Wee1 inhibitor combination therapy

Abstract

Disclosed herein is use of a combination of Compound (A) and Compound (B) as defined herein, for treating a cancer, such as a colorectal, a pancreatic and / or a non-small cell lung cancer.

Description

ZENO.186WO PATENTWEE1 INHIBITOR COMBINATION THERAPYINCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS

[0001] Any and all applications for which a foreign or domestic priority' claim is identified m 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 / 733,574, filed December 13, 2024, which is incorporated by reference in its entirety' including any drawings.Field

[0002] 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

[0003] Oncogenic mutations in RAS (KRAS, HRAS, and NRAS) proto-oncogenes drive up to 30% of human cancers, accounting for more than 200,000 new cancer cases in the United States each year, most notably of non-small cell lung cancer (NSCLC), colorectal cancers, and pancreatic ductal adenocarcinoma (PDAC). Most oncogenic RAS mutations are gain-of-function missense alterations at hotspot codons including, but not limited to, codons 12, 13, and 61, that result in an impairment of GTP hydrolysis and / or acceleration of GDP-to-GTP nucleotide exchange by' these small GTPases, such that the normally tightly regulated cellular equilibrium of a RAS protein shifts predominantly toward the active, GTP-bound (RAS(ON)) state. This shift drives increased oncogenic flux via activation of downstream effectors and signaling pathways linked to cell proliferation and survival. Until the recent development of direct inhibitors of KRAS G12C, RAS was largely considered undruggable.

[0004] KRAS G12C mutant-selective inhibitors introduce an allele-specific covalent modification of the cysteine residue of the KRASG12C protein in the GDP-bound inactive [KRAS G12C(OFF)] state. However, KRAS G12C inhibitors only cover a small fraction of all oncogenic RAS mutations, including the most prevalent codon 12 mutationsother than G12C, leaving a significant unmet medical need for inhibitors targeting most RAS alterations in cancer. Moreover, while KRAS G12C-specific inhibitors, such as sotorasib and adagrasib, are approved and / or recommended for treatment of metastatic NSCLC and CRC, and while responses can be robust, some patients are inherently resistant and most responders acquire resistance. In the pivotal studies that resulted in the approval of the KRAS G12C inhibitors sotorasib and adagrasib, monotherapy resulted in significant responses in non-small lung cell cancer and colorectal cancer. Whilst both compounds demonstrated robust initial clinical responses, patients eventually progressed after -6.5 months on average.SUMMARY

[0005] Some embodiments described herein relate to the use of a combination of Compound (A) and Compound (B) for treating a cancer. Compound (A) isor a pharmaceutically acceptable salt thereof., or a pharmaceutically acceptable salt thereof.BRIEF DESCRIPTION OF THE DRAWINGS

[0006] FIGS. 1A and IB show 2D in vitro dose response curves of Compound (A) (FIG. 1A) or Compound (B) (FIG. IB), FIG. 1C shows a heatmap of 2D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. ID shows a heatmap of 2D in vitro Loewe synergy between Compound(A) and Compound (B). FIGS. 1A-1D were performed in the NCI-H1792 NSCLC cell line harboring the KRAS G12C mutation.

[0007] FIGS. 2A and 2B show 3D in vitro dose response curves of Compound (A) (FIG. 2A) or Compound (B) (FIG. 2B). FIG. 2C shows a heatmap of 3D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 2D shows a heatmap of 2D in vitro Loewe synergy between Compound (A) and Compound (B). FIGS. 2A-2D were performed in the NCI-H1792 NSCLC cell line harboring the KRAS Gl 2C mutation.

[0008] FIGS. 3A and 3B show 2D in vitro dose response curves of Compound (A) (FIG. 3A) or Compound (B) (FIG. 3B). FIG. 3C show's a heatmap of 2D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 3D shows a heatmap of 2D in vitro Loewe synergy between Compound (A) and Compound (B), FIGS. 3A-3D were performed in the SW403 CRC cell line harboring the KRAS G12V mutation.

[0009] FIGS. 4A and 4B show 3 D in vitro dose response curves of Compound (A) (FIG. 4A) or Compound (B) (FIG. 4B). FIG. 4C show's a heatmap of 3D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 4D shows a heatmap of 2D in vitro Loewe synergy between Compound (A) and Compound (B). FIGS. 4A-4D were performed in the SW403 CRC cell line harboring the KRAS G12V mutation.

[0010] FIGS. 5A and 5B show 2D in vitro dose response curves of Compound (A) (FIG. 5A) or Compound (B) (FIG. 5B). FIG. 5C show's a heatmap of 2D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 5D shows a heatmap of 2D in vitro Loewe synergy between Compound (A) and Compound (B). FIGS. 5A-5D were performed in the NCI-H441 NSCLC cell line harboring the KRAS G12V mutation.

[0011] FIGS. 6A and 6B show 3D in vitro dose response curves of Compound (A) (FIG. 6A) or Compound (B) (FIG. 6B). FIG. 6C show's a heatmap of 3D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 6D shows a heatmap of 2D in vitro Loewe synergy between Compound(A) and Compound (B). FIGS. 6A-6D were performed in the NCI-H441 NSCLC cell line harboring the KRAS G12V mutation.

[0012] FIGS. 7A and 7B show 2D in vitro dose response curves of Compound (A) (FIG. 7A) or Compound (B) (FIG. 7B). FIG. 7C shows a heatmap of 2D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 7D shows a heatmap of 2D in vitro Loewe synergy between Compound (A) and Compound (B). FIGS. 7A-7D were performed in the SKLU-1 NSCLC cell line harboring the KRAS G12D mutation.

