Wee1 inhibitor combination therapy

Abstract

Disclosed herein is use of a combination of azenosertib or a pharmaceutically acceptable salt thereof and a vedotin-containing antibody-drug conjugate for treating a cancer.

Description

ZENO.191 WO PATENT WEE1 INHIBITOR COMBINATION THERAPYINCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS

[0001] Any and all applications for which a foreign or domestic priority claim is identified in the Application Data Sheet as filed with the present application are hereby expressly incorporated by reference under 37 CFR 1.57, and Rules 4.18 and 20.6, including U.S. Provisional Application No. 63 / 763,064, filed February 25, 2025, the contents of which is hereby incorporated by reference in its entirety including any drawings.SEQUENCE STATEMENT

[0002] This application contains a Sequence Listing, which has been submitted electronically and is hereby incorporated by reference in its entirety. The sequence listing, was created on February 23, 2026, is named SequenceListing_ZENO191WO.xml and is 23,520 bytes in size.Field

[0003] The present application relates to the fields of chemistry, biochemistry and medicine. More particularly, disclosed herein are combination therapies, and methods of treating diseases and / or conditions with a combination therapy described herein.Description

[0004] Antibody-drug conjugates (ADC) were initially designed to leverage the exquisite specificity of antibodies to deliver targeted potent chemotherapeutic agents with the intention of improving the therapeutic index (the ratio between the toxic dose and the dose at which the drug becomes effective). Unfortunately, the greatest challenge to date for developing ADCs is the therapeutic index is often far narrower than expected. While the FDA approvals of ADCs, such as enfortumab vedotin and tisotumab vedotin, for treatment of certain solid tumors represent milestones in the ongoing development of therapeutic ADCs, there remains a need for improved ADCs and related therapies to help address the long-felt need for additional options to treat different types of cancer and / or to deliver therapeutic payloads to selected cells and / or tissues.SUMMARY

[0005] Some embodiments described herein relate to the use of a combination of Compound (A) and an antibody-drug conjugate for treating a cancer in a subject.Compoundor a pharmaceutically acceptable salt thereof, wherein Ab is an antibody or an antigenbinding fragment thereof, and n represents an average number of vedotin moieties conjugated to a single Ab molecule and is in a range of about 1 to about 10.

[0006] Other embodiments described herein relate to the use of a combination of Compound (A) and an antibody -drug conjugate for treating a cancer in a subject, wherein the antibody-drug conjugate is T-DXd or a pharmaceutically acceptable salt thereof, which is represented by the structure:or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer is selected from HER2-resistant gastric cancer, HER2-refractory gastric cancer, trastuzumab-resistant gastric cancer, trastuzumab-refractory gastric cancer, HER2-resistant esophageal cancer,HER2-refractory esophageal cancer, trastuzumab-resistant esophageal cancer, trastuzumab-refractory esophageal cancer, HER2-resistant gastroesophageal cancer, HER2-refractory gastroesophageal cancer, trastuzumab-resistant gastroesophageal cancer and trastuzumab-refractory gastroesophageal cancer.BRIEF DESCRIPTION OF THE DRAWINGS

[0007] FIGS. 1A and 1C show 2D heatmaps of in vitro synergy between Compound (A) (z.e., azenosertib) and respectively monomethyl auristatin E (MMAE) (FIG. 1A) or tisotumab vedotin (FIG. 1C) in HeLa cervical cancer cells.

[0008] FIGS. IB and ID illustrate protein biomarker changes in response to Compound (A), MMAE (FIG. IB) or tisotumab vedotin (FIG. ID) monotherapy and a combination of Compound (A) and MMAE (FIG. IB) or tisotumab vedotin (FIG. ID) in HeLa cervical cancer cells.

[0009] FIGS. 2A, 2B and 2C present in vivo anti-tumor and tolerability data of mice inoculated with Nectin-4-positive HT-1376 urothelial carcinoma cancer cells, then treated with vehicle (control), azenosertib monotherapy, enfortumab vedotin monotherapy, and azenosertib + enfortumab vedotin combination therapy. FIG.2A is a line plot showing mean tumor volume measurements over the course of 25 days in response to the various aforementioned treatment regimens, with the respective tumor growth inhibition (TGI) percentages measured at day 25 provided. FIG. 2B is a waterfall plot showing tumor volume change percentages at day 25 from the day zero baseline, in response to the various aforementioned treatment regimens. FIG.2C shows mean body weight changes of the mice subject to the treatment regimens of FIG. 2A.

[0010] FIGS. 2D and 2E present in vivo pharmacodynamic data of Nectin-4-positive HT-1376 urothelial carcinoma tumors treated with vehicle (control, V), azenosertib monotherapy (A), enfortumab vedotin monotherapy (EV) and azenosertib + enfortumab vedotin combination therapy (A+EV). FIG. 2D depicts representative immunohistochemistry images showing biomarker staining for phosphorylated CDK1 at tyrosine 15 (pCDKl), gamma-H2AX (yH2AX) and cleaved caspase 3. FIG. 2E is the quantification of positive biomarker staining corresponding to the images and treatment groups shown in FIG 2D.

[0011] FIGS. 3A-3C present in vivo anti-tumor and tolerability data of mice inoculated with tumor fragments derived from a HER2-positive esophageal cancer patient who progressed on T-DXd treatment, then treated with vehicle (control), azenosertibmonotherapy, T-DXd monotherapy and azenosertib + T-DXd combination therapy. FIG.3A is a line plot showing mean tumor volume measurements over the course of 27 days in response to the various aforementioned treatment regimens, with the respective tumor growth inhibition (TGI) percentages measured at day 27. FIG. 3B is a waterfall plot showing tumor volume change percentages at day 27 from the day zero baseline, in response to the various aforementioned treatment regimens. FIG.3C is a line plot depicting mean body weight changes of the mice in response to the various aforementioned treatment regimens over the course of 27 days.DETAILED DESCRIPTIONDefinitions

[0012] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. All patents, applications, published applications and other publications referenced herein arc incorporated by reference in their entirety unless stated otherwise. In the event that there is a plurality of definitions for a term herein, those in this section prevail unless stated otherwise.

[0013] As used herein, an “immunoconjugate” is a conjugate comprising an immunological substance and a drug moiety that are joined together by a linker. Typically, an immunoconjugate is prepared by reacting the immunological substance with a conjugate as defined herein. An “antibody-drug conjugate” (ADC) is one class of immunoconjugate whereby the immunological substance is an antibody or an antigen-binding fragment thereof.

[0014] As used herein, an “antibody” (Ab) is a protein made by the immune system, or a synthetic or engineered variant thereof, that binds to specific sites on cells or tissues. The term “antibody” herein is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multi-specific antibodies (e.g., bispecific antibodies), chimeric antibodies, nanobodies, and antigen-binding fragments so long as they exhibit the desired antigenbinding activity. Monoclonal antibodies are a type of synthetic antibody. In cancer treatment, monoclonal antibodies may kill cancer cells directly, they may block development of tumor blood vessels, and / or they may help the immune system kill cancer cells. Humanized antibodies are antibodies from non-human species whose protein sequences have been modified to increase their similarity to antibody variants producednaturally in humans. For example, a murine monoclonal antibody, 4D5, directed against the extracellular domain of HER2 and a potent inhibitor of growth of human breast cancer cells overexpressing HER2 in vitro and in xenograft models, was humanized by inserting the complementary determining regions of 4D5 into the framework of a consensus human IgGl. The resulting recombinant humanized anti-HER2 mAb was trastuzumab. An antibody typically is composed of two heavy chains and two light chains; and the heavy chain and the light chain typically each contain three complementarity-determining regions (CDRs) immunoglobulin (Ig) hypervariable domains that determine specific antibody binding.

