with 95% confidence intervals averaged from n=2 assays; Exemplary percent inhibition matrix in tabular and graphical form for each cell line; Exemplary synergy matrix in tabular and graphical form with 95% confidence intervals for each cell line. MSD assay

[0150] For total MCL1, ECL signals were recorded and converted to pg / mL using calibration controls developed at MSD over an eight-point standard dose range (0-10,000 pg / mL) using a four-parameter curve-fitting function in MSD WorkBench software. Results for MCL1-BAK and MCL1-BIM dimers were converted to pg / mL using the same process, except standard concentrations ranged from 0-50,000 pg / mL. GAPDH results were recorded and reported as ECL and used to normalize both MCL1 and MCL1 dimer within each sample set. For graphical comparisons between analytes for a given cell line, each pg / mL data set was normalized to the vehicle control at 100%, with no protein at 0%.

[0151] Paclitaxel has been reported to downregulate MCL1 protein levels, in part through an increase in the MCL1 E3-ligase FBXW7, which targets MCL1 for proteasomal degradation {Wertz 2011}. To confirm this observation, HCC70, MDA-MB-468, and HCC1806 TNBC cell lines were treated with a clinically relevant concentration of paclitaxel (1 μM) for 4 hours (Gianni 1995). After paclitaxel treatment, cells were incubated overnight, and protein levels were determined. Comparison of paclitaxel treatment with vehicle control revealed increased FBXW7 protein levels and decreased MCL1 protein levels (Table 13 and Figure 1). Paclitaxel treatment also resulted in decreased protein levels of MCL1-BAK and MCL1-BIM dimers (Table 13 and Figure 1). These results were observed across all three TNBC cell lines (n = 3 biological replicates).

[0152] HCC70, MDA-MB-468, and HCC1806 TNBC cell lines were treated with a clinically relevant concentration of paclitaxel (1 μM) for 4 hours (Gianni 1995). Paclitaxel treatment increased FBXW7 protein levels, decreased MCL1 protein levels, and decreased MCL1-BAK and MCL1-BIM dimer protein levels in all three cell lines. HCC70, MDA-MB-468, and HCC1806 were treated with paclitaxel dose-escalation (protein-adjusted C). max Bliss synergy (>100) was observed when mice were pretreated with 100 μM of Compound A (including 1 μM) for 4 hours to mimic clinical exposure, followed by 72 hours of exposure to increasing doses of Compound A. [Table 13] a Mean normalized protein levels (n = 3 biological replicates)

[0153] Bliss synergy was used to determine whether the reduction in MCL1 protein levels after paclitaxel treatment resulted in enhanced sensitivity to Compound A. HCC70, MDA-MB-468, and HCC1806 were treated with paclitaxel dose escalation (protein-adjusted C). max = 1 μM) for 4 hours to mimic clinical exposure and then exposed to increasing doses of Compound A. Cells were incubated for 72 hours and viability was determined using the CTG reagent. Bliss synergy was observed across all three TNBC cell lines exposed to the combination of Compound A and paclitaxel in vitro (Table 15). A Bliss synergy score of greater than 100 is considered a strong effect {Prichard 1990}. [Table 14] [Table 15] [Table 16] [Table 17] [Table 18] [Table 19] [Table 20] Example 3: The potential combination of Compound A with targeted agents and chemotherapy was tested in a panel of breast cancer cell lines using a 72-hour in vitro proliferation assay. The results of the combination studies are shown in Table 21. [Table 21-1] [Table 21-2] [Table 21-3] [Table 21-4] [Table 21-5] [Table 21-6] [Table 21-7] [Table 21-8] [Table 21-9] [Table 21-10] [Table 21-11] [Table 21-12] [Table 21-13] [Table 21-14] [Table 21-15] [Table 21-16] [Table 21-17] [Table 21-18] Example 4: The potential combination of Compound A with BTK inhibitors was tested in a panel of hematological cancer cell lines using a 72-hour in vitro proliferation assay. The results of the combination study are shown in Table 22. [Table 22] Example 5: Combination potential of Compound A and SN-38 was assessed using a 72-hour in vitro proliferation assay The combination assay was used to test a panel of TNBC and NSCLC cancer cell lines. The results of the combination studies are shown in Table 23. [Table 23] Example 6: The potential combination of Compound A and paclitaxel was tested in an in vivo TNBC PDX model in an antitumor efficacy study (Figures 5 and 6). Example 7: Phase 1a / b Study to Evaluate the Safety, Tolerability, and Pharmacokinetics of Compound A as Monotherapy and in Combination with Anticancer Therapy in Subjects with Solid Malignancies