[0013] FIGS. 8A and 8B show 2D in vitro dose response curves of Compound (A) (FIG. 8A) or Compound (B) (FIG. 8B). FIG. 8C shows a heatmap of 2D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 8D shows a heatmap of 2D in vitro Loewe synergy between Compound (A) and Compound (B), FIGS. 8A-8D were performed in the LoVo CRC cell line harboring the KRAS G13D mutation,

[0014] FIGS. 9A and 9B show 3D in vitro dose response curves of Compound (A) (FIG. 9A) or Compound (B) (FIG. 9B). FIG. 9C show's a heatmap of 3D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 9D shows a heatmap of 2D in vitro Loewe synergy between Compound (A) and Compound (B). FIGS. 9A-9D were performed in the LoVo CRC cell line harboring the KRAS G13D mutation.

[0015] FIGS. 10A and 10B show 2D in vitro dose response curves of Compound (A) (FIG. 10A) or Compound (B) (FIG. 10B). FIG. 10C shows a heatmap of 2D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 10D shows a heatmap of 2D in vitro Loewe synergy between Compound (A) and Compound (B). FIGS. 10A-10D were performed in the SW1116 CRC cell line harboring the KRAS G12A mutation.

[0016] FIGS. 11A and 11B show 3D in vitro dose response curves of Compound (A) (FIG. 11 A) or Compound (B) (FIG. 11B). FIG. 11C shows a heatmap of 3D in vitro percent inhibition in response to combination of Compound (A) and Compound (B) at various concentrations. FIG. 11D shows a heatmap of 2D in vitro Loewe synergy between Compound(A) and Compound (B). FIGS. 11A-11D were performed in the SW1116 CRC cell line harboring the KRAS G12A mutation.DETAILED DESCRIPTIONDefinitions

[0017] 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 are 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.

[0018] 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-dihy dr oxy propyl 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, NEL), the nitrogen-based group can be associated with a positive charge (for example, NEL can become NEL”) and the positive charge can be balanced by a negatively charged counterion (such as Ci").

[0019] 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 R-configuration or S-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.

[0020] It is to be understood that where compounds disclosed herein have unfilled valencies, then the valencies are to be filled with hydrogens or isotopes thereof, e.g., hydrogen-1 (protium) and hydrogen-2 (deuterium).

[0021] 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.

[0022] 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 or alcoholates are formed when thesolvent 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.

[0023] 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.

[0024] 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, unrecited 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.

[0025] 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

[0026] Some embodiments described herein relate to the use of a combination of Compound (A) and Compound (B), for treating a cancer. Compound (A) is defined herein asa compound having the structure:(also known as azenosertib or ZN-c3), or a pharmaceutically acceptable salt thereof. Compound (B) is defined.0 o NH, N herein as a compound having the structure:(also known as RMC-6236), or a pharmaceutically acceptable salt thereof. The foregoing definitions of Compound (A) and Compound (B) apply throughout this disclosure. In one embodiment, the cancer being treated is a solid tumor. In one embodiment, the cancer is selected from a colorectal cancer, non-small cell lung cancer and pancreatic cancer.

[0027] Another aspect of this disclosure relates to a method of treating a cancer in a subject comprising administering an effective amount of Compound (A) and an effective amount of Compound (B) as defined above to the subject. In one embodiment, the cancer being treated is a solid tumor. In one embodiment, the cancer is selected from a colorectal cancer, non-small cell lung cancer and pancreatic cancer.

[0028] Yet another aspect of this disclosure relates to a combination of Compound (A) Compound (B) as defined above for use in treating a cancer. The use includes administering to a subject suffering from a cancer a combination of an effective amount of Compound (A) and an effective amount of Compound (B) as defined above. In one embodiment, the cancer being treated is a solid tumor. In one embodiment, the cancer is selected from a colorectal cancer, non-small cell lung cancer and pancreatic cancer.

[0029] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer that is being treated does not have a RAS mutation, i.e., the cancer being treated does not have a NRAS mutation, a HRAS mutation and / or a KRAS mutation. In one embodiment, the cancer does not have a KRAS mutation. The presence of a RAS mutation (e.g., KRAS mutation) can be detected from tumor or plasma specimens from the subject.

[0030] Among the three RAS (KRAS, HRAS, NRAS) protein isoforms, KRAS mutations are believed to be one of the most frequent and prevalent in cancers, including colorectal cancer, pancreatic cancer and non-small cell lung cancer. (See Maitra R, (2021), Therapeutic Approach to KRAS Mutated Colorectal Cancer, Cancer Therapy, MedDocs Publishers. Vol, 4, Chapter 1, pp. 1-5 and J. Luo, Semin Oncol, (2021) 48(1 ): 10- 18). In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer being treated is a RAS-mutated cancer, i.e., the cancer being treated is a NRAS-mutated cancer, a HRAS-mutated cancer and / or a KRAS-mutated cancer. In one embodiment, the cancer being treated is a KRAS-mutated cancer.