[0015] An “antigen-binding fragment” refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds the antigen to which the intact antibody binds. Examples of antibody fragments include but are not limited to Fv, Fab, Fab’, Fab’-SH, F(ab’)2, diabodies, linear antibodies, single-chain antibody molecules (e.g., scFv), and single-domain antibodies.

[0016] The term “pharmaceutically acceptable salt” refers to a salt of a compound that does not cause significant irritation to an organism to which it is administered and does not abrogate the biological activity and properties of the compound. In some embodiments, the salt is an acid addition salt of the compound. Pharmaceutical salts can be obtained by reacting a compound with inorganic acids such as hydrohalic acid (e.g., hydrochloric acid or hydrobromic acid), a sulfuric acid, a nitric acid and a phosphoric acid (such as 2,3-dihydroxypropyl dihydrogen phosphate). Pharmaceutical salts can also be obtained by reacting a compound with an organic acid such as aliphatic or aromatic carboxylic or sulfonic acids, for example formic, acetic, succinic, lactic, malic, tartaric, citric, ascorbic, nicotinic, methanesulfonic, ethanesulfonic, p-toluensulfonic, trifluoroacetic, benzoic, salicylic, 2-oxopentanedioic or naphthalenesulfonic acid. Pharmaceutical salts can also be obtained by reacting a compound with a base to form a salt such as an ammonium salt, an alkali metal salt, such as a sodium, a potassium or a lithium salt, an alkaline earth metal salt, such as a calcium or a magnesium salt, a salt of a carbonate, a salt of a bicarbonate, a salt of organic bases such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, C1-C7 alkylamine, cyclohexylamine, triethanolamine, ethylenediamine and salts with amino acids such as arginine and lysine. Those skilled in the art understand that when a salt is formed by protonation of a nitrogen-based group (for example, NH2), the nitrogen-based group can beassociated with a positive charge (for example, NH2 can become NH3+) and the positive charge can be balanced by a negatively charged counterion (such as Cl).

[0017] It is understood that, in any compound described herein having one or more chiral centers, if an absolute stereochemistry is not expressly indicated, then each center may independently be of / ^-configuration or ^-configuration or a mixture thereof. Thus, the compounds provided herein may be enantiomerically pure, enantiomerically enriched, racemic mixture, diastereomerically pure, diastereomerically enriched or a stereoisomeric mixture. In addition, it is understood that, in any compound described herein having one or more double bond(s) generating geometrical isomers that can be defined as E or Z, each double bond may independently be E or Z a mixture thereof. Likewise, it is understood that, in any compound described, all tautomeric forms are also intended to be included.

[0018] It is to be understood that where compounds disclosed herein have unfilled valencies, then the valencies arc to be filled with hydrogens or isotopes thereof, e.g., hydrogen-1 (protium) and hydrogen-2 (deuterium).

[0019] It is understood that the compounds described herein can be labeled isotopically. Substitution with isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, such as, for example, increased in vivo half-life or reduced dosage requirements. Each chemical element as represented in a compound structure may include any isotope of said element. For example, in a compound structure a hydrogen atom may be explicitly disclosed or understood to be present in the compound. At any position of the compound that a hydrogen atom may be present, the hydrogen atom can be any isotope of hydrogen, including but not limited to hydrogen- 1 (protium) and hydrogen-2 (deuterium). Thus, reference herein to a compound encompasses all potential isotopic forms unless the context clearly dictates otherwise.

[0020] It is understood that the methods and combinations described herein include crystalline forms (also known as polymorphs, which include the different crystal packing arrangements of the same elemental composition of a compound), amorphous phases, salts, solvates and hydrates. In some embodiments, the compounds described herein exist in solvated forms with pharmaceutically acceptable solvents such as water, ethanol or the like. In other embodiments, the compounds described herein exist in unsolvated form. Solvates contain either stoichiometric or non-stoichiometric amounts of a solvent and may be formed during the process of crystallization with pharmaceutically acceptable solvents such as water, ethanol or the like. Hydrates are formed when the solvent is water oralcoholates are formed when the solvent is alcohol. In addition, the compounds provided herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the compounds and methods provided herein.

[0021] Where a range of values is provided, it is understood that the upper and lower limit, and each intervening value between the upper and lower limit of the range is encompassed within the embodiments.

[0022] Terms and phrases used in this application, and variations thereof, especially in the appended claims, unless otherwise expressly stated, should be construed as open ended as opposed to limiting. As examples of the foregoing, the term ‘including’ should be read to mean “including, without limitation,” “including but not limited to,” or the like; the term “comprising” as used herein is synonymous with “including,” “containing,” or “characterized by,” and is inclusive or open-ended and does not exclude additional, unrccitcd elements or method steps; the term “having” should be interpreted as “having at least;” the term “includes” should be interpreted as “includes but is not limited to;” the term “example” is used to provide exemplary instances of the item in discussion, not an exhaustive or limiting list thereof. In addition, the term “comprising” is to be interpreted synonymously with the phrases “having at least” or “including at least.” When used in the context of a compound, composition or device, the term “comprising” means that the compound, composition or device includes at least the recited features or components but may also include additional features or components.

[0023] With respect to the use of substantially any plural and / or singular terms herein, those having skill in the art can translate from the plural to the singular and / or from the singular to the plural as is appropriate to the context and / or application. The various singular / plural permutations may be expressly set forth herein for sake of clarity. The indefinite article “a” or “an” does not exclude a plurality. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage. Any reference signs in the claims should not be construed as limiting the scope.Compounds and Antibody-Drug Conjugates

[0024] One aspect of this disclosure pertains to the use of a combination of Compound (A) and an antibody -drug conjugate for treating a cancer in a subject (e.g., a patient, a human subject including an adult or a pediatric subject). Compound (A) isazenosertib or ZN-c3), or a pharmaceutically acceptable salt thereof. The antibody -drug conjugate isor a pharmaceutically acceptable salt thereof, whereinis also known as vedotin and this antibody-drug conjugate is referred herein as “the vedotin- based antibody-drug conjugate.” In the vedotin-based antibody-drug conjugate, described herein, Ab is an antibody or an antigen-binding fragment thereof, and n represents an average number of vedotin moieties conjugated to a single Ab molecule and is in a range of about 1 to about 10 e.g., about 1, about 2, about 3, about 4, about 5, about 5, about 6, about 7, about 8, about 9, about 10). In some embodiments, n is in a range of about 2 to about 8. In some embodiments, n is in a range of about 2 to about 6. In some embodiments, n is in a range of about 4 (e.g., 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5).

[0025] Another aspect of this disclosure relates to a method of treating a cancer in a subject comprising administering an effective amount of Compound (A) as defined above and an effective amount of the vedotin-based antibody-drug conjugate as defined above, to the subject.

[0026] Yet another aspect of this disclosure relates to a combination of Compound (A) as defined above and the vedotin-based antibody-drug conjugate as defined above for use in treating a cancer. The use includes administering to a subject suffering from a cancer described herein, a combination of an effective amount of Compound (A) asdefined above and an effective amount of the vedotin-based antibody-drug conjugate as defined above.

