Wee1 inhibitor combination therapy

Abstract

Disclosed herein is use of a combination of Compound (A) (azenosertib), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, such as sotorasib or adagrasib, or a pharmaceutically acceptable salt thereof, for treating a disease or condition, such as a colorectal, pancreatic and / or non-small cell lung cancer.

Description

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, for example, in the Application Data Sheet or Request as filed with the present application, are hereby incorporated by reference under 37 CFR 1.57, and Rules 4.18 and 20.6, including U.S. Provisional Application Nos. 63 / 560,413, filed March 1, 2024, 63 / 566,801, filed March 18, 2024, 63 / 699,385, filed September 26, 2024 and 63 / 733,576, filed December 13, 2024, each of which is incorporated by reference in their entireties.BACKGROUNDField

[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] KRAS G12C (z.f., a mutant form of the K-Ras protein carrying the G12C mutation) is a potent oncogenic driver which results in downstream hyperactivation of MAPK signaling and unchecked oncogenic growth, while simultaneously increasing replication stress (RS) and accumulation of DNA damage. KRAS G12C-specific inhibitors are approved and / or recommended for treatment of metastatic non-small cell lung cancer (NSCLC) and colorectal cancer (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

[0004] Some embodiments described herein relate to the use of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer in a subject. Compound (A) ispharmaceutically acceptable salt thereof. The cancer is a KRAS G12C inhibitor-sensitive cancer, a KRAS G12C inhibitor-resistant cancer or a KRAS G12C inhibitor-refractory cancer, and is selected from colorectal cancer, pancreatic cancer and non-small cell lung cancer.

[0005] In the accompanying Examples, the efficacy and synergistic effect of combining Compound (A) with two exemplary FDA-approved KRAS G12C inhibitors across a panel of cell lines and xenograft models representing tumor indications where KRAS G12C alterations are common are provided. As shown by the in vitro in 2D anchorage-dependent (FIGS. 1A, IB and IE) and 3D anchorage-independent (FIGS. 2A-2C) models, along with the in vivo xenograft models (FIGS. 3A-3C, 4A-4F, 5A-5C and 6A-6D) of NSCLC, CRC and PDAC, Compound (A) shows both meaningful monotherapy potency and significant synergy with both sotorasib and adagrasib.

[0006] In cell-line derived xenograft (CDX) models of NSCLC, CRC and PDAC that are already relatively sensitive to KRAS G12C inhibition, Compound (A) demonstrates not only has statistically significant activity on its own, but also in combination with KRAS G12C inhibitors. The combination not only greatly improves anti-tumor efficacy, but often resulted in meaningful tumor regression (FIGS. 3A-3C, 4A-4F and 5A-C). A key clinical challenge of KRAS G12C inhibitor monotherapy is the relatively short progression free survival (PFS). The in vivo data provided herein (FIGS. 5A-5F) suggests that combining Compound (A) with a KRAS G12C inhibitor may enhance the efficacy of KRAS G12C inhibitors and ultimately extend PFS in subjects with KRAS G12C NSCLC.

[0007] Although monotherapy with KRAS G12C inhibitors have demonstrated clear clinical benefit in subjects with KRAS G12C mutations, a substantial portion of thesesubjects with KRAS G12C mutations do not respond. In addition to subjects that are inherently less responsive to KRAS G12C monotherapy, even those subjects that initially respond to treatment eventually relapse. As provided herein, intrinsically resistant and acquired- resistance models have been shown to be sensitive to Compound (A) and display synergistic TGI when combined with KRAS G12C inhibitors (FIGS. 6A-6D).BRIEF DESCRIPTION OF THE DRAWINGS

[0008] FIGS. 1A and IB show 7-day combination treatment dose matrices in KRAS G12C NSCLC cell lines cultured in 2D by Loewe synergy model (FIG. 1A) or Bliss Independence model (FIG. IB). FIG. 1C shows Western blot of protein expression from NCL H2122 NSCLC cells treated with DMSO, 250 nM of Compound (A), 62.5 nM of sotorasib, 25 nM of adagrasib or a combination for 24 hours. FIG. ID shows the DNA content analysis by flow cytometry represented as percentage of total cells in each cell cycle phase. EdU+ and EdU- DNA distinguish active versus stalled DNA replication. NCLH2122 NSCLC cells were treated with DMSO, 500 nM of Compound (A), 62.5 nM of sotorasib, 50 nM of adagrasib or the combinations for 24 hours. FIG. IE shows quantification of yH2AX and c-Casp-3 / 7 protein levels from cell lines treated for 24 hours based on doses associated with the highest calculated Loewe synergy score. FIG. IF shows 7-day combination treatment dose matrices in MIA PaCa-2 PDAC, SW837 CRC, and NCI-H2122 NSCLC cell lines cultured in 2D. FIG. 1G shows Western blot of protein expression from MIA PaCa-2 PDAC cells treated with DMSO, 310 nM of Compound (A), 12.5 nM of sotorasib or a combination, and SW837 CRC cells treated with DMSO, 200 nM of Compound (A), 10 nM of sotorasib or a combination for 24 hours. FIG. IH shows 7-day combination treatment dose matrices in MIA PaCa-2 PDAC, SW837 CRC, and NCLH2122 NSCLC cell lines cultured in 2D after treatment with siRNA control or siRNA against WEE1. FIG. II shows Western blot of protein expression from indicated cell lines treated with 100 nM siControl or siWEEl for 48 hours.

[0009] FIG. 2A shows 7-day combination treatment dose matrices in KRAS G12C NSCLC cell lines cultured as 3D spheroids. FIGS. 2B and 2C show longitudinal cell growth analysis of NCLH23 spheroids treated with DMSO, 125 nM of Compound (A), 5 nM of sotorasib, 5 nM of adagrasib or a combination for 7 days. FIG. 2D shows Western blot of protein expression from NCLH23 spheroids treated with DMSO, 260 nM of Compound (A),of yH2AX and c-Casp-3 / 7 fold change in protein levels in response to treatment relative to DMSO.