[0031] KRAS mutations occur most commonly in codons 12, 13, 59 and / or 61 (including KRAS G12A, G12C, G12D, G12F, G12L, G12R, G12S, G12V, G12Y, G13A, G13C, G13D, G13R, G13S, G13V, A59T, Q61E, Q61H, Q61K, Q61L, Q61P and Q61R), and less common other KRAS codons including codons 117 and / or 146 (including KRAS KI 17N, A146P, A146T or A146V). (See Moore et al., Nat. Rev. Drug Discov. (2020) 19(8): 533-552). In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer that is being treated has at least one KRAS mutation in a codon selected from codons 12, 13, 59 and 61. In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer that is being treated has a G12X mutation. In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer that is being treated has a G13X mutation. In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer that is being treated has a Q61X mutation. In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer that is being treated has at least one KRAS mutation selected from the group consistingof G12A, G12C, G12D, G12F, G12L, G12R, G12S, G12V, G12Y, G13A, G13C, G13D, G13R, G13S, G13V, A59T, Q61E, Q61H, Q61K, Q61L, Q61P and Q61R. In one embodiment, the cancer being treated has a G12A mutation. In one embodiment, the cancer being treated has a G12C mutation. In one embodiment, the cancer being treated has a G12D mutation. In one embodiment, the cancer being treated has a G12F mutation. In one embodiment, the cancer being treated has a G12L mutation. In one embodiment, the cancer being treated has a G12R mutation. In one embodiment, the cancer being treated has a G12S mutation. In one embodiment, the cancer being treated has a G12V mutation. In one embodiment, the cancer being treated has a G12Y mutation. In one embodiment, the cancer being treated has a G13 A mutation. In one embodiment, the cancer being treated has a G13C mutation. In one embodiment, the cancer being treated has a G13D mutation. In one embodiment, the cancer being treated has a G13R mutation. In one embodiment, the cancer being treated has a G13S mutation. In one embodiment, the cancer being treated has a G13V mutation. In one embodiment, the cancer being treated has an A59T mutation. In one embodiment, the cancer being treated has a Q61E mutation. In one embodiment, the cancer being treated has a Q61H mutation. In one embodiment, the cancer being treated has a Q61K mutation. In one embodiment, the cancer being treated has a Q61L mutation. In one embodiment, the cancer being treated has a Q61P mutation. In one embodiment, the cancer being treated has a Q61R mutation.

[0032] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the subject has received (or the cancer has been treated with) at least one (e.g, one, two or three) prior line of therapy, including standard of care (SOC) therapy. In one embodiment, the subject has received (or the cancer has been treated with) one prior line of therapy. In one embodiment, the subject has received (or the cancer has been treated with) two prior lines of therapy. In one embodiment, the subject has received (or the cancer has been treated with) three prior lines of therapy. Such first-, second- or third-line SOC therapies include immunotherapy, such as an immune checkpoint inhibitor; chemotherapy, such as platinum-based chemotherapy, radiation therapy, surgery and anti-angiogenesis therapy (e.g., bevacizumab that is anti -vascular endothelial growth factor or anti-VEGF antibody), each alone or in combination with one another.

[0033] Non-limiting examples of chemotherapy include cisplatin, carboplatin, oxaliplatin, irinotecan, liposomal irinotecan, 5 -fluorouracil (5-FU), leucovorin, paclitaxel, nab-paclitaxel (i.e., a nanoparticle formulation of paclitaxel bound to albumin, while paclitaxel is solvent-based), gemcitabine, pemetrexed including pharmaceutically acceptable salts and derivatives of any of the foregoing, and combinations of any of the foregoing.

[0034] 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 / peinbrolizumab, genolimzumab, nofazinlimab, peinbrolizumab / hyaluronidase, pembrolizumab / quavonlimab, pembrolizumab / vibostolimab, pembrolizumab / quavonlimab, rilvegostomig, sasanlimab, spartalizumab, tebotelimab and volrustoinig (including pharmaceutically acceptable salts of any of the foregoing).

[0035] 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).

[0036] 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 thereof).

[0037] 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).

[0038] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein can be used to treat a subject that has received prior KRAS inhibitor monotherapy, prior KRAS inhibitor combination therapy, a prior combination of KRAS G12C inhibitor monotherapy and / or KRAS G12C combination therapy, e.g, KRAS G12C inhibitor monotherapy, prior KRAS G12C combination therapy, a prior combination of KRAS G12C inhibitor monotherapy and / or KRAS G12C combination therapy. Non-limiting examples of KRAS G12C inhibitors include sotorasib, adagrasib, garsorasib, divarasib (RG-6330), GF-105, opnurasib (JDQ443), olomorasib (LY-3537982),GH-35, glecirasib,F (FMC-376), HS-10370, JMKX-001899, JS- 116, YL-15293, ZG-19018, BEBT-607, BI-1823911, BPI-421286, D3S-001, ERAS-3490,. NoGEC-255, calderasib(MK-1084), (RMC-6291),TEB- 17231,(BBO-8520), LF0001, TSN333, ABSK-071, ADGN-121,ADGN-123, ADGN-531, AFNT-212, APG-1842,(ARS-1620), o (ARS-853),(ASP-2453), AST-NS1902, AU-10458, AU-8653,160, EB-TM1, GRAD-1405, ICP-915, (K20), KP-14,elironrasib (RMC-6291), 880^ (RMC-4998), UCT-00104, VRTX-126, WDB-178, XNW- 14011,(BI-2493), (BI-2865), HYP-2A, and pharmaceutically acceptable salts of any of the foregoing. In one embodiment, the subject has received (or the cancer has been treated with) prior KRAS inhibitor monotherapy, prior KRAS inhibitor combination therapy, a prior combination of KRAS G12C inhibitor monotherapy and / or KRAS G12C combination therapy, e.g., prior KRAS G12C inhibitor monotherapy, prior KRAS G12C combination therapy, a prior combination of KRAS G12C inhibitor monotherapy and / or KRAS G12C combination therapy, wherein the KRAS G12C inhibitor is sotorasib, or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has received (or the cancer has been treated with) prior KRAS inhibitor monotherapy, prior KRAS inhibitor combination therapy, a prior combination of KRAS G12C inhibitor monotherapy and / or KRAS G12C combination therapy, e.g., prior KRAS G12C inhibitor monotherapy, prior KRAS G12C combination therapy, a prior combination of KRAS G12C inhibitor monotherapy and / or KRAS G12C combination therapy, wherein the KRAS G12C inhibitor is adagrasib, or a pharmaceutically acceptable salt thereof.