[0027] Collectively, all of the foregoing aspects of this disclosure are referred to as “the combination therapy described herein including all aspects and embodiments thereof’ in the ensuing paragraphs.Antibodies

[0028] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, Ab is an antibody or an antigen-binding fragment thereof that specifically binds to a target selected from Nectin-4, tissue factor (TF or also known as platelet tissue factor), zinc transporter LIV-1, sodium-dependent phosphate transporter NaPi2b, integrin beta-6 (ITGB6), B7 homolog 4 (B7-H4) and programmed death- ligand 1 (PD-L1). In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, Ab is an antibody or an antigen-binding fragment thereof that specifically binds to Ncctin-4. In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, Ab is an antibody or an antigen-binding fragment thereof that specifically binds to tissue factor (TF). In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, Ab is an antibody or an antigen-binding fragment thereof that specifically binds to zinc transporter LIV-1. In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, Ab is an antibody or an antigen-binding fragment thereof that specifically binds to sodium-dependent phosphate transporter NaPi2b. In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, Ab is an antibody or an antigenbinding fragment thereof that specifically binds to integrin beta-6 (ITGB6). In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, Ab is an antibody or an antigen-binding fragment thereof that specifically binds to B7 homolog 4 (B7-H4). In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, Ab is an antibody or an antigen-binding fragment thereof that specifically binds to programmed death-ligand 1 (PD-L1).

[0029] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises:(a) a heavy chain CDRH1 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO:1, a heavy chain CDRH2 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO:2 and a heavy chain CDRH3 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO:3; and(b) a light chain CDRL1 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO:4, a light chain CDRL2 comprising or having at least 95% or at least 99% sequence similarity to AAS and a light chain CDRE3 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO:5.

[0030] In some embodiments, a heavy chain CDRH1 comprises or has at least 95% (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO:1, a heavy chain CDRH2 comprises or has at least 95% (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO:2 and / or a heavy chain CDRH3 comprises or has at least 95% (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO:3.

[0031] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises a heavy chain CDRH1 comprises or has at least 99% sequence similarity to SEQ ID NO:1, a heavy chain CDRH2 comprises or has at least 99% sequence similarity to SEQ ID NO:2 and / or a heavy chain CDRH3 comprises or has at least 99% sequence similarity to SEQ ID NO:3. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain CDRH1 comprises the sequence of SEQ ID NO: 1 , a heavy chain CDRH2 comprises the sequence of SEQ ID NO:2 and / or a heavy chain CDRII3 comprises the sequence of SEQ ID NO:3.

[0032] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises a light chain CDRL1 comprises or has at least 95% (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NON, a light chain CDRL2 comprises or has at least 95% (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to AAS and / or a light chain CDRL3 comprises or has at least 95% (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO:5. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a light chain CDRL1 comprises or has at least 99% sequence similarity to SEQ ID NON, a light chain CDRL2 comprises or has at least 99% sequence similarity to AAS and / or a light chain CDRL3 comprises or has at least 99% sequence similarity to SEQ ID NO:5. In some embodiments of the uses andmethods provided herein, the antibody or an antigen-binding fragment comprises a light chain CDRL1 comprises the sequence of SEQ ID NO:4, a light chain CDRL2 comprises the sequence of AAS and / or a light chain CDRL3 comprises the sequence of SEQ ID NO:5.

[0033] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises:(a) a heavy chain CDRH1 comprising SEQ ID NO:1, a heavy chain CDRH2 comprising SEQ ID NO:2 and a heavy chain CDRH3 comprising SEQ ID NO:3; and (b) a light chain CDRL1 comprising SEQ ID NO:4, a light chain CDRL2 comprising AAS and a light chain CDRL3 comprising SEQ ID NO:5.

[0034] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 90%, 95%, or 99% sequence similarity to SEQ ID NO:6, and a light chain comprising having at least 90%, 95%, or 99% sequence similarity to SEQ ID NO:7. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 90% (e.g., at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO:6, and a light chain comprising or having at least 90%, (e.g., at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO:7. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 95% (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO:6, and a light chain comprising or having at least 95%, (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO:7. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 99% sequence similarity to SEQ ID NO:6, and a light chain comprising or having at least 99%, sequence similarity to SEQ ID NO:7. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising the sequence of SEQ ID NO:6, and a light chain comprising the sequence of SEQ ID NO:7.

[0035] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises:(a) a heavy chain CDRH1 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO:8, a heavy chain CDRH2 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO:9 and a heavy chain CDRH3 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO: 10; and(b) a light chain CDRL1 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO: 11, a light chain CDRL2 comprising or having at least 95% or at least 99% sequence similarity to AAS and a light chain CDRE3 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO: 12.

[0036] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises:(a) a heavy chain CDRH1 comprising SEQ ID NO: 8, a heavy chain CDRH2 comprising SEQ ID NO:9 and a heavy chain CDRH3 comprising SEQ ID NO: 10; and (b) a light chain CDRL1 comprising SEQ ID NO: 11, a light chain CDRL2 comprising AAS and a light chain CDRE3 comprising SEQ ID NO: 12.

[0037] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 90%, 95%, or 99% sequence similarity to SEQ ID NO: 13, and a light chain comprising having at least 90%, 95%, or 99% sequence similarity to SEQ ID NO: 14. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 90% (e.g., at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO: 13, and a light chain comprising or having at least 90%, (e.g., at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO: 14. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 95% (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO: 13, and a light chain comprising or having at least 95%, (e.g., at least 95, 96, 97, 98, or 99%) sequence similarity to SEQ ID NO: 14. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 99% sequence similarity to SEQ ID NO: 13, and a light chain comprising or having at least 99%, sequence similarity to SEQ ID NO: 14. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavychain comprising the sequence of SEQ ID NO: 13, and a light chain comprising the sequence of SEQ ID NO: 14.

[0038] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises:(a) a heavy chain CDRH1 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO: 15, a heavy chain CDRH2 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO: 16 and a heavy chain CDRH3 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO: 17; and(b) a light chain CDRL1 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO: 18, a light chain CDRL2 comprising or having at least 95% or at least 99% sequence similarity to WAS and a light chain CDRL3 comprising or having at least 95% or at least 99% sequence similarity to SEQ ID NO: 19.

[0039] In some embodiments, a heavy chain CDRH1 comprises or has at least 95% (e.g., at least 95, 96, 97, 98 or 99%) sequence similarity to SEQ ID NO:15, a heavy chain CDRH2 comprises or has at least 95% (e.g., at least 95, 96, 97, 98 or 99%) sequence similarity to SEQ ID NO:16 and / or a heavy chain CDRH3 comprises or has at least 95% (e.g., at least 95, 96, 97, 98 or 99%) sequence similarity to SEQ ID NO: 17.

[0040] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises a heavy chain CDRH1 comprises or has at least 99% sequence similarity to SEQ ID NO: 15, a heavy chain CDRII2 comprises or has at least 99% sequence similarity to SEQ ID NO: 16 and / or a heavy chain CDRH3 comprises or has at least 99% sequence similarity to SEQ ID NO: 17. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain CDRH1 comprises the sequence of SEQ ID NO: 15, a heavy chain CDRH2 comprises the sequence of SEQ ID NO:16 and / or a heavy chain CDRH3 comprises the sequence of SEQ ID NO:17.

[0041] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises a light chain CDRL1 comprises or has at least 95% (e.g., at least 95, 96, 97, 98 or 99%) sequence similarity to SEQ ID NO: 18, a light chain CDRL2 comprises or has at least 95% (e.g., at least 95, 96, 97, 98 or 99%) sequence similarity to WAS and / or a light chain CDRL3 comprises or has at least 95% (e.g., at least 95, 96, 97, 98 or 99%) sequencesimilarity to SEQ ID NO: 19. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a light chain CDRL1 comprises or has at least 99% sequence similarity to SEQ ID NO: 18, a light chain CDRL2 comprises or has at least 99% sequence similarity to WAS and / or a light chain CDRL3 comprises or has at least 99% sequence similarity to SEQ ID NO: 19. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a light chain CDRL1 comprises the sequence of SEQ ID NO: 18, a light chain CDRE2 comprises the sequence of WAS and / or a light chain CDRL3 comprises the sequence of SEQ ID NO: 19.