[0154] The study will be conducted to characterize the safety and tolerability of Compound A and Compound A in combination with anti-cancer therapy in subjects with advanced solid malignancies. Test Design

[0155] This is an open-label, multicenter, dose-escalation, and dose-expansion Phase 1a / 1b study to evaluate the safety, tolerability, and PK profile of Compound A, document any DLTs (dose-limiting toxicities), and determine the MTD (maximum tolerated dose) and / or RP2D (recommended phase 2 dose) of Compound A as monotherapy and in combination with anticancer therapy in subjects with advanced solid malignancies. The RP2D is the dose level(s) with acceptable tolerability, exposure, efficacy, and biomarker activity. The study consists of two phases: Phase 1a (dose expansion), followed by Phase 1b (dose expansion). Phase 1a Dose Escalation: Part A: Dose Escalation of Compound A as Monotherapy Phase 1b dose expansion: Part B: Optional disease-specific cohorts of Compound A in combination with anti-cancer therapies in parallel with Part A Part C: Safety run-in and expansion of Compound A in combination with anti-cancer therapies following Parts A and B.

[0156] Each part of the study consists of a screening, treatment, and follow-up period. Screening will occur up to 28 days before the first dose of study treatment, during which subject eligibility and baseline characteristics will be determined. Part A: Phase 1a dose escalation of Compound A as monotherapy.

[0157] Subjects with advanced solid tumors who have failed or intolerant to standard therapy, or for which no standard therapy exists, will be sequentially enrolled to receive Compound A as monotherapy at increasingly higher dose levels.

[0158] Dose escalation will be performed using a dose escalation design based on the 3+3 rule.

[0159] Compound A will be administered orally on days 1, 2, 8, 9, 15, and 16 of each 21-day cycle for up to 105 weeks.

[0160] Up to six cohorts (i.e., six dose levels) of 3-6 subjects each will receive escalating dose levels of Compound A as monotherapy. The planned starting dose of Compound A is 5 mg, with target doses of 15 mg and 50 mg for the following two cohorts. Subsequent dose levels following the starting dose will be determined based on safety, tolerability, and PK (pharmacokinetics) data from previous cohorts. The dose level increase will be determined based on all available clinical data, including the clinical status of the patient, and approved by the Safety Review Team (SRT), potentially up to 300 mg. The increase in dose level will be no more than half a logarithm with each subsequent dose escalation.

[0161] The safety and tolerability of each dose level will be assessed by the SRT after all subjects within a cohort have been followed for at least 21 days after the first dose of Compound A or after a subject has a DLT during the first 21 days of study drug administration.

[0162] The first block of each dose consists of three subjects. If no subjects experience a DLT during the first 21 days of study drug administration, dose escalation will occur. If one subject in the initial cohort of three subjects experiences a DLT during the first 21 days of study drug administration, three additional subjects will be enrolled at the same dose level. If no DLT is observed in the three additional subjects, dose escalation will occur. If two or more subjects experience a DLT within the first 21 days, dose de-escalation to a lower dose will occur. The MTD is the highest dose level with a subject incidence of DLT of less than 33% during the first 21 days of study drug administration.

[0163] 21 days of treatment with a consistent regimen in any given cohort for the decision rule to apply.

[0164] Throughout the study, subjects with malignant tumors for which a biopsy is available may undergo optional tumor biopsies. These subjects must consent and provide separate, specific written consent. Dose escalation criteria

[0165] For any given cohort, the sponsor may choose to withhold dosing, select an intermediate dose, or stop study enrollment at any time based on a review of preliminary safety and available PK and / or pharmacodynamic data.

[0166] Based on review of relevant safety and available PK and / or pharmacodynamic data by the SRT, escalation to higher dose cohorts will occur only in the absence of DLTs and / or meeting any pre-specified stopping criteria. Dose escalation to subsequent cohorts on a greater than half-log scale requires affirmation by at least two-thirds of the SRT.