[0039] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the subject has received (or the cancer has been treated with) prior KRAS G12C monotherapy or prior KR / XS G12C combination therapy (i.e., KRAS G12C inhibitor in combination with a second therapeutic agent, or apharmaceutically acceptable salt thereof, other than Compound (A), or a pharmaceutically acceptable salt thereof). Examples of second therapeutic agents include, but are not limited to, chemotherapy, immune checkpoint inhibitors, along with pharmaceutically acceptable salts thereof, and / or EGFR inhibitors, along with pharmaceutically acceptable salts thereof, (such as cetuximab and panitumumab, etc ), SOS1 inhibitors, along with pharmaceutically acceptable salts thereof, SHP2 inhibitors, along with pharmaceutically acceptable salts thereo.

[0040] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer being treated is a “KRAS G12C inhibitor-sensitive cancer,” which as used herein and per Response Evaluation Criteria in Solid Tumors (RECIST) vl.l refers to a cancer or tumor that is sensitive to KRAS G12C inhibitor therapy that is defined as (1) > 30% decrease from baseline, confirmed at 4 weeks, (2) no > 20% increase over smallest sum observed or (3) no new lesions.

[0041] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer being treated is a KRAS G12C inhibitor-resistant cancer or a KRAS GI2C inhibitor- refractory cancer. In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer being treated is a “KR / XS G12C inhibitor-resistant cancer,” which as used herein and per Response Evaluation Criteria in Solid Tumors (RECIST) vl.l, refers to a cancer or tumor that may have previously responded to, but no longer responds, to KRAS G12C inhibitor therapy that is defined as (1) > 30% decrease from baseline or (2) no > 20% increase over smallest sum observed and no new lesions at 4 weeks. In one embodiment, the cancer acquires resistance, for example, via prior exposure to KRAS G12C inhibitor, either as monotherapy or in combination with other compounds (such as chemotherapy, immune checkpoint inhibitors and EGFR inhibitors such as, but not limited to, cetuximab and panitumumab, SOS1 inhibitors, SHP2 inhibitors, etc., (including pharmaceutically acceptable salts any of the foregoing).

[0042] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer being treated is a “KRAS G12C inhibitor-refractory cancer,” which as used herein and per Response Evaluation Criteria in Solid Tumors (RECIST) vl.l, refers to a cancer or tumor which never responded to KRASG12C inhibitor therapy that is defined as (1) > 30% decrease from baseline or (2) no > 20% increase over smallest sum observed and no new lesions at 4 weeks. In one embodiment, the cancer is inherently resistant.

[0043] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer being treated is colorectal cancer, including, but not limited to, metastatic colorectal cancer, advanced colorectal cancer, KRAS G12X-mutated colorectal cancer, KRAS G13X-mutated colorectal cancer, KRAS Q61X-mutated colorectal cancer, KRAS G12C inhibitor-sensitive colorectal cancer, KRAS G12C inhibitor-resistant colorectal cancer and / or KRAS G12C inhibitor-refractory colorectal cancer,

[0044] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer being treated is pancreatic cancer, including, but not limited to, metastatic pancreatic cancer, advanced pancreatic cancer, KRAS G12X-mutated pancreatic cancer, KRAS G13X-mutated pancreatic cancer, KRAS Q61X-mutated colorectal cancer, KRAS G12C inhibitor-sensitive pancreatic cancer, KRAS G12C inhibitor-resistant pancreatic cancer and / or KRAS G12C inhibitor-refractory pancreatic cancer.

[0045] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein, the cancer being treated the cancer is non-small cell lung cancer, including, but not limited to, metastatic non-small cell lung cancer, advanced pancreatic cancer, KRAS G12X-mutated non-small cell lung cancer, KRAS G13X-mutated non-small cell lung cancer, KRAS Q61X-mutated non-small cell lung cancer, KRAS G12C inhibitor-sensitive non-small cell lung cancer, KRAS G12C inhibitor-resistant non-small cell lung cancer and / or KRAS G12C inhibitor-refractory non-small cell lung cancer.

[0046] When the treatment is a combination of compounds, the order of administration or use of the combination of Compound (A) and Compound (B) as defined herein can vary. In some embodiments, Compound (A) and Compound (B) are administered or used sequentially. In some embodiments, Compound (A) can be administered prior to Compound (B). In other embodiments, Compound (A) can be administered subsequent to or after Compound (B). In still other embodiments, Compound (A) can be administered concomitantly or concurrently with Compound (B).

[0047] In some embodiments of the uses and methods related to the combination of Compound (A) and Compound (B) provided herein can decrease the number and / or severity of side effects that can be attributed to monotherapy of the Compound (B).

[0048] Using a combination of Compound (A) and Compound (B) 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.

[0049] 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.