[0042] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises:(a) a heavy chain CDRH1 comprising SEQ ID NO: 15, a heavy chain CDRH2 comprising SEQ ID NO: 16 and a heavy chain CDRH3 comprising SEQ ID NO: 17; and (b) a light chain CDRL1 comprising SEQ ID NO: 18, a light chain CDRL2 comprising WAS and a light chain CDRL3 comprising SEQ ID NO: 19.

[0043] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 90%, 95% or 99% sequence similarity to SEQ ID NO:20, and a light chain comprising having at least 90%, 95% or 99% sequence similarity to SEQ ID NO:21. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 90% (e.g., at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%) sequence similarity to SEQ ID NO:20, and a light chain comprising or having at least 90%, (e.g., at least 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%) sequence similarity to SEQ ID NO:21. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 95% (e.g., at least 95, 96, 97, 98 or 99%) sequence similarity to SEQ ID NO:20, and a light chain comprising or having at least 95%, (e.g., at least 95, 96, 97, 98 or 99%) sequence similarity to SEQ ID NO:21. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavy chain comprising or having at least 99% sequence similarity to SEQ ID NO:20, and a light chain comprising or having at least 99%, sequence similarity to SEQ ID NO:21. In some embodiments of the uses and methods provided herein, the antibody or an antigen-binding fragment comprises a heavychain comprising the sequence of SEQ ID NO:20, and a light chain comprising the sequence of SEQ ID NO:21.

[0044] In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, the antibody-drug conjugate is enfortumab vedotin, or a pharmaceutically acceptable salt thereof.

[0045] In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, the antibody-drug conjugate is tisotumab vedotin, or a pharmaceutically acceptable salt thereof.

[0046] In one embodiment, in the combination therapy described herein including all aspects and embodiments thereof, the antibody-drug conjugate is disitamab vedotin, or a pharmaceutically acceptable salt thereof.

[0047] Another aspect of this disclosure pertains to the use of a combination of Compound (A) and an antibody-drug conjugate for treating a cancer in a subject (e.g., a patient, a human subject including an adult or a pediatric subject). Compound (A) is(also known as azenosertib or ZN-c3), or a pharmaceutically acceptable salt thereof. The antibody-drug conjugate isor a pharmaceutically acceptable salt thereof (also known as trastuzumab deruxtecan or T- DXd, whereby the cytotoxic payload DXd or DS-8201 is conjugated or covalently linked to the HER2-targeting antibody trastuzumab). In one embodiment, the antibody-drug conjugate (trasutumab deruxtecan or T-DXd) isor a pharmaceutically acceptable salt thereof, where on average approximately 8 molecules (e.g., 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4 or 8.5) of cytotoxic payload deruxtecan (DXd) are conjugated or covalently linked to the trastuzumab antibody molecule. In some embodiments, the antibody trastuzumab includes a heavy chain amino acid sequence of SEQ ID NO:22 and a light chain sequence of SEQ ID NO:23. In some embodiments, the cancer is selected from HER2-resistant gastric cancer, HER2-refractory gastric cancer, trastuzumab-resistant gastric cancer, trastuzumab-refractory gastric cancer, HER2-resistant esophageal cancer, HER2-refractory esophageal cancer, trastuzumab-resistant esophageal cancer, trastuzumab-refractory esophageal cancer, HER2-resistant gastroesophageal cancer, HER2-refractory gastroesophageal cancer, trastuzumab-resistant gastroesophageal cancer and trastuzumab-refractory gastroesophageal cancer.

[0048] Another aspect of this disclosure relates to a method of treating a cancer in a subject comprising administering an effective amount of Compound (A) as defined above and an effective amount of T-DXd as defined above, to the subject. In some embodiments, the cancer is selected from HER2-resistant gastric cancer, HER2-refractory gastric cancer, trastuzumab-resistant gastric cancer, trastuzumab-refractory gastric cancer, HER2-resistant esophageal cancer, HER2-refractory esophageal cancer, trastuzumabresistant esophageal cancer, trastuzumab-refractory esophageal cancer, HER2-resistant gastroesophageal cancer, HER2 -refractory gastroesophageal cancer, trastuzumab-resistant gastroesophageal cancer and trastuzumab-refractory gastroesophageal cancer.

[0049] Yet another aspect of this disclosure relates to a combination of Compound (A) as defined above and T-DXd as defined above for use in treating a cancer. The use includes administering to a subject suffering from a cancer described herein, a combination of an effective amount of Compound (A) as defined above and an effectiveamount of T-DXd as defined above. The cancer is as defined above and an effective amount of T-DXd as defined above.Cancers

[0050] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the cancer is a solid tumor.

[0051] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, this disclosure relates to the use of a combination of Compound (A) as defined herein, and an antibody-drug conjugate for treating a cancer in a subject. The antibody-drug conjugate can be represented by the following structural formula:or a pharmaceutically acceptable salt thereof, wherein:Ab is an antibody or an antigen-binding fragment thereof that comprises:(a) a heavy chain CDRH1 comprising SEQ ID NO:1, a heavy chain CDRH2 comprising SEQ ID NO:2 and a heavy chain CDRH3 comprising SEQ ID NO:3; and (b) a light chain CDRL1 comprising SEQ ID NO:4, a light chain CDRL2 comprising AAS and a light chain CDRL3 comprising SEQ ID NO:5; andn is in a range of about 1 to about 10 (e.g., about 4).

[0052] In one embodiment, the cancer treated is selected from urogenital cancer, breast cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, lung cancer and cervical cancer. In one embodiment, the urogenital cancer is selected from bladder cancer, cancer of the urinary tract (i.e., ureter, urethra, urothelium), penile cancer, vaginal cancer, renal cancer and testicular cancer. In one embodiment, the cancer treated or the urogenital cancer is bladder cancer, including Nectin-4-positive bladder cancer. In one embodiment, the cancer treated or the urogenital cancer is cancer of the urinary tract (i.e., ureter, urethra, urothelium), including Nectin-4-positive cancer of the urinary tract. In one embodiment, the cancer treated or the urogenital cancer is urothelial carcinoma, including Nectin-4-positive urothelial carcinoma.

[0053] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, this disclosure relates to the use of acombination of Compound (A) as defined herein, and an antibody-drug conjugate for treating a cancer in a subject. The antibody-drug conjugate can be represented by the following structural formula:or a pharmaceutically acceptable salt thereof, wherein:Ab is an antibody or an antigen-binding fragment thereof that comprises:(a) a heavy chain CDRH1 comprising SEQ ID NO:8, a heavy chain CDRH2 comprising SEQ ID NO:9 and a heavy chain CDRH3 comprising SEQ ID NO: 10; and (b) a light chain CDRL1 comprising SEQ ID NO: 11, a light chain CDRL2 comprising AAS and a light chain CDRL3 comprising SEQ ID NO: 12; andn is in a range of about 1 to about 10 (e.g., about 4).