[0010] FIG. 3A shows mean tumor volume ± SEM of subcutaneous NCI-H2122 xenografts in NOD / SCID mice treated for 25 days (n=9 / group). FIG. 3B shows percent change in tumor volume (ATV) of individual mice on day 25 of treatment. FIG. 3C shows mean percent change in body weight relative to day 0 (ABW) ± SEM. NCI-H2122 was noted to be cachexic in NOD / SCID mice (seen by minor weight loss in vehicle group), but treatments were well-tolerated and did not exacerbate weight loss relative to vehicle. The bottom dashed line indicates -15% cutoff in ABW. FIG. 3D shows Western blot of protein expression from NCI-H2122 tumors treated with one dose of the indicated compound(s) and also quantification of yH2AX and c-Casp-3 / 7 fold change in protein levels in response to treatment relative to DMSO. FIG. 3E shows Western blot of protein expression from SW1573 cells treated with DMSO, 625 nM of Compound (A), 1.25 pM adagrasib, or a combination of both for 4 h or 24 h. FIG. 3F shows Western blot of protein expression from SW1573 tumors treated with five daily doses of the indicated compounds and collected either 4 h or 24 h after the last dose.

[0011] FIG. 3G shows representative images at 40x magnification of Ki67 IHC performed on NCI-H2122 NSCLC tumors treated with a single dose of Compound (A), sotorasib, or the combination. FIG. 3H shows the modified Ki67 H-scores of NCI-H2122 tumors treated with one dose of the indicated compounds or the combination thereof. * = p < 0.05; ** = p < 0.001; *** p < 0.0001. FIG. 31 shows representative images of H&E staining at 1 x, 20x or 40x magnification performed on NCI-H2122 tumors treated with one dose of the indicated compounds or the combination, where the yellow circles indicate regions of cell death. FIG. 3J is a graph showing the cell death of NCI-H2122 tumors treated with one dose of the indicated compounds as scored by a board-certified veterinary pathologist with experience in laboratory animals and toxicologic pathology. ** = p < 0.001.

[0012] FIGS. 4A and 4D respectively show percent change in tumor volume (ATV) and mean tumor volume ± SEM of individual SW837 xenografts in NOD / SCID mice on day 21 of treatment (n=8 / group). FIGS. 4B and 4E respectively show ATV and mean tumor volume ± SEM of individual SW1463 xenografts in NCG mice on day 21 of treatment(n=8 / group). FIGS. 4C and 4F respectively show ATV and mean tumor volume ± SEM of individual MIA PaCa-2 xenografts in BALB / c nude mice on day 21 of treatment (n=8 / group).

[0013] FIGS. 4G-4N show mean percent change in body weight from day 0 of SW837, SW1463, MIA PaCa-2, NCI-H2030, NCI-H1792, SW1537, CR2528 and NCI- H2030-R xenografts treated for a range of 21-67 days for different models (n=8 / group or n=9 / group).

[0014] FIG. 5A shows mean tumor volume ± SEM of subcutaneous NCI-H2030 xenografts in NOD / SCID mice (n=9 / group). FIG. 5B shows mean tumor volume ± SEM of subcutaneous NCI-H1792 xenografts in NOD / SCID mice (n=9 / group). FIG. 5C shows individual animal tumor volumes on day 71 or day 102 for adagrasib monotherapy and combinations from FIG. 5B. FIG. 5D is a Kaplan-Meier survival curve that shows the probability of tumor regrowth defined as the time until tumor volume reaches > 250 mm3. (NR = Not Reached). FIGS. 5E and 5F are Kaplan-Meier survival curves that show the probability of tumor regrowth defined as the time until tumor volume reaches > 100 mm3, at low (FIG. 5E) and high (FIG. 5F) dose of adagrasib.

[0015] FIG. 6A shows mean tumor volume ± SEM of subcutaneous SW1537 xenografts in NOD / SCID mice (n = 9 / group). FIG. 6B shows mean tumor volume ± SEM of subcutaneous CR2528 PDX in BALB / c nude mice treated for 24 days (n=8 / group). FIG. 6C shows mean tumor volume ± SEM of subcutaneous NCI-H2030-R xenografts in NOD / SCID mice (n=9 / group). FIG. 6D shows the ratio of tumor volume in treated versus control mice (% T / C) of subcutaneous PDX models implanted into athymic nude (Crl:NU(NCr)-Foxnlnu) mice (n = 3 / group).DETAILED DESCRIPTIONDefinitions

[0016] 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.

[0017] 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)m ethylamine, 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 be associated with a positive charge (for example, NH2 can become NHs ) and the positive charge can be balanced by a negatively charged counterion (such as Cl’).

[0018] 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.

[0019] 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).

[0020] 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.

[0021] 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 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.

[0022] 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.

[0023] 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’ shouldbe 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.

[0024] 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

[0025] Some embodiments described herein relate to the use of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer in a subject. Compound (A) is(also known as azenosertib or ZN-c3), or a pharmaceutically acceptable salt thereof. The cancer is a KRAS G12C inhibitor-sensitivecancer, a KRAS G12C inhibitor-resistant cancer or a KRAS G12C inhibitor-refractory cancer, and is selected from colorectal cancer, pancreatic cancer and non-small cell lung cancer.

[0026] 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, or a pharmaceutically acceptable salt thereof, and an effective amount of a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, to the subject. The cancer is a KRAS G12C inhibitor-sensitive cancer, a KRAS G12C inhibitor-resistant cancer or a KRAS G12C inhibitor-refractory cancer, and is selected from colorectal cancer, pancreatic cancer and non- small cell lung cancer.

[0027] Yet another aspect of this disclosure relates to a combination of Compound(A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, 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, or a pharmaceutically acceptable salt thereof, and an effective amount of a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof. The cancer is a KRAS G12C inhibitor-sensitive cancer, a KRAS G12C inhibitorresistant cancer or a KRAS G12C inhibitor-refractory cancer, and is selected from colorectal cancer, pancreatic cancer and non-small cell lung cancer.

[0028] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the subject has a KRAS G12C mutation. The presence of a KRAS G12C mutation can be detected from tumor or plasma specimens from the subject. 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). 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, GBR, G13S, G13V, A59T, Q61E, Q61H, Q61K, Q61L, Q61P and Q61R), andless common other KRAS codons including codons 1 17 and / or 146 (including KRAS KI 17N, A146P, A146T or A146V). (See Moore et al., Nat. Rev. Drug Discov. (2020) 19(8):533-552).

[0029] The first success in directly targeting KRAS grew out of the unique biochemistry of the codon 12 glycine-to-cysteine (G12C) mutation. Cysteine thiols are uniquely nucleophilic amongst amino acid side chains. This is seen in formation of disulfide bonds and made the cysteine residue in mutant KRAS G12C an attractive pharmacologic target. This property enabled disulfide-fragment-based techniques to screen hundreds of potential small-molecule targets of G12C-mutant KRAS in a mutant-specific manner, sparing WT KRAS. Building on the fact that G12C was targetable, two clinical compounds have now obtained approval: sotorasib and adagrasib.