[0050] 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) is administered as a monotherapy. For example, the amount of Compound (B) used in a combination described herein can be less compared to the amount of Compound (B) needed to achieve the same reduction in a disease marker (for example, tumor size) when 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 (B) is 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

[0051] Compound (A) can be provided in a pharmaceutical composition. Likewise, Compound (B) can be provided in a pharmaceutical composition.

[0052] 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.

[0053] As used herein, a “carrier” refers to a compound that facilitates the incorporation of a compound into cells or tissues. For example, without limitation, dimethyl sulfoxide (DMSO) is a commonly utilized carrier that facilitates the uptake of many organic compounds into cells or tissues of a subject.

[0054] As used herein, a “diluent” refers to an ingredient in a pharmaceutical composition that lacks appreciable pharmacological activity but may be pharmaceutically necessary or desirable. For example, a diluent may be used to increase the bulk of a potent drug whose mass is too small for manufacture and / or administration. It may also be a liquid for the dissolution of a drug to be administered by injection, ingestion or inhalation. A common form of diluent in the art is a buffered aqueous solution such as, without limitation, phosphate buffered saline that mimics the pH and isotonicity of human blood.

[0055] As used herein, an “excipient” refers to an essentially inert substance that is added to a pharmaceutical composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability etc., to the composition. For example, stabilizers such as anti-oxidants and metal-chelating agents are excipients. In an embodiment, the pharmaceutical composition comprises an anti-oxidant and / or a metalchelating agent. A “diluent” is a type of excipient.

[0056] In some embodiments, Compound (B) can be provided in a pharmaceutical composition that includes Compound (A). In other embodiments, Compound (B) can beadministered in a pharmaceutical composition that is separate from a pharmaceutical composition that includes Compound (A).

[0057] The pharmaceutical compositions described herein can be administered to a human patient per se, or in pharmaceutical compositions where they are mixed with other 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.

[0058] 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 m 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.

[0059] 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 some embodiments, Compound (A) can be administered orally. In some embodiments, Compound (A) can be provided to a subject by the same route of administration as Compound (B). In other embodiments, Compound (A) can be provided to a subject by a different route of administration as Compound (B).

[0060] One may also administer the compound, salt and / or composition in a local rather than systemic manner, for example, via injection or implantation of the compound directly into the affected area, often in a depot or sustained release formulation. Furthermore, one may administer the compound in a targeted drug delivery system, for example, in a liposome coated with a tissue-specific antibody. The liposomes will be targeted to and taken up selectively by the organ. For example, intranasal or pulmonary’ delivery to target a respiratory disease or condition may be desirable.

[0061] The compositions may, if desired, be presented in a pack or dispenser device w’hich may contain one or more unit dosage forms containing the active ingredient. The packmay for example comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, may be the labeling approved by the U. S. Food and Drug Administration for prescription drugs, or the approved product insert. Compositions that can include a compound and / or salt described herein formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.Uses and Methods of Treatment

[0062] As provided herein, in some embodiments, a combination of compounds that includes an effective amount of Compound (A) and an effective amount of Compound (B) can be used to treat a cancer.

[0063] 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, the cancer can be a recurrent cancer.

[0064] As used herein, a “subject” refers to an animal that is the object of treatment, observation or experiment. “Animal” includes cold- and warm-blooded vertebrates and invertebrates such as fish, shellfish, reptiles and, m particular, mammals. “Mammal” includes, without limitation, mice, rats, rabbits, guinea pigs, dogs, cats, sheep, goats, cows, horses, primates, such as monkeys, chimpanzees, and apes, and, in particular, humans. In some embodiments, the subject can be human. In some embodiments, the subject can be a child and / or an infant. In other embodiments, the subject can be an adult.

[0065] 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.

[0066] The term “effective amount” is used to indicate an amount of an active compound, or pharmaceutical agent, that elicits the biological or medicinal response indicated. 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.

[0067] 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.

[0068] 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.

[0069] As will be readily apparent to one skilled in the art, the useful in vivo dosage to be administered and the particular mode of administration will vary depending upon the age, weight, the severity of the affliction, the mammalian species treated, the particular compounds employed and the specific use for which these compounds are employed. The determination of effective dosage levels, that is the dosage levels necessary to achieve the desired result, canbe accomplished by one skilled in the art using routine methods, for example, human clinical trials, in vivo studies and in vitro studies. For example, useful dosages of Compounds (A) and Compound (B) can be determined by comparing their in vitro activity, and in vivo activity in animal models. Such comparison can be done by comparison against an established drug, such as cisplatin and / or gemcitabine.

[0070] Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety which are sufficient to maintain the modulating effects, or minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from in vivo and / or in vitro data. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. However, HPLC assays or bioassays can be used to determine plasma concentrations. Dosage intervals can also be determined using MEC value. Compositions should be administered using a regimen which maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90% and most preferably between 50-90%. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

[0071] 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 according to the age, body weight and response of the individual patient. A program comparable to that discussed above may be used in veterinary medicine.