[0054] In one embodiment, the cancer treated is selected from cervical cancer, breast cancer, pancreatic cancer, head and neck cancer and lung cancer. In one embodiment, the cancer treated or the urogenital cancer is cervical cancer, including tissue factor (TF)- positive cervical cancer.Prior Treatment Regimens

[0055] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the subject has received at least one (e.g., one, two or three) prior treatment regimen. Such prior treatment regimen may be given at locally advanced setting, metastatic setting, neoadjuvant setting or adjuvant setting. In some embodiments, such prior treatment regimen that acts as first- line standard therapies comprises systemic therapy, oral therapy, radiation therapy, surgery, or a combination thereof. Non-limiting examples of such systemic therapy include immunotherapy, targeted therapy, chemotherapy, chemoradiotherapy (also known as radiochemotherapy), or a combination thereof. Similarly, non-limiting examples of such oral therapy include immunotherapy (e.g. , immune checkpoint inhibitors), targeted therapy, chemotherapy (e.g. , platinum-based or platinum containing chemotherapy), chemoradiotherapy, or a combination thereof.

[0056] Non-limiting examples of immune checkpoint inhibitors include PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors and LAG-3 inhibitors (including pharmaceutically acceptable salts of any of the foregoing). In some embodiments, an immune checkpoint inhibitor may be bi- or multi -specific and may target at least two proteins, including, but not limited to, two proteins selected from PD-1, PD-L1, CTLA-4 and LAG-3. Non-limiting examples of PD-1 inhibitors include retifanlimab, pucotenlimab, cadonilimab, serplulimab, nivolumab (relatimab), zimberelimab, penpulimab, dostarlimab (dostarlimab-gxly), prolgolimab, tislelizumab, camrelizumab, sintilimab, toripalimab, cemiplimab (cemiplimab-RWLC), pembrolizumab, nivolumab, balstilimab, finotonlimab, iparomlimab, ivonescimab, tuvonralimab / iparomlimab, cetrelimab, favezelimab / pembrolizumab, genolimzumab, nofazinlimab, pembrolizumab / hyaluronidase, pembrolizumab / quavonlimab, pembrolizumab / vibostolimab, pembrolizumab / quavonlimab, rilvegostomig, sasanlimab, spaitalizumab, tcbotclimab and volrustomig (including pharmaceutically acceptable salts of any of the foregoing).

[0057] Non-limiting examples of PD-L1 inhibitors include socazolimab, adebrelimab, sugemalimab, envafolimab, durvalumab, avelumab, atezolizumab, benmelstobart, tagitanlimab, bintrafusp alfa, erfonrilimab and retlirafusp alfa (including pharmaceutically acceptable salts of any of the foregoing).

[0058] Non-limiting examples of CTLA-4 inhibitors include tremelimumab, cadonilimab, ipilimumab, tuvonralimab / paromlimab, erfonrilimab, gotistobart, pembrolizumab / quavonlimab, pembrolizumab / quavonlimab, quavonlimab and volrustomig (including pharmaceutically acceptable salts of any of the foregoing).

[0059] Non-limiting examples of LAG-3 inhibitors include nivolumab / relatlimab, favezelimab / pembrolizumab, fianlimab, relatlimab, tebotelimab, eftilagimod alpha and favezelimab (including pharmaceutically acceptable salts of any of the foregoing).

[0060] In some embodiments, in the combination therapy described herein including all aspects and embodiments thereof, the subject has received at least prior treatment regimen selected from a PD-1 inhibitor, a PD-L1 inhibitor and a platinum-containing chemotherapy. Non-limiting examples of platinum-based chemotherapy include cisplatin, carboplatin and oxaliplatin (including pharmaceutically acceptable salts of any of the foregoing).General Dosing Regimens and Effects of Combination Therapy

[0061] When the treatment is a combination of compounds, the order of administration or use of the combination of Compound (A) and the vedotin-based antibodydrug conjugate described herein can vary. In some embodiments, Compound (A) and the vedotin-based antibody-drug conjugate are administered or used sequentially. In some embodiments, Compound (A) can be administered prior to the vedotin-based antibody-drug conjugate. In other embodiments, Compound (A) can be administered subsequent to or after the vedotin-based antibody-drug conjugate. In still other embodiments, Compound (A) can be administered concomitantly or concurrently with the vedotin-based antibodydrug conjugate.

[0062] In some embodiments, Compound (A) is provided on an intermittent dosing schedule. In some embodiments, Compound (A) the intermittent dosing schedule comprises 5 days on, 2 days off; 4 days on, 3 days off; 3 days on, 4 days off; or 6 days on, 1 day off on in one or more dosing weeks.

[0063] In some embodiments, the use of a combination of Compound (A) and the vedotin-based antibody-drug conjugate for treating a cancer or methods of treatment with a combination of Compound (A) and the vedotin-based antibody-drag conjugate provided herein can decrease the number and / or severity of side effects that can be attributed to monotherapy of the vedotin-based antibody-drag conjugate.

[0064] Using a combination of Compound (A) and the vedotin-based antibodydrug conjugate as described herein can result in additive, synergistic or strongly synergistic effect. A combination of compounds described herein can result in an effect that is not antagonistic.

[0065] As used herein, the term “antagonistic” means that the activity of the combination of compounds is less compared to the sum of the activities of the compounds in combination when the activity of each compound is determined individually (i.e., as a single compound). As used herein, the term “synergistic effect” means that the activity of the combination of compounds is greater than the sum of the individual activities of the compounds in the combination when the activity of each compound is determined individually. As used herein, the term “additive effect” means that the activity of the combination of compounds is about equal to the sum of the individual activities of the compounds in the combination when the activity of each compound is determined individually.

[0066] A potential advantage of utilizing a combination as described herein may be a reduction in the required amount(s) of the compound(s) that is effective in treating a disease condition disclosed herein compared to when Compound (B) or the vedotin-based antibody-drug conjugate is administered as a monotherapy and / or wherein Compound (A) is administered as a monotherapy. For example, the amount of Compound (A) and the vedotin-based antibody-drug conjugate used in a combination described herein can be less compared to the amount of Compound (A) and the vedotin-based antibody-drug conjugate needed to achieve the same reduction in a disease marker (for example, tumor size) when each is administered as a monotherapy. Another potential advantage of utilizing a combination as described herein is that the use of two or more compounds having different mechanisms of action can create a higher barrier to the development of resistance compared to when Compound (A) and the vedotin-based antibody-drug conjugate are each administered as monotherapy. Additional advantages of utilizing a combination as described herein may include little to no cross resistance between the compounds of a combination described herein; different routes for elimination of the compounds of a combination described herein; and / or little to no overlapping toxicities between the compounds of a combination described herein.Pharmaceutical Compositions

[0067] Compound (A) can be provided in a pharmaceutical composition. Likewise, the vedotin-based antibody-drug conjugate can be provided in a pharmaceutical composition. In some embodiments, Compound (A) and the vedotin-based antibody-drug conjugate can be provided in the same pharmaceutical composition. In other embodiments, Compound (A) and the vedotin-based antibody-drug conjugate can be provided in separate pharmaceutical compositions.

[0068] The term “pharmaceutical composition” refers to a mixture of one or more compounds and / or salts disclosed herein with other chemical components, such as diluents, carriers and / or excipients. The pharmaceutical composition facilitates administration of the compound to an organism. Pharmaceutical compositions can also be obtained by reacting compounds with inorganic or organic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicylic acid. Pharmaceutical compositions will generally be tailored to the specific intended route of administration.

[0069] The pharmaceutical compositions described herein can be administered to a human patient per se, or in pharmaceutical compositions where they are mixed withother active ingredients, as in combination therapy, or carriers, diluents, excipients or combinations thereof. Proper formulation is dependent upon the route of administration chosen. Techniques for formulation and administration of the compounds described herein are known to those skilled in the art.