[0167] Dose-limiting toxicities are defined as the following Compound A-related events occurring within the first 21 days (after the first dose of Compound A): Grade 4 hematologic toxicity lasting more than 21 days. All Compound A-related Grade 3 non-hematologic toxicities lasting more than 7 days and all Compound A-related Grade 4 non-hematologic toxicities of any duration will be considered DLTs. Part B: Optional disease-specific cohorts of compound A in combination with anti-cancer therapy in parallel with Part A

[0168] During monotherapy dose escalation in Part A and prior to the formal dose expansion of sponsor-nominated and supported disease-specific cohorts featuring combination therapy with Compound A in Part C, the sponsor may choose to nominate and support one or more of the following cohorts aligned with those in Part C for combination therapy with Compound A at any previously evaluated dose in Part A that was deemed safe and tolerable by the SRT: Cohort B1: Metastatic NSCLC (Compound A + docetaxel) Cohort B2: Metastatic NSCLC (compound A + sacituzumab govitecan) Cohort B3: Metastatic TNBC (Compound A + docetaxel) Cohort B4: Metastatic TNBC (compound A + sacituzumab govitecan) Cohort B5: mSTS with non-specific histology (Compound A + docetaxel and gemcitabine)

[0169] Each additional cohort consists of a single such population with a particular combination. Part C: Safety run-in and dose expansion of Compound A in combination with other anticancer therapies

[0170] This is an open-label Phase 1b study with Compound A given in combination with one or more other anticancer therapies in the following five disease-specific cohorts designated and supported by the sponsor after completion of Parts A and B: Cohort C1: Metastatic NSCLC (Compound A + docetaxel) Cohort C2: Metastatic NSCLC (compound A + sacituzumab govitecan) Cohort C3: Metastatic TNBC (Compound A + docetaxel) Cohort C4: Metastatic TNBC (compound A + sacituzumab govitecan) Cohort C5: mSTS with non-specific histology (Compound A + docetaxel and gemcitabine)

[0171] The RP2D is the dose level(s) with acceptable tolerability, exposure, and biomarker activity.

[0172] The SRT will recommend an initial dose of Compound A for use in combination for each cohort based on the totality of clinical, safety, PK, and pharmacodynamic data. A safety run-in group of at least 3 subjects and no more than 6 subjects will be enrolled to ensure the combination is safe and tolerable in each subject population.

[0173] The safety run-in will employ the same 3+3 design and dose escalation rules as Part A, and will use the same DLT criteria and DLT assessment window as Part A to determine the MTD and / or RP2D. A minimum of 6 subjects must be treated at a dose level before this dose level can be expanded. If cognate disease-specific cohorts from Part B are studied in the RP2D, those subjects may be counted and considered toward the safety run-in group requirements.

[0174] The expansion will include approximately 30 subjects less any disease-specific cognate subjects treated with the same regimens studied in combination under Part B and in any safety run-in. For each disease-specific cohort (B1+C1, B2+C2, B3+C3, B4+C4, B5+C5), a minimum of 20 subjects in RP2D will be enrolled, including any subjects from Part B and / or safety run-in. Cohort C1: Compound A in combination with docetaxel in metastatic NSCLC after single-line therapy for metastatic disease

[0175] Cohort C1 will evaluate the safety and tolerability and define the DLT(s) and MTD and / or RP2D of Compound A in combination with docetaxel in subjects with metastatic NSCLC after a single line of therapy for metastatic disease.

[0176] Compound A will be administered on days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0177] Docetaxel was administered to subjects with an acceptable neutrophil count on the day of administration, specifically ≥ 1500 cells / mm 3 75 mg / m administered as an IV infusion over 1 hour on day 1 of every 21-day cycle, provided 2 The medication is administered at a body surface area (BSA).

[0178] Treatment will continue for up to 105 weeks unless one or more discontinuation criteria are met. Cohort C2: Survival in metastatic NSCLC after single-line therapy for metastatic disease Compound A in combination with situzumab govitecan

[0179] Cohort C2 will evaluate the safety and tolerability and define the DLT(s) and MTD and / or RP2D of Compound A in combination with sacituzumab govitecan in subjects with metastatic NSCLC after a single line of therapy for metastatic disease.

[0180] Compound A will be administered on days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0181] Sacituzumab govitecan is administered to patients with an acceptable neutrophil count on the day of administration, specifically, ≥ 1500 cells / mm on Day 1 of any cycle. 3 or absolute neutrophil count (ANC) ≥ 1000 cells / mm on day 8 of any cycle 3 10 mg / kg is administered as an IV infusion once weekly on days 1 and 8 of every 21-day cycle, provided that: The first infusion should be administered over 3 hours, with the subject being observed both during and for at least 30 minutes after the infusion for signs or symptoms of an infusion-related reaction. Subsequent infusions, if the previous infusion was tolerated, should be administered over 1-2 hours, with the subject being observed both during and for at least 30 minutes after the infusion.

[0182] Treatment will continue for up to 105 weeks unless one or more discontinuation criteria are met. Cohort C3: Compound A in combination with docetaxel in metastatic TNBC after single line therapy for metastatic disease.