[0072] 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 ofparticular 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

[0073] 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.Example 1: Combination of Compound (A) and Compound (B) Demonstrates Synergy in NSCLC Model Harboring the KRAS G12C Mutation In Vitro in 2D and 3D Cellular AssaysIn Vitro 2D Cell Titer Gio (CTG) Assays

[0074] Compound (A) and Compound (B) in DMSO were deposited into flat bottom opaque white 96- well plates using an automated drug dispenser at the indicated concentrations. Total DMSO content was normalized to 0.1% of the total volume in all assay conditions. Cell suspensions containing 1.5 x 103total cells in 100 μL culture media were deposited in each well of the 96 well plate containing compound. Plates were incubated at 37 °C with 95% oxygen and 5% CO₂ for 72 h. After 72 h, plates were removed from the incubator and allowed to come to room temperature. Room temperature CTG reagent (Promega, Cat# G7571) was added to each well at 1:1 ratio of culture media to CTG reagent. Plates were agitated at 520 revolutions per minute (rpm) for 2 mins, allowed to stabilize protected from light for 10 mins, then luminescence was measured on an M5e plate reader (SpectraMax). Percent viability is calculated as percentage of cell viability relative to DMSO-only vehicle control. Percent viability was input into the SynergyFinder web tool (https: / / www.synergyfinderplus.org / ). Dose response curves, percent viability heatmaps, and synergy heatmaps were downloaded via PDF from the SynergyFinder website. The Loewe synergy score was determined for each combination. Loewe values > 10 indicate synergisticcombination effects of the drugs, Loewe values from -10 to 10 indicate additive effects, and Loewe values < -10 indicate antagonism.In Vitro 3D Spheroid Growth Assays

[0075] Cell suspensions containing 1.5 x 103total cells in 90 μL culture media were deposited in each well of an ultra-low attachment 96-well plate. Plates were incubated at 37 °C with 95% oxygen and 5% CO2 for 24 h to allow’ for spheroid formation. After 24 h, Compound (A) and Compound (B) in DMSO were deposited into each plate using an automated drug dispenser at the indicated concentrations. Total DMSO content was normalized to 0.1% of the total volume in all assay conditions. Plates were incubated at 37 °C with 95% oxygen and 5% CO2 for 168 h. After 72 h, plates were removed from the incubator and allowed to come to room temperature. Room temperature 3D CTG reagent (Promega, Cat# G9683) was added to each well at 1: 1 ratio of culture media to 3D CTG reagent. Plates were agitated at 520 revolutions per minute (rpm) for 5 mins, allowed to stabilize protected from light for 10 mins, then luminescence was measured on an M5e plate reader (SpectraMax). Percent viability calculations, generation of dose response curves, percent viability heatmaps, and synergy heatmaps, and determination of Loewe synergy scores were carried out as described above.

[0076] FIGS. 1A-1D and FIGS. 2A-2D depict the NCI-H1792 NSCLC cell line harboring the KRAS G12C mutation given escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIGS. 1A-1D) and 3D (FIGS. 2A-2D). FIGS. 1A and 2A each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (A) and indicate that as Compound (A) dose increases, percent inhibition of 2D (FIG. 1A) and 3D (FIG. 2A) cell growth also increases, demonstrating sensitivity of NCI-H1792 cells to Compound (A) as a monotherapy.

[0077] FIGS. 1B and 2B each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (B) and indicate that as Compound (B) dose increases, percent inhibition of cell growth also increases, demonstrating sensitivity of NCI-H1792 cells to Compound (B) as a monotherapy in 2D (FIG. 1B) and 3D (FIG. 2B).

[0078] FIGS. 1C and 2C each depict a heatmap generated from the SynergyFinder tool of percent inhibition in response to escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIG. 1C) and 3D (FIG. 2C). Percent inhibition scores demonstrate increased percent inhibition of NCI-H1792 cells for the combination of Compound (A) and Compound (B) when compared against monotherapy of either compound at the same concentration.

[0079] FIGS. ID and 2D each depict a heatmap generated from the SynergyFinder tool of the Loewe synergy score for each combination. (James et al. Cell 172, 578-589.e17 (2018)) have demonstrated that R AS-driven models grown as 3D spheroids may more accurately reflect the sensitivity to KRAS inhibition in vivo. Synergy Score > 10 is considered synergistic. The data indicates that in the NCI-H1792 cell line combination treatment with Compound (A) combined with Compound (B) produced synergy scores > 10 in at least one dose combination level, indicating that this combination is synergistic in 2D (FIG. 1D) and 3D (FIG. 2D) in vitro.Example 2: Combination of Compound (A) and Compound (B) Demonstrates Synergy in CRC Model Harboring the KRAS G12V Mutation In Vitro in 2D and 3D Cellular Assays

[0080] FIGS. 3A-3D and FIGS.4A-4D depict the SW403 CRC cell line harboring the KRAS G12V mutation given escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIGS. 3A-3D) and 3D (FIGS. 4A-4D). FIGS. 3A and 4A each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (A) and indicate that as Compound (A) dose increases, percent inhibition of 2D (FIG. 3A) and 3D (FIG. 4A) cell growth also increases, demonstrating sensitivity of SW403 cells to Compound (A) as a monotherapy.

[0081] FIGS. 3B and 4B each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (B) and indicate that as Compound (B) dose increases, percent inhibition of cell growth also increases, demonstrating sensitivity of SW403 cells to Compound (B) as a monotherapy in 2D (FIG. 3B) and 3D (FIG.4B).

[0082] FIGS. 3C and 4C each depict a heatmap generated from the SynergyFinder tool of percent inhibition in response to escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIG. 3C) and 3D (FIG. 4C). Percent inhibition scores demonstrate >50 percent inhibition of SW403 cells at low dose for the combination of Compound (A) and Compound (B) when compared against monotherapy of either compound at the same concentration, demonstrating strong potency with a minimal dose.