[0070] The pharmaceutical compositions disclosed herein may be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tableting processes. Additionally, the active ingredients are contained in an amount effective to achieve its intended purpose. Many of the compounds used in the pharmaceutical combinations disclosed herein may be provided as salts with pharmaceutically compatible counterions.

[0071] Multiple techniques of administering a compound, salt and / or composition exist in the art including, but not limited to, oral, rectal, pulmonary, topical, aerosol, injection, infusion and parenteral delivery, including intramuscular, subcutaneous, intravenous, intramedullary injections, intrathecal, direct intraventricular, intraperitoneal, intranasal and intraocular injections. In one embodiment, Compound (A) is administered orally. In one embodiment, the vedotin-based antibody-drug conjugate is administered intravenously.Miscellaneous

[0072] In some cases, following cancer treatment, a subject can relapse or have reoccurrence of the cancer. As used herein, the terms “relapse” and “reoccurrence” are used in their normal sense as understood by those skilled in the art. Thus, in some embodiments, the cancer can be a recurrent cancer. Furthermore, in some embodiments, in a combination therapy described herein including all aspects and embodiments thereof, the cancer treated is unresectable. In some embodiments, in a combination therapy described herein including all aspects and embodiments thereof, the cancer treated is locally advanced or metastatic.

[0073] As used herein, the terms “treat,” “treating,” “treatment,” “therapeutic,” and “therapy” do not necessarily mean total cure or abolition of the disease or condition. Any alleviation of any undesired signs or symptoms of the disease or condition, to any extent can be considered treatment and / or therapy. Furthermore, treatment may include acts that may worsen the subject’s overall feeling of well-being or appearance.

[0074] The term “effective amount” is used to indicate an amount of an active compound, or pharmaceutical agent, that elicits the biological or medicinal responseindicated. For example, an effective amount of compound, salt or composition can be the amount needed to prevent, alleviate or ameliorate symptoms of the disease or condition, or prolong the survival of the subject being treated. This response may occur in a tissue, system, animal or human and includes alleviation of the signs or symptoms of the disease or condition being treated. Determination of an effective amount is well within the capability of those skilled in the art, in view of the disclosure provided herein. The effective amount of the compounds disclosed herein required as a dose will depend on the route of administration, the type of animal, including human, being treated and the physical characteristics of the specific animal under consideration. The dose can be tailored to achieve a desired effect, but will depend on such factors as weight, diet, concurrent medication and other factors which those skilled in the medical arts will recognize.

[0075] For example, an effective amount of a compound, or radiation, is the amount that results in: (a) the reduction, alleviation or disappearance of one or more symptoms caused by the cancer, (b) the reduction of tumor size, (c) the elimination of the tumor, and / or (d) long-term disease stabilization (growth arrest) of the tumor.

[0076] The amount of compound, salt and / or composition required for use in treatment will vary not only with the particular compound or salt selected but also with the route of administration, the nature and / or symptoms of the disease or condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. In cases of administration of a pharmaceutically acceptable salt, dosages may be calculated as the free base. As will be understood by those of skill in the art, in certain situations it may be necessary to administer the compounds disclosed herein in amounts that exceed, or even far exceed, the dosage ranges described herein in order to effectively and / or aggressively treat particularly aggressive diseases or conditions.

[0077] It should be noted that the attending physician would know how to and when to terminate, interrupt or adjust administration due to toxicity or organ dysfunctions. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response was not adequate (precluding toxicity). The magnitude of an administrated dose in the management of the disorder of interest will vary with the severity of the disease or condition to be treated and to the route of administration. The severity of the disease or condition may, for example, be evaluated, in part, by standard prognostic evaluation methods. Further, the dose and perhaps dose frequency, will also vary accordingto the age, body weight and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.

[0078] Compounds, salts and compositions disclosed herein can be evaluated for efficacy and toxicity using known methods. For example, the toxicology of a particular compound, or of a subset of the compounds, sharing certain chemical moieties, may be established by determining in vitro toxicity towards a cell line, such as a mammalian, and preferably human, cell line. The results of such studies are often predictive of toxicity in animals, such as mammals, or more specifically, humans. Alternatively, the toxicity of particular compounds in an animal model, such as mice, rats, rabbits, dogs or monkeys, may be determined using known methods. The efficacy of a particular compound may be established using several recognized methods, such as in vitro methods, animal models, or human clinical trials. When selecting a model to determine efficacy, the skilled artisan can be guided by the state of the art to choose an appropriate model, dose, route of administration and / or regime.EXAMPLES

[0079] Additional embodiments are disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the claims. In these examples and in the accompanying figures, Compound (A) is azenosertib and is also referred to as the same.Example 1 : Azenosertib in combination with MMAE or Tisotumab vedotin exhibited in vitro synergy and increased in vitro apoptosis

[0080] HeLa cervical cancer cells (tissue factor-positive or TF+) were cultured in EMEM medium (ATCC) supplemented with 10% FBS (Gibco) and 1% penicillinstreptomycin (Gibco). Cells were maintained at 37°C in an atmosphere containing 5% CO2.

[0081] HeLa cells were seeded at optimal density in 96-well plates for evaluation of combination treatments on cell viability. Treatments were performed the following day with azenosertib (0-1000 nM) or MMAE (0-1 nM) or tisotumab vedotin (0-1000 nM) as monotherapies or in a matrix combination format. Cell viability was then measured 2 or 3 days following treatment by CellTiter-Glo 2.0 assay. Synergies between azenosertib and MMAE or between azenosertib and tisotumab vedotin were then calculated using ZIP synergy score from SynergyEinder tool (http: / / www.synergyfinderplus.org / ).

[0082] HeLa cells were seeded at optimal density in 6 cm dishes for the evaluation of combination treatments on biomarker changes. Treatment doses were determined from combination CellTiter-Glo assays and treatments were performed the following day with azenosertib (300 nM) and MMAE (0.5 nM) or azenosertib (250 nM) and tisotumab vedotin (250 nM) as monotherapies or as combination therapy. After indicated time of drug exposure, cells were harvested, rinsed with PBS, lysed and sonicated in cold RIPA buffer (Sigma) containing protease and phosphatase inhibitors for 15 minutes, and centrifuged at 4°C, 15,000 rpm for 15 minutes. Protein concentrations were measured using the Pierce BCA Protein Assay kit (Thermo Fisher). JESS plates were loaded and run according to the manufacturer’s instructions. Primary antibodies anti-Cyclin El was purchased from Abeam, anti-GAPDH was purchased from Invitrogen, anti-phospho-CDK1, anti-cleaved caspase 3, anti-Weel, and anti-CDKl were purchased from Cell Signaling Technology.

[0083] As shown in FIGS.1A and 1C, azenosertib in combination with MMAE or tisotumab vedotin both showed synergy in HeLa cells, i.e., demonstrated a clear synergy (ZIP synergy score > 10) at depicted dose ranges, with increased apoptosis as indicated by cleaved caspase 3 (see FIGS. IB and ID).Example 2: Azenosertib in combination with enfortumab vedotin enhanced tumor growth inhibition in vivo

[0084] The HT-1376 urothelial carcinoma (Nectin-4 positive, 77J53-rnulaled) tumor cell line was maintained in vitro as monolayer culture in EMEM supplemented with 10% heat inactivated fetal bovine serum at 37°C in an atmosphere of 5% CO2 in air. The tumor cells were routinely sub-cultured before confluence by trypsin-EDTA treatment, not to exceed 4-5 passages. The cells growing in an exponential growth phase were harvested and counted for tumor inoculation.