[0183] Cohort C3 will evaluate the safety and tolerability and define the DLT(s) and MTD and / or RP2D of Compound A in combination with docetaxel in subjects with metastatic TNBC after a single line of therapy for metastatic disease.

[0184] Compound A will be administered on days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0185] Docetaxel was administered to subjects with an acceptable neutrophil count on the day of administration, specifically ≥ 1500 cells / mm 3 75 mg / m administered as an IV infusion over 1 hour on day 1 of every 21-day cycle, provided 2 The drug is administered with BSA.

[0186] Treatment will continue for up to 105 weeks unless one or more discontinuation criteria are met. Cohort C4: Sacituzumab govitecan plus Compound A in metastatic TNBC after single-line therapy for metastatic disease

[0187] Cohort C4 will evaluate the safety and tolerability and define the DLT(s) and MTD and / or RP2D of Compound A in combination with sacituzumab govitecan in subjects with metastatic TNBC after a single line of therapy for metastatic disease.

[0188] Compound A will be administered on days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0189] Sacituzumab govitecan is administered to patients with an acceptable neutrophil count on the day of administration, specifically, ≥ 1500 cells / mm on Day 1 of any cycle. 3 or ANC ≥ 1000 cells / mm on day 8 of any cycle 3 10 mg / kg is administered as an IV infusion once weekly on days 1 and 8 of every 21-day cycle, provided that: The first infusion should be administered over 3 hours, with the subject being observed both during and for at least 30 minutes after the infusion for signs or symptoms of an infusion-related reaction. Subsequent infusions, if the previous infusion was tolerated, should be administered over 1-2 hours, with the subject being observed both during and for at least 30 minutes after the infusion.

[0190] Treatment will continue for up to 105 weeks unless one or more discontinuation criteria are met. Cohort C5: Metastatic soft tissue sarcoma with nonspecific histology not previously treated for metastatic disease.

[0191] Cohort C5 will evaluate the safety and tolerability and define the DLT(s) and MTD and / or RP2D of Compound A in combination with gemcitabine and docetaxel in subjects with previously untreated soft tissue sarcoma.

[0192] Compound A will be administered on days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0193] Gemcitabine will be administered at a fixed dose rate of 900 mg / m2 BSA as an IV infusion over 90 minutes on days 1 and 8, and docetaxel will be administered at 100 mg / m2 BSA IV over 60 minutes on day 8 of every 21-day cycle.

[0194] Treatment will continue for up to 105 weeks unless one or more discontinuation criteria are met. Duration of treatment

[0195] Study Drug Compound A will be administered for up to 105 weeks or until disease progression, unacceptable toxicity, substantial non-compliance with study procedures or study drug, study discontinuation, withdrawal from the study, or other reason, whichever occurs first. References Ashkenazi A, Fairbrother WJ, Leverson JD, Souers AJ. From basic apoptosis discoveries to advanced selective BCL-2 family inhibitors. Nat Rev Drug Discov 2017;16(4):273-84. Gianni L,Kearns CM,Giani A,Capri G,Vigano L,Lacatelli A,et al.Nonlinear pharmacokinetics and metabolism of paclitaxel and its pharmacokinetic / pharmacodynamic relationships in humans.J Clin Oncol 1995;13(1):180-90. Juin P,Geneste O,Gautier F,Depil S,Campone M.Decoding and unlocking the BCL-2 dependency of cancer cells.Nat Rev Cancer 2013;13(7):455-65. Prichard MN,Shipman C,Jr.A three-dimensional model to analyze drug-drug interactions.Antiviral Res 1990;14(4-5):181-205. Ruefli-Brasse A,Reed JC.Therapeutics targeting Bcl-2 in hematological malignancies.Biochem J 2017;474(21):3643-57. Wertz IE,Kusam S,Lam C,Okamoto T,Sandoval W,Anderson DJ,et al.Sensitivity to antitubulin chemotherapeutics is regulated by MCL1 and FBW7.Nature 2011;471(7336):110-4. Youle RJ,Strasser A.The BCL-2 protein family:opposing activities that mediate c ell death.Nat Rev Mol Cell Biol 2008;9(1):47-59.

[0196] According to a preferred embodiment of the present invention, for example, the following is provided: (Section 1) 1. A method of treating cancer, comprising: administering to a human patient in need thereof a therapeutically effective amount of an anti-cancer agent and a therapeutically effective amount of an MCL-1 inhibitor; The MCL-1 inhibitor is of formula (I):

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Claims

[Claim 1] The invention described in the specification.

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