[0083] FIGS. 3D and 4D each depict a heatmap generated from the SynergyFinder tool of the Loewe synergy score for each combination. Synergy Score > 10 is considered synergistic. The data indicates that in the SW403 cell line combination treatment with Compound (A) combined with Compound (B) produced synergy scores > 10 in at low dose combination level, indicating that this combination is synergistic in 2D (FIG. 3D) and 3D (FIG. 4D) in vitro.Example 3: Combination of Compound (A) and Compound (B) Demonstrates Synergy in NSCLC Model Harboring the KRAS G12V Mutation In Vitro in 2D and 3D Cellular Assays

[0084] FIGS. 5A-5D and FIGS. 6A-6D depict the NCI-H441 NSCLC cell line harboring the KRAS G12V mutation given escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIGS. 5A-5D) and 3D (FIGS. 6A-6D). FIGS. 5A and 6A each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (A) and indicate that as Compound (A) dose increases, percent inhibition of 2D (FIG. 5A) and 3D (FIG. 6A) cell growth also increases, demonstrating sensitivity of NCI-H441 cells to Compound (A) as a monotherapy.

[0085] FIGS. 5B and 6B each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (B) and indicate that as Compound (B) dose increases, percent inhibition of cell growth also increases, demonstrating sensitivity of NCI-H441 cells to Compound (B) as a monotherapy in 2D (FIG. 5B) and 3D (FIG. 6B)

[0086] FIGS. 5C and 6C each depict a heatmap generated from the SynergyFinder tool of percent inhibition in response to escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIG. 5C) and 3D (FIG. 6C). Percent inhibition scores demonstrate increased percent inhibition across a wide range of doses of NCI-H441 cells for the combination of Compound (A) and Compound (B) when compared against monotherapy of either compound at the same concentration.

[0087] FIGS. 5D and 6D each depict a heatmap generated from the SynergyFinder tool of the Loewe synergy score for each combination. Synergy Score > 10 is considered synergistic. The data indicates that in the NCI-H441 cell line combination treatment with Compound (A) combined with Compound (B) produced synergy scores > 20 in multiple dose combination levels, indicating that this combination is highly synergistic in 2D (FIG. 5D) and 3D (FIG. 6D) in vitro.Example 4: Combination of Compound ( A) and Compound (B) Demonstrates Synergy in NSCLC Model Harboring the KRAS G12D Mutation In Vitro in 2D and 3D Cellular Assays

[0088] FIGS. 7A-7D depict the SKLU-1 NSCLC cell line harboring the KRAS G12D mutation given escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIGS. 7A-7D). FIG. 7 A depicts a dose response curve generated from the SynergyFinder tool m response to escalating doses of Compound (A) and indicate that as Compound (A) dose increases, percent inhibition of 2D (FIG. 7A) cell growth also increases, demonstrating sensitivity of SKLU-1 cells to Compound (A) as a monotherapy.

[0089] FIG. 7B depicts a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (B) and indicate that as Compound (B) dose increases, percent inhibition of cell growth also increases, demonstrating sensitivity of SKLU-1 cells to Compound (B) as a monotherapy in 2D (FIG. 7B).

[0090] FIG. 7C depicts a heatmap generated from the SynergyFinder tool of percent inhibition in response to escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIG. 7C). Percent inhibition scores demonstrate increased percent inhibition of SKLU-1 cells for thecombination of Compound (A) and Compound (B) at both low and high doses when compared against monotherapy of either compound at the same concentration.

[0091] FIG. 7D depicts a heatmap generated from the SynergyFinder tool of the Loewe synergy score for each combination. Synergy Score > 10 is considered synergistic. The data indicates that in the SKLU-1 cell line combination treatment with Compound (A) combined with Compound (B) produced synergy scores > 20 in multiple dose combination levels at high and low dose, indicating that this combination is synergistic in 2D (FIG. 7D) in vitro.Example 5: Combination of Compound ( A) and Compound (B) Demonstrates Synergy in LoVo Model Harboring the KRAS G13D Mutation In Vitro in 2D and 3D Cellular Assays

[0092] FIGS. 8A-8D and FIGS. 9A-9D depict the LoVo CRC cell line harboring the KRAS G13D mutation given escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIGS. 8A-8D) and 3D (FIGS.9A-9D). FIGS. 8A and 9A each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (A) and indicate that as Compound (A) dose increases, percent inhibition of 2D (FIG. 8A) and 3D (FIG. 9A) cell growth also increases, demonstrating sensitivity of LoVo cells to Compound (A) as a monotherapy.

[0093] FIGS. 8B and 9B each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (B) and indicate that as Compound (B) dose increases, percent inhibition of cell growth also increases, demonstrating sensitivity of LoVo cells to Compound (B) as a monotherapy in 2D (FIG. 8B) and 3D (FIG.9B)

[0094] FIGS. 8C and 9C each depict a heatmap generated from the SynergyFinder tool of percent inhibition in response to escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIG. 8C) and 3D (FIG. 9C). Percent inhibition scores demonstrate increased percent inhibition of LoVo cells for the combination of Compound (A) and Compound (B) when compared against monotherapy of either compound at the same concentration.