[0030] As defined herein, a “KRAS G12C inhibitor” is a molecule, which may be an organic chemistry small molecule, a nucleic acid-based molecule (e. , siRNA, shRNA, anti-sense oligonucleotide or ASO, etc.), and / or an antibody or an antigen-binding fragment thereof (including pharmaceutically acceptable salts of any of the foregoing), that specifically or non-specifically targets and binds to the KRAS G12C mutant protein and subsequently at least partially (z.e., at least 5%) inhibits or inactivates the protein. The KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, may simultaneously targets, for example, other mutant forms of KRAS (e.g., G12D and G12V) as well as other proteins including, but not limited to, NRAS G12C, PI3K-alpha, p53, S0S1, Ras, PARP, and / or CD3. Non-limiting examples of G12C inhibitors include sotorasib, adagrasib, garsorasib, divarasib, GF-105, JDQ443, LY-3537982, GH-35, glecirasib, FMC-376, HS-10370, JMKX-001899, JS-116, YL- 15293, ZG-19018, BEBT-607, BI-1823911, BPI-421286, D3S-001, ERAS-3490, GEC-255, MK-1084, RG-6330, RMC-6291, TEB-17231, BBO-8520, LF0001, TSN333, ABSK-071, ADGN-121, ADGN-122, ADGN-123, ADGN-531, AFNT-212, APG-1842, ARS-1620, ARS- 853, ASP-2453, AST-NS1902, AU-10458, AU-8653, AZD-4625, AZD-4747, BPI-2361, EB- 160, EB-TM1, GRAD-1405, ICP-915, K20, KP-14, LC-2, MRTX-1257, R023, RM-007, RM- 018, RM-032, UCT-00104, VRTX-126, WDB-178, XNW-14011, YF135, BI-2493, BI-2865, HYP-2A, and pharmaceutically acceptable salts of any of the foregoing. In one embodiment, the KRAS G12C inhibitor is sotorasib, or a pharmaceutically acceptable salt thereof. In one embodiment, the KRAS G12C inhibitor is adagrasib, or a pharmaceutically acceptable salt thereof.

[0031] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the subject has a TP 53 mutation. The human TP53 gene is located on chromosome 17p, and consists of 11 exons and 10 introns. The wild type p53 protein consists of 393 amino acid residues. Several p53 mutations have been identified in colorectal cancer. Examples of p53 mutations include those described in Li et al., World J Gastroenterol (2015) 21(l):84-93 and Bouaoun et al., Hum Mutat. (2016) 7(9): 865-876. In some embodiments, the TP53 mutation can be selected from C176F, R248W, R248Q, G262V, Y220N, P98fs, E285K, L130H, Q192 and a combination thereof.

[0032] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the subject does not have a TP 53 mutation. In some embodiments, the subject does not have a TP 53 mutation, and has &PIK3CA mutation.

[0033] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the subject has received at least one (c. ., one, two or three) prior systemic therapy. Such first-line standard therapies include immunotherapy, such as an immune checkpoint inhibitor, alone or in combination with chemotherapy, such as platinum-based chemotherapy. Non-limiting examples of platinumbased chemotherapy include cisplatin, carboplatin and oxaliplatin (including pharmaceutically acceptable salts of any of the foregoing).

[0034] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination ofCompound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, can be used to treat a subject that has received prior KRAS G12C inhibitor monotherapy, prior KRAS G12C combination therapy or a prior combination of KRAS G12C inhibitor monotherapy and / or KRAS G12C combination therapy.

[0035] 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, spartalizumab, tebotelimab and volrustomig (including pharmaceutically acceptable salts of any of the foregoing).

[0036] 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).

[0037] 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).

[0038] 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).

[0039] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the subject has received prior KRAS G12C monotherapy or prior KRAS G12C combination therapy (z.e., KRAS G12C inhibitor in combination with a second therapeutic agent, or a pharmaceutically 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, S0S1 inhibitors, etc ).

[0040] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the cancer 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 of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the cancer is a KRAS G12C inhibitor-resistant cancer or a KRAS G12C inhibitor-refractory cancer. In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable saltthereof, provided herein, the cancer is a “KRAS 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, S0S1 inhibitors, etc., (including pharmaceutically acceptable salts thereof)).

[0042] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the cancer 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 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 is inherently resistant.

[0043] In some embodiments of the uses of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the cancer is colorectal cancer, including, but not limited to, metastatic colorectal cancer, advanced colorectal cancer, KRAS G12C-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 of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination ofCompound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the cancer is pancreatic cancer, including, but not limited to, metastatic pancreatic cancer, advanced pancreatic cancer, KRAS G12C-mutated pancreatic 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 of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein, the cancer is non-small cell lung cancer, including, but not limited to, metastatic non-small cell lung cancer, advanced pancreatic cancer, KRAS G12C-mutated non-small cell lung cancer, KRAS G12C inhibitorsensitive 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] Several cancers that can be treated with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, are provided herein. In some embodiments, a cancer can be inherently resistant. In some embodiments, a cancer can acquire resistance.

[0047] When the treatment is a combination of compounds, the order of administration or use of the combination of Compound (A) and KRAS G12C inhibitor (including pharmaceutically acceptable salts thereof) described herein can vary. In some embodiments, Compound (A), or a pharmaceutically acceptable salt thereof, and the KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, are administered or used sequentially. In some embodiments, Compound (A), or a pharmaceutically acceptable salt thereof, can be administered prior to the KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof. In other embodiments, Compound (A), or a pharmaceutically acceptable salt thereof, can be administered subsequent to or after the KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof. In still other embodiments, Compound (A), or a pharmaceutically acceptable salt thereof, can be administered concomitantly or concurrently with the KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof.

[0048] In some embodiments, the use of a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer or methods of treatment with a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, provided herein can decrease the number and / or severity of side effects that can be attributed to monotherapy of the KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof.

[0049] Using a combination of Compound (A), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, 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.

[0050] 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 (z.c., 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.

[0051] 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 the KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, is administered as a monotherapy. For example, the amount of a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, used in a combination described herein can be less compared to the amount of a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, 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 a KRAS G12C inhibitor, or a pharmaceuticallyacceptable salt thereof, 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 toxicides between the compounds of a combination described herein.Pharmaceutical Compositions

[0052] Compound (A), or a pharmaceutically acceptable salt thereof, can be provided in a pharmaceutical composition. Likewise, a KRAS G12C inhibitor, including pharmaceutically acceptable salts thereof, can be provided in a pharmaceutical composition.