[0085] NOD SCID female mice (6 to 8 weeks) were purchased from Beijing Anikeeper Biotech Co., Ltd. Each mouse was inoculated subcutaneously on the right flank with single cell suspension of tumor cells (1 x 107) in 100 pL EMEM and Matrigel mixture (1:1 ratio) without serum for the tumor development. The treatments were administered to the tumor-bearing mice (n = 8 in each group) according to the study design shown in Table 1.Table 1. HT-1376 Nectin-4+Urothelial Cancer Study design

[0086] Body weights of animals were measured twice weekly throughout the study. Body weight change, expressed in %, was calculated using the following formula: BW change (%) = ((BW Day X “ BW D y o) / BW Day o) x 100, where BW Day X is BW on a given day, and BW Day o is BW on Day 0 (initiation of treatment).

[0087] The measurement of tumor size was conducted twice a week with a caliper and the tumor volume (mm3) was estimated using the formula: TV = a x b2 / 2 throughout the study, where “a” and “b” were long and short diameters of a tumor, respectively. The TVs were used for calculation of the tumor growth inhibition (TGI, an indicator of antitumor effectiveness) value using the formula: (1 - (Ta - To) I (Ca - Co)) x 100%. Ta and Ca were the mean tumor volumes of the treated and control animals, and To and Co were the mean tumor volumes of the treated and control animals at the start of the experiment.

[0088] The combination therapy of azenosertib and enfortumab vedotin enhanced tumor growth inhibition (see FIG. 2A) and induced tumor regressions that were not observed in their respective monotherapies, when the tumor volumes were measured at day 25 (see FIG. 2B). In FIG. 2C, the body weight change measurements are shown and indicate minimal body weight changes throughout the study in this exemplary combination therapy. Typically, a body weight loss of greater than 15% indicates that the treatment regimen is not well-tolerated. As shown in FIG. 2C, the combination therapy of azenosertib and enfortumab vedotin was well-tolerated.

[0089] HT-1376 tumors were collected from mice at end-of-study, 4 hours postfinal dose. Tumors were formalin-fixed and paraffin-embedded for immunohistochemistry analysis of downstream pharmacodynamic biomarkers (FIG 2D). Biomarker staining wasquantified using H-score (FIG 2E). Decreased pCDKl, the proximal biomarker for WEE1 activity, was observed in tumors treated with azenosertib or azenosertib + enfortumab vedotin, confirming that azenosertib exerted on-target inhibition of WEE1 and its activity. Increased \H2AX, a marker of DNA damage and replication stress, was observed in all drug-treated groups and was significantly higher in tumors treated with the combination of azenosertib + enfortumab vedotin. Likewise, cleaved caspase 3, a marker of apoptosis, was also significantly elevated in tumors treated with the combination. These data demonstrate that the anti-tumor activity of combining azenosertib + enfortumab vedotin is driven by increased DNA damage and apoptosis.Example 3 In vivo anti-tumor efficacy of azenosertib in combination with trastuzumab deruxtecan (T-DXd) in a patient-derived xenograft (PDX) esophageal cancer model that progressed on T-DXd treatment

[0090] A HER2+ esophageal cancer model derived from a patient who progressed after treatment with T-DXd was used in this study. Tumor fragments were inoculated subcutaneously and once the mean tumor volume reached 235 mm3, mice were randomized and treated with vehicle, azenosertib, T-DXd or the combination of azenosertib and T-DXd. Table 2 summarizes the groups and treatments. In summary, in all applicable groups, azenosertib was administered on an intermittent dosing schedule of 5 days on (QD or once a day) followed by 2 days off while T-DXd was administered once every three weeks (Q3W).Table 2. HER2+ Esophageal Cancer PDX Study Design

[0091] Tumor volume (TV) measurements and animal body weight (BW) were monitored twice weekly through the study. As shown in FIGS. 3A and 3B, the tumors were, as expected, relatively unresponsive to T-DXd monotherapy, only resulting in 27%tumor growth inhibition (TGI) at day 27. Indeed, the tumors were less responsive to T-DXd monotherapy than azenosertib monotherapy, which resulted in 56% TGI. Surprisingly, the combination of azenosertib and T-DXd overcame the resistance to T-DXd, resulting in a significantly higher TGI of 99%. Notably, as can be seen in the waterfall plot in FIG. 3B, at least one of the mice achieved regression at day 27, where the tumor volume percentage change fell below the zero baseline. Furthermore, FIG. 3C shows that the combination of azenosertib and T-DXd was well-tolerated in that all of the mice treated with this combination experienced a body weight loss <1 %. Taken together, these data demonstrate the potential of the azenosertib and T-DXd combination in at least patient populations who were previously insensitive (i.e., inherently resistant) or developed resistance to T-DXd monotherapy and / or any other anti-HER2-based regimen or trastuzumab-based regimen.

[0092] Furthermore, although the foregoing has been described in some detail by way of illustrations and examples for purposes of clarity and understanding, it will be understood by those of skill in the art that numerous and various modifications can be made without departing from the spirit of the present disclosure. Therefore, it should be clearly understood that the forms disclosed herein are illustrative only and are not intended to limit the scope of the present disclosure, but rather to also cover all modification and alternatives coming with the true scope and spirit of the present disclosure.SEQUENCE LISTINGSSEQ ID NO: 1 - Enfortumab CDRII1GFTFSSYN SEQ ID NO: 2 - Enfortumab CDRH2ISSSSSTI SEQ ID NO: 3 Enfortumab CDRH3ARAYYYGMDV SEQ ID NO:4 - Enfortumab CDRL1QGISGW AAS - Enfortumab CDRL2SEQ ID NO: 5 - Enfortumab CDRL3QQANSFPPT SEQ ID NO:6 - Enfortumab heavy chain EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYNMNWVRQAPGKGLEWVSYISSS SSTIYYADSVKGRFTISRDNAKNSLSLQMNSLRDEDTAVYYCARAYYYGMDVW GQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA EPAPIEKTISKAKGQPREPQVYTEPPSREEMTKNQVSETCEVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK SEQ ID NO:7 - Enfortumab light chain DIQMTQSPSSVSASVGDRVTITCRASQGISGWLAWYQQKPGKAPKFLIYAASTLQ SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPPTFGGGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 8 - Tisotumab CDRH1GFTFSNYA SEQ ID NO:9 Tisotumab CDRH2ISGSGDYT SEQ ID NO: 10 - Tisotumab CDRH3ARSPWGYYLDS SEQ ID NO:11 - Tisotumab CDRL1QGISSR AAS - Tisotumab CDRL2SEQ ID NO: 12 - Tisotumab CDRL3QQYNSYPYT SEQ ID NO: 13 - Tisotumab heavy chain EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GDYTYYTDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPWGYYLDS WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY1CNVNHKPSNTKVDKRVE PKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPGK SEQ ID NO: 14 - Tisotumab light chain DIQMTQSPPSLSASAGDRVTITCRASQGISSRLAWYQQKPEKAPKSLIYAASSLQS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPYTFGQGTKLEIKRTVAA PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 15 - Disitamab CDRH1GYTFTDYY SEQ ID NO: 16 - Disitamab CDRH2VNPDHGDS SEQ ID NO: 17 - Disitamab CDRH3ARNYLFDH SEQ ID NO: 18 - Disitamab CDRL1QDVGTA WAS - Disitamab CDRL2SEQ ID NO: 19 - Disitamab CDRL3HQFATYT SEQ ID NO:20 - Disitamab heavy chain EVQLVQSGAEVKKPGATVK1SCKVSGYTFTDYY1HWVQQAPGKGLEWMGRVNP DHGDSYYNQKFKDKATITADKSTDTAYMELSSLRSEDTAVYFCARNYLFDHWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPK SCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKSEQ ID N0:21 - Disitamab light chain DIQMTQSPSSVSASVGDRVTITCKASQDVGTAVAWYQQKPGKAPKLLIYWASIR IITGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCIIQFATYTFGGGTKVEIKRTVAA PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO:22 - Trastuzumab heavy chain EVQLVESGGGLVQPGGSLRESCAASGFNIKDTYIHWVRQAPGKGEEWVARIYPT NGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYA MDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKSEQ ID NO:23 Trastuzumab light chain DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLY S GVPSRFS GSRS GTDFTLTIS SLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Claims