[0095] FIGS. 8D and 9D each depict a heatmap generated from the SynergyFinder tool of the Loewe synergy score for each combination. Synergy Score > 10 is considered synergistic. The data indicates that in the LoVo cell line combination treatment with Compound (A) combined with Compound (B) produced synergy scores > 10 in at least one dose combination level, indicating that this combination is synergistic in 2D (FIG. 8D) and 3D (FIG. 9D) in vitro.Example 6: Combination of Compound (A) and Compound (B) Demonstrates Synergy in CRC Model Harboring the KRAS G12A Mutation In Vitro in 2D and 3D Cellular Assays

[0096] FIGS. 10A-10D and FIGS. 11A-11D depict the SW1116 CRC cell line harboring the KRAS G12A mutation given escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIGS. 10A-10D) and 3D (FIGS. 11A-11D). FIGS. 10A and 11A each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (A) and indicate that as Compound (A) dose increases, percent inhibition of 2D (FIG. 10A) and 3D (FIG. 11A) cell growth also increases, demonstrating sensitivity of NCI-H1792 cells to Compound (A) as a monotherapy.

[0097] FIGS. 10B and 11B each depict a dose response curve generated from the SynergyFinder tool in response to escalating doses of Compound (B) and indicate that as Compound (B) dose increases, percent inhibition of cell growth also increases, demonstrating sensitivity of SW1116 cells to Compound (B) as a monotherapy in 2D (FIG. 10B) and 3D (FIG. 11B)

[0098] FIGS. 10C and 11C each depict a heatmap generated from the SynergyFinder tool of percent inhibition in response to escalating doses of Compound (A) as a single agent or combined with Compound (B) in escalating doses in a matrixed format in 2D (FIG. 10C) and 3D (FIG. 11C). Percent inhibition scores demonstrate increased percent inhibition of SW1116 cells for the combination of Compound (A) and Compound (B) when compared against monotherapy of either compound at the same concentration.

[0099] FIGS. 10D and 11D each depict a heatmap generated from the SynergyFinder tool of the Loewe synergy score for each combination. Synergy Score > 10 is considered synergistic. The data indicates that in the SW1116 cell line combination treatmentwith Compound (A) combined with Compound (B) produced synergy scores > 10 in at least one dose combination level in 2D and produced synergy scores > 20 in at least one dose combination level, indicating that this combination is synergistic in 2D (FIG. 10D) and highly synergistic in 3D (FIG. 11D) in vitro.

[0100] Altogether, the data in the foregoing Examples 1-6 show that the combination of Compound (A) and Compound (B) is consistently generally superior to Compound (B) monotherapy across a range of exemplary cancer models, including, but not limited to, NSCLC and CRC models carrying different mutations.

[0101] 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.

Claims

WHAT IS CLAIMED IS:

1. Use of a combination of Compound (A) and Compound (B) in the preparation of a medicament for treating a cancer, wherein:Compound (A) is a pharmaceutically acceptable salt thereof; andCompound (B) is / , or a pharmaceutically acceptable salt thereof.

2. The use of claim 1, wherein Compound (A), or a pharmaceutically acceptable salt thereof, is administered prior to the administration of Compound (B), or a pharmaceutically acceptable salt thereof.

3. The use of claim 1, wherein Compound (A), or a pharmaceutically acceptable salt thereof, is administered subsequent to the administration of Compound (B), or a pharmaceutically acceptable salt thereof.

4. The use of claim 1, wherein Compound (A), or a pharmaceutically acceptable salt thereof, is administered concomitantly with Compound (B), or a pharmaceutically acceptable salt thereof,5. The use of claim 4, wherein Compound (A), or pharmaceutically acceptable salts thereof, are provided in a composition that includes Compound (B), or a pharmaceutically acceptable salt thereof.

6. The use of any one of claims 1 to 4, wherein Compound (A), or pharmaceutically acceptable salts thereof, is provided in a composition that is separate from Compound (B), or a pharmaceutically acceptable salt thereof.

7. The use of any one of claims 1 to 6, wherein the cancer does not have a RAS mutation or KRAS mutation.

8. The use of any one of claims 1 to 6, wherein the cancer is a RAS-mutated cancer.

9. The use of any one of claim 1 to 6 or claim 8, wherein the cancer is a KRAS-mutated cancer,10. The use of any one of claims 1 to 6, 8 and 9, wherein the cancer has at least one KRAS mutation in a codon selected from codons 12, 13, 59 and 61.

11. The use of any one of claims 1 to 6 or 8 to 10, wherein the cancer has a G12X mutation,12. The use of any one of claims 1 to 6 or 8 to 11, w'herein the cancer has a G13X mutation.

13. The use of any one of claims 1 to 6 or 8 to 12, wherein the cancer has a Q61X mutation.

14. The use of any one of claims 1 to 6 or 8 to 13, wherein the cancer has at least one KRAS mutation selected from the group consisting of G12A, G12C, G12D, G12F, G12L, G12R, G12S, G12V, G12Y, G13A, G13C, G13D, G13R, G13S, G13V, A59T, Q61E, Q61H, Q61K, Q61L, Q61P and Q61R.

15. The use of any one of claims 1 to 14, wherein the cancer is selected from a colorectal cancer, a non-small cell lung cancer and a pancreatic cancer.

16. The use of any one of claims 1 to 15, wherein the cancer is a colorectal cancer.

17. The use of any one of claims 1 to 15, wherein the cancer is a non-small cell lung cancer.

18. The use of any one of claims 1 to 15, wherein the cancer is a pancreatic cancer.

19. The use of claim 18, wherein the pancreatic cancer is pancreatic ductal ad enocarcinoma.

20. The use of any one of claims 1 to 19, wherein the cancer has been treated with at least one prior line of therapy.

21. The use of claim 20, wherein the at least one prior line of therapy comprises chemotherapy, immunotherapy or a combination thereof.

Images