[0053] 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.

[0054] 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.

[0055] 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 inj ection, 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.

[0056] 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.

[0057] In some embodiments, a KRAS G12C inhibitor, along with pharmaceutically acceptable salts thereof, can be provided in a pharmaceutical composition that includes Compound (A), or a pharmaceutically acceptable salt thereof. In other embodiments, a KRAS G12C inhibitor, along with pharmaceutically acceptable salts thereof, can be administered in a pharmaceutical composition that is separate from a pharmaceutical composition that includes Compound (A), or a pharmaceutically acceptable salt thereof.

[0058] 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.

[0059] 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.

[0060] 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), or a pharmaceutically acceptable salt thereof, can be administered orally. In some embodiments, Compound (A), or a pharmaceutically acceptable salt thereof can be provided to a subject by the same route of administration as a KRAS G12C inhibitor, along with pharmaceutically acceptable salts thereof. In other embodiments, Compound (A), or a pharmaceutically acceptable salt thereof,can be provided to a subject by a different route of administration as a KRAS G12C inhibitor, along with pharmaceutically acceptable salts thereof.

[0061] 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.

[0062] The compositions may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient. The pack may 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

[0063] As provided herein, in some embodiments, a combination of compounds that includes an effective amount of Compound (A), or a pharmaceutically acceptable salt thereof, and an effective amount of a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, can be used to treat a disease or condition described herein, such as a cancer selected from a colorectal cancer, a pancreatic cancer and a non-small cell lung cancer.

[0064] 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 usedin their normal sense as understood by those skilled in the art. Thus, the cancer can be a recurrent cancer.

[0065] As used herein, a “subj ect” refers to an animal that is the obj ect of treatment, observation or experiment. “Animal” includes cold- and warm-blooded vertebrates and invertebrates such as fish, shellfish, reptiles and, in 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 (z.c., > 1 year and < 18 years old) and / or an infant (i.e., < 1 year old). In other embodiments, the subject can be an adult (z.e., > 18 years old).

[0066] 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.

[0067] 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.

[0068] 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 causedby the cancer, (b) the reduction of tumor size, (c) the elimination of the tumor, and / or (d) longterm disease stabilization (growth arrest) of the tumor.

[0069] 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.

[0070] 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, can be 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 a KRAS G12C inhibitor, or pharmaceutically acceptable salts of the foregoing, 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.

[0071] 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 mostpreferably 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.

[0072] 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.

[0073] 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

[0074] 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. Unless otherwise indicated, all of the cell-line derived xenograft (CDX) and patient-derived xenograft (PDX) models are KRAS G12C-mutated (z.c., contain the KRAS G12C mutation).

[0075] The TP53 gene status (wild-type or mutated) of all CDX and PDX models described in the following examples is provided in Table 1. Table 1 also summarizes the cancer each model represents, whether a model is cell line-derived or patient-derived, whether a model is sensitive or resistant to KRAS G12C inhibitor treatment.Table 1. TP53 Status of CDX and PDX ModelsExample 1 : Combination of Compound (A) and G12C Inhibitor Demonstrates Synergy and Induces DNA Damage and Apoptosis In Vitro in 2D Cellular AssaysIn Vitro 2D Spheroid Growth Assays

[0076] Cell suspensions containing 4 x 103total cells in 100 pL culture media were deposited in each well of a 96-well plate. Plates were incubated at 37 °C with 95% oxygen and 5% CO2 for 24 h to allow cell attachment. After 24 h, Compound (A) (azenosertib, or a pharmaceutically acceptable salt thereof) and a KRAS G12C inhibitor (sotorasib or adagrasib) in DMSO were deposited into the plates using an automated drug dispenser at the indicated concentrations. Total DMSO content was normalized to 0.1% of the total volume in all wells.Plates were incubated at 37 °C with 95% oxygen and 5% CO2 for 168 h. After 168 h, plates were removed from the incubator and allowed to come to room temperature (RT). 100 pL room temperature 2D CTG reagent (Promega, Cat# G7573) was added to each well. Plates were agitated at 520 revolutions per minute (rpm) for 5 mins, allowed to stabilize protected from light for 30 mins, then luminescence was measured on an M5e plate reader (SpectraMax). Percent viability was calculated as percentage of cell viability relative to DMSO-only vehicle control. The percent viability was input into the SynergyFinder tool (http: / / www.synergyfmderplus.org / ) and synergy was output in a heatmap format. The reported synergy score is the Loewe or Bliss synergy score and Synergy Score > 10 is considered synergistic.In Vitro 2D Biomarker Assays

[0077] Cell suspensions were deposited in each well of a 6-well plate. Plates were incubated at 37 °C with 95% oxygen and 5% CO2 for 24 h to allow for cell attachment. After 24 h, Compound (A), a KRAS G12C inhibitor compound (sotorasib or adagrasib) or a combination thereof in DMSO were deposited into the plates using an automated drug dispenser. Plates were incubated at 37 °C with 95% oxygen and 5% CO2 for 24 h. Protein expression was determined for the indicated markers on the JESS Western blot instrument. Quantification of intensity was calculated using ImageJ and represented as fold change compared to DMSO.SiRNA Transfection Assays

[0078] Cells were deposited in a 96-well plate and incubated overnight. Transfection was performed with siControl (Silencer™ Select Negative Control No. 2, #4390847) or siWEEl (Dharmacon, #L-OO5O5O-OO-OOO5) the following day using Lipofectamine RNAiMAX (Thermo Fisher Scientific, #13778150) following the manufacturer’s protocol. Immediately after transfection, compounds were dispensed into the 96-well plate using the Pico 8. After 72 hours, cell viability was assessed using 2D CellTiter- Glo reagent. Transfected cells were collected and lysed on ice in RIPA lysis buffer (Thermo Fisher Scientific, #89901), and protein concentrations were determined using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, #23227). Western blotting was performed using Jess Simple Western System (ProteinSimple) according to the manufacturer’ s standard method for 12-230 EDa Jess separation module (#SM-W004).Cell Cycle Analysis