WHAT IS CUAIMED IS:

1. Use of a combination of Compound (A) and an antibody-drug conjugate for treating a cancer in a subject, wherein:acceptable salt thereof;the antibody-drug conjugate isor a pharmaceutically acceptable salt thereof, wherein:Ab is an antibody or an antigen-binding fragment thereof; andn represents an average number of vedotin moieties conjugated to a single Ab molecule and is in a range of about 1 to about 10.

2. The use of claim 1, wherein Ab is an antibody or an antigen-binding fragment thereof that specifically binds to a target selected from Nectin-4, tissue factor (TF or also known as platelet tissue factor), zinc transporter LIV-1, sodium-dependent phosphate transporter NaPi2b, integrin beta-6 (ITGB6), B7 homolog 4 (B7-H4) and programmed death-ligand 1 (PD-L1).

3. The use of claim 1 or 2, wherein Ab is an antibody or an antigen-binding fragment thereof that specifically binds to Nectin-4.

4. The use of claim 1 or 2, wherein Ab is an antibody or an antigen-binding fragment thereof that specifically binds to tissue factor (TF).

5. Use of a combination of Compound (A) and an antibody-dmg conjugate for treating a cancer in a subject, wherein:Compound (acceptable salt thereof;the antibody-drug conjugate isor a pharmaceutically acceptable salt thereof, wherein:Ab is an antibody or an antigen-binding fragment thereof that specifically binds to Nectin-4; andn represents an average number of vedotin moieties conjugated to a single Ab molecule and is in a range of about 1 to about 10.

6. Use of a combination of Compound (A) and an antibody-drug conjugate for treating a cancer in a subject, wherein:acceptable salt thereof;the antibody-drug conjugate isor a pharmaceutically acceptable salt thereof, wherein:Ab is an antibody or an antigen-binding fragment thereof that specifically binds to tissue factor (TF or also known as platelet tissue factor); andn represents an average number of vedotin moieties conjugated to a single Ab molecule and is in a range of about 1 to about 10.

7. The use of any one of claims 1 to 3 or 5, wherein the antibody or an antigenbinding fragment thereof comprises;(a) a heavy chain CDRH1 comprising SEQ ID NO:1, a heavy chain CDRH2 comprising SEQ ID NO: 2 and a heavy chain CDRH3 comprising SEQ ID NO:3; and (b) a light chain CDRL1 comprising SEQ ID NO:4, a light chain CDRL2 comprising AAS and a light chain CDRL3 comprising SEQ ID NO:5.

8. The use of any one of claims 1 to 3, 5, and 7, wherein the antibody or an antigen-binding fragment thereof comprises a heavy chain comprising SEQ ID NO: 6, and a light chain comprising SEQ ID NO:7.

9. The use of any one of claims 1, 2, 4 and 6, wherein the antibody or an antigen-binding fragment thereof comprises:(a) a heavy chain CDRH1 comprising SEQ ID NO:8, a heavy chain CDRH2 comprising SEQ ID NO:9 and a heavy chain CDRH3 comprising SEQ ID NOTO; and (b) a light chain CDRL1 comprising SEQ ID NO: 11, a light chain CDRL2 comprising AAS and a light chain CDRL3 comprising SEQ ID NO: 12.

10. The use of any one of claims 1, 2, 4, 6, and 9, wherein the antibody or an antigen-binding fragment thereof comprises a heavy chain comprising SEQ ID NO: 13, and a light chain comprising SEQ ID NO: 14.

11. The use of any one of claims 1 to 10, wherein n is in a range of about 4.

12. The use of any one of claims 1 to 3, 5, 7, 8 or 11, wherein the antibodydrug conjugate is enfortumab vedotin, or a pharmaceutically acceptable salt thereof.

13. The use of any one of claims 1, 2, 4, 6 or 9 to 11, wherein the antibodydrug conjugate is tisotumab vedotin, or a pharmaceutically acceptable salt thereof.

14. The use of any one of claims 1 to 13, wherein the cancer is a solid tumor.

15. The use of any one of claims 1 to 3, 5, 7, 8, 11, 12 or 14, wherein the cancer is selected from a urogenital cancer, breast cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, lung cancer and cervical cancer.

16. The use of claim 15, wherein the urogenital cancer is selected from bladder cancer, cancer of the urinary tract (i.e., ureter, urethra, urothelium), penile cancer, vaginal cancer, renal cancer and testicular cancer.

17. The use of any one of claims 1 to 3, 5, 7, 8, 11, 12 or 14 to 16, wherein the cancer is selected from bladder cancer and cancer of the urinary tract.

18. The use of any one of claims 1, 2, 4, 6, 9 to 11, 13 or 14, wherein the cancer is selected from cervical cancer, breast cancer, pancreatic cancer, head and neck cancer and lung cancer.

19. The use of any one of claim 1, 2, 4, 6, 9 to 12 or 16, wherein the cancer is cervical cancer.

20. The use of any one of claims 1 to 19, wherein the subject has received at least one prior treatment regimen.

21. The use of claim 20, wherein the at least one prior treatment regimen comprises systemic therapy, oral therapy, radiation therapy, surgery or a combination thereof.

22. The use of claim 21, wherein the systemic therapy and the oral therapy each comprise immunotherapy, targeted therapy, chemotherapy, chemoradiotherapy or a combination thereof.

23. The use of any one of claims 1 to 3, 5, 7, 8, 11, 12, 14 to 16 or 20 to 22, wherein the subject has received at least prior treatment regimen selected from a PD-1 inhibitor, a PD-L1 inhibitor and a platinum-containing chemotherapy.

24. The use of any one of claims 20 to 23, wherein the subject has received at least one prior treatment regimen at locally advanced setting, metastatic setting, neoadjuvant setting or adjuvant setting.

25. U se of a combination of compounds in the manufacture of a medicament for treating a cancer, wherein the combination includes an effective amount of Compound (A), or a pharmaceutically acceptable salt thereof, and an effective amount of an antibodydrug conjugate, or a pharmaceutically acceptable salt thereof, wherein:Compoundpharmaceutically acceptable salt thereof;the antibody-drug conjugateor a pharmaceutically acceptable salt thereof ; andthe cancer is selected from HER2-resistant gastric cancer, HER2-refractory gastric cancer, trastuzumab-resistant gastric cancer, trastuzumab-refractory gastric cancer, HER2- resistant esophageal cancer, HER2-refractory esophageal cancer, trastuzumab-resistant esophageal cancer, trastuzumab-refractory esophageal cancer, HER2-resistant gastroesophageal cancer, HER2-refractory gastroesophageal cancer, trastuzumab-resistant gastroesophageal cancer and trastuzumab-refractory gastroesophageal cancer.

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