[0079] Cells were seeded in a 12-well plate, allowed to attach overnight, then treated with the indicated inhibitors for 24 hours. Cells were then pulsed with 10 pM EdU (Click-iT™ Plus EdU Alexa Fluor™ 647, Thermo Fisher, #C 10634) at 37 °C for 2 hours, harvested, washed with 1% BSA in PBS, fixed with Click-iT fixative, and permeabilized with IX Click-iT permeabilization and wash reagent. Click-iT EdU was detected by incubating the cells in Click-iT Plus reaction cocktail at RT for 30 minutes before DNA content staining (FxCycle™ Violet Stain, Thermo Fisher, #F 10347). Samples were analyzed using an Attune™ Flow Cytometer, and data analysis was performed using FlowJo and GraphPad Prism.Biomarker Analysis

[0080] For 2D biomarker analysis, cells were lysed 24 hours after treatment with RIPA buffer (ThermoFisher, #89900), and sonicated on a Bioruptor Pico instrument (Diagenode). All lysis buffers were supplemented with protease and phosphatase inhibitors and quantified by BCA assay. Western blotting was performed using Jess. Chemiluminescence reactions with antibodies were measured and their digital blot images were constructed by the Compass software (Version 6.2). Quantification by densitometry was performed using the area of targeted proteins and normalized to the total protein.

[0081] FIG. 1A depicts synergy heatmaps generated from the SynergyFinder tool in six non-small cell lung cancer (NSCLC) cell lines (NCI-H2122, NCI-H358, NCI-H1792, NCI-H2030 and NCI-H23 that are KRAS G12C inhibitor-sensitive; and SW 1573 that is KRAS G12C inhibitor-resistant). Escalating doses of Compound (A) as a single agent or combined with either sotorasib or adagrasib in escalating doses in a matrixed format were given to the indicated cell lines. The Loewe synergy score was determined for each combination. Synergy Score > 10 is considered synergistic. The data indicates that in all tested cell lines Compound (A) combined with either adagrasib or sotorasib produced synergy scores > 10 in at least one dose combination level, indicating that this combination was synergistic in vitro. Moreover, while Loewe synergy scores are commonly utilized to indicate synergy, the Bliss independence model can also be utilized for combination studies where each compound acts on independent signaling pathways. Bliss synergy scores for each combination as evaluated in FIG. IBdemonstrated similar patterns of synergy to Loewe scores (FIG. 1A), indicating this synergy phenotype is independent of statistical methodology.

[0082] To understand the mechanism of this observed synergy, pharmacodynamic (PD) biomarker changes in response to Compound (A) and KRAS G12C inhibitor monotherapy, and combination treatment in the NCI-H2122 NSCLC cell line were analyzed (FIG. 1C). WEE1 phosphorylates CDK1 at tyrosine 15 (p-CDKl Y15) and loss of this phosphorylation signal is a measure of target engagement by Compound (A). It was found that p-CDKl Y15 was reduced in response to Compound (A), demonstrating suppression of WEE1 kinase activity. Surprisingly, treatment with either sotorasib or adagrasib alone also reduced p-CDKl Y15, possibly due to an extended G1 phase as reported previously. Moreover, combination treatment further reduced p-CDKl Y15 levels, demonstrating additional loss of G2 / M and Gl / S cell cycle checkpoint control. It was found that the downstream KRAS pathway targets p-ERKl / 2 T202 / Y204 and p-S6 S240 / 244 were both reduced in response to treatment with KRAS G12Cinhibitor monotherapy, as well as the combination with Compound (A), demonstrating on-target inhibition of KRAS signaling. Furthermore, a modest increase in levels of H2AX at S139 (yH2AX), a DNA damage marker and proxy for replication stress (RS), was observed in response to either monotherapy, indicating that both inhibitors can individually upregulate some level of DNA damage and RS. Critically, combination treatment resulted in a much stronger upregulation of yH2AX and apoptosis as shown by cleaved caspase 3 (c-Casp-3) and cleaved caspase 7 (c-Casp-7) (FIG. 1C). Cell cycle analysis by DNA content revealed increased G2 / M phase arrest in response to Compound (A) monotherapy treatment, and this effect was exacerbated in the combination. Increased DNA replication fork stalling was also observed (as marked by EdU- DNA staining) with Compound (A) monotherapy or combination treatment, consistent with WEEl’s role in replication fork stability (FIG. ID).

[0083] To determine if PD changes were cell line-specific, the analysis of yH2AX and c-Casp-3 / 7 was expanded to the entire panel of NSCLC cell lines. Consistent with what was observed in NCI-H2122 cells, mild to moderate upregulation of yH2AX and c-Casp-3 / 7 in response to either monotherapy treatment was detected in most cell lines. Combination treatment resulted in markedly higher yH2AX and c-Casp-3 / 7 signals across the majority of models tested, confirming the synergistic effect of Compound (A) with KRAS G12C inhibitors to drive DNA damage, RS and apoptosis (FIG. IE). The magnitude of increases in yH2AXand c-Casp-3 / 7 varied across different cell lines underscoring the heterogeneity of response to both monotherapy and combination treatment. However in 22 of 24 (92%) conditions measured, the magnitude of PD changes in response to combination treatment was greater than monotherapy.

[0084] Evaluation of whether the synergistic effect of Compound (A) and KRAS G12C inhibitors also applies to indications beyond NSCLC was undertaken. Consistent with the observations in NSCLC, matrixed combination treatment with Compound (A) and KRAS G12C inhibitors resulted in synergy in all tested colorectal cancer (CRC) (SW837, SW1463) and pancreatic ductal adenocarcinoma (PDAC) cell lines (MIA-Paca-2) (FIG. IF). Similarly, downstream biomarker changes in target engagement, DNA damage and apoptosis were consistent with those observed in NSCLC cell lines (FIG. 1G), indicating that the mechanism was applicable across NSCLC, CRC and PDAC.

[0085] It was further confirmed that the synergistic cell growth inhibition by the combination of Compound (A) and a KRAS G12C inhibitor is due to the specific targeting of WEE1 by Compound (A) as knockdown of WEE1 by siRNA combined with KRAS G12C inhibition resulted in increased synergy when compared against siControl (FIGS. 1H and II).Example 2: Combination of Compound (A) and G12C Inhibitor Reduces Tumor Cell Growth and Induces DNA Damage and Apoptosis In Vitro in 3D Cellular AssaysIn Vitro 3D Spheroid Growth Assays

[0086] Cell suspensions containing 1.5 x 103total cells in 100 pL 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), a KRAS G12C inhibitor compound (sotorasib or adagrasib) or a combination thereof in DMSO were deposited into the 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. Plates were incubated at 37 °C with 95% oxygen and 5% CO2 for 168 h. Images were taken every 8 hours in an Incucyte instrument and spheroid area was calculated. In Vitro 3D Biomarker Assays

[0087] Cell suspensions containing 1.5 x 103total cells in 100 pL 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 120 h to allow for spheroid formation. After 120 h,Compound (A), a KRAS G12C inhibitor compound (sotorasib or adagrasib) or a combination thereof in DMSO were deposited into the plates using an automated drug dispenser. 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 24 h. Protein expression was determined for the indicated markers on the JESS Western blot instrument. yH2AX and cleaved caspase 3 or cleaved caspase 7 were measured by JESS and quantified using ImageJ.

[0088] It has been demonstrated that RAS-driven models grown as 3D spheroids may more accurately reflect the sensitivity to KRAS G12C inhibition in vivo. To this end, it was sought to validate the 2D synergy findings in a subset of cell lines that could be cultured as anchorage-independent spheroids in ultra-low attachment (ULA) conditions. It was found that combination treatment of NCI-H358, NCI-H1792 and NCLH23 spheroids displayed synergy in 3D conditions, confirming the synergistic effect of that was detected in 2D cultures (FIG. 2A).

[0089] To directly visualize how these 3D spheroid models responded to treatment, live cell imaging-based spheroid growth assays was performed (FIGS. 2B and 2C). When compared to the DMSO control, either monotherapy resulted in statistically significant (p < 0.0001) reduction in spheroid size. Additionally, combination treatment resulted in even greater reduction of spheroid size when compared against either monotherapy (p < 0.0001). These data provide a functional demonstration of tumor cell growth inhibition in anchorageindependent conditions in response to combination treatment.

[0090] Biomarker changes in the spheroid growth environment in response to treatment were evaluated. Similar to 2D conditions, it was found that NCI-H23 spheroids treated with Compound (A) monotherapy resulted in downregulation of p-CDKl Y15 and upregulation of vI I2AX and c-Casp-3 / 7. Similarly, it was found that monotherapy treatment with either sotorasib or adagrasib resulted in downregulation of p-CDKl Y15, p-ERKl / 2 and p-S6, and combination treatment resulted in marked increases in yH2AX and c-Casp-3 / 7, and further downregulation of p-CDKl Y15 relative to monotherapy (FIG. 2D).

[0091] These data further demonstrate the in vitro synergistic activity of WEE1 andKRASG12Cinhibition through increased DNA damage, RS and apoptosis in orthogonal and more physiologically relevant model systems.Example 3: Treatment of an Non-Small Cell Lung Cancer (NSCLC) Cell-Line Xenograft (CDX) Model with Compound (A) and Sotorasib Results in Tumor Regressions and Increased DNA Damage and Apoptosis In VivoCDXModels

[0092] 6-8 weeks old BALB / c nude (MiaPaca-2 that is KRAS G12C-sensitive pancreatic cancer), NOD / SCID (SW837 that is KRAS G12C-sensitive colorectal cancer, NCI- H2122, NCI-H2030, NCI-H1792, SW1573) or NCG (SW1463 that is KRAS G12C-sensitive colorectal cancer) mice were inoculated subcutaneously on the right flank with the single cell suspension of 95% viable tumor cells. PDXModels

[0093] 7-9 weeks old BALB / c nude or athymic nude (Crl:NU(NCr)-Foxnlnu) mice were inoculated subcutaneously in the right flank with tumor fragments from stock mice (2-3 mm in diameter) for tumor development.

[0094] Grouping and treatments started when the mean tumor volume reached about 200 mm3. Vehicle animals were treated daily with 10 mL / kg 20% HP-|3-CD p.o. (oral gavage). Compound (A) was prepared daily in 20% HP- -CD and dosed daily on a 5:2 dosing schedule p.o. at the indicated doses. Sotorasib was prepared daily in 2% HPMC + 1% Tween80 in water and dosed daily p.o. at the indicated doses. Adagrasib was prepared daily in 10% Captisol in 50 mM Citrate Buffer pH = 5.0 and dosed daily p.o. at the indicated doses. Body weight and tumor volume of all animals was measured twice weekly. The measurement of tumor size was performed with a caliper and the tumor volume (mm3) was estimated using the formula: TV = a - b2 / 2 throughout the study, where “a” and “b” are long and short diameters of a tumor, respectively. Animals were euthanized when their individual tumor burden exceeded 2000 mm3or was in a continuing deteriorating condition or close to a comatose state. Mouse PD Studies

[0095] Tumors were harvested 4 hours or 24 hours post single dose. Tumor tissue was snap-frozen in liquid nitrogen or formalin-fixed and embedded in paraffin (FFPE) for further analysis. Ki67 IHC was formed on 5 pm sections. Antigen retrieval was performed with citrate buffer pH 6.0 and blocked with goat serum. Primary rabbit polyclonal Ki67 was stained for 1 hour and detected with goat anti-rabbit-HRP secondary. IHC H-scoring and H&E interpretation were performed by a board-certified veterinary pathologist with experience in laboratory animals and toxicologic pathology. Images had 3 areas counted for % positive cellsin each of the three 40x magnification fields. For biomarker analysis of mouse tissue samples, snap frozen tumor tissues were lysed in RIPA buffer using a tissue homogenizer (BeadBlaster 24R, Benchmark Scientific) and analyzed using Jess.Statistical Analysis

[0096] Unless stated otherwise, all data were statistically analyzed using the GraphPad Prism software (Version 10). Grouped data were analyzed by two-way RM ANOVA. Data are presented as mean ± SEM. Results were considered statistically significant if P < 0.05 (*), P < 0.01 (**), P < 0.001 (***).

[0097] Mice bearing NCI-H2122 cell line-derived xenografts (CDX) were treated with clinically relevant dosing regimens. Monotherapy treatment with Compound (A) once daily for 5 days on and 2 days off (QD 5:2) or sotorasib once daily (QD) resulted in moderate but statistically significant TGI (45% and 77%, respectively; p < 0.0001 vs. vehicle; FIG. 3A). Combination treatment resulted in 104% TGI and led to tumor regressions in all 9 animals (p < 0.0001 vs. either monotherapy; FIGS. 3A and 3B). Further, Compound (A) and sotorasib were well-tolerated over the course of the study as illustrated by minimal changes in mean body weight relative to that of the vehicle group (FIG. 3C).

[0098] To assess PD modulation in vivo, a separate PD study in NCI-H2122 was performed and tumors from animals treated with a single dose of both compounds were analyzed. Similar to the in vitro findings, p-CDKl Y15 levels were decreased in response to either monotherapy and the decrease was more profound when the treatments were combined (FIG. 3D). In contrast to in vitro findings, some tumors displayed rapid reactivation of p- ERK1 / 2 and p-S6 in response to treatment, consistent with reported data. While p-S6 appeared to be inhibited by Compound (A) in the NCI-H2122 xenograft model, this effect was not observed in the in vitro analyses of NCI-H2122 (FIG. 1C), MIA PaCa-2, SW837 (FIG. 1G), or NCI-H23 (FIG. 2D), suggesting this may be a xenograft-specific phenotype. Likewise, the same p-S6 decrease was observed in Compound (A)-treated SW1573 xenografts in vivo (FIG. 3F), but not when the cell line was treated in vitro (FIG. 3E), further corroborating this is likely a phenomenon in xenografts rather than an indication of non-specific kinase inhibition by Compound (A). yH2AX and c-Casp-3 / 7 were only slightly upregulated in response to the monotherapies, but highly increased in response to combination treatment (FIG. 3D). Further tumor analysis demonstrates significantly reduced proliferation by Ki67 IHC and increasedcell death in response to combination treatment relative to either monotherapy (FIGS. 3G-3J). FIG. 31 shows representative images of H&E staining performed on NCI-H2122 tumors treated with one dose of the indicated compounds or the combination thereof, whereby in the upper left image, pink areas in the lx image are large areas of cell death; smaller areas of cell death are in the 20x panel (circled). The 40x panel has a sheet of tumor cells with a prominent mitotic figure (small circle). In the upper right image, the 20x panel has an area of cell death (circle). In the lower left image, the 20x panel has numerous individual dead cells and small clusters (circle), and the 40x panel shows the more epithelial / gland-like tumor cell arrangement. In the lower right image, the 20x and 40x panels have larger foci of dead tumor cells (circles).

[0099] The data demonstrates that although treatment with Compound (A) or sotorasib are moderately efficacious on their own, the combination enhances efficacy and can lead to tumor regression in the NCI-H2122 model.Example 4: Combination of Compound (A) and G12C Inhibitor Improved Efficacy and Drives Tumor Regression in Models of Colorectal Cancer and Pancreatic Cancer

[0100] To determine if in vitro efficacy across tumor types was consistent in vivo, the combination activity in CDX models of CRC (SW837 and SW 1463) or PDAC (MIA PaCa- 2) were evaluated. As observed in NCI-H2122, monotherapy treatment resulted in moderate, but statistically significant (p < 0.0001) TGI when compared to vehicle. Combination treatment with either sotorasib or adagrasib resulted in deeper average TGI (99% and 100% in SW837, 94% and 99% in SW1463, 118% and 107% in MIA PaCa-2, respectively; p < 0.0001) when compared to either monotherapy in all three models (FIGS. 4D-4F). Furthermore, individual animals experienced tumor regression in combination arms in SW837 (sotorasib 3 / 8, adagrasib 5 / 8), SW1463 (sotorasib 1 / 8, adagrasib 4 / 8) and MIA PaCa-2 (sotorasib 8 / 8, adagrasib 7 / 8; FIGS. 4A-C). Additionally, 3 / 8 (38%) of the animals in the combination arm of the MIA PaCa-2 study experienced complete tumor regression (FIG. 4C). This combination was well -tolerated across all tested models (FIGS. 4G-4I).

[0101] The data demonstrates that the combination of Compound (A) with KRAS G12C inhibitors drives significantly increased TGI relative to monotherapy and high rates of regression in tumor models of CRC and PDAC. Only minor differences were observedbetween sotorasib and adagrasib, indicating that inhibition of KRAS and its downstream activity are the mechanistic basis for synergy with Compound (A).Example 5: Combination of Compound (A) and G12C Inhibitor Increases the Depth and Duration of Tumor Responses In Vivo

[0102] Durable responses to therapy are equally important and remain a critical unmet need in patients with KRAS G12C mutant cancers. To assess whether the addition of Compound (A) extends the duration of response to KRAS G12C inhibitors, the combination in the NCI-H2030 and NCI-H1792 NSCLC CDX models were evaluated. In response to Compound (A) monotherapy, the NCI-H2030 CDX model exhibited moderate, dosedependent TGI that was statistically significant relative to vehicle (p < 0.0001). Sotorasib monotherapy displayed early regression, but rapid recurrence and progression within 27 days. Surprisingly, the combination of sotorasib and Compound (A) not only increased depth of response, but extended the time until progression by at least 2.5-fold (FIG. 5A) and further resulted in a significant increase in median survival (p = 0.007, FIG. 5D). Additionally, the combination of Compound (A) and sotorasib continued to be well-tolerated throughout the extended treatment period of 67 days (FIG. 4J).

[0103] In the NCI-H1792 NSCLC model, Compound (A) monotherapy resulted in moderate, but statistically significant TGI relative to vehicle (p < 0.0001). Although adagrasib alone resulted in durable regression for over 40 days, tumors rapidly regrew in the low dose adagrasib (50 mg / kg QD) group when treatment was withdrawn (FIG. 5B). In contrast, tumor regression was maintained in 7 of 9 (78%) animals in the corresponding combination group of Compound (A) and adagrasib (FIG. 5C). Additionally, high dose adagrasib (100 mg / kg QD) extended the duration of regression longer than the lower dose, yet tumor regrowth was eventually observed in all animals by the end of the study (FIG. 5B). Combination treatment at the higher dose again maintained near complete regression in 5 of 6 animals (83%) at the end of the study (FIG. 5C). Furthermore, both combination arms significantly extended median survival over monotherapy treatment at either adagrasib dose (FIGS. 5E and 5F). Moreover, the combination was well-tolerated at all doses during the treatment period as illustrated by minimal body weight loss (FIG. 4K).

[0104] The data establishes that the combination of Compound (A) with KRAS G12C inhibition can extend the depth and duration of response in NSCLC models which arealready relatively sensitive to KRAS G12C inhibitors, effectively delaying the onset of resistance.Example 6: Combination of Compound (A) and G12C Inhibitor Overcomes Innate Resistance to G12C Inhibitors In Vivo

[0105] Many patients respond to KRAS G12C inhibition, however, a substantial proportion of patients are inherently refractory. Combination activity in models that are inherently resistant (< 60% TGI) KRAS G12C inhibition was evaluated. The SW1573 model displayed resistance to sotorasib and adagrasib in vitro (FIG. 1A) and resulted in only 60% TGI in response to 100 mg / kg adagrasib (FIG. 6A). This model is also resistant to Compound (A), as the monotherapy dose of 60 mg / kg QD 5:2 resulted in a relatively weak response of 31% TGI. Surprisingly, combination treatment was significantly more efficacious than either monotherapy (p < 0.0001) resulting in 84% TGI and was well tolerated (FIG. 4L).

[0106] To test this combination in a clinically relevant model of intrinsic resistance, the CR2528 PDX model of CRC was used. CR2528 exhibited resistance to both Compound (A) and adagrasib monotherapy at high doses, with a TGI of 23% and 1% respectively. Even in this highly resistant model, the combination led to approximately 3-fold increased TGI (76%, p < 0.0001) compared with Compound (A) monotherapy (FIG. 6B) and was well- tolerated (FIG. 4M). This data demonstrates that tumor models bearing some level of intrinsic resistance to Compound (A) or KRAS G12C inhibitor monotherapy can benefit from the combination treatment.

[0107] To determine if the combination can drive TGI in a model with acquired resistance to KRAS G12C inhibition, sotorasib-resistant NCI-H2030 xenografts that were derived through continuous exposure to sotorasib in vivo (hereafter referred to as NCI-H2030- R) were utilized. Treatment in the NCI-H2030-R model was tolerated (FIG. 4N) and displayed high levels of resistance to sotorasib monotherapy (FIG. 6C), while maintaining sensitivity to Compound (A) similar to that of the NCI-H2030 parental line (FIG. 5A). Combination treatment resulted in a 20% increase in TGI (82% vs. 62%) with one animal experiencing tumor regression, although statistical significance was not achieved (p=0.07).

[0108] To further assess how widely applicable this combination is in clinically translatable models, a mouse PDX screen in 14 NSCLC or CRC PDX models with varying responses to KRAS G12C inhibition was conducted, including paired sensitive and resistantmodels derived via continuous exposure in vivo. Ten out of 14 (71%) models derived additional benefit from combination therapy over sotorasib monotherapy, including statistically significant differences in the ST6365, and ST1989 models (p < 0.05) (FIG. 6D). Furthermore, it was observed that the combination was beneficial in two paired resistant models irrespective of relative sensitivity to sotorasib monotherapy.

[0109] Altogether, the data shows that the combination of Compound (A) and KRAS G12C inhibition is generally superior to monotherapy across a range of models with intrinsic and acquired resistance to KRAS G12C inhibitors, both of which represent unmet clinical needs.

[0110] 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), or a pharmaceutically acceptable salt thereof, and a KRAS G12C inhibitor, or a pharmaceutically acceptable salt thereof, for treating a cancer in a subject, wherein:Compound (A)or a pharmaceutically acceptable salt thereof; and the cancer is a KRAS G12C inhibitor-resistant cancer or a KRAS G12C inhibitorrefractory cancer and is selected from colorectal cancer, pancreatic cancer and non-small cell lung cancer.

2. The use of claim 1, wherein the subject has a KRAS G12C mutation.

3. The use of claim 2, wherein the subject has a TP53 mutation.

4. The use of claim 3, wherein the TP53 mutation is selected from the group consisting of C176F, R248W, R248Q, G262V, Y220N, P98fs, E285K, L130H, Q192 and a combination thereof.

5. The use of claim 2, wherein the subject does not have a TP 53 mutation.

6. The use of claim 5, wherein the subject has a PIK3CA mutation.

7. The use of any one of claims 1 to 6, wherein the KRAS G12C inhibitor is selected from the group consisting of sotorasib, adagrasib, garsorasib, divarasib, GF-105, JDQ443, LY-3537982, GH-35, glecirasib, FMC-376, HS-10370, JMKX-001899, JS-116, YL- 15293, ZG-19018, BEBT-607, BI-1823911, BPI-421286, D3S-001, ERAS-3490, GEC-255, MK-1084, RG-6330, RMC-6291, TEB-17231, BBO-8520, LF0001, TSN333, ABSK-071, ADGN-121, ADGN-122, ADGN-123, ADGN-531, AFNT-212, APG-1842, ARS-1620, ARS- 853, ASP-2453, AST-NS1902, AU-10458, AU-8653, AZD-4625, AZD-4747, BPI-2361, EB- 160, EB-TM1, GRAD-1405, ICP-915, K20, KP-14, LC-2, MRTX-1257, R023, RM-007, RM- 018, RM-032, UCT-00104, VRTX-126, WDB-178, XNW-14011, YF135, BI-2493, BI-2865 and HYP-2A, or a pharmaceutically acceptable salt of any of the foregoing.

8. The use of claim 7, wherein the KRAS G12C inhibitor is sotorasib, or a pharmaceutically acceptable salt thereof.

9. The use of claim 7, wherein the KRAS G12C inhibitor is adagrasib, or a pharmaceutically acceptable salt thereof.

10. The use of any one of claims 1 to 9, wherein the subject has received at least one prior systemic therapy.

11. The use of claim 10, wherein the at least one prior systemic therapy comprises immunotherapy, platinum-based chemotherapy or a combination thereof.

12. The use of claim 10 or 11, wherein the subject has further received prior KRAS G12C inhibitor monotherapy, prior KRAS G12C combination therapy or a prior combination of KRAS G12C inhibitor monotherapy and KRAS G12C combination therapy.

13. The use of any one of claims 1 to 12, wherein the cancer is colorectal cancer.

14. The use of any one of claims 1 to 12, wherein the cancer is pancreatic cancer.

15. The use of any one of claims 1 to 12, wherein the cancer is non-small cell lung cancer.

16. The use of any one of claims 1 to 15, wherein the cancer is a KRAS G12C inhibitor-sensitive cancer.

17. The use of any one of claims 1 to 15, wherein the cancer is a KRAS G12C inhibitor-resistant cancer or a KRAS G12C inhibitor-refractory cancer.

18. The use of any one of claims 1 to 15 or 17, wherein the cancer is a KRAS G12C inhibitor-resistant cancer.

19. The use of any one of claims 1 to 15 or 17, wherein the cancer is a KRAS G12C inhibitor-refractory cancer.

20. The use of any one of claims 1 to 15 or 17, wherein the cancer is inherently resistant or the cancer acquires resistance.

21. The use of claim 20, wherein the cancer is inherently resistant.

22. The use of claim 20, wherein the cancer acquires resistance.

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